优化的体外刺激可识别大多数慢性乙型肝炎患者的多功能HBV

StephenW

Optimized Ex Vivo Stimulation Identifies Multi-Functional HBV-specific T Cells in a Majority of Chronic Hepatitis B Patients
Conan G Chua  1   2 , Aman Mehrotra  1 , Tony Mazzulli  3   4 , David K Wong  1 , Jordan J Feld  1 , Harry L A Janssen  1 , Adam J Gehring  5   6
Affiliations

    PMID: 32647116 DOI: 10.1038/s41598-020-68226-5

Abstract

High antigen burden during chronic hepatitis B (CHB) results in a low frequency HBV-specific T cell response with restricted functionality. However, this observation is based on limited data because low T cell frequencies have hindered effective ex vivo analysis. We adapted the ELISpot assay to overcome this obstacle to measure ex vivo T cell responses in CHB patients. We modified the key variables of cell number and the peptide pulsing method to improve ex vivo detection of HBV-specific T cells. We detected IFN-γ responses in 10/15 vaccinated controls and 20/30 CHB patients, averaging 195 and 84 SFUs/2 × 106 PBMCs respectively. Multi-analyte FluoroSpots improved functional characterization of T cells. We detected IFN-γ responses in all tested vaccinated controls (n = 10) and CHB patients (n = 13). IL-2 responses were detectable in 9/10 controls and 10/13 patients. TNF-α displayed less sensitivity, detectable in only 7/10 controls and 7/13 patients. Antigen-specific analysis demonstrated that IFN-γ responses were dominated by polymerase and core, with weak responses to envelope and X. IL-2 responses were found in 3/5 patients and equally directed towards polymerase and core. While their ex vivo frequency is extremely low, a fraction of HBV-specific T cells are detectable and display multi-functionality ex vivo.

StephenW

优化的体外刺激可识别大多数慢性乙型肝炎患者的多功能HBV特异性T细胞
Conan G Chua 1 2，Aman Mehrotra 1，Tony Mazzulli 3 4，David K Wong 1，Jordan J Feld 1，Harry L A Janssen 1，Adam J Gehring 5 6
隶属关系

    PMID：32647116 DOI：10.1038 / s41598-020-68226-5

抽象

慢性乙型肝炎（CHB）期间的高抗原负担导致功能受限的低频HBV特异性T细胞反应。但是，此观察基于有限的数据，因为低T细胞频率阻碍了有效的离体分析。我们采用ELISpot分析法克服了这一障碍，以测量CHB患者的离体T细胞反应。我们修改了细胞数量和肽脉冲方法的关键变量，以改善离体检测HBV特异性T细胞的能力。我们在10/15的疫苗接种对照和20/30的CHB患者中检测到IFN-γ反应，分别平均195和84 SFU / 2×106 PBMC。多分析物FluoroSpots改善了T细胞的功能特性。我们在所有测试的接种对照（n = 10）和CHB患者（n = 13）中检测到IFN-γ反应。在9/10的对照组和10/13的患者中可检测到IL-2反应。 TNF-α敏感性较低，仅在7/10个对照和7/13个患者中可检测到。抗原特异性分析表明，IFN-γ反应以聚合酶和核心为主，对包膜和X的反应较弱。在3/5的患者中发现了IL-2反应，并且同样指向聚合酶和核心。尽管它们的离体频率极低，但一部分HBV特异性T细胞是可检测到的，并且离体显示出多功能。

StephenW

https://www.nature.com/articles/s41598-020-68226-5.pdf