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Reversal of thymic atrophy
Abstract
Age-associated thymic atrophy is a key event preceding the inefficient functioning of the immune system, resulting in a diminished capacity to generate new T-cells. This thymic involution has been proposed to be due to changes in the thymic microenvironment resulting in its failure to support thymopoiesis. A key cytokine in the early stages of thymocyte development is IL-7 and expression levels are greatly reduced with age. The ability of IL-7 to restore the immune system by enhancing thymic output remains controversial. In this review, we highlight the advances in molecular approaches used to evaluate recent thymic emigrants and assess the success of these strategies in determining whether IL-7 can lead to immune reconstitution.
... Histologic changes include alteration of normal architecture, enlarged perivascular spaces, leading to decreased thymic epithelial space, reduced cortical thickness, and loss of corticomedullary demarcation. 98,225 There is diffuse hypocellularity, with overall reduction in lymphocytic cell numbers, particularly within the cortex, with relative sparing of the medulla. There may be increased numbers of perivascular B cells and plasma cells, particularly at the corticomedullary junction, and formation of lymphoid follicles with germinal centers. ...
... Although the thymus is necessary for the production of new naïve T cells in the neonate and juvenile, it has been demonstrated that the adult thymus is also capable of T cell reconstitution, albeit at a reduced capacity. 11,98,108 Aging is associated with a reduction in the contribution of the thymus to the naïve T-cell pool, but with no significant decline in the total number of T cells in the peripheral T-cell pool. 11,103,235 Many non-infectious and infectious disease states are accompanied by thymic changes (Figure 2). ...
The development of a protocol to reproducibly induce thymic atrophy, as occurs in feline immunodeficiency virus (FIV) infection and other immunosuppressive diseases, and to consistently estimate thymic volume, provides a valuable tool in the search of innovative and novel therapeutic strategies. Magnetic resonance imaging (MRI) using the short tau inversion recovery (STIR) technique, with fat suppression properties, was determined to provide an optimized means of locating, defining, and quantitatively estimating thymus volume in young cats. Thymic atrophy was induced in four, 8-10-week-old kittens with a single, directed 500 cGy dose of 6 MV X-rays from a clinical linear accelerator, and sequential MR images of the cranial mediastinum were collected at 2, 7, 14, and 21 days post irradiation (PI). Irradiation induced a severe reduction in thymic volume, which was decreased, on average, to 47% that of normal, by 7 days PI. Histopathology confirmed marked, diffuse thymic atrophy, characterized by reduced thymic volume, decreased overall cellularity, increased apoptosis, histiocytosis, and reduced distinction of the corticomedullary junction, comparable to that seen in acute FIV infection. Beginning on day 7 PI, thymic volumes rebounded slightly and continued to increase over the following 14 days, regaining 3-35% of original volume. These findings demonstrate the feasibility and advantages of using this non-invasive, in vivo imaging technique to measure and evaluate changes in thymic volume in physiologic and experimental situations. All experimental protocols in this study were approved by the Institutional Animal Care and Use Committee (IACUC) at Auburn University.
... After regression with age, thymopoiesis is reduced and thymic function decreases and replaced with adipose tissue [8,19,29,34,37]. Thymic function has been evaluated indirectly by analysis of the naïve T cell phenotype in the periphery and by measurement of thymus size [11,12]. In a previous study, we showed that bovine thymic function is highly variable, changing with growth stage, gender, and environmental factors such as air temperature and ultraviolet irradiation [13]. ...
The thymus is a primary lymphoid organ where the primary T cell repertoire is generated. Thymus atrophy is induced by various conditions, including infectious diseases, glucocorticoid treatment, and poor breeding management. Cattle with thymus atrophy tend to exhibit weak calf syndrome, a condition in which approximately half of neonates die shortly after birth. Calves with thymus atrophy that survive the first month typically contract chronic inflammatory diseases. In this study, we analyzed the populations of the peripheral blood mononuclear cells and thymocytes in calves with thymus atrophy. In addition, we evaluated polarization of master gene and cytokine mRNA expression in peripheral blood CD4+ cells in the calves. The population of CD4+CD8+ cells in thymus of the calves with thymus atrophy was lower than that of control calves. IL10 mRNA expression in peripheral blood CD4+ cells of calves with thymus atrophy was significantly lower than that of control calves. TBX21 mRNA expression in peripheral CD4+ cells of thymus atrophy calves was tended to be higher than that of the control group. In addition, FOXP3 mRNA expression in peripheral CD4+ cells of the thymus atrophy calves was tended to be lower than that of the control calves. Thymus atrophy calves exhibited chronic inflammatory disease leading, in severe situations, to conditions such as pneumonia with caseous necrosis. These severe inflammatory responses likely are due to decreases in IL10 mRNA expression, impairing control of macrophages, one of the main cell fractions of natural immunity.
... In an attempt to reverse/block this vicious thymic involution cycle, a variety of strategies including cytokines, growth factors, hormonal therapies, in addition to adoptive transfer of precursor T cells or castration have been tested and reviewed in the literature [93][94][95][96]. However, few of these interventions stoodout as the most propitious including: (i) sex-steroid ablation (SSA), (ii) keratinocyte growth factor (KGF), (iii) ghrelin (GRL), (iv) IL-7 as well as, and (v) IL-21. ...
... It is a normal physiological phenomenon that thymus weight and size are associated with age; thymus weight and size begin to increase after birth, reach their maximums in adolescence, and thereafter decrease gradually with age (Steinmann et al. 1985). There are far fewer thymocytes produced in the aged thymus than in the young thymus (Henson et al. 2004). At the same time, thymus atrophy leads to developmental retardation of T cells and less efficient thymopoiesis (Sempowski et al. 2002, Goronzy et al. 2007). ...
Objective:
This study aimed to investigate developmental changes of the thymus and intra- thymic IL-1β, IL-6 and TNF-α expression in weaned Sprague-Dawley rats induced by lipopolysac- charide.
Methods:
Forty healthy weaned rats aged 26 days and weighing 83±4 g were randomly and equally divided into two groups. The lipopolysaccharide group was treated daily with a single injection of lipopolysaccharide for 10 consecutive days, and the saline group was treated with an equal volume of sterilized saline. On the 1st, 4th, 7th and 10th day, histological changes and distribu- tion of IL-1β-, IL-6- and TNF-α-positive cells were detected in the thymus by hematoxylin-eosin and immunohistochemistry staining, respectively. Subsequently, the expression levels of IL-1β, IL-6 and TNF-α were evaluated in the thymus by the ELISA method.
Results:
Thymus weight and index were significantly smaller in lipopolysaccharide-treated rats than in saline-treated rats (p⟨0.05), but no substantial changes were found in the thymus microstructure after lipopolysaccharide induction. Moreover, a large number of IL-1β-, IL-6- and TNF-α-positive cells were observed with brownish-yellow color and mainly distributed in the thy- mus parenchyma, both integrated optical density and average optical density increased signifi- cantly in lipopolysaccharide-treated rats than those in saline-treated rats. Compared with the saline group, most of the thymic homogenates had higher levels of IL-1β, IL-6 and TNF-α in the lipopolysaccharide group on different days.
Conclusion:
These findings indicate that the thymus atrophied after lipopolysaccharide induction in weaned Sprague-Dawley rats, and excessive production of intrathymic IL-1β, IL-6 and TNF-α was probably involved in the atrophic process.
... As a result of thymus regression, thymopoiesis is reduced and thymic function decreases with age (Flores et al., 1999;Linton and Dorshkind, 2004;Yang et al., 2009). Thymic function has been indirectly evaluated by analysis of naïve T-cell phenotype in the periphery and by measurement of thymus size (Hazenberg et al., 2001;Henson et al., 2004). It has been reported that thymic function can be evaluated by measurement of T-cell receptor excision circles (TRECs), which are a by-product of T-cell receptor gene rearrangement in the thymus (Douek et al., 1998;Kong et al., 1998). ...
A signal joint T-cell receptor excision circle (sjTREC) is a circular DNA produced by T-cell receptor α gene rearrangement in the thymus. Measurements of sjTREC values have been used to evaluate thymic function. We recently established a quantitative PCR (QPCR) assay of bovine sjTREC. In the present study, we used this QPCR assay to measure the sjTREC value in bovine peripheral blood mononuclear cells and we then evaluated the relationships between sjTREC values and peripheral blood T-cell number, growth stage, gender, and meteorological season. The sjTREC value was highest at the neonatal stage, and its value subsequently decreased with age. On the other hand, the peripheral T-cell number increased with age. The sjTREC value in calves up to 50-days old was significantly higher for males than for females, suggesting that thymic function might differ by gender. In addition, the sjTREC value and the peripheral T-cell number were significantly higher in calves in the summer season than in calves in the winter season. These data suggest that bovine thymic function is highly variable and varies according to the growth stage, gender, and environmental factors such as air temperature or the UV index.
... Thus, mucosal IL-2 treatment reversed age-impaired mucosal immune responses by enhancing mucosal immunity or abrogating unresponsiveness in aged mice (63). Furthermore, recent studies showed that keratinocyte growth factor or IL-7 treatment prevented thymic atrophy (64,65). IL-15 treatment also restored impaired DC function in mesenteric lymph nodes of aged mice (66). ...
Streptococcus pneumoniae (the pneumococcus) causes a major upper respiratory tract infection often leading to severe illness and death in the elderly. Thus, it is important to induce safe and effective mucosal immunity against this pathogen in order to prevent pnuemocaccal infection. However, this is a very difficult task to elicit protective mucosal IgA antibody responses in older individuals. A combind nasal adjuvant consisting of a plasmid encoding the Flt3 ligand cDNA (pFL) and CpG oligonucleotide (CpG ODN) successfully enhanced S. pneumoniae-specific mucosal immunity in aged mice. In particular, a pneumococcal surface protein A-based nasal vaccine given with pFL and CpG ODN induced complete protection from S. pneumoniae infection. These results show that nasal delivery of a combined DNA adjuvant offers an attractive potential for protection against the pneumococcus in the elderly.
... Traditionally, approaches to improve the immune response in the elderly have focused on thymic rejuvenation [129][130][131]. It has been shown that many approaches can enhance thymopoieses including: administration of hormones and cytokines (growth hormone, prolactin, ghrelin, keratinocyte growth factor, vitamin D, and IL-7), androgen ablation, caloric restriction, bone marrow transplantation, and thymic tissue transplantation [131][132][133][134][135][136][137][138][139][140]. In the future, gene therapy and stem cell transplants may yield even better results. ...
