Evaluation of stability and immune effects of three chitosan derivatives as adjuvants of nasal-spray hepatitis B vaccine
10.13200/j.cnki.cjb.004778
- VernacularTitle:3种壳聚糖衍生物作为鼻喷乙肝疫苗佐剂的稳定性及免疫效果评价
- Author:
Jilai LI
- Publication Type:Journal Article
- Keywords:
Nasal-spray vaccines;
Adjuvants;
Chitosan derivatives;
Immunogenicity
- From:
Chinese Journal of Biologicals
2026;39(10):1153-1159+1166
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare experimental nasal-spray hepatitis B vaccines with three chitosan(CS) derivatives, N-(2-hydroxyl) propyl-3-trimethyl ammonium chitosan chloride(HTCC), methylglycol chitosan(MGC) and chitosan acetate(CA), as adjuvants, and evaluate the stability and immunogenicity, so as to provide experimental basis for the development of mucosal hepatitis B vaccine.Methods HTCC, MGC, CA and CS were mixed with hepatitis B surface antigen(HBsAg) bulk solution respectively to prepare experimental nasal-spray hepatitis B vaccines(HTCC-HB, MGC-HB, CA-HB, and CS-HB groups) with final HBsAg concentration of 100 ??g/mL and adjuvant concentration of 1 mg/mL. The pH, particle size, Zeta potential and encapsulation efficiency of each group of vaccines were measured. The vaccines were stored at 2-8 ℃ for 4 and 40 weeks and at 37 ℃ for 4 weeks, with 0 week as the control. The stability was evaluated by detecting the variation of particle size. Female BAlB/c mice were immunized intranasally with four groups of experimental nasal-spray hepatitis B vaccines for 3 times at 0, 3,and 6 weeks, with 13 mice in each group. At the same time, HBsAg bulk solution group(S group), normal saline control group(NC group) and aluminum hydroxide adjuvant intramuscular injection group(Al-HB group) were set up. One week after the last immunization, enzyme-linked immunospot assay(ELISpot) was used to detect the numbers of spot forming cells(SFCs)secreting IFNγ, IL-2, and IL-4 in mouse spleen cells. Three weeks after the last immunization, ELISA was utilized to detect the levels of serum hepatitis B surface antibodies(anti-HBs) to calculate seroconversion rates and IgG2a/IgG1 ratios, and detect specific sIgA levels in bronchoalveolar lavage fluid(BAlF).Results The vaccine particle sizes of CS-HB and CA-HB groups were > 400 nm, and the encapsulation efficiencies were > 80%, whereas groups HTCC-HB and MGC-HB had particle sizes < 200 nm and encapsulation efficiencies < 20%. The particle sizes of HTCC-HB and MGC-HB changed only slightly after storage at 2-8 ℃ for 40 weeks and at 37 ℃ for 4 weeks. CS-HB decreased by 18. 2% after storage at 37 ℃ for 4 weeks.CA-HB increased by approximately 15. 6% after storage at 2-8 ℃ for 40 weeks and by approximately 131. 6% after storage at37 ℃ for 4 weeks. The seroconversion rates of mice in groups CS-HB, CA-HB, HTCC-HB, MGC-HB and Al-HB all reached100%, whereas no seroconversion occurred in groups S and NC. The SFC levels of IFNγ, IL-2, and IL-4 secreted by mouse splenocytes from groups CS-HB, CA-HB, HTCC-HB, MGC-HB and Al-HB were significantly higher than those in groups S and NC(P = 0. 000 2-0. 012 0). The mean IgG2a/IgG1 ratios in groups CS-HB, CA-HB, HTCC-HB and MGC-HB were all higher than that in group Al-HB, with only group MGC-HB showing a statistically significant difference(P = 0. 022 5). Specific sIgA was detectable in the BAlF of mice in the aforementioned groups, but not in groups Al-HB and S.Conclusion HTCC,MGC and CA, as adjuvants of experimental nasal-spray hepatitis B vaccine, can effectively induce humoral immunity, cellular immunity and mucosal immunity, and HTCC-HB and MGC-HB vaccines have smaller particle size and better stability,suggesting that the two CS derivatives, HTCC and MGC, have more potential as mucosal adjuvants of hepatitis B vaccines.
