鞘碱性蛋白活化的SD大鼠T细胞培养与鉴定

目的：培养针对髓鞘碱性蛋白( MBP)特异的SD大鼠T淋巴细胞,对其表型和细胞因子分泌特征进行鉴定。方法将MBP免疫SD大鼠,收集回流淋巴结,制备成单细胞悬液,在RMPI 1640培养液中用MBP反复刺激获得MBP特异性T细胞(MBP-T)。然后用3H-胸腺嘧啶核苷(3H-TdR)掺入法测定其对MBP刺激的反应性,用流式细胞术鉴定其表型和细胞因子的产生。结果当 MBP抗原存在的情况下,MBP-T的3 H-TdR掺入量明显增加。流式细胞术检测表明MBP-T主要为CD3+ CD4+的T淋巴细胞(98%),并可以产生干扰素γ(IFN-γ)和白细胞介素10(IL-10)等细胞因子。结论 MBP免疫SD大鼠的回流淋巴结细胞,经过体外扩增可以获得 MBP 特异性的 T 细胞,其表型主要为 CD3+CD4+,并可以产生Th1和Tr1型细胞因子。

Culture and identification of myelin basic protein activated T cells in SD rats

Objective To culture the myelin basic protein ( MBP) specific T lymphocytes in SD rats, and identify their characterization of phenotype and cytokine profiles. Methods The SD rats were immunized with MBP; the draining lymph nodes were collected and made into single cell suspension. The cells were cultured in RMPI 1640 medium and stimulated repeatedly with MBP to produce MBP specific T ( MBP-T) cells. Then, their reactivity to MBP stimulation was measured using 3 H-thymidine ( 3 H-TdR) incorporation, the phenotype and cytokine profiles were determined using flow cytometry. Results When the MBP antigen exists, 3 H-TdR incorporation of MBP-T cells increased obviously. Flow cytometry showed that MBP-T cells were mainly CD3 + CD4 + T lymphocytes (98%), and could produce interferon-γ(IFN-γ) and interleukin-10(IL-10). Conclusion The cells collected from the draining lymph nodes of MBP immunized SD rats could be amplified into MBP-T cells in vitro. The pheno-types of these cells are mainly CD3 + CD4 +, and show the Th1 and Tr1 cytokine profiles.