Article(id=1222469707249213691, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1222469705873481976, articleNumber=null, orderNo=null, doi=10.16438/j.0513-4870.2019-0670, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1566403200000, receivedDateStr=2019-08-22, revisedDate=1567699200000, revisedDateStr=2019-09-06, acceptedDate=null, acceptedDateStr=null, onlineDate=1769389090596, onlineDateStr=2026-01-26, pubDate=1570809600000, pubDateStr=2019-10-12, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1769389090596, onlineIssueDateStr=2026-01-26, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1769389090596, creator=13701087609, updateTime=1769389090596, updator=13701087609, issue=Issue{id=1222469705873481976, tenantId=1146029695717560320, journalId=1189982191388893191, year='2019', volume='54', issue='10', pageStart='1711', pageEnd='1880', issueExtLink='null', onlineDate='null', pubDate='1570809600000', pubDateStr='2019-10-12', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1769389090269, creator='13701087609', updateTime=1769389551199, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1222471639254683958, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1222469705873481976, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1222471639254683959, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1222469705873481976, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=1863, endPage=1867, ext={EN=ArticleExt(id=1222469707685421313, articleId=1222469707249213691, tenantId=1146029695717560320, journalId=1189982191388893191, language=EN, title=The effects of different stimulators on the activation of human lymphocytes, columnId=1190335348761793317, journalTitle=Acta Pharmaceutica Sinica, columnName=Original Articles, runingTitle=null, highlight=null, articleAbstract=

The research aims to study the effects of different stimulants on the activation of human lymphocytes. Human peripheral blood mononuclear cells were prepared by density centrifugation. The blood's sample was provided by National Institutes for Food and Drug Control and approved by its Ethics Committee. The expressions of CD69 in CD3+CD4+ and CD3+CD8+ human T cells were detected by flow cytometry after administrated with CD3/CD28 antibody, phytohaemagglutinin (PHA), Staphylococcus auresus enterooxin B (SEB), interleukin (IL27) and PMA plus ionomycin for 24 h. The proliferation of lymphocyte was detected by CellTiter-Glo kit. The secreted IFNγ in supernatant of medium was examined by ELISA kit. The proliferation of lymphocytes had no change after exposed of CD3/CD28 antibody, SEB, IL27 and PMA plus ionomycin for 24 h. However, the CD69 expressions in CD3+CD4+ and CD3+CD8+ T cells and IFNγ productions were significantly increased by CD3/CD28 antibody, SEB, IL27 and PMA plus ionomycin at 24 h, indicating that CD3+CD4+ and CD3+CD8+ T cells were activated under above-mentioned stimulated condition. CD3/CD28 antibody, SEB, IL27 and PMA plus ionomycin were valid stimulants for T cell activation.

, authors=null, authorsList=Zheng YAN, Ni-na XUE, Ming JI, Fang-fang LAI, Xiao-guang CHEN, authorCompany=null, correspAuthors=Xiao-guang CHEN, authorNote=null, correspAuthorsNote=null, copyrightStatement=Copyright ©2019 Acta Pharmaceutica Sinica. All rights reserved., copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1222469708620751140, articleId=1222469707249213691, tenantId=1146029695717560320, journalId=1189982191388893191, language=CN, title=不同刺激剂对人淋巴细胞活化的影响, columnId=1190335348896011050, journalTitle=药学学报, columnName=研究论文, runingTitle=null, highlight=null, articleAbstract=

