CAJ | 학술논문

目的评价二价轮状病毒灭活疫苗免疫效果,探讨其开发的可行性.方法首先将G1型和G3型轮状病毒灭活疫苗等量混合制备二价疫苗,设立抗原量相等的G1型、G3型单价疫苗对照及PBS阴性对照,然后肌内注射免疫小鼠,通过ELISA检测血清和肠道中病毒特异性IgG和IgA、微量中和实验检测血清中病毒特异性中和抗体,ELISPOT分析免疫小鼠脾细胞中病毒特异性IFN-γ分泌细胞和IL-4分泌细胞的数量,评价二价疫苗的免疫效果.结果与阴性对照组相比,二价疫苗显著诱发了G1和G3特异性血清IgG、IgA、中和抗体和肠道IgG、IgA.与单价疫苗组相比,二价疫苗诱发的G1特异性血清IgG和IgA、肠道IgG和IgA及血清中和抗体与G1单价疫苗组诱发的相应抗体间差异无统计学意义；但二价疫苗组诱发的G3特异性血清IgG(t=2.691,P＜0.05)及中和抗体(t=2.561,P＜0.05)却显著高于G3单价疫苗组诱发的相应抗体水平,其余G3特异性抗体水平两者无差异.同时,与阴性对照组相比,二价疫苗免疫小鼠脾细胞中IFN-γ和IL-4分泌细胞的数量均显著增加.与单价疫苗组相比,G1刺激产生的IFN-γ和IL-4分泌细胞的数量,二价疫苗组与G1单价疫苗组无显著差异；但在二价疫苗组,G3刺激产生的IL-4分泌细胞的数量显著高于G3单价疫苗组(t=2.327,P＜0.05),而IFN-γ分泌细胞的数量两组无显著性差异.结论二价疫苗可以有效的诱发免疫应答,疫苗不同成分诱发的免疫应答未发生干扰抑制,这些结果为临床二价轮状病毒灭活疫苗的开发提供了实验依据.
목적 평개이개륜상병독멸활역묘면역효과,탐토기개발적가행성.방법 수선장G1형화G3형륜상병독멸활역묘등량혼합제비이개역묘,설립항원량상등적G1형、G3형단개역묘대조급PBS음성대조,연후기내주사면역소서,통과ELISA검측혈청화장도중병독특이성IgG화IgA、미량중화실험검측혈청중병독특이성중화항체,ELISPOT분석면역소서비세포중병독특이성IFN-γ분비세포화IL-4분비세포적수량,평개이개역묘적면역효과.결과 여음성대조조상비,이개역묘현저유발료G1화G3특이성혈청IgG、IgA、중화항체화장도IgG、IgA.여단개역묘조상비,이개역묘유발적G1특이성혈청IgG화IgA、장도IgG화IgA급혈청중화항체여G1단개역묘조유발적상응항체간차이무통계학의의；단이개역묘조유발적G3특이성혈청IgG(t=2.691,P＜0.05)급중화항체(t=2.561,P＜0.05)각현저고우G3단개역묘조유발적상응항체수평,기여G3특이성항체수평량자무차이.동시,여음성대조조상비,이개역묘면역소서비세포중IFN-γ화IL-4분비세포적수량균현저증가.여단개역묘조상비,G1자격산생적IFN-γ화IL-4분비세포적수량,이개역묘조여G1단개역묘조무현저차이；단재이개역묘조,G3자격산생적IL-4분비세포적수량현저고우G3단개역묘조(t=2.327,P＜0.05),이IFN-γ분비세포적수량량조무현저성차이.결론 이개역묘가이유효적유발면역응답,역묘불동성분유발적면역응답미발생간우억제,저사결과위림상이개륜상병독멸활역묘적개발제공료실험의거.
Objective To evaluate the immune effects of bivalent inactivated rotavirus vaccine (IRV) and investigate the viability of development of bivalent IRV.Methods Firstly,bivalent IRV was prepared by mixing G1 IRV and G3 IRV with equal amount,G1 IRV and G3 IRV as monovalent control,PBS as negative control.Secondly,those vaccines were vaccinated to the mice by intramuscular injection.Then,to evaluate the immune effects of bivalent IRV,the levels of serum or fecal rotavirus specific IgG and IgA were assessed by ELISA,the levels of serum neutralized antibody were measured by microneutralization assay,the number of IFN-γ or IL-4 secreting cells were analyzed by ELISPOT assay.Results Compared to negative control group,bivalent IRV induced the higher levels of serum and fecal G1 and G3 rotavirus specific antibody.It was found that there were no significant differences for the levels of serum IgG and IgA,fecal IgG and IgA,serum neutralized antibody between induced by bivalent IRV and induced by G1 type monovalent vaccines ; but there were significantly increase for the levels of serum IgG (t =2.691,P＜0.05) and serum neutralized antibody (t =2.561,P＜0.05) between induced by bivalent IRV and induced by G3 monovalent vaccines,there were no significant differences for other antibodies between induced by bivalent IRV and induced by G3 monovalent vaccines.At the same time,compared to negative control group,bivalent IRV induced significantly increase in the number of IFN-γ or IL-4 secreting cells in spleen lymphocytes.It was found that there were no significant differences for the number of IFN-γ or IL-4 secreting cells stimulated by G1 rotavirus between bivalent IRV and G1 monovalent vaccines; but there were significantly increase for the number of IL-4 secreting cells (t =2.327,P＜0.05) stimulated by G3 rotavirus between bivalent IRV and G3 monovalent vaccines,there were no significant differences for the number of IFN-γ secreting cells stimulated by G3 rotavirus between bivalent IRV and G3 type monovalent vaccines.Conclusion The bivalent IRV can induce effective immune response,in which there were no inhibitory interference between the components of bivalent IRV,which provided the experimental basis for the development of bivalent IRV.