Anti-Telomerase CD4+ Th1 Immunity and Monocytic-Myeloid-Derived-Suppressor Cells Are Associated with Long-Term Efficacy Achieved by Docetaxel, Cisplatin, and 5-Fluorouracil (DCF) in Advanced Anal Squamous Cell Carcinoma: Translational Study of Epitopes-HPV01 and 02 Trials

Laurie Spehner, Stefano Kim, Angélique Vienot, Eric François, Bruno Buecher, Olivier Adotevi, Dewi Vernerey, Syrine Abdeljaoued, Aurélia Meurisse, Christophe Borg, Laurie Spehner, Stefano Kim, Angélique Vienot, Eric François, Bruno Buecher, Olivier Adotevi, Dewi Vernerey, Syrine Abdeljaoued, Aurélia Meurisse, Christophe Borg

Abstract

Docetaxel, cisplatin and 5-fluorouracil (DCF) chemotherapy regimen is highly effective in advanced anal squamous cell carcinoma (SCCA), as demonstrated by the Epitopes-HPV02 study results. Here, we analyzed the impact of DCF regimen and the prognostic value of adaptive immune responses and immunosuppressive cells in SCCA patients included in two prospective studies (Epitopes-HPV01 and HPV02). The presence of T-cell responses against Human papillomavirus (HPV)16-E6/E7 and anti-telomerase (hTERT)-antigens was measured by IFNᵧ-ELISpot. Here, we showed that HPV-adaptive immune responses are increased in SCCA patients. SCCA patients also displayed enhanced circulating TH1 T-cells restricted by hTERT. Exposition to DCF increased hTERT immunity but not HPV or common viruses immune responses. Notably, the correlation of hTERT immune responses with SCCA patients' clinical outcomes highlights that hTERT is a relevant antigen in this HPV-related disease. The influence of peripheral immunosuppressive cells was investigated by flow cytometry. While both regulatory T-cells and monocytic-myeloid-derived suppressive cells (M-MDSC) accumulated in the peripheral blood of SCCA patients, only high levels of M-MDSC were negatively correlated with hTERT adaptive immune responses and predicted poor prognosis. Altogether, our results reveal that hTERT is a relevant antigen in HPV-driven SCCA disease and that M-MDSC levels influence TH1-adaptive immune responses and patients' survival.

Keywords: Biomarkers; DCF chemotherapy; M-MDSC; advanced anal squamous cell carcinoma; hTERT antigens.

Conflict of interest statement

C.B. declared these conflicts of interest: Advisory board: MSD, Pierre Fabre, Roche, Research grant: Roche; D.V. declared these conflicts of interest: Advisory board: Haliodx, Cellprothera, Incyte, Merck Serono. All remaining authors have declared no conflicts of interest.

Figures

Figure 1
Figure 1
Frequencies and intensities of antigen-specific T-cell responses in SCCA patients. PBMC from 17 healthy donors and SCCA patients before (n = 82) and after (n = 69) DCF chemotherapy were analyzed for antigen-specific T-cell responses by IFNᵧ ELISpot assay. A-C Intensity of positive HPV16-E6 (A), HPV16-E7 (B) and hTERT (C) specific T-cell responses in healthy donors and SCCA patients before and after DCF treatment. (D) Intensity of positive antiviral T-cell responses. Healthy donors population is represented by light gray points and SCCA patients by black points. Mann Whitney U test, where ** p < 0.01, *** p < 0.001. Median with interquartile range was indicated on graphs. Only the positive intensities of specific immune responses were indicated. (E,F) Correlation between HPV16-E6- and hTERT-specific immune responses before (E) and after DCF chemotherapy (F) in SCCA patients.
Figure 2
Figure 2
Presence of antigen-specific T-cell responses are associated with the clinical outcomes of SCCA patients. (A) The absolute lymphocyte count was analyzed in peripheral blood of SCCA patients before (n = 56) and after (n = 51) DCF chemotherapy. Kaplan-Meier curve in SCCA patients before and after DCF chemotherapy according to low absolute lymphocyte count. PBMC from SCCA patients before (n = 82) and after (n = 69) DCF chemotherapy were analyzed for antigen-specific T-cell responses by IFNᵧ ELISpot assay. (B,C) Kaplan-Meier OS curve in SCCA patients according to HPV (E6 and/or E7) (B) or hTERT (C) specific T-cell responses before and after DCF chemotherapy. Log-rank test, where p < 0.05. (D) Kaplan-Meier OS curve in SCCA patients according to antiviral-specific T-cell responses before and after DCF chemotherapy.
Figure 3
Figure 3
M-MDSC levels and not Treg levels are associated with the clinical outcomes in SCCA patients. PBMC from 19 healthy donors and SCCA patients before (n = 82) and after (n = 69) DCF chemotherapy were analyzed for Treg and M-MDSC population by flow cytometry. (A) Frequencies (%) of M-MDSC. (B) Frequencies (%) of CD25+Foxp3+ expressed on CD4+ T-cells. (C) Kaplan-Meier survival curve in SCCA patients before and after DCF chemotherapy according to Treg levels. (D) Kaplan-Meier survival curve in SCCA patients before and after DCF chemotherapy according to M-MDSC levels. Log-rank test, where ** p < 0.01. (E) Kaplan-Meier curve in distribution of M-MDSC levels in SCCA patients treated by DCF chemotherapy.
Figure 4
Figure 4
M-MDSC and not Treg levels are correlated with antigen-specific immune responses. (A,B) Intensities of positive HPV16-E6-, HPV16-E7-, hTERT- and antiviral-specific T-cell responses in SCCA patients before and after DCF chemotherapy according to Treg levels (A) or M-MDSC levels (B). Mann Whitney U test, where ** p < 0.01. Red plots represent patients without progression.

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