We have generated LAIR-1 -/- mice on a B6 background to study the in vivo function of LAIR-1. Phenotypic analysis of lymphoid organs did not show large differences between LAIR-1 +/+ and LAIR-1 -/- animals. We have observed a slight increase in the percentage of splenic B cells in the LAIR-1 -/- mice, along with a decrease in T cells, mostly because of a decrease in CD8 T cells. This probably does not result from abnormal trafficking of T lymphocytes, since LAIR-1 +/+ and LAIR-1 -/- T cells traffic equally to peripheral lymphoid organs. In the gut we have observed a slight increase of T cells in the LAIR-1 -/- mice, along with an increase in the NKG2D expression. In vitro experiments showed that OT-II LAIR-1-/- CD4 T cells proliferated less than the OT-II LAIR-1+/+ CD4 T cells when they are cultured with APC loaded with OT-II OVA peptide. To study the immune response in vivo, we have immunized the animals with TNP-OVA and found that class switching is affected in LAIR-1 -/- mice. These animals produced lower levels of IgG2a and IgG2b, while switching to IgG1 is not affected. By using T cell specific LAIR-1 -/- animals (CD4 Cre LAIR-1flox/flox), we confirmed that the defect in class switching is T cell specific. Previous investigators have published that mouse B cells do not express LAIR-1; however, recently we have found that marginal zone B cells are positive for LAIR-1 expression. Other preliminary results have shown that in the LAIR-1 -/- mice there are significant alterations in the recruitment of macrophages and eosinophils into the peritoneum in a model of chemical peritonitis, suggesting a role for LAIR-1 in the trafficking of these cell types towards sites of inflammation.? ? We showed that human neutrophils from peripheral blood express CD300a. To study expression during neutrophil development, we used the HL-60 differentiation model. We showed that CD300a expression is acquired during development and that cell surface expression increases very rapidly when neutrophils are stimulated with LPS and GM-CSF. This increase is the result of translocation of an intracellular pool of the receptor to the cell surface. This ready availability of CD300a is reminiscent of CTLA-4 and suggests that CD300a could play an important role in modulating neutrophil responses. Co-ligation of CD300a with the ITAM containing CD32a (FcRIIa) activation receptor inhibited CD32a mediated signaling, whereas it did not inhibit toll-like receptor (TLR)-4 mediated reactive oxygen species (ROS) production. Therefore, at least for human neutrophils, it seems that the inhibitory signals mediated by the CD300a receptor may be selective in their action.
