金黃色葡萄球菌IsdB、TRAP和ClfA融合蛋白的免疫保護(hù)作用
發(fā)布時間：2019-02-11 07:36
【摘要】：金黃色葡萄球菌(Staphylococcus aureus,S.aureus)是人和動物的一種重要致病菌。隨著S.aureus耐藥性菌株的不斷出現(xiàn),使S.aureus感染的治療越來越困難,因此疫苗接種便成為防治S.aureus感染的重要手段。近年來人們已經(jīng)研究了S.aureus的許多結(jié)構(gòu)蛋白和致病因子作為疫苗候選抗原,如IsdB、IsdA、Trap、Rap、ClfA、FnbP和Cna等。雖然這些候選抗原都能誘導(dǎo)機(jī)體產(chǎn)生較好的免疫應(yīng)答,但免疫保護(hù)效果仍不夠理想。為了進(jìn)一步提高候選抗原的免疫保護(hù)作用,本研究將免疫原性較好的S.aureus蛋白IsdB、Trap和ClfA進(jìn)行融合表達(dá),并對表達(dá)產(chǎn)物的免疫應(yīng)答和免疫保護(hù)作用進(jìn)行研究,為S.aureus疫苗研制提供新的候選抗原。 本研究首先對ClfA N2N3及其分段后的ClfA_(N2)和ClfA_(N3)進(jìn)行表達(dá),經(jīng)純化后分別與弗氏佐劑進(jìn)行乳化,并免疫小鼠研究其免疫保護(hù)作用。結(jié)果,三種蛋白均可以引起小鼠產(chǎn)生良好的免疫應(yīng)答,用S. aureus Newman株和HLJ23-1株分別對免疫小鼠攻毒后,ClfA_(N3)、ClfA_(N2)蛋白對小鼠的免疫保護(hù)作用與ClfA_(N2N3)蛋白免疫相近。選取ClfA_(N3)與IsdB_(id)和TRAP進(jìn)行融合表達(dá),表達(dá)的融合蛋白命名為ITC,并將此融合蛋白與弗氏佐劑乳化作為免疫原免疫接種小鼠,研究其免疫保護(hù)作用。結(jié)果,ITC免疫可以引起小鼠產(chǎn)生良好的的免疫應(yīng)答;分別用S. aureus的Newman株、HLJ23-1株和Wood46株進(jìn)行攻毒后,重組ITC蛋白免疫組小鼠存活數(shù)高于其他各組。重組ITC蛋白具有更好的免疫保護(hù)作用,可以作為預(yù)防S. aurues感染的候選疫苗抗原。
[Abstract]:Staphylococcus aureus (Staphylococcus aureus,S.aureus) is an important pathogen in humans and animals. With the emergence of drug-resistant strains of S.aureus, the treatment of S.aureus infection becomes more and more difficult, so vaccination has become an important means to prevent and cure S.aureus infection. In recent years, many structural proteins and pathogenic factors of S.aureus have been studied as vaccine candidate antigens, such as IsdB,IsdA,Trap,Rap,ClfA,FnbP and Cna. Although these candidate antigens can induce a better immune response, the immune protection effect is still not satisfactory. In order to further improve the immune protection of candidate antigens, IsdB,Trap and ClfA, which have good immunogenicity, were fused and expressed, and the immune response and protective effect of the expressed products were studied. To provide a new candidate antigen for the development of S.aureus vaccine. In this study, ClfA N2N3 and its segmented ClfA_ (N2) and ClfA_ (N3) were first expressed, then purified and emulsified with Freund's adjuvant respectively. The results showed that all of the three proteins could induce a good immune response in mice. ClfA_ (N3) was induced by S. aureus Newman strain and HLJ23-1 strain, respectively. The immune protective effect of ClfA_ (N2) protein was similar to that of ClfA_ (N2N3) protein. ClfA_ (N3) was fused with IsdB_ (id) and TRAP. The fusion protein was named ITC, and emulsified with Freund's adjuvant to immunize mice. The results showed that the mice immunized with ITC could produce a good immune response, and the survival rate of the mice immunized with recombinant ITC protein was higher than that of the other groups after inoculation with Newman, HLJ23-1 and Wood46 strains of S. aureus. The recombinant ITC protein has better immune protection and can be used as a candidate vaccine antigen for the prevention of S. aurues infection.
【學(xué)位授予單位】：黑龍江八一農(nóng)墾大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R378;S852.611
本文編號：2419525
[Abstract]:Staphylococcus aureus (Staphylococcus aureus,S.aureus) is an important pathogen in humans and animals. With the emergence of drug-resistant strains of S.aureus, the treatment of S.aureus infection becomes more and more difficult, so vaccination has become an important means to prevent and cure S.aureus infection. In recent years, many structural proteins and pathogenic factors of S.aureus have been studied as vaccine candidate antigens, such as IsdB,IsdA,Trap,Rap,ClfA,FnbP and Cna. Although these candidate antigens can induce a better immune response, the immune protection effect is still not satisfactory. In order to further improve the immune protection of candidate antigens, IsdB,Trap and ClfA, which have good immunogenicity, were fused and expressed, and the immune response and protective effect of the expressed products were studied. To provide a new candidate antigen for the development of S.aureus vaccine. In this study, ClfA N2N3 and its segmented ClfA_ (N2) and ClfA_ (N3) were first expressed, then purified and emulsified with Freund's adjuvant respectively. The results showed that all of the three proteins could induce a good immune response in mice. ClfA_ (N3) was induced by S. aureus Newman strain and HLJ23-1 strain, respectively. The immune protective effect of ClfA_ (N2) protein was similar to that of ClfA_ (N2N3) protein. ClfA_ (N3) was fused with IsdB_ (id) and TRAP. The fusion protein was named ITC, and emulsified with Freund's adjuvant to immunize mice. The results showed that the mice immunized with ITC could produce a good immune response, and the survival rate of the mice immunized with recombinant ITC protein was higher than that of the other groups after inoculation with Newman, HLJ23-1 and Wood46 strains of S. aureus. The recombinant ITC protein has better immune protection and can be used as a candidate vaccine antigen for the prevention of S. aurues infection.
【學(xué)位授予單位】：黑龍江八一農(nóng)墾大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R378;S852.611
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1 馮昊;劉樂鋒;遲佳琦;王寧;李閏婷;佟春玉;馬金柱;朱戰(zhàn)波;崔玉東;;金黃色葡萄球菌凝集因子A的免疫原性[J];生物工程學(xué)報;2009年08期
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1 王寧;金黃色葡萄球菌IsdB3與TRAP融合蛋白免疫保護(hù)作用研究[D];黑龍江八一農(nóng)墾大學(xué);2009年
，本文編號：2419525
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