The immune system undergoes a complex and continuous remodeling with aging. Immunosenescence results into both quantitative and qualitative changes of specific cellular subpopulations that have major impact on allorecognition and alloresponse, and consequently on graft rejection and tolerance. Here, we are going to review the immunological changes associated with the aging process relevant for transplantation. Interventions to selectively target changes associated with the senescence process seem promising therapeutic strategies to improve transplantation outcome.
... We have found a reversal of thymic atrophy in the PRG. Hirokawa et al. (1992) and Henson et al. (2004) have stated that under certain circumstances the atrophy of the thymus could be reversed. Our findings are coincident with these studies and contribute to stress the concept that thymus is a dynamic immune organ, and if the etiological agent is eliminated from the body, the morphology of the thymus returns to a normal state thereafter. ...
Solanum glaucophyllum (Sg) [= S. malacoxylon] is a calcinogenic plant inducing "Enzootic Calcinosis" in cattle. The 1,25-dihydroxyvitamin D3, its main toxic principle, regulates bone and calcium metabolism and also exerts immunomodulatory effects. Thymocyte precursors from bone marrow-derived progenitor cells differentiate into mature T-cells. Differentiation of most T lymphocytes is characterized not only by the variable expression of CD4/CD8 receptor molecules and increased surface density of the T cell antigen receptor, but also by changes in the glycosylation pattern of cell surface glycolipids or glycoproteins. Thymocytes exert a feedback influence on thymic non-lymphoid cells. Sg-induced modifications on cattle thymus T-lymphocytes and on non-lymphoid cells were analysed. Heifers were divided into 5 groups (control, intoxicated with Sg during 15, 30 or 60 days, and probably recovered group). Histochemical, immunohistochemical, lectinhistochemical and morphometric techniques were used to characterize different cell populations of the experimental heifers. Sg-poisoned heifers showed a progressive cortical atrophy that was characterized using the peanut agglutinin (PNA) lectin that recognizes immature thymocytes. These animals also increased the amount of non-lymphoid cells per unit area detected with the Picrosirius technique, WGA and DBA lectins, and pancytokeratin and S-100 antibodies. The thymus atrophy found in intoxicated animals resembled that of the physiological aging process. A reversal effect on these changes was observed after suppression of the intoxication. These findings suggest that Sg-intoxication induces either directly, through the 1,25-dihydroxyvitamin D3 itself, or indirectly through the hypercalcemia, the observed alteration of the thymus.
... 10 IL-7 has been shown to be a key cytokine in the early stages of thymocyte development and maintenance of the peripheral naïve T cell pool, with IL-7 expression levels greatly reducing with age. 11 The intrathymic level of IL-7 has a critical effect on the production of T cells 12 and provides the ability to restore the immune system by enhancing thymus output and by rejuvenation of the thymus. [13][14][15] Insulin has also been suggested to serve as a thymus growth factor as shown in several experimental studies in mice. ...
It is unclear, whether pediatric patients with type 1 diabetes (T1DM) show immunological alterations typically found in autoimmune conditions resembling immune dysfunction of the thymus, such as decrease of naïve T cells, lower T cell receptor excision circle (TREC) numbers, telomeric erosion, and diminished interleukin-7 (IL-7) levels. Furthermore, it is unknown, whether long-term therapy with insulin, a thymic growth factor, interferes with these changes. Therefore, the aim of this study was to analyze the quantity of the naïve T cell subset and its TREC content, relative telomere length (RTL) of naïve T cells, and peripheral IL-7 levels in patients with recent-onset T1DM (n = 5), long-standing T1DM (n = 33), and age-matched healthy donors (HD) (n = 37). In long-standing T1DM, TREC numbers/CD8+CD45RA+ T cells were enhanced (p < 0.01) compared to HD and correlated with disease duration (p < 0.02), an independent factor for increased thymic output (p < 0.01), and insulin dosage at blood withdrawal (p < 0.05). IL-7 serum levels were elevated in long-standing T1DM (p < 0.001) and positively correlated with TREC numbers (p < 0.01) and disease duration (p < 0.0001). RTLs in CD8+CD45RA+ T cells were significantly increased compared to HD (p < 0.02). Our data suggest that longterm insulin therapy may serve as a driving factor for thymic function and rejuvenation of the naïve T cell compartment. The ability of the immune system to reconstitute the naïve T cell compartment under well-adjusted insulin therapy may be of major importance for recognition of new antigens, response to vaccinations, and defense of infectious complications.
... Traditionally, approaches that improve the immune response in the elderly have focused on thymic rejuvenation (150)(151)(152)(153)(154)(155)(156). It has been shown that many such treatments (the administration of hormones and cytokines [growth hormone, prolactin, ghrelin, keratinocyte growth factor, vitamin D, IL-7]), androgen ablation, caloric restriction, bone marrow transplant, and thymic tissue transplant) can enhance thymopoiesis (157)(158)(159)(160)(161)(162)(163)(164)(165). In the future, gene therapy and stem cell transplants may yield even better results. ...
The elderly represent the fastest growing segment of the population with end-stage organ disease and the use of aged grafts increased exponentially. Since aging of the immune system, or "immunosenscence" is generally associated with weaker immune responses, one might expect the elderly to be less reactive against transplanted organs than younger patients and therefore to showbetter results in terms of transplant outcome. Paradoxically, however, experimental studies and clinical data of organ transplantation show that old age of either the recipient or the donor is associated with poorer outcomes. On the other hand transplant tolerance is easier to be induced in the neonatal period. One potential reason for this discrepancy may lie in the effects of immunosenescence on the induction of tolerance. While the impact of aging on acute and chronic allograft rejection has been extensively studied, its role on establishing transplant tolerance is not well known. Since tolerance is an active process, and not just the absence of an immune response, the immunologic changes associated with the aging process may interfere with graft survival. In experimental and clinical transplantation, most successful tolerance induction protocols have been tested on young individuals, using grafts from young donors. However, some experiments that have utilized aged animals have demonstrated resistance to tolerance induction. Extrapolation of these results to humans suggests that protocols for clinical tolerance induction may not be effective in the elderly and may need to be revised for this population. The resistance to achieving immunological tolerance with aging is complex and multifactorial. Here, we review the age associated changes that may interfere with immunologic tolerance. Understanding this phenomenon may help in developing novel therapeutic approaches to reverse the crucial dysfunctions of the aging immune system and achieve effective tolerance regimens for the elderly.
... IL-7 has been shown to be a key cytokine in the early stages of thymocyte development and maintenance of the peripheral T cell pool, 120 with IL-7 expression levels greatly reducing with age. 121 This reduction is associated with a decrease in thymic size, cellularity, and output. IL-7 was suggested to be one of the components responsible, among others, for higher probability of reaching extreme ages. ...
The thymus is the main source of recent thymic emigrants (RTE) and naïve T cells. The aging of the immune system (immunosenescence) is characterized by loss of thymic function, decreased numbers of RTE, peripheral proliferation of mature T cells, and oligoclonal expansions of specific T cell subpopulations. As shown in several studies, thymectomized patients demonstrate signs of premature immunosenescence reminiscent of aged people, such as decreased proportions of naïve T cells and RTE, a compensatory increase of mature T cell subpopulations with increased proliferation rates, restriction of the T cell receptor repertoire, and a delayed response to new antigens and vaccinations. This review demonstrates that, despite some limitations, childhood thymectomy may serve as an useful model for premature immunosenescence, mimicking changes expected after physiological thymus involution in the elderly. Thus, it may prove an insightful tool for obtaining better understanding of human naïve T cell development, thymic function, and maintenance of the naïve T cell pool.
... T cells Function Declines Ernst et al. (1990), Miller (1996), Garcia and Miller (1997), Pawelec et al. (1999) Aukrust et al. (1996), Kim and Nel (2005) Naïve CD4 + T cells IL-2 production Reduced Meydani et al. (1990), Engwerda et al. (1996), Linton et al. (1996Linton et al. ( , 2005, Kurashima and Utsuyama (1997) Regulatory T cells Increased Sharma et al. (2006) Montecino- Rodriguez et al., 1998;Aspinall and Andrew, 2000;Andrew and Aspinall, 2001) but also for potential therapeutic approaches in human patients (Napolitano et al., 2002;Henson et al., 2004;Sutherland et al., 2005;Herndler-Brandstetter et al., 2008). Homeostatic proliferation, induced either by aging-dependent or disease-associated thymic involution and lymphopenia, has, vice versa, been made responsible for a higher incidence of autoimmunity in humans (Mishra and Kammer, 1998;Douek et al., 2003;Piliero et al., 2004;Jameson, 2005;Thewissen et al., 2005;Giovannetti et al., 2008), as it results in the clonal expansion of memory (-like) T cells that reduce the diversity of the available TCR repertoire (Berzins et al., 2002;Kovaiou et al., 2005;Herndler-Brandstetter et al., 2008). ...
Deregulation of reactive oxygen intermediates (ROI) resulting in either too high or too low concentrations are commonly recognized to be at least in part responsible for many changes associated with aging. This article reviews ROI-dependent mechanisms critically contributing to the decline of immune function during physiologic - or premature - aging. While ROI serve important effector functions in cellular metabolism, signalling and host defence, their fine-tuned generation declines over time, and ROI-mediated damage to several cellular components and/or signalling deviations become increasingly prevalent. Although distinct ROI-associated pathomechanisms contribute to immunosenescence of the innate and adaptive immune system, mutual amplification of dysfunctions may often result in hyporesponsiveness and immunodeficiency, or in chronic inflammation with hyperresponsiveness/deregulation, or both. In this context, we point out how imbalanced ROI contribute ambiguously to driving immunosenescence, chronic inflammation and autoimmunity. Although ROI may offer a distinct potential for therapeutic targeting along with the charming opportunity to rescue from deleterious processes of aging and chronic inflammatory diseases, such modifications, owing to the complexity of metabolic interactions, may carry a marked risk of unforeseen side effects.
... As an immunotherapeutic agent, IL-7 can augment T cell numbers by 1) increasing thymic output (Henson et al., 2004), 2) promoting T-cell survival (Kim et al., 1998;Khaled and Durum, 2003), and 3) inducing cell cycling (Geiselhart et al., 2001;Kittipatarin et al., 2006). While the activity of IL-7 as a survival factor promoting thymic development has been well documented (Maraskovsky et al., 1997;Khaled et al., 1999), its function as a proliferative factor remains poorly understood. ...
