Functional Tumor Infiltrating TH1 and TH2 Effectors in Large Early-Stage Cervical Cancer Are Suppressed by Regulatory T Cells

Objective: Analysis of tumor-infiltrating lymphocytes (TILs) is one of the cornerstones for the understanding of immune responses prevailing in the tumor microenvironment. We studied TILs from squamous cell carcinoma of the cervix ex vivo without proliferating them in vitro before analysis. Methods: Whereas TILs were magnetic activated cell separation enriched and flow sorted into CD4(+) CD25(hi) (regulatory T cells [Tregs]), CD4(+) CD25(int) (effector T cells [Teffs]) were directly purified by flow cytometry, and both these subsets were characterized phenotypically and functionally. Tissue sections were probed for interleukin 4 (IL-4) and interferon gamma. Results: Effector T cells constitutively expressed both interferon gamma and IL-4 prototypical cytokines of T(H)1 and T(H)2, respectively, and were able to proliferate and secrete higher quantities of both cytokines in response to anti-CD3/anti-CD28 and autologous tumor lysates. Only 53% of cervical cancer Tregs were FOXP3(+), elaborated transforming growth factor beta 1, and IL-10 and were able to inhibit both T helper subsets. Conclusions: Intratumoral Teffs represented functionally active subsets of both T(H)1 andT(H)2 that were not anergic but were suppressed by multiple Treg subsets, which comprised FOXP3 + Tregs and Tregs secreting transforming growth factor beta 1 and IL-10. These results imply that the microenvironment of cervical carcinomas harbored both T(H)1 and T(H)2 subsets of CD4(+) Teffs that were functionally active but were perhaps unable to perform because of the overpowering effect of Tregs.