Quantitative analysis of anti-hepatitis C virus anti body-secreting B cells in patients with chronic hepatitis C
Quantitative analysis of anti-hepatitis C virus anti body-secreting B cells in patients with chronic hepatitis C
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DOI：
10.1002/hep.20917
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发表时间：
2006-01-01
期刊：
影响因子：
13.5
通讯作者：
Alter, HJ
中科院分区：
文献类型：
--
作者：
Umemura, T;Wang, RYH;Alter, HJ
为探讨丙型肝炎患者体液免疫的定量特征，建立了酶联免疫斑点法（ELISpot）检测抗丙型肝炎病毒（HCV）分泌型B细胞的方法。受试者工作特征曲线分析表明，检测B细胞对NS 5 b、NS 3、E2和核心抗原的应答的特异性为100%，灵敏度为58%至92%。在39例慢性丙型肝炎患者中，抗-HCV分泌性B细胞对所有检测的HCV抗原的中位总和显著较高（47.3个斑点形成细胞[SFC]/10（6）个外周血单核细胞[PBMC]）（15.3 SFC/10（6）PBMC; P = 0.05）或11个未感染对照（5-3 SFC/10（6）PBMC; P <0.001）;慢性与恢复患者的显著差异（P = 0.018）是对非结构抗原NS 3和NS 5 b的反应性。抗-HCV免疫球蛋白M（IgM）分泌B细胞也很容易检测到，并持续数十年的HCV感染，有IgM阳性细胞之间的慢性和恢复患者没有差异。基因型1、2和3的患者对基因型1衍生抗原的ELISpot反应性是等同的。抗-HCV IgG分泌B细胞数与血清转氨酶及循环抗体水平呈显著相关。总之，ELISpot测定法可适用于研究B细胞以及T细胞对HCV的应答。单细胞水平的测量表明，体液免疫在HCV感染的恢复中起次要作用，而B细胞免疫在持续感染者中最强。
To investigate the quantitative characteristics of humoral immunity in patients with hepatitis C, we established an enzyme-linked immunosorbent spot (ELISpot) assay for detection of anti-hepatitis C virus (HCV)-secreting B cells. Receiver operating characteristic curve analysis demonstrated 100% specificity and 58% to 92% sensitivity for detecting B-cell responses to NS5b, NS3, E2, and core antigens. The median sum of anti-HCV-secreting B cells to all HCV antigens tested was significantly higher in 39 patients with chronic hepatitis C (47.3 spot forming cells [SFCs]/10(6) peripheral blood mononuclear cells [PBMCs]) than in 9 recovered subjects (15.3 SFCs/10(6) PBMCs; P = .05) or 11 uninfected controls (5-3 SFCs/10(6) PBMCs; P < .001); the significant difference (P = .018) in chronic versus recovered patients was in reactivity to nonstructural antigens NS3 and NS5b. Anti-HCV immunoglubulin M (IgM)-secreting B cells were also readily detected and persisted decades into HCV infection; there was no difference in IgM-positive cells between chronic and recovered patients. ELISpot reactivity to genotype 1-derived antigens was equivalent in patients of genotypes 1, 2, and 3. There was significant correlation between the numbers of anti-HCV IgG-secreting B cells and serum aminotransferase and to the level of circulating antibody. In conclusion, ELISpot assays can be adapted to study B-cell as well as T-cell responses to HCV. Measurement at the single-cell level suggests that humoral immunity plays a minor role in recovery from HCV infection and that B-cell immunity is strongest in those with persistent infection.