安徽医科大学学报
安徽醫科大學學報
안휘의과대학학보
ACTA UNIVERSITY MEDICINALIS ANHUI
2015年
1期
4-8
,共5页
赵皓%余晾%张敬星%陈月娟%吕合作%王保龙
趙皓%餘晾%張敬星%陳月娟%呂閤作%王保龍
조호%여량%장경성%진월연%려합작%왕보룡
髓鞘碱性蛋白%淋巴细胞%免疫%流式细胞术
髓鞘堿性蛋白%淋巴細胞%免疫%流式細胞術
수초감성단백%림파세포%면역%류식세포술
myelin basic protein%lymphocytes%immunity%flow cytometry
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。
目的:培養針對髓鞘堿性蛋白( MBP)特異的SD大鼠T淋巴細胞,對其錶型和細胞因子分泌特徵進行鑒定。方法將MBP免疫SD大鼠,收集迴流淋巴結,製備成單細胞懸液,在RMPI 1640培養液中用MBP反複刺激穫得MBP特異性T細胞(MBP-T)。然後用3H-胸腺嘧啶覈苷(3H-TdR)摻入法測定其對MBP刺激的反應性,用流式細胞術鑒定其錶型和細胞因子的產生。結果噹 MBP抗原存在的情況下,MBP-T的3 H-TdR摻入量明顯增加。流式細胞術檢測錶明MBP-T主要為CD3+ CD4+的T淋巴細胞(98%),併可以產生榦擾素γ(IFN-γ)和白細胞介素10(IL-10)等細胞因子。結論 MBP免疫SD大鼠的迴流淋巴結細胞,經過體外擴增可以穫得 MBP 特異性的 T 細胞,其錶型主要為 CD3+CD4+,併可以產生Th1和Tr1型細胞因子。
목적:배양침대수초감성단백( MBP)특이적SD대서T림파세포,대기표형화세포인자분비특정진행감정。방법장MBP면역SD대서,수집회류림파결,제비성단세포현액,재RMPI 1640배양액중용MBP반복자격획득MBP특이성T세포(MBP-T)。연후용3H-흉선밀정핵감(3H-TdR)참입법측정기대MBP자격적반응성,용류식세포술감정기표형화세포인자적산생。결과당 MBP항원존재적정황하,MBP-T적3 H-TdR참입량명현증가。류식세포술검측표명MBP-T주요위CD3+ CD4+적T림파세포(98%),병가이산생간우소γ(IFN-γ)화백세포개소10(IL-10)등세포인자。결론 MBP면역SD대서적회류림파결세포,경과체외확증가이획득 MBP 특이성적 T 세포,기표형주요위 CD3+CD4+,병가이산생Th1화Tr1형세포인자。
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.