Abstract
BackgroundWe examined the phenotype and function of lymphocytes collected from the peripheral blood (PBL) and tumor (TIL) of patients with two different solid malignancies: colorectal cancer liver metastases (CRLM) and ovarian cancer (OVC).MethodsTumor and corresponding peripheral blood were collected from 16 CRLM and 22 OVC patients; immediately following resection they were processed and analyzed using a multi-color flow cytometry panel. Cytokine mRNA from purified PBL and TIL CD4+ T cells were also analyzed by qPCR.ResultsOverall, we found similar changes in the phenotypic and cytokine profiles when the TIL were compared to PBL from patients with two different malignancies. The percentage of Treg (CD4+/CD25+/FoxP3+) in PBL and TIL was similar: 8.1% versus 10.2%, respectively in CRLM patients. However, the frequency of Treg in primary OVC TIL was higher than PBL: 19.2% versus 4.5% (p <0.0001). A subpopulation of Treg expressing HLA-DR was markedly increased in TIL compared to PBL in both tumor types, CRLM: 69.0% versus 31.7% (p = 0.0002) and OVC 74.6% versus 37.0% (p <0.0001), which suggested preferential Treg activation within the tumor. The cytokine mRNA profile showed that IL-6, a cytokine known for its immunosuppressive properties through STAT3 upregulation, was increased in TIL samples in patients with OVC and CRLM. Both TIL populations also contained a significantly higher proportion of activated CD8+ T cells (HLA-DR+/CD38+) compared to PBL (CRLM: 30.2% vs 7.7%, (p = 0.0012), OVC: 57.1% vs 12.0%, (p <0.0001)).ConclusionThis study demonstrates that multi-color flow cytometry of freshly digested tumor samples reveals phenotypic differences in TIL vs PBL T cell sub-populations. The TIL composition in primary and metastatic tumors from two distinct histologies were remarkably similar, showing a greater proportion of activated/suppressive Treg (HLA-DR+, CD39+, CTLA-4+ and Helios+) and activated cytotoxic T cells (CD8+/HLA-DR+/CD38+) when compared to PBL and an increase in IL-6 mRNA from CD4 TIL.Electronic supplementary materialThe online version of this article (doi:10.1186/s40425-014-0038-9) contains supplementary material, which is available to authorized users.
Highlights
We examined the phenotype and function of lymphocytes collected from the peripheral blood (PBL) and tumor (TIL) of patients with two different solid malignancies: colorectal cancer liver metastases (CRLM) and ovarian cancer (OVC)
We found that infiltrating lymphocytes from both tumor types had a higher CD8/CD4 T cell ratio compared to PBL
While we have found an increase in activated Treg, we found that the CD4+ tumor infiltrating lymphocytes (TIL) express higher levels of IL-6 mRNA compared to CD4+ PBL
Summary
We examined the phenotype and function of lymphocytes collected from the peripheral blood (PBL) and tumor (TIL) of patients with two different solid malignancies: colorectal cancer liver metastases (CRLM) and ovarian cancer (OVC). Tumor immunotherapy has emerged as an important treatment modality for cancer patients. Melanoma and renal cell carcinoma have been described as tumors that respond to immunotherapy. Most immunologic analyses in cancer patients have been conducted on PBL or tissue sections, where samples are more obtained. It is the composition of the immune cells at the tumor site that is likely to be most important; and the tumor infiltrating lymphocytes (TIL) may be quite different from the PBL. To gain a deeper understanding of how immunotherapy could potentially affect the tumor microenvironment, it would be important to first characterize the subset distribution and phenotype of the immune cells in progressively growing tumors
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