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PFP expression in PBL. Human PBL were stained with anti-PFP Ab. PFP immunoreactivity is observed in the cytoplasm of human PBL. x 800.
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Perforin (PFP) is a cytotoxic protein released from killer cells. PFP immunoreactivity in human peripheral blood lymphocytes (PBL) and tumour infiltrating lymphocytes (TIL) was investigated immunocytochemically with the aid of an anti-PFP monoclonal antibody. PFP was detected in the cytoplasm of 10% of PBL. We performed a double staining of PFP+ ce...
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... PFP + cells in PBL Specific PFP immunostaining was observed in the cytoplasm of lymphocytes (Figure 1). PFP was detected in large lym- phocytes and accounted for about 10% of PBL. Preimmune rat serum or preabsorbed antibody used as a control did not yield specific staining. These control tests were repeated on the cytocentrifuged specimens of PBL with the same results. Double staining showed that 18% of CD16 + cells and 9% of CD8 + cells but none of the CD4 + cells produced PFP (Figure ...
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... cancer (PC), perforin expression in CTLs and NK cells was diminished 11 . In colorectal cancer, the number of perforin expressing CD8+ or CD16+ cells was lowered, which correlated with tumor progression 12 . For these reasons, perforin could be an attractive therapeutic target in cancer immunotherapy. ...
Perforin secreted from cytotoxic lymphocytes plays a critical role in cancer immunosurveillance. The aim of this study was to investigate the therapeutic potential of liposomes containing perforin expression vector driven by the promotor of prostate-specific antigen (PSA). The anti-tumor effect of perforin was analyzed using prostate cancer (PC) PC-3 cells in which perforin expression was controlled by Tet-on system (PC-3PRF cells). Liposomes encapsulating PSA promoter-driven perforin expression vector (pLipo) were constructed for its specific expression in PC. The anti-tumor effect of pLipo was evaluated in vitro using docetaxel-resistant PC 22Rv1 PC cell line, 22Rv1DR, and PC-3 cells in the presence of human peripheral blood mono nuclear cells (PBMCs) and also in vivo using male nude mice bearing 22Rv1DR cell-derived tumor xenograft. Induction of perforin significantly inhibited growth of PC-3PRF cells. Treatment with pLipo induced perforin expression in 22Rv1DR cells expressing PSA but not in PC-3 cells lacking it. Treatment with pLipo at a low concentration was prone to inhibit growth of both cell lines and significantly inhibited growth of 22Rv1DR cells when co-incubated with PBMCs. The combined use of pLipo at a high concentration with PBMCs showed nearly complete inhibition of 22Rv1DR cell growth. Intravenous administration of pLipo via tail vein increased the level of perforin in tumor and serum and significantly decreased the tumor volume. Our results suggest that liposome-mediated PC-specific expression of perforin could be a novel therapy for advanced PC.
... double coloring of pfp+cells has shown that pfp in Til (tumor-infiltrating lymphocyte) mainly produces cd8+ cells but not cd16 cells which were a different medical finding from that from pbl (peripheral blood lymphocyte). nakanishi et al. explain that pfp+ cells were most numerous in dukes A and decreased in number according to the progression of tumours (12). The pfp+ cells in Til exhibited the same phenotypes as those in pbl but the pfp+ cells were more numerous in cd8+ cells than in cd16+ cells at all stages. ...
... This study represents the first evidence that pfp is mainly secreted from cd8+ cells in tumour tissues. it is hypothesised that the decrease in the number of pfp+ cells in accordance with tumour progression may reflect the suppression of the hosts local immunity (12). ...
Some literature reviews have been carried out about the role of perforin in medicine. The first step involved a systematic search to identify relevant studies published between 2001 and 2019 in the following electronic databases - EBSCO host, Scopus, Science Direct, Web of Science, and Elsevier. By analyzing the available literature, it can be concluded that perforin plays an important role in cytoxical activity of natural killer cells (NK) and CD8+ T cell. NK and CD8+use the same mechanism for destroying target cells. This article cites the disease hemophagocytic lymphohistiocytosis (HLH) which is characterized by heavy abnormalities in the immune system. The point is that this disease is caused by perforin gene mutation. The key is the application of properly sensitized dendritic cells (DCs) because they are effective in immunotherapy against cancer. It may be effective in γ-irradiated colon cancer cell lines HT-29. Growth hormone inhibiting hormone (GIH) induces maturation and activation of DCs. In that way, GIH-Dcs shows increased cytotoxic activity and higher perforin and granzyme expression. So, this means that theoretical research has shown that efficient activity against cancer is induced when DCs are sensitized with γ-irradiated cancer cells. In that way, through a direct increase of cytotoxicity and indirect T cell activation,there can beanti-tumor activity. It is suggested to continue scientific research about the role of perforin in the future.
