ArticleLiterature Review

Immunologic Interactions of T Lymphocytes with Vascular Endothelium

February 1991
Advances in Immunology 50:261-302

February 1991
50:261-302

Source
PubMed

Authors:

The data presented in this review establish that cultured human endothelial cells have the capacity to present antigens to T cells and to do so in the context of costimulators that lead to effective T cell activation. These activities raise the possibility that venular ECs, at sites of delayed hypersensitivity reactions, could be the primary antigen-presenting cell to circulating memory T cells. This putative role of ECs can explain the rapid rate of initiation of memory responses because ECs are uniquely positioned to have physical access to the pool of circulating memory T cells. Studies also suggest that ECs may present alloantigens to circulating T cells in the context of transplantation, thereby initiating rejection reactions. Nevertheless, we repeat our caveat that these proposed antigen-presenting functions of ECs have not been established in vivo. Cytokine-mediated changes, particularly induction of adhesion molecules and synthesis of lymphocyte-activating cytokines, such as IL-8, provide ECs with the potential to recruit memory T cells to inflammatory sites independent of antigen specificity. Although these functions have also not been rigorously shown to occur in vivo, immunocytochemical studies of experimental and pathological tissues provide significant support for this proposal. Similar adhesive and activating functions of ECs may apply to preferential homing of pre-T cells to thymus and naive T cells to lymph node. We conclude by noting that the weight of evidence reviewed here supports the proposal that the vascular endothelium be considered an integral part of the in vivo immune system.

Cellular interactions between lymphocytes and retinal endothelial cells

Thesis

Jan 1994

Yufei Wang

Cellular interactions between lymphocytes and endothelial cells derived from retinal and brain microvasculatures are believed to play an important part in the pathogenesis of inflammatory diseases of the central nervous system. In this in vitro study we have investigated these interactions in two aspects; (1) lymphocyte adhesion to and migration across retinal endothelial cells, and (2) antigen presenting properties of retinal and brain endothelial cells. Under normal conditions the level of lymphocyte adhesion to retinal endothelial cells was around 5 %, but this low level of adhesion could be significantly upregulated following endothelial treatment with the cytokines, IFN-γ, IL-1 and IL-4, astrocyte conditioned medium and forskolin. Lymphocyte activation with Con A also increased the adhesion which could be further augmented by activating the endothelial cells with the cytokines, astrocyte conditioned medium and forskolin. Although CD8+ T cells in normal and Con A activated lymphocyte populations adhered to endothelial cells more efficiently than CD4+ T cells, the antigen-specific activated CD4+ T-cell lines (S-Ag specific T cell lines) exhibited the greatest degree of adhesion and were also capable of migrating across retinal endothelial monolayers. The role of adhesion molecules in lymphocyte adhesion and migration has been examined. Lymphocytes express both LFA-1 and VLA-4 and activation of lymphocytes with Con A and S-Ag can increase the expression of LFA-1, but not VLA-4. Resting cultured retinal endothelial cells express ICAM-1 and its expression was significantly increased by IFN-γ and IL-1. Treatment of lymphocytes with the monoclonal antibodies to LFA-1 and VLA-4 significantly inhibited the adhesion of Con A activated lymphocytes and S-Ag specific T cell lines to both resting and IL-1 activated retinal endothelial cells. Pre-incubation of retinal endothelial cells with the monoclonal antibody to ICAM-1 only showed an inhibitive effect on the adhesion of S-Ag specific T cell lines, but not Con A activated lymphocytes. Furthermore, the antibodies to LFA-1 and ICAM-1 also blocked the migration of S-Ag specific T cell lines across both resting and IL-1 activated endothelial monolayers and the antibody to VLA-4 only inhibited the migration across IL-1 activated endothelial monolayers. Cultured retinal and brain endothelial cells are capable of expressing MHC class II molecules following treatment of IFN-γ. With both sets of endothelial cells, only class II I-A, but not I-E molecules, could be induced significantly by day 3 with brain EC expressing lower levels of I-A. IFN-γ also caused a concomitant increase in the level of MHC class I molecules. In comparison with expression of adhesion molecules, induced expression of MHC class II molecules appeared to be slower than enhanced expression of ICAM-1 which occurred within 18 hours following treatment with IFN-γ. Retinal and brain EC were capable of presenting S-antigen to S-antigen specific CD4+ T-cell lines resulting in both T-cell proliferation and cytotoxicity. In contrast to the cytotoxic effect which was demonstrated with confluent endothelial monolayers, significant T-cell proliferation was only seen when subconfluent, rather than confluent endothelial cells were used as the antigen presenting cells. Optimal T-cell proliferation was observed at the ratio of T cells to the endothelial cells in 2:5. Subconfluent endothelial cells also resulted in a greater IL-2 production than confluent cells. Although retinal endothelial cells were capable of producing TGF-B, removal of this factor with the anti-TGF-B monoclonal antibody from the subconfluent proliferation assay attenuated the T-cell response. Contrary to these findings, the addition of exogenous TGF-B to the media in the antigen presentation assays with either confluent or subconfluent endothelial cells did not affect the degree of T-cell proliferation and IL-2 production. Despite demonstrating significant T cell responses to retinal and brain endothelial cell antigen presentation, they remained less efficient than professional antigen presenting cells such as thymocytes.

CD86 (B70/B7–2) on endothelial cells co-stimulates allogeneic CD4 + T cells

Article

Aug 1995

In vascularized organ transplantation, vascular endothelial cells (EC) confronting recipient T cells are potentially significant APC initiating cellular immune responses that lead to rejection. In the present study, we studied the ability of human EC to stimulate allogeneic T cells and the co-stimulatory molecules involved in this response. On both human umbilical vein endothelial cells (HUVEC) and microvascular endothelial cells (MVEC), MHC class I, intercellular adhesion molecule (ICAM)-1 and CD86 were constitutively expressed as assessed by flow cytometry. After IFN-γ treatment, MHC class II expression was induced, and MHC class I and ICAM-1 were up-regulated. In contrast, the expression of CD86 was unchanged and CD80 was undetectable even after IFN-γ treatment Highly purified CD4+ T cells proliferated in response to IFN-γ-treated allogeneic HUVEC and MVEC, and this response was efficiently blocked by mAb to MHC class II, ICAM-1 and CD86. Furthermore, the addition of anti-CD86 mAb to the primary culture with allogeneic EC resulted in the induction of alloantigen-specific anergy. These results suggest that CD86 expressed on EC plays a critical role in initiating cellular immune responses to vascularized allografts and would be an important target for immune intervention.

Biomarkers of Endothelial Cell Activation Serve as Potential Surrogate Markers for Drug-induced Vascular Injury

Article

Full-text available

Oct 2010
TOXICOL PATHOL

Drug-induced vascular injury (DIVI) is a nonclinical finding that often confounds the toxicological evaluation of investigational drugs, but there is an absence of qualified biomarkers that can be used to detect and monitor its appearance in animals and patients during drug development and clinical use. It is well known that endothelial cell (EC) activation plays a key role in the expression and evolution of DIVI, and the various immunological and inflammatory factors involved in its expression may serve as potential biomarker candidates. Activated ECs change their morphology and gene expression, generating endothelial adhesion molecules, pro-coagulant molecules, cytokines, chemokines, vasodilators, nitric oxide, and acute-phase reactants. This review provides a brief historical background of EC activation and the search for biomarkers of early EC activation for monitoring DIVI. At present, no biomarkers of EC activation have been qualified to predict DIVI in the nonclinical or clinical context, and a robust pathologic foundation for their use is still lacking. We propose three categories of EC activation biomarkers: recommended surrogate markers, potentially useful markers, and emerging candidate markers. This review alerts pharmaceutical companies, research institutions, and regulatory agencies to the continuing need for reliable biomarkers of EC activation in drug development.

Functional interactions of T cells with endothelial cells: The role of CD40L-CD40-mediated signals

Article

Dec 1995

Michael Yellin

CD40 is expressed on a variety of cells, including B cells, monocytes, dendritic cells, and fibroblasts. CD40 interacts with CD40L, a 30-33-kD activation-induced CD4+ T cell surface molecule. CD40L-CD40 interactions are known to play key roles in B cell activation and differentiation in vitro and in vivo. We now report that normal human endothelial cells also express CD40 in situ, and CD40L-CD40 interactions induce endothelial cell activation in vitro. Frozen sections from normal spleen, thyroid, skin, muscle, kidney, lung, or umbilical cord were studied for CD40 expression by immunohistochemistry. Endothelial cells from all tissues studied express CD40 in situ. Moreover, human umbilical vein endothelial cells (HUVEC) express CD40 in vitro, and recombinant interferon gamma induces HUVEC CD40 upregulation. CD40 expression on HUVEC is functionally significant because CD40L+ Jurkat T cells or CD40L+ 293 kidney cell transfectants, but not control cells, upregulate HUVEC CD54 (intercellular adhesion molecule-1), CD62E (E-selectin), and CD106 (vascular cell adhesion molecule-1) expression in vitro. Moreover, the kinetics of CD40L-, interleukin 1-, or tumor necrosis factor alpha-induced CD54, CD62E, and CD106 upregulation on HUVEC are similar. Finally, CD40L-CD40 interactions do not induce CD80, CD86, or major histocompatibility complex class II expression on HUVEC in vitro. These results demonstrate that CD40L-CD40 interactions induce endothelial cell activation in vitro. Moreover, they suggest a mechanism by which activated CD4+ T cells may augment inflammatory responses in vivo by upregulating the expression of endothelial cell surface adhesion molecules.

MEK5 is Activated by Shear Stress, Activates ERK5 and Induces KLF4 to Modulate TNF Responses in Human Dermal Microvascular Endothelial Cells

Article

Feb 2011
MICROCIRCULATION

ECs lining arteries respond to LSS by suppressing pro-inflammatory changes, in part through the activation of MEK5, ERK5 and induction of KLF4. We examined if this anti-inflammatory pathway operates in human ECs lining microvessels, the principal site of inflammatory responses. We used immunofluorescence microscopy of human skin to assess ERK5 activation and KLF4 expression in HDMECs in situ. We applied LSS to or overexpressed MEK5/CA in cultured HDMECs and assessed gene expression by microarrays and qRT-PCR and protein expression by Western blotting. We assessed effects of MEK5/CA on TNF responses using qRT-PCR, FACS and measurements of HDMEC monolayer electrical resistance. We used siRNA knockdown to assess the role of ERK5 and KLF4 in these responses. ERK5 phosphorylation and KLF4 expression is observed in HDMECs in situ. LSS activates ERK5 and induces KLF4 in cultured HDMECs. MEK5/CA-transduced HDMECs show activated ERK5 and increased KLF4, thrombomodulin, eNOS, and ICAM-1 expression. MEK5 induction of KLF4 is mediated by ERK5. MEK5/CA-transduced HDMECs are less responsive to TNF, an effect partly mediated by KLF4. MEK5 activation by LSS inhibits inflammatory responses in microvascular ECs, in part through ERK5-dependent induction of KLF4.

Costimulator deficient antigen presentation by an endothelial cell line induces a nonproliferative T cell activation response without anergy

Article

Nov 1993

J. D. St. Louis

The ability of endothelial cells to activate helper T (Th) cells by antigen presentation was studied using the murine endothelial cell line SVEC4-10 and antigen-specific murine T cell clones. SEVEC4-10 cells constitutively express vascular cell adhesion molecule 1 but not intercellular adhesion molecule 1. Interferon gamma (IFN-gamma) treatment of these cells induced class II major histocompatibility complex (MHC) expression and antigen-presenting capabilities, but did not alter surface integrin expression. IFN-gamma-treated SVEC4-10 cells were competent at mediating antigen-dependent cytokine production and proliferation of a Th2 clone. In contrast, endothelial antigen presentation to Th1 cells did not stimulate T cell proliferation. The addition of MHC mismatched spleen cells as a source of costimulatory molecules resulted in the ability of the endothelial cells to stimulate Th1 cell proliferation in an antigen-specific manner. The failure of the endothelial cell line alone to support Th1 cell proliferation correlated with the failure to stimulate interleukin 2 (IL-2) gene expression. T cell exposure to the endothelial cells plus antigen resulted in upregulation of IL-2 receptors and an enhanced response to subsequent antigen presentation by splenic antigen-presenting cells. Despite the lack of functional costimulators for IL-2 expression, antigen presentation by the endothelial cell line did not induce Th1 cell anergy, indicating that costimulator deficiency for IL-2 expression is not obligatorily linked to anergy induction. Thus, endothelial cells are capable of presenting antigens to helper T lymphocytes, but stimulate only partial T cell responses. These partial responses may serve to selectively stimulate transmigration of antigen-specific T cells and may enhance functional responses upon subsequent, extravascular antigen exposure.

Benefits of Reiki in older individuals with chronic pain

Article

Full-text available

Dec 2014

Reiki se caracteriza por ser una terapia complementar llevado a cabo a través de la imposición de manos en el ser humano con la intención de restablecer el equilibrio físico, mental y espiritual. El objetivo de esta investigación fue identificar y analizar los beneficios experimentados con la práctica de Reiki, en sujetos ancianos con dolor crónico no oncológica. Se trata de un estudio cualitativo, descriptivo y exploratorio. Para la recolección se utilizó la entrevista semiestructurada, con cuestiones abiertas y cerradas. Los datos fueron recogidos entre julio y agosto de 2012. Los sujetos del estudio fueron diez ancianos con quejas de dolor crónico no oncológica, presentó las cinco sesiones de Reiki. Evaluación de resultados se considera el análisis del contenido propuesto por Bardin. Se concluyó que esta práctica terapéutica mejora significativamente las quejas de dolor crónico, además de contribuir al equilibrio de las necesidades físicas, mentales, emocionales y espirituales de los ancianos.

Liver and Intestinal Transplantation Liver and Intestinal Transplantation

Article

The Vademecum series includes subjects generally not covered in other handbook series, especially many technology-driven topics that reflect the increasing influence of technology in clinical medicine. The name chosen for this comprehensive medical handbook series is Vademecum, a Latin word that roughly means "to carry along". In the Middle Ages, traveling clerics carried pocket-sized books, excerpts of the carefully transcribed canons, known as Vademecum. In the 19th century a medical publisher in Germany, Samuel Karger, called a series of portable medical books Vademecum. The Landes Bioscience Vademecum books are intended to be used both in the training of physicians and the care of patients, by medical students, medical house staff and practicing physicians. We hope you will find them a valuable resource.

Antigenspezifische Migration von T-Helfer-Zellen im murinen Inflammationsmodell

Article

Running title: CCR5 in P. berghei ANKA infection

Article

Full-text available

Over-expression of ICAM-1, VCAM-1 and ELAM-1 might influence tumor progression in colorectal cancer

Article

Feb 1998
INT J CANCER

Adhesion molecules might play a role in tumor progression. We investigated expression of the adhesion molecules ICAM-1, VCAM-1 and ELAM-1 in 24 primary colorectal carcinomas using immuno-histochemistry and Northern blot analysis. Normal colonic tissue from the same patients served as controls. ICAM-1 immunostaining was restricted to the intercellular matrix and vascular endothelial cells. The vast majority of normal tissue samples revealed only faint ICAM-1 immunoreactivity. However, moderate to strong immunostaining was found in 86% of cancerous sections. The ICAM-1 immunoreaction was more intense in well-differentiated carcinomas as well as in the adenomatous parts and transition zones of cancers. Similarly, the cancers exhibited markedly enhanced VCAM-1 and ELAM-1 immunostaining in the endothelial cells of small blood vessels. The intense vascular immunostaining by ICAM-1 and VCAM-1 was associated with a strong presence of CD3-positive T lymphocytes, whereas ELAM-1 immunoreactivity did not correlate with round cell infiltration. On Northern blot analysis, ICAM-1, VCAM-1 and ELAM-1 mRNA levels were increased in 67%, 57% and 63% of carcinomas, respectively, in comparison with normal tissue samples. Densitometric analysis of Northern blots revealed an increase in ICAM-1 by 2.1-fold, an increase in VCAM-1 by 3.4-fold and an increase in ELAM-1 by 2.2-fold in cancerous tissues compared to normal controls. Over-expression of ICAM-1 might prevent cell–cell disruption and, hence, tumor dissemination. Furthermore, over-expression of ICAM-1 and VCAM-1, but not ELAM-1, might favor host anti-tumor defense by trafficking of lymphocytes. Int. J. Cancer (Pred. Oncol.) 79:76–81, 1998. © 1998 Wiley-Liss, Inc.

