目的 制备黏膜免疫佐剂Chi-CpG NP，探究其对结核分枝杆菌ESAT-6疫苗免疫保护效应的增强作用。方法 采用离子交联法制备Chi-CpG NP并检测其理化性质；将雌性BALB/c小鼠随机分为PBS对照组、ESAT-6组、ESAT-6+CpG组、ESAT-6+Chi NP组、ESAT-6+Chi-CpG NP组，经鼻腔黏膜免疫后，于第42天检测小鼠体液免疫（IgG、IgG1、gG2a、sIgA）和细胞免疫水平[γ干扰素（IFN-γ）、白细胞介素（IL）-4、IL-17A]；经腹腔注射结核分枝杆菌H37Rv进行攻毒实验，感染后4周观察肺组织病理损伤程度。结果 成功制备的Chi-CpG NP呈规则球形，有较好的蛋白吸附功能。免疫后42d，ESAT-6+Chi-CpG NP组小鼠血清IgG、IgG1、IgG2a及鼻、阴道黏膜sIgA水平均显著高于其它实验组（P＜0.05），脾细胞上清中IFN-γ、IL-2、IL-17A分泌水平均为各组最高（P＜0.05）；此外，ESAT-6+Chi-CpG NP组肺组织炎性浸润程度最轻。结论 Chi-CpG NP可有效增强ESAT-6疫苗诱导的体液免疫、黏膜免疫及细胞免疫应答，显著提升小鼠对结核分枝杆菌感染的抵抗能力。

结核分枝杆菌；ESAT-6抗原；黏膜免疫佐剂；Chi-CpG NP

[1]Kisimba K,Kasumbwe K,Odun-Ayo F,et al.Flavonoids:Potential Novel Inhibitors of Mycobacterium tuberculosis[J].Infect Disord Drug Targets,2025,25(8):e18715265361578.

[2]Ferluga J,Yasmin H,Bhakta S,et al.Vaccination Strategies Against Mycobacterium tuberculosis:BCG and Beyond[J].Adv Exp Med Biol,2021,1313:217-240.

[3]Bouzeyen R,Javid B.Therapeutic Vaccines for Tuberculosis:An Overview[J].Front Immunol,2022,13:878471.

[4]Singh DK,Ahmed M,Akter S,et al.Prevention of tuberculosis in cynomolgus macaques by an attenuated Mycobacterium tuberculosis vaccine candidate[J].Nat Commun,2025,16(1):1957.

[5]李玉洁,余海燕,杨雨婷,等.结核分枝杆菌分泌蛋白早期分泌性抗原6(ESAT-6)的免疫学性质及其在新型疫苗中作用的研究进展[J].细胞与分子免疫学杂志,2024,40(01):89-94.

[6]Blanco FC,Vázquez CL,García EA,et al.Mycobacterium bovis mutant in the virulence factors PhoP,ESAT-6 and CFP-10 persisted in mouse organs after a year post-vaccination[J].Tuberculosis(Edinb),2024,149:102574.

[7]Wang Y,Liu S,Li B,et al.A novel CpG ODN compound adjuvant enhances immune response to spike subunit vaccines of porcine epidemic diarrhea virus[J].Front Immunol,2024,15:1336239.

[8]Li Z,Li Y,Wang Y,et al.Intranasal immunization with a rNMB0315 and combination adjuvants induces protective immunity against Neisseria meningitidis serogroup B in mice[J].Int Immunopharmacol,2021,93:107411.

[9]王丽梅.结核病黏膜疫苗的研究进展[J].中国人兽共患病学报,2022,38(02):170-174,181.

[10]李文桂,陈雅棠.载体介导的结核分枝杆菌ESAT-6疫苗的研制现状[J].热带医学杂志,2024,24(02):299-304.

[11]Jiang T,Ji H,Zhang L,et al.Chitosan Oligosaccharide Exerts Anti-Allergic Effect against Shrimp Tropomyosin-Induced Food Allergy by Affecting Th1 and Th2 Cytokines[J].Int Arch Allergy Immunol,2019,180(01):10-16.

[12]White AD,Tran AC,Sibley L,et al.Spore-FP1 tuberculosis mucosal vaccine candidate is highly protective in guinea pigs but fails to improve on BCG-conferred protection in non-human primates[J].Front Immunol,2023,14:1246826.

[13]Slovakova M,Janovska S,Sleha R,et al.Nasal Immunization Using Chitosan Nanoparticles with Glycoprotein B of Murine Cytomegalovirus[J].J Microbiol Biotechnol,2024,34(3):663-672.

[14]Chuang CC,Tsai MH,Yen HJ,et al.A fucoidan-quaternary chitosan nanoparticle adjuvant for anthrax vaccine as an alternative to CpG oligodeoxynucleotides[J].Carbohydr Polym,2020,229:115403.

[15]Li Z,He Y,Deng L,et al.A fast-dissolving microneedle array loaded with chitosan nanoparticles to evoke systemic immune responses in mice[J].J Mater Chem B,2020,8(2):216-225.

[16]邢佳音,赵祥祥,孙铭瑞,等.疫苗佐剂研究进展[J].中国生物工程杂志,2025,45(10):64-76.