Mechanism of immunopotentiator CVC1302 regulating somatic hypermutation

DU Luping; LU Haiyan; HOU Liting; YU Xiaoming; CHENG Haiwei; ZHANG Yuanpeng; CHEN Jin; ZHENG Qisheng; HOU Jibo

doi:10.13300/j.cnki.hnlkxb.2025.01.023

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Mechanism of immunopotentiator CVC1302 regulating somatic hypermutation

Animal science·Veterinary medicine | 更新时间：2026-11-05

- Mechanism of immunopotentiator CVC1302 regulating somatic hypermutation
- Journal of Huazhong Agricultural University Vol. 44, Issue 1, Pages: 211-216(2025)
- 作者机构：
  
  1.江苏省农业科学院动物免疫工程研究所/国家兽用生物制品技术创新中心/ 江苏省食品质量安全重点实验室，南京 210014
  2.兽用生物制品（泰州）国泰技术创新中心，泰州 225300
- 作者简介：
- 基金信息：
- DOI：10.13300/j.cnki.hnlkxb.2025.01.023
  CLC： S852.4
- Received：15 December 2023，
  
  Published：15 January 2025
- 稿件说明：
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杜露平，鲁海燕，侯立婷，等.免疫增强剂CVC1302调控体细胞高频突变的机制研究［J］.华中农业大学学报，2025，44（1）：211‐216.

DU Luping,LU Haiyan,HOU Liting,et al.Mechanism of immunopotentiator CVC1302 regulating somatic hypermutation[J].Journal of Huazhong Agricultural University,2025,44(01):211-216.
杜露平，鲁海燕，侯立婷，等.免疫增强剂CVC1302调控体细胞高频突变的机制研究［J］.华中农业大学学报，2025，44（1）：211‐216. DOI： 10.13300/j.cnki.hnlkxb.2025.01.023.

DU Luping,LU Haiyan,HOU Liting,et al.Mechanism of immunopotentiator CVC1302 regulating somatic hypermutation[J].Journal of Huazhong Agricultural University,2025,44(01):211-216. DOI： 10.13300/j.cnki.hnlkxb.2025.01.023.

摘要

为探究CVC1302调控生发中心B细胞发生体细胞高频突变的免疫机制，本研究将4-羟基-3-硝基苯乙酰基耦联鸡卵白蛋白（NP-OVA）混合免疫增强剂CVC1302后，利用ISA206乳化获得疫苗。BALB/c小鼠分为2组，分别后腿肌肉注射NP-ISA206和NP-CVC1302-ISA206，每只50 μg NP-OVA，免疫后14 d，利用流式分选获得生发中心B细胞，利用巢氏PCR扩增B细胞免疫球蛋白序列可变区VH

186.2

，Western blot检测诱导活化的胞苷脱氨酶（AID）、Pax5表达水平，β-actin作为内参，比较组间AID、Pax5表达差异；利用荧光定量PCR检测

AID、Pax5

基因转录水平。试验结果显示：CVC1302显著诱导生发中心B细胞免疫球蛋白序列VH

186.2

突变频率，试验组W33L突变频率为62.2%，而对照组仅为20.25%；CVC1302可提升AID蛋白、Pax5蛋白表达水平，其中试验组AID蛋白相对表达水平为0.72，对照组仅为0.16，试验组Pax5蛋白相对表达水平为0.62，对照组仅为0.26；CVC1302增强

AID

基因、

Pax5

基因转录水平，相较于对照组分别提高2.36、4.13倍。推断CVC1302依赖Pax5介导AID表达，调控生发中心B细胞发生体细胞高频突变。

Abstract

ISA206 was used to obtain the vaccine by emulsifying 4-hydroxy-3-nitrophenylacetyl conjugated chicken egg white protein （NP-OVA） with immunopotentiator CVC1302 to study the immune mechanism of CVC1302 regulating the occurrence of somatic hypermutation in germinal center B （

GC B） cells. Six-week-old BALB/c female mice were divided into 2 groups and immunized with NP-CVC1302-ISA206， NP-ISA206 in the hind leg muscles， with each mouse receiving 100 μL vaccine and 50 μg NP-OVA. Germinal center B cells were obtained with flow cytometry after14 days of immunization， and the variable region VH

186.2

gene sequence of immunoglobulin in GC B cells was amplified with nested-PCR. Western blot was used to detect the expression level of cytidine deaminase （AID） and Pax5 induced activation， and the differences in the expression level of AID and Pax5 between groups were compared with β-actin as an internal reference. The transcription level of

AID

and

Pax5

in GC B cells was analyzed with fluorescence quantitative PCR. The results showed that immunopotentiator CVC1302 significantly induced the mutation frequency of VH

186.2

in the immunoglobulin sequence of germinal center B cells， with a W33L mutation frequency of 62.2% in the NP-CVC1302 immunized group and only 20.25% in the NP immunized group. CVC1302 increased the expression level of AID protein and Pax5 protein in GC B cells. The relative expression level of AID protein in the NP-CVC1302 immunized group was 0.72， while that in the NP immunized group was only 0.16. The relative expression level of Pax5 protein in the NP-CVC1302 immunized group was 0.62， while that in the NP immunized group was only 0.26. CVC1302 enhanced the transcription level of

AID

gene and

Pax5

gene in the NP-CVC1302 immunized group， which was increased by 2.36 and 4.13 times compared to that in the NP immunized group. It is indicated that CVC1302 relies on Pax5 mediated AID expression to regulate the occurrence of somatic hypermutation in germinal center B cells.

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references

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