Higher body mass index (BMI) is associated with reduced glucocorticoid inhibition of inflammatory cytokine production following acute psychosocial stress in men

Article

Sep 2008
PSYCHONEUROENDOCRINO

Body mass index (BMI) and mental stress seem to exert part of their cardiovascular risk by eliciting inflammation. However, the adverse effects of stress on inflammatory activity with BMI are not fully understood. We investigated whether higher BMI is associated with reduced glucocorticoid inhibition of inflammatory cytokine production following stress in men while controlling for age and blood pressure. We measured glucocorticoid inhibition of lipopolysaccharide (LPS)-stimulated release of the proinflammatory cytokine tumor necrosis factor (TNF)-alpha. Forty-two men (age range 21-65 years; BMI range 21-34 kg/m(2)) underwent the Trier Social Stress Test (combination of mock job interview and mental arithmetic task). Whole blood samples were taken immediately before and after stress, and during recovery up to 60 min post-stress. Glucocorticoid sensitivity of LPS-stimulated TNF-alpha expression was assessed in vitro with and without coincubating increasing doses of dexamethasone. Moreover, salivary cortisol was measured during the experiment and on a normal day for assessment of baseline circadian cortisol. Higher BMI was associated with lower glucocorticoid sensitivity of monocyte TNF-alpha production after stress (main effect of BMI: p<0.001) and with more pronounced decreases of glucocorticoid sensitivity following stress (interaction of stress-by-BMI: p=0.002). Neither LPS-stimulated TNF-alpha release nor baseline glucocorticoid sensitivity were associated with BMI. Similarly, BMI was not associated with salivary cortisol, either in reaction to stress or in circadian cortisol secretion. Our data suggest that with increasing BMI, glucocorticoids are less able to inhibit TNF-alpha production following stress. This might suggest a new mechanism linking BMI with elevated risk for adverse cardiovascular outcomes following stress.

T cell reactions of Eimeria bovis primary- and challenge-infected calves

Article

Full-text available

Feb 2010
Parasitol Res

Eimeria bovis infections commonly have clinical impact only on young animals, as homologous reinfections generally are under immunological control. So far, the nature of the immune responses delivering protection to calves has not been investigated. In this study we therefore analysed local and peripheral proliferative T cell activities of primary and challenge-infected calves and investigated the occurrence of T cell phenotypes in the peripheral blood and in mucosal gut segments isolated either by bioptic means or by necropsies.We show that lymphocytes of E. bovis-infected calves exhibit effective, transient antigen-specific proliferative responses in the course of prepatency of primary infection but fail to react after homologous reinfection suggesting early abrogation of parasite development. Whilst in primary infection an expansion of peripheral CD4+ T cells was observed, reinfection had no effect on the proportions of CD4+, CD8+ subsets or gammadeltaTCR+ T cells. In contrast, both E. bovis primary and challenge infections had an impact on local tissue T cell distribution. Primary infection was characterised by a CD4+ T cell infiltration early in prepatency in ileum and later in colon mucosa, whereas CD8+ T cells were only found accumulating in the latter gut segment. Challenge infection led to infiltration of both CD4+ and CD8+ T cells in small intestine and large intestine segments indicating protective functions of both cell types. In contrast, infiltration of ileum and colon mucosa with gammadeltaTCR+ T cells was restricted to primary infection.

Assoziation lymphozytärer Oberflächenmoleküle mit Blut-Hirn-Schrankenstörungen

Article

Martin Breckner

Untersuchungen zur Rolle des HIV-1-Tat-Proteins in der AIDS-assoziierten Vaskulopathie

Article

Kathrin Matzen

Bei Patienten, die mit dem humanen Immundefizienzvirus-1 (HIV-1) infiziert sind, kommt es häufig zu krankhaften Veränderungen des Endothels, die zu einer Fehlfunktion des Gefäßsystems führen. Klinischer Ausdruck dieser als acquired immune deficiency syndrome (AIDS)-assoziierten Vaskulopathie bezeichneten Veränderungen sind Schädigungen des Aortenendothels, die mit einer erhöhten Adhäsion mononukleärer Zellen an das Endothel einhergehen, Defekte der Blut-Hirn-Schranke, die zur Entstehung von Demenz beitragen, sowie das Kaposi-Sarkom (KS), das durch eine sehr starke Extravasation von T-Zellen und Monozyten gekennzeichnet ist. In dieser Arbeit wird gezeigt, dass das regulatorische HIV-1-Tat-Protein und das inflammatorische Zytokin TNF-a synergistisch die Adhäsion der promonozytären Zelllinie U937 und von PBMZ an humane mikrovaskuläre Endothelzellen (HMVEZ) erhöht. Die adhäsionsfördernde Wirkung wurde selektiv bei HIV-1-Tat beobachtet, andere virale Proteine des HIV-1, wie Negativfaktor (Nef) und das Glykoprotein gp41, hatten keinen Einfluss auf die Adhäsion. Anhand zellspezifischer Marker wurde gezeigt, dass HIV-1-Tat in periphere mononukleäre Blutzellen (PBMZ) spezifisch die Adhäsion von Monozyten und T-Zellen erhöhte, jedoch nicht von B-Zellen. Intravital-mikroskopische Untersuchungen an der Maus bestätigten in vivo, dass HIV-1-Tat und TNF-a synergistisch die Adhäsion von Leukozyten an das Endothel erhöhten. HIV-1-Tat reguliert die Expression einer großen Anzahl zellulärer Gene. Diese Fehlregulation durch HIV-1-Tat könnte an der Enstehung der AIDS-assoziierten Vaskulopathie beteiligt sein. Im zweiten Teil dieser Arbeit wird die parakrine Wirkung von HIV-1-Tat auf die Genexpression in Monozyten mittels der suppressed subtractive hybridization (SSH)-Methode untersucht. Hierbei wurde O-linked N-Acetylglucosamine-transferase (OGT) als Gen identifiziert, dessen Expression durch HIV-1-Tat unterdrückt wird. Bisher ist bekannt, dass OGT ein Repressor der basalen Transkription und der SP-1-regulierten Transkription ist. Die Expression von OGT wurde sowohl auf mRNA-Ebene als auch auf Protein-Ebene durch HIV-1-Tat und VEGF121 gehemmt, wobei die Regulierung über den VEGF-Rezeptor Flt-1 vermittelt wurde. Weitere Faktoren wie inflammatorische Zytokine (TNF-a, IL-1b, IFN-g und IL-2), angiogene Wachstumsfaktoren (bFGF und VEGF165) und Chemokine (IL-8, MIP-1a, IP-10, MCP-1 und SDF-1a) hatten keine hemmende Wirkung auf die OGT-Expression. Die schnelle Abnahme von intrazellulärem OGT-Protein wurde weder durch lysosomale Proteasen noch durch Proteasen des Proteasoms verursacht. Expressionsstudien an PBMZ von fünf verschiedenen Probanden zeigten, dass bei zwei Probanden die OGT-Konzentration durch HIV-1-Tat zunahm, bei zweien nahm sie ab und bei einer Person gab es keine Veränderung. Diese Ergebnisse belegen, dass HIV-1-Tat entscheidend an der Entstehung der AIDS-assoziierten Vaskulopathie, insbesondere von KS, beteiligt sein könnte. Die Repression von OGT durch HIV-1-Tat könnte die weitreichende Wirkung des HIV-1-Tat-Proteins auf zelluläre und virale Gene erklären.

Plasmodium yoelii 17XL infection up-regulates RANTES, CCR1, CCR3 and CCR5 expression, and induces ultrastructural changes in the cerebellum

Article

Full-text available

Jan 2005
MALARIA J

Abstract Background Malaria afflicts 300–500 million people causing over 1 million deaths globally per year. The immunopathogenesis of malaria is mediated partly by co mplex cellular and immunomodulator interactions involving co-regulators such as cytokines and adhesion molecules. However, the role of chemokines and their receptors in malaria immunopathology remains unclear. RANTES (Regulated on Activation Normal T-Cell Expressed and Secreted) is a chemokine involved in the generation of inflammatory infiltrates. Recent studies indicate that the degradation of cell-cell junctions, blood-brain barrier dysfunction, recruitment of leukocytes and Plasmodium -infected erythrocytes into and occlusion of microvessels relevant to malaria pathogenesis are associated with RANTES expression. Additionally, activated lymphocytes, platelets and endothelial cells release large quantities of RANTES, thus suggesting a unique role for RANTES in the generation and maintenance of the malaria-induced inflammatory response. The hypothesis of this study is that RANTES and its corresponding receptors (CCR1, CCR3 and CCR5) modulate malaria immunopathogenesis. A murine malaria model was utilized to evaluate the role of this chemokine and its receptors in malaria. Methods The alterations in immunomodulator gene expression in brains of Plasmodium yoelii 17XL-infected mice was analysed using cDNA microarray screening, followed by a temporal comparison of mRNA and protein expression of RANTES and its corresponding receptors by qRT-PCR and Western blot analysis, respectively. Plasma RANTES levels was determined by ELISA and ultrastructural studies of brain sections from infected and uninfected mice was conducted. Results RANTES (p < 0.002), CCR1 (p < 0.036), CCR3 (p < 0.033), and CCR5 (p < 0.026) mRNA were significantly upregulated at peak parasitaemia and remained high thereafter in the experimental mouse model. RANTES protein in the brain of infected mice was upregulated (p < 0.034) compared with controls. RANTES plasma levels were significantly upregulated; two to three fold in infected mice compared with controls (p < 0.026). Some d istal microvascular endothelium in infected cerebellum appeared degraded, but remained intact in controls. Conclusion The upregulation of RANTES, CCR1, CCR3, and CCR5 mRNA, and RANTES protein mediate inflammation and cellular degradation in the cerebellum during P. yoelii 17XL malaria.

Activation requirements for CD4+ T cells differing in CD45R expression

Article

Nov 1992

Murine CD4+ T cells can be subdivided into naive and memory T cells based on surface phenotype, on recall response to Ag, and on differences in activation requirements. Furthermore, several studies have shown that two signals are required for CD4+ T cell activation; one signal is provided by occupancy of the TCR and the other signal is provided by the APC. In this report, analysis of naive and memory CD4 T cells, separated on the basis of CD45 isoform expression, has shown that their requirements for two signals differ. Activation of memory CD4 T cells to proliferate and secrete IL-2/IL-4 only required occupancy of the TCR complex, whereas activation of naive CD4 T cells required an APC-derived signal as well. Moreover, the signal induced by anti-CD3 antibodies differs from the signal provided by anti-V beta cross-linking of the TCR because both antibodies activate memory CD4 T cells but only anti-CD3 activates naive CD4 T cells. Together these data suggest that the consequence of stimulation through the TCR/CD3 signal complex differs between memory and naive CD4 T cells.

Functional interactions of T cells with endothelial cells: the role of CD40L-CD40-mediated signals

Article

Full-text available

Jan 1996

Chapter 1. Lymphocyte Adhesion Molecules: Role in Cell Adhesion and Intercellular Communication

Chapter

Dec 1994

Mitogenic and Non-Mitogenic Induction of Lymphocytic Invasion: Dual Parameter flow Cytometric Analysis

Chapter

Jan 1996

It has been well established that mitogenic activation produces profound alterations in the migratory behavior of lymphocytes. Activated, blastic T and B cells down-regulate homing receptors which mediate adhesion to the high endothelium of peripheral lymph nodes (1–3). Concurrently, they up-regulate adhesion receptors for extracellular matrix components and for ligands typical of inflammatory endothelium (4–6). The result is a change in traffic patterns, from extravasation into lymph nodes to extravasation into nonlymphoid tissue, especially inflamed tissue (7,8). After the immune response terminates, memory cells, the long-lived residual progeny of the blasts, remain. Memory helper T cells, even when not proliferating, retain the non-homing, inflammation-seeking behavior of the parental blasts (9–11). There are indications that other memory lymphocyte subsets follow similar trends (12,13).

Role of CD8+ αβ T Cells in Respiratory Infections Caused by Sendai Virus and Influenza Virus

Chapter

Jan 1993

Current understanding of the part played by CD8+αβ T cells in respiratory virus infections is based on findings from a spectrum of approaches. These include direct analysis of the patterns of lymphocyte involvement in the virus-infected lung (McDermott et al., 1987; Pena-Cruz et al., 1989; Allan et al., 1990; Openshaw, 1991; Eichelberger et al., 1991b), adoptive transfer experiments utilizing either bulk immune T-cell populations or cell lines (Leung and Ada, 1992; Lukacher et al., 1984, 1986; Ada and Jones, 1986; Taylor and Askonas, 1986; Kast et al., 1986; Cannon et al., 1987; Askonas et al., 1988; Mackenzie et al., 1989), in vivo depletion with monoclonal antibody (mAb) (Lightman et al., 1987; Waldmann, 1989; Allan et al., 1990; Eichelberger et al., 1991b), and the use of H-2 mutant or genetically manipulated mice (de Waal et al., 1983; Eichelberger et al., 1991b).

Adhesion Molecules involved in Leukocyte-Endothelial Cell Interactions

Chapter

Jan 1994

It has long been recognized that the successful extravasation of leukocytes through blood vessel walls into perturbed tissues is central to the progression of inflammatory reactions. The last decade has seen a growing awareness of the basic mechanisms underlying leukocyte emigration and, in particular, the importance of the adhesion molecules which facilitate interactions between leukocytes and vascular endothelial cells (EC). The considerable efforts expended in understanding cell adhesion have yielded not only a rich harvest of new molecules but have also revealed a complex web of interactions between them. The molecules involved in inflammation are reviewed in detail elsewhere 1, and this chapter will concentrate upon those molecules of particular relevance to interactions between leukocytes and EC.

Human endothelial cell presentation of antigen and the homing of memory/effector T cells to skin

Conference Paper

Jan 2001

Dermal microvascular endothelial cells (ECs) form a continuous lining that normally bars blood-borne T lymphocytes from entering the skin, but as part of the response to foreign antigen, dermal ECs undergo alterations. in their surface proteins so as to provide signals to circulating T cells that lead to their activation and recruitment. Several observations suggest that human dermal microvascular ECs may help initiate cutaneous Immune reactions by presentation of cognate antigens to circulating T memory cells: (1) antigen-specific inflammatory responses in the skin, as in other organs, involve accumulation of memory and effector T cell populations that are enriched in cells specific for the eliciting antigen; (2) recall responses to intradermal protein antigens in the skin start very rapidly within two hours of challenge; (3) dermal microvascular ECs in humans and other large mammals basally display high levels of class I and class II MHC molecules, the only known purpose of which is to present antigenic peptides to lymphocytes; (4) the lumen of dermal capillaries are narrower than the diameter of circulating T cells, ensuring surface contact; and (5) cultured human ECs effectively present antigens to resting memory T cells isolated from the circulation. Upon contact with activated T cells or their secreted products (cytokines), dermal ECs themselves become activated, increasing their capacity to recruit memory and effector T cell populations in an antigen-independent manner. Specifically, activated ECs express inducible leukocyte adhesion molecules such as E-selectin, ICAM-1, and VCAM-1; and several lines of evidence, including neutralizing antibody experiments and gene knockouts, have supported a role of these molecules in T cell recruitment. Dermal EC's have unique expression patterns of adhesion molecules that can determine the subsets of memory T cells that are recruited into the skin. For example, slow internalization of E-selectin allows more persistent expression of this protein on the surface of dermal ECs, favoring interactions with CLA-1(+) T cells.. VCAM-1 expression, normally confined to venular EC may extend to capillaries within the dermal papillae and contribute to epidermal inflammation, recruiting alpha (4)beta (7) integrin-expressing T cells that also express the cadherin-binding integrin alpha (E)beta (7). New models involving transplantation of normal and genetically modified human dermal ECs into immunodeficient mice may be used to further explore these properties.

