Anti-PD-1 increases the clonality and activity of tumor infiltrating antigen specific T cells induced by a potent immune therapy consisting of vaccine and metronomic cyclophosphamide.
Anti-PD-1 increases the clonality and activity of tumor infiltrating antigen specific T cells induced by a potent immune therapy consisting of vaccine and metronomic cyclophosphamide.
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DOI：
10.1186/s40425-016-0169-2
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发表时间：
2016
影响因子：
10.9
通讯作者：
Mansour M
中科院分区：
文献类型：
--
作者：
Weir GM;Hrytsenko O;Quinton T;Berinstein NL;Stanford MM;Mansour M
未来的癌症免疫治疗将结合多种治疗方法，通过协同、多模式机制对癌症抗原产生功能性免疫反应。在这项研究中，我们探索了三种不同的免疫疗法的组合：一类限制性肽基癌症疫苗，节拍器环磷酰胺（mCPA）和抗pd -1治疗在表达HPV16 E7 （C3）的小鼠肿瘤模型中的作用。小鼠皮下植入C3肿瘤。荷瘤小鼠以mCPA （20 mg/kg/d PO）连续治疗7天，每隔2周接种一次DepoVax （DPX）佐剂平台配制的HPV16 E749-57肽抗原，每次接种后给予抗pd -1 （200 μg/剂量IP）。通过跟踪肿瘤生长和生存来衡量疗效。采用IFN-γ ELISpot检测脾脏、疫苗引流淋巴结和肿瘤引流淋巴结的免疫原性。采用流式细胞术检测CD8α+肽特异性T细胞浸润情况，RT-qPCR检测细胞毒性蛋白浸润情况。利用基因组DNA对肿瘤浸润T细胞进行TCRβ测序，检测肿瘤浸润T细胞的克隆性。未经治疗的C3肿瘤体内PD-L1表达较低，单独抗pd -1治疗对肿瘤生长没有保护作用。DPX/mCPA治疗可延缓肿瘤生长，DPX/mCPA/anti-PD-1三联治疗可长期控制肿瘤。我们发现，与DPX/mCPA组相比，通过IFN-γ ELISpot检测，DPX/mCPA/抗pd -1治疗增强了脾脏中检测到的全身抗原特异性免疫反应，但肿瘤引流淋巴结的免疫反应没有增加。虽然没有检测到抗原特异性CD8α+ TILs的增加，但与单独使用抗pd -1或DPX/mCPA处理的小鼠相比，DPX/mCPA/抗pd -1处理的小鼠肿瘤微环境中细胞毒性基因的表达有增加的趋势，并且克隆性增加。利用抗原特异性CD8α+ T细胞克隆文库，我们发现抗原特异性克隆在DPX/mCPA/抗pd -1处理组中扩增更为频繁。这些结果表明，结合有效的靶向T细胞激活免疫疗法可以提高抗pd -1的疗效。本文的在线版本（doi:10.1186/s40425-016-0169-2）包含补充材料，可供授权用户使用。
Future cancer immunotherapies will combine multiple treatments to generate functional immune responses to cancer antigens through synergistic, multi-modal mechanisms. In this study we explored the combination of three distinct immunotherapies: a class I restricted peptide-based cancer vaccine, metronomic cyclophosphamide (mCPA) and anti-PD-1 treatment in a murine tumor model expressing HPV16 E7 (C3). Mice were implanted with C3 tumors subcutaneously. Tumor bearing mice were treated with mCPA (20 mg/kg/day PO) for seven continuous days on alternating weeks, vaccinated with HPV16 E749-57 peptide antigen formulated in the DepoVax (DPX) adjuvanting platform every second week, and administered anti-PD-1 (200 μg/dose IP) after each vaccination. Efficacy was measured by following tumor growth and survival. Immunogenicity was measured by IFN-γ ELISpot of spleen, vaccine draining lymph nodes and tumor draining lymph nodes. Tumor infiltration was measured by flow cytometry for CD8α+ peptide-specific T cells and RT-qPCR for cytotoxic proteins. The clonality of tumor infiltrating T cells was measured by TCRβ sequencing using genomic DNA. Untreated C3 tumors had low expression of PD-L1 in vivo and anti-PD-1 therapy alone provided no protection from tumor growth. Treatment with DPX/mCPA could delay tumor growth, and tri-therapy with DPX/mCPA/anti-PD-1 provided long-term control of tumors. We found that treatment with DPX/mCPA/anti-PD-1 enhanced systemic antigen-specific immune responses detected in the spleen as determined by IFN-γ ELISpot compared to those in the DPX/mCPA group, but immune responses in tumor-draining lymph nodes were not increased. Although no increases in antigen-specific CD8α+ TILs could be detected, there was a trend for increased expression of cytotoxic genes within the tumor microenvironment as well as an increase in clonality in mice treated with DPX/mCPA/anti-PD-1 compared to those with anti-PD-1 alone or DPX/mCPA. Using a library of antigen-specific CD8α+ T cell clones, we found that antigen-specific clones were more frequently expanded in the DPX/mCPA/anti-PD-1 treated group. These results demonstrate how the efficacy of anti-PD-1 may be improved by combination with a potent and targeted T cell activating immune therapy. The online version of this article (doi:10.1186/s40425-016-0169-2) contains supplementary material, which is available to authorized users.
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影响因子：
10.1
作者：
Binder DC;Engels B;Arina A;Yu P;Slauch JM;Fu YX;Karrison T;Burnette B;Idel C;Zhao M;Hoffman RM;Munn DH;Rowley DA;Schreiber H
通讯作者：
Schreiber H
影响因子：
11.2
作者：
Karyampudi L;Lamichhane P;Scheid AD;Kalli KR;Shreeder B;Krempski JW;Behrens MD;Knutson KL
通讯作者：
Knutson KL
影响因子：
5.8
作者：
Ellebaek, Eva;Engell-Noerregaard, Lotte;Svane, Inge Marie
通讯作者：
Svane, Inge Marie
影响因子：
5.4
作者：
FELTKAMP, MCW;SMITS, HL;KAST, WM
通讯作者：
KAST, WM
影响因子：
30.5
作者：
Blackburn, Shawn D.;Shin, Haina;Haining, W. Nicholas;Zou, Tao;Workman, Creg J.;Polley, Antonio;Betts, Michael R.;Freeman, Gordon J.;Vignali, Dario A. A.;Wherry, E. John
通讯作者：
Wherry, E. John