新疆荒漠肉苁蓉粗多糖对口蹄疫疫苗抗体和T细胞亚群的影响

doi:10.13560/j.cnki.biotech.bull.1985.2020-1455

摘要/Abstract

摘要：

为了筛选口蹄疫（foot-and-mouth disease,FMD）疫苗的新型多糖佐剂,选用新疆荒漠肉苁蓉粗多糖与口蹄疫灭活抗原（inactivated foot-and-mouth disease viral antigen,FMDV-Ag）制备成水佐剂疫苗,ISA-206油佐剂疫苗和无佐剂疫苗,通过肌肉途径免疫ICR小鼠,检测免疫后血清抗体水平、淋巴细胞增殖反应、CD3⁺CD4⁺、CD3⁺CD8⁺和CD4⁺CD44⁺、CD8⁺CD44⁺T细胞亚群的表达水平等评价疫苗的免疫增强效果,并且监测小鼠免疫后的行为和体重变化观察安全性。肉苁蓉粗多糖可以显著提高FMDV特异性的抗体水平（P<0.05）;同时显著增强了FMDV特异性的淋巴细胞增殖水平（P<0.05）,显著促进了CD4⁺、CD8⁺和CD44⁺T细胞比例（P<0.05）。免疫后小鼠没有观察到异常行为,且对小鼠体重增长没有影响。肉苁蓉粗多糖作为FMDV-Ag的水佐剂通过增强体液和细胞免疫水平发挥良好的免疫促进作用。

关键词: 新疆荒漠肉苁蓉, 粗多糖, 口蹄疫疫苗, 佐剂

Abstract:

To screen new polysaccharide vaccine adjuvants of foot-and-mouth disease,the crude polysaccharide from Cistanche deserticola in Xinjiang（CDCP）and inactivated foot-and-mouth disease viral antigen（FMDV-Ag）were selected to prepare water adjuvant vaccines,ISA-206 oil adjuvant vaccine and adjuvant-free vaccine. ICR mice were immunized by muscle. Serum antibody level,lymphocyte proliferation response,and expressions of CD3⁺CD4⁺,CD3⁺CD8⁺ and CD4⁺CD44⁺,CD8⁺CD44⁺T cell subsets were measured to evaluate the immune enhancement effect after the vaccination,and the behaviors and weights of the mice were monitored for observing the safety of the vaccines. The results showed CDCP substantially enhanced the FMDV-specific antibody level（P<0.05）and significantly increased lymphocyte proliferative responses（P<0.05）. T responses were also significantly enhanced（P<0.05）,including proliferative responses of CD4⁺,CD8⁺,and CD44⁺ T cells. No abnormal behavior for mice after immunization was observed and the weights of mice also were not affected. CDCP as water adjuvant of FMDV-Ag has a fine immune-promoting effect by enhancing humoral and cellular immune response.

Key words: Cistanche deserticola in Xinjiang, crude polysaccharide, foot-and-mouth disease viral vaccine, adjuvant

张爱莲, 巴雪丽, 王丹阳, 赵兵. 新疆荒漠肉苁蓉粗多糖对口蹄疫疫苗抗体和T细胞亚群的影响[J]. 生物技术通报, 2021, 37(9): 212-218.

ZHANG Ai-lian, BA Xue-li, WANG Dan-yang, ZHAO Bing. Effects of Crude Polysaccharide from Cistanche deserticola in Xinjiang on Foot-and-Mouth Disease Viral Vaccine Antibody and T cell Subgroup[J]. Biotechnology Bulletin, 2021, 37(9): 212-218.

图/表 5

图1 间接ELISA检测血清中FMDV特异性抗体水平 CDCP 配伍FMDV-Ag 肌肉免疫小鼠后,采集初次免疫后14 d、21 d 和28 d小鼠血清,ELISA 法检测FMDV 特异性的抗体水平（n=6）。与FMDV-Ag 组相比较,* 表示P < 0.05,** 表示P < 0.01,***表示P < 0.001。下同

Fig.1 Detection on FMDV-specific antibody by indirect ELISA Mice were intramuscularly immunized with CDCP and FMDV-Ag,on 14,28 and 42 d after primary immunization,sera were collected. The level of FMDV-specific antibody was detected by ELISA（n=6）. *P < 0.05,**P < 0.01 and ***P < 0.001 compared with FMDV-Ag group. The same below

图2 CDCP对淋巴细胞增殖的影响 CDCP配伍FMDV-Ag肌肉免疫小鼠后,初免后21 d制备小鼠脾细胞悬液,MTT法检测CDCP对脾淋巴细胞的增殖作用（n=6）

Fig. 2 Effect of CDCP on splenocyte proliferation Mice were intramuscularly immunized with CDCP and FMDV-Ag,splenocyte suspension was isolated on 21 d after primary immunization,and the effects of CPCD on splenocyte proliferation were determined by MTT assay（n=6）

图3 CDCP对CD4+和CD8+ T细胞亚群的影响 CDCP配伍FMDV-Ag免疫小鼠21 d后,流式细胞术检测CDCP对小鼠脾脏中CD4+和CD8+ T细胞的影响（n=6）

Fig.3 Effects of CDCP on CD4+ and CD8+ T cell subsets Mice were immunized with CDCP and FMDV-Ag on 21 d after primary immunization,cells from spleen of mice were used to detect T cell subsets by flow cytometry（n=6）

图4 CDCP对CD4+ CD44+、CD8+ CD44+效应T细胞的影响流式细胞术检测初免后21 d,CDCP对小鼠脾脏中CD4+ CD44+、CD8+ CD44+ 效应T细胞的影响（n=6）

Fig.4 Effects of CDCP on CD4+ CD44+ and CD8+ CD44+ effector T cell On 21 d after primary immunization,flow cytometry was used to detect the effects of CDCP on CD4+CD44+ and CD8+CD44+ effector T cell in spleen（n=6）

表1 FMDV/CDCP免疫后小鼠体重变化

Table 1 Changes in body weight of mice immunized with FMDV/CDCP

组别 Group	每组小鼠数 Number of each group（n）	体重 Weight/g
组别 Group	每组小鼠数 Number of each group（n）	0 d	14 d	21 d	28 d
0.9%NaCl	5	21.60±1.01	27.20±2.52	31.72±2.85	32.51±3.01
FMDV-Ag	5	23.22±1.21	28.68±2.34	32.54±1.87	34.18±2.72
FMDV-Ag/CDCP	5	22.68±0.37	27.26±2.28	31.72±2.55	34.44±2.73
FMDV/ISA-206	5	21.38±1.04	27.58±1.44	30.56±1.28	32.96±3.01

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