Abstract The precise mechanisms that govern the commitment of CD4 T cells to become Th1 or Th2 cells in vivo are incompletely understood. Recent experiments demonstrate colocalization of the IFN-γR chains with the TCR during activation of naive CD4 T cells, suggesting that association of these molecules may be involved in determining lineage commitment. To test the role of IFN-γ and its receptor in the generation of Th1 Ag-specific CD4 T cells, we analyzed mice after infection with Listeria monocytogenes or lymphocytic choriomeningitis virus. In the absence of IFN-γ, Ag-specific CD4 T cells were generated in response to both these infections. In addition, IFN-γ-producing (Th1) Ag-specific CD4 T cells were generated in mice lacking the ligand-binding chain of the IFN-γR (IFN-γR1−/−) or the signaling chain (IFN-γR2−/−). There was no increase in the number of IL-4-producing Ag-specific CD4 T cells, nor was there a decrease in the expression of T-bet in the absence of functional IFN-γ signaling, indicating that the cells were committed Th1 cells. Thus, both chains of the IFN-γR are dispensable for the generation of Th1 Ag-specific CD4 T cells after infection in vivo.
Proinflammatory cytokines, such as gamma interferon (IFN-gamma), impact aspects of T-cell responses after infection, including expansion, contraction, and memory formation. Interleukin-18 (IL-18) functions as a proinflammatory cytokine by stimulating the production of IFN-gamma from multiple cell types and accentuating the development of Th1 CD4 T-cell responses. Focused microarray analyses revealed upregulation of IL-18 and IL-18 receptor genes in CD8 T cells during the contraction phase. Based on these findings we investigated if and how signaling through the IL-18 receptor influences the development and kinetics of antigen (Ag)-specific CD8 and CD4 T-cell responses following infection. IL-18Ralpha(-/-) and IL-18(-/-) mice developed frequencies and total numbers of Ag-specific CD8 T cells after Listeria monocytogenes infection that were similar to those of wild-type C57BL/6 mice. The kinetics of expansion, contraction, and memory CD8 T-cell maintenance were also similar. When IL-18Ralpha deficiency was isolated to Ag-specific CD8 T cells, the kinetics of the expansion and contraction phases were also normal. These basic findings were confirmed by examining the response to vaccinia virus infection. In contrast, the expansion of Ag-specific CD4 T cells was slightly curtailed by the absence of IL-18Ralpha; however, contraction and the maintenance of memory were not altered. Importantly, both memory Ag-specific CD8 and CD4 T cells generated in the absence of IL-18Ralpha expanded appropriately after secondary antigen exposure and were protective, indicating that signaling through the IL-18 receptor is not required for normal T-cell response kinetics and survival of immunized mice challenged with a lethal L. monocytogenes infection.
Abstract This study investigates vasoactive intestinal peptide receptor 1 (VPAC1) and VPAC2 regulation during the immune response of murine T cells. VPAC1 is an anti-proliferative, G-protein coupled receptor that has been shown to be downregulated upon ex vivo T cell activation, whereas VPAC2 was shown to be upregulated. To date, regulation of VPAC1 and 2 have not been elucidated in primary, murine T cells throughout the phases of a T cell immune response following infection and whether they have a role in memory T cell evolution. Splenic CD4 T cells were isolated from C57Bl/6J mice 24hr post in vivo activation with anti-CD3. Flow cytometry (Accuri) analysis confirmed 90% activation of cells (CD69+/CD25+). QPCR analysis of total RNA showed VPAC1 levels decreased 93% in activated CD4 T cells, while VPAC2 levels increased 66%, possibly suggesting that these receptors regulate early components of the CD4 T cell response. These results prompted us to pursue an in vivo activation model utilizing OT-I and OT-II T cells. We hypothesize in CD4 T cells, VPAC1 levels will decrease upon activation and gradually increase to naïve levels throughout contraction into memory phase, while VPAC2 levels will increase upon activation and gradually decrease to lower naïve levels. The reciprocal expression pattern of these receptors may indicate they have opposing functions in regulating the kinetics of the T cell response and memory cell evolution. 1KO1 DK064828, COBRE- 2P20RR015566
Mice infected with the murine coronavirus, mouse hepatitis virus, strain JHM (MHV) develop an immune-mediated demyelinating encephalomyelitis. Adoptive transfer of MHV-immune splenocytes depleted of either CD4 or CD8 T cells to infected mice deficient in recombination activation gene 1 resulted in demyelination. We showed previously that the process of CD8 T-cell-mediated demyelination was strongly dependent on the expression of gamma interferon (IFN-gamma) by donor cells. In this report, we show, in contrast, that demyelination and lymphocyte infiltration were increased in recipients of IFN-gamma(-/-) CD4 T cells when compared to levels in mice receiving C57BL/6 CD4 T cells.
