Regulation of arginase I activity and expression by both PD-1 and CTLA-4 on the myeloid-derived suppressor cells - PubMed (original) (raw)
Regulation of arginase I activity and expression by both PD-1 and CTLA-4 on the myeloid-derived suppressor cells
Yu Liu et al. Cancer Immunol Immunother. 2009 May.
Abstract
An elevated number of Gr-1(+)CD11b(+) myeloid-derived suppression cells (MDSCs) has been described in mice and human bearing tumor and associated with immune suppression. Arginase I production by MDSCs in the tumor environment may be a central mechanism for immunosuppression and tumor evasion. In this study and before, we found that Gr-1(+)CD11b(+) MDSCs from ascites and spleen of mice bearing ovarian 18D carcinoma express a high level of PD-1, CTLA-4, B7-H1 and CD80 while other co-stimulatory molecules, namely CD40, B7-DC and CD86 are not detected. Further studies showed that PD-1 and CTLA-4 on the Gr-1(+)CD11b(+) MDSCs regulated the activity and expression of arginase I. The blockage and silencing of PD-1, CTLA-4 or both PD-1 and CTLA4 molecules could significantly reduce arginase I activity and expression induced with tumor-associated factor. Similar results were also observed while their ligands B7-H1 and/or CD80 were blocked or silenced. Furthermore, CD80 deficiency also decreased the arginase I expression and activity. Antibody blockade or silencing of PD-1, CTLA-4 or both reduced the suppressive potential of PD-1+CTLA-4+MDSCs. Blockade of PD-1, CTLA-4 or both also slowed tumor growth and improved the survival rate of tumor-bearing mice. Thus, there may exist a coinhibitory and costimulatory molecules-based immuno-regulating net among MDSCs.
Figures
Fig. 1
PD1+CTLA-4+MDSCs derived from ascites of mice bearing ovarian 1D8 carcinoma express arginase I. a Expression of PD1 and CTLA-4 on the surface of murine carcinoma-associated Gr1+MDSCs cells. A1, A2 MDSCs from ascites of mice bearing ovarian 1D8 carcinoma. A3, A4 Gr1+cells from naive mouse splenic cells. b Isolated PD1+CTLA-4+Gr1+CD11b+MDSCs from ascites of mice bearing ovarian carcinoma (Giemsa staining). PD-1+CTLA-4+MDSCs were sorted from the ascites of mice with ovarian 1D8 carcinoma using PE-labeled anti-Gr1 and FITC-labeled anti-CD11b using flow cytometry and the sorted cells were stained using Giemsa stain kit (Thermo Fisher Scientific, USA) according to the manufacture’s procedure. c Murine ovarian carcinoma supernatants promoted the arginase I activity and expression by PD-1+CTLA-4+MDSCs. Arginase activity (C1) and transcription level (C2) of arginase I in PD-1+CTLA-4+MDSCs upon exposing to different concentration of murine ovarian carcinoma supernatant and mrIL-13 (positive control). Cell contr. CD11b+Gr1+ cells from naive mouse splenic cells, MDSCs PD1+CTLA-4+MDSCs from the ascites of mice bearing ovarian carcinoma, Tu. Sup. tumor supernatants from ovarian 1D8 carcinoma after culturing for 24 h
Fig. 2
Blockade of PD-1, CTLA-4 or their ligands decreases arginase I activity and expression by ovarian carcinoma-associated MDSCs. a Blockade of PD-1, CTLA-4 or both reduced arginase I activity and expression. Anti-PD-1, CTLA-4 or both antibodies were, respectively, added into the culture of PD-1+CTLA4+MDSCs in the conditioned medium and then their arginase I activity and transcription level were analyzed after 24 h. b Blockade of B7-H1 and CD80 decreased the arginase I activity and transcription. Similarly, Anti-CD80, B7-H1 or both antibodies were added into the culture of PD-1+CTLA4+MDSCs in the conditioned medium and then their arginase I activity and transcription level were analyzed after 24 h. The isolated PD-1+CTLA4+MDSCs were plated at 1 × 106 cells/well in 24-well tissue culture plates and stimulated with equal amount of tumor supernatants (25% V/V). Following stimulation, cells were washed with PBS and lysed in lysis buffer described in “Materials and methods”. Tu. Sup. Tumor supernatants from ovarian 1D8 carcinoma after culturing for 24 h
Fig. 3