Immunobiology, Diagnosis, and Treatment of Pancreas Graft Rejection

Chapter

Jan 2004

Rainer W. G. Gruessner

The immunologic basis of graft rejection was established by Medawar1,2 in the 1940s and 1950s. Since then, remarkable progress has been made in understanding the complexity of the rejection process and developing strategies to abrogate its occurrence. Over the last 20 years, it has become apparent that the immunobiology of pancreas allograft rejection is not different, by and large, from that of other types of solidorgan transplants. Some evidence, however, suggests that the mechanisms of rejection might be slightly different for exocrine vs endocrine pancreatic tissue.

Mast cell in immune function and leukocyte recruitment

Article

Apr 2013

A.F. De Felice

This chapter reviews published evidence that the mast cell (MC), beyond being among the first responders in host defense and the primary effector cell in Type I hypersensitivity responses to environmental allergens, also participates in more protracted, even chronic, inflammations, and can influence the natural history of such. As a legitimate immune effector cell, MCs can exacerbate a variety of hypersensitivity responses, many of which are auto-immunopathies, by regulating other immune and antigen-presenting cells through humoral and direct cell-cell contact, and by facilitating recruitment of leukocyte in early and, especially, late-phases of immune and other inflammatory reactions. Mast cells recruit leukocytes by promoting expression of adhesion molecules that effect critical leukocyte-endothelial cell interactions enabling leukocytic infiltration. These prominently include leukocyte and intercellular adhesion molecules (E-selectin; ICAM-1) induced by TNF-a, and interleukin-1 released from activated MC and other cells. Evidence that some cytokines or other MC or leukocyte mediators have prominent anti-inflammatory activity which limits immune injury and promotes tissue repair and re-modeling is also cited. Histamine, proteases, TNF-a and leukotrienes are selected as some MC mediators that have been monitored in autoimmune and other inflammatory conditions. The prospect remains speculative that these or other products of activated MC, even proteases unique to such cells, could be useful markers of clinical disease ancillary to diagnosis and follow-up.

Chronic Rejection in the Aorta Transplantation Model

Article

Jan 1999

Rob A. Geerling

Since the first successful heart transplantation in man in 1967, cardiac transplantation has become an accepted form of therapy for certain types of end-stage heart diseases.! After transplantation of a cadaveric heart graft, the immune system of the recipient will come in contact with donor cells. As clinical heart transplantation is an allogeneic type of grafting (genetically non-identical members of the same species), an immune response, which may result in graft rejection, will be inevitable. Of the foreign antigens that may be recognized by the immune system of the recipient, the major histocompatibility complex (MHC), in man the human leucocyte antigen (HLA) system, is the most important.2 - 4 In short, the HLA system consists of a group of closely linked genes, located on the short arm of chromosome-6 and divided in three regions, which encode class I and class II cell surface glycoproteins and several components of the complement system respectively. Class I is expressed on the surfaces of all nucleated cells. Class II is mainly expressed on antigen presenting cells (e.g, macrophages, dendritic cells, and B cells). After transplantation, T cell receptors of recipient CD4+ cells are able to recognize HLA class II, resulting in activation of type I and type 2 helper (THI and TH2) cells.' The activated THI cell start to preferentially synthesize and release interleukin-2 (IL-2) and interferon-gamma (IFN-y). IL-2 stimulates CD8+ T cells to develop into mature cytotoxic effector cells. Binding of these cells to donor HLA class I may result in graft cell lysis. IFN-y is responsible for the activation of macrophages, which are believed to be cytotoxic to graft cells. TH2 cells preferentially secrete IL-4, IL-6 and IL-IO. These interleukins are growth and differentiation factors for B cells. Activation of B cells may result in maturation into plasma cells with a11ospecific antibody production. Clinically, the two most important types of rejection that may occur after transplantation are acute and chronic rejection.

Respuesta inmunitaria celular cutánea. Un esquema fisiológico, patológico y farmacológico

Article

Mar 2005

Allograft rejection: The role played by adhesion molecules

Article

Jul 1994
Transplant Rev

Adhesion molecules and central nervous system inflammation

Article

Jun 1992
Semin Neurosci

From the ever-changing acronyms and the increased grouping of molecules with different acronyms into single units, it is apparent that a general consensus on the classification of adhesion molecules has yet to be achieved. This state of flux notwithstanding, three major groups (superfamilies) of adhesion molecules exist (the immunoglobulin, integrin and selectin superfamilies), with several less well-defined groups (e.g. addressins and cadherins) waiting in the wings, not yet granted superfamily status. Just as many of the immune system-related molecules play crucial roles in leukocyte traffic within lymphoid organs, it is now apparent that the same molecules are involved in immune system cell traffic and immune-mediated damage within non-lymphoid tissues like the central nervous system (CNS). For example, inflammation is the key in lesion development in some CNS diseases and data are presented here which strongly implicate adhesion molecules in immune cell trafficking to the CNS. Although a CNS-specific homing molecule or ligand has not yet been found, some of the vascular addressins constitutive for lymphoid tissues seem to display some site-specificity during the CNS autoimmune condition experimental allergic encephalomyelitis. Understanding the molecular mechanisms underlying CNS inflammation might lead to new therapeutic strategies, e.g. treatment with anti-adhesion molecule antibodies and antibodies against soluble mediators, such as cytokines, that modulate adhesion-molecule expression. Such strategies should have broad-ranging applicability.

Molecular mimicry: Basis for autoimmunity

Article

Aug 2000

Sheshadri Narayanan

Structural similarity between a viral protein and a self-component can trigger an autoimmune response, which is the basis of molecular mimicry. Alternatively an invading virus can induce an inflammatory response which in turn can initiate an attack by hitherto dormant T cells on a specific self-antigen, a phenomenon which is referred to as Bystander Activation. Several viruses share amino acid sequences with target self-proteins. A widely studied viral interaction is the structural mimicry of a small portion of coxsackie virus to a specific region of the enzyme glutamic acid decarboxylase (GAD) which is expressed by the β cells of the islet of Langerhans in the pancreas leading to the destruction of insulin producing cells and the onset of Type I insulin dependent diabetes mellitus (IDDM). Knowledge of specific epitopes in GAD susceptible to autoimmune attack can permit devising therapeutic strategies for the prevention and suppression of IDDM.

Anti-adhesive therapeutics: A new class of anti-inflammatory agents

Article

Mar 2008

Intercellular adhesion plays a discrete part in a co-ordinated sequence of events which lead to the pathology of inflammatory disease. The last decade has seen an explosion of information aimed at providing more detail on the mechanism of cell adhesion which essentially involves a number of signal transductions resulting in the migration of white blood cells to the site of infection. Over-recruitment of these cells can cause injury to the normal tissues as observed in the case of septic shock-reperfusion injury. Cell-cell interaction also takes place in metastasis - the migration and adherence of malignant tumour cells. Inhibition or control of these pathophysiological disorders is possible by introducing therapeutics that would block or modulate the adhesion process.

Restricted heterogeneity of T cell receptor variable alpha chain transcripts in hearts of Chagas’disease cardiomyopathy patients

Article

Oct 2007
PARASITE IMMUNOL

The role of autoimmunity in the pathogenesis and progression of heart lesions in the chronic phase of Chagas’disease is controversial. In the absence of parasites in situ, the T cell infiltrate seen in heart lesions may be the primary determinant of tissue damage ultimately leading to heart failure and death. We used the polymerase chain reaction to amplify each known T cell receptor (TCR) Vα and Vβ subfamily-specific sequence in transcripts derived from heart samples obtained from Chagas’cardiomyopathy patients. The average number of TCR Vα. subfamilies (7·1 per tissue sample) was significantly lower than that for TCR Vβ subfamilies (15·1 per sample). The average percentage of tissue samples positive per TCR Vα. and Vβ subfamily was respectively 39·6% vs. 73·5%. These data suggest that, in Chagas’heart lesions, the detectable TCR Vα. repertoire is significantly narrower than TCR Vβ repertoire. On the other hand, in normal heart tissue, diversity of Vα. and Vβ TCR is similar among the scarce circulating T cell population. Such evidence of restricted TCR V region repertoire has been described in experimental and human autoimmune diseases. Our results are consistent with the possibility that T cells responsible for heart damage in chronic Chagas’cardiomyopathy may be recognizing a few heart-specific antigenic targets.

Human Endothelial Cell Presentation of Antigen and the Homing of Memory/Effector T Cells to Skin

Article

Sep 2001
ANN NY ACAD SCI

Dermal microvascular endothelial cells (ECs) form a continuous lining that normally bars blood-borne T lymphocytes from entering the skin, but as part of the response to foreign antigen, dermal ECs undergo alterations in their surface proteins so as to provide signals to circulating T cells that lead to their activation and recruitment. Several observations suggest that human dermal microvascular ECs may help initiate cutaneous immune reactions by presentation of cognate antigens to circulating T memory cells: (1) antigen-specific inflammatory responses in the skin, as in other organs, involve accumulation of memory and effector T cell populations that are enriched in cells specific for the eliciting antigen; (2) recall responses to intradermal protein antigens in the skin start very rapidly within two hours of challenge; (3) dermal microvascular ECs in humans and other large mammals basally display high levels of class I and class II MHC molecules, the only known purpose of which is to present antigenic peptides to lymphocytes; (4) the lumen of dermal capillaries are narrower than the diameter of circulating T cells, ensuring surface contact; and (5) cultured human ECs effectively present antigens to resting memory T cells isolated from the circulation. Upon contact with activated T cells or their secreted products (cytokines), dermal ECs themselves become activated, increasing their capacity to recruit memory and effector T cell populations in an antigen-independent manner. Specifically, activated ECs express inducible leukocyte adhesion molecules such as E-selectin, ICAM-1, and VCAM-1; and several lines of evidence, including neutralizing antibody experiments and gene knockouts, have supported a role of these molecules in T cell recruitment. Dermal ECs have unique expression patterns of adhesion molecules that can determine the subsets of memory T cells that are recruited into the skin. For example, slow internalization of E-selectin allows more persistent expression of this protein on the surface of dermal ECs, favoring interactions with CLA-1+ T cells. VCAM-1 expression, normally confined to venular EC may extend to capillaries within the dermal papillae and contribute to epidermal inflammation, recruiting 4β7 integrin-expressing T cells that also express the cadherin-binding integrin Eβ7. New models involving transplantation of normal and genetically modified human dermal ECs into immunodeficient mice may be used to further explore these properties.

Inflammation and glomerular injury

Article

Jul 1994

ExperimentalModels of Cerebral Malaria

Chapter

Full-text available

Malaria remains a major global health problem and cerebral malaria is one of themost serious complications of this disease. Recent years have seen important advances in our understanding of the pathogenesis of cerebralmalaria. Extensive analysis of tissues and blood taken frompatients with cerebralmalaria has been complimented by the use of animal models to identify specific components of pathogenic pathways. In particular, an important role for CD8+ T cells has been uncovered, as well divergent roles for members of the tumor necrosis factor (TNF) family of molecules, including TNF and lymphotoxin alpha. It has become apparent that theremay bemore than one pathogenic pathway leading to cerebral malaria. The last few years have also seen the testing of vaccines designed to target malaria molecules that stimulate inflammatory responses and thereby prevent the development of cerebral malaria. In this review, we will discuss the above advancements, as well as other important findings in research into the pathogenesis of cerebral malaria. As our understanding of pathogenic responses to Plasmodium parasites gathers momentum, the chance of a breakthrough in the development of treatments and vaccines to prevent death fromcerebralmalaria have become more realistic.

Liver endothelial cells: participation in host response to lymphoma metastasis

Article

Jan 1996

Interactions between metastasizing tumor cells and host cells in target organs determine the outcome of metastasis. This review discusses the dual role of activated host endothelial cells in the metastatic process. On one hand, the upregulation of the expression of particular adhesion molecules leads to increased tumor cell binding, and the stimulation of angiogenesis provides the vascular support for the growth of already established metastases. On the other hand, endothelial cells can contribute to host anti-metastatic responses, e.g. by production of the cytotoxic molecule nitric oxide (NO) from arginine with the help of the inducible nitric oxide synthase (iNOS). Using a well-characterized ESbL-lacZ mouse T lymphoma model with a typical three phasic growth profile, we showed during the period of growth retardation a stimulation of NO production by ex vivo isolated liver sinusoidal endothelial cells. The induction of NO synthesis in liver endothelial cells did not require the presence of Kupffer cells and appeared to be stimulated by and dependent on mature T lymphocytes. A breakdown of this NO synthesis coincided with the second tumor expansion phase.

Expression of adhesion molecules by endothelial cells of early human decidua

Article

Jul 1993

The expression of adhesion molecules by endothelial cells (EC) of early human decidua was studied with monoclonal antibodies and the immunoperoxidase technique. Although E-selectin, INCAM-110 and VCAM-1 were poorly detected on decidual EC, ICAM-1, P-selectin and DR antigens were highly expressed by these cells, some of which showed high endothelial venule-like morphology. Our results suggest that decidual EC are activated, and are probably involved in the active recruitment of leucocytes.

Catecholamine Modulation of Lymphocyte Homing to Lymphoid Tissues

Article

Dec 1997

Lymphocyte migration is an essential process for immune surveillance and for promoting cell-cell interactions necessary to generate an immune response. This report examined whether catecholamine prestimulation would alter the pattern of lymphocyte homing to spleen and lymph nodes in mice as determined by tracking fluorescently labeled cells. The results of cell sorter analysis showed that catecholamine-pretreated cells had increased accumulation in spleen and lymph nodes 1 and 2 h after i.v. injection. In addition, microscopic analysis showed that labeled cells migrated from the splenic red pulp to T-cell regions of the white pulp over a 2-h time course. Within the lymph nodes, labeled cells localized predominantly to the pericortex. Additional studies examined the migration of lymphocytes to lymphoid tissues of NGF-transgenic mice that have sympathetic hyperinnervation of spleen and peripheral lymph nodes. In contrast to the studies above, migration of T-cells from control mice to lymphoid tissues of the hyperinnervated mice was not different than that in control mice in most tissues. The accumulation of lymphocytes in lymphoid tissues is a balance between the influx of newly migrated cells and efflux back into the circulation. The studies in this report lend support to other studies showing catecholamine modulation of lymphocyte migration and homing, but it is a complex process about which much has yet to be understood.

