HBVsvp脉冲树突状细胞免疫疗法诱导Th1极化和乙型肝炎病毒特

StephenW

HBVsvp-Pulsed Dendritic Cell Immunotherapy Induces Th1 Polarization and Hepatitis B Virus-Specific Cytotoxic T Lymphocytes Production
Mohamed M S Farag  1 , Reda A Suef  1 , Ghada M Al-Toukhy  2 , Mohamed A Selim  1 , Mostafa A Elbahnasawy  1 , Nahla El Sharkawy  3 , Sameera Ezzat  4 , Nashwa Shebl  5 , Mohamed T M Mansour  6
Affiliations
Affiliations

    1
    Botany and Microbiology Department, Faculty of Science, Al-Azhar University, Cairo 11884, Egypt.
    2
    Virology & Immunology Department, Children's Cancer Hospital, Cairo 57357, Egypt.
    3
    Clinical Pathology Department, National Cancer Institute, Cairo University and Children Cancer Hospital, Cairo 57357, Egypt.
    4
    Epidemiology & Preventive Medicine Department, National Liver Institute, Menoufia University, Al Minufya, Egypt.
    5
    Hepatology Department, National Liver Institute, Menoufia University, Al Minufya, Egypt.
    6
    Virology & Immunology Department, National Cancer Institute, Cairo University and Children Cancer Hospital, Cairo 57357, Egypt.

    PMID: 32821133 PMCID: PMC7418458 DOI: 10.2147/IDR.S265681

Abstract

Background: In chronic hepatitis B virus (CHB) patients, both dendritic cells (DCs) and T cells are functionally impaired and consequently the HBV-specific cellular immune responses are downregulated. The present study aims to investigate whether monocyte-derived DC (MoDCs)-pulsed-HBV subviral particles (HBVsvp) can polarize Th1 cells to induce HBV-specific cytotoxic T-lymphocytes (CTL) responses in CHB patients.

Methods and materials: To this end, the human hepatoma HepG2.2.15 cell line was used to produce HBVsvp as a culturing system, and HBVsvp were concentrated for highly virus titer using the polyethylene glycol protocol. Peripheral blood mononuclear cells (PBMCs), collected from CHB patients and healthy donors, were differentiated into MoDCs and T cells. PBMCs-derived MoDCs were first pulsed with HBVsvp and then cultured with PBMCs-derived T cells. MoDCs and/or T subsets cells were identified for phenotypic activation by FACS analysis. The cytokine secretion of IL-4, IL-12, and IFN-γ in the culture supernatants was detected.

Results: The MoDCs were restored for their activation upon pulsing with HBVsvp in vitro, as identified by significantly overexpression of both CD86 and HLA-DR, and overproduction of IL-4 and IL-12. Furthermore, MoDCs-pulsed-HBVsvp induced Th1 frequencies and activated HBV-specific CTL to produce significantly highest amount of IFN-γ. Enhanced HBV-specific CTL led to strong cytolytic capacity against HepG2.2.15.

Conclusion: Overall, our data suggest that in vitro activation of MoDCs with HBVsvp overcomes the functionally impaired DCs and T cells in CHB patients offering a promising tool for therapeutic or vaccine-based approaches against HBV.

Keywords: CHB; HBV-specific CTL; MoDCs-pulsed-HBVsvp; immunotherapy.

© 2020 Farag et al.

StephenW

HBVsvp脉冲树突状细胞免疫疗法诱导Th1极化和乙型肝炎病毒特异性细胞毒性T淋巴细胞的产生
Mohamed M S Farag 1，Reda A Suef 1，Ghada M Al-Toukhy 2，Mohamed A Selim 1，Mostafa A Elbahnasawy 1，Nahla El Sharkawy 3，Sameera Ezzat 4，Nashwa Shebl 5，Mohamed T M Mansour 6
隶属关系
隶属关系

    1个
    Al-Azhar大学理学院植物学与微生物学系，埃及开罗11884。
    2
    儿童癌症医院病毒学和免疫学系，埃及开罗57357。
    3
    开罗大学国家癌症研究所临床病理科和儿童癌症医院，埃及开罗57357。
    4
    梅努菲亚大学国家肝脏研究所流行病学和预防医学系，埃及铝米努菲亚。
    5
    梅努非亚大学国家肝脏研究所肝病科，埃及铝米努菲亚。
    6
    开罗大学国家癌症研究所病毒与免疫学系和儿童癌症医院，埃及开罗57357。

    PMID：32821133 PMCID：PMC7418458 DOI：10.2147 / IDR.S265681

抽象

背景：在慢性乙型肝炎病毒（CHB）患者中，树突状细胞（DCs）和T细胞均功能受损，因此HBV特异性细胞免疫反应被下调。本研究旨在调查单核细胞衍生的DC（MoDCs）脉冲的HBV亚病毒颗粒（HBVsvp）是否能极化Th1细胞，从而在CHB患者中诱导HBV特异性细胞毒性T淋巴细胞（CTL）反应。

方法和材料：为此，使用人肝癌HepG2.2.15细胞系作为培养系统生产HBVsvp，并使用聚乙二醇规程将HBVsvp浓缩以获得高度病毒滴度。从CHB患者和健康供体中收集的外周血单个核细胞（PBMC）分化为MoDC和T细胞。首先将PBMC来源的MoDC用HBVsvp脉冲，然后用PBMC来源的T细胞培养。通过FACS分析鉴定出MoDC和/或T亚群细胞的表型活化。检测培养上清液中IL-4，IL-12和IFN-γ的细胞因子分泌。

结果：MoDCs在体外用HBVsvp脉冲后恢复了激活状态，这可以通过CD86和HLA-DR的显着过表达以及IL-4和IL-12的过量生产来确定。此外，MoDCs脉冲的HBVsvp诱导Th1频率并激活HBV特异性CTL，以产生明显最高量的IFN-γ。增强的HBV特异性CTL导致针对HepG2.2.15的强大溶细胞能力。

结论：总的来说，我们的数据表明，用HBVsvp在体外激活MoDCs可以克服CHB患者的功能受损的DC和T细胞，为治疗或基于疫苗的HBV方法提供了有希望的工具。

关键字：CHB; HBV特异性CTL； MoDCs-脉冲的HBVsvp；免疫疗法。

©2020 Farag等。

StephenW

https://www.dovepress.com/front_ ... ces-th1-polar_2.pdf