Hagen Jodi, Houchins Jeffrey P, Kalyuzhny Alexander E
R&D Systems, Inc, Minneapolis, MN, USA.
Methods Mol Biol. 2011;682:207-15. doi: 10.1007/978-1-60327-409-8_15.
Enzyme-Linked Immuno Spot (ELISpot) assay is widely used for vaccine development, cancer and AIDS research, and autoimmune disease studies. The output of an ELISpot assay is a formation of colored spots which appear at the sites of cells releasing cytokines, with each individual spot representing a single cytokine-releasing cell.We have shown that hydrogen peroxide-induced oxidative stress was causing ∼twofold decrease in the number of lymphocytes secreting the TH1 cytokines IFN-gamma and IL-2, as well as chemokine IL-8 and cytokine TNF alpha. However, the number of cells secreting TH2 cytokines IL-4 and IL-5 in hydrogen peroxide-treated group did not change. Our ELISpot data indicate that oxidative stress may affect TH1-TH2 cytokine secretion balance which, in turn, may underlie developments of various pathological conditions. We adopted ELISpot assay for studying oxidative stress in human peripheral blood lymphocytes by analyzing the acute effect of hydrogen peroxide treatment on the frequency of cells secreting IFN-gamma, IL-2, IL-4, IL-5, IL-8, and TNF-alpha.
酶联免疫斑点（ELISpot）测定法广泛应用于疫苗研发、癌症与艾滋病研究以及自身免疫性疾病研究。ELISpot测定法的结果是在释放细胞因子的细胞部位形成有色斑点，每个单独的斑点代表一个分泌细胞因子的细胞。我们已经表明，过氧化氢诱导的氧化应激导致分泌TH1细胞因子IFN-γ和IL-2以及趋化因子IL-8和细胞因子TNF-α的淋巴细胞数量减少约两倍。然而，过氧化氢处理组中分泌TH2细胞因子IL-4和IL-5的细胞数量没有变化。我们的ELISpot数据表明，氧化应激可能影响TH1-TH2细胞因子分泌平衡，这反过来可能是各种病理状况发展的基础。我们采用ELISpot测定法，通过分析过氧化氢处理对分泌IFN-γ、IL-2、IL-4、IL-5、IL-8和TNF-α的细胞频率的急性影响，来研究人外周血淋巴细胞中的氧化应激。