Interleukin-7 (IL-7) increases lymphocyte numbers, a critical feature of immune reconstitution, through mechanisms that are still poorly understood. Part of the problem is that IL-7 is produced in limited amounts by non-lymphoid cells, making in vivo studies of the cytokine's activity a challenge. To overcome this, we developed an in vitro system by which lymphocytes from secondary immune organs could be cultured to produce IL-7 responsive cells. Using this method, we showed that CD8(hi)CD44(hi) T cells accumulate in culture with IL-7 from a population of lymph node or splenic cells. These results were validated when a similar lymphocyte subset was found in mice expressing a constitutively active form of STAT5b, a key transducer of IL-7 signals. Interestingly, IL-7-expanded cells also up regulated the activation marker, CD69. The IL-7-derived CD44(hi)CD69(hi) cells were not generated from naïve cells, but expanded from an existing population, since culture in IL-7 of naïve lymphocytes from OT-1/Rag1(-/-) mice did not produce CD44(hi)CD69(hi) cells. Using the in vitro culture system to study lymphocytes from mice deficient in the apoptotic protein, BIM, we were able to attribute the expansion of CD8(hi)CD44(hi)CD69(hi) T cells to the proliferative and not survival activity of IL-7. The in vitro culture system provides an important new methodology to examine the activities of this essential as well as immunotherapeutic cytokine.
... A ging is associated with a decline in most physiological functions, among them the immunological system. This process is often named immunosenescence and is related, for example, to an impaired capacity to generate new T cells (10). It has been postulated that intense and prolonged exercise may be associated with a postexercise window of opportunity for infection because of a decrease in the activity of natural killer cells as well as blood and salivary levels of immunoglobulin (21,23,30). ...
It has been suggested that elderly people are more susceptible to infectious diseases because of immunosenescence. The well-described transient immunosuppressive effect of exercise may increase health risks in such populations. Although resistance training has been recommended to older individuals, little is known regarding its acute effects on immune indexes. This study examined the acute effects of resistance exercise sessions performed at different intensities (50 vs. 80% of one-repetition maximum [1RM]) on salivary immunoglobulin A (IgA), total leukocytes, lymphocytes and their subsets, and the cortisol levels of older women. Fifteen physically independent elderly women (mean age = 67.5 +/- 3.9 years) underwent, in a randomized manner, two experimental sessions comprising resistance exercise for the major muscular groups or one control session without exercise. Session 1 consisted of two sets of 13 repetitions at 50% 1RM, whereas session 2 consisted of two sets of eight repetitions at 80% 1RM. Blood and saliva samples were collected at rest, immediately after, and at 3 and 48 hours after the completion of the sessions. Blood cortisol, total leukocytes, lymphocytes and their subpopulations, and salivary IgA were measured by standard techniques. Overall, resistance exercise sessions failed to provoke significant decline on the examined immune function indexes. Instead, resistance exercise sessions induced significant elevation in salivary IgA levels, but this was not observed after the control session. Also, experimental sessions did not induce significant elevations in blood cortisol. It was concluded that resistance exercise sessions performed at 50 or 80% of 1RM do not present suppressive effects on the studied immune parameters. Conversely, a transient elevation was observed in salivary IgA levels, which requires support in future investigations. Taken together, the observed results suggest that elderly women can engage in resistance training programs without negative acute effects on the studied parameters of the immune system.
... over time but cannot multiply, they remain within T lymphocytes as they transit and are diluted during T cell proliferation (23, 24). The reduction in thymopoiesis witnessed in aging is paralleled by a decrease in thymic output (15,25) as measured by increased dilution of TRECs. ...
Nutritional status is highly dependent on season in countries such as The Gambia. In a rural Gambian setting, individuals born during periods of seasonal nutritional deprivation ("hungry seasons") are susceptible to mortality from infectious diseases in adult life.
We investigated the hypothesis that impaired immunocompetence in those born in the hungry season results from an underlying defect in immunologic memory, similar to the immunosenescence of old age, which is likely to be reflected in the phenotype and kinetics of T lymphocytes in young adults.
T cell phenotype in terms of CD3, CD4, CD8, CD45RA, and CD45R0 expression and in vivo dynamics measured by stable isotope labeling of T cell subsets combined with gas chromatography-mass spectrometry and frequency of T cell receptor excision circles were measured in 25 young (18-24-y-old) Gambian men. Thirteen of these 25 men were exposed to perinatal malnutrition as defined by birth season and birth weight.
In persons born in the hungry season with low birth weight, no differences in the proportions of memory or naive T cells were found. Kinetic analysis showed higher proliferation rates in memory (CD45R0(+)) subsets of T cells than in naïve (CD45R0(-)) cells, which is consistent with previous studies, but no evidence was found for an effect of birth weight or season on T lymphocyte proliferation and disappearance rates. No significant correlations were found between in vivo T cell kinetics and frequency of T cell receptor excision circles. Only absolute numbers of granulocytes were elevated in those born in the nutritionally deprived season.
In healthy young Gambian men, T lymphocyte homeostasis is extremely robust regardless of perinatal nutritional compromise.
Bei juveniler idiopathischer Arthritis konnte eine prämature Immunoseneszenz nachgewiesen werden. Ein Erklärungsmodell ist, dass die prämature Immunoseneszenz den primären Defekt darstellt, der das Immunsystem zum Versagen der Selbsttoleranz führt. Eine andere Deutungsmöglichkeit stellt die prämature Immunoseneszenz als Folge von chronischer Stimulation und Aktivierung des Immunsystems durch die Autoimmunerkrankung selbst dar. In dieser Arbeit wurden die Immunoseneszenz-Parameter (naive T-Zellen, RTE, IL-7-Level, TRECs, relative Telomerlänge, Ki-67-Expression) von JIA-Patienten - unterteilt in eine DMARD-Gruppe und eine TNFα-Inhibitor-Gruppe - mit denen von gesunden Vergleichsprobanden verglichen. Eine fortgeschrittenere Immunoseneszenz bei den Gesunden könnte möglicherweise durch vermehrte chronische Virusinfekte erklärt werden, die jedoch in dieser Arbeit nicht erfasst wurden. In der vorliegenden Arbeit konnte eine potenzielle Rekonstitutionsfähigkeit mit Verbesserung der Immunoseneszenz-Parameter bei DMARD-Therapie demonstriert werden. So trat ein höherer Anteil an naiven T-Zellen und an RTE auf, was vermuten lässt, dass der fortschreitende Verlust der Thymusfunktion bei Patienten mit Autoimmunerkrankung reversibel sein könnte. Bei Vergleich der TNFi-Patienten mit der DMARD-Gruppe konnte eine weiter fortgeschrittene Immunoseneszenz festgestellt werden. Dies könnte durch die Ansammlung von schwereren, DMARD-refraktären Patienten, aber auch durch die unterschiedliche, aber nicht-signifikante Altersverteilung bedingt sein. Bei längerer Einnahmedauer des TNFα-Inhibitors zeigte sich ein tendenziell stärkeres Auftreten von naiven T-Zellen und RTE, kombiniert mit geringeren Anteilen differenzierterer Subpopulationen. So scheinen TNFα-Inhibitoren die Fähigkeit zu besitzen, die prämature Immunoseneszenz positiv zu beeinflussen oder zumindest eine Verlangsamung des prämaturen Alterungsvorgangs zu bewirken.
In this chapter we describe changes in the immune system that are thought to be related to age per se. We subsequently review the clinical implications of these changes, including the effects of surgical trauma on immune function (see the physiology table at beginning of chapter). We then discuss how stress modifies many of these changes. We also describe recent information on persistent infections, in particular latent viral infections and how they may be partly responsible for shaping the aging immune system. We conclude with a discussion of some of the latest research on ways to restore or stimulate immune function in the elderly.
Human aging is associated with sharp increases in the frequency of degenerative diseases, autoimmune diseases, and cancer, and a multitude of physiological age changes occur in parallel. The differentiation between physiological aging and age-related diseases is exceedingly difficult, however, and the pathogenic mechanisms are incompletely understood, which complicates the characterization of the age-related changes which truly contribute to disease pathogenesis. The aging immune system is characterized by features which resemble autoimmune diseases such as rheumatoid arthritis. The possible contribution of immunosenescence to autoimmunity and the mechanisms of both degeneration and possibly regeneration of the immune system are discussed.
Nutritional status is highly dependent on season in countries such as The Gambia. In a rural Gambian setting, individuals born during periods of seasonal nutritional deprivation ("hungry seasons") are susceptible to mortality from infectious diseases in adult life. OBJECTIVE: We investigated the hypothesis that impaired immunocompetence in those born in the hungry season results from an underlying defect in immunologic memory, similar to the immunosenescence of old age, which is likely to be reflected in the phenotype and kinetics of T lymphocytes in young adults. DESIGN: T cell phenotype in terms of CD3, CD4, CD8, CD45RA, and CD45R0 expression and in vivo dynamics measured by stable isotope labeling of T cell subsets combined with gas chromatography-mass spectrometry and frequency of T cell receptor excision circles were measured in 25 young (18-24-y-old) Gambian men. Thirteen of these 25 men were exposed to perinatal malnutrition as defined by birth season and birth weight. RESULTS: In persons born in the hungry season with low birth weight, no differences in the proportions of memory or naive T cells were found. Kinetic analysis showed higher proliferation rates in memory (CD45R0(+)) subsets of T cells than in naïve (CD45R0(-)) cells, which is consistent with previous studies, but no evidence was found for an effect of birth weight or season on T lymphocyte proliferation and disappearance rates. No significant correlations were found between in vivo T cell kinetics and frequency of T cell receptor excision circles. Only absolute numbers of granulocytes were elevated in those born in the nutritionally deprived season. CONCLUSION: In healthy young Gambian men, T lymphocyte homeostasis is extremely robust regardless of perinatal nutritional compromise.
The thymus is a central lymphoid organ critical for the development and maintenance of an effective peripheral T-cell repertoire. Most important, it provides a specialized environment for the selection of rearranged clones that will function appropriately in the adaptive immune response. Thymic involution has been observed in several model systems; including graft-versus-host disease, aging, viral infection, and tumor development, however, the precise mechanisms involved in this phenomenon remain poorly defined. Here, we review some of our results related to the studies of the cell-mediated immunity in a mammary tumor model; more specifically, those related to the tumor-induced impaired T-cell development and thymic involution. Collectively, the understanding of the mechanisms and pathways associated with the tumor-induced thymic involution is essential for the development of innovative and safe therapies to fight against the immune suppression caused by the tumor development.
This chapter summarizes recent work suggesting that human immunosenescence may be closely related to both psychological distress
and stress hormones. The age-related immunological changes are also similarly found during chronic stress or glucocorticoid
exposure. It follows that endogenous glucocorticoids (cortisol) could be associated to immunosenescence. When compared with
young subjects, healthy elders are emotionally distressed in parallel to increased cortisol/ dehydroepiandrosterone (DHEA)
ratio. Furthermore, chronic stressed elderly subjects may be particularly at risk of stress-related pathology because of further
alterations in glucocorticoid-immune signaling. Although DHEA and its metabolites have been described with immune-enhancing
properties, their potential use as hormonal boosters of immunity should be interpreted with caution. The psychoneuroendocrine
hypothesis of immunosenescence is presented in which the agerelated increase in the cortisol/DHEA ratio is major determinant
of immunological changes observed during aging. We finally discuss that strictly healthy elders are largely protected from
chronic stress exposure and show normal cortisol levels and T-lymphocyte function. This information adds a new key dimension
on the biology of aging and stress.