... Its deregulated expression has been analyzed in many cancer related studies. For example, in colorectal cancer tissues, the decrease in the number of perforin positive cells in accordance with tumor progression may reflect the suppression of the hosts local immunity (Nakanishi, Monden, Morimoto, et al., 1991). Another study revealed that co-expression of GrmB and perforin leads to the focus formation inhibition and apoptosis activation in Hep-2 cells and provided evidence for potential therapeutic applications in laryngeal cancer . ...
Apoptosis, also named programmed cell death, is a fundament process required for morphogenetic homeostasis during early development and in pathophysiological conditions. It is come into existence in 1972 by work of Kerr, Wyllie and Currie and later on investigated during the research on development of the C. elegans. Trigger by several stimuli, apoptosis is necessary during the embryonic development and aging as homeostatic mechanism to control the cell population and also play a key role as defense mechanism against the immune responses and elimination of damaged cells. Cancer, a genetic disease, is a growing burden on the health and economy of both developing and developed countries. Every year there is tremendously increasing in the number of new cancer cases and mortality rate. Although, there is a significant improvement have been made in biotechnological and bioinformatic fields however, the therapeutic advantages and cancer etiology is still under explored. Several studies determined the deregulation of different apoptotic components during the cancer development and progression. Apoptosis relies on activation of distinct signaling pathways that are often deregulated in cancer. Thus, exploring the single or more than one apoptotic component underlying their expression in carcinogenesis could help to track the disease progression. Current book chapter will provide the several evidences supporting the use of different apoptotic components as prognosis and prediction markers in various human cancer types.
... 10 Infiltrating T cells also included cytotoxic T cells (CTL) carrying cytoplasmic perforin granules. 11 Our observations suggest an occurrence of host anti-tumor immune responses in colorectal cancer that affects prognosis. ...
T cell infiltration in colorectal cancer is associated with a favorable prognosis, suggesting an occurrence of a certain degree of anti-tumor immunity. T helper type 1 (Th1) and Th2 cells are now known to selectively express CC-chemokine receptor 5 (CCR5)/CXC-chemokine receptor 3 (CXCR3) and CCR4, respectively. To clarify the mechanism of T cell infiltration, we examined in situ expression of these chemokine receptors and their respective chemokine ligands in 40 cases of human colorectal cancer. Immunohistochemistry showed a predominant accumulation of T cells expressing CCR5 and CXCR3 mainly along the invasive margin, whereas those expressing CCR4 were rare. Flow cytometric analysis showed that more than half of CD8(+) T cells and a fraction of CD4(+) cells isolated from fresh tumor tissues co-expressed CCR5 and CXCR3, and CD8(+) T cells and CD4(+) cells predominantly produced interferon-gamma (IFN-gamma) over interleukin-4 (IL-4) after in vitro stimulation. RANTES/CCL5, a ligand of CCR5, was localized within infiltrating CD8(+) T cells in a granular pattern, whereas IP-10/CXCL10, a ligand of CXCR3, was localized in cancer cells and macrophages along the invasive margin. These data were consistent with an active recruitment of T cells expressing CCR5 or CXCR3 into the invasive margin of colorectal cancer. With the previous clinicopathological studies showing a favorable prognostic impact of T cell infiltration in colorectal cancer, our study supports the occurrence of a certain level of Th1-shifted cellular immune responses in human colorectal cancer.
... It has been shown that in vitro, CD8 + T-IEL are able to spontaneously lyse colon carcinoma cell lines [6]. Around 20% of the stromal, but almost none of the intraepithelial CD8 + T cells express perforin in Dukes' A carcinomas, decreasing to just 3% in the stroma in Dukes' C cases [7]. Carcinoma-infiltrating Vd1 + T cells also lyse colon carcinoma cell lines [8], recognizing MICA and MICB on the tumour cells [9], and it has been shown in mice that Va14 + NK T cells lyse tumour cells [10]. ...
... The factors driving the CD8 + T cell proliferation in colorectal carcinomas are not known but are likely to be the tumour cells themselves. As the CD8 + IEL are perforin negative [7], they may have a regulatory role, rather than a cytotoxic function. The CD4 + T cells also proliferate more when in contact with the tumour. ...
We have investigated the proliferation rates of T-cell subsets in colorectal carcinomas using immunohistochemistry. It was found that the tumour-infiltrating T cells in contact with the tumour cells have a significantly higher frequency of proliferation than those in the stroma. In particular, the CD8+ intraepithelial lymphocytes (T-IEL) within the tumours have a significantly higher frequency of proliferation in comparison with CD8+ T cells in the stromal compartment or in any normal mucosal lymphoid tissues. It is possible that the proliferation of the CD8+ T-IEL may be driven by self-antigens expressed on the tumour cells. The proportion of CD3+ CD7- T cells is increased within carcinomas compared with the normal colon, and a population of CD57+ T cells was observed which is absent from the normal colon. It is possible that these phenotypes are acquired in situ due to repeated stimulation of the T cells by tumour antigens. Intact colorectal carcinoma explants were cultured, and the presence of tumour-infiltrating T cells analysed after 3 days of culture in isolation from the systemic compartments. CD3+ T cells were proliferating (at a low rate) within the explants after 3 days of culture, indicating that they may be sustained by factors present in the tumour microenvironment.
