In situ Activated Intestinal T Cells Expanded in vitro – without Addition of Antigen – Produce IFN-γ and IL-10 and Preserve Their Function during Growth
{"title":"In situ Activated Intestinal T Cells Expanded in vitro – without Addition of Antigen – Produce IFN-γ and IL-10 and Preserve Their Function during Growth","authors":"J. Agnholt, K. Kaltoft","doi":"10.1159/000049200","DOIUrl":null,"url":null,"abstract":"Objective: The balance between mucosal CD4+ T cells producing IFN-γ or IL-10 is essential in the maintenance of intestinal homeostasis. We aimed to investigate how in situ activated T cells were expanded in vitro according to a new cell culture protocol and if it selected for continuous clonal CD4+ T cells capable of producing mainly IFN-γ or IL-10. Methods: T cell cultures were established from colonic biopsy specimens of 27 patients with Crohn’s disease and from 10 healthy controls in a medium supplemented with IL-2 and IL-4 but without addition of exogenous antigen or mitogen. Cytokine production was measured after stimulation (IL-12, super antigen) and inhibition (ciclosporin and methylprednisolone). Results: Cytokine production was increased in cultures from patients with Crohn’s disease compared to controls (IFN-γ, p = 0.005; IL-10, p = 0.003; TNF-α, p = 0.03). Early cultures were highly responsive to IL-12 stimulation. We established CD4+ T-cell clones escaping cellular senescence with an inflammatory or regulatory cytokine profile. Discussion: The data indicate that cultures of in situ activated inflammatory and regulatory subpopulations of intestinal T lymphocytes with pathogenic importance in Crohn’s disease can be established preserving their functional properties during growth.","PeriodicalId":77124,"journal":{"name":"Experimental and clinical immunogenetics","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2002-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000049200","citationCount":"18","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Experimental and clinical immunogenetics","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1159/000049200","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 18
Abstract
Objective: The balance between mucosal CD4+ T cells producing IFN-γ or IL-10 is essential in the maintenance of intestinal homeostasis. We aimed to investigate how in situ activated T cells were expanded in vitro according to a new cell culture protocol and if it selected for continuous clonal CD4+ T cells capable of producing mainly IFN-γ or IL-10. Methods: T cell cultures were established from colonic biopsy specimens of 27 patients with Crohn’s disease and from 10 healthy controls in a medium supplemented with IL-2 and IL-4 but without addition of exogenous antigen or mitogen. Cytokine production was measured after stimulation (IL-12, super antigen) and inhibition (ciclosporin and methylprednisolone). Results: Cytokine production was increased in cultures from patients with Crohn’s disease compared to controls (IFN-γ, p = 0.005; IL-10, p = 0.003; TNF-α, p = 0.03). Early cultures were highly responsive to IL-12 stimulation. We established CD4+ T-cell clones escaping cellular senescence with an inflammatory or regulatory cytokine profile. Discussion: The data indicate that cultures of in situ activated inflammatory and regulatory subpopulations of intestinal T lymphocytes with pathogenic importance in Crohn’s disease can be established preserving their functional properties during growth.
目的:粘膜CD4+ T细胞产生IFN-γ或IL-10之间的平衡在维持肠道稳态中至关重要。我们的目的是研究原位活化T细胞是如何根据新的细胞培养方案在体外扩增的,以及它是否被选择为能够主要产生IFN-γ或IL-10的连续克隆CD4+ T细胞。方法:选取27例克罗恩病患者和10例健康对照者的结肠活检标本,在添加IL-2和IL-4但不添加外源抗原或丝裂原的培养基中培养T细胞。在刺激(IL-12,超抗原)和抑制(环孢素和甲基强的松龙)后测量细胞因子的产生。结果:与对照组相比,克罗恩病患者培养物中细胞因子的产生增加(IFN-γ, p = 0.005;IL-10, p = 0.003;TNF-α, p = 0.03)。早期培养物对IL-12刺激反应强烈。我们建立了CD4+ t细胞克隆逃避细胞衰老与炎症或调节细胞因子谱。讨论:这些数据表明,在克罗恩病中具有致病性重要性的肠道T淋巴细胞原位激活炎症和调节亚群的培养可以在生长过程中保持其功能特性。