CD24⁺ MDSC-DCs Induced by CCL5-Deficiency Showed Improved Antitumor Activity as Tumor Vaccines

Abstract

Background Dendritic cell (DC) tumor vaccine has been extensively utilized in preclinical and clinical studies; however, this technique has encountered many difficulties, particularly in late-stage tumor patients. For those, ex vivo-induced DCs are actuallymyeloid-derived suppressive cells-derived DCs (MDSC-DCs). MDSCs with immunosuppressive activity, but not monocytes, became the major DC precursor. Thus, how to enhance antitumor activity of MDSC-DCs is urgent need to address.
Methods We utilized 4T1 and MC38 tumor-bearing both wildtype and CC chemokine ligand 5^-/- (CCL5^-/-) mice as animal models. MDSC-DCs were induced from splenocytes of these mice by granulocyte macrophage-colony stimulating factor/interleukin-4 with or without all-trans-retinoic acid (ATRA) in vitro for 7 days, then incubated with tumor-cell-lysis to treat mouse models for total three doses. For human MDSC-DCs, peripheral bloods from colorectal cancer patients were induced in vitro as murine cells with or without T- lymphocytes depletion to get rid of CCL5.
Results Flow cytometry analysis showed that MDSCs from CCL5 ^-/- mice could be induced into a new type of CD24⁺ MDSC-DCs in the presence of ATRA, which had more antitumor activity than control. Antibody blocking and adoptive transfer experiments demonstrated that downregulation of regulatory T cells (Tregs) mediated the inhibition of CD24⁺ MDSC-DCs on tumor growth. Mechanically, CD24⁺ MDSC-DCs inhibited Tregs' polarization by secreting cytokine or coactivators' expression. What's important, decreasing CCL5 protein levels by T- lymphocytes depletion during both murine and human MDSC-DCs in vitro induction could also acquire CD24⁺ MDSC-DCs.
Conclusion Knockdown of CCL5 protein during MDSC-DCs culture might provide a promising method to acquire DC-based tumor vaccines with high antitumor activity.

Keywords

CD24+ MDSC-DCs / CCL5-deficiency / tumor vaccines / antitumor activity / Tregs

Authors' Contributions

Y.Z. and L.H. were involved in conception and design and development of methodology. L.H., ZQ.Z., and Y.Z. helped in acquisition of data (provided animals, acquired and managed patients, provided facilities, etc.). L.H., ZQ.D., W.-Q.G., and Y.Z. contributed substantially in analysis and interpretation of data (e.g., statistical analysis, biostatistics, computational analysis). L.H., ZQ.D., W.-Q.G., and Y.Z. were involved in writing, review, and/or revision of the manuscript. Y.Z. was involved in administrative, technical, or material support. W.-Q. G. and Y.Z. supervised the study.

Funding

This work was supported by funds from the National Natural Science Foundation of China (81773250 and 81972670 to Y.Z.).

Conflict of Interest

None declared.

References

[1] Bremer C.Molecular targets. Encycl Diagnostic Imaging 2008; 1154-1155

[2] van Willigen WW, Bloemendal M, Gerritsen WR, Schreibelt G, de Vries IJM, Bol KF. Dendritic cell cancer therapy: vaccinating the right patient at the right time. Front Immunol 2018; 9(OCT): 2265 DOI: 10.3389/fimmu.2018.02265.

[3] Waldman AD, Fritz JM, Lenardo MJ.A guide to cancer immunotherapy: from T cell basic science to clinical practice. Nat Rev Immunol 2020; 20(11): 651-668

[4] Mildner A, Jung S.Development and function of dendritic cell subsets. Immunity 2014; 40(05): 642-656

[5] Huber A, Dammeijer F, Aerts JGJV, Vroman H.Current state of dendritic cell-based immunotherapy: opportunities for in vitro antigen loading of different DC subsets?. Front Immunol 2018; 9: 2804 DOI: 10.3389/fimmu.2018.02804.

[6] Mastelic-Gavillet B, Balint K, Boudousquie C, Gannon PO, Kandalaft LE.Personalized dendritic cell vaccines-recent breakthroughs and encouraging clinical results. Front Immunol 2019; 10(APR): 766

[7] Kantoff PW, Higano CS, Shore ND.et al; IMPACT Study Investigators. Sipuleucel-T immunotherapy for castration-resistant prostate cancer. N Engl J Med 2010; 363(05): 411-422

[8] Anassi E, Ndefo UA.Sipuleucel-T (provenge) injection: the first immunotherapy agent (vaccine) for hormone-refractory prostate cancer. P&T 2011; 36(04): 197-202

[9] Huber ML, Haynes L, Parker C, Iversen P.Interdisciplinary critique of sipuleucel-T as immunotherapy in castration-resistant prostate cancer. J Natl Cancer Inst 2012; 104(04): 273-279

[10] Higano CS, Schellhammer PF, Small EJ.et al.Integrated data from 2 randomized, double-blind, placebo-controlled, phase 3 trials of active cellular immunotherapy with sipuleucel-T in advanced prostate cancer. Cancer 2009; 115(16): 3670-3679

[11] Sabado RL, Balan S, Bhardwaj N.Dendritic cell-based immunotherapy. Cell Res 2017; 27(01): 74-95

[12] Palucka AK, Ueno H, Connolly J.et al.Dendritic cells loaded with killed allogeneic melanoma cells can induce objective clinical responses and MART-1 specific CD8+ T-cell immunity. J Immunother 2006; 29(05): 545-557

