The expression of MHC class II molecules on murine breast tumors delays T-cell exhaustion, expands the T-cell repertoire, and slows tumor growth
The expression of MHC class II molecules (MHCII) on tumor cells correlates with survival and responsiveness to immunotherapy. However, the mechanisms underlying these observations are poorly defined. Using a murine breast tumor line, we showed that MHCII-expressing tumors grew more slowly than controls and recruited more functional CD4+ and CD8+ T cells. In addition, MHCII-expressing tumors contained more TCR clonotypes expanded to a larger degree than control tumors. Functional CD8+ T cells in tumors depended on CD4+ T cells. However, both CD4+ and CD8+ T cells eventually became exhausted, even in MHCII-expressing tumors. Treatment with anti-CTLA4, but not anti-PD-1 or anti-TIM-3, promoted complete eradication of MHCII-expressing tumors. These results suggest tumor cell expression of MHCII facilitates the local activation of CD4+ T cells, indirectly helps the activation and expansion of CD8+ T cells, and, in combination with the appropriate checkpoint inhibitor, promotes tumor regression.
KeywordsBreast cancer MHC class II T-cell exhaustion TCR repertoire
Human class II transcriptional activator
MHC class II
Murine leukemia virus
The authors would like to thank Uma Mudunuru and Scott Simpler for animal husbandry, Eddy Yang and Debbie Della Manna of the NanoString Laboratory and Enid Keyser of the Comprehensive Flow Cytometry Core for lending respective expertise. The TS/A murine mammary adenocarcinoma cell line was provided by Roberto S. Accolla, Department of Clinical and Biological Sciences, University of Insubria, Italy.
TRM designed, performed, and interpreted experiments, and wrote the manuscript. ML designed, performed, and interpreted experiments. DS performed TCR repertoire sequencing and analysis. SJC performed TCR repertoire sequencing and analysis. ML designed, performed, and interpreted experiments. SM-P designed, performed, and interpreted experiments. RCA designed and interpreted experiments and edited the manuscript. DJB designed and interpreted experiments and edited the manuscript. AF designed and interpreted experiments and edited the manuscript. TDR designed and interpreted experiments and edited the manuscript.
This work was supported by the University of Alabama at Birmingham Comprehensive Cancer Center (P30 CA013148), National Institutes of Health Grant CA216234, and by the Breast Cancer Research Foundation of Alabama.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
All procedures involving animals were performed in accordance with the guidelines of the National Research Council (United States) Committee for the Update of the Guide for the Care and Use of Laboratory Animals and were approved by the University of Alabama at Birmingham Institutional Animal Care and Use Committee (IACUC) in protocol 09854.
Research involving human participants and animals
BALB/c mice were purchased from Charles River Laboratories International, Inc. BALB/c.scid mice (CBySmn.CB17-PrkdcscidIJ) were purchased from The Jackson Laboratory.
Cell line authentication
TS/A cells were obtained at passage 22 and passaged 2 times prior to freezing archival samples. TS/A cells were authenticated by assessing MHC haplotype via flow cytometry and by detection of antigens from murine leukemia virus. Transfected and control TS/A cells were also confirmed by gene expression of MHCII pathway gene products using nanostring assay and Western blot. Both cell lines tested negative for mycoplasma (and 13 other mouse pathogens) via PCR performed by Charles River Research Animal Diagnostic Services on June 2015.
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