诺如病毒感染性腹泻患儿细胞免疫学特征分析
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摘要
[目的]诺如病毒(NV)是全球急性胃肠炎的主要原因,而感染诺如病毒的患者其黏膜和细胞免疫反应仍然知之甚少,本文旨在探讨诺如病毒感染性腹泻的患儿中,其细胞免疫学的特征。[方法]选取2017年6月～2018年1月在我院就诊或者住院的1个月～5岁的诺如病毒感染性腹泻患儿20例与健康的受试者11例,对于NV感染性腹泻的患者,在其出现临床症状之后的第2天、7天、14天、28天和第180天,分别采取患儿的唾液、粪便以及外周血单核细胞作为标本,收集健康受试者的唾液、粪便以及外周血单核细胞作为标本,按照相关的标准测定,并且将结果统计并分析。[结果]对于NV感染患儿,唾液和粪便中的NV特异性IgA水平在第14天达到峰值。粪便中NV特异性IgA的几何平均倍数增加(GMFR)高于唾液中NV特异性IgA。在第28天,在70%的感染者中观察到唾液中NV特异性IgA水平升高≥4倍。对于非特异性粪便IgA测定,所有感染受试者在第28天显示>4倍的变化。在受NV感染的受试者中,第7天的NV特异性粪便IgA水平与可检测到的病毒脱落持续时间之间存在负相关(Pearson r=-0.51;P=0.02)。在感染NV的受试者中,IgA和IgG ASC在感染NV之后第7天达到峰值,并在第14天显著降低。在受NV感染的受试者中,IgA ASC的水平在第7天与粪便中的峰值病毒载量相关(Pearson r=0.5;P=0.01)。在第14天观察到更大的相关性(Pearson r=0.72;P<0.0001),表明在第7天超过峰值水平的IgA ASC响应持续存在时,其病毒载量应该会更高。NV特异性记忆B细胞应答在第14天达到峰值并在第180天持续存在。从所有时间点的数据来看,NV特异性IgA记忆B细胞水平与唾液和粪便IgA滴度相关性良好(Pearson r分别为0.72和0.61)。[结论]唾液中NV特异性IgA和NV特异性记忆IgG细胞被鉴定为抗NV胃肠炎的新的相关物,所以了解黏膜、细胞和体液免疫的相对重要性对于诺如病毒疫苗的开发是非常重要的。
[Objective] Norovirus(NV) is the main cause of acute gastroenteritis in the world, and the mucosal and cellular immune responses in patients infected with Norovirus are still poorly understood. This article aimed to investigate the efficacy of Norovirus infectious diarrhea in children and the characteristics of cellular immunology.[Methods]Twenty patients with norovirus infectious diarrhea and 11 healthy volunteers(one month to five years old) were admitted to our hospital from June 2017 to January 2018. For patients with NV infectious diarrhea and healthy volunteers, On the 2 nd, 7 th, 14 th, 28 th, and 180 th days after infection, saliva, feces, and peripheral blood mononuclear cells were taken as samples and measured according to the relevant standards.[Results]For NV infected children, NV-specific IgA level in saliva and feces peaked on day 14. The geometric mean fold increase in NV-specific IgA in feces(GMFR) was higher than that in NV-specific IgA in saliva. On day 28, a 4 fold increase in NV-specific IgA level in saliva was observed in 70% of infected individuals. For the non-specific fecal IgA assay, all infected subjects showed a > 4-fold change on day 28. In NV-infected subjects, there was a negative correlation between day 7 NV-specific fecal IgA levels and detectable viral shedding duration(Pearson r=-0.51; P=0.02). In subjects infected with NV, IgA and IgG ASC peaked on the 7 th day after infection with NV and decreased significantly on the 14 th day. In NV-infected subjects, the level of IgA ASC was correlated with peak viral load in feces on day 7(Pearson r=0.5; P=0.01). A greater correlation was observed on day 14(Pearson r=0.72; P<0.0001), indicating that the viral load should be higher when IgA ASC responses persist beyond the peak level on day 7. The NV-specific memory B cell response peaked on day 14 and persisted on day 180 after infection with NV. From the data at all time points, there was a good correlation between NV-specific IgA memory B-cell levels and saliva and fecal IgA titers(Pearson r=0.72 and 0.61, respectively). [Conclusion]NV-specific IgA and NV-specific memory IgG cells in saliva were identified as novel related anti-NV gastroenteritis. In sum, understanding the relative importance of mucosal, cellular and humoral immunity is critically important for the development of norovirus vaccines.