The Molecular Immunology of Acute Rejection: An Overview

Article

Mar 1993
TRANSPL IMMUNOL

Transplantation immunology began as an empirical science but can now take its place as a discipline with a strong theoretical and molecular basis. The Laws of Transplantation, formulated at the beginning of this century by Little and Tyzzer, can be paraphrased as 'Isografts succeed; allografts fail'. As the century closes, the molecular basis of those laws is emerging. Achieving the full potential of the advances in immunology requires that transplant clinicians learn the molecular basis for their art and participate in the development and testing of new hypotheses; and that immunologists assess their ideas against the real world of the transplant recipient. This review will highlight areas of recent progress in transplant immunology and point out areas where clinical events remain poorly explained. Because many excellent reviews are available, we will primarily quote articles after 1990. Our approach to transplantation biology at the molecular level is summarized in Table 1.

Enterovirus Infection, CXC Chemokine Ligand 10 (CXCL10), and CXCR3 Circuit: A Mechanism of Accelerated -Cell Failure in Fulminant Type 1 Diabetes

Article

Full-text available

Aug 2009

Fulminant type 1 diabetes is characterized by the rapid onset of severe hyperglycemia and ketoacidosis, with subsequent poor prognosis of diabetes complications. Causative mechanisms for accelerated beta-cell failure are unclear. Subjects comprised three autopsied patients who died from diabetic ketoacidosis within 2-5 days after onset of fulminant type 1 diabetes. We examined islet cell status, including the presence of enterovirus and chemokine/cytokine/major histocompatibility complex (MHC) expressions in the pancreata using immunohistochemical analyses and RT-PCR. Immunohistochemical analysis revealed the presence of enterovirus-capsid protein in all three affected pancreata. Extensive infiltration of CXCR3 receptor-bearing T-cells and macrophages into islets was observed. Dendritic cells were stained in and around the islets. Specifically, interferon-gamma and CXC chemokine ligand 10 (CXCL10) were strongly coexpressed in all subtypes of islet cells, including beta-cells and alpha-cells. No CXCL10 was expressed in exocrine pancreas. Serum levels of CXCL10 were increased. Expression of MHC class II and hyperexpression of MHC class I was observed in some islet cells. These results strongly suggest the presence of a circuit for the destruction of beta-cells in fulminant type 1 diabetes. Enterovirus infection of the pancreas initiates coexpression of interferon-gamma and CXCL10 in beta-cells. CXCL10 secreted from beta-cells activates and attracts autoreactive T-cells and macrophages to the islets via CXCR3. These infiltrating autoreactive T-cells and macrophages release inflammatory cytokines including interferon-gamma in the islets, not only damaging beta-cells but also accelerating CXCL10 generation in residual beta-cells and thus further activating cell-mediated autoimmunity until all beta-cells have been destroyed.

The role of adhesion molecules in endothelial cell accessory function

Article

Dec 1992

Alloantigenicity of human endothelial cells. 1. Frequency and phenotype of human T helper lymphocytes that can react to allogeneic endothelial cells

Article

Jul 1992

To determine the relative ability of allogeneic endothelial cells to stimulate helper T lymphocytes (HTL), human PBMC or purified T cells were incubated in conventional lymphocyte microcultures or in limiting dilution microcultures with allogeneic human umbilical vein endothelia (HUVE), with cytokine-treated allogeneic HUVE, or with allogeneic peripheral blood monocytes. These cultures were tested for IL-2 production as an index of HTL stimulation. Dose-response studies in conventional lymphocyte cultures indicated that allogeneic monocytes were better than allogeneic HUVE at stimulating IL-2 production. Limiting dilution analyses revealed that untreated HUVE and TNF-treated HUVE stimulated small numbers of HTL (approximately 1 HTL/30,000 PBMC), whereas 5 to 10 times more HTL were stimulated by IFN-gamma-treated HUVE and 10 to 20 times more HTL were stimulated by allogeneic monocytes. Serologic deletion studies revealed that most of the high frequency HTL responding to IFN-gamma-treated HUVE were CD4+, whereas most of the low frequency HTL responding to nontreated HUVE or to TNF-treated HUVE were CD8+. Interestingly, mAb to MHC class I and class II molecules, which significantly impaired HUVE-induced proliferation, caused little interference with HUVE-induced IL-2 production. Finally, polymerase chain reaction analysis demonstrated that untreated allogeneic HUVE cells could stimulate PBMC to produce mRNA for IFN-gamma, as well as for IL-2. These data demonstrate the following hierarchy of allogeneic stimulatory capacity for human HTL: monocytes greater than IFN-gamma-treated HUVE much greater than TNF-treated HUVE = nontreated HUVE. Further, these data suggest that non-activated allogeneic endothelial cells can initiate immune responses by inducing IL-2 and IFN-gamma. Because IFN-gamma can induce MHC class II expression by the endothelial cells, this could recruit large numbers of CD4+ T cells for IL-2 production.

Differential expression of VLA-α4 and VLA-β1 discriminates multiple subsets of CD4+CD45R0+ "memory" T cells

Article

Jan 1993

Given the importance of adhesion in T cell development, we have undertaken systematic flow cytometric analysis of CD4 T cells to determine relationships between the developmentally regulated marker CD45R0 and adhesion receptors (five VLA integrin chains). The most important findings are that: 1) expression of alpha 3, alpha 5, and alpha 6 are closely coregulated with beta 1 on CD4 cells, while regulation of VLA-alpha 4 is quite discordant. 2) CD45R0- cells, generally understood to be naive cells, have low homogeneous expression of VLA-alpha 3, VLA-alpha 4, VLA-alpha 5, VLA-alpha 6, and beta 1 integrin chains; studies of cord blood CD4 cells confirm the low homogeneous expression of alpha 4 and beta 1 on naive cells. 3) In marked contrast, CD45R0+ cells, generally understood to be memory cells, show not only an overall increase in expression of these integrins (relative to CD45R0- cells) but also heterogeneity. Dramatic heterogeneity is revealed when the markers VLA-alpha 4 and beta 1 are analyzed together. Many CD45R0+ cells show increased levels of both VLA-alpha 4 and VLA-beta 1; however, some have increased levels principally of either VLA-beta 1 or VLA-alpha 4. We hypothesize that T cells becoming memory cells in different microenvironments specialize their integrin phenotype, thereby acquiring distinctive functional and homing capacities; in this process, VLA-4 (CD49d) appears to play a unique role.

Random entry of circulating lymphocyte subsets into peripheral lymph nodes and Peyer's patches: No evidencein vivo of a tissue-specific migration of B and T lymphocytes at the level of high endothelial venules

Article

Sep 1992

Lymphocytes continuously migrate through the body and thus immune competent cells are constantly delivered to most tissues. They interact with high endothelial venules (HEV) via specific homing receptors and vascular addressins, and these molecules seem to be the reason for a preferential homing of B lymphocytes into Peyer's patches and of T lymphocytes into peripheral lymph nodes. When lymphocytes derived from lymph node cell suspensions were applied in the in vitro lymphocyte/endothelium binding assay, the well‐known preference of mouse lymph node B lymphocytes for Peyer's patch HEV compared to peripheral lymph node HEV was confirmed in the rat (2.8 times). When in the same in vitro assay thoracic duct lymphocytes (TDL) were used this preference was far less obvious (1.4 times). However, by injecting rat TDL intravenously and by tracing them directly in HEV, B, T, CD4 ⁺ and CD8 ⁺ lymphocytes are seen to enter Peyer's patches and peripheral lymph nodes in vivo without preference. Thus, in contrast to lymphocytes from lymph node cell suspensions, no evidence was found of a tissue‐specific migration of thoracic duct B, T, CD4 ⁺ and CD8 ⁺ lymphocytes at the HEV level. This finding demonstrates the importance of considering both experimental conditions and the cell source used when investigating lymphocyte traffic.

Migration of Activated Lymphocytes

Article

Feb 1993
CURR TOP MICROBIOL

When lymphocytes leave the primary lymphoid organs as mature but naive cells, they enter the recirculating pool, moving continuously from the blood into the secondary lymphoid organs, spleen, lymph nodes and Peyer’s patches, from which they return, directly or via efferent lymph, back to the blood (Gowans and Knight 1964). In addition, some of the cells also pass through nonlymphoid tissues, such as lung, liver, gut wall or skin, from which they return into circulation either directly or via afferent lymph/lymph nodes/efferent lymph (Fig. 1).

Endothelial activation and circulating vascular adhesion molecules in alcoholic liver disease

Article

Mar 1994

Alcoholic hepatitis is characterized by hepatocyte necrosis associated with infiltration of the liver parenchyma by neutrophils. The mechanisms responsible for recruiting neutrophils to the liver are unknown. We report high circulating levels and tissue expression of the endothelial adhesion molecule E-selectin in alcoholic hepatitis. Because expression of E-selectin is involved in neutrophil transmigration into inflamed tissue, it may play a crucial role in the recruitment of neutrophils to the liver in alcoholic hepatitis. By contrast, we detected high levels of vascular cell adhesion molecule-1, the endothelial counter-receptor for the lymphocyte adhesion molecule very late antigen-4, in alcoholic cirrhosis, which is associated with a predominantly mononuclear cell infiltrate. Both diseases were associated with high levels of circulating intercellular adhesion molecule-1, which is released by activated lymphocytes, providing further evidence of immune activation in alcoholic liver disease.

Substance P and adrenocorticotropic hormone do not affect T-lymphocyte adhesion to vascular endothelium or surface expression of adhesion receptors

Article

Mar 1994
Int J Immunopharm

Substance P (SP) and adrenocorticotropic hormone (ACTH) are peptides that have been shown to have both neurological and immunological effects. Because of the demonstrated effects upon immune function, we examined the effects of these peptides on T-lymphocyte adhesion to vascular endothelium and surface adhesion receptor expression. Neither the adhesion assays nor the expression assays showed any statistically significant effect of SP (10 microM) or ACTH (1 microM) for any incubation period used. We conclude that, while SP and ACTH have a variety of immunomodulatory effects, direct modulation of T-lymphocyte adhesion to vascular endothelium is probably not one of them.

Characterization and functional analysis of the expression of vascular adhesion molecules in human papillomavirus-related disease of the cervix

Article

Sep 1994

Cervical intraepithelial neoplasia (CIN) is associated with changes in local immune cell populations, although the role of vascular adhesion molecules in mediating such changes by controlling the traffic of mononuclear cells to the cervix has not been investigated previously. The authors used immunohistochemistry to examine the expression of three vascular adhesion molecules--ICAM-1, VCAM-1 and E-selectin--in the normal cervix and in biopsies of CIN Grade 1 (CIN-1) (low grade squamous intraepithelial lesions [LG-SIL]) and CIN-2/3 (high grade squamous intraepithelial lesions[HG-SIL]). In addition, the authors examined the functional role of these molecules by adapting the frozen section adhesion assay of Stamper and Woodruff to investigate in vitro the molecular basis of the interaction between cervical endothelial cells and activated T-lymphocytes. Whereas there was no difference in adhesion molecule expression between normal cervix and CIN-1 (LG-SIL), all three molecules investigated were significantly up-regulated in CIN-2/3 (HG-SIL), an observation that correlated with an enhanced ability of stromal endothelial cells in CIN-2/3 (HG-SIL) biopsies to bind activated peripheral blood lymphocytes in vitro. Monoclonal antibodies blocking ICAM-1 function were able to reduce such adhesion significantly in three of three experiments, and antibodies blocking VCAM-1 produced a significant reduction in one of three experiments. No inhibition was seen with antibodies against E-selectin. The enhanced expression of vascular adhesion molecules in CIN-2/3 (HG-SIL) appears to be functionally important in enabling the local recruitment of immunocompetent cells and supports the notion of a local antineoplastic immune response in high grade cervical intraepithelial lesions.

Suppressive effect of basic fibroblast growth factor on transendothelial emigration of CD4(+) T-lymphocyte

Article

Oct 1994

The effect of basic fibroblast growth factor (b-FGF), one of the commonest angiogenic factors in various cancer types, on lymphocyte adhesion and transmigration across the endothelial cell monolayer was investigated using human umbilical vein-derived endothelial cells (HUVEC) and type I collagen gel. Forty-eight h exposure of HUVEC with 2 ng/ml b-FGF significantly decreased the basal adhesion of lymphocytes to endothelial cells. The decrease ratio is further enhanced by the addition of shear stress in this assay system. When HUVEC was stimulated for the last 24 h with optimal conditions of recombinant interleukin 1 beta, the percentages of transmigration as well as adhesion were also decreased significantly by the presence of b-FGF. The expression of intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 was down-regulated by b-FGF exposure in both resting and activated conditions by recombinant interleukin 1 beta, supposedly the main reason for this phenomenon. The migrating cells across b-FGF-stimulated HUVEC contained a markedly lower percentage of CD4(+) T-cells than those across non-treated HUVEC, although the 4B4(+)/2H4(+) ratio in CD4(+) T-cell populations did not differ significantly. These facts suggest that the presence of b-FGF in the angiogenic area suppresses lymphocyte emigration, especially that of CD4(+) T-cells, and thus causes insufficient helper function in local immune response. This effect of b-FGF was possibly one of the critical mechanisms by which cancer cells escape from the host immune reactions in the angiogenic stage of tumor development.

The proliferative response of human T cells to allogeneic IFN-γ-treated endothelial cells is mediated via both CD2/LFA-3 and LFA-1/ICAM-1 and -2 adhesion pathways

Article

Feb 1993
TRANSPL IMMUNOL

We studied the proliferative response of purified human peripheral blood T lymphocytes (contaminated with less than 0.1% monocytes) to allogeneic MHC class II molecules expressed by endothelial cells (EC) or fibroblasts (FB). In vitro expression of MHC class II molecules was induced by gamma-interferon (IFN-gamma) treatment. The MHC class II expression levels after IFN-gamma treatment on both cell types were comparable. No T cell proliferation was found in the presence of either untreated or IFN-gamma-treated FB, and a marginal proliferation in the presence of untreated EC. IFN-gamma-treated EC, however, were able to induce significant T cell growth. The previously established role of MHC class II molecules in allogeneic T cell proliferation was confirmed in inhibition experiments with monoclonal antibody (mAb) against MHC class II or CD4. In this model, we tested the involvement of a number of adhesion molecules by adding mAbs to cocultures of T cells and IFN-gamma-treated EC. Monoclonal antibodies directed against CD31, CD26, B7/BB1, E-selectin, CD44, VLA-4 alpha-chain and VCAM-1 had no effect, whereas moderate inhibition was observed with anti-VLA-beta-chain and anti-LFA-3. A distinct inhibition of T cell proliferation was observed with mAbs directed against LFA-1, CD2, or a combination of anti-ICAM-1 and -2. Combinations of mAbs directed against T cell adhesion molecules (LFA-1, CD2, VLA-4) or EC adhesion molecules (ICAM-1, and -2, LFA-3, VCAM-1) were able to block T cell proliferation for 100 and 80% respectively. We conclude that CD2/LFA-3 and LFA-1/ICAM interactions are crucially involved in allogeneic T cell/EC interactions.

Synthesis and release of platelet-activating factor by human vascular endothelial cells treated with tumor necrosis factor or interleukin 1 alpha.

