Peptide-MHC cellular microarray with innovative data analysis system for simultaneously detecting multiple CD4 T-cell responses.
Peptide-MHC cellular microarray with innovative data analysis system for simultaneously detecting multiple CD4 T-cell responses.
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DOI：
10.1371/journal.pone.0011355
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发表时间：
2010-06-28
期刊：
影响因子：
3.7
通讯作者：
Kwok WW
中科院分区：
文献类型：
--
作者：
Ge X;Gebe JA;Bollyky PL;James EA;Yang J;Stern LJ;Kwok WW
肽：MHC 细胞微阵列已被提议用于同时表征多个 Ag 特异性 T 细胞群。使用该工具研究对复杂病原体的免疫反应的实践需要对 T 细胞表位的广泛了解和用于阵列制造的肽：MHC 复合物的可用性以及用于结果解释的专门数据分析方法。我们将肽：DR0401 复合物、抗 CD28、抗 CD11a 和细胞因子捕获抗体共同固定在室载玻片的表面上，以生成一个功能阵列，能够从先前引发的体外 T 细胞培养物中检测稀有的 Ag 特异性 T 细胞群。还开发了一种新颖的统计方法，以促进将原始阵列类数据批量处理为标准化终点分数，该分数与总 Ag 特异性 T 细胞输入线性相关。应用这些方法来分析甲型流感病毒抗原特异性 T 细胞反应，我们不仅揭示了最突出的病毒表位，而且还证明了健康个体中抗病毒细胞反应的异质性。应用这些方法来检查产生胰岛素的β细胞自身抗原特异性T细胞反应，我们观察到自身免疫性糖尿病患者和健康个体之间几乎没有差异，这表明糖尿病状态和外周自身反应性T细胞之间存在更微妙的关联。该数据分析系统对于T细胞特异性和功能测试是可靠的。肽：MHC 细胞微阵列可用于在各种转化环境中使用有限的血液样本获得多参数结果。
Peptide:MHC cellular microarrays have been proposed to simultaneously characterize multiple Ag-specific populations of T cells. The practice of studying immune responses to complicated pathogens with this tool demands extensive knowledge of T cell epitopes and the availability of peptide:MHC complexes for array fabrication as well as a specialized data analysis approach for result interpretation. We co-immobilized peptide:DR0401 complexes, anti-CD28, anti-CD11a and cytokine capture antibodies on the surface of chamber slides to generate a functional array that was able to detect rare Ag-specific T cell populations from previously primed in vitro T cell cultures. A novel statistical methodology was also developed to facilitate batch processing of raw array-like data into standardized endpoint scores, which linearly correlated with total Ag-specific T cell inputs. Applying these methods to analyze Influenza A viral antigen-specific T cell responses, we not only revealed the most prominent viral epitopes, but also demonstrated the heterogeneity of anti-viral cellular responses in healthy individuals. Applying these methods to examine the insulin producing beta-cell autoantigen specific T cell responses, we observed little difference between autoimmune diabetic patients and healthy individuals, suggesting a more subtle association between diabetes status and peripheral autoreactive T cells. The data analysis system is reliable for T cell specificity and functional testing. Peptide:MHC cellular microarrays can be used to obtain multi-parametric results using limited blood samples in a variety of translational settings.
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作者：
Newell, Evan W.;Klein, Lawrence O.;Yu, Wong;Davis, Mark M.
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