Kirkwood Carl D, Boniface Karen, Richardson Simone, Taraporewala Zenobia F, Patton John T, Bishop Ruth F
Enteric Virus Research Group, Murdoch Childrens Research Institute, Royal Children's Hospital, Melbourne, Australia.
J Med Virol. 2008 Jun;80(6):1090-8. doi: 10.1002/jmv.21160.
Rotaviruses are the single most important causes of severe acute diarrhoea in children worldwide. Despite success in developing vaccines, there is still a lack of knowledge about many components of the immune response, particularly those to non-structural proteins. This study established radioimmunoprecipitation (RIP) assays using labeled G1P[8], G2P[4], and G4P[6] human rotaviruses to examine the spectrum and duration of rotavirus antibodies in sera collected sequentially for 18-36 months from 27 children after hospitalization for primary rotavirus gastroenteritis. Five children experienced rotavirus re-infections. Primary responses detected to non-structural protein NSP2 declined to baseline after 100-150 days. Responses were heterotypic between NSP2 of G1P[8] and G4P[8] rotaviruses. Re-infections after 465-786 days boosted antibody levels to NSP2of both serotypes, together with the appearance of anti-NSP2 to G2P[4], even though there was no evidence of infection with this serotype. We developed an enzyme-immunoassay to measure sequential levels of anti-NSP2 IgG and IgA, using recombinant (heterotypic) NSP2 derived from SA11 (G3P[2]). Anti-NSP2 IgG and IgA were detected in sera from 23/23 (100%) and 18/24 (75%) of children after primary infection, declined to baseline after 100-150 days, were boosted after rotavirus re-infections, and again declined to baseline 150 days later. Anti-NSP2 IgA was also detected after primary infection, in duodenal juice from 14/16 (87%), and faecal extract from 11/19 (57%) of children. Sequential estimation of anti-NSP2 EIA levels in sera could be a sensitive index of rotavirus infection and re-infection. The potential of anti-NSP2 to limit viral replication after re-infection deserves further study.
轮状病毒是全球范围内导致儿童严重急性腹泻的最重要单一病因。尽管在开发疫苗方面取得了成功,但对于免疫反应的许多组成部分,尤其是针对非结构蛋白的免疫反应,仍缺乏了解。本研究建立了放射性免疫沉淀(RIP)测定法,使用标记的G1P[8]、G2P[4]和G4P[6]人轮状病毒,来检测27名因原发性轮状病毒肠胃炎住院后连续18至36个月采集的血清中轮状病毒抗体的谱和持续时间。5名儿童经历了轮状病毒再次感染。对非结构蛋白NSP2检测到的初次反应在100 - 150天后降至基线。G1P[8]和G4P[8]轮状病毒的NSP2之间的反应是异型的。465 - 786天后的再次感染使两种血清型的NSP2抗体水平升高,同时出现了针对G2P[4]的抗NSP2抗体,尽管没有证据表明感染了该血清型。我们开发了一种酶免疫测定法,使用源自SA11(G3P[2])的重组(异型)NSP2来测量抗NSP2 IgG和IgA的连续水平。初次感染后,23/23(100%)的儿童血清中检测到抗NSP2 IgG,18/24(75%)的儿童血清中检测到抗NSP2 IgA,它们在100 - 150天后降至基线,轮状病毒再次感染后升高,150天后再次降至基线。初次感染后,14/16(87%)的儿童十二指肠液和11/19(57%)的儿童粪便提取物中也检测到抗NSP2 IgA。血清中抗NSP2酶免疫测定水平的连续估计可能是轮状病毒感染和再次感染的敏感指标。抗NSP2在再次感染后限制病毒复制的潜力值得进一步研究。