摘要
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3H-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。
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 ^3H-thymidine （ ^3H-TdR） incorporation, the phenotype and cytokine profiles were determined using flow cytometry. Results When the MBP antigen exists, ^3H-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.
出处
《安徽医科大学学报》
CAS
北大核心
2015年第1期4-8,共5页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:81071268、81271363)
