Next-Generation mRNA Sequencing Reveals Pyroptosis-Induced CD4+ T Cell Death in Early Simian Immunodeficiency Virus-Infected Lymphoid Tissues
Next-Generation mRNA Sequencing Reveals Pyroptosis-Induced CD4+ T Cell Death in Early Simian Immunodeficiency Virus-Infected Lymphoid Tissues
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DOI：
10.1128/jvi.02297-15
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发表时间：
2016-01-01
影响因子：
5.4
通讯作者：
Li, Qingsheng
中科院分区：
文献类型：
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作者：
Lu, Wuxun;Demers, Andrew J.;Li, Qingsheng
淋巴组织 (LT) 是人类免疫缺陷病毒 1 型 (HIV-1) 复制和病毒-宿主相互作用发生的主要场所，导致炎症、免疫激活和 CD4(+) T 细胞死亡等形式的免疫病理学。 HIV-1 在 LT 中的发病机制已得到广泛研究；然而，我们对感染早期阶段病毒与宿主相互作用的了解仍然不完整。我们研究了在直肠内接种猿猴免疫缺陷病毒（SIV）后不同时间（接种后天数[dpi]）的恒河猴直肠引流淋巴结（dLN）中病毒与宿主的相互作用。在 3 dpi 时，使用下一代 mRNA 测序 (RNA-seq) 检测到 103 个差异表达基因 (DEG)。在 6 dpi 和 10 dpi 时，随着 SIV 复制的增加，分别检测到 366 和 1,350 个 DEG，包括编码在先天抗病毒免疫反应、炎症和免疫激活中发挥作用的蛋白质的基因上调。值得注意的是，经典细胞焦亡途径中的基因（IFI16、caspase-1 和白细胞介素 1(+) [IL-1(+)]）的表达显着上调。我们使用流式细胞术进一步验证了焦亡的增加，发现表达活化的 caspase-1 蛋白（持续焦亡的标志）的 CD4(+) T 细胞数量显着增加，这与 dLN 中 CD4(+) T 细胞的减少相关。我们的研究结果表明，在早期SIV感染中，焦亡有助于体内CD4(+) T细胞死亡，这表明焦亡可能在SIV乃至HIV-1的发病机制中发挥关键作用，因为焦亡不仅诱导CD4(+) T细胞死亡，而且放大炎症和免疫激活。因此，阻断 CD4(+) T 细胞焦亡可能是抗逆转录病毒治疗的补充治疗。 重要性 尽管次级淋巴组织 (LT) 是人类免疫缺陷病毒 1 型 (HIV-1) 复制、炎症、免疫激活和 CD4(+) T 细胞死亡的主要部位，但早期感染期间 LT 的免疫发病机制仍然很大程度上未知。使用艾滋病毒直肠感染的猿猴免疫缺陷病毒（SIV）/恒河猴模型，我们研究了早期病毒与宿主的相互作用。我们的结果显示，LTs 早期感染时宿主反应增强，包括抗病毒免疫反应、炎症和免疫激活相关基因的上调。重要的是，参与典型细胞焦亡途径的基因显着上调，并且CD4(+) T细胞减少与表达活化的caspase-1蛋白的CD4(+) T细胞数量增加之间存在很强的相关性，这表明在极早期SIV感染中细胞焦亡导致体内CD4(+) T细胞死亡。我们的研究结果表明，阻断焦亡可能能够减少早期 SIV 感染期间 CD4(+) T 细胞的损失。
Lymphoid tissues (LTs) are the principal sites where human immunodeficiency virus type 1 (HIV-1) replicates and virus-host interactions take place, resulting in immunopathology in the form of inflammation, immune activation, and CD4(+) T cell death. The HIV-1 pathogenesis in LTs has been extensively studied; however, our understanding of the virus-host interactions in the very early stages of infection remains incomplete. We investigated virus-host interactions in the rectal draining lymph nodes (dLNs) of rhesus macaques at different times after intrarectal inoculation (days postinoculation [dpi]) with simian immunodeficiency virus (SIV). At 3 dpi, 103 differentially expressed genes (DEGs) were detected using next-generation mRNA sequencing (RNA-seq). At 6 and 10 dpi, concomitant with increased SIV replication, 366 and 1,350 DEGs were detected, respectively, including upregulation of genes encoding proteins that play a role in innate antiviral immune responses, inflammation, and immune activation. Notably, genes (IFI16, caspase-1, and interleukin 1(+) [IL-1(+)]) in the canonical pyroptosis pathway were significantly upregulated in expression. We further validated increased pyroptosis using flow cytometry and found that the number of CD4(+) T cells expressing activated caspase-1 protein, the hallmark of ongoing pyroptosis, were significantly increased, which is correlated with decreased CD4(+) T cells in dLNs. Our results demonstrated that pyroptosis contributes to the CD4(+) T cell death in vivo in early SIV infection, which suggests that pyroptosis may play a pivotal role in the pathogenesis of SIV, and by extension, that of HIV-1, since pyroptosis not only induces CD4(+) T cell death but also amplifies inflammation and immune activation. Thus, blocking CD4(+) T cell pyroptosis could be a complementary treatment to antiretroviral therapy.IMPORTANCEAlthough secondary lymphoid tissues (LTs) are principal sites of human immunodeficiency virus type 1 (HIV-1) replication, inflammation, immune activation, and CD4(+) T cell death, immunopathogenesis in LTs during early infection remains largely unknown. Using the simian immunodeficiency virus (SIV)/rhesus monkey model of HIV rectal infection, we investigated early virus-host interactions. Our results revealed elevated potent host responses in early infection in LTs, including upregulation of genes involved in antiviral immune response, inflammation, and immune activation. Importantly, genes involved in the canonical pyroptosis pathway were significantly upregulated, and there was a strong correlation between CD4(+) T cell decrease and increased number of CD4(+) T cells expressing activated caspase-1 protein, demonstrating that pyroptosis contributes to CD4(+) T cell death in vivo in very early SIV infection. Our finding suggests that blocking pyroptosis may be able to decrease CD4(+) T cell loss during early SIV infection.