Cancer Immunology, Immunotherapy

, Volume 68, Issue 2, pp 175–188 | Cite as

The expression of MHC class II molecules on murine breast tumors delays T-cell exhaustion, expands the T-cell repertoire, and slows tumor growth

  • Tyler R. McCaw
  • Mei Li
  • Dmytro Starenki
  • Sara J. Cooper
  • Mingyong Liu
  • Selene Meza-Perez
  • Rebecca C. Arend
  • Donald J. Buchsbaum
  • Andres Forero
  • Troy D. RandallEmail author
Original Article


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.


Breast cancer MHC class II T-cell exhaustion TCR repertoire 



Glycoprotein 70


Granzyme B


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.

Author contributions

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.

Ethical approval

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.

Supplementary material

262_2018_2262_MOESM1_ESM.pdf (2.6 mb)
Supplementary material 1 (PDF 2627 KB)


  1. 1.
    Pages F, Galon J, Dieu-Nosjean MC, Tartour E, Sautes-Fridman C, Fridman WH (2010) Immune infiltration in human tumors: a prognostic factor that should not be ignored. Oncogene 29(8):1093–1102. CrossRefGoogle Scholar
  2. 2.
    Schreiber RD, Old LJ, Smyth MJ (2011) Cancer immunoediting-integrating immunity’s roles in cancer suppression and promotion. Science 331(6024):1565–1570. CrossRefGoogle Scholar
  3. 3.
    Topalian SL, Drake CG, Pardoll DM (2015) Immune checkpoint blockade: a common denominator approach to cancer therapy. Cancer Cell 27(4):450–461. CrossRefGoogle Scholar
  4. 4.
    Prieto PA, Yang JC, Sherry RM, Hughes MS, Kammula US, White DE et al (2012) CTLA-4 blockade with ipilimumab: long-term follow-up of 177 patients with metastatic melanoma. Clin Cancer Res 18(7):2039–2047. CrossRefGoogle Scholar
  5. 5.
    Robert C, Long GV, Brady B, Dutriaux C, Maio M, Mortier L et al (2015) Nivolumab in previously untreated melanoma without BRAF mutation. N Engl J Med 372(4):320–330. CrossRefGoogle Scholar
  6. 6.
    Rizvi NA, Hellmann MD, Snyder A, Kvistborg P, Makarov V, Havel JJ et al (2015) Mutational landscape determines sensitivity to PD-1 blockade in NSCLC. Science 348:124–128. CrossRefGoogle Scholar
  7. 7.
    Snyder A, Makarov V, Merghoub T, Yuan J, Zaretsky JM, Desrichard A et al (2014) Genetic basis for clinical response to CTLA-4 blockade in melanoma. N Engl J Med 371(23):2189–2199. CrossRefGoogle Scholar
  8. 8.
    Johnson DB, Estrada MV, Salgado R, Sanchez V, Doxie DB, Opalenik SR et al (2016) Melanoma-specific MHC-II expression represents a tumour-autonomous phenotype and predicts response to anti-PD-1/PD-L1 therapy. Nat Commun 7:10582. CrossRefGoogle Scholar
  9. 9.
    Matsushita K, Takenouchi T, Shimada H, Tomonaga T, Hayashi H, Shioya A et al (2006) Strong HLA-DR antigen expression on cancer cells relates to better prognosis of colorectal cancer patients: possible involvement of c-myc suppression by interferon-gamma in situ. Cancer Sci 97(1):57–63. CrossRefGoogle Scholar
  10. 10.
    Schonocchia G, Eppenberger-Castori S, Zlobec I, Karamitopoulou E, Arriga R, Coppola A et al (2014) HLA class II antigen expression in colorectal carcinoma tumors as a favorable prognostic marker. Neoplasia 16(1):31–42. CrossRefGoogle Scholar
  11. 11.
    Diepstra A, van Imhoff GW, Karim-Kos HE, van den Berg A, te Meerman GJ, Niens M et al (2007) HLA class II expression by Hodgkin Reed–Sternberg cells is an independent prognostic factor in classical Hodgkin’s lymphoma. J Clin Oncol 25(21):3101–3108. CrossRefGoogle Scholar
  12. 12.
    Tada K, Maeshima AM, Hiraoka N, Yamauchi N, Maruyama D, Kim SW et al (2016) Prognostic significance of HLA class I and II expression in patients with diffuse large B cell lymphoma treated with standard chemoimmunotherapy. Cancer Immunol Immunother 65(10):1213–1222. CrossRefGoogle Scholar