本文研究不同刺激剂在体外对人淋巴细胞活化的影响。采用梯度离心法分离获得人外周血单个核细胞(peripheral blood mononuclear cells,PBMC),血液样本由中国食品药品检定研究院提供,并通过伦理委员会批准。用CD3/CD28抗体(anti-CD3/CD28 antibody)、植物血凝素(phytohaemagglutinin,PHA)、金黄色葡萄球菌B型肠毒素(Staphylococcus auresus enterooxin B,SEB)、白介素27(interleukin 27,IL27)、佛波酯(phorbol myristrate acetate,PMA)+离子霉素(ionomycin)在体外分别刺激24 h,观察CD3+CD4+和CD3+CD8+ T细胞亚群CD69表达。同时采用CellTiter-Glo发光法和ELISA法检测淋巴细胞增殖和IFNγ的分泌。CD3/CD28抗体、PMA+ionomycin、SEB和IL27作用PBMC 24 h不影响淋巴细胞增殖,但可以很好地活化CD3+CD4+和CD3+CD8+ T细胞,表现为CD69的平均荧光强度显著右移,并促进淋巴细胞IFNγ的分泌。CD3/CD28抗体、PMA+ionomycin、SEB和IL27是T细胞活化的有效刺激剂。

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*陈晓光, Tel:86-10-63165207, E-mail:
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tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, language=CN, orderNo=4, keyword=淋巴细胞活化)], refs=[Reference(id=1222469711934251412, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, doi=10.1038/s41577-018-0044-0, pmid=null, pmcid=null, year=2018, volume=18, issue=null, pageStart=635, pageEnd=647, url=null, language=null, rfNumber=[1], rfOrder=0, authorNames=Borst J, Ahrends T, Bąbała N, journalName=Nat Rev Immunol, refType=null, unstructuredReference= Borst J , Ahrends T , Bąbała N et al . CD4+ T cell help in cancer immunology and immunotherapy[J]. Nat Rev Immunol, 2018, 18: 635-647., articleTitle=CD4+ T cell help in cancer immunology and immunotherapy, refAbstract=null), Reference(id=1222469711997165975, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, doi=null, pmid=null, pmcid=null, year=2014, volume=5, issue=null, pageStart=12472, pageEnd=12508, url=https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4350348/, language=null, rfNumber=[2], rfOrder=1, authorNames=Galluzzi L, Vacchelli E, Bravo-San Pedro JM, journalName=Oncotarget, refType=null, unstructuredReference= Galluzzi L , Vacchelli E , Bravo-San Pedro JM et al . Classification of current anticancer immunotherapies[J]. Oncotarget, 2014, 5: 12472-12508., articleTitle=Classification of current anticancer immunotherapies, refAbstract=null), Reference(id=1222469712076857753, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, doi=10.1016/j.coi.2017.07.004, pmid=null, pmcid=null, year=2017, volume=47, issue=null, pageStart=85, pageEnd=92, url=null, language=null, rfNumber=[3], rfOrder=2, authorNames=Melssen M, Slingluff CL Jr, journalName=Curr Opin Immunol, refType=null, unstructuredReference= Melssen M , Slingluff CL Jr . Vaccines targeting helper T cells for cancer immunotherapy[J]. Curr Opin Immunol, 2017, 47: 85-92., articleTitle=Vaccines targeting helper T cells for cancer immunotherapy, refAbstract=null), Reference(id=1222469712164938140, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, doi=10.1038/nri1413, pmid=null, pmcid=null, year=2004, volume=4, issue=null, pageStart=595, pageEnd=602, url=null, language=null, rfNumber=[4], rfOrder=3, authorNames=Bevan MJ, journalName=Nat Rev Immunol, refType=null, unstructuredReference= Bevan MJ . Helping the CD8+ T-cell response[J]. Nat Rev Immunol, 2004, 4: 595-602., articleTitle=Helping the CD8+ T-cell