携带白细胞介素-2/肝细胞生长因子4-kringle拮抗剂双基因的减毒沙门菌对小鼠免疫功能的影响  

The effects of attenuated Salmonella carrying IL⁃2/NK4 double gene on immune function in mice

作　　者：杨志华 蒋如如 牛廷献 郭晓宇 宋薇 张萍 张媛媛 哈小琴 YANG Zhihua;JIANG Ruru;NIU Tingxian;GUO Xiaoyu;SONG Wei;ZHANG Ping;ZHANG Yuanyuan;HA Xiaoqin(The 940 Hospital of the Joint Logistics Support Force of Chinese People's Liberation Army,Lanzhou 730050,Gansu Province,China)

机构地区：[1]中国人民解放军联勤保障部队第九四〇医院,甘肃兰州730050

出　　处：《中国生物制品学杂志》2024年第5期571-576,共6页Chinese Journal of Biologicals

基　　金：甘肃省重点研发计划-社会发展类项目(17YF1FA135);中央高校基本科研项目(31920200020);兰州市人才创新项目(2016-RC-61).

摘　　要：目的研究携带白细胞介素-2(interleukin-2,IL⁃2)/肝细胞生长因子4-kringle拮抗剂(4-kringle antagonist of hepatocyte growth factor,NK4)双基因的减毒沙门菌(Ty21a-pIRES-IL-2-NK4,命名为TPIN)对小鼠体液及细胞免疫功能的影响。方法将18只BALB/c小鼠(雌雄各半)随机分为对照(胃管饲服1.5 mL 10%NaHCO3)、Ty21a(胃管饲服0.1 mL Ty21a)、TPIN(胃管饲服0.1 mL TPIN)组,6只/组。初次免疫7 d后加强免疫,共2次,给药第21天,经眼球采血分离血清,ELISA法检测血清IgG抗体水平;无菌分离小鼠胸腺和脾脏,体外分别用Ty21a、TPIN刺激培养的小鼠脾细胞,72 h后,CCK-8法检测小鼠脾细胞增殖能力,ELISA法检测小鼠脾细胞细胞因子表达水平;称重小鼠脾脏和胸腺,计算脾和胸腺指数,并进行脾和胸腺组织HE染色。结果与对照和Ty21a组相比,TPIN组小鼠血清IgG水平(F=111.74,P<0.01)以及脾细胞上清IFNγ、IL-4和IL-10含量均明显升高(F分别为38.21、11.37、26.92,P均<0.05),脾指数和胸腺指数明显升高(F分别为10.419、5.859,P均<0.05)。Ty21a、TPIN组小鼠胸腺组织皮质增宽,淋巴细胞增多,有轻度炎性反应;脾脏组织白髓增多,淋巴细胞增多伴随中性粒细胞浸润。结论TPIN具有良好的免疫保护作用,能明显刺激机体产生体液免疫和细胞免疫,从而对肿瘤可能具有较好的治疗作用。Objective To study the effects of attenuated Salmonella(Ty21a-pIRES-IL-2-NK4,TPIN)carrying interleukin-2(IL⁃2)/4-kringle antagonist of hepatocyte growth factor(NK4)double gene on humoral and cellular immune function.Methods Eighteen BALB/c mice,half male and half female,were randomly divided into control group(1.5 mL 10%NaHCO3 by gastric tube feeding),Ty21a group(0.1 mL Ty21a by gastric tube feeding)and TPIN group(0.1 mL TPIN by gastric tube feeding),with 6 mice in each group.The immunization was boosted twice 7 d after the initial immunization.At 21d after administration,the blood samples were collected from eyeballs and the serum was separated,which was detected for the serum IgG antibody level by ELISA.The thymus and spleen of mice were isolated aseptically,and the spleen cells were stimulated by Ty21a and TPIN respectively in vitro.After 72 h,the proliferation ability of spleen cells was measured by CCK-8 assay,and the expression level of cytokines in spleen cells was detected by ELISA.The spleen and thymus were weighed,the spleen and thymus indexes were analyzed,and HE staining was performed.Results Compared with the control group and Ty21a group,the serum IgG level(F=111.74,P<0.01)and the contents of IFNγ,IL-4 and IL-10 in spleen cell supernatant(F=38.21,11.37 and 26.92,respectively,each P<0.05)increased significantly,as well as the spleen and thymus indexes(F=10.419 and 5.859,respectively,each P<0.05)showed significant increase.In mice of Ty21a and TPIN group,the thymus cortex widened,lymphocytes increased,and there was mild inflammatory reaction;the white pulp and lymphocytes in spleen increased with neutrophil infiltration.Conclusion TPIN has a good immune protective effect,and can significantly stimulate the body to produce humoral immunity and cellular immunity,which may have a good therapeutic effect on tumors.

关 键 词：白细胞介素-2 肝细胞生长因子抑制剂4 减毒沙门菌 体液免疫 细胞免疫 脾指数 胸腺指数 

分 类 号：R392.1[医药卫生—免疫学]