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《體外富集treg細(xì)胞對(duì)ctl殺傷mnk45胃癌細(xì)胞活性的影響及機(jī)制研究》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫。
1、體外富集Treg細(xì)胞對(duì)CTL殺傷MNK45胃癌細(xì)胞活性的影響及機(jī)制研究作者：曾冬竹鄭峻松王自強(qiáng)吳淼余佩武【摘要】目的體外分析CD4+CD25+Treg(regulatoryTcells)細(xì)胞對(duì)CD8+CTL(cytotoxicTlymphocytes)細(xì)胞殺傷MNK45胃癌細(xì)胞株活性的影響及機(jī)制。方法通過T細(xì)胞純化柱分選小鼠脾細(xì)胞懸液T淋巴細(xì)胞，以MACS單陽性和雙陽性分選法分別富集CD4+CD25+Treg細(xì)胞和CD8+CTL；通過不同濃度CD8+CTL與MNK45胃癌細(xì)胞混合培養(yǎng)，篩選有效殺瘤體外培養(yǎng)體系。通過TransWell分
2、隔培養(yǎng)和混合培養(yǎng)的方式向CTL與MNK45胃癌細(xì)胞株組成的體外抗腫瘤培養(yǎng)體系中加入CD4+CD25+Treg細(xì)胞，以瑞氏染色顯微計(jì)數(shù)的方法觀察Treg細(xì)胞對(duì)CTL殺傷MNK45胃癌細(xì)胞活性的影響，結(jié)合ELISA法檢測(cè)各體系穿孔素濃度，分析影響活性的細(xì)胞及免疫學(xué)機(jī)制。結(jié)果質(zhì)量濃度為2.0×105/ml的CTL與1.0×105/mlMNK45胃癌細(xì)胞混合培養(yǎng)，CTL能對(duì)MNK45胃癌細(xì)胞形成明顯的殺傷作用(P<0.01)；在此培養(yǎng)體系中以混合培養(yǎng)的方式加入質(zhì)量濃度為5.0×104/mlCD4+CD25+Treg細(xì)胞即可明顯抑制培養(yǎng)體
3、系中CTL對(duì)腫瘤細(xì)胞的殺傷作用；如以TransWell分隔培養(yǎng)的方式將CD4+CD25+Treg細(xì)胞與抗腫瘤培養(yǎng)體系分隔培養(yǎng)，達(dá)到同樣抑制效能(實(shí)驗(yàn)組死亡腫瘤細(xì)胞百分率與對(duì)照組比較，P<0.05)的CD4+CD25+Treg細(xì)胞濃度須≥2.0×105/ml。結(jié)論13CD4+CD25+Treg細(xì)胞能夠?qū)TL殺傷MNK45胃癌細(xì)胞株的活性形成明顯抑制，在Treg細(xì)胞與CTL作用的相應(yīng)細(xì)胞和免疫學(xué)機(jī)制中，細(xì)胞接觸機(jī)制占主導(dǎo)地位?！娟P(guān)鍵詞】胃腫瘤調(diào)節(jié)性T細(xì)胞細(xì)胞毒性T淋巴細(xì)胞MNK45胃癌細(xì)胞株【Abstract】ObjectiveT
4、oinvestigatetheeffectofenrichedCD4+CD25+TregulatorycellsoncytotoxicityofCTLagainstgastriccancercelllineMNK45cellsinvitroandexplorethepossibleunderlyingmechanism.MethodsAfterTcellsenrichment,MACSseparatingsystemwasusedtosortCD8+CTLandCD4+CD25+TregfromspleenTlymphocytes.T
5、heoptimalconcentrationofCTLandthebestcultureconditionwereexploredbymeasuringthecytotoxicityofCTLagainstMNK45atdifferentdoses.ThenCD4+CD25+Tregwereaddedintotheculturesystemorinaseparatewellintranswellsystem.TheinfluenceofTregoncytotoxicityofCTLwasinvestigatedbycountingin
6、activeMNK45cells.Moreover,thecellularandimmunologicalmechanismwhichmightbeinvolvedwasanalyzed.ResultsIntheculturesystem,CTLshowedsignificanteffectagainstMNK45(P<0.01).When1.0ml(5.0×104/ml)wasaddedtothis13reactionsystem,theamountofkilledMNK45cellsdecreaseobviously(P&l
7、t;0.05).AndwhenTregcellswereculturedseperatlyintheTransWellsystem,Tregcellsmustbeincreased2.0×105/mltoachievethesameinhibitoryeffect.ConclusionsCD4+CD25+TregulatorycellscouldexertinhibitoryeffectonCD8+CTLkillingfunctioninvitro,thecelltocellcontactmechanismplaysimporta
8、ntrolesinmediatingthiseffect.【Keywords】Gastrictumor;RegulatoryTcells;CytotoxicTlymphocyte;MNK45正常機(jī)體的抗腫瘤機(jī)制主要包括體
