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张立, 贲昆龙. 2001: 树qu免疫细胞体外感染Ⅰ型人免疫缺陷病毒的实验研究. 动物学研究, 22(1): 33-40.
引用本文: 张立, 贲昆龙. 2001: 树qu免疫细胞体外感染Ⅰ型人免疫缺陷病毒的实验研究. 动物学研究, 22(1): 33-40.
ZHANG Li, BEN Kun-Long. 2001. In vitro Infection of Tree Shrew Immunocytes with Human Immunodeficiency Virus Type Ⅰ. Zoological Research, 22(1): 33-40.
Citation: ZHANG Li, BEN Kun-Long. 2001. In vitro Infection of Tree Shrew Immunocytes with Human Immunodeficiency Virus Type Ⅰ. Zoological Research, 22(1): 33-40.

树qu免疫细胞体外感染Ⅰ型人免疫缺陷病毒的实验研究

In vitro Infection of Tree Shrew Immunocytes with Human Immunodeficiency Virus Type Ⅰ

  • 摘要: 至今可以感染Ⅰ型人免疫缺陷病毒(HIV-1)的动物只有黑猩猩和长臂猿,这严重阻碍了HIV-1的疫苗研究和治疗研究。因此,寻找新的可以感染HIV-1的动物模型成为十分迫切的课题。已知树qu对许多重要的医学病毒易感,为了探讨树qu是否可以感染HIV-1,利用不同辅助受体的5种HIV-1病毒株,体外感染云南野生成年树qu的淋巴细胞和单核/巨噬细胞;同时还用这些病毒感染人外周血淋巴细胞或单核细胞。然后用RT-PCR、PCR和流式细胞术分别进行检测。用RT-PCR方法未检测到感染上清中有病毒粒子的存在,用PCR法未能发现树qu的这些免疫细胞中有前病毒DNA,用流式细胞术也未能在这些感染HIV-1的树qu细胞的表面检测到特异抗原;而感染HIV-1的人免疫细胞均为阳性结果。实验结果表明树qu的这些免疫细胞在体外未能感染上HIV-1,可能的原因是树qu的这些免疫细胞的HIV-1受体(CD4)和辅助受体(CCR5或CXCR4)与人的免疫细胞差别较大。

     

    Abstract: The acquired immunodeficiency syndrome caused by HIV-1 is spreading all over the world.The slow progress in AIDS therapy and vaccine partially imputes to be lack of appropriate animal models used to study AIDS pathogenesis and to evaluate vaccines.To find an animal model is an imperative task in HIV research.Tree shrew is widely used in biomedical research,and is susceptible to many medically important viruses.Whether tree shrew (Tupaia belangeri) can be infected by HIV-1 is a valuable approach.In present study,wild tree shrews from Yunnan Province were maintained in laboratory animal facility for more than 2 weeks before sacrifice.Tree shrew spleen and peripheral blood lymphocytes and monocytes/macrophages and human peripheral blood lymphocytes and monocytes were separately infected by 5 HIV-1 strains (HIV-1ⅢB,HIV-1JR-FL,HIV-1Ada-M,HIV-1Ba-L,HIV-1SF162),which use different coreceptors after the cells were activated by allogeneic lymphocytes,PHA and IL-2 for 72 h.The HIV-1 infected cells were then cultured in vitro for 15 days.The proliferation rates and viability of HIV-1 infected human immunocytes obviously dropped on day 15,but those of tree shrew immunocytes did not despite either infected or not infected by HIV-1.HIV-1 particles in the infected culture supernatant and proviral DNA in the cells were respectively detected by RT-PCR using primer sk145/431 and by PCR using primer sk68/69 on days 1,3,5,7,9,12,15 after infection with HIV-1.In addition,the expression of HIV-1 specific antigens on the tree shrew and human immunocytes after infection were detected by flow cytometry,for which an AIDS patient plasma was used as the first antibody,and FITC-conjugated sheep anti-human-IgG as the second antibody.HIV-1 RNA and proviral DNA were respectively found in the culture supernatant of the HIV-1 infected human immunocytes and in the infected cells.Using flow cytometry,HIV-1 specific antigens were also measurable on the surface of these cells.However,neither HIV-1 RNA in the supernatant from the infected HIV-1 tree shrew immunocytes nor the proviral DNA from the infected tree shrew immunocytes could be detected.The HIV-1 specific antigens were not demonstrated on the surface of the HIV-1 infected tree shrew immunocytes.Taken together,these experimental results suggested that the tree shrew immunocytes can not be in vitro infected by these HIV-1 strains.It seems to be caused by the structure differences of the HIV-1 receptors (CD4) and coreceptors (CCR5 or CXCR4) between human and tree shrew immunocytes.

     

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