小鼠诺如病毒感染实验小鼠的抗体变化规律分析

doi:10.12300/j.issn.1674-5817.2020.127

摘要/Abstract

摘要： 目的研究KM、BALB/c、NIH、C57BL/6J及BALB/c-nu共5个品系小鼠感染小鼠诺如病毒（murine norovirus,MNV）后总IgG抗体及中和抗体的变化规律。方法收集KM、BALB/c、NIH、C57BL/6J及BALB/c-nu共5个品系小鼠,每个品系分为对照组和感染组,感染组灌胃接种MNV液,对照组灌胃同等体积的生理盐水（即0.9%NaCl溶液）。分别在感染后第1~9周采血分离血清,ELISA法测定总IgG抗体水平（即吸光度值）,RAW264.7细胞病变法测定中和抗体滴度（即半数保护剂量,PD₅₀）。t检验比较感染组和对照组在每个时间点的总IgG抗体吸光度均值差异;单因素方差分析比较各品系小鼠感染组总抗体及中和抗体滴度,分析各品系小鼠的抗体变化规律。结果经t检验,KM、BALB/c、NIH、C57BL/6J及BALB/c-nu小鼠分别在第21、35、14、14及28天感染组总IgG抗体吸光度均值显著高于对照组（P<0.01）。各品系小鼠的总IgG抗体水平相比,NIH>C57BL/6J>KM>BALB/c>BALB/c-nu,且BALB/c及BALB/c-nu总抗体吸光度均值显著低于NIH和C57BL/6J小鼠（P<0.05）;NIH、C57BL/6J、KM及BALB/c小鼠感染第6周时抗体水平升高较快,NIH小鼠第49天进入平台期,其余3个品系小鼠在实验周期内总IgG抗体水平仍处于上升趋势,其中BALB/c-nu小鼠总抗体水平最低。各品系小鼠的中和抗体滴度比较,C57BL/6J>NIH>KM>BALB/c>BALB/c-nu,其lgPD₅₀均值分别为3.881、3.819、3.746、3.146及2.338;经方差分析显示,BALB/c及BALB/c-nu小鼠中和抗体滴度显著低于C57BL/6J、NIH及KM小鼠（P<0.01）。除了BALB/c-nu小鼠外,其余4个品系小鼠感染第2周时中和抗体滴度升高较快,BALB/c-nu小鼠中和抗体一直处于较低水平。结论 NIH、C57BL/6J及KM小鼠感染MNV的体液免疫应答较强,BALB/c及BALB/c-nu小鼠体液免疫应答较弱。

关键词: 小鼠诺如病毒, 感染, 抗体变化规律, 小鼠

Abstract: Objective To study the changes of total IgG antibodies and neutralizing antibodies in KM, BALB/c, NIH, C57BL/6J and BALB/c-nu mice infected with murine norovirus (MNV). Methods Five strains of KM, BALB/c, NIH, C57BL/6J and BALB/c-nu mice were collected. Each strain was divided into control group and infection group. The infection group was inoculated with MNV by gavage, and the control group was inoculated with the same volume of normal saline. Blood and serum were collected from 1 to 9 weeks after infection, total IgG antibody level (mean absorbance) was determined by ELISA, and neutralizing antibody titer (half protective dose, PD₅₀) was determined by RAW264.7 cytopathic method. T-test was used to compare the mean absorbance difference between the infection group and the control group at each time point; One way ANOVA was used to compare the total antibody and neutralizing antibody titer of each strain of the infection group, and to analyze the antibody changes of each strain of mice. Results T-test showed that the mean absorbance of total IgG antibody in KM, BALB/c, NIH, C57BL/6J and BALB/c-nu mice infected on the day 21, 35, 14, 14 and 28 was significantly higher than that of the control group (P < 0.01). To compared the total IgG antibody level of each strain of mice, the results showed NIH>C57BL/6J>KM>BALB/c>BALB/c-nu. The total IgG antibody levels of BALB/c and BALB/c-nu mice were significantly lower than that of NIH and C57BL/6J mice (P < 0.05); The antibody level of NIH, C57BL/6J, KM and BALB/c mice increased rapidly at the 6th week of infection, and NIH mice entered the plateau stage at the 49th day. The IgG antibody of the other three strains was still on the rise during the experimental period, and the overall antibody level of BALB-/c-nu mice was the lowest. The overall neutralizing antibody titers of each strain of mice were compared, showing that C57BL/6J>NIH>KM>BALB/c>BALB/c-nu, and the mean values of lgPD₅₀ were 3.881, 3.819, 3.746, 3.146, and 2.338, respectively. Analysis of variance showed that neutralizing antibody titer of BALB/c and BALB/c-nu mice was significantly lower than that of C57BL/6J, NIH and KM mice (P < 0.01). Except for BALB/c-nu mice, the neutralizing antibody of the other four strains increased rapidly at the second week of infection, and the neutralizing antibody of BALB/c-nu mice was always at a low level. Conclusion The humoral immune response of NIH, C57BL/6J and KM mice infected with MNV is stronger, and that of BALB/c and BALB/c-nu mice is weaker.

Key words: Murine norovirus, Infection, Antibody changes, Mice

中图分类号:

Q95-33
R-332

李晓波, 付瑞, 王淑菁, 李威, 王莎莎, 黄宗文, 秦骁, 王吉, 岳秉飞. 小鼠诺如病毒感染实验小鼠的抗体变化规律分析[J]. 实验动物与比较医学, 2021, 41(4): 343-350.

LI Xiaobo, FU Rui, WANG Shujing, LI Wei, WANG Shasha, HUANG Zongwen, QIN Xiao, WANG Ji, YUE Bingfei. Analysis of Antibody Changes in Laboratory Mice Infected with Murine Norovirus[J]. Laboratory Animal and Comparative Medicine, 2021, 41(4): 343-350.

图/表 3

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