response, refAbstract=null), Reference(id=1222469712261407135, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, doi=10.1016/S0145-2126(00)00003-5, pmid=null, pmcid=null, year=2000, volume=24, issue=null, pageStart=445, pageEnd=452, url=null, language=null, rfNumber=[5], rfOrder=4, authorNames=Mollgard L, Tidefelt U, Sundman-Engberg B, journalName=Leuk Res, refType=null, unstructuredReference= Mollgard L , Tidefelt U , Sundman-Engberg B et al . In vitro chemosensitivity testing in acute non lymphocytic leukemia using the bioluminescence ATP assay[J]. Leuk Res, 2000, 24: 445-452., articleTitle=In vitro chemosensitivity testing in acute non lymphocytic leukemia using the bioluminescence ATP assay, refAbstract=null), Reference(id=1222469712366264738, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, doi=10.1016/0022-1759(84)90440-X, pmid=null, pmcid=null, year=1984, volume=72, issue=null, pageStart=127, pageEnd=132, url=null, language=null, rfNumber=[6], rfOrder=5, authorNames=Ishizaka A, Tono-oka T, Matsumoto S, journalName=J Immunol Methods, refType=null, unstructuredReference= Ishizaka A , Tono-oka T , Matsumoto S . Evaluation of the proliferative response of lymphocytes by measurement of intracellular ATP[J]. J Immunol Methods, 1984, 72: 127-132., articleTitle=Evaluation of the proliferative response of lymphocytes by measurement of intracellular ATP, refAbstract=null), Reference(id=1222469712462733734, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, doi=null, pmid=null, pmcid=null, year=2017, volume=33, issue=null, pageStart=129, pageEnd=134, url=http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=hljsxyxb201702001, language=null, rfNumber=[7], rfOrder=6, authorNames=Xue NN, Dong K, Lai FF, journalName=J Harbin Univ Commerce (Nat Sci Ed) (哈尔滨商业大学学报(自然科学版)), refType=null, unstructuredReference= Xue NN , Dong K , Lai FF et al . CFSE-labeled proliferative assays for assessment of T cell function induced by different stimulants[J]. J Harbin Univ Commerce (Nat Sci Ed) (哈尔滨商业大学学报(自然科学版)), 2017, 33: 129-134., articleTitle=CFSE-labeled proliferative assays for assessment of T cell function induced by different stimulants, refAbstract=null), Reference(id=1222469712563397034, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, doi=10.1016/S0531-5565(98)00058-8, pmid=null, pmcid=null, year=1999, volume=34, issue=null, pageStart=79, pageEnd=93, url=null, language=null, rfNumber=[8], rfOrder=7, authorNames=Rea IM, McNerlan SE, Alexander HD, journalName=Exp Gerontol, refType=null, unstructuredReference= Rea IM , McNerlan SE , Alexander HD . CD69, CD25, and HLA-DR activation antigen expression on CD3+ lymphocytes and relationship to serum TNF-α, IFN-γ, and sIL-2R levels in aging[J]. Exp Gerontol, 1999, 34: 79-93., articleTitle=CD69, CD25, and HLA-DR activation antigen expression on CD3+ lymphocytes and relationship to serum TNF-α, IFN-γ, and sIL-2R levels in aging, refAbstract=null), Reference(id=1222469712647283116, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, doi=10.1016/0167-5699(96)10057-C, pmid=null, pmcid=null, year=1996, volume=17, issue=null, pageStart=481, pageEnd=486, url=null, language=null, rfNumber=[9], rfOrder=8, authorNames=Theze J, Alzari PM, Bertoglio J, journalName=Immunol Today, refType=null, unstructuredReference= Theze J , Alzari PM , Bertoglio J . Interleukin 2 and its receptors:recent advances and new immunological functions[J]. Immunol Today, 1996, 17: 481-486., articleTitle=Interleukin 2 and its receptors:recent advances and new