使用广谱T细胞表位肽库和酶联免疫斑点分析对慢性乙型肝炎患者的HBV特异性T细胞反应性进行常规评估。
Routine evaluation of HBV-specific T cell reactivity in chronic hepatitis B using a broad-spectrum T-cell epitope peptide library and ELISpot assay.
作者信息
Wu Yandan, Liu Xiaotao, Mao Yuan, Ji Ruixue, Xia Lingzhi, Zhou Zining, Ding Yan, Li Pinqing, Zhao Yu, Peng Min, Qiu Jie, Shen Chuanlai
机构信息
Department of Microbiology and Immunology, Medical School of Southeast University, Nanjing, 210009, Jiangsu, China.
Nanjing KingMed Clinical Laboratory, Nanjing, 211899, Jiangsu, China.
出版信息
J Transl Med. 2024 Mar 11;22(1):266. doi: 10.1186/s12967-024-05062-5.
BACKGROUND
The clinical routine test of HBV-specific T cell reactivity is still limited due to the high polymorphisms of human leukocyte antigens (HLA) in patient cohort and the lack of universal detection kit, thus the clinical implication remains disputed.
METHODS
A broad-spectrum peptide library, which consists of 103 functionally validated CD8 T-cell epitopes spanning overall HBsAg, HBeAg, HBx and HBpol proteins and fits to the HLA polymorphisms of Chinese and Northeast Asian populations, was grouped into eight peptide pools and was used to establish an ELISpot assay for enumerating the reactive HBV-specific T cells in PBMCs. Totally 294 HBV-infected patients including 203 ones with chronic hepatitis B (CHB), 13 ones in acute resolved stage (R), 52 ones with liver cirrhosis (LC) and 26 ones with hepatocellular carcinoma (HCC) were detected, and 33 CHB patients were longitudinally monitored for 3 times with an interval of 3-5 months.
RESULTS
The numbers of reactive HBV-specific T cells were significantly correlated with ALT level, HBsAg level, and disease stage (R, CHB, LC and HCC), and R patients displayed the strongest HBV-specific T cell reactivity while CHB patients showed the weakest one. For 203 CHB patients, the numbers of reactive HBV-specific T cells presented a significantly declined trend when the serum viral DNA load, HBsAg, HBeAg or ALT level gradually increased, but only a very low negative correlation coefficient was defined (r = - 0.21, - 0.21, - 0.27, - 0.079, respectively). Different Nucleotide Analogs (NUCs) did not bring difference on HBV-specific T cell reactivity in the same duration of treatment. NUCs/pegIFN-α combination led to much more reactive HBV-specific T cells than NUCs monotherapy. The dynamic numbers of reactive HBV-specific T cells were obviously increasing in most CHB patients undergoing routine treatment, and the longitudinal trend possess a high predictive power for the hepatitis progression 6 or 12 months later.
CONCLUSION
The presented method could be developed into an efficient reference method for the clinical evaluation of cellular immunity. The CHB patients presenting low reactivity of HBV-specific T cells have a worse prognosis for hepatitis progression and should be treated using pegIFN-α to improve host T-cell immunity.
背景
由于患者群体中人类白细胞抗原（HLA）高度多态性以及缺乏通用检测试剂盒，HBV特异性T细胞反应性的临床常规检测仍然受限，因此其临床意义仍存在争议。
方法
一个广谱肽库由103个功能验证的CD8 T细胞表位组成，涵盖整个HBsAg、HBeAg、HBx和HBpol蛋白，适合中国及东北亚人群的HLA多态性，被分为八个肽池，并用于建立一种ELISpot检测法，以计数外周血单个核细胞（PBMC）中反应性HBV特异性T细胞。共检测了294例HBV感染患者，包括203例慢性乙型肝炎（CHB）患者、13例急性恢复期（R）患者、52例肝硬化（LC）患者和26例肝细胞癌（HCC）患者，并且对33例CHB患者进行了为期3次的纵向监测，间隔3至5个月。
结果
反应性HBV特异性T细胞数量与ALT水平、HBsAg水平及疾病阶段（R、CHB、LC和HCC）显著相关，R患者表现出最强的HBV特异性T细胞反应性，而CHB患者表现最弱。对于203例CHB患者，当血清病毒DNA载量、HBsAg、HBeAg或ALT水平逐渐升高时，反应性HBV特异性T细胞数量呈现显著下降趋势，但仅定义了非常低的负相关系数（分别为r = -0.21、-0.21、-0.27、-0.079）。在相同治疗持续时间内，不同的核苷类似物（NUC）对HBV特异性T细胞反应性没有差异。NUC/聚乙二醇干扰素-α联合治疗比NUC单药治疗产生更多反应性HBV特异性T细胞。大多数接受常规治疗的CHB患者中，反应性HBV特异性T细胞的动态数量明显增加，并且纵向趋势对6或12个月后的肝炎进展具有较高的预测能力。
结论
所提出的方法可发展成为一种用于细胞免疫临床评估的有效参考方法。HBV特异性T细胞反应性低的CHB患者肝炎进展预后较差，应使用聚乙二醇干扰素-α治疗以改善宿主T细胞免疫。
Zotero 插件