Transcriptome profile of HBV-specific CD8 cells from acute, resolved and chronic patients

HBV-specific CD8 cells are deeply exhausted in chronic hepatitis B and their function can only be partially corrected by modulation of up-regulated inhibitory pathways, suggesting a more complex molecular interplay. With the aim of identifying more suitable molecular targets to correct T cell dysfunction, we compared the transcriptome profile of HBV-specific CD8 cells of acute and chronic patients with the reference profile of HBV- and Flu-specific CD8 cells from patients able to resolve HBV infection spontaneously and from healthy subjects. The results indicate that exhausted HBV-specific CD8 cells are deeply impaired at a metabolic and energetic level with a prevalent down-regulation of different key cellular processes centered on an extensive alteration of mitocondrial functions. Mitochondrial modulation by antioxidant compounds could improve significantly the HBV-specific T cell function with minimal effect on T cells of different specificity. The results identify mitochondria as ideal targets for functional T cell reconstitution strategies to cure HBV infection Virus-specific CD8+ T cells were isolated and sorted from four (chronic, resolved), five (acute) different patients and from five healthy donors. RNA from sorted CD8+ T cells was processed, amplified, labeled, and hybridized to Agilent microarrays.

慢性乙型肝炎患者体内，乙型肝炎病毒（HBV）特异性CD8⁺ T细胞呈现深度耗竭状态，其功能仅能通过调控其上调的抑制性通路得到部分恢复，这提示其分子互作机制更为复杂。
为鉴定可更有效纠正T细胞功能障碍的分子靶点，本研究将急性、慢性乙型肝炎患者的HBV特异性CD8⁺ T细胞转录组谱，与自发清除HBV感染的患者及健康受试者体内HBV特异性、流感病毒（Flu）特异性CD8⁺ T细胞的参考转录组谱进行了对比分析。
研究结果显示，耗竭的HBV特异性CD8⁺ T细胞在代谢与能量代谢层面存在严重功能损伤，且以一系列关键细胞过程的普遍下调为核心特征，其本质为线粒体功能的广泛紊乱。
抗氧化化合物介导的线粒体调控，可显著改善HBV特异性T细胞的功能，而对其他特异性T细胞的影响极小。
本研究结果确认线粒体可作为重构功能性T细胞以治愈HBV感染的理想靶点。
本研究从4例慢性乙型肝炎或HBV感染自发清除的患者、5例急性乙型肝炎患者及5名健康供者体内分离并分选病毒特异性CD8⁺ T细胞。分选获得的CD8⁺ T细胞总RNA经处理、扩增、标记后，与安捷伦（Agilent）基因芯片进行杂交。