  13. 13.
    Callahan MJ, Nagymanyoki Z, Bonome T, Johnson ME, Litkouhi B, Sullivan EH et al (2008) Increased HLA-DMB expression in the tumor epithelium is associated with increased CTL infiltration and improved prognosis in advanced-stage serous ovarian cancer. Clin Cancer Res 14(23):7667–7673. CrossRefGoogle Scholar
  14. 14.
    Forero A, Li Y, Chen D, Grizzle WE, Updike KL, Merz ND et al (2016) Expression of the MHC class II pathway in triple-negative breast cancer tumor cells is associated with a good prognosis and infiltrating lymphocytes. Cancer Immunol Res 4(5):390–399. CrossRefGoogle Scholar
  15. 15.
    Oldford SA, Robb JD, Codner D, Gadag V, Watson PH, Drover S (2006) Tumor cell expression of HLA-DM associates with a Th1 profile and predicts improved survival in breast carcinoma patients. Int Immunol 18(11):1591–1602. CrossRefGoogle Scholar
  16. 16.
    Reith W, LeibundGut-Landmann S, Waldburger JM (2005) Regulation of MHC class II gene expression by the class II transactivator. Nat Rev Immunol 5(10):793–806. CrossRefGoogle Scholar
  17. 17.
    Cella M, Scheidegger D, Palmer-Lehmann K, Lane P, Lanzavecchia A, Alber G (1996) Ligation of CD40 on dendritic cells triggers production of high levels of IL-12 and enhances T cell stimulatory capacity- T-T help via APC activation. J Exp Med 184:747–752. CrossRefGoogle Scholar
  18. 18.
    Schoenberger SP, Toes REM, van der Voort EIH, Offringa R, Melief CJM (1998) T-cell help for cytotoxic T lymphocytes is mediated by CD40-CD40L interactions. Nature 393:480–483. CrossRefGoogle Scholar
  19. 19.
    Meazza R, Comes A, Orengo AM, Ferrini S, Accolla RS (2003) Tumor rejection by gene transfer of the MHC class II transactivator in murine mammary adenocarcinoma cells. Eur J Immunol 33(5):1183–1192. CrossRefGoogle Scholar
  20. 20.
    Mortara L, Castellani P, Meazza R, Tosi G, De Lerma Barbaro A, Procopio FA et al (2006) CIITA-induced MHC class II expression in mammary adenocarcinoma leads to a Th1 polarization of the tumor microenvironment, tumor rejection, and specific antitumor memory. Clin Cancer Res 12(11 Pt 1):3435–3443. CrossRefGoogle Scholar
  21. 21.
    Zajac AJ, Blattman JN, Murali-Krishna K, Sourdive DJD, Suresh M, Altman JD et al (1998) Viral immune evasion due to persistence of activated T cells without effector function. J Exp Med 188(12):2205–2213. CrossRefGoogle Scholar
  22. 22.
    Wherry EJ, Blattman JN, Murali-Krishna K, van der Most R, Ahmed R (2003) Viral persistence alters CD8 T-cell immunodominance and tissue distribution and results in distinct stages of functional impairment. J Virol 77(8):4911–4927. CrossRefGoogle Scholar
  23. 23.
    Bevan MJ (2004) Helping the CD8+ T-cell response. Nat Rev Immunol 4(8):595–602. CrossRefGoogle Scholar
  24. 24.
    Bourgeois C, Veiga-Fernandes H, Joret A, Rocha B, Tanchot C (2002) CD8 lethargy in the absence of CD4 help. Eur J Immunol 32:2199–2207.;2-L.CrossRefGoogle Scholar
  25. 25.
    Rosato A, Santa SD, Zoso A, Giancomelli S, Milan G, Macino B et al (2003) The cytotoxic T-lymphocyte response against a poorly immunogenic mammary adenocarcinoma is focused on a single immunodominant class I epitope derived from the gp70 env product of an endogenous retrovirus. Cancer Res 63:2158–2163Google Scholar
  26. 26.
    Dummer W, Niethammer AG, Baccala R, Lawson BR, Wagner N, Reisfeld RA et al (2002) T cell homeostatic proliferation elicits effective antitumor autoimmunity. J Clin Invest 110:185–192. CrossRefGoogle Scholar
  27. 27.
