Inhibition of NLRC5 regulates cytokine expression in CD4+ T helper lymphocytes and prolongs murine islet and skin allograft survival

Mol Immunol. 2021 Sep;137:67-74. doi: 10.1016/j.molimm.2021.06.020. Epub 2021 Jul 2.

Abstract

The purpose of this study was to study the effect of inhibiting NLRC5 expression and function on CD4 + T cells, and islet and skin transplantation in mice. A murine skin graft model and islet cell transplantation model were established, and the expression of NLRC5 was compared in rejection and immune tolerance groups. Mice spleen-derived CD4 + T cells were cultured, purified, and enriched in vitro, and transfected with the shRNA lentiviral vector NLRC5-RNAi-GFP. Changes in cytokine secretion were detected to understand changes in immunological function. Murine islet and skin transplantation models were injected with CD4 + T cells transfected with the lentivirus, and the survival time of the grafts and the levels of IFN-γ and IL-10 were compared between groups. The expression of NLRC5 mRNA in islet and skin grafts was significantly increased. In vitro experiments showed that the expression of IL-4 and IL-10 was up-regulated in CD4 + T cells, and T cells differentiation turned to Th2 after inhibition of NLRC5. In vivo experiments showed that inhibition of NLRC5 prolonged islet and skin graft survival. Pathological examination showed that the rejection of transplanted skin and islets in the NLRC5-RNAi group was mild, and there was a correlation between high expression of NLRC5 and rejection of mouse islet and skin grafts. In summary, inhibition of NLRC5 can prolong islet and skin graft survival induce transplant immune tolerance through induction of the secretion of Th2 cytokines by CD4 + T cells.

Keywords: CD4+T cells; Islet transplantation; Lentivirus; NLRC5; Skin transplantation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allografts / immunology*
  • Animals
  • CD4-Positive T-Lymphocytes / immunology*
  • Cytokines / immunology*
  • Graft Rejection / immunology
  • Graft Survival / immunology*
  • Immune Tolerance / immunology
  • Interferon-gamma / immunology
  • Interleukin-10 / immunology
  • Intracellular Signaling Peptides and Proteins / immunology*
  • Islets of Langerhans Transplantation / methods
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • RNA, Messenger / immunology
  • Skin Transplantation / methods
  • T-Lymphocytes, Helper-Inducer / immunology*
  • Th2 Cells / immunology
  • Transplantation Tolerance / immunology
  • Transplantation, Homologous / methods

Substances

  • Cytokines
  • Intracellular Signaling Peptides and Proteins
  • NLRC5 protein, mouse
  • RNA, Messenger
  • Interleukin-10
  • Interferon-gamma