Anti-tat Hutat2:Fc mediated protection against tat-induced neurotoxicity and HIV-1 replication in human monocyte-derived macrophages
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  • 作者：Wen Kang (1) (2)
    Wayne A Marasco (3)
    Hsin-I Tong (2)
    Mary Margaret Byron (4)
    Chengxiang Wu (2)
    Yingli Shi (2)
    Si Sun (2)
    Yongtao Sun (1)
    Yuanan Lu (2)

    1. Department of Infectious Diseases     ; Tangdu Hospital ; The Fourth Military Medical University ; 569 Xinsi Road ; Xi鈥檃n ; Shaanxi ; 710038 ; China
    2. Department of Public Health Sciences     ; John A. Burns School of Medicine ; University of Hawaii ; 1960 East鈥搘est Road ; Honolulu ; HI ; 96822 ; USA
    3. Department of Cancer Immunology and AIDS     ; Dana-Farber Cancer Institute ; Harvard Medical School ; 50 Brookline Avenue ; Boston ; MA ; 02215 ; USA
    4. Hawaii Center for AIDS     ; John A. Burns School of Medicine ; University of Hawaii ; 651 Ilalo St. ; BSB ; Suite 231 ; Honolulu ; HI ; 96813 ; USA
  • 关键词：Anti ; Tat antibody ; HIV ; 1 ; HIV ; associated neurocognitive disorders ; Human monocyte ; derived macrophages ; Lentivirus ; Neuroprotection
  • 刊名：Journal of Neuroinflammation
  • 出版年：2014
  • 出版时间：December 2014
  • 年：2014
  • 卷：11
  • 期：1
  • 全文大小：2,237 KB
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  • 刊物主题：Neurosciences; Neurology; Neurobiology; Immunology;
  • 出版者：BioMed Central
  • ISSN：1742-2094
文摘
Background HIV-1 Tat is essential for HIV replication and is also a well-known neurotoxic factor causing HIV-associated neurocognitive disorder (HAND). Currently, combined antiretroviral therapy targeting HIV reverse transcriptase or protease cannot prevent the production of early viral proteins, especially Tat, once HIV infection has been established. HIV-infected macrophages and glial cells in the brain still release Tat into the extracellular space where it can exert direct and indirect neurotoxicity. Therefore, stable production of anti-Tat antibodies in the brain would neutralize HIV-1 Tat and thus provide an effective approach to protect neurons. Methods We constructed a humanized anti-Tat Hutat2:Fc fusion protein with the goal of antagonizing HIV-1 Tat and delivered the gene into cell lines and primary human monocyte-derived macrophages (hMDM) by an HIV-based lentiviral vector. The function of the anti-Tat Hutat2:Fc fusion protein and the potential side effects of lentiviral vector-mediated gene transfer were evaluated in vitro. Results Our study demonstrated that HIV-1-based lentiviral vector-mediated gene transduction resulted in a high-level, stable expression of anti-HIV-1 Tat Hutat2:Fc in human neuronal and monocytic cell lines, as well as in primary hMDM. Hutat2:Fc was detectable in both cells and supernatants and continued to accumulate to high levels within the supernatant. Hutat2:Fc protected mouse cortical neurons against HIV-1 Tat86-induced neurotoxicity. In addition, both secreted Hutat2:Fc and transduced hMDM led to reducing HIV-1BaL viral replication in human macrophages. Moreover, lentiviral vector-based gene introduction did not result in any significant changes in cytomorphology and cell viability. Although the expression of IL8, STAT1, and IDO1 genes was up-regulated in transduced hMDM, such alternation in gene expression did not affect the neuroprotective effect of Hutat2:Fc. Conclusions Our study demonstrated that lentivirus-mediated gene transfer could efficiently deliver the Hutat2:Fc gene into primary hMDM and does not lead to any significant changes in hMDM immune-activation. The neuroprotective and HIV-1 suppressive effects produced by Hutat2:Fc were comparable to that of a full-length anti-Tat antibody. This study provides the foundation and insights for future research on the potential use of Hutat2:Fc as a novel gene therapy approach for HAND through utilizing monocytes/macrophages, which naturally cross the blood-brain barrier, for gene delivery.

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