    Gattinoni L, Finkelstein SE, Klebanoff CA, Antony PA, Palmer DC, Spiess PJ et al (2005) Removal of homeostatic cytokine sinks by lymphodepletion enhances the efficacy of adoptively transferred tumor-specific CD8+ T cells. J Exp Med 202(7):907–912. CrossRefGoogle Scholar
  28. 28.
    Bennaceur K, Chapman J, Brikci-Nigassa L, Sanhadji K, Touraine JL, Portoukalian J (2008) Dendritic cells dysfunction in tumour environment. Cancer Lett 272(2):186–196. CrossRefGoogle Scholar
  29. 29.
    Gabrilovich DI, Ostrand-Rosenberg S, Bronte V (2012) Coordinated regulation of myeloid cells by tumours. Nat Rev Immunol 12(4):253–268. CrossRefGoogle Scholar
  30. 30.
    Qi L, Rojas J, Ostrand-Rosenberg S (2000) Tumor cells present MHC class II-restricted nuclear and mitochondrial antigens and are the predominant antigen presenting cells in vivo. J Immunol 165:5451–5461. CrossRefGoogle Scholar
  31. 31.
    Antony PA, Piccirillo CA, Akpinarli A, Finkelstein SE, Speiss PJ, Surman DR et al (2005) CD8+ T cell immunity against a tumor-self-antigen is augmented by CD4+ T helper cells and hindered by naturally occurring T regulatory cells. J Immunol 174:2591–2601. CrossRefGoogle Scholar
  32. 32.
    Bos R, Sherman LA (2010) CD4+ T-cell help in the tumor milieu is required for recruitment and cytolytic function of CD8+ T lymphocytes. Cancer Res 70(21):8368–8377. CrossRefGoogle Scholar
  33. 33.
    Inderberg-Suso EM, Trachsel S, Lislerud K, Rasmussen AM, Gaudernack G (2012) Widespread CD4+ T-cell reactivity to novel hTERT epitopes following vaccination of cancer patients with a single hTERT peptide GV1001. Oncoimmunology 1(5):670–686. CrossRefGoogle Scholar
  34. 34.
    Prickett TD, Crystal JS, Cohen CJ, Pasetto A, Parkhurst MR, Gartner JJ et al (2016) Durable complete response from metastatic melanoma after transfer of autologous T cells recognizing 10 mutated tumor antigens. Cancer Immunol Res 4(8):669–678. CrossRefGoogle Scholar
  35. 35.
    Prlic M, Blazar BR, Khoruts A, Zell T, Jameson SC (2001) Homeostatic expansion occurs independently of costimulatory signals. J Immunol 167:5664–5668. CrossRefGoogle Scholar
  36. 36.
    Voehringer D, Liang H, Locksley RM (2008) Homeostasis and effector function of lymphopenia-induced memory like T cells in constitutively T cell depleted mice. J Immunol 180:4742–4753. CrossRefGoogle Scholar
  37. 37.
    Wherry EJ, Kurachi M (2015) Molecular and cellular insights into T cell exhaustion. Nat Rev Immunol 15(8):486–499. CrossRefGoogle Scholar
  38. 38.
    Spranger S, Spaapen RM, Zha Y, Williams J, Meng Y, Ha TT et al (2013) Up-regulation of PD-L1, IDO, and Tregs in the melanoma tumor microenvironment is driven by CD8+ T cells. Sci Transl Med 5(200):1–10. CrossRefGoogle Scholar
  39. 39.
    Benci JL, Xu B, Qiu Y, Wu TJ, Dada H, Twyman-Saint Victor C et al (2016) Tumor interferon signaling regulates a multigenic resistance program to immune checkpoint blockade. Cell 167(6):1540–1554 e12. CrossRefGoogle Scholar
  40. 40.
    Taube JM, Klein A, Brahmer JR, Xu H, Pan X, Kim JH et al (2014) Association of PD-1, PD-1 ligands, and other features of the tumor immune microenvironment with response to anti-PD-1 therapy. Clin Cancer Res 20(19):5064–5074. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Division of Clinical Immunology and Rheumatology, Department of MedicineUniversity of Alabama at BirminghamBirminghamUSA
  2. 2.Department of Radiation OncologyUniversity of Alabama at BirminghamBirminghamUSA
  3. 3.HudsonAlpha Institute for BiotechnologyHuntsvilleUSA
  4. 4.Department of Obstetrics and GynecologyUniversity of Alabama at BirminghamBirminghamUSA
  5. 5.Division of Hematology and Oncology, Department of MedicineUniversity of Alabama at BirminghamBirminghamUSA

Personalised recommendations