Department of Laboratory Medicine, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, China.
Department of Laboratory Medicine, Guizhou Provincial People's Hospital, Guiyang, China.
BMC Biotechnol. 2024 Oct 14;24(1):80. doi: 10.1186/s12896-024-00908-8.
Hepatitis B virus (HBV) clearance depends on an effective adaptive immune response, especially HBV-specific T cell-mediated cellular immunity; however, it is difficult to produce enough HBV-specific T cells effectively.
In this work, we investigated the proportions of stimulated cells, serum, and culture media as the three primary factors to determine the most effective procedure and applied it to HLA-A2 (+) people. In parallel, we also examined the correlation between clinical parameters and HBV-specific immunity. Concerning amplification efficiency, 4 × 10 cells stimulation was superior to 2 × 10 cells stimulation, AIM-V medium outperformed 1640 medium, and fetal bovine serum (FBS) exceeded human AB serum under comparable conditions. As expected, this procedure is also suitable for developing HBV-specific CD8 + T cells in HLA-A2(+) individuals. Expanded HBV-specific T cell responses decreased with treatment time and were negatively correlated with HBV DNA and HBsAg. Furthermore, the number of HBV-specific IFN-γ + SFCs was strongly correlated with the ALT level and negatively correlated with the absolute lymphocyte count and the ALB concentration.
We confirm that stimulating 4 × 10 PBMCs in AIM-V medium supplemented with 10% FBS is the best approach and that HBeAg, HBsAg, and ALB are independent predictors of HBV-specific T-cell responses.
乙型肝炎病毒 (HBV) 的清除依赖于有效的适应性免疫反应，尤其是 HBV 特异性 T 细胞介导的细胞免疫；然而，很难有效地产生足够的 HBV 特异性 T 细胞。
在这项工作中，我们研究了刺激细胞、血清和培养基的比例作为三个主要因素，以确定最有效的程序，并将其应用于 HLA-A2(+)人群。同时，我们还检查了临床参数与 HBV 特异性免疫之间的相关性。关于扩增效率，4×10 个细胞刺激优于 2×10 个细胞刺激，AIM-V 培养基优于 1640 培养基，在可比条件下，胎牛血清 (FBS) 优于人 AB 血清。不出所料，该程序也适用于在 HLA-A2(+)个体中开发 HBV 特异性 CD8+T 细胞。扩增的 HBV 特异性 T 细胞反应随治疗时间而降低，与 HBV DNA 和 HBsAg 呈负相关。此外，HBV 特异性 IFN-γ+SFC 的数量与 ALT 水平呈强烈相关，与绝对淋巴细胞计数和 ALB 浓度呈负相关。
我们证实，在补充有 10%FBS 的 AIM-V 培养基中刺激 4×10 PBMC 是最佳方法，HBeAg、HBsAg 和 ALB 是 HBV 特异性 T 细胞反应的独立预测因子。