With the improvement of medical care and hygienic conditions, there has been a tremendous increment in human lifespan. However, many of the elderly (>65 years) display chronic illnesses, and a majority requires frequent and longer hospitalization. The robustness of the immune system to eliminate or control infections is often eroded with advancing age. Nevertheless, some elderly individuals do cope better than others. The origin of these inter-individual differences may come from genetic, lifestyle conditions (nutrition, socio-economic parameters), as well as the type, number and recurrence of pathogens encountered during life. The theory we are supporting is that chronic infections, through life, will induce profound changes in the immune system probably due to unbalanced inflammatory profiles. Persistent viruses such a cytomegalovirus are not eliminated and are a driven force to immune exhaustion. Because of their age, elderly individuals may have seen more of these chronic stimulators and have experienced more reactivation episodes ultimately leading to shrinkage of their repertoire and overall immune robustness. This review integrates updates on immunity with advancing age and its impact on associated clinical conditions.
Apoptosis is a programmed cell death that represents a normal component of the development, differentiation and health of multicellular organisms leading to an adequate cellular turnover and homeostasis. In autoimmune diseases, the immune system recognizes various autoantigens causing damage in target organs. Dead cells represent an important source of autoantigens that, in particular conditions, can represent a stimulus for an autoimmune response. A large number of studies reported the impairment of the apoptosis regulatory mechanisms in immune cells as a pivotal element in the pathogenesis and evolution of autoimmune disorders. Several pathogenetic pathways have been claimed to account for autoimmunity development during apoptotic processes. In fact, interestingly abnormalities potentially leading to immune disorders have been described as occurring in each step involved in apoptosis, from the very beginning to the post death phenomena. In this extent we propose a systematic review of the molecular mechanisms strictly leading to apoptosis with particular interest to their alterations, potentially causing tissue specific and/or systemic autoimmunity.
The ubiquitin-proteasome pathway, which degrades intracellular proteins, is involved in numerous cellular processes, including the supply of immunocompetent peptides to the antigen presenting machinery. Proteolysis by proteasomes is conducted by three beta subunits, beta1, beta2, and beta5, of the 20S proteasome. Recently, a novel beta subunit expressed exclusively in cortical thymic epithelial cells was discovered in mice. This subunit, designated beta5t, is a component of the thymoproteasome, a specialized type of proteasomes implicated in thymic positive selection. In this study, we show that, like its mouse counterpart, human beta5t is expressed exclusively in the thymic cortex. Human beta5t was expressed in approximately 80% of cortical thymic epithelial cells and some cortical dendritic cells. Human beta5t was incorporated into proteasomes with two other catalytically active beta subunits beta1i and beta2i, forming 20S proteasomes with subunit compositions characteristic of thymoproteasomes. The present study demonstrates, for the first time, the existence of thymoproteasomes in the human thymic cortex, indicating that thymoproteasome function is likely conserved between humans and mice.
This manuscript reviews current evidence suggesting that aging of the immune system (immunosenescence) may be closely related to chronic stress and stress factors. Healthy aging has been associated with emotional distress in parallel to increased cortisol to dehydroepiandrosterone (DHEA) ratio. The impaired DHEA secretion together with the increase of cortisol results in an enhanced exposure of lymphoid cells to deleterious glucocorticoid actions. The lack of appropriated growth hormone signaling during immunosenescence is also discussed. It follows that altered neuroendocrine functions could be underlying several immunosenescence features. Indeed, changes in both innate and adaptive immune responses during aging are also similarly reported during chronic glucocorticoid exposure. In addition, chronically stressed elderly subjects may be particularly at risk of stress-related pathology because of further alterations in both neuroendocrine and immune systems. The accelerated senescent features induced by chronic stress include higher oxidative stress, reduced telomere length, chronic glucocorticoid exposure, thymic involution, changes in cellular trafficking, reduced cell-mediated immunity, steroid resistance, and chronic low-grade inflammation. These senescent features are related to increased morbidity and mortality among chronically stressed elderly people. Overall, these data suggest that chronic stress leads to premature aging of key allostatic systems involved in the adaptation of the organisms to environmental changes. Stress management and psychosocial support may thus promote a better quality of life for elderly people and at the same time reduce hospitalization costs.
This paper reviews recent work suggesting that human immunosenescence may be closely related to both chronic stress and stress hormones. The age-related immunological changes are also similarly found during chronic stress or glucocorticoid exposure. These data further suggest that endogenous glucocorticoids could be associated with immunosenescence. When compared with young subjects, healthy elders are emotionally distressed in parallel to increased cortisol/dehydroepiandrosterone ratio. Furthermore, chronically stressed elderly subjects may be particularly at risk of stress-related pathology because of further alterations in glucocorticoid-immune signaling. Age-related increase in cortisol/dehydroepiandrosterone ratio could be understood as a major determinant of immunological changes observed during aging. Strictly healthy elders are somewhat protected from chronic stress exposure and show normal cortisol levels and increased T cell function. This information adds a new key dimension to the biology of aging and stress.
Evidence is accumulating that elderly individuals are more susceptible to infection with organisms to which they were previously immune. This indicates that there might be a limit to the persistence of immune memory. This fact is particularly disturbing because the average life expectancy of humans has almost doubled in the past 200 years and is still increasing. We discuss mechanisms that might constrain the persistence of memory T cells and consider whether humans will suffer from memory T-cell exhaustion as life expectancy increases.
Currently, the aging research field lacks consensus in its focus and methodology. Foundational principles, such as the evolutionary origins and physiological definition of aging, remain controversial. The aim of this paper is to resolve these issues. By applying the concepts of thermodynamics and information in an evolutionary context, the aging phenotype can be derived from first principles. Life uses information storage to maintain its distance from thermodynamic equilibrium. Since it is impossible to make any process 100% efficient, a selective force (i.e., natural selection) is needed to maintain the information's viability. Natural selection operates upon generations, and for reasons discussed subsequently, the somatic body cannot implement an analogous selective process. The aging phenotype we see can be derived from this model along with a number of insights that will enhance our ability to make intelligent and rational interventions.
Ageing has been associated with immunological changes (immunosenescence) that resemble those observed following chronic stress or glucocorticoid (GC) treatment. These changes include thymic involution, lower number of naïve T cells, reduced cell-mediated immunity, and poor vaccination response to new antigens. It follows that immunosenescence could be associated with changes of peripheral GC levels. Indeed, when compared with young subjects, healthy elders are more stressed and show activation of the hypothalamus-pituitary-adrenal (HPA) axis. However, both beneficial and undesirable effects of GCs ultimately depend on the target tissue sensitivity to these steroids. Recent data indicate that peripheral lymphocytes from elders respond poorly to GC treatment in vitro. The present review summarizes recent findings which suggest that immunosenescence may be closely related to both psychological distress and stress hormones. Furthermore, chronically stressed elderly subjects may be particularly at risk of stress-related pathology because of further alterations in GC-immune signalling. Finally, the neuroendocrine hypothesis of immunosenescence is finally reconsidered in which the age-related increase in the cortisol/DHEA ratio is major determinant of immunological changes observed during ageing.
Thymic function is important for the generation of T-cell diversity in the periphery of both children and adults during both health and disease. Until recently, thymic function could not be monitored, as a consequence of the absence of adequate technology to differentiate recent thymic emigrants (RTEs) from naïve T cells. The generation of TCR diversity occurs in the thymus through recombination of gene segments encoding the variable parts of the TCR alpha and beta chains. During these processes, by-products of the rearrangements are generated in the form of signal joint T-cell receptor excision circles (sjTRECs). As sjTRECs are stable extrachromosomal DNA fragments, they are not replicated during mitosis and thus diluted with each round of cell division, and are therefore most frequent in naïve T cells that have recently left the thymus, their quantification is actually considered as a valuable tool to estimate thymic function. Therefore, quantitative sjTRECs content have been recently used to assess thymic output during both health and disease. In this review, we summarize recent data on the recent thymic output function feature in patients with hematological malignancy and the immune reconstitution after stem cell transplantation and we also characterize factors that may improve the thymic output function.
The T cell system is a complex and highly dynamic system that is amazingly robust over many decades of human life. Its functional competence is determined not only by its size but also by its diversity. Homeostatic control mechanisms have to secure sufficient T cell replenishment while preventing loss of clonal diversity. Major homeostatic challenges include profound expansion and shrinkage of T cell clonotypes upon antigenic triggering and, more importantly, age-related changes in T cell regeneration. The ability of the thymus to rebuild a diverse repertoire ceases in the fifth decade of life. Emerging data suggest that the end of the 7th decade of life defines a critical time period when T cell homeostasis is no longer guaranteed and diversity of the naïve T-cell repertoire collapses. Thus, failure of T cell homeostasis appears to result from cumulative defects of T cell generation. Elucidation of the underlying mechanisms will allow for extending this turning point to later in life; ultimately, interventions have to aim at restoring thymic function and complementary modes of T cell reconstitution.
Aging of the immune system, or immunosenescence, is characterized by changes in T cell subsets, cellular and molecular level alterations and thymic atrophy, resulting in a decline of T and B cell function. These alterations have been shown to be accompanied by a loss of ability to recognize "self" and "foreign" antigens. Therefore the development of autoimmune responses like production of autoantibodies has been hypothesized to be secondary to thymus involution with a decline of naïve T cells and accumulation of clonal T cells with activation due to "neoantigens" during the aging process. Altered apoptosis and altered T cell homeostasis have been emphasized to promote a chronic inflammatory state and lead to the concept of a immune-risk phenotype. However, it has to be proven which kinds of mechanisms turn the immune system to manifest autoimmune disease and how speculated defects in T cell differentiation and interaction leading to premature aging of the immune system may contribute to the development of autoimmune diseases.
In vivo administration of alcoholic extract of medicinal plant Tinospora cordifolia (TC) to mice bearing a spontaneous T cell lymphoma designated as Dalton's lymphoma prevented tumor growth-dependent regression of thymus. TC was found to augment proliferation of thymocytes with a concomitant decrease in thymocyte apoptosis. It also resulted in a decrease in the number of Hassal's corpuscles. Restoration of thymus homeostasis was caused by TC-dependent augmentation in production of thymocyte growth promoting cytokines Interleukin-2 and Interferon-gamma from thymocytes. TC was found to downregulate thymocyte apoptosis by modulation of Caspase pathway. TC administration retarded tumor growth and prolonged survival of tumor-bearing mice. The possible mechanisms are discussed.