immunological functions, refAbstract=null), Reference(id=1222469712768917934, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, doi=10.1002/stem.5530120502, pmid=null, pmcid=null, year=1994, volume=12, issue=null, pageStart=456, pageEnd=465, url=null, language=null, rfNumber=[10], rfOrder=9, authorNames=Ziegler SF, Ramsdell F, Alderson MR, journalName=Stem Cells, refType=null, unstructuredReference= Ziegler SF , Ramsdell F , Alderson MR . The activation antigen CD69[J]. Stem Cells, 1994, 12: 456-465., articleTitle=The activation antigen CD69, refAbstract=null), Reference(id=1222469712844415410, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, doi=10.1016/S0022-1759(03)00010-3, pmid=null, pmcid=null, year=2003, volume=275, issue=null, pageStart=251, pageEnd=255, url=null, language=null, rfNumber=[11], rfOrder=10, authorNames=Trickett A, Kwan YL, journalName=J Immunol Methods, refType=null, unstructuredReference= Trickett A , Kwan YL . T cell stimulation and expansion using anti-CD3/CD28 beads[J]. J Immunol Methods, 2003, 275: 251-255., articleTitle=T cell stimulation and expansion using anti-CD3/CD28 beads, refAbstract=null), Reference(id=1222469712911524276, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, doi=10.1186/s13287-016-0348-6, pmid=null, pmcid=null, year=2016, volume=7, issue=null, pageStart=90, pageEnd=null, url=null, language=null, rfNumber=[12], rfOrder=11, authorNames=Seo HH, Lee CY, Lee J, journalName=Stem Cell Res Ther, refType=null, unstructuredReference= Seo HH , Lee CY , Lee J et al . The role of nuclear factor of activated T cells during phorbol myristate acetate-induced cardiac differentiation of mesenchymal stem cells[J]. Stem Cell Res Ther, 2016, 7: 90., articleTitle=The role of nuclear factor of activated T cells during phorbol myristate acetate-induced cardiac differentiation of mesenchymal stem cells, refAbstract=null), Reference(id=1222469713007993273, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, doi=10.1080/10715762.2016.1227074, pmid=null, pmcid=null, year=2016, volume=50, issue=null, pageStart=1214, pageEnd=1225, url=null, language=null, rfNumber=[13], rfOrder=12, authorNames=Nakamura S, Nakanishi A, Takazawa M, journalName=Free Radic Res, refType=null, unstructuredReference= Nakamura S , Nakanishi A , Takazawa M et al . Ionomycin-induced calcium influx induces neurite degeneration in mouse neuroblastoma cells:analysis of a time-lapse live cell imaging system[J]. Free Radic Res, 2016, 50: 1214-1225., articleTitle=Ionomycin-induced calcium influx induces neurite degeneration in mouse neuroblastoma cells:analysis of a time-lapse live cell imaging system, refAbstract=null), Reference(id=1222469713158988221, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, doi=10.3390/toxins7051821, pmid=null, pmcid=null, year=2015, volume=7, issue=null, pageStart=1821, pageEnd=1836, url=null, language=null, rfNumber=[14], rfOrder=13, authorNames=Xu SX, Kasper KJ, Zeppa JJ, journalName=Toxins (Basel), refType=null, unstructuredReference= Xu SX , Kasper KJ , Zeppa JJ et al . Superantigens modulate bacterial density during Staphylococcus aureus nasal colonization[J]. Toxins (Basel), 2015, 7: 1821-1836., articleTitle=Superantigens modulate bacterial density during Staphylococcus aureus nasal