Enhancement of T cell recruitment and infiltration into tumours.

Studies have documented that cancer patients with tumours which are highly infiltrated with cytotoxic T lymphocytes show enhanced survival rates. The ultimate goal of cancer immunotherapy is to elicit high-avidity tumour-specific T cells to migrate and kill malignant tumours. Novel antibody therapies such as ipilumimab (a cytotoxic T lymphocyte antigen-4 blocking antibody) show enhanced T cell infiltration into the tumour tissue and increased survival. More conventional therapies such as chemotherapy or anti-angiogenic therapy and recent therapies with oncolytic viruses have been shown to alter the tumour microenvironment and thereby lead to enhanced T cell infiltration. Understanding the mechanisms involved in the migration of high-avidity tumour-specific T cells into tumours will support and provide solutions for the optimization of therapeutic options in cancer immunotherapy.

The chemokine, CXCL12, is an independent predictor of poor survival in ovarian cancer.

BACKGROUND: The chemokine CXCL12 and its cognate receptor, CXCR4, have been implicated in numerous tumour types where expression promotes tumour growth, angiogenesis, metastasis and suppresses tumour immunity. METHODS: Using a tissue microarray of 289 primary ovarian cancers coupled to a comprehensive database of clinicopathological variables, the expression of CXCL12 and CXCR4 was assessed by immunohistochemistry and its impact in terms of survival and clinicopathological variables was determined. RESULTS: Patients whose tumours expressed high levels of CXCL12 had significantly poorer survival (P=0.026) than patients whose tumours failed to produce this chemokine. Lack of CXCL12 expression within tumours was associated with a 51-month survival advantage for patients when compared with patients whose tumours expressed high levels of CXCL12. FIGO stage, adjuvant chemotherapy and the absence of macroscopic disease after surgery were all shown to predict prognosis independently of each other in this cohort of patients. CXCL12 was independently predictive of prognosis on multivariate analysis (P=0.016). There was no correlation between CXCL12 and any clinicopathological variable. CONCLUSION: The chemokine CXCL12 is an independent predictor of poor survival in ovarian cancer. High expression of CXCL12 was seen in only 20% of the tumours, suggesting a role for anti-CXCL12/CXCR4 therapy in the management of these patients.

Development of cancer vaccines to activate cytotoxic T lymphocytes.

Cancer vaccines have been shown to stimulate cytotoxic T lymphocyte (CTL) responses in a variety of cancer patients. However, the response is often of low frequency and moderate avidity, and does not result in objective clinical responses. This is related to the target antigens, which are usually over-expressed self-antigens that elicit tolerogenic and regulatory immune responses, resulting in deletion or inactivation of high-avidity T cells. Although moderate-avidity T cells can be efficient killers, tumours are often poor targets as they express a variety of molecules to protect them from cell-mediated immunity. Adoptive transfer of large numbers of high-avidity T cells has been shown to induce regression of bulky disease, proving that immune responses can effectively eradicate tumours. New approaches that target activated dendritic cells in vivo, resulting in cross-presentation of CTL epitopes and release of cytokines that suppress regulatory T cells, have resulted in the production of T cells with sufficient avidity to kill tumour target cells. These approaches in combination with regimes, such as cytokine therapy, chemotherapy or radiotherapy, that modulate effector costimulatory expression on tumour targets may result in more effective second-generation cancer vaccines.

Depressed proliferative responses by peripheral blood mononuclear cells from early arthritis patients to mycobacterial heat shock protein 60.

