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Simian fetal brain progenitor cells for studying viral neuropathogenesis

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Abstract

The pathogenesis of neurologic dysfunctions caused by human immunodeficiency virus type 1 (HIV-1) infection is not yet well understood. Simian immunodeficiency virus (SIV) infection of macaques is an important animal model for HIV-1 infection. This is the first report to characterize brain progenitor cells (BPCs) isolated from embryonic brain of cynomolgus monkeys (Macaca fascicularis) by neurosphere assay and utilize BPC-derived cell culture for studying SIV infection. The self-renewal and multilineage differentiation properties of BPCs are convenient for planning viral infection experiments. The BPC-derived culture does not contain macrophage/microglial cells, fibroblasts, or endothelial cells. Thus, this culture is appropriate for studying direct relation between SIV infection and neuronal and glial cells. First, the authors characterized undifferentiated and differentiated simian BPCs by immunocytochemistry, flow cytometry analysis, real-time polymerase chain reaction (PCR), and reverse transcriptase (RT)-PCR. The BPCs induced to differentiate by the addition of 1% fetal bovine serum (FBS) were composed of heterogeneous cells expressing nestin, glial fibrillary acidic protein (GFAP), and/or tubulin beta III isoform (Tuj). None of them expressed the monocyte/macrophage/microglial marker. mRNA expression of CD4, CXCR4, CCR5, GPR1, STRL33, and APJ in both undifferentiated and differentiated BPCs were shown by RT-PCR method, suggesting that SIV would infect and replicate in this culture system. Then, it was confirmed that the neurotropic SIV strain, SIV17/E-Fr, replicated productively in BPC-derived cells. The SIV/17E-FrΔnefGFP was inoculated to identify the infected cells and immunocytochemistry analysis revealed that green fluorescent protein (GFP)-expressing cells were mostly GFAP positive and coexpressed with SIV p27 antigen. Thus, BPC-derived cell culture system is applicable for studying SIV infection in glial and neuronal cells.

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Authors and Affiliations

  1. Department of Pathology, National Institute of Infectious Diseases, Tokyo, Japan

    Naoko Iwata, Hiroaki Yoshida, Minoru Tobiume, Tetsutaro Sata & Noriko Nakajima

  2. Department of Applied Biochemistry, Tokai University, Kanagawa, Japan

    Hiroaki Yoshida

  3. Tsukuba Primate Center, National Institute of Biomedical Innovation, Tokyo, Japan

    Fumiko Ono

  4. Department of Physiology, Keio University School of Medicine, Tokyo, Japan

    Takuya Shimazaki

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  1. Naoko Iwata
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  2. Hiroaki Yoshida
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  3. Minoru Tobiume
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  4. Fumiko Ono
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  5. Takuya Shimazaki
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  6. Tetsutaro Sata
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  7. Noriko Nakajima
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Correspondence to Noriko Nakajima.

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This work was supported by a Grant-in-Aid from the Ministry of Health, Labour and Welfare for AIDS research (16100601).

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Iwata, N., Yoshida, H., Tobiume, M. et al. Simian fetal brain progenitor cells for studying viral neuropathogenesis. Journal of NeuroVirology 13, 11–22 (2007). https://doi.org/10.1080/13550280601086064

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