colonization, refAbstract=null), Reference(id=1222469713238679999, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, doi=null, pmid=null, pmcid=null, year=2008, volume=1, issue=null, pageStart=117, pageEnd=123, url=http://d.old.wanfangdata.com.cn/NSTLQK/NSTL_QKJJ0231383437/, language=null, rfNumber=[15], rfOrder=14, authorNames=Carl JW, Bai XF, journalName=Int J Clin Exp Pathol, refType=null, unstructuredReference= Carl JW , Bai XF . IL27:its roles in the induction and inhibition of inflammation[J]. Int J Clin Exp Pathol, 2008, 1: 117-123., articleTitle=IL27:its roles in the induction and inhibition of inflammation, refAbstract=null), Reference(id=1222469713314177474, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, doi=10.1158/2326-6066.CIR-16-0022, pmid=null, pmcid=null, year=2016, volume=4, issue=null, pageStart=650, pageEnd=657, url=null, language=null, rfNumber=[16], rfOrder=15, authorNames=McNamara MJ, Hilgart-Martiszus I, Barragan Echenique DM, journalName=Cancer Immunol Res, refType=null, unstructuredReference= McNamara MJ , Hilgart-Martiszus I , Barragan Echenique DM et al . Interferon-γ production by peripheral lymphocytes predicts survival of tumor-bearing mice receiving dual PD-1/CTLA-4 blockade[J]. Cancer Immunol Res, 2016, 4: 650-657., articleTitle=Interferon-γ production by peripheral lymphocytes predicts survival of tumor-bearing mice receiving dual PD-1/CTLA-4 blockade, refAbstract=null), Reference(id=1222469713385480645, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, doi=10.1016/S0065-2776(07)96002-2, pmid=null, pmcid=null, year=2007, volume=96, issue=null, pageStart=41, pageEnd=101, url=null, language=null, rfNumber=[17], rfOrder=16, authorNames=Schoenborn JR, Wilson CB, journalName=Adv Immunol, refType=null, unstructuredReference= Schoenborn JR , Wilson CB . Regulation of interferon-γ during innate and adaptive immune responses[J]. Adv Immunol, 2007, 96: 41-101., articleTitle=Regulation of interferon-γ during innate and adaptive immune responses, refAbstract=null), Reference(id=1222469713460978120, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1222469707249213691, doi=null, pmid=null, pmcid=null, year=2019, volume=14, issue=null, pageStart=127, pageEnd=135, url=null, language=null, rfNumber=[18], rfOrder=17, authorNames=Li L, Lai FF, Du TT, journalName=Chin Med Biotechnol (中国医药生物技术), refType=null, unstructuredReference= Li L , Lai FF , Du TT et al . Effect of different stimulators on PD-1 and TIM3 expression in lymphocytes[J]. 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不同刺激剂对人淋巴细胞活化的影响
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闫征 , 薛妮娜 , 季鸣 , 来芳芳 , 陈晓光 *
药学学报 | 研究论文 2019,54(10): 1863-1867
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药学学报 |研究论文 2019 , 54 (10) : 1863 -1867
不同刺激剂对人淋巴细胞活化的影响
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闫征, 薛妮娜, 季鸣, 来芳芳, 陈晓光*
作者信息
  • 中国医学科学院、北京协和医学院药物研究所, 天然药物活性物质与功能国家重点实验室/创新药物非临床药物代谢及PK/PD研究北京市重点实验室, 北京 100050
通讯作者:
*陈晓光, Tel:86-10-63165207, E-mail:
The effects of different stimulators on the activation of human lymphocytes
Zheng YAN, Ni-na XUE, Ming JI, Fang-fang LAI, Xiao-guang CHEN*
Affiliations
  • State Key Laboratory of Bioactive Substances and Functions of Natural Medicines/Beijing Key Laboratory of Non-clinical Drug Metabolism and PK/PD Study, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China
出版时间: 2019-10-12 doi: 10.16438/j.0513-4870.2019-0670
