贵州医科大学学报

2011, v.36;No.148(01) 5-8

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登革2型病毒粘附人血小板的体外研究初探
A Preliminary Study on Adherence of Type 2 Dengue Virus to Human Platelets in Vitro

张敏;商正玲;左丽;
ZHANG Min,SHANG Zhengling,ZUO Li(Department of Immunology,Guiyang Medical College,Guiyang 550004,Guizhou,China)

摘要(Abstract):

目的:了解登革2型病毒(DEN-2)是否能在体外粘附人血小板并制备抗人血小板抗体。方法:密度梯度离心法分离血小板,DEN-2与血小板体外共孵育2 h,间接免疫荧光法分析DEN-2粘附人血小板情况。采用TritonX-100裂解法制备血小板蛋白并经皮下注射免疫BALB/C小鼠获取抗血小板血清,经Western blot、间接免疫荧光及间接ELISA对抗血清进行鉴定及效价测定。结果:间接免疫荧光法提示DEN-2可体外粘附人血小板;Western blot、间接免疫荧光法表明获得特异性抗人血小板抗体,间接ELISA测定抗体效价可达到1∶12 800。结论:DEN-2能够在体外粘附人血小板,血小板蛋白免疫BALB/C小鼠后可产生较高效价的抗人血小板抗体,为今后研究血小板在DEN-2感染中的免疫学功能奠定一定的基础。
Objective:To investigate whether type 2 dengue virus(DEN-2) could adhere to human platelets(PLT) in vitro and to prepare mouse anti-human platelet antibody.Methods: Density gradient centrifugation was used to isolate PLT.DEN-2 was co-cultured with human PLT for 2 hours.DEN-2 adhering to PLT was detected with indirect immunofluorescence assay(IFA).The antiserum against PLT was prepared by subcutaneous injection of PLT proteins extracted with Triton X-100 into BALB/C rats.The specificity of anti-PLT antibodies was identified by Western blot and IFA.Titration of them was examined with indirect ELISA.Results: DEN-2 could adhere to human PLT.The specific anti-human PLT antibodies were obtained by Western blot and identified by IFA.The titration of the antibodies was 1∶12 800 detected by indirect ELISA.Conclusions: DEN-2 could adhere to human PLT in vitro.Specific anti-human PLT antibodies with high titration could be prepared by immunizing BALB/C mice with PLT proteins.These results lay a foundation for further study of PLT immune function in DEN-2 caused infectious diseases.

关键词(KeyWords): 登革热病毒;血小板;抗体;荧光免疫测定;小鼠,近交BALBC
dengue virus;blood platelets;antibodies;fluoroimmunoassay;mice,inbred BALBC

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基金项目(Foundation): 国家自然科学基金项目(31060129)

作者(Author): 张敏;商正玲;左丽;
ZHANG Min,SHANG Zhengling,ZUO Li(Department of Immunology,Guiyang Medical College,Guiyang 550004,Guizhou,China)

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DOI: 10.19367/j.cnki.1000-2707.2011.01.003

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