[Objective] Norovirus(NV) is the main cause of acute gastroenteritis in the world, and the mucosal and cellular immune responses in patients infected with Norovirus are still poorly understood. This article aimed to investigate the efficacy of Norovirus infectious diarrhea in children and the characteristics of cellular immunology.[Methods]Twenty patients with norovirus infectious diarrhea and 11 healthy volunteers(one month to five years old) were admitted to our hospital from June 2017 to January 2018. For patients with NV infectious diarrhea and healthy volunteers, On the 2 nd, 7 th, 14 th, 28 th, and 180 th days after infection, saliva, feces, and peripheral blood mononuclear cells were taken as samples and measured according to the relevant standards.[Results]For NV infected children, NV-specific IgA level in saliva and feces peaked on day 14. The geometric mean fold increase in NV-specific IgA in feces(GMFR) was higher than that in NV-specific IgA in saliva. On day 28, a 4 fold increase in NV-specific IgA level in saliva was observed in 70% of infected individuals. For the non-specific fecal IgA assay, all infected subjects showed a > 4-fold change on day 28. In NV-infected subjects, there was a negative correlation between day 7 NV-specific fecal IgA levels and detectable viral shedding duration(Pearson r=-0.51; P=0.02). In subjects infected with NV, IgA and IgG ASC peaked on the 7 th day after infection with NV and decreased significantly on the 14 th day. In NV-infected subjects, the level of IgA ASC was correlated with peak viral load in feces on day 7(Pearson r=0.5; P=0.01). A greater correlation was observed on day 14(Pearson r=0.72; P<0.0001), indicating that the viral load should be higher when IgA ASC responses persist beyond the peak level on day 7. The NV-specific memory B cell response peaked on day 14 and persisted on day 180 after infection with NV. From the data at all time points, there was a good correlation between NV-specific IgA memory B-cell levels and saliva and fecal IgA titers(Pearson r=0.72 and 0.61, respectively). [Conclusion]NV-specific IgA and NV-specific memory IgG cells in saliva were identified as novel related anti-NV gastroenteritis. In sum, understanding the relative importance of mucosal, cellular and humoral immunity is critically important for the development of norovirus vaccines.
引文
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[9] Jain P,Jain A.Waterborne Viral Gastroenteritis:An Introduction to Common Agents[M]//Water and Health.Springer India,2014:53-74.
[10] Atmar RL,Opekun AR,Gilger MA,et al.Determination of the 50% human infectious dose for Norwalk virus\[J\].J Infect Dis,2014,209:1016-1022.
[11] Kelsall BL,Strober W.Chapter 24-Overview:Inductive and Effector Cells and Tissues of the Mucosal Immune System[J].Mucosal Immunol,2015:487-488.
[12] Aktar A,Rahman MA,Afrin S,et al.Plasma and memory B cell responses targeting O-specific polysaccharide(OSP)are associated with protection against Vibrio cholerae O1 infection among household contacts of cholera patients in Bangladesh[J].Plos Neglected Tropical Diseases,2018,12(4):e0006399.
[13] Devasia T,Lopman B,Leon J,et al.Association of host,agent and environment characteristics and the duration of incubation and symptomatic periods of norovirus gastroenteritis[J].Epidemiol Infect,2015,143(11):2308-2314.
