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- Title
副猪嗜血杆菌病四价灭活苗的制备及对豚鼠的免疫效果评价.
- Authors
程家园; 王 迪; 李 亮; 占松鹤; 邢 刚; 刘雪兰; 孙 裴; 魏建忠; 李 郁
- Abstract
To provide a technical reserve for the development of a new tetravalent inactivated vaccine against Haemophilus parasuis (HPS) disease (Glässer's disease), four strains of HPS (serotypes 4, 5, 7 and 13, codenamed AT-2, SX-1P, HF-1P and HQ-2) were screened and used as vaccine strains, and three kinds of tetravalent inactivated vaccine (named V-1, V-2 and V-3) were prepared with the concentration of 1.0×109 cfu/mL, 2.0×109 cfu/mL and 4.0×109 cfu/mL following by inactivated with formaldehyde and mixed with mineral oil adjuvant ISA 201VG. The guinea pigs were used as experimental animal models and divided into four immune groups (I- IV group) injected with V-1, V-2, V-3 and commercial vaccine (serotypes 4, 5), respectively, at dose of 4.0× 109 cfu/mL to evaluate the efficacy. The indirect ELISA was used for detecting the level of IgG antibody in serum, serum bactericidal test (SBT) for detecting the phagocytic activity of functional antibody, MTT assay for detecting the proliferation of spleen lymphocyte, and ELISA for detecting the levels of cytokines (IL-4, IL-10, IFN-γ, TNF- β and MCP-1). The immune protection rate was measured by intraperitoneal challenge, the bacteria load in tissues of guinea pigs in each group was measured by plate counting method to evaluate the colonization number of challenge strains, and pathological sections of lung, liver, spleen and kidney were prepared and observed. The results showed that V-1, V-2 and V-3 fully qualified for physical properties, sterility and safety testing. During the whole immune cycle, there were no significant changes in the detection indexes of the blank control group. The serum IgG antibody titers in immune groups I-IV were 1:12800, the level of IgG increased continuously and there was no significant difference among the four groups (P>0.05), the corresponding serum collected had strong bactericidal effect. The levels of IL-4, IL-10, IFN-γ, TNF-β and MCP-1 in serum increased continuously and had no significant difference among the four groups (P>0.05). The overall protective rate of I-IV was 85%, 100%, 100% and 60%, respectively, after intraperitoneal challenge; The amount of bacteria colonized in all organs (lung, liver, kidney and spleen) of immune group I-III was significantly lower than that of immune group IV (P<0.05), and the pathological changes of each organ were milder than those in immune group IV. In conclusion, the V-1, V-2 and V-3 could evoke good humoral and cellular immunity in guinea pigs, and among which V-2 and V-3 could resist 100% of lethal doses of HPS serotypes 4, 5, 7 and 13. This study verified for the first time that V-2 and V-3 can be used as candidates of tetravalent inactivated (serotypes 4, 5, 7 and 13) vaccine against Haemophilus parasuis disease, which lay the foundation for the vaccine development of Glässerʼs disease.
- Subjects
HAEMOPHILUS diseases; PATHOLOGICAL physiology; GUINEA pigs; CELLULAR immunity; MINERAL oils; LUNGS; IMMUNOGLOBULINS
- Publication
Chinese Journal of Preventive Veterinary Medicine / Zhongguo Yufang Shouyi Xuebao, 2022, Vol 44, Issue 10, p1091
- ISSN
1008-0589
- Publication type
Article
- DOI
10.3969/j.issn.1008-0589.202112013