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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。
Objective To analyze the effects of age, gender and environmental factors on B10 cells in spleen of Balb / c mice. Methods The mice were divided into groups by age, gender and environment. The proportion of CD5 + CD1dhiB cells in spleens of mice was analyzed by flow cytometry. The mouse splenocytes were cultured in vitro, and the proportion of CD19 + IL-10 + B cells was analyzed by flow cytometry without any stimulus or stimulation with LPS + PIM for 5 h. Results The average percentage of CD5 + CD1dhiB cells in spleens of 6-month-old mice was 2.9%, significantly higher than that of 2-month-old mice (2.28%, P <0.05). Without any stimulation, The percentage of IL-10 + B cells in spleen of 6-month-old mice averaged 1.93% after adding LPS + PIM for 5 h , Higher than 1.25% of 2-month-old mice (P <0.05). Cell phenotype and intracellular IL-10 staining analysis showed no significant difference in age-matched female and male spleen B10 cells (P> 0.05). There was no significant difference in the proportion of IL-10 + B cells induced by LPS + PIM between normal and SPF mice CD5 + CD1dhiB cells (P> 0.05), while no stimulation of normal mice spleen IL- The average percentage of 10 + B cells was 0.87%, which was significantly higher than that of SPF mice (0.38%, P <0.01). Conclusion The splenic B10 cells can be expanded with the age of Balb / c mice. The expression of B10 cells has no significant relationship with sex differences. The microbial environment can promote the activation of B10 cells but has no obvious effect on the total number of B10 cells.