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本文研究不同刺激剂在体外对人淋巴细胞活化的影响。采用梯度离心法分离获得人外周血单个核细胞(peripheral blood mononuclear cells,PBMC),血液样本由中国食品药品检定研究院提供,并通过伦理委员会批准。用CD3/CD28抗体(anti-CD3/CD28 antibody)、植物血凝素(phytohaemagglutinin,PHA)、金黄色葡萄球菌B型肠毒素(Staphylococcus auresus enterooxin B,SEB)、白介素27(interleukin 27,IL27)、佛波酯(phorbol myristrate acetate,PMA)+离子霉素(ionomycin)在体外分别刺激24 h,观察CD3+CD4+和CD3+CD8+ T细胞亚群CD69表达。同时采用CellTiter-Glo发光法和ELISA法检测淋巴细胞增殖和IFNγ的分泌。CD3/CD28抗体、PMA+ionomycin、SEB和IL27作用PBMC 24 h不影响淋巴细胞增殖,但可以很好地活化CD3+CD4+和CD3+CD8+ T细胞,表现为CD69的平均荧光强度显著右移,并促进淋巴细胞IFNγ的分泌。CD3/CD28抗体、PMA+ionomycin、SEB和IL27是T细胞活化的有效刺激剂。

CD69  /  干扰素γ  /  淋巴细胞增殖  /  淋巴细胞活化

The research aims to study the effects of different stimulants on the activation of human lymphocytes. Human peripheral blood mononuclear cells were prepared by density centrifugation. The blood's sample was provided by National Institutes for Food and Drug Control and approved by its Ethics Committee. The expressions of CD69 in CD3+CD4+ and CD3+CD8+ human T cells were detected by flow cytometry after administrated with CD3/CD28 antibody, phytohaemagglutinin (PHA), Staphylococcus auresus enterooxin B (SEB), interleukin (IL27) and PMA plus ionomycin for 24 h. The proliferation of lymphocyte was detected by CellTiter-Glo kit. The secreted IFNγ in supernatant of medium was examined by ELISA kit. The proliferation of lymphocytes had no change after exposed of CD3/CD28 antibody, SEB, IL27 and PMA plus ionomycin for 24 h. However, the CD69 expressions in CD3+CD4+ and CD3+CD8+ T cells and IFNγ productions were significantly increased by CD3/CD28 antibody, SEB, IL27 and PMA plus ionomycin at 24 h, indicating that CD3+CD4+ and CD3+CD8+ T cells were activated under above-mentioned stimulated condition. CD3/CD28 antibody, SEB, IL27 and PMA plus ionomycin were valid stimulants for T cell activation.

CD69  /  interferon γ  /  lymphocyte proliferation  /  lymphocyte activation
闫征, 薛妮娜, 季鸣, 来芳芳, 陈晓光. 不同刺激剂对人淋巴细胞活化的影响. 药学学报, 2019 , 54 (10) : 1863 -1867 . DOI: 10.16438/j.0513-4870.2019-0670
Zheng YAN, Ni-na XUE, Ming JI, Fang-fang LAI, Xiao-guang CHEN. The effects of different stimulators on the activation of human lymphocytes[J]. Acta Pharmaceutica Sinica, 2019 , 54 (10) : 1863 -1867 . DOI: 10.16438/j.0513-4870.2019-0670
肿瘤免疫疗法已成为近年来肿瘤治疗的研究热点。该治疗手段主要是提高自身免疫系统肿瘤微环境(tumours microenviroment, TME)中肿瘤特异性T细胞(tumour-specific T cell)的免疫应答来杀伤或抑制肿瘤。其中CD3+CD8+ T在TME中起着非常重要的作用, 它属于细胞毒性T淋巴细胞(cytotoxic T lymphocytes, CTLs), 可特异性识别肿瘤细胞表面抗原, 进而靶向杀伤肿瘤细胞[1, 2]。而CD3+CD4+ T归属于辅助性T细胞(T helper cells, Th), 它能通过胞溶机制直接清除肿瘤细胞或间接调节TME发挥抗肿瘤作用。此外, Th辅助细胞可增加CTLs的克隆数量, 提高其分泌IFNγ的能力, 增强CTLs抑瘤活性[1, 3, 4]。因此, 评价CD3+CD4+和CD3+CD8+ T细胞功能在肿瘤免疫治疗分子机制及其药物研发具有重要意义。本实验采用不同的刺激剂CD3/CD28抗体、金黄色葡萄球菌B型肠毒素(Staphylococcus auresus enterooxin B, SEB)、白介素27 (interleukin 27, IL27)、植物血凝素(phytohaemagglutinin, PHA)和佛波酯+离子霉素(PMA+ionomycin)作用于人外周血单个核细胞(peripheral blood mononuclear cells, PBMC) 24 h, 观察CD3+CD4+ T和CD3+CD8+ T细胞的活化标志物CD69的变化以及淋巴细胞增殖和IFNγ分泌能力。一方面综合评价这些生物学指标能否反映T功能变化; 另一方面对比这些刺激剂活化T细胞的强度, 从而为后续肿瘤免疫研究和药物筛选提供实验依据。
血液样本  新鲜健康人全血, 由中国食品药品检定研究院提供, 已取得志愿者的知情同意, 并得到中国食品药品检定研究院伦理委员会批准。
试剂  人外周血淋巴细胞分离液购于达科为生物技术有限公司。CellTiter-Glo检测试剂盒购于Promega公司, IL27、PHA、SEB、PMA、ionomycin购于Sigma公司。IFNγ的ELISA试剂盒、CD3/CD28抗体和Pecy7 anti-human CD69抗体购自Biolegend公司。
仪器  美国BD公司的FACSVerse流式细胞仪, 日本三洋公司的CO2培养箱, 美国Sigma公司冷冻离心机, 美国Bio-tec酶标仪。
人PBMC的制备  取新鲜抗凝人全血至无菌的50 mL离心管, 再用等体积的磷酸盐缓冲液(phosphate buffer saline, PBS)稀释全血。在另一离心管中加入一定体积的分离液, 将稀释后的全血小心缓慢地平铺到分离液面上方, 使两界面保持清晰。淋巴细胞分离液、未稀释全血、PBS体积比为1:1:1。于室温并置于水平转子离心机, 800 ×g离心25 min。离心结束后, 小心吸取离心管中部致密白色细胞层(PBMC层)移至新的无菌离心管中。用PBS稀释到一定体积, 颠倒混匀。于室温水平转子250 ×g离心10 min, 弃上清。重复洗涤2次, 进行细胞计数后调整细胞数至1×106个/孔。
