Analysis of T cell function in Agammaglobulinemia-aly/aly mice-

无氨基球蛋白血症是一种严重的免疫缺陷疾病，导致缺乏特定的免疫球蛋白。在目前的研究中，我们已经检查了作为无丙种球蛋白血症模型的aly/aly小鼠的T细胞功能，该小鼠显示出严重的联合免疫缺陷，包括缺乏伊加和IgG合成。aly/aly小鼠也缺乏派伊尔集合淋巴结。组织学检查胸腺髓质、皮质边界不清。与常规小鼠相比，脾脏中的卵泡数量减少。从脾中分离淋巴细胞并通过FACS测量T/B细胞比率。Aly/aly小鼠显示正常的T/B细胞比率，但是总回收的细胞数目明显低于常规小鼠。aly/aly小鼠的体内和体外抗原刺激未显示抗体应答。用RT-PCR检测T细胞的细胞因子谱，发现aly/aly小鼠IL-2 mRNA的合成量较低，而其它细胞因子mRNA的合成量持续增加，T细胞IL-2受体的表达量也较低。通过抗-CD 3处理的细胞增殖测定、ELISA和FACS分析证实了这种IL 2缺陷。CD 3和PMA刺激T细胞后，集落形成的大小和数量减少。在体外抗原刺激试验中加入IL 2后，IgM抗体应答恢复。混合淋巴细胞培养实验显示低水平的B细胞反应。T细胞克隆建立失败。结果提示，aly/aly小鼠无丙种球蛋白血症可能是由于T细胞合成IL 2不足所致。然而，B细胞功能也参与了这种免疫缺陷。需要进一步的分子基础分析来解决无丙种球蛋白血症中的T细胞功能障碍。

Agaminaglobulinemia is serve immunodeficient disease that resulted in a lack of particular immunoglobulins. In current study, we have examined T cell function of aly/aly mice as a model of agammaglobulinemia that the mice revealed a severe combined immunodificiency including a lack of IgA and IgG synthesis. The aly/aly mice are also a lack of Peyer's patches. On the histological examination, the mice revealed unclear margin of medulla and cortex in thymus. The number of follicles in the spleen was reduced in compared with conventional mice. The lymphocytes were isolated from spleen and measured T/B cell ratio by FACS.Aly/aly mice showed normal T/B cell ratios however the total recovered cell numbers were clearly low than conventional mice. In vivo and In vitro antigen stimulation of the aly/aly mice revealed no antibody response. The cytokine profiles of T cells were examined by RT-PCR.We found that IL-2 mRNA synthesis was quite low revels in the aly/aly mice, while other cytokines mRNA syntheses were constantly detected by RT-PCR.The IL-2 receptor expression in T cell was also in low levels. This IL2 deficiency was proved by an anti-CD3 treated cell proliferation assay, ELISA and FACS analysis. The size and number of colony formation after CD3 and PMA stimulation of T cells were decreased. IgM antibody response was recovered after IL2 adding to in vitro antigen stimulation assay. An experiment of mix lymphocyte culture revealed low levels of B cell response. The establishment of T cell clone was not succeeded. The result indicated that agammaglobulinemia of the aly/aly mice may be caused by the deficient of IL2 synthesis in T cells. However, B cell function was also involved in this immunodeficiency. Farther molecular base analysis was required for resolve T cell dysfunction in an agammaglobulinemia.