[14] Mohn KG,Bredholt G,Brokstad KA,et al.Longevity of B-Cell and T-Cell Responses After Live Attenuated Influenza Vaccination in Children[J].J Infec Dis,2015,211(10):1541-1549.
[15] Treanor JJ,Atmar RL,Frey SE,et al.A novel intramuscular bivalent norovirus virus-like particle vaccine candidate-reactogenicity,safety,and immunogenicity in a phase 1 trial in healthy adults\[J\].J Infect Dis,2014,210(11):1763-1771.
[16] Kuebler PJ,Mehrotra ML,Mcconnell JJ,et al.Cellular immune correlates analysis of an HIV-1 preexposure prophylaxis trial[J].Proc Natl Acad Sci U S A,2015,112(27):8379-8384.
[2] Ramani S,Atmar RL,Estes MK.Epidemiology of human noroviruses and updates on vaccine development[J].Curr Opin Gastroenterol,2014,30(1):25-33.
[3] 潘秀珍.诺如病毒相关研究进展[J].医学研究生学报,2015,28(3):225-228.
[4] Debbink K,Lindesmith LC,Donaldson EF,et al.Emergence of new pandemic GII.4 Sydney norovirus strain correlates with escape from herd immunity[J].J Infect Dis,2013,208(11):1877-1887.
[5] Knight A,Li D,Uyttendaele M,et al.A critical review of methods for detecting human noroviruses and predicting their infectivity[J].Crit Rev Microbiol,2013,39(3):295-309.
[6] 王亚敬.诺罗病毒衣壳蛋白P结构域的结构和功能研究[D].南开大学,2014.
[7] Moore MD,Goulter RM,Jaykus LA.Human norovirus as a foodborne pathogen:challenges and developments[J].Annu Rev Food Sci Technol,2015,6(1):411-433.
[8] Martino BD,Profio FD,Felice ED,et al.Genetic heterogeneity of bovine noroviruses in Italy[J].Arch Virol,2014,159(10):2717-2722.
[9] Jain P,Jain A.Waterborne Viral Gastroenteritis:An Introduction to Common Agents[M]//Water and Health.Springer India,2014:53-74.
[10] Atmar RL,Opekun AR,Gilger MA,et al.Determination of the 50% human infectious dose for Norwalk virus\[J\].J Infect Dis,2014,209:1016-1022.
[11] Kelsall BL,Strober W.Chapter 24-Overview:Inductive and Effector Cells and Tissues of the Mucosal Immune System[J].Mucosal Immunol,2015:487-488.
[12] Aktar A,Rahman MA,Afrin S,et al.Plasma and memory B cell responses targeting O-specific polysaccharide(OSP)are associated with protection against Vibrio cholerae O1 infection among household contacts of cholera patients in Bangladesh[J].Plos Neglected Tropical Diseases,2018,12(4):e0006399.
[13] Devasia T,Lopman B,Leon J,et al.Association of host,agent and environment characteristics and the duration of incubation and symptomatic periods of norovirus gastroenteritis[J].Epidemiol Infect,2015,143(11):2308-2314.
[14] Mohn KG,Bredholt G,Brokstad KA,et al.Longevity of B-Cell and T-Cell Responses After Live Attenuated Influenza Vaccination in Children[J].J Infec Dis,2015,211(10):1541-1549.
[15] Treanor JJ,Atmar RL,Frey SE,et al.A novel intramuscular bivalent norovirus virus-like particle vaccine candidate-reactogenicity,safety,and immunogenicity in a phase 1 trial in healthy adults\[J\].J Infect Dis,2014,210(11):1763-1771.
[16] Kuebler PJ,Mehrotra ML,Mcconnell JJ,et al.Cellular immune correlates analysis of an HIV-1 preexposure prophylaxis trial[J].Proc Natl Acad Sci U S A,2015,112(27):8379-8384.