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《臨床醫(yī)學(xué)論文-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.doc》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在工程資料-天天文庫(kù)。
1、臨床醫(yī)學(xué)論文■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(guò)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(yáng)性和雙陽(yáng)性分選法分別富集CD4+CD25+Trcg細(xì)胞和CD8+CTL；通過(guò)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(guò)TznsWcll分隔
2、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)體系屮加入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2.0X105/ml的CTL與l?0X105/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)體系屮以混合培養(yǎng)的方式加入質(zhì)量濃度為5.0X104/mlCD4+CD25+Treg細(xì)胞即可明顯抑制培養(yǎng)體系屮CTL對(duì)腫瘤細(xì)胞的
3、殺傷作汕如以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22.0X105/ml。結(jié)論CD4+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)制屮，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]ObjectiveToinvestigatetheeffectofe
4、nrichedCD4+CD25+TregulatorycellsoncytotoxicityofCTLagainstgastriccancercelllineMNK45cellsinvitroandexplorethepossibleunderlyingmechanism.MethodsAfterTcellsenrichment,MACSseparatingsystemwasusedtosortCD8+CTLandCD4+CD25+TregfromspleenTlymphocytes?TheoptimalconcentrationofCTLandt
5、hebestcultureconditionwereexploredbymeasuringthecytotoxicityofCTLagainstMNK45atdifferentdoses.ThenCD4+CD25+Tregwereaddedintotheculturesystemorinaseparatewc11intranswcl1system?TheinfluenceofTrcgoncytotoxicityofCTLwasinvestigatedbycountinginactiveMNK45cells?Mowovet,thecellularan
6、dimmunologicalmechanismwhichmightbeinvolvedwasanalyzed.ResultsTntheculturesystem,CTLshowedsignificanteffectagainstMNK45(P<0.01).When1.0ml(5.OX104/ml)wasaddedtothisreactionsystem,theamountofkiliedMNK45cellsdecreaseobviously(P<0.05)?AndwhenTregcelIswereculturedseperatlyintheTran
7、sWei1system,Tregcellsmustbeincreased2.0X105/mltoachievethesameinhibitoryeffect?ConclusionsCD4+CD25+TregulatorycellscouldexertinhibitoryeffectonCD8+CTLki1lingfunctioninvitro,thecelltocellcontactmechanismplaysimportantrolesinmediatingthiseffect?[Keywords]Gastrictumor;RegulatoryT
8、cells;CytotoxicTlymphocyte;MNK45止常機(jī)體的抗腫瘤機(jī)制主要包括體液免疫和細(xì)胞免疫兩方面，兩者
