4A and Table 1). Spleen and
lymph nodes of IgM or JH KO rats showed barely detectable IgM or IgD positive cells (Fig. 4A, Table 1 and Supporting Information Data 4). The total number of cells in the spleen and lymph nodes of IgM or JH KO rats were drastically decreased versus WT rats (Table 1). IgM+ and CD45R+cells in the spleen of IgM or JH KO rats were drastically decreased versus WT rats (IgM+: 0.7 and 2.28%, respectively; CD45R+: 1.6 and 4.3%, respectively) (Table 1). FACS analysis showed the presence of a small population of CD45R+IgM− cells in spleen (Fig. 4A, Table 1). Immunohistology revealed their location mainly in the spleen red pulps IWR-1 (data not shown). Using several markers, we confirmed that the phenotype of CD45R+ cells in IgM KO rats corresponded to the previously described phenoype of rat pDC 18 (data not shown). In lymph nodes, absolute numbers
of IgM+ or CD45R+ cells were greatly reduced in IgM or JH KO rats versus WT controls (∼4 and ∼4.5%, respectively) (Table 1). In BM of IgM or JH KO rats, we observed no immature or mature B cells and greatly reduced proportion of pro–pre B cells Ivacaftor datasheet (IgM− CD45Rlow) (Fig. 4A). The absolute number of mononuclear cells was significantly reduced in IgM and JH KO versus WT rats (42.2 and 56.7%, respectively) (Table 1) and numbers of pro–pre B cells (IgM− CD45Rlow) in IgM, JH KO and WT were 12.8 and 22.4%, respectively, versus WT (Table 1). T cells in spleen, as defined by double staining using anti-TCRαβ and anti-CD4 or anti-CD8 Ab, showed an increased proportion Meloxicam of TCRαβ+ cells compared with WT rats (∼85% in IgM and JH KO rats versus ∼40% in WT animals), both of the CD4+ and CD8+ subtypes (Fig. 4B). Despite this increase, the total numbers of spleen cells in IgM and JH KO rats were only 13.6 and 16.6%, respectively, compared with WT spleen cells and thus the total numbers of TCRαβ+ cells in IgM and JH KO rats were 30 and 33.7%, respectively, versus WT (p=<0.05 for both IgM or JH KO versus WT) (Table 1). Despite the fact that cell numbers in the lymph nodes were considerably decreased in IgM or JH KO versus WT rats (43
and 39%, respectively), T cells were not significantly reduced (Table 1) due to a significantly increased proportion of TCRαβ+ cells (∼95% for both KO versus ∼78%, respectively) with the CD4+ or CD8+ surface marker (Supporting Information Data 2). In BM, the proportion of TCR+ cells was increased in IgM or JH KO versus WT rats (both ∼35 versus ∼10%, respectively) in both compartments, TCR+CD4+ and TCR+CD8+ (Supporting Information Data 2). The total number of T cells was also significantly increased in IgM or JH KO versus WT (275 and 201%, respectively) (Table 1). In thymus of IgM or J KO rats, the proportion of TCR+, TCR+CD4+ and TCR+CD8+ cells (Supporting Information Data 3) as well as the total number of T cells (Table 1) were comparable.