Elsevier

Lung Cancer

Volume 47, Issue 3, March 2005, Pages 361-371
Lung Cancer

Effector, memory and naïve CD8+ T cells in peripheral blood and pleural effusion from lung adenocarcinoma patients

https://doi.org/10.1016/j.lungcan.2004.07.046Get rights and content

Summary

The proportions of naïve, memory and effector CD8+ T cells in peripheral blood and pleural effusion from lung adenocarcinoma patients were studied. CD8+ T subsets were identified by using a combination of the following antibodies: anti-CD45RA, anti-CD45RO, anti-CD27 and anti-CD28, as well as antibodies to other markers. Fas-positive cells were determined in each CD8+ T subset. Also, the intracellular cytokine patterns of CD4+ and CD8+ lymphocytes from pleural effusion were analysed. In naïve, memory and effector CD8+ T subsets no significant differences were observed in peripheral blood between healthy donors and cancer patients. In contrast, a high proportion of cells with memory phenotype (CD45RA−CD45RO+CD27+CD28+) and a low proportion of cells with effector phenotype (CD45RA+CD45RO−CD27−CD28−) were found in pleural effusion with respect to peripheral blood (P < 0.001). The altered proportions of CD8+ T subsets in pleural effusion were not mediated by type 2 cytokines produced by CD4+ or CD8+ lymphocytes.

In the effector CD8+ T subset, from peripheral blood as well as from pleural effusion, a low percentage of perforin-expressing cells was observed compared to granzyme A-expressing cells. Additionally, a high percentage of naïve CD8+ T cells expressing Fas was found. Our data suggest that: (i) terminal-differentiation process of CD8+ T cells is blocked, and (ii) early Fas-expression in CD8+ T cells, which was reflected even in peripheral blood, may lead to apoptosis of naïve cells when they reach the effector stage. All these processes may contribute to the inadequate antitumour immune response found in lung carcinoma patients.

Introduction

CD8+ T cells represent a major arm of cell-mediated immune response. These cells recognise and lyse cells carrying non-self epitopes, such as virus-infected cells, grafted tissues or tumour cells. Although genetic, phenotypic and functional studies have been done to assess the pathways of CD8+ T cell differentiation, these have not been fully understood [1], [2], [3], [4]. Whereas the pattern of CD8+ T cell differentiation in the mouse follows the canonical model naïve  effector  memory, in the human several studies [2], [3], [5] have shown the pattern naïve  memory  effector.

The expression of CD45RA or CD45RO isoforms are used to identify naïve and memory CD8+ T cells, respectively. Some reports indicate that within the naïve CD8+CD45RA+ T cell compartment, two distinct subsets can be identified according to CD27 or CD28 expression [1], [2], [3]. Based on these studies, three distinct stages on the CD8 T cell differentiation process can be recognised. (1) Naïve CD8+ T cells are described as CD45RA+CD45RO−CD27+CD28+ cells; this subset produces IL-2 after stimulation, and expresses neither granzymes nor perforin [1], [3]. (2) Memory CD8+ T cells, with CD45RA−CD45RO+CD27+CD28+ phenotype, produce several cytokines after stimulation, and express granzymes but not perforin or express it at a low level [1], [3]. (3) Terminally differentiated (effector) CD8+ T cells, described as CD45RA+CD45RO−CD27−CD28− cells, express granzymes and perforin at a high level [1], [3], [6].

Lung carcinoma is the most common fatal cancer worldwide and adenocarcinoma is the histological type of highest incidence. Pleural effusion is a common manifestation of metastatic lung tumours. Large numbers of pleural effusion mononuclear cells (PEMC) and tumour cells are present in the effusion [7]. Some studies have shown that PEMC exhibit several functional defects [8], [9]. In addition, previous clinical trials evaluating specific and non-specific immune stimulation for the treatment of lung cancer associated to pleural effusion have shown poor results [10], [11].

To our knowledge, there are no reports regarding CD8+ T subsets in lung carcinoma. The aim of this study was to evaluate the frequency of CD8+ T subsets in peripheral blood and pleural effusion from lung adenocarcinomas; in addition, molecules involved in the cytolytic machinery were analysed. Our results show a higher proportion of CD8+ T cells with a memory phenotype and a lower proportion of cells with an effector phenotype in pleural effusion compared to peripheral blood. We found that this phenomenon is not associated to the production of type 2 cytokines by CD4+ or CD8+ lymphocytes; but it might depend on: (1) a block in the terminal differentiation pathway of CD8+ T cells, (2) the early Fas-expression in the naïve CD8+ T subset, which may lead to an increased susceptibility to apoptosis when the naïve cells reach the terminal differentiation stage, or (3) both processes.

Section snippets

Population studied

The population consisted of 13 patients with pleural effusion caused by lung adenocarcinoma. The diagnosis was established by histological examination of pleural biopsy or cytological observation of malignant cells in pleural effusion according to WHO criteria [12]. The malignancy stage was evaluated according to the UICC TNM classification [13] and was as follows: four patients were included in stage IIIb and nine patients in stage IV. All patients were treatment-free. Median age of this group

Proportion of CD3+, CD4+ and CD8+ T cells

In peripheral blood, the percentages of CD3+, CD4+ and CD8+ T lymphocytes from adenocarcinoma patients were not statistically different from those of healthy donors. See Fig. 1.

In lung adenocarcinoma patients, the percentages of CD3+ T cells were significantly higher in pleural effusion (68%, range: 46–90%) than in peripheral blood (43%, range: 33–81%). The percentages of CD4+ T cells were similar in pleural effusion and peripheral blood. In contrast, the percentages of CD8+ T cells were

Discussion

In pleural effusion caused by lung adenocarcinoma, other authors [15], [17] have described an increase in the number of CD3+ and CD4+ T cells with a decrease in CD8+ T cells; our data agree with these observations. In addition, several groups [8], [9], [19] have reported the following defects in immune cells from peripheral blood and pleural effusion of patients with lung carcinoma: (a) reduced proliferation rate, (b) diminished production of some cytokines, (c) reduced capacity of cytotoxic

Conclusions

Tumour microenvironment induces alterations in the CD8+ T cell differentiation program, maintaining it in the memory stage. In contrast to the increased proportion of the memory subset, the proportion of the effector subset is greatly reduced. In addition, the proportion of effector cells producing perforin is also greatly reduced. In naïve cells the proportion of cells expressing Fas is increased. This might lead to apoptosis when the cells reach the effector stage. Although all these

Acknowledgements

This work was supported in part by CONACyT grant F643-M9406. Heriberto Prado-Garcia was supported by CONACyT scholarship 142871.

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