Article

Full-text available

Aug 1988

Human endothelial cells synthesize large amounts of platelet-activating factor (PAF) after 30-min treatment with recombinant tumor necrosis factor (TNF). Synthesis of PAF peaks at 4-6 h, whereas in endothelial cells treated with interleukin 1 alpha (IL-1) it peaks at 8-12 h. More than twice as much PAF is synthesized in response to optimal concentrations of TNF than in response to IL-1. However, PAF synthesis is stimulated by lower molar concentrations of IL-1 than TNF. About 30% of PAF produced in response to either TNF or IL-1 is released into the medium, whereas approximately 70% remains cell-associated. Experiments with labeled precursors show that PAF is synthesized de novo in response to TNF. This activity of TNF is inhibited by treating endothelial cells with the inhibitors of protein or RNA synthesis cycloheximide or actinomycin D. This finding may be explained by the observation that TNF induces in endothelial cells an acetyltransferase required for PAF synthesis. The induction of this enzymatic activity precedes the peak of PAF synthesis in TNF-treated cells. After prolonged incubation with either TNF or IL-1, endothelial cells no longer respond to the same monokine, but are still capable of producing PAF when treated with the other monokine. The finding that these monokines do not show reciprocal tachyphylaxis in endothelial cells may be explained by their binding to different receptors. In cells treated simultaneously with different concentrations of TNF and IL-1, PAF synthesis is stimulated in an additive rather than synergistic way. This suggests that PAF is synthesized by the same pathway in response to TNF or IL-1.

Tumor necrosis factor/cachectin interacts with endothelial cell receptors to induce release of interleukin 1

Article

Full-text available

Jun 1986

Peter P Nawroth

Tumor necrosis factor/cachectin (TNF) has been implicated as a mediator of the host response in sepsis and neoplasia. Recent work has shown that TNF can modulate endothelial cell hemostatic properties, suggesting that endothelium is a target tissue for TNF. This led us to examine whether endothelial cells have specific binding sites for TNF and augment the biological response to TNF by elaborating the inflammatory mediator, IL-1. Incubation of 125I-recombinant human TNF with confluent, cultured human umbilical vein endothelial cells resulted in time-dependent, reversible, and saturable binding. Binding was half-maximal at a TNF concentration of 105 +/- 40 pM, and at saturation 1,500 molecules were bound per cell. Heat-treated TNF, which is biologically inactive, did not bind to endothelium. In addition to surface binding, TNF induced the elaboration of IL-1 activity by endothelial cells in a time-dependent manner. Generation of IL-1 activity required protein synthesis and was half-maximal at a TNF concentration of 50 +/- 20 pM. IL-1 activity from TNF-treated endothelium could be adsorbed by an immobilized antibody to IL-1. Heat-treated TNF was ineffective in eliciting endothelial cell IL-1. These data indicate that TNF can bind specifically to endothelium and initiate a cascade of inflammatory and coagulant events on the vessel surface potentially central to the host response to neoplasia and sepsis.

Phenotypic and Functional Characterization of Lymphocytes That Bind Human Microvascular Endothelial Cells In Vitro Evidence for Preferential Binding of Natural Killer Cells

Article

Full-text available

Jun 1987

The microvascular endothelium has been postulated to be a critical target in the rejection of vascularized allografts. This study was undertaken to examine the ability of human sheep erythrocyte rosette forming lymphocytes (E-RFC) to form stable conjugates with microvascular endothelial cells (EC), and to assess whether a receptor-ligand interaction mediates this event. Human foreskin microvascular EC monolayers were used as targets of chromium-51-labeled E-RFC in a quantitative adherence assay. Binding was saturable, displaceable by unlabeled E-RFC, augmented by recombinant interleukin 1 (rIL-1) and inhibited by anti-LFA1 antibody. The Leu-11+ lymphocyte subset, known to be enriched for natural killer (NK) cells, bound preferentially. Only the EC-adherent lymphocyte fraction contained NK effectors, which lysed EC and classical NK targets. Thus, NK cells adhere to microvascular EC via a specific receptor-ligand interaction. The possibility exists that such binding occurs in recipients of vascularized allografts, representing the initial stage of graft rejection.

Interleukin 1 induces cultured human endothelial cell production of granulocyte-macrophage colony-stimulating factor

Article

Full-text available

Jan 1987

Monokine-stimulated endothelial cells are known to produce both burst- and colony-stimulating activities, but neither the nature of the monokine nor the hematopoietic growth factor(s) produced is known. We show by mRNA analysis that an immortalized line of human endothelial cells constitutively produce granulocyte-macrophage colony-stimulating factor. Furthermore, interleukin 1 and tumor necrosis factor induce early passage human umbilical endothelial cells to produce the same growth factor.

Synthesis of Prostaglandin I2 (Prostacyclin) by Cultured Human and Bovine Endothelial Cells

Article

Full-text available

Oct 1977

Cultured endothelial cells derived from human umbilical veins or bovine aorta produce a potent inhibitor of platelet aggregation. The inhibitor is synthesized from sodium arachidonate or or prostaglandin endoperoxides by a microsomal enzyme system. Tranylcypromine, a specific antagonist of prostacyclin synthetase, suppresses production of the inhibitor by endothelial cells. The inhibitor, which is ether extractable, has been identified using a two-step thin-layer radiochromatographic procedure and a synthetic prostaglandin I2 standard. With this procedure, we have shown that human and bovine endothelial cells convert sodium [3H]arachidonate to radiolabeled prostaglandin I2 and 6-keto-prostaglandin F1alpha, as wellas prostaglandin E2. Thus, endothelial cells may be non-thrombogenic in vivo because they synthesize and release prostaglandin I2, a potent inhibitor of platelet aggregation.

Lymphocyte homing into lymph nodes. In vitro demonstration of the selective affinity of recirculating lymphocytes for high-endothelial venules

Article

Full-text available

Oct 1976

An in vitro model is described for studying the interaction between lymphocytes and high-endothelial venules (HEV) of lymph nodes. Rat or mouse lymphocytes which were layered over fixed sections of syngeneic lymph nodes adhered selectively to the endothelium of HEV but did not bind to other vascular endothelia. Evidence is presented that adherence to HEV in vitro is a property of recirculating lymphocytes and not a characteristic of cells which are unable to home into lymph nodes in vivo.

Identification of surface proteins mediating adherence of CDH/CDI8 deficient lymphoblastotd cells to cultured human endolhelium

Article

Full-text available

Jul 1990

Patients with the severe form of leukocyte adhesion deficiency syndrome do not express the CD11/CD18 adhesion complex on any of their leukocytes. Nevertheless, their lymphocytes, unlike their phagocytes, emigrate to extravascular sites of inflammation, demonstrating that surface proteins other than CD11/CD18 can mediate lymphocyte adherence to endothelium. Using a B-lymphoblastoid cell line (B-LCL) established from a CD11/CD18-deficient patient and cultured human umbilical vein endothelial cells (HEC), we investigated the CD11/CD18-independent mechanism(s) of lymphocyte adherence to endothelium. Monoclonal antibodies directed to the alpha 4 polypeptide (CD49d) and the beta 1 polypeptide (CD29) of the lymphocyte VLA-4 integrin receptor (CD49d/CD29), and to vascular cell adhesion molecule-1 (VCAM-1) on the endothelial cell significantly inhibited the adherence of the CD11/CD18-deficient B-LCL to untreated HEC and to HEC treated with recombinant human tumor necrosis factor-alpha. We suggest that the interaction of the lymphocyte receptor VLA-4 with the endothelial ligand VCAM-1 induced by cytokines at sites of inflammation or immune reaction represents a CD11/CD18-independent pathway of lymphocyte emigration.

Picker, L. J., Michie, S. A., Rott, L. S. & Butcher, E. C. A unique phenotype of skin-associated lymphocytes in humans. Am. J. Pathol. 136, 1053

Article

Full-text available

Jun 1990

It has been proposed that the skin is a functionally unique compartment of the immune system, although little direct evidence supporting this hypothesis has been presented. Here we show that lymphocyte populations at cutaneous sites can be differentiated from otherwise similar populations at noncutaneous sites by their preferential expression of an epitope defined by the MAb HECA-452. This MAb recognizes a predominantly 200-kd cell-surface glycoprotein present on about 16% of peripheral blood T cells, including both CD4+ and CD8+ T cells (17% and 11% HECA-452+, respectively), as well as TCR-delta-bearing T cells (32%+). Most thymocytes (99%) lacked HECA-452 antigen expression, and essentially all the HECA-452+ peripheral blood T cells were found in the adhesion molecule high, CD45R low putative memory cell subset, findings suggesting that HECA-452 expression develops peripherally as a consequence of antigenic stimulation. However, the HECA-452 antigen is not a conventional activation antigen because it was not upregulated with mitogen stimulation of peripheral blood T cells. Most significantly, among 54 diverse specimens of normal/reactive lymphoid tissues and sites of chronic inflammation, there was a clear association of lymphocyte HECA-452 expression and cutaneous location. In extracutaneous sites (n = 38) only about 5% of lymphocytes within the T-cell areas of these tissues expressed this antigen, whereas in inflammatory skin lesions (n = 16), 85% were HECA-452+. The association of HECA-452 expression and cutaneous location was also seen in a series of T-cell lymphomas. The malignant cells of 16 of 18 cases of epidermotropic (patch/plaque) stage mycosis fungoides were HECA-452+, as well as 2 of 7 nonmycosis fungoides peripheral T-cell lymphomas in skin. In contrast, this antigen was not expressed in thymic (lymphoblastic) lymphomas (n = 14), nonepidermotropic (tumor) stage mycosis fungoides (n = 5), and noncutaneous peripheral T-cell lymphomas (n = 15). Among lymphocytes, the preferential expression of the HECA-452 determinant by cutaneous T cells supports the hypothesis that the skin constitutes a immunologically unique lymphoid tissue and suggests that this molecule may play a role in either lymphocyte homing to skin or in lymphocyte interactions with the epidermis.

Interleukin-1 is an autocrine regulator of human endothelial

Article

Full-text available

Oct 1990

Proliferation of endothelial cells is regulated through the autocrine production of growth factors and the expression of cognate surface receptors. In this study, we demonstrate that interleukin 1 (IL-1) is an inhibitor of endothelial growth in vitro and in vivo. IL-1 arrested growing, cultured endothelial cells in G1 phase; inhibition of proliferation was dose dependent and occurred in parallel with occupancy of endothelial surface IL-1 receptors. In an angiogenesis model, IL-1 could inhibit fibroblast growth factor-induced vessel formation. The autocrine nature of the IL-1 effect on endothelial proliferation was demonstrated by the observation that occupancy of cell-surface receptors by endogenous IL-1 depressed cell growth. The potential significance of this finding was emphasized by the detection of IL-1 in the native endothelium of human umbilical veins. A mechanism by which IL-1 may exert its inhibitory effect on endothelial cell growth was suggested by studies showing that IL-1 decreased the expression of high-affinity fibroblast growth factor binding sites on endothelium. These results point to a potentially important role of IL-1 in regulating blood vessel growth and suggest that autocrine production of inhibitory factors may be a mechanism controlling proliferation of normal cells.

A monoclonal anti-HEBFPP antibody with specificity for lymphocyte surface molecules mediating adhesion to Peyer's patch high endothelium of the rat.

Article

Apr 1986

Cell surface molecules involved in lymphocyte adhesion to high endothelial cell venules (HEV) of Peyer's patches (PP) have been studied in the rat by using a mouse monoclonal anti-HEBFPP (1B.2) antibody. We previously showed that rat thoracic duct lymph contains a high endothelial cell binding factor termed HEBFPP, which in vitro blocks lymphocyte binding sites of HEVPP but not HEVLN. Monoclonal 1B.2 antibody was produced by fusing P3U1 myeloma cells with spleen cells of a mouse immunized with this material. Immunoprecipitation studies with 125I surface-labeled rat thoracic duct lymphocytes (TDL) showed that the antibody recognized an 80-kilodalton protein. This antigen was present in the majority of TDL, spleen, LN, and PP cells but was found on few (5 to 10%) thymus and bone marrow cells (indirect immunofluorescence). Treatment of TDL with 1B.2 antibody blocked their ability to bind in vitro to HEVPP; antibody treatment did not interfere with TDL adhesion to HEVLN. Analysis of 1B.2 antigen isolated from lymph and detergent lysates of TDL by antibody-affinity chromatography showed that this material had the capacity to block lymphocyte binding sites of HEVPP but not HEVLN. In contrast, material with such blocking activity was not isolated from detergent lysates of thymocyte, a population deficient in HEV-binding cells. The results indicate that the 1B.2 antigen is a component of the lymphocyte surface recognition structure mediating adhesion to HEVPP and provide further evidence that distinct adhesion molecules of rat TDL mediate interaction with high endothelium of LN and PP.

Regulation of class II MHC molecules on human endothelial cells. Effects of IFN and dexamethasone.

Article

Jun 1988

Human vascular endothelial cells normally do not express class II MHC molecules in culture. IFN-gamma has been shown to induce expression of class I and class II MHC molecules on endothelial cell cultures from umbilical cord. We could detect these Ag by FACS analysis when endothelial cells were cultured for 3 days in the presence of 200 to 1000 U/ml of rIFN-gamma. Among the class II MHC molecules, HLA-DR and -DP but not -DQ were consistently induced. Addition of rIFN-alpha-D/A to IFN-gamma-treated cells inhibited the expression of class II MHC but not class I MHC molecules. Furthermore, the inhibition was more pronounced when IFN-alpha-D/A was added before or simultaneously as IFN-gamma. Natural IFN-alpha also exhibited similar inhibition and its suppressive effect was abolished in the presence of anti-IFN-alpha antibody. On the contrary, dexamethasone, a known inhibitor of class II MHC molecules on murine macrophages, showed a slight enhancing effect on class II MHC Ag. These results suggest an immunoregulatory role for IFN-alpha on non-lymphoid cells and that controlling elements for expression of class II MHC molecules may be different on various cell types as well as species.

Production of interleukin 1 by human endothelial cells.

Article

Apr 1986

Vascular endothelial cells (EC) play an important role in the emigration from the blood of the mononuclear cells that participate in the chronic inflammatory response. Because EC express a number of functions of cells of the monocyte/macrophage lineage, EC culture supernatants (ECSN) were examined for the presence of IL 1. In these supernatants, IL 1 activity was low when EC were cultured in the presence of serum. The low level of activity appeared to be due to the spontaneous production by the EC of inhibitors of the thymocyte proliferation assay of IL 1, of 70 kd and 9 kd, as measured by AcA Ultrogel filtration. When EC were cultured in the absence of serum, IL 1 activity was easily demonstrated in crude supernatants. Upon stimulation with LPS, the amounts of IL 1 activity were greatly increased. The release of IL 1 was an early event, detectable after 1 hr of incubation and reaching a maximum after 24 hr. The IL 1 activity produced by EC demonstrated a number of similarities to that of IL 1 produced by monocytes. On AcA 54 gel filtration, as with monocyte-derived IL 1, the IL 1 activity was found in two peaks of 50 to 60 kd and 16 to 18 kd. Upon chromatofocusing of the 16 to 18 kd peak, three active fractions were found, eluting near pH 7.0, 5.6, and 5.0. In addition, when LPS-stimulated ECSN and purified monocyte-derived IL 1 were incubated with a rabbit anti-IL 1 antibody, a parallel reduction in thymocyte-stimulating activity was observed, suggesting that the active agent in ECSN shared a common antigenic site with IL 1. The demonstration of IL 1 production by EC provides additional evidence that these cells, in addition to their functions as vascular cells, may also participate in some of the immune and nonimmune functions previously ascribed to macrophages.

