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

再胺球蛋白血症是一种导致缺乏特定免疫球蛋白的免疫缺陷疾病。在目前的研究中，我们检测了aly/aly小鼠的T细胞功能作为无球蛋白血症的模型，小鼠显示出严重的联合免疫缺陷，包括缺乏IgA和IgG的合成。aly/aly小鼠也缺少Peyer's补丁。组织学检查显示胸腺髓质和皮质边缘不清。与常规小鼠相比，脾脏中卵泡数量减少。从脾脏中分离淋巴细胞，用流式细胞仪测定T/B细胞比。Aly/ Aly小鼠T/B细胞比例正常，但总恢复细胞数明显低于常规小鼠。对aly/aly小鼠进行体内和体外抗原刺激均未发现抗体应答。RT-PCR检测T细胞的细胞因子谱。我们发现在aly/aly小鼠中IL-2 mRNA的合成水平很低，而其他细胞因子的mRNA合成水平通过RT-PCR不断检测。IL-2受体在T细胞中的表达也处于低水平。通过抗cd3处理的细胞增殖试验、ELISA和FACS分析证实了这种il - 2缺陷。CD3和PMA刺激T细胞后，集落形成的大小和数量减少。体外抗原刺激实验中加入il - 2后，IgM抗体反应恢复。混合淋巴细胞培养实验显示低水平的B细胞反应。T细胞克隆的建立没有成功。提示aly/aly小鼠无球蛋白血症可能是由T细胞中IL2合成缺陷引起的。然而，B细胞功能也参与了这种免疫缺陷。需要进一步的分子碱基分析来解决双球蛋白血症中的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.