Optimization of culture conditions for HBV-specific T cell expansion in vitro from chronically infected patients.

IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY BMC Biotechnology Pub Date : 2024-10-14 DOI:10.1186/s12896-024-00908-8
Li Wang, Hongjiao Chen, Yuanqi Yang, Ying Huang, Weixian Chen, Di Mu
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Abstract

Background: 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.

Results: 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 × 105 cells stimulation was superior to 2 × 106 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.

Conclusions: We confirm that stimulating 4 × 105 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.

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优化慢性感染患者 HBV 特异性 T 细胞体外扩增的培养条件。
背景:乙型肝炎病毒(HBV)的清除依赖于有效的适应性免疫反应,特别是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、血清和培养基这三个主要因素的比例,以确定最有效的程序,并将其应用于 HLA-A2 (+) 人群。同时,我们还研究了临床参数与 HBV 特异性免疫之间的相关性。在扩增效率方面,4 × 105 细胞刺激优于 2 × 106 细胞刺激,AIM-V 培养基优于 1640 培养基,胎牛血清(FBS)优于同等条件下的人 AB 血清。正如预期的那样,这种方法也适用于培养 HLA-A2(+) 人的 HBV 特异性 CD8 + T 细胞。扩大的 HBV 特异性 T 细胞反应随治疗时间的延长而减少,并与 HBV DNA 和 HBsAg 呈负相关。此外,HBV 特异性 IFN-γ + SFCs 的数量与 ALT 水平密切相关,与淋巴细胞绝对计数和 ALB 浓度呈负相关:我们证实,在添加了 10% FBS 的 AIM-V 培养基中刺激 4 × 105 PBMCs 是最佳方法,HBeAg、HBsAg 和 ALB 是预测 HBV 特异性 T 细胞反应的独立指标。
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来源期刊
BMC Biotechnology
BMC Biotechnology 工程技术-生物工程与应用微生物
CiteScore
6.60
自引率
0.00%
发文量
34
审稿时长
2 months
期刊介绍: BMC Biotechnology is an open access, peer-reviewed journal that considers articles on the manipulation of biological macromolecules or organisms for use in experimental procedures, cellular and tissue engineering or in the pharmaceutical, agricultural biotechnology and allied industries.
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