發(fā)布時(shí)間：2018-04-21 14:09

  本文選題：樹突狀細(xì)胞 + 體外誘導(dǎo)��； 參考：《細(xì)胞與分子免疫學(xué)雜志》2015年12期

【摘要】：目的探討并比較不同細(xì)胞因子組合、誘導(dǎo)時(shí)間對(duì)樹突狀細(xì)胞(DC)體外誘導(dǎo)、成熟,分泌細(xì)胞因子水平和刺激淋巴細(xì)胞增殖能力的影響,為建立DC體外高效、規(guī)�；嘤w系提供依據(jù)。方法分離健康人外周血單個(gè)核細(xì)胞(PBMC),貼壁法獲得DC前體細(xì)胞,用白細(xì)胞介素4(IL-4)和粒細(xì)胞巨噬細(xì)胞集落刺激因子(GM-CSF)誘導(dǎo)2 d,將其分為6組。A組加入腫瘤壞死因子α(TNF-α)、IL-1α和前列腺素E2(PGE2);B組加入α干擾素(IFN-α);C組加入脂多糖(LPS);3組細(xì)胞繼續(xù)培育2 d。D~F組細(xì)胞先進(jìn)行1/3體積換液,24 h后,分別再加入上述A~C組的成熟因子,再繼續(xù)誘導(dǎo)2 d。分別收獲各組上清和細(xì)胞,進(jìn)行細(xì)胞計(jì)數(shù),采用流式細(xì)胞術(shù)進(jìn)行表型分析;采用ELISA檢測(cè)各組DC分泌IL-12p70的水平、采用細(xì)胞增殖檢測(cè)試劑盒刺激淋巴細(xì)胞增殖的能力。結(jié)果 6組體外誘導(dǎo)體系均能培育出形態(tài)學(xué)類似DC的細(xì)胞,DC數(shù)量與純度(均達(dá)到96%以上),各組間無顯著性差異。A組與D組比較:CD40、CD80、CD83、CD86共刺激分子表達(dá)水平相近,但D組DC分泌IL-12p70水平高于A組;B組與E組比較:E組DC的CD86表達(dá)水平顯著高于B組,CD80表達(dá)量略高于B組,2組DC分泌IL-12p70的水平相近;C組與F組比較:CD40、CD80、CD83、CD86共刺激分子表達(dá)水平和分泌IL-12p70的水平均相近。6組DC刺激淋巴細(xì)胞增殖水平均相近。三種不同成熟因子(組合)比較:B組DC共刺激分子CD80水平明顯低于C組和A組,且IL-12p70分泌水平最低,而C組和A組共刺激分子表達(dá)水平與IL-12p70分泌水平相近。結(jié)論 TNF-α、IL-1α和PGE-2三種細(xì)胞因子聯(lián)合以及LPS可有效地誘導(dǎo)DC表達(dá)高水平免疫共刺激分子、分泌高水平IL-12p70。
[Abstract]:Objective to explore and compare the effects of different cytokine combinations, induction time on the induction, maturation, secretion of cytokine levels and stimulation of lymphocyte proliferation in vitro of dendritic cells (DC), in order to provide a basis for the establishment of an efficient and large-scale culture system for DC in vitro, and to separate the peripheral blood mononuclear cells (PBMC) of healthy people before DC. Somatic cells were induced by interleukin 4 (IL-4) and granulocyte macrophage colony stimulating factor (GM-CSF) to induce 2 D, which were divided into 6 groups of.A groups with tumor necrosis factor alpha (TNF- alpha), IL-1 alpha and prostaglandin E2 (PGE2), B group adding interferon alpha (IFN- a), C group adding lipoprotein (LPS), and the 3 groups continued to cultivate 2 groups of cells to replace the volume of liquid first, After 24 h, the mature factors of the A~C group were added, and then 2 d. were further induced to harvest the supernatant and cells, the cell count, the flow cytometry was used for phenotypic analysis, the level of DC secretion of IL-12p70 was detected by ELISA, and the ability to stimulate the lymphocyte proliferation by the cell proliferation detection kit was used. The results of the 6 groups were induced in vitro. The number and purity of DC and purity (all up to 96%) could be produced in all conductors, and there was no significant difference between groups.A and D group: CD40, CD80, CD83, CD86 co stimulator level was similar, but D group DC secreted IL-12p70 level higher than that of A group. The level of DC secreted IL-12p70 in 2 groups was similar, and in group C and F group, CD40, CD80, CD83, CD86 co stimulatory molecules expression level and IL-12p70 level were similar to those of.6 group. The comparison of three different mature factors (combination) was significantly lower than that of the group and group. The secretory level of 12p70 is the lowest, while the expression level of CO stimulatory molecules in group C and A is similar to that of IL-12p70 secreting. Conclusion TNF- alpha, IL-1 alpha and PGE-2 three cytokine combinations and LPS can effectively induce high level immune costimulatory molecules in DC expression, and secrete high level IL-12p70..

【作者單位】： 南京市胸科醫(yī)院胸外科;南京市免疫細(xì)胞生物工程技術(shù)研究中心;南京市胸科醫(yī)院呼吸內(nèi)科;
【分類號(hào)】：R329.2

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