生物法合成傷寒O-糖蛋白結(jié)合疫苗及其免疫原性評估
發(fā)布時間：2019-06-24 13:36
【摘要】：傷寒由傷寒沙門氏菌(Salmonella Typhi)引發(fā),至今在發(fā)展中國家仍是備受關(guān)注的重要公共衛(wèi)生問題。文章通過敲除傷寒菌脂多糖合成途徑中O-抗原連接酶基因,轉(zhuǎn)入含腦膜炎奈瑟球菌(Neisseria meningitidis)蛋白糖基化途徑中糖基轉(zhuǎn)移酶的表達(dá)載體,以及改構(gòu)的重組銅綠假單胞菌(Pseudomonas Aeruginosa)外毒素A(r EPAN29)的表達(dá)載體,使細(xì)胞內(nèi)能夠誘導(dǎo)合成以傷寒O特異性多糖(O-specific polysaccharides,OPS)為目標(biāo)抗原、以r EPAN29為載體蛋白的傷寒OPS-r EPAN29糖蛋白復(fù)合物,并對純化所得復(fù)合物進(jìn)行了免疫原性評價。ELISA測定血清抗體滴度表明,r EPAN29作為載體蛋白能有效增加糖鏈的免疫原性,糖蛋白比單獨的多糖能誘導(dǎo)產(chǎn)生更好的免疫應(yīng)答;3次免疫、間隔3周比間隔2周Ig G滴度稍有提高;而免疫過量的糖蛋白,抗O-多糖的血清抗體效價并無提升。文章為生物法制備多糖-蛋白結(jié)合疫苗提供了新思路,理論上也適用于其他革蘭氏陰性菌的疫苗研發(fā)。
[Abstract]:Typhoid fever, caused by Salmonella typhimurium (Salmonella Typhi), is still an important public health problem in developing countries. In this paper, the O-antigen ligase gene in lipopolysaccharide synthesis pathway of typhoid typhimurium was knockout, the glycosyltransferase expression vector containing (Neisseria meningitidis) protein glycosylation pathway of Neisseria meningitis and the modified expression vector of recombinant Pseudomonas aeruginosa (Pseudomonas Aeruginosa) exotoxin A (r EPAN29 were transferred to induce the synthesis of typhoid O-specific polysaccharide (O-specific polysaccharides,OPS) as the target antigen in cells. The immunogenicity of typhoid OPS-r EPAN29 glycoprotein complex with r EPAN29 as carrier protein was evaluated. Elisa showed that r EPAN29 as carrier protein could effectively increase the immunogenicity of sugar chain, and glycoprotein could induce better immune response than polysaccharide alone. The titer of Ig G in 3 weeks was slightly higher than that in 2 weeks, but the titer of serum antibody against O-polysaccharide was not increased in the immunized glycoprotein and anti-O-polysaccharide. This paper provides a new idea for the preparation of polysaccharide-protein binding vaccine by biological method, and is also suitable for the vaccine development of other Gram-negative bacteria in theory.
【作者單位】： 吉首大學(xué)生物資源與環(huán)境科學(xué)學(xué)院;軍事醫(yī)學(xué)科學(xué)院生物工程研究所病原微生物生物安全國家重點實驗室;
【基金】：國家自然科學(xué)基金項目(編號:81373316)資助
【分類號】：R392
[Abstract]:Typhoid fever, caused by Salmonella typhimurium (Salmonella Typhi), is still an important public health problem in developing countries. In this paper, the O-antigen ligase gene in lipopolysaccharide synthesis pathway of typhoid typhimurium was knockout, the glycosyltransferase expression vector containing (Neisseria meningitidis) protein glycosylation pathway of Neisseria meningitis and the modified expression vector of recombinant Pseudomonas aeruginosa (Pseudomonas Aeruginosa) exotoxin A (r EPAN29 were transferred to induce the synthesis of typhoid O-specific polysaccharide (O-specific polysaccharides,OPS) as the target antigen in cells. The immunogenicity of typhoid OPS-r EPAN29 glycoprotein complex with r EPAN29 as carrier protein was evaluated. Elisa showed that r EPAN29 as carrier protein could effectively increase the immunogenicity of sugar chain, and glycoprotein could induce better immune response than polysaccharide alone. The titer of Ig G in 3 weeks was slightly higher than that in 2 weeks, but the titer of serum antibody against O-polysaccharide was not increased in the immunized glycoprotein and anti-O-polysaccharide. This paper provides a new idea for the preparation of polysaccharide-protein binding vaccine by biological method, and is also suitable for the vaccine development of other Gram-negative bacteria in theory.
【作者單位】： 吉首大學(xué)生物資源與環(huán)境科學(xué)學(xué)院;軍事醫(yī)學(xué)科學(xué)院生物工程研究所病原微生物生物安全國家重點實驗室;
【基金】：國家自然科學(xué)基金項目(編號:81373316)資助
【分類號】：R392
【參考文獻(xiàn)】
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