中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2009年
7期
642-645
,共4页
曾瑞红%李广学%凌世淦%张贺秋%姚智燕%杨建岭%贺峰%黄睿%刘艳坤%魏林
曾瑞紅%李廣學%凌世淦%張賀鞦%姚智燕%楊建嶺%賀峰%黃睿%劉豔坤%魏林
증서홍%리엄학%릉세감%장하추%요지연%양건령%하봉%황예%류염곤%위림
丙型肝炎病毒%多表位抗原%联合免疫%免疫原性
丙型肝炎病毒%多錶位抗原%聯閤免疫%免疫原性
병형간염병독%다표위항원%연합면역%면역원성
HCV%Multi-epitope antigen%Co-immunization%Immunogenicity
目的 研究两个丙型肝炎病毒(HCV)多表位重组抗原联合免疫后诱导的细胞免疫和体液免疫应答,以及对小鼠的保护作用.方法 用两个多表位抗原HCV-T和HCV-E1联合免疫BALB/c小鼠3次,ELISA检测血清中抗体IgG、IgG1和IgG2a滴度;最后一次免疫后10 d杀死一半小鼠,取脾细胞,用乳酸脱氢酶(LDH)释放法检测细胞毒性T淋巴细胞(CTL)活性;ELISPOT法检测分泌IFN-γ和IL4的细胞;最后一次免疫后2周,在免疫小鼠的背部皮下注射106个SP2/0-NS3细胞,考察联合免疫对小鼠的保护作用;另取BALB/c小鼠,先在背部皮下注射106个SP2/0-NS3细胞,7 d后开始联合免疫,免疫3次,考察免疫治疗作用.结果 用HCV-T+HCV-E1联合免疫诱导了高滴度的HCV-E1特异性的IgG、IgG1和IgG2a抗体以及高水平的CTL活性;与单独免疫相比,联合免疫产生的分泌IFN-γ和IL-4的细胞数量有协同作用;而且联合免疫能有效预防和治疗SP2/0-NS3对小鼠的攻击.结论 HCV-T+HCV-E1联合免疫诱导了保护性的体液免疫和细胞免疫应答,有望进一步开发为一种有效的重组HCV疫苗.
目的 研究兩箇丙型肝炎病毒(HCV)多錶位重組抗原聯閤免疫後誘導的細胞免疫和體液免疫應答,以及對小鼠的保護作用.方法 用兩箇多錶位抗原HCV-T和HCV-E1聯閤免疫BALB/c小鼠3次,ELISA檢測血清中抗體IgG、IgG1和IgG2a滴度;最後一次免疫後10 d殺死一半小鼠,取脾細胞,用乳痠脫氫酶(LDH)釋放法檢測細胞毒性T淋巴細胞(CTL)活性;ELISPOT法檢測分泌IFN-γ和IL4的細胞;最後一次免疫後2週,在免疫小鼠的揹部皮下註射106箇SP2/0-NS3細胞,攷察聯閤免疫對小鼠的保護作用;另取BALB/c小鼠,先在揹部皮下註射106箇SP2/0-NS3細胞,7 d後開始聯閤免疫,免疫3次,攷察免疫治療作用.結果 用HCV-T+HCV-E1聯閤免疫誘導瞭高滴度的HCV-E1特異性的IgG、IgG1和IgG2a抗體以及高水平的CTL活性;與單獨免疫相比,聯閤免疫產生的分泌IFN-γ和IL-4的細胞數量有協同作用;而且聯閤免疫能有效預防和治療SP2/0-NS3對小鼠的攻擊.結論 HCV-T+HCV-E1聯閤免疫誘導瞭保護性的體液免疫和細胞免疫應答,有望進一步開髮為一種有效的重組HCV疫苗.
목적 연구량개병형간염병독(HCV)다표위중조항원연합면역후유도적세포면역화체액면역응답,이급대소서적보호작용.방법 용량개다표위항원HCV-T화HCV-E1연합면역BALB/c소서3차,ELISA검측혈청중항체IgG、IgG1화IgG2a적도;최후일차면역후10 d살사일반소서,취비세포,용유산탈경매(LDH)석방법검측세포독성T림파세포(CTL)활성;ELISPOT법검측분비IFN-γ화IL4적세포;최후일차면역후2주,재면역소서적배부피하주사106개SP2/0-NS3세포,고찰연합면역대소서적보호작용;령취BALB/c소서,선재배부피하주사106개SP2/0-NS3세포,7 d후개시연합면역,면역3차,고찰면역치료작용.결과 용HCV-T+HCV-E1연합면역유도료고적도적HCV-E1특이성적IgG、IgG1화IgG2a항체이급고수평적CTL활성;여단독면역상비,연합면역산생적분비IFN-γ화IL-4적세포수량유협동작용;이차연합면역능유효예방화치료SP2/0-NS3대소서적공격.결론 HCV-T+HCV-E1연합면역유도료보호성적체액면역화세포면역응답,유망진일보개발위일충유효적중조HCV역묘.
Objective To investigate the cellular and humoral immune responses and protective effect induced by co-immunization with two multi-epitope combinant antigens. Methods Mice were co-im-munized with the muhi-epitope HCV-T and HCV-E1 antigens three times. Sera antibodies IgG, IgG1 and IgG2a were tested by ELISA. Spleens from BALB/c mice immunized were removed 10 days after the last im-munization. CTL activity was assessed using LDH cytotoxicity assay kit. IFN-γ- and IL-4-secreting cells were quantified using ELISPOT kit. Two weeks after the final immunization, the mice were challenged sub-cutaneously(s, c. ) at the back with 106 SP2/0-NS3 cells, and protective effect was observed. For therapy, 106 SP2/0-NS3 cells were implanted into the back of BALB/c mice. Seven days later, mice were immuniza-tion three times. Therapy effect was observed. Results Co-immunization with HCV-T and HCV-E1 induced high tiers of HCV-El-specific IgG, IgG1 and IgG2a antibodies, and high level of CTL activity. Synergistic effect in frequencies of both specific IFN-γ-secreting cells and IL-4-secreting cells was observed in mice co-immunized. Prophylactic as well as therapeutic administration of mT + mE1 in mice led to protecting mice against SP2/0-NS3 cells. These results suggested that mT + mE1 was potential as a prophylactic as well as therapeutic HCV vaccine. Conclusion Co-immunization with HCV-T + HCV-EI induced protective humor-al and cellular immune response. HCV-T + HCV-E1 was potential as a recombinant HCV vaccine.
