The intrinsic reasons between two models might be quite similar. 104 purified CD4+ T cells from PBS or pristane-treated BALB/c mice were cultured with 9 104 CD4? splenocytes from wild-type BALB/c mice or the splenocytes from TCRcoculture for 8 days. 2.7. Statistical Analysis Data were presented as means standard?error of means (S.E.M). Statistical analyses were performed by using GraphPad Prism 5.0 software (GraphPad Prism, San Diego, CA, USA). Statistic difference was determined by the unpaired Student < 0.05 was considered statistically significant. 3. Results 3.1. Hyperglobulinemia in Pristane-Induced Lupus-Like Mice Increase in serum autoAbs levels is one of the key manifestations of SLE. One dose injection of pristane largely recapitulates the pathology of SLE in BALB/c mice. Serum Ig, including total IgM, IgG, IgG1, and IgG2a levels in pristane-induced mice, were detected at 6 months by an ELISA assay. It was shown that total IgM and IgG levels of pristane-treated mice were higher than those in PBS-treated control mice at 6 months (Figures 1(a) and 1(b)). IgG2a autoAb is considered to be Elacestrant the most pathogenic, whereas IgG1 displays the poorest pathogenicity in the mice [26]. In our study, we found that IgG2a level in pristane-treated mice increased significantly Elacestrant at 6 months after the injection (Physique 1(c)). More specifically, autoAbs to P0, dsDNA, and SnRNP were elevated in the sera of pristane-treated mice when compared to control mice (Figures 1(d)C1(f)) with more deposition of ICs in the kidneys consequently (Figures 1(g) and 1(h)). These results confirm the presence of hyperglobulinemia in pristane-treated mice, which is associated with lupus-like pathogenesis. Rabbit Polyclonal to p73 Open in a separate window Physique 1 Serum immunoglobulin levels in pristane-induced lupus-like mice. Sera were collected before the injection and at 6 months post-PBS or pristane treatment. The levels of total IgG (a), total IgM (b), IgG2a (c), and anti-P0 (d), anti-SnRNP (e), and anti-dsDNA (f) autoantibodies in the serum were determined by an ELISA assay. Furthermore, the kidneys were collected when the mice were sacrificed at 6 months after PBS/pristane injection. The deposition of immune complexes (g) in the kidney was detected by the immunofluorescence assay. (h) Statistic analysis of immune complexes in the kidneys. Data were pooled from three impartial experiments with 5 mice per group in each experiment. ?< 0.05; ??< 0.01; ???< 0.001; ????< 0.0001. 3.2. Increased Expression of Activated Markers on CD4+ T Cells and B Cells in the Spleens of Pristane-Induced Lupus-Like Mice Antibody-producing cells (also called as plasma cells) can arise through either direct differentiation in EF or GCs with the help of CD4+ T cell [27]. There exist increased numbers of circulating pre-GC B cells, CCR7loPD-1hiCXCR5+CD4+ T cells, and memory B cells in SLE patients as well as in lupus-like mice, indicating the presence of CD4+ T cell activation and subsequently enhanced GC responses [28C30]. This is associated with elevated generation of plasma cells within GCs and contributes to the development of lupus pathogenesis in both mice and humans [27]. Therefore, we tested the phenotypes of CD4+ T cells and B cells derived from pristane and PBS-treated mice by flow cytometry. It was apparent that this expression of CD44 increased significantly whereas that of CD62L decreased significantly on CD4+ Elacestrant T cells from the pristane-treated group when compared to the PBS group (Figures 2(a) and 2(b)). More interestingly, costimulatory molecules, which are important for T-B conversation including ICOS, OX40, and PD-1, were also elevated on CD4+ T cells from pristane-treated mice (Figures 2(a) and 2(b)). These results indicate that CD4+ T cells derived from pristane-treated mice display activated phenotypes, which might facilitate B cell activation and differentiation. Open in a separate window Physique 2 Activated phenotypes of CD4+ T cells in the pristane-induced lupus-like mouse model. The splenocytes were collected at 6 months after i.p. injection of PBS or pristane. (a) The expression levels of CD62L, CD44, ICOS, OX40, and PD-1 on CD4+ T cells. (b) Statistic percentages of.