Vascular endothelial cells and granulopoiesis: interleukin-1 stimulates release of G-CSF and GM-CSF

Article

Jan 1988

Cultured mononuclear phagocytes produce soluble factors that stimulate endothelial cells to release GM-colony-stimulating activity (GM-CSA). One such factor was recently identified as interleukin 1 (IL 1). Studies were designed to determine which types of granulopoietic factors are released by IL 1-stimulated endothelial cells. Supernatants from endothelial cells cultured for 3 days in medium containing IL 1 alpha and beta were tested in both murine and human CFU-GM colony growth assays. The effect of conditioned media on differentiation of WEHI-3B myelomonocytic leukemic cells was also examined. Control media containing IL 1 alone or unstimulated endothelial cell-conditioned media contained no detectable CSA in any bioassay. Medium conditioned by IL 1-stimulated endothelial cells stimulated the clonal growth of both human and murine CFU-GM and induced macrophage differentiation of WEHI-3B cells. Treatment of these conditioned media with a highly specific neutralizing monoclonal G-CSF antibody completely inhibited their activity in the murine CFU-GM assay, but only partially inhibited GM colony growth by human marrow. Treatment of the active conditioned media with a neutralizing rabbit anti-human GM-CSF antibody partially reduced the activity of the media in the human GM-colony growth assay. G-CSF radioimmunoassay of endothelial cell culture supernatants and Northern blot analysis of endothelial cell cytoplasmic RNA for GM-CSF gene transcripts confirmed that IL 1 induced expression of both G-CSF and GM-CSF genes. Because treatment of media with both antibodies abrogated all activity in the human GM colony growth assay, we conclude that IL 1-stimulated endothelial cells release both G and GM-CSF and that these are the only granulopoietic factors detectable in clonogenic assays released by these cells in vitro.

Endothelial activation: Its role in inflammatory and immune reactions

Article

Jan 1988

Recombinant tumor necrosis factor and immune interferon act singly and in combination to reorganize human vascular endothelial cell monolayers

Article

Jan 1986

Amer. J. Pathol

Article

Jan 1986

Lymphocyte Homing

Article

Apr 1989
ADV IMMUNOL

Lymphocyte recirculation serves to maximize the probability of a productive interaction between an antigen sequestered in a particular lymphoid organ and the typically rare lymphocytes that can respond to it. The efficiency of this process depends upon a massive and continuous procession of lymphocytes. In rat, for example, 4 X 107 lymphocytes per hour are returned to the blood via the thoracic duct, a major lymphatic vessel draining many of the secondary lymphoid organs. This rate is sufficient to replace the blood content of lymphocytes 10-20 times each day. The major portal of the entry of blood-borne lymphocytes into secondary lymphoid organs is through a specialized postcapillary venule, which is characterized by a cuboidal, rather than the typical flat, endothelial lining. An adhesive interaction of lymphocytes with these so-called high endothelial venules (HEV) initiates lymphocyte extravasation. The lymphocyte cell-surface receptors underlying HEV attachment are termed homing receptors, a nomenclature chosen to reflect the tendency of different lymphocyte populations to migrate to and extravasate within particular lymphoid organs. Thus, far homing receptors of three distinct HEV-binding specificities have been identified and more are likely to exist. The adhesive determinants on HEV, complementary to the lymphocyte homing receptors, are referred to in this review as HEV ligands. A brief description of the general characteristics of this cell-cell interaction; a review of the specificity thus far demonstrated or inferred for this interaction; the identity and nature of the homing receptors and their corresponding HEV associated ligands; and the possible role for specific carbohydrates as cell recognition determinants in lymphocyte-HEV binding are the four aspects of the lymphocyte-HEV interaction that this chapter discusses.

Endothelial cells augment T cell interleukin 2 production by a contact- dependent mechanism involving CD2/LFA-3 interaction

Article

May 1990

C. C. Hughes

We have demonstrated that endothelial cells (EC) augment IL-2 production by PHA-stimulated PBMC or purified CD4+ T cells and that the increase is apparent both in the amount of soluble IL-2 secreted and in the level of specific mRNA detectable by Northern blot hybridization. The ability of EC to affect levels of IL-2 cannot be reproduced by soluble factors, including the cytokines IL-1, IL-6, IFN-gamma, or TNF, conditioned medium from resting EC or IL-1, IFN-gamma- or TNF-treated EC, or from resting PBMC + EC cultures. Separation of the EC and PBMC by a Transwell membrane demonstrated that cell contact was required for augmentation of IL-2 synthesis and that this effect was unlikely to be mediated by a short-lived soluble signal. The cell-cell interaction required the ligand pair CD2/LFA-3, since augmentation could be inhibited by antibodies to these structures. Antibodies to ICAM-1, LFA-1, CD4, and MHC class II were without effect. A contact-dependent pathway involving CD2/LFA-3 interactions also may be used by EC to augment IL-2 production from T cells stimulated more specifically through the TCR/CD3 complex with antibody OKT3. This pathway provides a proliferative advantage to T cells stimulated with OKT3 in the presence of EC and may also be involved in the proliferative response of resting T cells to allogeneic class II MHC-expressing EC. We propose that EC augmentation of T cell IL-2 synthesis may be critical in the ability of EC to elicit primary T cell antigen responses and may have consequences for the development of localized cell-mediated immune reactions.

A homing receptor-IgG chimera as a probe for adhesive ligands of lymph node high endothelial venules

Article

Jun 1990

S. R. Watson

The binding of lymphocytes to high endothelial venules (HEV) within peripheral lymph nodes (pln) is thought to be mediated by a lectinlike adhesion molecule termed the pln homing receptor (pln HR). The cloning and sequencing of cDNAs encoding both murine and human pln HR revealed that these adhesion molecules contain protein motifs that are homologous to C-type or calcium dependent lectin domains as well as to epidermal growth factor (egf) and complement-regulatory protein domains. We have produced a novel, antibody-like form of the murine HR by joining the extracellular region of the receptor to a human IgG heavy chain. This antibody-like molecule is capable of recognizing carbohydrates, blocking the binding of lymphocytes to pln HEV, and serving as a histochemical reagent for the staining of pln HEV. This murine HR-IgG chimera should prove useful in analyzing the distribution of the HR ligand(s) in normal as well as in inflammatory states.

Naive and memory T cells show distinct pathways of lymphocyte recirculation

Article

Mar 1990

C. R. Mackay

In this report, we have addressed two questions concerning immunological memory: the way in which naive and memory T cells recirculate through the body, and the intrinsic rate of division within the naive and memory populations. We identified naive and memory T cells in sheep by their cell surface phenotype and their ability to respond to recall antigen. Memory T cells were CD2hi, CD58hi, CD44hi, CD11ahi, and CD45R-, as pertains in man. T cells that crossed from blood to the tissues of the hind leg and accumulated in the popliteal afferent lymph were all of memory phenotype. Conversely, T cells in efferent lymph, 90% of which entered the lymph node (LN) via high endothelial venules (HEV), were mostly of the naive phenotype (CD2lo, CD58lo, CD44lo, CD11alo, and CD45R+). The marked enrichment of these two phenotypes in different recirculatory compartments indicated that memory T cells selectively traffic from blood to peripheral tissues to LN (via afferent lymph), whereas naive T cells selectively traffic from blood to LN (via HEV). We argue that the differential use of these two recirculation pathways probably optimizes lymphocyte interactions with antigen. The nonrandom distribution of T cell subsets in various recirculatory compartments may be related to the relative proportion of memory cells in each subset. In particular, gamma/delta T cells in blood were almost exclusively of memory phenotype, and accumulated preferentially in afferent, but not in efferent, lymph. Finally, using the bromo-deoxyuridine labeling technique, we found that at least a sizeable proportion of memory T cells, whether in blood or afferent lymph, were a dividing population of cells, whereas naive T cells were a nondividing population. This result supports an alternative model of lymphocyte memory that assumes that maintenance of memory requires persistent antigenic stimulation.

Leukocyte interleukins induce cultured endothelial cells to produce a highly organized, glycosaminoglycan-rich pericellular matrix

Article

Nov 1984

Roberto Montesano

We report here that interleukins have a dramatic effect on extracellular matrix production by cultured endothelial cells. Human umbilical vein endothelial cells incubated with growth media conditioned by lectin-activated human peripheral blood mononuclear leukocytes undergo marked changes in cell shape and elaborate a highly organized extracellular material that is not detectable in untreated cultures. This material has the following characteristics: (a) it is not recognizable by electron microscopy unless the cationic dye, Alcian blue, is added to the fixative; (b) it is visualized as a network of branching and anastomosing fibrils of various thickness that can be resolved into bundles of fine filaments; (c) it is associated with the cell surface, extends between contiguous cells, and coats the culture substrate; (d) it is removed by digestion with glycosaminoglycan-degrading enzymes, such as crude heparinase and chondroitinase ABC. These results demonstrate that soluble factors released by activated peripheral blood mononuclear leukocytes (interleukins) stimulate cultured human umbilical vein endothelial cells to produce a highly structured pericellular matrix containing glycosaminoglycans (probably chondroitin sulfate and/or hyaluronic acid) as a major constituent. We speculate that this phenomenon corresponds to an early step of angiogenesis as observed in vivo as a consequence of interleukin release.

Direct demonstration of the lectin activity of gp90MEL, a lymphocyte homing receptor

Article

Sep 1990

Y Imai

Considerable evidence implicates gp90MEL as a lymphocyte homing receptor mediating lymphocyte attachment to high endothelial venules of lymph nodes in mouse. The protein appears to function as a calcium-dependent, lectin-like receptor as inferred primarily by the ability of specific carbohydrates to block its function and by the presence of a calcium-type lectin domain in its primary sequence. An ELISA assay is described which provides the first demonstration that the isolated protein has lectin activity and allows a further definition of its carbohydrate specificity. In addition to the monosaccharides mannose-6-phosphate and fructose-1-phosphate, ligand activity is shown for the sulfated glycolipid, sulfatide, and for two sulfated fucose-containing polysaccharides (fucoidin and egg jelly coat) from nonmammalian sources.

Inducible cell adhesion molecule 110 (INCAM-110) is an endothelial receptor for lymphocytes. A CD11/CD18-independent adhesion mechanism

Article

Apr 1990

G. E. Rice

Inducible cell adhesion molecule 110 (INCAM-110) is a 110-kD glycoprotein expressed on cytokine-activated human vascular endothelial cells. mAb blocking studies indicate that INCAM-110 and intercellular adhesion molecule 1 (ICAM-1) independently support the adhesion of lymphocytes to activated human umbilical vein endothelial cell monolayers. Anti-CD11a/CD18 antibodies with anti-INCAM-110 mAb E1/6 produce greater inhibition of lymphocyte adhesion than either reagent alone, suggesting that INCAM-110 and LFA-1 are not an obligate receptor-ligand pair. Blood monocytes, but not polymorphonuclear leukocytes, also appear to bind endothelial INCAM-110. Endothelial expression of INCAM-110 is upregulated at sites of inflammation, suggesting a role in the recruitment of mononuclear leukocytes.

Widespread and selective induction of major histocompatibility complex- determined antigens in vivo by gamma interferon

Article

Nov 1985

M. J. Skoskiewicz

gamma Interferon (IFN-gamma) caused remarkable increases in class I (H-2Kk) and class II (I-Ak) antigens throughout the body by 6-9 d. Heart, kidney, and adrenals showed increases of 4-8 times their previous levels of class I antigen content, while the pancreas and small intestine increased 13-17-fold. Lesser increases were found in spleen, liver, and lung, which showed higher resting antigenic potency. Increases of class II antigenicity of 6-10-fold were found in heart, kidney, pancreas, lung, liver, adrenal, and small intestine, with lesser increases in thymus and spleen, and none in lymph node. Topographical analysis revealed that IFN-gamma induced class I and II antigens on most tissues in a highly selective fashion. For example, the renal proximal tubules expressed large amounts of both class I and II antigens, whereas the distal tubules and collecting ducts did not. In some epithelial cells class I and II determinants were induced only on the basal aspects of the cell membrane. IFN-gamma caused a remarkable increase in class II-positive dendritic cells in the liver, pancreas, salivary glands, and thyroid. Whether these cells were of local or systemic origin is uncertain, but the finding of a simultaneous depletion of dendritic cells from lymph nodes and spleen raises the possibility that they may have been derived, at least in part, from these sites. The dynamic and selective induction of class I and II antigen expression by IFN-gamma is likely to be important in regulation of the immune response in tissues.

Transfer of experimental allergic encephalomyelitis to bone marrow chimeras. Endothelial cells are not a restricting element

Article

Dec 1987

D. J. Hinrichs

The adoptive transfer of clinical and histopathologic signs of experimental allergic encephalomyelitis (EAE) requires MHC compatibility between cell donor and cell recipient. The results of adoptive transfer studies using F1 to parent bone marrow chimeras as recipients of parental-derived BP-sensitive spleen cells indicate that this restriction is not expressed at the level of the endothelial cell but is confined to the cells of bone marrow derivation. Furthermore, these results indicate that the development of EAE is not dependent on the activity of MHC-restricted cytotoxic cells.

Phosphomannosyl receptors may participate in the adhesive interaction between lymphocytes and high endothelial venules

Article

Oct 1984

L M Stoolman

Normal and malignant lymphocytes can migrate from the bloodstream into lymph nodes and Peyer's patches. This process helps distribute normal lymphocytes throughout the lymphoid system and may provide a portal of entry for circulating malignant cells. An adhesive interaction between lymphocytes and the endothelium of postcapillary venules is the first step in the migratory process. We have recently shown that the simple sugars L-fucose and D-mannose, and an L-fucose-rich polysaccharide (fucoidin), can inhibit this adhesive interaction in vitro. We now report that mannose-6-phosphate, the structurally related sugar fructose-1-phosphate, and a phosphomannan, core polysaccharide from the yeast Hansenula holstii (PPME) are also potent inhibitors. Inhibitory activity was assessed by incubating freshly prepared suspensions of lymphocytes, containing the various additives, over air-dried, frozen sections of syngeneic lymph nodes at 7-10 degrees C. Sections were then evaluated in the light microscope for the binding of lymphocytes to postcapillary venules. Mannose-6-phosphate and fructose-1-phosphate were potent inhibitors of lymphocyte attachment (one-half maximal inhibition at 2-3 mM). Mannose-1-phosphate and fructose-6-phosphate had slight inhibitory activity, while glucose-1-phosphate, glucose-6-phosphate, galactose-1-phosphate, and galactose-6-phosphate had no significant activity (at 10 mM). In addition, the phosphomannan core polysaccharide was a potent inhibitor (one-half maximal inhibition at 10-20 micrograms/ml); dephosphorylation with alkaline phosphatase resulted in loss of its inhibitory activity. Preincubation of the lymphocytes, but not the lymph node frozen sections, with PPME resulted in persistent inhibition of binding. Neither the monosaccharides nor the polysaccharide suppressed protein synthesis nor decreased the viability of the lymphocytes. Furthermore, inhibitory activity did not correlate with an increase in negative charge on the lymphocyte surface (as measured by cellular electrophoresis). These data suggest that a carbohydrate-binding molecule on the lymphocyte surface, with specificity for mannose-phosphates and structurally related carbohydrates, may be involved in the adhesive interaction mediating lymphocyte recirculation.

Three distinct classes of regulatory cytokines control endothelial cell MHC antigen expression. Interactions with immune gamma interferon differentiate the effects of tumor necrosis factor and lymphotoxin from those of leukocyte alpha and fibroblast beta interferons

Article

Mar 1988

L A Lapierre

Recombinant preparations of TNF and lymphotoxin (LT) increase the expression of class I MHC antigens on cultured human endothelial cells (EC) without inducing expression of class II antigens. These actions are similar to those of rIFN-alpha or rIFN-beta. However, TNF and LT differ from IFN-alpha/beta in that the former synergize with IFN-gamma for class I regulation whereas the latter do not. Furthermore, LT or TNF do not affect IFN-gamma-mediated class II induction at optimal class I inducing concentrations (100 U/ml), whereas IFN-alpha and IFN-beta (at their optimal concentrations of 1,000 U/ml) are strikingly inhibitory. LT and TNF also can further increase expression of class I antigens on cells already maximally stimulated by IFN-alpha or IFN-beta. A recombinant preparation of IL-6 (formerly called 26-kD protein, IFN-beta 2, or B cell stimulating factor 2) was without effect on class I expression in EC. These data make it seem unlikely that the actions of LT or TNF on EC expression of MHC antigens are mediated through autocrine or paracrine production of IFN-alpha, IFN-beta or IL-6. More importantly, they suggest that LT or TNF are more likely to be immunostimulatory, whereas IFN-alpha or IFN-beta are more likely to be immunoinhibitory in vivo, a consideration of potential relevance for cytokine administration to various patient populations.

