Elsevier

European Urology

Volume 61, Issue 2, February 2012, Pages 385-394
European Urology

Kidney Cancer
T Lymphocyte Recruitment into Renal Cell Carcinoma Tissue: A Role for Chemokine Receptors CXCR3, CXCR6, CCR5, and CCR6

https://doi.org/10.1016/j.eururo.2011.10.035Get rights and content

Abstract

Background

Evidence suggests that some patients with renal cell carcinoma (RCC) respond to immunomodulatory therapies that activate T lymphocytes. A prerequisite for effective T cell therapy is efficient targeting of effector T cells to the tumour site, yet the molecular basis of T cell recruitment to RCC is unknown. Furthermore, some T cells that naturally infiltrate this cancer are regulatory T cells (Tregs) that may suppress antitumour immune responses.

Objective

Determine the mechanisms of effector and regulatory T cell recruitment to RCC to allow targeted therapy that promotes local anti-tumour immunity.

Design, setting, and participants

Tumour-infiltrating and peripheral blood T cells were collected from 70 patients undergoing nephrectomy for RCC.

Measurements

T cells were analysed by multicolour flow cytometry for expression of 19 chemokine receptors and 7 adhesion molecules. Receptors that were expressed at higher levels on tumour-infiltrating lymphocytes (TILs) compared with matched peripheral blood lymphocytes (PBLs) were analysed further for their ability to mediate migration responses in TILs and for expression of corresponding ligands in tumour tissue.

Results and limitations

Three chemokine receptors—CCR5, CXCR3, and CXCR6—were significantly overexpressed on TILs compared with matched PBLs (n = 16 cases) and were capable of promoting migration in vitro. Their corresponding ligands CCL4–5, CXCL9–11, and CXCL16 were all detected in RCC tissue. However, since they were present in all cases studied, it was not possible to correlate ligand expression with levels of T cell infiltration. Foxp3+ Tregs were enriched within TILs compared with matched PBLs and expressed high levels of CCR5, CXCR3, and CXCR6, as well as CCR6, the ligand for which (CCL20) was detectable in RCC tissue.

Conclusions

Our data support a role for CCR5, CXCR3, and CXCR6 in the selective recruitment of T cells into RCC tissue and, together with CCR6, in the recruitment of Tregs.

Introduction

Renal cell carcinoma (RCC) is a good candidate for T lymphocyte–based therapies for the following reasons. First, immune therapies are effective in some patients with RCC, as demonstrated by cure rates of ≤7% in patients treated with high-dose interleukin 2 (IL-2) [1]. Second, allogeneic stem cell transplantation can result in regression of metastatic RCC, consistent with a graft-versus-tumour effect, and is associated with a donor-derived CD8+ T cell response to a defined RCC target antigen [2], [3]. Third, RCC-specific T cells generated in vitro lyse RCC cell lines [4], [5]. Finally, rare spontaneous remissions consistent with an anti-tumour immune response have been observed [6]. These data, along with impressive clinical responses following infusions of T cells to treat other human cancers [7], highlight the potential of T cell–based therapies for RCC.

A prerequisite for effective T cell therapy is efficient targeting of tumour-specific T cells to the tumour site, yet the mechanisms whereby T cells are recruited to RCC tissue are unknown. Therefore, to identify the homing phenotype required for therapeutic T cells to access the tumour, we have analysed T cells that naturally infiltrate RCC tissue. The efficacy of effector T cells is counterbalanced by regulatory T cells (Tregs) that infiltrate many tumours and suppress local effector T cell responses [8], [9]. Tregs, identified by expression of the transcription factor Foxp3, are present in RCC, where they correlate with poor prognosis [10], [11], [12]. Thus, we sought to identify homing molecules that recruit effector T cells and Tregs to RCC to allow future therapy to promote local anti-tumour immunity by altering the balance in favour of effector cell recruitment.

Recruitment of circulating T cells into tissue is critically dependent on the activation of specific receptors by members of the family of small chemotactic cytokines, chemokines, in target tissues [13]. We have conducted the first comprehensive analysis of homing marker expression on T cell subsets in RCC tissue and identified four functional chemokine receptors involved in T cell recruitment.

Section snippets

Clinical material

Heparinised blood and sections of tumour tissue (from non-necrotic, nonhaemorrhagic central regions) were collected from 70 patients with histologically confirmed RCC removed at nephrectomy (Table 1). All patients gave written informed consent, and the Birmingham, East, North, and Solihull Research Ethics Committee approved the study. Fresh biopsy material was rinsed with RPMI 1640 (Life Technologies) to remove traces of blood, the material was disaggregated using scalpels, and the cell

Tumour-infiltrating lymphocytes express high levels of CCR5, CXCR3 and CXCR6

Initial studies showed high levels of the following homing markers on both CD4+ and CD8+ infiltrating T cell subsets from 22 RCC patients: CCR5, CXCR3, CXCR4, CXCR6, CD62L, LFA-1, PSGL-1, and VLA-4 (data not shown). We then analysed a further 16 RCC cases for receptors overexpressed on tumour-infiltrating lymphocytes (TILs) compared with matched PBLs. Three such chemokine receptors were identified—CCR5, CXCR3, and CXCR6—in which both the intensity of staining (Fig. 1A–1C) and the percentage of

Discussion

Understanding the molecular basis of T cell recruitment to RCC will aid the development of T cell–based therapies by improving delivery of effector cells to the tumour site and identifying therapeutic opportunities to block recruitment of tumour-promoting Tregs. From our comprehensive analysis of all 19 chemokine receptors and several adhesion molecules, we identified three that were overexpressed on infiltrating T cells in RCC: CCR5, CXCR3, and CXCR6. Our data on CCR5 and CXCR3 are consistent

Conclusions

CCR5, CXCR3, and CXCR6 are involved in the selective recruitment of T cells into RCC tissue. These chemokine receptors, as well as CCR6, are also involved in recruiting Tregs to the tumour site.

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