Balb/c小鼠脾脏B10细胞分析

目的分析年龄、性别和环境因素对Balb/c小鼠脾脏B10细胞的影响。方法以年龄、性别和不同环境对小鼠进行分组,流式细胞仪分析小鼠脾脏CD5+CD1dhiB细胞的比例。体外培养小鼠脾细胞,在不加任何刺激或LPS+PIM刺激5 h后,流式细胞仪分析CD19+IL-10+B细胞比例。结果 6月龄小鼠脾脏中CD5+CD1dhiB细胞比例平均为2.9%,明显高于2月龄小鼠的2.28%(P<0.05);在不加任何刺激的情况下,两组小鼠脾脏中IL-10+B细胞的比例均很低且无统计学差异(P>0.05);加入LPS+PIM刺激5 h后,6月龄小鼠脾脏中IL-10+B细胞的比例平均为1.93%,高于2月龄小鼠的1.25%(P<0.05)。细胞表型和细胞内IL-10染色分析均显示年龄匹配的雌性和雄性小鼠脾脏B10细胞的表达无明显差异(P>0.05)。普通级和SPF级小鼠CD5+CD1dhiB细胞和LPS+PIM诱导的IL-10+B细胞比例无明显差异(P>0.05),而在不加任何刺激的情况下,普通级小鼠脾脏IL-10+B细胞比例平均为0.87%,明显高于SPF小鼠的0.38%(P<0.01)。结论脾脏B10细胞可随Balb/c小鼠年龄增大而扩增,B10细胞的表达与性别差异无明显关系,微生物环境可促进B10细胞的活化,但对B10细胞总数无明显影响。

Analysis of the splenic B10 cells of Balb/c mice

B10 cells,a type of recently identified regulatory B cells with a phenotype as CD5+CD1dhiCD19hi,regulate T cell mediated autoimmunity and inflammation in an IL-10-dependent manner.However,the development and the activation characteristics of B10 cells remain unclear.The aim of the present study is to explore the influence of the gender,age or environment on the spleenic B10 cells of Balb/c mice.The mice were divided into different groups according to gender,age or different environment,then analyzed the percentage of the CD5+CD1dhi B cells with flow cytometry.Spleen cells of the Balb/c mice were cultured in vitro without stimulation or with LPS+PIM stimulation for 5 h,then analyzed the percentages of the CD19+IL-10+ B cells among total splenic B cells with flow cytometry.We found that the percentage of splenic CD5+CD1dhi B cells of 6-month-old mice(2.9%) was significantly higher than that of 2-month-old mice(2.28%)(P < 0.05).Without stimulation,the frequency of CD19+IL-10+ demonstrated no difference between 2-month-old and 6-month-old mice(P>0.05).After LPS+PIM stimulation for 5 h,the percentages of CD19+IL-10+ B cells of 2-month-old and 6-month-old mice were respectively raised to 1.25% and 1.93%,and the percentage of CD19+IL-10+ B cells of 6-mo-old mice was higher than that of 2-month-old mice(P > 0.05).There was no significantly difference about B10 cell frequency between male and female mice by analyzing with cell phenotype and intra-cellular IL-10 staining(P > 0.05).The percentages of CD5+CD1dhi B cells and IL-10+ B inducing by LPS+PIM stimulation for 5 h were not significantly difference between conventional mice and SPF mice,however the percentage of splenic IL-10+ B cells among total B cells of conventional mice(0.87%) was significantly higher than that of SPF mice(0.38%)(P < 0.01).This data indicates that the splenic B10 cells expand with the increase of age in Balb/c mice,while the gender has no effect on the expression of B10 cells.Moreover,microorganisms can promote the expression of IL-10 of B10 cells,but do not change the total number of B10 cells.