Ia expression by vascular endothelium is inducible by activated T cells and by human gamma interferon

Article

Apr 1983

J S Pober

We have used monoclonal antibody binding, measured by radioimmunoassay, fluorescence flow cytometry, and ultrastructural immunocytochemistry, to measure expression of Ia antigens on cultured human umbilical vein endothelial (HUVE) cells. Under standard culture conditions, HUVE cells do not express Ia antigens. However, treatment of primary HUVE cultures with phytohemagglutinin induces the expression of Ia antigens. Every endothelial cell in the culture becomes Ia-positive and endothelial cells appear to synthesize Ia. HLA-A,B is concomitantly increased. The expression of Ia appears to be mediated by T cells because (a) pretreatment of primary HUVE cultures with OKT3 plus complement blocks the action of the lectins but not of medium conditioned by lectin-activated peripheral blood mononuclear cells; (b) co-culture of endothelial cells with allogeneic T cells, in the absence of lectin, also induces endothelial Ia; and (c) human immune (gamma) interferon, produced by Chinese hamster ovary cells transfected with the human gamma interferon gene, directly induces endothelial Ia. During co-culture with lymphocytes, about one-third of the endothelial cells are Ia-positive after 24 h and all of the endothelial cells are Ia-positive by 72 h. Proliferation of allogeneic T cells starts by 96 h and peaks at 144 h. Thus, endothelial Ia appears sufficiently early to be a determinant for the proliferation of allogeneic T cells. Inducible expression of Ia by endothelium may be important both for allograft rejection and for recruitment of circulating T cells into the site of an immune response.

Induction and detection of a human endothelial activation antigen in vivo

Article

Aug 1986

R S Cotran

We used a murine mAb, H4/18, raised by immunization with IL-1-treated human umbilical vein endothelial cell cultures, to localize an endothelial activation antigen in induced human delayed hypersensitivity reactions (DHR) and in pathological tissues. We used streptococcus varidase to elicit DHR in human skin and we examined sequential skin biopsies with the immunoperoxidase technique. There was no staining for H4/18 binding antigen in normal endothelium of skin and other tissues; strong positive staining, localized to vascular endothelium, was seen at 16 and 23 h but disappeared by 6 d, when the DHR had faded. H4/18 binding antigen, also confined to endothelium, was detected in lymph nodes, skin, and other tissues exhibiting immune/inflammatory reactions. The studies indicate that H4/18 is a useful marker for activated endothelium in vivo and they support the relevance of in vitro studies on inducible endothelial cell functions.

Regulated expression and binding of three VLA (S1) integrin receptors on T-cells

Article

May 1990
NATURE

Regulated adhesion of T cells to extracellular matrix (ECM) proteins is likely to be essential in T cell migration. Constitutive binding of various other cell types to ECM components is mediated by members of the VLA (very late antigen) subfamily of integrins. We describe here the regulated binding of resting CD4+ human T cells to ECM through three VLA integrins: VLA-4 and VLA-5 binding to fibronectin (FN), and a novel pathway of VLA-6 binding to laminin (LN). Binding to ECM is regulated in two ways. First, unlike other VLA-mediated interactions, VLA binding activity of the T cells is rapidly and dramatically augmented with cell activation without change in level of expression of the VLA molecules. Second, binding is regulated with T-cell differentiation; memory T cells express three- to four-fold more VLA-4, VLA-5, and VLA-6 than do naive cells, and bind more efficiently through them to FN and LN.

Leu-8/TQ1 is the human equivalent of the Mel-14 lymph node homing receptor

Article

Nov 1989
NATURE

THE human pan-leukocyte antigen Leu-8 has attracted wide interest because its presence or absence identifies suppressor-inducer and helper-inducer CD4+ T-lymphocyte subsets respectively. We report here that Leu-8 is the human homologue of the mouse Mel-14 homing receptor, a molecule that promotes the initial adhesion of blood-borne lymphocytes to the specialized post-capillary endothelium of peripheral lymph nodes. We also show that Leu-8 can adopt both conventional and phospholipid anchored forms, a finding that may have relevance in the context of antigen shedding following activation or homing. The assignment of lymphocytes to different functional classes based on lymph node homing potential may represent a more general association between lymphocyte function and tissue distribution.

Gamma-interferon transcriptionally regulates an early-response gene containing homology to platelet proteins

Article

Jun 1985
NATURE

Interferons are a family of proteins first identified by their ability to induce cellular resistance to infection by many viruses. In addition to the antiviral properties it shares with the alpha- and beta-interferons, gamma-interferon (IFN-gamma), a lymphokine secreted by activated T cells, activates macrophages, stimulates B cells, increases fibroblast and endothelial cell resistance to many non-viral intracellular parasites and modulates cell-surface proteins central to immune cell regulation1-13. To identify molecules involved in the IFN-gamma response and characterize their modulation, we have isolated genes that are induced following recombinant IFN-gamma treatment of U937 cells, a histiocytic lymphoma cell line with monocytic characteristics14,15. We report here the molecular cloning and characterization of a gene regulated by rIFN-gamma in U937 cells as well as in human mononuclear cells, fibroblasts and endothelial cells. Messenger RNA from this gene is induced within 30 min of rIFN-gamma treatment and demonstrates maximal (>30-fold) accumulation within 5 h. Increased transcription is partly responsible for this accumulation. This gene encodes a protein of relative molecular mass (Mr) 12,378 which has significant amino-acid homology to platelet factor-4 and beta-thromboglobulin, two chemo-tatic proteins released by platelets on degranulation. This IFN-gamma-inducible protein may be a member of a family of proteins involved in the inflammatory process.

Lymphocyte recognize human vascular endothelial and dermal fibroblast Ia antigens induced by recombinant immune interferon

Article

Oct 1983

T-lymphocyte-mediated responses to the cellular components of blood vessels are important in rejection of allografts1–3. The induction of cytolytic T lymphocytes (CTLs) depends on recognition of foreign class II major histocompatibility complex antigens (human HLA-DR, DC/DS, SB and others, collectively referred to as Ia) on the target cells whereas killing by CTLs usually depends on recognition of foreign class I antigens (HLA-A,B)4, although some alloreactive CTLs recognize foreign Ia instead of HLA-A, B (refs 5–8). The expression of Ia antigens has traditionally been regarded as restricted to immunological cell types, and the presence of class II antigen-bearing ‘passenger’ leukocytes in rodent organ grafts appears necessary for graft rejection9–11. Recently, Ia antigens have been observed by immunofluorescence microscopy on human renal and dermal capillary endothelium12–15. We have previously shown that human umbilical vein endothelial (HUVE) cells in standard culture conditions do not bear Ia antigens, but may be induced to do so by products of lectin- or alloantigen-activated T lymphocytes16,17. Furthermore, we found that recombinant immune interferon (IFN-γ), free of other lymphokines, is a potent inducer of Ia expression in HUVE cells17. Here we report that IFN-γ also induces Ia expression on human foreskin capillary endothelial (HFCE) cells, HUVE cells transformed by Simian virus 40 viral DNA (SV-HUVE cells) and human dermal fibroblast (HDF) cells in culture. Further, we present evidence that Ia present on HUVE cells and HDF cells can be functionally recognized by human T cells, resulting in a two-way interaction between T cells and mesenchymal cells that may be important in allograft rejection.

Vascular endothelial cells in cell‐mediated immunity: Adoptive transfer with in vitro conditioned cells is genetically restricted at the endothelial cell barrier

Article

Jan 1985
J CELL BIOCHEM

Delayed-type hypersensitivity (DTH) is a cell-mediated immune response that can be adoptively transferred in rats when greater than 2 × 108 cells from peritoneal exudate, lymph nodes, or spleen are used. We have shown that by using an in vitro conditioning step with antigen, transfer can be subsequently carried out with as few as 2 × 107 spleen cells. The magnitude of DTH was reflected in ear swelling after intradermal injection of antigen [tuberculin or keyhole limpet hemocyanin (KLH)] and confirmed histologically. The transfer was antigen specific, requiring the sensitizing antigen in both the in vitro conditioning step and in the ear test challenge. Adoptive transfer with conditioned cells was genetically restricted by alleles of the RT-1 region [major histocompatibility complex (MHC) of the rat]. Brown Norway strain (n haplotype) immune cells would not transfer DTH to Lewis (1 haplotype), ACI (a haplotype), or Buffalo (b haplotype) rats, whereas each strain would transfer DTH to syngeneic recipients. Moreover, this pattern of restriction held for all strains when tested in reciprocal fashion. In additional experiments, F1 to parental bone marrow chimeras were constructed so that bone-marrow-derived cells and non-bone-marrow-derived cells were of different RT-1 haplotypes. When these chimeras were used as recipients, transfer of DTH was only observed when immune donor cells and recipient non-bone-marrow-derived cells were syngeneic. These results point to the critical role of non-bone-marrow-derived cells (endothelial cells) in the DTH reaction.

HLA‐D Region Products Are Expressed in Endothelial Cells Detection by Primed Lymphocyte Typing

Article

Feb 1980
TISSUE ANTIGENS

The mixed lymphocyte endothelial cell culture was studied by the primed lymphocyte typing (PLT) technique. By comparing the HLA-D/DR specificity of the secondary response when using either peripheral blood mononuclear cells (PBM) or endothelial cells from umbilical cords for priming or restimulation of lymphocytes, it was found that PBM from newborns would induce a clear-cut specificity for HLA- D/DR when used for priming as well as for restimulation. No HLA-D/DR specificity was seen, however, when endothelial cells were used for restimulation of lymphocytes primed to HLA- D/DR on PBM. On the other hand, lymphocytes primed to endothelial cells showed significant, albeit not very strong specificity for HLA-D/DR when restimulated with PBM. Our experiments suggest that HLA-D region products are present on endothelial cells, and thus confirm and extend serological studies using anti DR antisera.

Functional and biosynthetic changes in endothelial cells of vessels in chronically inflamed tissues: Evidence for endothelial control of lymphocyte entry into diseased tissues

Article

Jul 1988
J PATHOL

Anthony J Freemont

In vitro lymphocyte adhesion to, and selective radiosulphate uptake by, endothelial cells has been demonstrated in chronically inflamed tissues of patients with peptic ulceration, rheumatoid disease, pilonidal sinus, autoimmune thyroiditis, polymyositis, primary biliary cirrhosis, and pyelonephritis. These characteristics have been described previously in endothelial cells functionally specialized for promoting lymphocyte traffic from blood to lymph node parenchyma. It is suggested that these observations indicate that some vessels in inflamed tissues may be, at least in part, responsible for the selective accumulation of lymphocytes within the tissue. Manipulating the development of this type of vessel may offer a novel way of influencing the progress of inflammatory disorders.

An immunohistochemical study on phenotypic heterogeneity of human pulmonary vascular endothelial cells

Article

Sep 1988

The existence of a subpopulation of human vascular endothelial cells (EC) has been demonstrated in the liver with the aid of immunohistochemical techniques. In this study, we investigated the antigenic and functional properties of the vascular EC in human lung. Alveolar capillary EC shared antigens with a peripheral blood monocyte/ macrophage subset capable of presenting soluble antigens and triggering autologous mixed lymphocyte reactions. That is to say that the alveolar capillary EC were HLA-DR+, OKM1–, and OKM5+. In addition, these EC frequently expressed interleukin-1. These facts suggest that alveolar capillary EC may play an important role in immunological responses in the lung. The antigens were, however, absent or only faintly visible on the vascular EC of medium and small vessels. In contrast, Factor VIII/von Willebrand factor antigen (FVIIIRAg), which is produced in vascular EC was heavily stained in the EC of medium and small vessels, but only weakly stained in the alveolar capillary EC. These immunohistochemical findings suggest that in different anatomical compartments in the lung vascular EC express phenotypic properties heterogeneously. They may play differing biological roles or serve different immunological functions in normal and pathological states in the lung.

Class II major histocompatibility antigen expression on coronary arterial endothelium in a patient with Kawasaki disease

Article

Mar 1990
Hum Pathol

To investigate the class II major histocompatibility antigen expression on coronary arterial endothelium of Kawasaki disease and immunophenotypes of the infiltrating cells in the coronary vascular lesions, myocardial sections from a patient who died during the acute stage of Kawasaki disease were studied using an immunoperoxidase technique. The mononuclear cells in the lesions mainly consisted of macrophages and T cells, whereas B cells and cells were not seen. The majority of T cells reacted with Leu-3a antibodies, and only a few reacted with Leu-2a antibodies. Cells bearing the interleukin-2 receptor, indicative of activated T cells, were also found in the lesions. To determine the distribution of class II antigen, we used anti-HLA-DR antibodies. The massive expression of HLA-DR antigen on mononuclear cells was found in the lesions. In addition, the HLA-DR activation antigen was expressed on the coronary arterial endothelium at the infiltrates in which macrophages and T cells coexisted. In contrast, coronary arterial endothelium did not express HLA-DR antigens in the myocardial tissues of controls (n = 4). HLA-DR+ endothelial cells may play an important role in the development of Kawasaki vasculitis.

Human naive and memory T cells: reinterpretation of helper-inducer and suppressor-inducer subsets

Article

Jan 1988
Immunol Today

The subsets of human peripheral blood T cells identified by CD45R antibodies (such as 2H4) and by CDw29 antibodies (such as 4B4) are assuming increasing importance in studies of both basic immunology and clinical medicine. Here, Martin Sanders and colleagues propose that these subsets represent naive and memory (previously activated) T cells, respectively, and discuss the current understanding of these subsets in the light of this reinterpretation.

Stimulation of endothelial cell binding of lymphocytes by Tumor Necrosis Factor

Article

Sep 1987

Lymphokines and monokines have been reported to affect endothelial cell (EC) morphology and function. In experiments here described, we have demonstrated that recombinant tumor necrosis factor (TNF) stimulates the adhesion of T lymphocytes to confluent monolayers of human umbilical vein EC. The increase in adhesion induced by TNF was EC-specific inasmuch as preincubation of the lymphocytes with TNF did not alter binding, and preincubation of human dermal fibroblasts with TNF did not increase their inherently low adhesiveness for lymphocytes. Stimulation of T-EC binding occurred after treatment of the EC with as little as 0.01 U/ml (1 pg/ml) of TNF. In kinetic experiments, preincubation of EC with TNF for 4 hr resulted in optimal adhesion. TNF-treated EC retained their increased adhesiveness after fixation with paraformaldehyde, suggesting that TNF stimulated binding by increasing the expression or accessibility of EC surface receptors for lymphocytes. Although antibodies to the lymphocyte function-associated antigen 1 alpha- or beta-chains on the T cell markedly inhibited unstimulated T-EC binding, such antibodies had no effect on the increase in EC adhesiveness induced by TNF, indicating that the increased binding resulted from the generation of an alternate binding receptor on the EC membrane. These findings provide additional evidence that cytokines participate in the mobilization of mononuclear cells in the chronic inflammatory reaction by stimulation of the adhesiveness of endothelium for circulating lymphocytes.