流式细胞术检测不同刺激剂对淋巴细胞的活化作用  将制备好的PBMC接种于96孔U型板中, 置于37 ℃、5% CO2的细胞培养箱中平衡2 h。加入不同刺激剂, 如抗人CD3/CD28抗体(终质量浓度: 20和200 ng·mL-1)、SEB (终质量浓度10 ng·mL-1)、PHA (终质量浓度1 μg·mL-1)、IL27 (终质量浓度10 ng·mL-1)、PMA + ionomycin联合刺激(终质量浓度为PMA: 10 ng·mL-1、ionomycin: 1 μg·mL-1)。每个剂量组设3个平行孔, 置于37 ℃、5% CO2的细胞培养箱中继续培养24 h。离心, 收集细胞上清, 冻于-80 ℃冰箱保存。将细胞用预冷PBS洗涤2次, 加入Pecy7 anti-human CD69抗体孵育30 min, 预冷的PBS洗涤2次后, 采用流式细胞检测仪(BD FACS Verse)检测淋巴细胞CD69的表达情况, 并采用Flowjo 7.6软件分析淋巴细胞CD69的平均荧光强度和CD69阳性淋巴细胞百分比。
CellTiter-Glo发光法检测淋巴细胞增殖  将上述处理的细胞重悬, 移取100 μL细胞悬液至新的96孔板中, 加入100 μL CellTiter-Glo底物, 室温避光5 min, 于Bio-tec酶标仪检测发光值。
ELISA试剂盒检测IFNγ  将试剂盒中Capture antibody加入96孔板中, 4 ℃过夜孵育。次日用清洗缓冲液清洗5次, 向每孔中加入待测样品及不同浓度的标准品于室温孵育2 h, 用清洗缓冲液洗5次。加入检测抗体于室温孵育1 h, 清洗缓冲液洗5次。加入辣根过氧化物酶(HRP)标记的二抗室温孵育30 min。再加入底物TMB于室温避光孵育15 min, 加入硫酸终止, 在酶标仪450 nm处检测。
统计学方法  采用GraphPad Prism统计软件处理, 数据用x ± s表示。P≤0.05为具有统计学意义。
采用流式细胞术, 对CD3+CD4+ T细胞亚群中的CD69表达进行检测。如图 1所示, 静息状态下CD3+CD4+ T细胞表面上的CD69表达量较低。CD3/CD28 (20和200 ng·mL-1)、PMA (10 ng·mL-1)+ionomycin (1 μg·mL-1)、SEB (10 ng·mL-1)和IL27 (10 ng·mL-1)均可以显著性促进CD69的表达, 表现为PE-Cy7通道的平均荧光强度右移。其中PMA+ionomycin诱导CD3+CD4+ T细胞上CD69的表达能力最强, CD3/CD28、SEB和IL27刺激因子诱导CD3+CD4+ T细胞上CD69的表达能力基本一致。上述这些因子诱导CD69高表达的CD3+CD4+ T细胞百分比分别为74.7%、91.5%、85.2%、64.1%、68.3%。而PHA (1 μg·mL-1)对CD3+CD4+ T细胞上CD69的表达无诱导作用, CD3+CD4+CD69+ T细胞百分比基本无影响。
本研究也检测了不同刺激因子对CD3+CD8+ T细胞亚群中的CD69表达的影响。如图 2所示, 与CD3+CD4+ T细胞一致, 在未刺激状态下CD3+CD8+ T细胞表面上的CD69表达量较低。CD3/CD28 (20和200 ng·mL-1)、PMA (10 ng·mL-1)+ionomycin (1 μg·mL-1)、SEB (10 ng·mL-1)、IL27 (10 ng·mL-1)和PHA (1 μg·mL-1)均可以促进CD3+CD8+ T细胞CD69的表达。其中CD3/CD28、PMA+ionomycin、SEB和IL27促进CD3+CD8+ T细胞CD69的表达能力基本一致, 都显著强于PHA。上述这些因子诱导CD69高表达的CD3+CD8+ T细胞百分比分别为88.3%、92.4%、88.5%、70.6%、82.9%、15.1%。
T细胞增殖对机体的免疫应答起到了重要作用。本研究采用CellTiter-Glo®发光法检测了不同刺激剂对淋巴细胞增殖能力的检测。如图 3所示, CD3/CD28、PMA+ionomycin、SEB、IL27和PHA作用24 h均无明显促进人淋巴细胞增殖的作用。
IFNγ是T淋巴细胞活化的重要标志。采用ELISA法检测了不同刺激剂对淋巴细胞分泌IFNγ含量的影响。如图 4所示, CD3/CD28 (200 ng·mL-1)、PMA (10 ng·mL-1)+ionomycin (1 μg·mL-1)、SEB (10 ng·mL-1)、IL27 (10 ng·mL-1)作用24 h显著诱导由淋巴细胞分泌的IFNγ。其中, PMA+ionomycin诱导IFNγ分泌的作用最强。
T细胞的功能, 主要从T细胞增殖、活化和分泌水平方面进行评价。增殖能力检测常用形态学观察、放射性核素掺入法和MTT法。镜下形态观察效率低且主观影响大; 同位素虽然精确, 但会对环境造成污染; MTT法灵敏度不够, 同时会受到化合物自身的颜色干扰。本研究采用CellTiter-Glo发光法检测淋巴细胞的增殖。该方法通过检测ATP含量, 可快速灵敏地反映活细胞数量, 成为目前免疫细胞增殖检测的一种常用方法[5, 6]。活化能力评价的方法有CFSE标记检测细胞分裂[7], ELISA法检测细胞因子分泌如IFNγ, 流式细胞术检测细胞表面标志物CD69、CD25和HLA-DR等[8-10]。本研究则检测不同功能刺激剂对T细胞亚群细胞表面标志物CD69表达和IFNγ分泌。其中, CD3/CD28抗体为体外模拟特异性T淋巴细胞活化信号而诱导T细胞的增殖[11]; PMA激活PKC信号从而使T细胞活化增殖[12], ionomycin为Ca2+载体并介导Ca2+的内流[13], 与PMA联用进一步增强其激活淋巴细胞的能力; SEB为超级抗原, 通过T细胞受体β (TCR-β)大量激活T细胞[14]; PHA是一种有丝分裂原从而激活淋巴细胞[7]; IL27与其受体相互作用诱导早期Th1型反应[15]。在本实验所用浓度下的CD3/CD28抗体、PMA+ionomycin、SEB和IL27作用刺激24 h, CD3+CD4+和CD3+CD8+ T细胞CD69的表达显著增高。以上结果提示, CD69是一个很好的T细胞早期活化的标志物, 可同时反映CD3+CD4+和CD3+CD8+ T的活化。与此同时, 淋巴细胞分泌的IFNγ含量也显著增高。IFNγ是T淋巴细胞活化的重要指征之一[16]。IFNγ最初由机体固有免疫中自然杀伤细胞(NK)和自然杀伤T (NKT)细胞分泌, 然后被特异性抗原免疫中Th1和CTLs细胞进一步分泌[17]。因此, 应该综合活化标志物CD69与IFNγ含量来评价T细胞的活化状态。而刺激剂作用24 h对淋巴细胞增殖几乎无影响, 提示淋巴细胞在接受刺激后短时间内以活化为主, 随着时间的延长才可能表现出增殖能力增强。
由于小分子化合物体外单独作用不能直接刺激淋巴细胞活化, 因此在体外评价小分子化合物对T细胞功能的影响时, 需要模拟肿瘤微环境的抗原呈递细胞(APCs)与T细胞的相互作用, 或者采用刺激因子先将T细胞活化[18]。本研究除了发现反映T细胞活化的标志物外, 更好地建立了体外活化T细胞的方法, 为进一步抗肿瘤免疫小分子药物发现奠定实验基础。
  • 重大新药创制专项(2018ZX09711001-003)
参考文献 引证文献
排序方式:
[1]
Borst J , Ahrends T , Bąbała N et al . CD4+ T cell help in cancer immunology and immunotherapy[J]. Nat Rev Immunol, 2018, 18: 635-647.
