發(fā)布時間：2018-07-02 15:08

  本文選題：原發(fā)性肝癌 + Vγ9Vδ2T細胞�。� 參考：《山西醫(yī)科大學(xué)》2016年碩士論文

【摘要】：目的:檢測HCC患者外周血的Vγ9Vδ2T細胞比例,分化成熟分型,免疫殺傷功能與健康人差別,體外擴增的效果及上述指標的變化。方法:采集HCC病人(n=25)和健康捐獻者(n=20)外周血共10 ml,采用不連續(xù)密度梯度離心法分離單核細胞層。1.應(yīng)用流式細胞術(shù)檢測Vγ9Vδ2T細胞占總T細胞比例,檢測分化成熟亞型,分泌IFN-γ和表達CD107a的細胞比例。2.分離的淋巴細胞加入GT-T551淋巴細胞培養(yǎng)基,唑來膦酸和IL-2加入培養(yǎng)基,37℃,5%CO2培養(yǎng)箱中培養(yǎng),2-3d定期加入IL-2,12-14d后收獲細胞。3.再次應(yīng)用流式細胞術(shù)檢測Vγ9Vδ2T細胞占總T細胞比例,分化成熟亞型,分泌IFN-γ和表達CD107a的細胞比例。結(jié)果:1.健康人組18例擴增成功,2例細胞擴增失敗�；颊呓M21例擴增成功,4例細胞擴增失敗。2.培養(yǎng)前HCC患者外周血中Vγ9Vδ2T細胞占總T細胞比例低于健康人,分化成熟比例在培養(yǎng)前與健康人相比無差異,分泌IFN-γ和表達CD107a比例與健康人相比無差異。3.體外擴增培養(yǎng)后HCC患者與健康人外周血中Vγ9Vδ2T細胞占總T細胞比例明顯升高。分化成熟亞型與擴增前對比發(fā)現(xiàn),培養(yǎng)后Tn、Tcm、Temra的比例較擴增前下降,患者和健康人Tem,Tem+Temra(總效應(yīng)型)明顯升高。擴增后HCC患者和健康人外周血中Vγ9Vδ2T細胞分泌IFN-γ和表達CD107a比例與擴增前無差異。4.擴增后HCC患者外周血Vγ9Vδ2T細胞比例,分化成熟亞型,分泌IFN-γ和表達CD107a比例與健康人對比無差異。結(jié)論:1.HCC患者外周血Vγ9Vδ2T細胞比例低于健康人,分化成熟亞型的分布與健康人相似,分泌IFN-γ和表達CD107a比例與健康人無差異。2.健康人和肝癌患者PBMCs經(jīng)過體外加入唑來膦酸+IL-2刺激,都可高度擴增Vγ9Vδ2T細胞。擴增后分化成熟亞型中總效應(yīng)細胞型明顯升高,分泌IFN-γ和表達CD107a比例無差異。3.這種方法可以為下一步過繼細胞免疫治療和實驗研究和臨床應(yīng)用提供合適的Vγ9Vδ2T細胞。
[Abstract]:Aim: to detect the percentage of V 緯 9V 未 2T cells in peripheral blood of patients with HCC, the differentiation and maturation type, the difference of immune killing function between HCC patients and healthy people, the effect of amplification in vitro and the changes of the above indexes. Methods: peripheral blood samples from patients with HCC (nong25) and healthy donors (nong20) were collected and the monocyte layer was isolated by discontinuous density gradient centrifugation. Flow cytometry was used to detect the percentage of V 緯 9V 未 2T cells to total T cells, to detect the mature subtypes of differentiation, to secrete IFN- 緯 and to express CD107a. The isolated lymphocytes were added to the GT-T551 lymphocyte culture medium. Zoledronic acid and IL-2 were added to the culture medium at 37 鈩，

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