Decline in immune function with advancing age is a hallmark of aging. As a result, infectious diseases cause more morbidity and mortality to the elderly. These age-related changes significantly alter the functionality of both innate and adaptive immune components.
Although age-associated dysfunction of the immune system is well reported, not much is known about the extrinsic and intrinsic factors that progressively bring about functional changes in immune cell types.
This review focuses on our current understanding of age-associated changes in innate and adaptive immune cell types and how this knowledge could be used to improve the immune competence of elderly individuals.
Therapeutic approaches to rejuvenate the exhausted immune system in the elderly need more attention.
The thymus represents the primary site for T cell lymphopoiesis, providing a coordinated set for critical factors to induce and support lineage commitment, differentiation and survival of thymus-seeding cells. One irrefutable fact is that the presence of non-lymphoid cells through the thymic parenchyma serves to provide coordinated migration and differentiation of T lymphocytes. Moreover, the link between foetal development and normal anatomy has been stressed in this review. Regarding thymic embryology, its epithelium is derived from the embryonic endodermal layer, with possible contributions from the ectoderm. A series of differentiating steps is essential, each of which must be completed in order to provide the optimum environment for thymic development and function. The second part of this article is focused on thymic T-cell development and differentiation, which is a stepwise process, mediated by a variety of stromal cells in different regions of the organ. It depends strongly on the thymic microenvironment, a cellular network formed by epithelial cells, macrophages, dendritic cells and fibroblasts, that provide the combination of cellular interactions, cytokines and chemokines to induce thymocyte precursors for the generation of functional T cells. The mediators of this process are not well defined but it has been demonstrated that some interactions are under neuroendocrine control. Moreover, some studies pointed out that reciprocal signals from developing T cells also are essential for establishment and maintenance of the thymic microenvironment. Finally, we have also highlighted the heterogeneity of the lymphoid, non-lymphoid components and the multi-phasic steps of thymic differentiation.
T cells can be divided into two groups on the basis of the expression of either alpha beta or gamma delta T-cell receptors (TCRs). Because the TCR delta chain locus lies within the larger TCR alpha chain locus, control of the utilization of these two receptors is important in T-cell development, specifically for determination of T-cell type: rearrangement of the alpha locus results in deletion of the delta coding segments and commitment to the alpha beta lineage. In the developing thymus, a relative site-specific recombination occurs by which the TCR delta chain gene segments are deleted. This deletion removes all D delta, J delta, and C delta genes and occurs on both alleles. This delta deletional mechanism is evolutionarily conserved between mice and humans. Transgenic mice which contain the human delta deleting elements and as much internal TCR delta chain coding sequence as possible without allowing the formation of a complete delta chain gene were developed. Several transgenic lines showing recombinations between deleting elements within the transgene were developed. These lines demonstrate that utilization of the delta deleting elements occurs in alpha beta T cells of the spleen and thymus. These recombinations are rare in the gamma delta population, indicating that the machinery for utilization of delta deleting elements is functional in alpha beta T cells but absent in gamma delta T cells. Furthermore, a discrete population of early thymocytes containing delta deleting element recombinations but not V alpha-to-J alpha rearrangements has been identified. These data are consistent with a model in which delta deletion contributes to the implementation of a signal by which the TCR alpha chain locus is rearranged and expressed and thus becomes an alpha beta T cell.
It has been generally believed that human CD8+ memory cells are principally found within the CD45ROhigh population. There are high frequencies of CD8+ memory CTL specific for the human CMV tegument phosphoprotein pp65 in PBMC of long-term virus carriers; the large population of memory CTL specific for a given pp65 peptide contains individual CTL clones that have greatly expanded. In this study, we found high frequencies of pp65 peptide-specific memory CTL precursors in the CD45ROhighCD45RA- population, but also appreciable frequencies in the CD45RAhigh subpopulation. Because the majority of CD8+ T cells in PBMC are CD45RAhigh, more of the total pp65-specific memory CTL pool is within the CD45RAhigh than in the CD45ROhigh compartment. Using clonotypic oligonucleotide probes to quantify the size of individual pp65-specific CTL clones in vivo, we found the CD45RAhigh population contributed 6- to 10-fold more than the CD45ROhigh population to the total virus-specific clone size in CD8+ cells. During primary CMV infection, an individual virus-specific CTL clone was initially CD45ROhigh, but after resolution of infection this clone was detected in both the CD45ROhigh and the CD45RAhigh populations. We conclude that CD45RA+ human CD8+ T cells do not solely comprise naive cells, but contain a very significant proportion of memory cells, which can revert from the CD45ROhigh to CD45RAhigh phenotype in vivo.
In this essay we suggest that the primary goal of the cells of the immune system is to ensure their own growth and survival. In adults, in steady-state conditions, the number and distribution of lymphocyte populations is under homeostatic control. New lymphocytes that are continuously produced in primary and secondary lymphoid organs must compete with resident cells for survival. We discuss recent findings supporting lymphocyte survival as a continuous active process and implicating cognate receptor engagement as fundamental survival signals for both T and B lymphocytes. The conflict of survival interests between different cell types gives rise to a pattern of interactions that mimics the behavior of complex ecological systems. In their flight for survival and in response to competition, lymphocytes use different survival signals within different ecological niches during cell differentiation. This is the case for T and B lymphocytes and also for naive and memory/activated T and B cells. We discuss how niche differentiation allows the co-existence of different cell types and guarantees both repertoire diversity and efficient immune responses.
Recent thymic emigrants can be identified by T cell receptor excision circles (TRECs) formed during T-cell receptor rearrangement. Decreasing numbers of TRECs have been observed with aging and in human immunodeficiency virus (HIV)-1 infected individuals, suggesting thymic impairment. Here, we show that in healthy individuals, declining thymic output will affect the TREC content only when accompanied by naive T-cell division. The rapid decline in TRECs observed during HIV-1 infection and the increase following HAART are better explained not by thymic impairment, but by changes in peripheral T-cell division rates. Our data indicate that TREC content in healthy individuals is only indirectly related to thymic output, and in HIV-1 infection is mainly affected by immune activation.
Thymic atrophy is an age-associated decline in commitment to the T cell lineage considered to be associated with defective TCR beta-chain rearrangement. Both IL-7 and stem cell factor (SCF) have dominant roles at this stage of triple negative (TN) thymocyte development. Because there is no age-associated decrease in the number of CD44(+)CD25(-)CD3(-)CD4(-)CD8(-) cells, this study investigated whether alterations in apoptosis within the TN pathway accounted for diminishing thymocyte numbers with age. Here we show significant age-associated increases in apoptotic TN thymocytes, specifically within CD44(+)CD25(+) and CD44(-)CD25(+) subpopulations, known to be the location of TCR beta-chain rearrangement. IL-7 added to TN cultures established from old mice significantly both reduces apoptosis and increases the percentage of live cells within CD44(+)CD25(+) and CD44(-)CD25(+) subpopulations after 24 h, with prosurvival effects remaining after 5 days. SCF failed to demonstrate prosurvival effects in old or young cultures, and IL-7 and SCF together did not improve upon IL-7 alone. IL-7R expression did not decline with age, ruling out the possibility that the age-associated increase in apoptosis was attributed to reduced IL-7R expression. Compared with PBS, treatment of old mice with IL-7 produced significant increases in live TN cells. By comparison, treatment with SCF failed to increase live TN numbers, and IL-7 and SCF together failed to significantly improve thymopoiesis above that shown by IL-7 alone. Thus, treatment with IL-7 alone can reverse the age-associated defect in TN thymocyte development revealed by in vitro studies to be located at the stages of TCR beta-chain rearrangement.
Interleukin-7 (IL-7) is important for thymopoiesis in mice and humans because IL-7 receptor alpha (IL-7Ralpha) mutations result in a severe combined immunodeficiency phenotype with severe thymic hypoplasia. Recent evidence has indicated that IL-7 also plays an important role as a regulator of T-cell homeostasis. Here we report the immunologic effects of recombinant human IL-7 (rhIL-7) therapy in normal and simian immunodeficiency virus (SIV)-infected nonhuman primates. Cynomolgus monkeys receiving 10 days of rhIL-7 showed substantial, reversible increases in T-cell numbers involving a dramatic expansion of both naive and nonnaive phenotype CD4(+) and CD8(+) subsets. Although IL-7 is known to have thymopoietic effects in mice, we observed marked declines in the frequency and absolute number of T-cell receptor excision circle-positive (TREC(+)) cells in the peripheral blood and dramatic increases in the percentage of cycling T cells in the peripheral blood as measured by Ki-67 expression (baseline less than 5% to approximately 50% after 6 days of therapy) and ex vivo bromodeoxyuridine (BrdU) incorporation. Similarly, moderately CD4- depleted SIV-infected macaques treated with rhIL-7 also had significant increases in peripheral blood CD4(+) and CD8(+) T cells following rhIL-7 therapy. Thus, rhIL-7 induces dramatic alterations in peripheral T-cell homeostasis in both T-cell-replete and T-cell-depleted nonhuman primates. These results further implicate IL-7 as a promising immunorestorative agent but illustrate that a major component of its immunorestorative capacity reflects effects on mature cells. These results also raise the possibility that IL-7 therapy could be used to temporarily modulate T-cell cycling in vivo in the context of immunotherapies such as vaccination.
TCR-beta gene rearrangement or expression is necessary and sufficient for the progression of early alpha beta thymocyte differentiation from the CD3-CD4-CD8- triple negative (TN)3 to the CD4+CD8+ double positive stage. The onset of TCR-beta rearrangement is currently thought to occur gradually. Some thymocytes were reported to be rearranged at the earliest (CD44+CD25-) TN stage, whereas other thymocytes did not initiate TCR-beta rearrangement until the latest (CD44-CD25-) TN stage. Here, we have isolated subsets of TN thymocytes on the basis of surface expression of CD44 and CD25, with c-kit as an additional marker. We present a revised model of early T cell development in which TCR-beta and TCR-gamma rearrangements occur abruptly, at the CD44lowCD25+ c-kitlowTN stage. A high level of c-kit expression defines pro-T cells which have not yet rearranged their TCR genes. Germ-line TCR-beta transcripts, and transcripts of recombination activating genes (RAG)-1 and 2, are detected before TCR-beta gene rearrangement. Analyses of TN thymocytes of RAG-1 mutant mice, and of various TCR mutant and TCR transgenic RAG-1 mutant mice, indicate the existence of a control point at the CD44-CD25+TN stage at which cells expressing a productively rearranged TCR-beta chain are selected for further differentiation.