Knockdown of PD-1, CTLA-4 or both decreases arginase I activity and expression. a PD-1 and CTLA4 siRNA reduced the level of PD-1 and CTLA-4, respectively, but not that of GAPDH transcript expression in the transfected ovarian carcinoma-associated PD-1+CTLA-4+MDSCs, as assessed by RT-PCR. b PD-1, CTLA4 or both siRNA but not non-specific siRNA, reduced the level of surface PD-1 and CTLA4 expression in transfected ovarian carcinoma-associated PD-1+CTLA-4+MDSCs, as assessed by flow cytometric analysis, respectively. B1, B2 Expression of PD-1 and CTLA-4 on the siRNA transfected ovarian carcinoma-associated PD-1+CTLA-4+MDSCs, respectively. Black line Isotype control, dark black line expression of PD-1 (B1) and CTLA-4 (B2) on ovarian carcinoma-associated PD-1+CTLA-4+MDSCs transfected with PD-1 and CTLA-4 specific siRNA, respectively, dotted line expression of PD-1 (B1) and CTLA-4 (B2) on ovarian carcinoma-associated PD-1+CTLA-4+MDSCs transfected by control mock siRNA. c Knockdown of PD-1, CTLA-4 or both reduced arginase I activity (C1) and level of transcription (C2). PD-1, CTLA-4 or both siRNA transfected PD1+CTLA-4+ MDSCs were cultured in the conditioned medium for 24 h and then lysed for analysis of arginase I activity. The total RNA was extracted for transcription assay according to the described protocol in “Materials and methods”. T. Sup. Conditioned medium containing 25% murine ovarian carcinoma supernatant (V/V), PD1siRNA, CTLA4siRNA and PD1+CTLA4siRNA, respectively, were PD1 siRNA, CTLA4 siRNA or both transfected PD1+CTLA4+MDSCs, Mock siRNA control siRNA transfected PD1+CTLA4+MDSCs
Fig. 4
CD80 deficiency decreases arginase I activity and expression by ovarian carcinoma-associated PD1+CTLA-4+ MDSCs. a The activity and expression of fresh CD11b+Gr1+ cells from wild type (WT) or CD80 deficiency (CD80−/−) spleen of mouse with (Tumor) or without ovarian carcinoma (Contr.). Activity (A1) and transcriptional level (A2) of arginase I were analyzed in WT mice or CD80−/− mice by the amount of urea released from cell extract and RT-PCR. b CD80−/− reduced the arginase activity (B1) and transcription (B2) induced using tumor supernatant and mrIL13. Contr. PD1+CTLA-4+MDSCs cells alone, Tu.Sup. PD1+CTLA-4+ MDSCs were cultured in conditioned medium containing 25% tumor supernatant, and mrIL13 PD1+CTLA-4+ MDSCs were cultured in 10 ng/ml IL13 medium
Fig. 5
Antibody blockade or siRNA silencing of PD-1 and CTLA-4 reduces the suppressive potential of PD-1+CTLA-4+ MDSCs. a Effect of pretreatment of PD-1+CTLA-4+ MDSCs using neutralizing antibody PD-1, CTLA-4 or both on the generation of IFNγ by VLP-specific splenocytes (VLP.SC). Supernatants of cultures were collected after 24 h and IFNγ was assayed by capture ELISA. Isotypic contr. Control antibody. b PD-1, CTLA-4 or both siRNA transfected MDSCs had the reduced suppression on VLP-induced generation of IFN-γ in VLP-specific splenocyte culture. PD-1+CTLA-4+ MDSCs were sorted from the ascites of mice with ovarian 1D8 carcinoma and then transfected using PD-1, CTLA-4 or both siRNA or mock siRNA and then added into VLP-specific splenocyte culture. PD-1+, CTLA-4+ or CTLA-4 and PD1 siRNA represented PD-1+CTLA-4+MDSCs, which were transfected with PD-1, CTLA-4 or both siRNA, respectively, mock siRNA PD-1+CTLA-4+ MDSCs transfected with control siRNA
Fig. 6
Blockade of PD-1 or CTLA-4 slows tumor growth and improves the survival rate of mice bearing tumor. a In vivo injection of neutralizing antibody to PD-1, CTLA-4 or both, significantly slowed and retarded tumor growth. b In vivo injection of neutralizing antibody to PD-1, CTLA-4 or both, significantly prolonged the survival rate. Mice (6/group) were injected using 1D8 cells (s.c. with 1 × 107) and i.p. with 250 μg of control antibody or antibody specific for PD-1 and CTLA-4 or both on days −6, −4, −2 and +1 as previously described [20]
Fig. 7
Schematic illustration of a mechanism by which PD-1 and B7-H1 on MDSCs subvert antitumor immunity via regulating the activity and expression of arginase I. MDSCs were induced by tumor-derived factors such as VEGF, TGF-β, IL-6 and PEG2 etc. These tumor-associated MDSCs express higher level of PD-1, CTLA4 and also B7-H1 and CD80, which are ligands of PD-1 and CTLA-4, respectively. Both PD-1 and CTLA4 on the surface of MDSCs may upregulate the activity and expression of arginase I via binding with their ligands expressed on the MDSCs. These upregulated arginase I can inhibit the activation of CD4+ and CD8+ T cells to regulate immune responses. BMC Bone marrow cells
Similar articles
- B7-H1 on myeloid-derived suppressor cells in immune suppression by a mouse model of ovarian cancer.
Liu Y, Zeng B, Zhang Z, Zhang Y, Yang R. Liu Y, et al. Clin Immunol. 2008 Dec;129(3):471-81. doi: 10.1016/j.clim.2008.07.030. Epub 2008 Sep 14. Clin Immunol. 2008. PMID: 18790673 - Analysis of the role of negative T cell costimulatory pathways in CD4 and CD8 T cell-mediated alloimmune responses in vivo.