The human mixed lymphocyte-endothelium culture reaction

Article

Jul 1975

Cells separated from the wall of the umbilical cord vein by collagenase digestion could be identified as endothelial by their characteristic ultrastructure, their growth pattern in culture, and their microscopical morphology. These cells, both freshly explanted and after long-term culturing, were capable of stimulating allogeneic lymphocytes in vitro. Control experiments indicated that this stimulation was not attributable to contamination of the endothelial cell suspensions by foetal fibroblasts or passenger lymphocytes. The dose response characteristics and kinetics of the lymphoproliferative response using endothelial stimulating cells was similar to mixed lymphocyte cultures. Sera which were capable of inhibiting the mixed lymphocyte culture response were relatively ineffective in inhibiting the stimulation caused by endothelial cells.

Human Ia-like antigen in non-lymphoid organs

Article

Nov 1979

Human Ia-like antigens in liver and kidney were shown by the immunofluorescence assay to be present mostly in the endothelial-mesenchymal cells of these organs. The parenchymal cells apparently contained no human Ia-like antigens. The antigens in liver and kidney were purified and shown to have the same subunit structure as human Ia-like antigens of cultured B-lymphoid cells. The human Ia-like antigens in non-lymphoid organs, not only in liver and kidney but also in testis, heart, muscle and brain, carried all the xenoantigenic characteristics of human Ia-like antigens expressed on lymphoid cells of B-cell lineage.

Endothelial cell Lymphocyte Function-Associated Antigen-3 and an unidentified ligand act in concert to provide costimulation to human peripheral blood CD4+ T cells

Article

Nov 1991

Our previous studies have demonstrated that cultured human endothelial cells (EC) provide costimulation to PHA-activated CD4+ T cells, measured as augmentation of IL-2 synthesis, through a cell contact-department pathway. Here we show that fixed and living EC provide comparable degrees of costimulation to CD4+ T cell populations, indicating that EC costimulation does not depend upon active metabolism. EC achieve these effects in part by utilizing lymphocyte function-associated antigen-3 (LFA-3) to interact with T cell CD2 as shown by observations that EC augmentation of IL-2 is partially (50-70%) blocked by eight of eight mAb tested which recognize LFA-3; that purified phosphatidylinositol-linked LFA-3 (PI-LFA-3) can also provide costimulation to CD4+ T cells; and that there is a delay of the EC effect on CD4+ T cells which express low levels of CD2 compared to those which express high levels of CD2. However, three lines of evidence suggest that EC also utilize at least one additional ligand. First, there is incomplete replacement of the EC effect by PI-LFA-3 such that the costimulatory ability of EC combined with PI-LFA-3 is additive at all concentrations of PI-LFA-3 tested. Second, costimulation by PI-LFA-3, but not by EC, is fully inhibited by anti-CD2 or anti-LFA-3 mAb. Finally, costimulation by PI-LFA-3, but not by EC, is completely suppressed by cyclosporine A. We have not formally identified the second ligand but it does not appear to be intercellular adhesion molecule-1, vascular cell adhesion molecule-1, CD44, or B7/BB1.

VCAM-1 on activated endothelium interacts with the leukocyte integrin VLA-4 at a site distinct from the VLA-4/Fibronectin binding site

Article

Mar 1990
CELL

Cytokine-activated human endothelial cells express vascular cell adhesion molecule-1 (VCAM-1), which binds lymphocytes. We now identify the integrin VLA-4 as a receptor for VCAM-1 because VLA-4 surface expression on K-562 cells (following transfection of the VLA alpha 4 subunit cDNA) resulted in specific cell adhesion to VCAM-1, and anti-VLA-4 antibodies completely inhibited VCAM-1-dependent cell-cell attachment. In addition, VLA-4 expression allowed K-562 cells to attach to the heparin II binding region (FN-40) of fibronectin. However, VLA-4/VCAM-1 and VLA-4/FN-40 interactions are readily distinguishable: only the former was inhibited by the anti-VLA-4 monoclonal antibody HP1/3, and only the latter was inhibited by soluble FN-40. The VCAM-1/VLA-4 ligand-receptor pair may play a major role in the recruitment of mononuclear leukocytes to inflammatory sites in vivo.

Expression of the human leukocyte adhesion molecule, LAM1. Identity with the TQ1 and Leu-8 differentiation antigens

Article

Feb 1990

The LAM1 molecule is a member of the new family of cellular adhesion/homing molecules that contain a lectin-like domain at their amino-terminal end followed by an epidermal growth factor-like domain and short consensus repeat units like those found in C3/C4 binding proteins. Two mAb that react with the leukocyte adhesion molecule 1 (LAM1) were produced and used to examine the cell-surface expression of LAM1. The anti-LAM1 antibodies were reactive with the majority of blood lymphocytes, NK cells, neutrophils, and monocytes. LAM1 was also expressed by subpopulations of phenotypically immature and mature thymocytes. Blood lymphocytes rapidly modulated LAM1 from the cell surface during PMA exposure for 60 min. Coordinate with the loss of LAM1 from the cell surface, PMA-treated lymphocytes lost the ability to bind to lymph node high endothelial venules, indicating that expression of LAM1 may play a role in lymphocyte homing. Mitogen stimulation of blood T and B lymphocytes also resulted in decreased LAM1 expression, but at a slower rate. LAM1 was only weakly expressed by a minority of spleen lymphocytes. However, culturing spleen lymphocytes in media alone resulted in increased expression of LAM1 by a subpopulation of the cells (40 to 60%). Concomitant mitogen stimulation of spleen lymphocytes resulted initially in down-regulation of LAM1 expression followed by increased expression of LAM1 and then subsequent loss of LAM1 from the cell surface. The pattern of anti-LAM1 antibody reactivity was identical to that reported for the TQ1 and Leu-8 antibodies, and all of these antibodies reacted with cells transfected with the LAM1 cDNA. Thus, LAM1 is broadly expressed by leukocytes, and binding of LAM1 may participate in the process of leukocyte extravasation into lymphoid organs or sites of acute inflammation with subsequent loss of LAM1 from the cell surface.

Increased lymphocyte adherence to human arterial endothelial cell monolayers in the context of allorecognition

Article

May 1990

The interactions of alloreactive T lymphocytes with the vascular endothelium were studied in an in vitro model of lymphocyte adherence to cultured human arterial endothelial cell (HAEC) monolayers. Donor-primed lymphocytes (DPL) were shown to have significantly greater adherence to donor HAEC than were third-party primed lymphocytes. Limiting dilution analysis of adherent DPL showed an enrichment of donor-reactive lymphocytes compared with nonadherent DPL. This study examines the allospecific nature of this increased lymphocyte adherence. HAEC constitutively express class I HLA Ag and can be induced by IFN-gamma to express class II Ag. DPL adherence to class I+ HAEC was inhibited only in the presence of mAb directed against class I Ag. DPL adherence to class I+ and class II+ HAEC was inhibited in the presence of mAb directed against class I and class II Ag. Class I- and class II-specific adherence was also shown to involve CD8 and CD4 molecules, respectively, whereas lymphocyte function-associated Ag do not appear to play a major role in long term alloreactive lymphocyte adherence to HAEC. These findings suggest that alloreactive lymphocyte adherence to HAEC is mediated by two mechanisms. One is based on allorecognition, primarily of HLA Ag, and the other is related to presumably non-Ag-specific interactions between activated lymphocytes and the vascular endothelium. The studies presented provide evidence to suggest that HLA-specific lymphocyte adherence to endothelium may significantly contribute to the development of alloreactive lymphocyte infiltrates within the allograft.

IL-4 regulates endothelial cell activation by IL-1, tumor necrosis factor, or IFN-gamma

Article

Sep 1990

Alteration in the surface membrane of endothelial cells (EC) is a feature of endothelial activation both at sites of inflammation in vivo and after stimulation with cytokines in vitro. The effects of stimulating EC with IL-1 or TNF include enhanced adhesiveness for polymorphonuclear leukocytes (PMN) and T cells, the induction of EC leukocyte adhesion molecule-1 (ELAM-1) expression, and the increased expression of intercellular adhesion molecule-1 (ICAM-1) and the 1.4C3 Ag. In contrast, IFN-gamma stimulation increases EC binding of T cells but not PMN and enhances ICAM-1 expression but not ELAM-1 or 1.4C3 Ag expression. Recently we have reported that the T cell-derived cytokine IL-4 also increases EC adhesiveness for T cells but not PMN. In this study we have examined the effect of IL-4 on the expression of several cytokine-inducible EC activation Ag, by using a previously described ELISA technique. IL-4 modulation of activation Ag expression was concentration dependent, optimal at around 100 U/ml, and exhibited a unique pattern compared to that seen with the other cytokines. Although, IL-4 stimulation increased 1.4C3 Ag expression (p less than 0.001), it significantly inhibited constitutive ICAM-1 expression (p less than 0.01) and did not induce ELAM-1. Furthermore, IL-4 exhibited significant synergy with IL-1 or TNF in inducing 1.4C3 Ag expression (p less than 0.001) but inhibited the increased expression of ICAM-1 produced by IL-1, TNF, or IFN-gamma (p less than 0.01) and inhibited the induction of ELAM-1 by IL-1 and TNF (p less than 0.001). In contrast, IL-4 had no effect on the expression of EC HLA-class I, -DR, -DP, or -DQ and neither enhanced nor inhibited the effect of IFN-gamma on the expression of these molecules. Finally, although IL-4 alone caused little if any shape change in EC monolayers, it strongly synergized with TNF or IFN-gamma in causing a change in shape to a more fibroblastic morphology. These observations indicate that IL-4 increases EC adhesiveness for T cells by the induction of a different adhesion molecule to ICAM-1. Furthermore, the ability of IL-4 to both enhance and inhibit the expression of activation Ag on EC already activated by IL-1, TNF, or IFN-gamma suggests that it may be important in altering the quality of inflammatory responses such as may occur during the development and maintenance of chronic or immune-mediated inflammation.

T cells bind to cytokine-activated endothelial cells via a novel, inducible sialoglycoprotein and endothelial leukocyte adhesion molecule-1

Article

Sep 1990

The action of human rIL-1 beta on confluent, quiescent monolayers of human umbilical vein endothelial cells (HUVEC) has been studied for the induction of new membrane proteins. Two approaches have been taken. The first is a quantitative two-dimensional gel analysis of [35S]cysteine-labeled membrane proteins of HUVEC with and without cytokine treatment. This analysis indicates that there are a restricted number of new membrane proteins synthesized in the first 6 h of IL-1 treatment, on the order of 19 out of a total of over 600 detectable proteins. Second, we have prepared two mAb (1E7 and 2G7) to different epitopes of a major inducible sialoglycoprotein with molecular mass of 114 kDa and an isoelectric point of 4.6 to 4.8. These antibodies were compared with two additional antibodies, 3B7 and 7A9, which were shown to react with the endothelial leukocyte adhesion molecule-1 (ELAM-1) protein as expressed in COS cells. The 1E7/2G7 protein is distinct from ELAM-1, based upon biochemical comparisons as well as the inability of the 1E7 and 2G7 antibodies to react with ELAM-1-transfected COS cells. The protein defined as 1E7/2G7 is neither expressed constitutively nor in an inducible manner on PBMC, granulocytes, platelets, fibroblasts, or keratinocytes. The 7A9 and 3B7 antibodies are shown to block granulocyte binding to IL-1-activated HUVEC. The 2G7 antibody is effective at inhibiting the binding of T cells but not granulocytes to IL-1-activated endothelium, suggesting this new protein is an adhesion protein that may be active in vivo in T cell-endothelial cell adhesion-related events such as inflammation or lymphocyte recirculation. In addition, T cells were shown to utilize the ELAM-1 protein in binding to cytokine-activated HUVEC. Antibodies directed to both proteins had additive effects on inhibition of T cell adhesion.

IFN-?? enhances endothelial activation induced by tumor necrosis I factor but not IL-1

Article

Oct 1990

Previous studies have established that different cytokines induce distinct patterns of activation in cultured endothelial cells (EC). Treatment of EC with either TNF or IL-1 causes transient induction of endothelial leukocyte adhesion molecule-1 (ELAM-1) and a sustained increase in intercellular adhesion molecule-1 (ICAM-1) expression. TNF but not IL-1 also increases class I MHC Ag expression. IFN-gamma, which by itself increases EC class I MHC and ICAM-1 but does not induce ELAM-1 expression, has been found to act synergistically with TNF to increase class I expression. In our study, we have further examined IFN-gamma effects on both TNF and IL-1 beta responses. In contrast to IFN-gamma plus TNF cotreatment, IFN-gamma up-regulation of class I MHC molecules is not augmented by cotreatment with IL-1 beta. IFN-gamma plus TNF cotreatment synergistically increases ICAM-1 expression by 24 h of cotreatment, whereas IFN-gamma plus IL-1 beta cotreated EC show at most additive increases. IFN-gamma increases TNF-induced ELAM-1 expression such that a greater number of EC express ELAM-1 at both 4 and 24 h in the presence of IFN-gamma plus TNF compared to cultures treated with TNF alone, although the maximal level of surface expression on individual cells is not increased. Inasmuch as these times represent pre- and post-peak expression time (6 h), respectively, IFN-gamma appears both to accelerate and to prolong transient ELAM-1 expression. Although similar interactions are seen with IL-1 beta, IFN-gamma has a consistently greater effect on TNF-induced compared to IL-1-induced ELAM-1 expression. We next explored the possible mechanism(s) of the synergy between IFN-gamma and TNF. IFN-gamma and TNF do not cooperatively enhance total protein synthesis. Unexpectedly, IFN-gamma and IL-1 beta combine to depress protein synthesis, which may contribute to the failure of these cytokines to positively interact. IFN-gamma and TNF cooperatively increase ELAM-1 mRNA at 6 h and ICAM-1 mRNA at both 6 and 24 h, whereas IFN-gamma and IL-1 show markedly less cooperative augmentation of these transcripts. We conclude that IFN-gamma enhances TNF-induced EC activation by selectively and synergistically increasing synthesis of specific surface molecules, whereas IL-1-induced EC activation is largely unaffected by IFN-gamma.

IFN-γ regulates the expression of the adhesion molecule ELAM-1 and IL-6 production by human endothelial cells

Article

Nov 1990

In this study two new in vitro effects of IFN-gamma on human umbilical vein endothelial (HUVE) cells were described. First, it was shown that the expression of the adhesion molecule ELAM-1 on activated HUVE cells can be modulated by IFN-gamma. ELAM-1 is normally not expressed by HUVE cells, but its expression can rapidly be induced by TNF, IL-1, or LPS. Maximal expression is reached after 4 to 6 h of activation, and after 24 h the expression disappeared. Whereas IFN-gamma per se did not induce expression of ELAM-1, it enhanced and prolonged the expression of ELAM-1. This enhancement occurred when IFN-gamma was added before activation as well as when added simultaneously with activation. When IFN-gamma was added 6 or 9 h after the activation, the normally ongoing reduction of expression was not only retarded, but the expression increased for at least 3 h. Moreover, IFN-gamma abrogated the refractory period for restimulation. Neither IFN-beta nor IL-6 had any effect on the expression of ELAM-1. The second effect of IFN-gamma on HUVE cells is the capacity to enhance the IL-6 production by these cells. Prestimulation as well as coincubation of IFN-gamma with TNF, IL-1, or LPS resulted in a strongly augmented production of IL-6. The effects of IFN-gamma may in vivo play a role in the regulation of an inflammatory reaction, because ELAM-1 is an adhesion molecule for neutrophils, and IL-6 has an enhancing effect on the cytotoxicity of neutrophils.

Immunologic Interactions of T Lymphocytes with Vascular Endothelium

Abstract

No full-text available

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