Despite playing a critical role in the development of naive T cells, the thymus is involuted with age. Whether a single age-associated defect or multiple aberrations contribute to thymic involution remains controversial. Here, we determined molecular aberrations in the thymocyte and epithelium compartments of the aging thymus. We demonstrated that total thymocyte numbers declined with a stepwise kinetics; clear demarcations occurred at 1.5, 3, 12 and 22 months of age. By quantitative PCR, a 2.4-fold reduction in the copies of signal joint TCR-excised circle (sjTREC)/10 5 thymocytes was first detected at 3 months; no further reduction observed thereafter. Nevertheless, the combined reductions in thymocyte numbers and sjTREC/10 5 cells caused a 7-fold decrease in sjTREC/thymus by 3 months, 21-fold by 18 months and 72-fold by 22 months as compared to 1 month. We showed aberration in expression of E2A, a transcription regulator critical for TCRb rearrangement. While E2A expression declined 3-fold by 3 months and 18-fold by 7 months, expression of LMO2, a negative regulator of E2A activities, increased 5-fold by 18 months. Interestingly, expression of pre-Ta and its transcriptional regulator HEB were not reduced with age. Furthermore, keratin-8 expression, specific for cortical thymic epithelium, declined 3-fold by 7 months and remained stable thereafter. In contrast, Foxn1 expression was reduced 3-fold by 3 months, 16-fold by 12 months and 37-fold by 18 months. IL-7 expression was not reduced until 7 months and reached 15-fold reduction by 22 months. Thus, the data demonstrate that thymic involution results not from a single defect, but culminates from an array of molecular aberrations in both the developing thymocytes and thymic epithelials.
The thymus is the primary organ responsible for the production of mature TCR α / β T cells. Quantification of a DNA excision circle that is produced during TCR rearrangement, termed a signal joint TCR rearrangement excision circle (sjTREC) can be used as a measure of thymic function. Here sjTREC measurement has been applied to two monkey species used as animal models of human disease, rhesus macaques (Asian origin) and sooty mangabeys (African origin). Initial PCR analysis determined that the TCR δRec-ΨJα rearrangement leading to sjTREC formation occurs in both species. Primers to a DNA sequence conserved in macaques, mangabeys and humans were used in a quantitative competitive PCR assay to quantify sjTREC. We found that as in humans, sjTREC in these two monkey species decline with age. sjTREC are first generated in thymocytes during the early stages of TCR rearrangement. Lymph node CD4+ and CD8+ T cells contain more sjTREC than peripheral blood T cell populations, suggesting that recent thymic emigrants home to the lymphoid tissues. The sjTREC level is significantly higher within the peripheral blood CD4+ and CD8+ T cells of mangabeys compared to macaques. Removal of the thymus in four macaques led to a profound decrease in peripheral blood sjTREC level by 1 year post-thymectomy, indicating the lack of a significant extra-thymic source of peripheral naive T cells in macaques. Our results indicate that production, trafficking, and proliferation of recent thymic emigrants in these two monkey species represents a useful animal model system for understanding human immunological disorders.
The aim of this study was to investigate the specific role of endogenous glucocorticoids (GC) following restraint stress on thymic involution and apoptosis. Restraint stress has been reported to alter physiological and behavioral responses in experimental animals. Exposure of mice to restraint stress led to involution of the thymus, to a decrease of the CD4+ 8+ thymocyte subset, and to fragmentation of thymic DNA. The role of endogenous GC in restraint stress-induced changes in the thymus was studied by three experimental approaches: surgical adrenalectomy, chemical adrenalectomy, and blocking of GC receptors by a specific type II receptor antagonist. In surgically-Adx mice, which lack endogenous GC, the effects of restraint on the thymus were wholly abrogated. Pretreatment of restrained mice with metyrapone (an 11beta hydroxylase inhibitor that specifically inhibits GC biosynthesis) had the same consequence, and blockage of GC receptors with the specific GC type II receptor antagonist RU-38486 attenuated the effects of the stressor. These findings indicate that GC are involved in the restraint-induced effects on the thymus.
A continuous but low input of stem cells or 'prothymocytes' is necessary to maintain T-cell development in the adult thymus, but the colonizing cell has not been characterized. Precursors of T cells have been found in the minor CD4-8- population of thymocytes, but even the earliest cells of this population already have partially rearranged T-cell antigen receptor (TCR) genes. We now demonstrate that the thymus contains a minute population of lymphoid cells similar in some but not all respects to bone marrow-derived haemopoietic stem cells. This population has TCR genes in a germline state. It gives a slow but extensive reconstitution of both alpha beta and gamma delta lineages on transfer into an irradiated thymus, with kinetics indicating that it includes the earliest intrathymic precursor cells so far isolated. Surprisingly, these cells express low surface levels of the mature T-cell marker CD4.
T cells express T-cell antigen receptors (TCR) for the recognition of antigen in conjunction with the products of the major histocompatibility complex. They also express two key surface coreceptors, CD4 and CD8, which are involved in the interaction with their ligands. As CD4 is expressed on the early haemopoietic progenitor as well as the early thymic precursor cells, a role for CD4 in haemopoiesis and T-cell development is implicated. Thymocytes undergo a series of differentiation and selection steps to become mature CD4+8- or CD4-8+ (single positive) T cells. Studies of the role of CD4+ T cells in vivo have been based on adoptive transfer of selected or depleted lymphocytes, or in vivo treatment of thymectomized mice with monoclonal antibodies causing depletion of CD4+ T cells. In order to study the role of the CD4 molecule in the development and function of lymphocytes, we have disrupted the CD4 gene in embryonic stem cells by homologous recombination. Germ-line transmission of the mutation produces mutant mouse strains that do not express CD4 on the cell surface. In these mice, the development of CD8+ T cells and myeloid components is unaltered, indicating that expression of CD4 on progenitor cells and CD4+ CD8+ (double positive) thymocytes is not obligatory. Here we report that these mice have markedly decreased helper cell activity for antibody responses, although cytotoxic T-cell activity against viruses is in the normal range. This differential requirement for CD4+ helper T cells is important to our understanding of immune disorders including AIDS, in which CD4+ cells are reduced or absent.
The terminal involution pattern of the human thymus was studied based on autopsy cases (both sexes, age range 63-91 years). Large sections through the entire thymic fat body were examined with the help of both conventional histological and immunohistochemical techniques. The findings demonstrate that thymic atrophy in old humans (a) goes far beyond the degree of involution observed in small rodents; (b) results in a system of thin, branching, in part interrupted, non-keratinizing epithelial plates containing no typical Hassall bodies; (c) concerns all components of the thymus except fat tissue which progressively replaces original thymic structures; and (d) involves various types of disorganization of individual lobules with T and B lymphocytes often located outside rather than within epithelial remnants. Effects of low-level radiation on this final regression of the human thymus are unknown.
Patients undergoing cardiac bypass operations normally have a thymectomy to facilitate cannulation of the great vessels. Laboratory indices of immune function were measured in 18 children aged 9 months to 3 years who had had a thymectomy when aged 3 months or less, and in two groups of controls individually matched for age and age at operation. Total lymphocyte numbers were similar in all three groups but thymectomized children had significantly lower numbers of T cells and T cell sub-sets than controls and showed diminished responses to phytohaemagglutinin and concanavalin A. Children who have had a thymectomy early in life represent an important group in the study of the development of the immune system in man. Although the clinical consequences of early thymectomy are unclear, evidence of impairment of parameters of immunity have been found in later infancy and routine thymectomy in paediatric cardiac surgery should be avoided.
Although the biological mechanisms of aging are poorly understood, many clinical observations have suggested a close relation between senescence and immunity. Apart from a progressive decrease in muscular power and sensory perception, and loss of cells from most organs and tissues, aging is paralleled by an increasing vulnerability to infections and an increasing liability to malignant tumors and autoimmune conditions. Late-life high-incidence diseases, including vascular disease, maturity-onset diabetes, cancer, amyloidosis, and senile dementia, all exhibit malfunctions of the immune system or are somehow related to the immune system (WALFORD 1980). Speculations and theories about aging and immunity are manifold. Their rationales range from explaining aging by a committed progressive breakdown of functions of immunocompetent cells and their clonal exhaustion (WALFORD 1969; BURNET 1970; HOLLIDAY et al. 1981) to explaining progressive loss of immunity by phenomena of aging such as: (a) somatic mutation (ORGEL 1963, 1973); (b) error in replication or repair of DNA (JOHNSON and STREHLER 1972; HART and SETLOW 1974); (c) error proneness of DNA polymerases and related enzymes (BURNET 1976); or (d) respiration-dependent injury to the mitochondrial genome (FLEMING et al. 1982) in other cells than immunocompetent cells. Several excellent recent reviews have compiled mutual associations between age and immune system of different species, including man (WALFORD 1969; MAKINODAN et al. 1971; KAY and MAKINODAN 1976; MAKINODAN and YUNIS 1977; YUNIS et al. 1978; GOOD et al. 1979; KISHIMOTO and MITSUYA 1980; WILLIAMS et al. 1980; Leech 1980; WEKSLER 1980; MAKINODAN 1980; KAY and MAKINODAN 1981). In particular, T cell system-dependent immune functions seem to decline with age (Table 1), whereas functions of macrophages and B cells do not appear to be impaired (YUNIS et al. 1978; KAY and MAKINODAN 1981; BENNER et al. 1981).
One hundred and thirty-six thymuses completely removed at autopsy from persons suffering a sudden death were examined by stereological and morphometrical methods. Adding biopsy material from immunologically healthy cardiac patients we obtained relative volumes from 204 persons ranging in age from 1 month to 107 years. The size of the human thymus remains unchanged during ageing under normal physiological conditions (median: 19.5 cm3). Individual maximum size (range: 5-70 cm3) is reached in the first year of life. Early histological changes are in the enlargement of the perivascular space, the Hassall's bodies, and the connective tissue. This begins in the first year of life, reaches a maximum from 10 to 25 years, then declines again. Adipose tissue replaces the lymphocytic perivascular space and the connective tissue only. This occurs extensively after the age of 15 years. When defined by the silver impregnation technique, the volumes of the thymic epithelium (cortex and medulla), show a continuous involution from the first year to the end of life. The curve can be approximated to simple negative logarithmic functions. The velocity and nature of involution of the thymic epithelium do not change under the influence of the changing hormonal balance due to puberty. Since important thymic functions (T lymphopoiesis and T-cell differentiation) are located in the epithelial space, the age-related involution of the human thymus is not related to puberty.
