膠質(zhì)瘤光動力疫苗的體外細胞毒性實驗研究

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1、·80·中國微侵襲神經(jīng)外科雜志2015年02月20日第20卷第2期ChinJMinimInvasiveNeurosurg,VOL.20,NO.2,February20,2015·實驗研究·膠質(zhì)瘤光動力疫苗的體外細胞毒性實驗研究袁開長，楊非，趙飛，林彬，李方成【摘要】目的制備膠質(zhì)瘤光動力疫苗并檢測體外抗腫瘤效應(yīng)。方法采用4種不同方法制備膠質(zhì)瘤疫苗。凍融疫苗：凍融法制備人膠質(zhì)瘤細胞U251抗原，呈遞給樹突狀細胞（DC）制備疫苗。熱休克疫苗：熱休克法處理人膠質(zhì)瘤細胞U251并提取抗原，呈遞給DC制備疫苗。光動力全細

2、胞抗原疫苗：光動力法提取人膠質(zhì)瘤細胞U251全細胞抗原并呈遞給DC制備疫苗。光動力洗脫抗原疫苗：光動力法處理人膠質(zhì)瘤細胞U251后，再用弱酸洗脫法提取活細胞表面抗原，呈遞給DC制備疫苗。將上述4種不同方法制備的疫苗在體外與自體淋巴細胞以及靶細胞U251反應(yīng)，然后用MTT法檢測靶細胞凋亡率及壞死率，比較4種疫苗的體外抗腫瘤細胞效應(yīng)。結(jié)果凍融疫苗、熱休克疫苗、光動力全細胞抗原疫苗和光動力洗脫抗原疫苗的靶細胞死亡率分別是：（10.02±0.32）%、（18.34±0.46）%、（35.56±1.75）%和（54.4

3、5±2.14）%，4組疫苗抗腫瘤細胞效應(yīng)是光動力洗脫抗原疫苗＞光動力全細胞抗原疫苗＞熱休克疫苗＞凍融疫苗。結(jié)論膠質(zhì)瘤光動力洗脫抗原疫苗是一種具有良好體外抗腫瘤效應(yīng)的新型疫苗?！娟P(guān)鍵詞】神經(jīng)膠質(zhì)瘤；化學療法；疫苗；光動力；細胞毒性中圖分類號：R730.264文獻標志碼：Adoi：10.11850/j.issn.1009-122X.2015.02.012Invitrocytotoxicityofgliomaphotodynamicvaccine:anexperimentalstudyYuanKaizhang1,Y

4、angFei1,ZhaoFei1,LinBin1,LiFangcheng21.DepartmentofNeurosurgery,theThirdAffiliatedHospitalofAnhuiMedicalUniversity,HefeiBinhuHospital,Hefei,Anhui230601,China;2.DepartmentofNeurosurgery,theSecondAffiliatedHospitalofSunYat-SenUniversity,Guangzhou,Guangdong510

5、120,ChinaAbstract:ObjectiveTopreparegliomaphotodynamicvaccinesanddetecttheiranti-tumoreffectsinvitro.MethodsFourkindsofgliomaspecifictumorvaccineswerepreparedbydifferentmethods.Freeze-thawcellvaccine(No.1):whole-cellantigensofhumangliomacellslineU251wereext

6、ractedusingfreeze-thawmethodtoproducedendriticcell(DC)vaccine.Heatshocktumorvaccine(No.2):antigensofhumangliomacelllineU251wereextractedafterheatshockprocessing,whichwerepresentedtoDCforvaccines.Photodynamicwhole-cellantigentumorvaccine(No.3):humangliomacel

7、llineU251cellsasantigenwerepresentedtoDCtoproducevaccinesbyphotodynamicextractionmethod.Photodynamicelutionantigentumorvaccine(No.4):U251cellswerecultivatedwithphotosensitizersand,aftertreatmentbyphotodynamicmethod,elutedbyweakacidsolutiontoextractantigensf

8、orpreparingDCvaccine.Tocomparetheeffectsofanti-cancerinvitro,the4vaccinesweremixedandreactedrespectivelywithhostlymphocytesandU251cells,andtheapoptosisandnecrosisratesoftheU251cellsweredetectedbyMTTass