OBJECTIVES: T-cell responses to mycobacterial heat shock protein 60 (M.hsp60) have been implicated in the pathogenesis of adjuvant arthritis, but whether they play a role in rheumatoid arthritis (RA) is undefined. We therefore examined T-cell responses to M.hsp60 and to other recall antigens in a cohort of patients with early RA and in healthy controls. METHODS: In vitro peripheral blood mononuclear cells' (PBMC) proliferative responses to antigen were measured by [3H]thymidine incorporation, and results correlated with clinical and laboratory features of disease. RESULTS: Whereas responses to the recall antigens tetanus toxin and purified protein derivative (PPD) were equivalent in the two groups, responses to both M.hsp60 and the Escherichia coli hsp60 were lower in the RA patients. These results could not be explained by either the higher prevalence of HLA-DR4 in the RA group, or the disease severity of the patients. CONCLUSION: In the light of results from the adjuvant arthritis model which suggest that arthritis may be ameliorated by the actions of an hsp60-reactive T-cell population, the lack of response to M.hsp60 in RA could contribute to disease persistence.

T cell responses to heat-shock protein 60: differential responses by CD4+ T cell subsets according to their expression of CD45 isotypes.

We demonstrate that human T lymphocytes proliferate in vitro to highly purified human heat-shock protein 60 (Hu.hsp60). The response to this self Ag was confined to the CD45RA+ RO- T cell subset, with minimal responses by adult CD45RA- RO+ T cells. Experiments using keyhole limpet hemocyanin as a prototypic novel Ag, or tetanus toxoid as a recall Ag, were consistent with the notion that CD45RA+ RO- and CD45RA- RO+ T cell subsets can be designated as naive and memory cells, respectively; thus, responses to Hu.hsp60 were confined to the putative naive subset. In contrast, both CD45RA+ RO- and CD45RA- RO+ T cell populations proliferated to bacterial hsp60 from Mycobacterium leprae, Escherichia coli, or Chlamydia trachomatis. However, only CD45RA- RO+ (memory) T cells responded to a mycobacterial hsp60-derived peptide previously defined as a major bacteria-specific epitope. Experiments with cord blood T cells, which are CD45RA+ RO- and can be considered truly naive, showed that the peptide could elicit responses from naive T cells in vitro; cord blood cells also responded to Hu.hsp60. Since bacterial hsp60 Ags contain both conserved and nonconserved epitopes, we speculate that in vivo challenge with bacterial hsp60 will activate T cells capable of seeing either type of epitope, but only those that see nonconserved epitopes maintain the CD45RA- RO+ memory phenotype. However, T cells recognizing conserved epitopes, while not apparently being recruited to the memory pool, may nevertheless play a role in immunoregulation, particularly in the context of inflammation, when expression of Hu.hsp60 is increased.

In vitro responses of human CD45R0brightRA- and CD45R0-RAbright T cell subsets and their relationship to memory and naive T cells.

The cellular basis of immunological memory, particularly with respect to T cells is not understood. In humans, monoclonal antibodies to CD45 have been used to identify memory (CD45R0) and naive (CD45RA) T cells. However, this identification has been called into question by various studies which suggest that high molecular weight CD45 isoforms may be re-expressed by previously activated cells. In the present study, using cultures which supported responses of naive T cells, we examined the responses of purified CD45R0brightRA- or CD45R0(-)-RAbright T cell subsets. The former subset was found to respond preferentially to recall antigens with minimal responses apparent to neo-(or non-recall)-antigens. The inverse pattern was found for CD45R0-RAbright T cells, which converted to CD45R0brightRA- after stimulation with a neo-antigen. Moreover, the two populations of T cells exhibited distinct response kinetics with a faster response evident from the CD45R0brightRA- T cells compared to the CD45R0-RAbright subset. The poor responses of CD45R0-RAbright T cells to recall antigens compared to neo-antigens suggests that this putative naive population is specifically depleted of reactive T cells following an encounter with antigen. We propose that T cell priming results in the stimulation of many CD45R0-RAbright T cells with various T cell receptor specificities from which memory T cells are selected for survival. If re-expression of higher molecular weight isoforms does occur in humans in vivo, our results suggest that R0 expression would be retained (CD45R0+RA+). Alternatively, if primed CD45R0-RAbright T cells exist, they are not prevalent in peripheral blood and thus may be sequestered within lymphoid tissues. Our data support the view that in human peripheral blood, CD45R0bright and CD45RAbright expression identify memory and naive CD4+ T cells, respectively.