TCR-beta gene rearrangement or expression is necessary and sufficient for the progression of early alpha beta thymocyte differentiation from the CD3-CD4-CD8- triple negative (TN)3 to the CD4+CD8+ double positive stage. The onset of TCR-beta rearrangement is currently thought to occur gradually. Some thymocytes were reported to be rearranged at the earliest (CD44+CD25-) TN stage, whereas other thymocytes did not initiate TCR-beta rearrangement until the latest (CD44-CD25-) TN stage. Here, we have isolated subsets of TN thymocytes on the basis of surface expression of CD44 and CD25, with c-kit as an additional marker. We present a revised model of early T cell development in which TCR-beta and TCR-gamma rearrangements occur abruptly, at the CD44lowCD25+ c-kitlowTN stage. A high level of c-kit expression defines pro-T cells which have not yet rearranged their TCR genes. Germ-line TCR-beta transcripts, and transcripts of recombination activating genes (RAG)-1 and 2, are detected before TCR-beta gene rearrangement. Analyses of TN thymocytes of RAG-1 mutant mice, and of various TCR mutant and TCR transgenic RAG-1 mutant mice, indicate the existence of a control point at the CD44-CD25+TN stage at which cells expressing a productively rearranged TCR-beta chain are selected for further differentiation.
The diversity of the T cell receptor repertoire is generated by rearrangement of gene elements in immature thymocytes. To
identify a thymic signal that induces this rearrangement, a variety of agents were tested for their ability to induce rearrangement
of the T cell receptor beta gene in suspensions of thymocytes from mouse embryos at day 14 of gestation. Of 16 agents tested,
only interleukin-7 (IL-7) induced V(D)J gene rearrangement and sustained expression of the RAG-1 and RAG-2 genes, which are
known to control rearrangement. These data implicate IL-7, a cytokine that is abundantly expressed in embryonic thymus, in
driving gene rearrangement during early T cell development.
Studies of the functional properties and developmental potentials of immediate post-thymic cells have been hampered by the lack of reliable markers with which to distinguish recent thymic emigrants (RTE) from the bulk of peripheral T cells. In the present study, the intrathymic FITC-labeling technique was used in concert with three-color flow-cytometric analysis to identify, phenotypically characterize, and study the short term fate of RTE in normal rats. The results indicated that between 3 and 4% of total T cells in lymph node and spleen of 5- to 8-wk-old rats had been released from the thymus within the preceding 24 h. Unlike the bulk of the peripheral T cells, which had a Thy1-, CD45RC+, and/or RT6+ phenotype, these RTE were Thy1+, CD45RC-, and RT6-. Furthermore, two discrete subsets of RTE were defined: a major subset (approximately 95%) of CD4+ or 8+ (single positive), TCR-alpha beta hi T cells that resembled medullary thymocytes; and a minor subset (approximately 5%) of CD4+ and 8+ (double positive), TCR-alpha beta low T cells that resembled cortical thymocytes. The following evidence suggested that most if not all Thy1+ T cells in young adult rats are RTE and their immediate descendants: 1) thymectomy (but not sham thymectomy) selectively depleted Thy1+ T cells from lymph node within 3 to 7 days, even in adrenalectomized rats; 2) most FITC-labeled RTE differentiated into Thy1-, CD45RC+, RT6+ T cells within 7 days of release from the thymus; 3) transitional phenotypes of Thy1+ T cells, including Thy1low, CD45RC+, and RT6+, were observed in normal, as well as in intrathymic, FITC-injected rats; 4) most T cells in neonatal rats were Thy1+ and RT6-, whereas their descendants were Thy1- and RT6+. These experiments demonstrate that most RTE in normal rats are phenotypically (and presumably developmentally) immature at the time of release from the thymus, and progressively acquire the phenotypic attributes of more mature T cells post-thymically. These unique phenotypic attributes should expedite the isolation of RTE and their immediate descendants for definitive studies of their developmental and functional properties.
We have subdivided mouse CD4-CD8-CD3- triple-negative (TN) thymocytes into four subsets based upon expression of CD44 and CD25, including CD44+CD25-, CD44+CD25+, CD44-CD25+ and CD44-CD25-. Characterization of these cells revealed several features distinct to each subset, in particular the expression of high levels of c-kit (the receptor for stem cell factor) by CD44+CD25-TN and CD44+CD25+TN but not by CD44-CD25+TN and CD44-CD25-TN. The CD44+CD25+TN subset also included the IL-7 and stem cell factor-responsive cells, whereas only minimal responsiveness was observed by the CD44- populations. These subsets also showed differential cytokine production potential (CD44+CD25- > CD44+CD25+ > CD44-CD25+ > CD44-CD25-) after stimulation with calcium ionophore, PMA and IL-1. The repopulation potential of these subsets in 2-deoxyguanosine-treated fetal thymic lobes supports the following maturation sequence: CD44+CD25- -->CD44+CD25+ -->CD44-CD25+ -->CD44-CD25-. Furthermore, the sequence of progression from CD44+CD25+ to CD44-CD25+ cells was confirmed by their TCR beta-chain gene configuration. The former population exhibits germ-line TCR beta-chain configuration, whereas the latter subset shows a rearranged pattern.
While early thymic T cell precursor populations and their maturational sequence have been recently identified, the signals driving differentiation are unknown. While cytokines may play an integral role in T cell development, various mouse models rendered genetically deficient for specific cytokines do not display abnormalities in T cell development. Recently, we have generated IL-7 -/- mice and reported that IL-7 plays a unique and nonredundant role in lymphopoiesis. These mice displayed a 10- to 20-fold reduction in the total number of T and B cells. Here, we show that IL-7 -/- mice display a sharp reduction in both the frequency and absolute number of adult thymic gamma delta T cells while retaining normal frequencies of alpha beta T cells. This defect in gamma delta T cell production extends to peripheral organs as IL-7 -/- mice are essentially devoid of splenic and intestinal intraepithelial gamma delta T cells. This aberrant phenotype was traced back to impaired fetal gamma delta T cell maturation. In the absence of IL-7, differentiation of immature V gamma 3 low-CD24+ fetal T cells to mature V gamma 3 high CD24- cells is inhibited. In contrast, NK cell maturation appears to be only mildly affected in the absence of IL-7. To further clarify the role of IL-7 in thymic development, detailed analysis of CD3-4-8- thymic precursors was performed. A partial inhibition in the differentiation of CD44+25+ pro-T cells into CD44-25+ pre-T cells was observed. Unexpectedly, the lack of IL-7 resulted in decreased expression of CD117 (c-kit) on both CD4 low and pro-T cells, suggesting that IL-7 may influence the expression of other cytokine receptors involved in early hemopoietic development. Together, these data clarify the developmental abnormalities during T cell development due to the absence of IL-7.
The embryonic thymic microenvironment provides the necessary elements for T cell lineage commitment, but the precise role of individual stromal cell components remains to be determined. Here we address the question of which stromal cell types are required for initiation of V-DJ rearrangements of the TCR-beta and TCR-delta locus in CD117+CD45+ uncommitted fetal liver progenitors. We show that fetal thymic stroma alone is necessary and sufficient for induction of TCR-beta and TCR-delta rearrangements. Furthermore, the ability to induce this T cell commitment step is confined to a subset of MHC class II-positive epithelial cells. Thymic stroma derived from mice with a targeted deletion in the IL-7 gene, however, lacks this ability. These findings set the stage for a further definition of the nature of the thymic stromal cell support in the regulation of T cell commitment.
Growth factors have been implicated in thymocyte development, but mutants lacking cytokines, or their receptors, have failed to reveal essential roles for growth/differentiation factors in the thymus. Mutations in the receptor tyrosine kinase c-kit and the common cytokine receptor gamma chain (gamma c) reduce cellularity, but are permissive for thymocyte development. We now report that thymocyte development is completely abrogated in mice lacking both c-kit and gamma c (c-kit-gamma c-). Thymic hypocellularity is so severe that the T cell receptor repertoire fails to form except for monoclonal or oligoclonal beta chain DJ rearrangements. B lymphopoiesis is only mildly reduced in c-kit-gamma c- as compared with c-kit+gamma c- mice, and hematological values are identical comparing c-kit-deficient and c-kit-gamma c- mice. These experiments reveal essential, overlapping, and synergistic functions for two distinct signaling pathways, one utilizing c-kit and the other cytokine receptor gamma c complexes coupling to Janus kinases and signal transducers and activators of transcription.
Involution of the thymus is a feature of age and precedes inefficient functioning of the immune system. C57BL/10 mice show an 83% reduction in number of thymocytes between 3 and 20 mo of age, with a significant decline in each of the thymic subsets defined by their expression of CD4 and CD8. The similar percentage contribution of each subset to the whole at 3, 12, and 20 mo suggests a lesion in the T cell developmental pathway within an early subset. The CD3- CD4- CD8- subset showed a significant decline in number by 20 mo of age, but despite this reduction, no significant difference was noted in the number of CD44+ CD25- cells, the earliest stage of this subset between 3 and 20 mo of age. A significant decline in the number of their progeny, the CD44+ CD25+, and the progeny of these cells, the CD44- CD25+ cells, was noted by 12 mo of age. Expression of CD25 within this subset is associated with rearrangement of TCR beta-chain genes. F5 transgenic mice, carrying a complete TCR-alphabeta transgene under the control of a CD2 minigene cassette on a C57BL/10 background, showed no age-associated thymic atrophy in any of the defined thymic subsets over the same period as the normal C57BL/10 mice. Similar results were noted with mice carrying the same transgene but which in addition were also RAG-1-. The results indicate that age-associated thymic involution was associated with problems with rearrangement of the TCR beta-chain genes affecting the production of thymocytes.
The pre-T cell receptor (pre-TCR) that minimally consists of the TCR beta chain and the disulfide-linked pre-T cell receptor alpha (pT alpha) chain in association with signal-transducing CD3 molecules rescues from programmed cell death cells with productive TCR beta rearrangements. The pre-TCR induces expansion and differentiation of these cells such that they become TCR alpha beta bearing CD4+8+ thymocytes, which express only a single TCR beta chain and then either die of neglect or--upon TCR-ligand interaction--undergo either positive or negative selection. The newly discovered pT alpha gene encodes a transmembrane protein that belongs to the Ig superfamily and contains a cytoplasmic tail that, however, has no essential function in signal transduction, which is mediated by CD3 molecules and most likely p56lck. Experiments in pT alpha gene-deficient mice show that the pre-TCR has a crucial role in maturation as well as allelic exclusion of alpha beta T cells but is not required for the development of gamma delta-expressing cells. The function of the pre-TCR cannot be fully assumed by an alpha beta TCR that is expressed abnormally early in T cell development.