[13] Redman BG, Chang AE, Whitfield J.et al.Phase Ib trial assessing autologous, tumor-pulsed dendritic cells as a vaccine administered with or without IL-2 in patients with metastatic melanoma. J Immunother 2008; 31(06): 591-598

[14] Hinshaw DC, Shevde LA.The tumor microenvironment innately modulates cancer progression. Cancer Res 2019; 79(18): 4557-4566

[15] Nagarsheth N, Wicha MS, Zou W.Chemokines in the cancer microenvironment and their relevance in cancer immunotherapy. Nat Rev Immunol 2017; 17(09): 559-572

[16] Greten FR, Grivennikov SI.Inflammation and cancer: triggers, mechanisms, and consequences. Immunity 2019; 51(01): 27-41

[17] Jordan KR, Kapoor P, Spongberg E.et al.Immunosuppressive myeloid-derived suppressor cells are increased in splenocytes from cancer patients. Cancer Immunol Immunother 2017; 66(04): 503-513

[18] Gabrilovich DI.Myeloid-derived suppressor cells. Cancer Immunol Res 2017; 5(01): 3-8

[19] Grover A, Sanseviero E, Timosenko E, Gabrilovich DI.Myeloid-derived suppressor cells: a propitious road to clinic. Cancer Discov 2021; 11(11): 2693-2706

[20] Aldinucci D, Borghese C, Casagrande N.The ccl5/ccr5 axis in cancer progression. Cancers (Basel) 2020; 12(07): 1-30

[21] Zhang S, Zhong M, Wang C, Xu Y, Gao WQ, Zhang Y.CCL5-deficiency enhances intratumoral infiltration of CD8+ T cells in colorectal cancer. Cell Death Dis 2018; 9(07): 766

[22] Zhang Y, Lv D, Kim HJ.et al.A novel role of hematopoietic CCL5 in promoting triple-negative mammary tumor progression by regulating generation of myeloid-derived suppressor cells. Cell Res 2013; 23(03): 394-408

[23] Younos I, Donkor M, Hoke T.et al.Tumor- and organ-dependent infiltration by myeloid-derived suppressor cells. Int Immunopharmacol 2011; 11(07): 816-826

[24] Bronte V, Brandau S, Chen SH.et al.Recommendations for myeloid-derived suppressor cell nomenclature and characterization standards. Nat Commun 2016; 7: 12150 DOI: 10.1038/ncomms12150.

[25] Perez CR, De Palma M.Engineering dendritic cell vaccines to improve cancer immunotherapy. Nat Commun 2019; 10(01): 5408

[26] Mirza N, Fishman M, Fricke I.et al.All-trans-retinoic acid improves differentiation of myeloid cells and immune response in cancer patients. Cancer Res 2006; 66(18): 9299-9307

[27] Mueller BU, Pabst T, Fos J.et al.ATRA resolves the differentiation block in t(15;17) acute myeloid leukemia by restoring PU.1 expression. Blood 2006; 107(08): 3330-3338

[28] Fang X, Zheng P, Tang J, Liu Y.CD24: from A to Z. Cell Mol Immunol 2010; 7(02): 100-103

[29] Askew D, Harding CV.Antigen processing and CD24 expression determine antigen presentation by splenic CD4+ and CD8+ dendritic cells. Immunology 2008; 123(03): 447-455

[30] Schlitzer A, McGovern N, Teo P. et al. IRF4 transcription factor-dependent CD11b+ dendritic cells in human and mouse control mucosal IL-17 cytokine responses. Immunity 2013; 38(05): 970-983

[31] Kim TS, Gorski SA, Hahn S, Murphy KM, Braciale TJ.Distinct dendritic cell subsets dictate the fate decision between effector and memory CD8(+) T cell differentiation by a CD24-dependent mechanism. Immunity 2014; 40(03): 400-413

[32] Fridman WH, Pagès F, Sautès-Fridman C, Galon J.The immune contexture in human tumours: impact on clinical outcome. Nat Rev Cancer 2012; 12(04): 298-306

[33] Zhang N, Bevan MJ.CD8(+) T cells: foot soldiers of the immune system. Immunity 2011; 35(02): 161-168

[34] Togashi Y, Shitara K, Nishikawa H.Regulatory T cells in cancer immunosuppression - implications for anticancer therapy. Nat Rev Clin Oncol 2019; 16(06): 356-371

[35] Vignali DAA, Collison LW, Workman CJ.How regulatory T cells work. Nat Rev Immunol 2008; 8(07): 523-532

[36] Chen ML, Pittet MJ, Gorelik L.et al.Regulatory T cells suppress tumor-specific CD8 T cell cytotoxicity through TGF-β signals in vivo. Proc Natl Acad Sci U S A 2005; 102(02): 419-424

[37] Setiady YY, Coccia JA, Park PU.In vivo depletion of CD4+FOXP3+ Treg cells by the PC61 anti-CD25 monoclonal antibody is mediated by FcgammaRIII+ phagocytes. Eur J Immunol 2010; 40(03): 780-786

[38] Zhang Q, Qin J, Zhong L.et al.CCL5-mediated Th2 immune polarization promotes metastasis in luminal breast cancer. Cancer Res 2015; 75(20): 4312-4321

[39] Altevogt P, Sammar M, Hüser L, Kristiansen G.Novel insights into the function of CD24: a driving force in cancer. Int J Cancer 2021; 148(03): 546-559

[40] Barkal AA, Brewer RE, Markovic M.et al.CD24 signalling through macrophage Siglec-10 is a target for cancer immunotherapy. Nature 2019; 572(7769): 392-396

RIGHTS & PERMISSIONS

Global Medical Genetics