... Beim Menschen bewirkt die fehlerhafte Expression dieses Moleküls eine massive Proliferation und Infiltration von Lymphozyten [37]. Untersuchungen humaner TIL legen zudem die Vermutung nahe, dass eine geringe Anzahl Perforin-positiver Zellen im Tumor mit einem aggressiven Wachstum der Karzinome korreliert [41][42][43]. Die Expression von Perforin in TIL des RCC ist allerdings bisher wenig untersucht worden. ...
München, Univ., Diss., 2004 (Nicht für den Austausch).
All living organisms have a defense mechanism, the immune system, that protects them from potentially harmful invaders. It has become apparent that in humans the defense mechanisms of the immunologic system against microorganisms is but a part of a broader reactivity concerned with the recognition of self and non-self. Using this mechanism of recognition, the human immune system provides for the surveillance and elimination of native cells “gone wild” or that undergo malignant transformation. Malignant transformation of normal cells poses an inherent danger to the survival of the organism and, therefore, raises a threat to the species. The immune system acts in response to antigens present on the surface of transformed or neoplastic cells, but not on normal cells. As a result, the aberrant cells are destroyed before they can proliferate. Cancer results when an immune response fails to perform appropriately or adequately. Thus, immunity has come to mean a specific protective response with which the host maintains the constancy of its internal environment when confronted with substances that are recognized as foreign, be they generated from within the host or introduced from the external environment.
Molecular metastases are precursors of postoperative recurrence, detected by molecular-biological tools. Chemokines and their receptors contribute to dissemination and local immune recognition. A strong expression of the chemokine receptor CCR5 is associated with non-metastatic colorectal cancer and increased CD8+ T-cell infiltration. The aim of this study was to analyze whether CCR5 expression correlates with the presence of hepatic molecular metastases (MM).
Ninety-three patients undergoing elective surgery for colorectal cancer were assessed. The K-ras mutation status was defined by PCR-RFLP, and the CCR5 expression status was analyzed by CCR5-specific reverse transcription (RT-PCR) analysis. Liver biopsy samples had been intra-operatively taken to screen for MM. MM were detected by K-ras-specific PCR-RFLP and nested CK20/GCC RT-PCR. Prevalence of MM was correlated with CCR5 expression status.
Human colorectal cancer harboured K-ras mutations in 53% (codon 12: 47%; codon 13: 6%) of cases. Among K-ras mutants, MM were detected in 27-53% of patients, dependent on the technique applied (K-ras-specific PCR-RFLP assay vs. nested CK20/GCC RT-PCR approach (P = 0.004)). CCR5 expression of K-ras mutants ranged from absent (23/49: 47%), weak (17/49: 35%), intermediate (4/49: 8%) to strong (5/49: 10%). MM were found in 30% of CCR5 negative and in 23% of CCR5 positive cancer patients by the K-ras-specific PCR-RFLP assay. The nested CK20/GCC RT-PCR assay detected MM in 87% of CCR5 negative and in 27% of CCR5 positive colorectal cancer patients (P = 0.00002).
Thus, CCR5 expression of the primary cancer might be a valuable biomarker indicating the absence of hepatic molecular metastases.
Chemokines and their receptors have been proposed to distinctly contribute to tumor growth, dissemination, and local immune escape. The aim of this study was to evaluate the relevance of the chemokine receptor CCR5 expression for the progression of human colorectal cancer.
CCR5 expression was assessed by RT-PCR analysis in 103 colorectal cancer patients. Intensity of CCR5 expression was correlated with both tumor and patient characteristics. Infiltration of tumor margins with CD8(+) T cells in the context of CCR5 expression was analyzed by immunohistochemistry in additional 18 colorectal cancer specimens.
Human colorectal cancer revealed variable intensities of CCR5 expression ranging from absent (48/103: 47%), weak (30/103: 29%), intermediate (13/103: 13%), to strong (12/103: 12%). Absent or weak CCR5 expression was significantly associated with advanced UICC stages (P=0.02) and lymphatic metastasis (P=0.05). In addition, CCR5 expression positively correlated with CD8(+) T-cell infiltration in tumor margins (P=0.001).
In summary, intermediate and strong CCR5 expression was significantly associated with nonmetastatic colorectal cancer and increased CD8(+) T-cell infiltration.