Expression of the avian chT1 thymocyte antigen persists on a subpopulation of peripheral T cells enriched in the DNA deletion circles created by alphabeta and gammadelta TCR gene rearrangements. The chT1+ cells are evenly distributed among all of the peripheral T lymphocyte compartments. The levels of chT1+ T cells in the periphery gradually decline in parallel with age-related thymic involution, and these cells disappear following early thymectomy. Experiments in which variable numbers of the 14 thymic lobes are removed in young chicks indicate a direct correlation between the levels of circulating chT1+ cells and residual thymic mass. Measurement of recent thymic emigrants in the periphery thus provides an accurate indication of thymic function.
Age-associated thymic involution results in a diminished capacity to regenerate T cell populations, although the magnitude of this effect is unknown. In this report, thymic function was studied in aged vs. young adult mice after lethal irradiation and administration of T cell-depleted bone marrow (BM) from young mice. Abnormalities observed in aged thymi (reduced thymocyte numbers, histologic abnormalities) were not reversed by administration of young BM via bone marrow transplantation (BMT), but aged thymi displayed a normal thymocyte subset distribution and appropriately deleted MIs-reactive T cells after BMT. Aged BMT recipients regenerated significantly reduced numbers of splenic T cells compared to young recipients and showed increased peripheral expansion of thymic emigrants since a higher proportion of BM-derived T cells expressed a memory phenotype in aged vs. young BMT recipients. Because peripheral expansion of thymic emigrants could substantially increase the number of thymic progeny present in the spleen, we sought to measure thymic T cell regenerative capacity after BMT in a setting devoid of peripheral expansion. To do this, TCR-transgenic (Tg+) T cell-depleted BM was administered to aged and young recipients lacking antigen specific for the Tg+ TCR. Aged recipients regenerated approximately 50 % of the TCR Tg+ cells regenerated in young BMT recipients, providing evidence that even very aged thymi retain the capacity to regenerate significant numbers of mature T cell progeny. Therefore, thymic function is reduced with aged but it is not lost, suggesting that therapeutic approaches to enhance thymic function may be successful even in very aged hosts.
The thymus represents the major site of the production and generation of T cells expressing alphabeta-type T-cell antigen receptors. Age-related involution may affect the ability of the thymus to reconstitute T cells expressing CD4 cell-surface antigens that are lost during HIV infection; this effect has been seen after chemotherapy and bone-marrow transplantation. Adult HIV-infected patients treated with highly active antiretroviral therapy (HAART) show a progressive increase in their number of naive CD4-positive T cells. These cells could arise through expansion of existing naive T cells in the periphery or through thymic production of new naive T cells. Here we quantify thymic output by measuring the excisional DNA products of TCR-gene rearrangement. We find that, although thymic function declines with age, substantial output is maintained into late adulthood. HIV infection leads to a decrease in thymic function that can be measured in the peripheral blood and lymphoid tissues. In adults treated with HAART, there is a rapid and sustained increase in thymic output in most subjects. These results indicate that the adult thymus can contribute to immune reconstitution following HAART.
To determine whether the thymus is still functional despite age-related involution, we measured a biomarker for thymopoiesis known as the T cell receptor excision circle (TREC) from peripheral blood mononuclear cells (PBMCs) of 148 healthy children and from PBMCs, CD4(+), and CD8(+) cells of 32, 30, and 50 healthy adults, respectively. We demonstrate that during the first 5 years of life, thymic output is decreased (P 0.002) but not dramatically (r = -0. 282). Among adults aged 23-58, thymic output was inversely correlated with age, as measured from PBMCs (r = -0.628, P < 0.0005), CD4(+) (r = -0.530, P 0.003), and CD8(+) fractions (r = -0.385, P 0. 006). A strong correlation existed between pediatric PBMC TRECs and the expression of three naïve phenotypic markers (CD45RA(+)CD45RO(-), CD45RA(+)CD62L(+), and CD45RO(-)CD27(+)CD95(low)). Adult PBMC TRECs correlated only with the expression of CD45RA(+)CD45RO(-) (r = 0.459, P 0.012). Our data suggest that in adults CD45RA(+)CD45RO(-) may be enriched for TRECs and add to a growing body of evidence illustrating intact thymic function in adulthood.
Age-associated thymic atrophy results in a decline in T lymphocyte output and has been identified as one of the key events that precede inefficient functioning of the immune system in later life. Thymic atrophy is thought to result from a failure of the thymic microenvironment to support thymopoiesis in old age and recent evidence suggests that a decline in interleukin-7 (IL-7) expression may limit thymocyte development by restricting combinations of survival, proliferation and rearrangement of the TCRbeta chain. Using RT-PCR and the RNase protection assay, we show that the expression of IL-7 declines with age. Analysis of Connexin 43 expression, a component molecule of gap junctions, whose function is to connect epithelial cells, does not markedly decline with age. These observations suggest that a decline in IL-7 expression is not matched by a similar loss of epithelial cells. These results in conjunction with other studies lead us to speculate that IL-7 producing MHC class II positive TECs are being replaced by cells that do not have this capacity.
Signal joint T cell receptor delta (TCRD) excision circles (TRECs) are episomal DNA circles generated by the DNA recombination process that is used by T lymphocytes to produce antigen-specific alpha/beta T cell receptors. Measurement of TRECs in thymocytes and peripheral blood T cells has been used to study thymus output in chickens and humans. We have developed a real-time quantitative-PCR assay for the specific detection and quantification of mouse TCRD episomal DNA circles excised from the TCRA locus during TCRA gene rearrangement (mTRECs). We found that the mouse TCRD TRECs detected with this assay were predominantly in naïve phenotype CD4(+) and CD8(+) T cells. In a series of aged mice (range 6-90-week-old) we determined the absolute number of thymocytes and the number of molecules of mTRECs/100,000 thymocytes. We found that the absolute number of thymocytes dramatically decreased with age (P<0.05) and that molecules of mTREC/100,000 thymocytes also declined with mouse age (P<0.05). Splenocytes were isolated from aging mice and the frequency of naïve phenotype CD4 and CD8 cells determined. There was a significant drop in both CD4 and CD8 naïve peripheral T cells in the aged mice over time. mTREC analysis in purified CD4(+) and CD8(+) splenocytes demonstrated a constant level of mTRECs in the CD4 compartment until age 90 weeks, while the mTRECs in the CD8 compartment fell with age (P<0.05). By combining the mouse TREC assay with T cell phenotypic analysis, we demonstrated that IL-7 administration to young mice induced both increased thymopoiesis and peripheral T cell proliferation. In contrast, IL-7 treatment of aged mice did not augment thymopoiesis, nor induce expansion of splenic T cells. Thus, thymus output continues throughout murine adult life, and the thymic atrophy of aging in mice is not reversed by administration of IL-7.
The human thymus exports newly generated T cells to the periphery. As no markers have been identi???ed for these recent thymic
emigrants (RTE), it is presently impossible to measure human thymic output. T cell receptor excision circles (TREC) have been
recently used to assess thymic output during both health and disease. Using a mathematical model, we quantify age-dependent changes both in the number of RTE generated per day and in TREC concentration during an 80-year lifespan. Through analyses, we demonstrate that RTE and peripheral T cell division have the same potential to affect TREC concentration at any age in healthy people. T cell death also in???uences TREC concentration, but to a lesser extent. During aging, our results indicate that thymic involution primarily induces an age-dependent decline in TREC concentrations within both CD4+ and CD8+ T cell populations. We further apply this model for studying TREC concentration during HIV-1 infection. Our analyses reveal that a decrease in thymic output is the major contributor to the decline in TREC concentration within CD4+ T cells, whereas both increased peripheral T cell division and decreased thymic output induce the decline in TREC concentration within CD8+ T cells. Therefore, we suggest that T cell turnover should be examined together with TREC concentration as a measure of RTE. If peripheral T cell division remains relatively unchanged, then TREC concentration indeed re???ects thymic output.
Despite playing a critical role in the development of naive T cells, the thymus is involuted with age. Whether a single age-associated defect or multiple aberrations contribute to thymic involution remains controversial. Here, we determined molecular aberrations in the thymocyte and epithelium compartments of the aging thymus. We demonstrated that total thymocyte numbers declined with a stepwise kinetics; clear demarcations occurred at 1.5, 3, 12 and 22 months of age. By quantitative PCR, a 2.4-fold reduction in the copies of signal joint TCR-excised circle (sjTREC)/10(5) thymocytes was first detected at 3 months; no further reduction observed thereafter. Nevertheless, the combined reductions in thymocyte numbers and sjTREC/10(5) cells caused a 7-fold decrease in sjTREC/thymus by 3 months, 21-fold by 18 months and 72-fold by 22 months as compared to 1 month. We showed aberration in expression of E2A, a transcription regulator critical for TCR beta rearrangement. While E2A expression declined 3-fold by 3 months and 18-fold by 7 months, expression of LMO2, a negative regulator of E2A activities, increased 5-fold by 18 months. Interestingly, expression of pre-T alpha and its transcriptional regulator HEB were not reduced with age. Furthermore, keratin-8 expression, specific for cortical thymic epithelium, declined 3-fold by 7 months and remained stable thereafter. In contrast, Foxn1 expression was reduced 3-fold by 3 months, 16-fold by 12 months and 37-fold by 18 months. IL-7 expression was not reduced until 7 months and reached 15-fold reduction by 22 months. Thus, the data demonstrate that thymic involution results not from a single defect, but culminates from an array of molecular aberrations in both the developing thymocytes and thymic epithelials.
Aging of the murine thymus is accompanied by a measurable loss of weight and cellularity and a marked reduction in its output of T lymphocytes. This study employs a molecular approach to determine changes in the output of the murine thymus using a novel assay system based on the detection and quantitation of the excised TCRdelta DNA locus from within the TCRalpha chain genes. In alpha beta(+) T cells such delta excision circles (deltaEC) are present at higher levels in naive T cells as compared with memory T cell populations, are non-replicating, and diluted within the total peripheral alpha beta(+) T cell pool with advancing age. This assay permits the assessment of thymic output in older animals where previous analysis was hampered by the transient nature of the naive T cell surface phenotype, and so allows the assessment of the efficacy of IL-7 as an agent to reverse thymic atrophy. Treatment of old mice with IL-7 although producing no overall change in the total number of alpha beta(+) T cells in the peripheral T cell pool altered the component subsets. Mice treated with IL-7 showed increases in the number of alpha beta(+)TCR cells possessing deltaEC commensurate with improved thymic output, and the splenic T cells from IL-7-treated mice performed significantly better in in vitro functional assays compared to those from age-matched saline-treated controls.