Ito T, Ueno T, Clarkson MR, Yuan X, Jurewicz MM, Yagita H, Azuma M, Sharpe AH, Auchincloss H Jr, Sayegh MH, Najafian N. Ito T, et al. J Immunol. 2005 Jun 1;174(11):6648-56. doi: 10.4049/jimmunol.174.11.6648. J Immunol. 2005. PMID: 15905503 - LAG-3, TGF-β, and cell-intrinsic PD-1 inhibitory pathways contribute to CD8 but not CD4 T-cell tolerance induced by allogeneic BMT with anti-CD40L.
Lucas CL, Workman CJ, Beyaz S, LoCascio S, Zhao G, Vignali DA, Sykes M. Lucas CL, et al. Blood. 2011 May 19;117(20):5532-40. doi: 10.1182/blood-2010-11-318675. Epub 2011 Mar 21. Blood. 2011. PMID: 21422469 Free PMC article. - Interaction of PD-L1 on tumor cells with PD-1 on tumor-specific T cells as a mechanism of immune evasion: implications for tumor immunotherapy.
Blank C, Gajewski TF, Mackensen A. Blank C, et al. Cancer Immunol Immunother. 2005 Apr;54(4):307-14. doi: 10.1007/s00262-004-0593-x. Epub 2004 Dec 15. Cancer Immunol Immunother. 2005. PMID: 15599732 Free PMC article. Review. - Targeting T cell costimulation in autoimmune disease.
Stuart RW, Racke MK. Stuart RW, et al. Expert Opin Ther Targets. 2002 Jun;6(3):275-89. doi: 10.1517/14728222.6.3.275. Expert Opin Ther Targets. 2002. PMID: 12223069 Review.
Cited by
- Identification of Specific Cell Surface Markers on Immune Cells of Squirrel Monkeys (Saimiri sciureus).
Nehete BP, DeLise A, Nehete PN. Nehete BP, et al. J Immunol Res. 2024 Mar 25;2024:8215195. doi: 10.1155/2024/8215195. eCollection 2024. J Immunol Res. 2024. PMID: 38566886 Free PMC article. - Myeloid-derived suppressor cells promote allograft survival by suppressing regulatory T cell dysfunction in high-risk corneal transplantation.
Lee S, Blanco T, Musayeva A, Dehghani S, Narimatsu A, Forouzanfar K, Ortiz G, Kahale F, Wang S, Chen Y, Dohlman TH, Chauhan SK, Dana R. Lee S, et al. Am J Transplant. 2024 Sep;24(9):1597-1609. doi: 10.1016/j.ajt.2024.03.022. Epub 2024 Mar 19. Am J Transplant. 2024. PMID: 38514014 - A Vaccine against Cancer: Can There Be a Possible Strategy to Face the Challenge? Possible Targets and Paradoxical Effects.
Zefferino R, Conese M. Zefferino R, et al. Vaccines (Basel). 2023 Nov 8;11(11):1701. doi: 10.3390/vaccines11111701. Vaccines (Basel). 2023. PMID: 38006033 Free PMC article. Review. - Metabolic reprogramming of the tumor immune microenvironment in ovarian cancer: A novel orientation for immunotherapy.
Lin Y, Zhou X, Ni Y, Zhao X, Liang X. Lin Y, et al. Front Immunol. 2022 Oct 14;13:1030831. doi: 10.3389/fimmu.2022.1030831. eCollection 2022. Front Immunol. 2022. PMID: 36311734 Free PMC article. Review. - Opportunities for Nitric Oxide in Potentiating Cancer Immunotherapy.
Kim J, Thomas SN. Kim J, et al. Pharmacol Rev. 2022 Oct;74(4):1146-1175. doi: 10.1124/pharmrev.121.000500. Pharmacol Rev. 2022. PMID: 36180108 Free PMC article. Review.
References
- Young MR, Wright MA, Matthews JP, Malik I, Prechel M. Suppression of T cell proliferation by tumor-induced granulocyte-macrophage progenitor cells producing transforming growth factor-beta and nitric oxide. J Immunol. 1996;156:1916–1922. - PubMed
- Kono K, Salazar-Onfray F, Petersson M, Hansson J, Masucci G, Wasserman K, Nakazawa T, Anderson P, Kiessling R. Hydrogen peroxide secreted by tumor-derived macrophages down-modulates signal-transducing zeta molecules and inhibits tumor-specific T cell-and natural killer cell-mediated cytotoxicity. Eur J Immunol. 1996;26:1308–1313. doi: 10.1002/eji.1830260620. - DOI - PubMed
- Fu YX, Watson GA, Kasahara M, Lopez DM. The role of tumor-derived cytokines on the immune system of mice bearing a mammary adenocarcinoma. I. Induction of regulatory macrophages in normal mice by the in vivo administration of rGM-CSF. J Immunol. 1991;146:783–789. - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Medical
Research Materials