Table_1_A Co-culture Model of PBMC and Stem Cell Derived Human Nasal Epithelium Reveals Rapid Activation of NK and Innate T Cells Upon Influenza A Virus Infection of the Nasal Epithelium.pdf

Background: We established an in vitro co-culture model involving H3N2-infection of human nasal epithelium with peripheral blood mononuclear cells (PBMC) to investigate their cross-talk during early H3N2 infection.Methods: Nasal epithelium was differentiated from human nasal epithelial stem/progenitor cells and cultured wtih fresh human PBMC. PBMC and supernatants were harvested after 24 and 48 h of co-culture with H3N2-infected nasal epithelium. We used flow cytometry and Luminex to characterize PBMC subpopulations, their activation and secretion of cytokine and chemokines.Results: H3N2 infection of the nasal epithelium associated with significant increase in interferons (IFN-α, IFN-γ, IL-29), pro-inflammatory cytokines (TNF-α, BDNF, IL-3) and viral-associated chemokines (IP-10, MCP-3, I-TAC, MIG), detectable already after 24 h. This translates into rapid activation of monocytes, NK-cells and innate T-cells (MAIT and γδ T cells), evident with CD38+ and/or CD69+ upregulation.Conclusions: This system may contribute to in vitro mechanistic immunological studies bridging systemic models and possibly enable the development of targeted immunomodulatory therapies.

背景：本研究构建了一种体外共培养模型，涉及将H3N2病毒感染人鼻上皮细胞与外周血单个核细胞（PBMC）相结合，以研究其在早期H3N2感染过程中的相互作用。方法：通过分化人鼻上皮干细胞/祖细胞获得鼻上皮细胞，并与新鲜的人PBMC共同培养。在H3N2感染鼻上皮细胞后的24小时和48小时，收集PBMC和上清液。利用流式细胞术和Luminex技术对PBMC亚群及其激活状态以及细胞因子和趋化因子的分泌进行表征。结果：H3N2感染鼻上皮细胞后，与干扰素（IFN-α、IFN-γ、IL-29）、促炎细胞因子（TNF-α、BDNF、IL-3）和病毒相关趋化因子（IP-10、MCP-3、I-TAC、MIG）的显著增加相关，这些变化在感染后24小时即可检测到。这导致了单核细胞、自然杀伤细胞和先天T细胞（MAIT和γδ T细胞）的快速激活，表现为CD38+和/或CD69+表达上调。结论：该系统可能有助于体外机制性免疫学研究，架起全身性模型之间的桥梁，并可能促进靶向免疫调节疗法的开发。