[2]
Galluzzi L , Vacchelli E , Bravo-San Pedro JM et al . Classification of current anticancer immunotherapies[J]. Oncotarget, 2014, 5: 12472-12508. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4350348/
[3]
Melssen M , Slingluff CL Jr . Vaccines targeting helper T cells for cancer immunotherapy[J]. Curr Opin Immunol, 2017, 47: 85-92.
[4]
Bevan MJ . Helping the CD8+ T-cell response[J]. Nat Rev Immunol, 2004, 4: 595-602.
[5]
Mollgard L , Tidefelt U , Sundman-Engberg B et al . In vitro chemosensitivity testing in acute non lymphocytic leukemia using the bioluminescence ATP assay[J]. Leuk Res, 2000, 24: 445-452.
[6]
Ishizaka A , Tono-oka T , Matsumoto S . Evaluation of the proliferative response of lymphocytes by measurement of intracellular ATP[J]. J Immunol Methods, 1984, 72: 127-132.
[7]
Xue NN , Dong K , Lai FF et al . CFSE-labeled proliferative assays for assessment of T cell function induced by different stimulants[J]. J Harbin Univ Commerce (Nat Sci Ed) (哈尔滨商业大学学报(自然科学版)), 2017, 33: 129-134. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=hljsxyxb201702001
[8]
Rea IM , McNerlan SE , Alexander HD . CD69, CD25, and HLA-DR activation antigen expression on CD3+ lymphocytes and relationship to serum TNF-α, IFN-γ, and sIL-2R levels in aging[J]. Exp Gerontol, 1999, 34: 79-93.
[9]
Theze J , Alzari PM , Bertoglio J . Interleukin 2 and its receptors:recent advances and new immunological functions[J]. Immunol Today, 1996, 17: 481-486.
[10]
Ziegler SF , Ramsdell F , Alderson MR . The activation antigen CD69[J]. Stem Cells, 1994, 12: 456-465.
[11]
Trickett A , Kwan YL . T cell stimulation and expansion using anti-CD3/CD28 beads[J]. J Immunol Methods, 2003, 275: 251-255.
[12]
Seo HH , Lee CY , Lee J et al . The role of nuclear factor of activated T cells during phorbol myristate acetate-induced cardiac differentiation of mesenchymal stem cells[J]. Stem Cell Res Ther, 2016, 7: 90.
[13]
Nakamura S , Nakanishi A , Takazawa M et al . Ionomycin-induced calcium influx induces neurite degeneration in mouse neuroblastoma cells:analysis of a time-lapse live cell imaging system[J]. Free Radic Res, 2016, 50: 1214-1225.
[14]
Xu SX , Kasper KJ , Zeppa JJ et al . Superantigens modulate bacterial density during Staphylococcus aureus nasal colonization[J]. Toxins (Basel), 2015, 7: 1821-1836.
[15]
Carl JW , Bai XF . IL27:its roles in the induction and inhibition of inflammation[J]. Int J Clin Exp Pathol, 2008, 1: 117-123. http://d.old.wanfangdata.com.cn/NSTLQK/NSTL_QKJJ0231383437/
[16]
McNamara MJ , Hilgart-Martiszus I , Barragan Echenique DM et al . Interferon-γ production by peripheral lymphocytes predicts survival of tumor-bearing mice receiving dual PD-1/CTLA-4 blockade[J]. Cancer Immunol Res, 2016, 4: 650-657.
[17]
Schoenborn JR , Wilson CB . Regulation of interferon-γ during innate and adaptive immune responses[J]. Adv Immunol, 2007, 96: 41-101.
[18]
Li L , Lai FF , Du TT et al . Effect of different stimulators on PD-1 and TIM3 expression in lymphocytes[J]. Chin Med Biotechnol (中国医药生物技术), 2019, 14: 127-135.
2019年第54卷第10期
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doi: 10.16438/j.0513-4870.2019-0670
  • 接收时间:2019-08-22
  • 首发时间:2026-01-26
  • 出版时间:2019-10-12
补充材料
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出版历史
  • 收稿日期:2019-08-22
  • 修回日期:2019-09-06
基金
重大新药创制专项(2018ZX09711001-003)
作者信息
    中国医学科学院、北京协和医学院药物研究所, 天然药物活性物质与功能国家重点实验室/创新药物非临床药物代谢及PK/PD研究北京市重点实验室, 北京 100050

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*陈晓光, Tel:86-10-63165207, E-mail:
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https://castjournals.cast.org.cn/joweb/yxxb/CN/10.16438/j.0513-4870.2019-0670
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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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