B2 (2018 - 2021) - C5a Receptor 2 Regulates NK Cell Function in Pregnancy

In experimental sepsis, C5ar1- or NK cell-deficient mice showed increased survival associated with decreased TNF-alpha and IFN-gamma production. The decreased cytokine production results from C5aR-mediated regulation of DC and NK cell functions, suggesting that C5a faciliates the deleterious immune responses in sepsis by modulating NK/DC cross-talk. Further, C5a is critical for the development of appropriate CD4+ T cell responses in L.major and T.gondii infection as well as CD8+ T cell responses in influenza or LCMV infection. However, the molecular mechanisms underlying C5aR-mediated regulation of NK cell receptor functions in infection and its impact for the induction of antigen-specific CD4+ and CD8+ T cell responses remain elusive. Data from Fusakio et al. suggest that C5a regulates the expression of NKp46. Using N-Ethyl-N-nitrosourea (ENU), two germline mutants, designated Ace and Chip, have been identified in the Hoebe lab. Coarse mapping and next generation sequencing revealed the Chip phenotype to result from a missense mutation in the Immunoreceptor tyrosine-based switch motif (ITSM) domain of the signaling lymphocyte activation molecule (SLAM)-related receptor 2B4 (CD244) that can elicit both activating as well as inhibitory signals on NK cells. The causal variant in the Ace mutant was found to be a missense mutation in the SH2 domain of Slp76, an adapter molecule for which the function in NK cells is poorly defined. Interestingly, Ace mutants showed no abnormalities in T cell development or T cells responses. Both strains have normal NK cell development but are susceptible to T.gondii infection as a result of increased or decreased NK cell-dependent immune responses. Here, we aim at investigating the role of C5aR signalling in NK cell function and its interaction with specific NK cell receptors identified through ENU mutagenesis in experimental T. gondii infection.


1. Analyze expression patterns of C5aR2 and NKp46 in pregnant wt, C5aR2-deficient and tdTomato-C5aR2 reporter mice at different pregnancy stages (e.g. gestation day 5, 7, 10, 12) under steady state conditions with a specific focus on, but not limited to uNK cells

2. Compare different factors contributing to the success of pregnancy (distribution of uNK cells, impact on spiral arteries and production of cytokines) in wt and C5aR2-deficient mice at different gestation days (see Aim 1) to elucidate exact time point and stage of pregnancy failure

3. Analyze the impact of aToxoplasma gondii infection on the expressions patterns and function of C5aR2 and NKp46 in uNK cells in pregnant wt and C5aR2-deficient mice