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
6期
533-537
,共5页
闫立景%李剑%温婵%李嘉%蓝佳明%揣侠%高志云%张永红%金玉怀%王永祥
閆立景%李劍%溫嬋%李嘉%藍佳明%揣俠%高誌雲%張永紅%金玉懷%王永祥
염립경%리검%온선%리가%람가명%췌협%고지운%장영홍%금옥부%왕영상
柯萨奇病毒B3%腺病毒载体疫苗%DNA疫苗%Prime-boost免疫策略
柯薩奇病毒B3%腺病毒載體疫苗%DNA疫苗%Prime-boost免疫策略
가살기병독B3%선병독재체역묘%DNA역묘%Prime-boost면역책략
Coxsackievirus B3%Adenovirus vector vaccine%DNA vaccine%Prime-boost immuni-zation strategy
目的 构建重组腺病毒Ad/MDC-VP1,观察Ad/MDC-VP1与NA疫苗联合应用的免疫效果.方法 构建、包装重组腺病毒Ad/MDC-VP1并检测目的 蛋白的表达.BALB/c小鼠随机分为Ad/MDC-VP1、pcDNA3/MDC-VP1、pcDNA3/MDC-VPl+Ad/MDC-VP1和PBS 4组,肌肉注射免疫小鼠.用ELISA法和微量中和试验法分别检测血清柯萨奇病毒B3(CVB3)VP1 IgG和中和抗体滴度;CCK-8法检测淋巴细胞增殖活性和特异性CTL杀伤活性;用致死量CVB3攻击小鼠后,检测血中病毒滴度并观察小鼠的存活率.结果 成功构建并包装了重组腺病毒Ad/MDC-VP1,检测到目的 蛋白的表达.peDNA3/MDC-VP1+Ad/MDC-VP1组血清CVB3 VP1 IsG滴度、淋巴细胞增殖指数、CTL杀伤活性和对小鼠的保护率明显高于其他各组(P<0.05),血清病毒滴度低于其他各组(P<0.05).结论 Ad/MDC-VP1与DNA疫苗联合应用能显著提高小鼠细胞和体液免疫应答.
目的 構建重組腺病毒Ad/MDC-VP1,觀察Ad/MDC-VP1與NA疫苗聯閤應用的免疫效果.方法 構建、包裝重組腺病毒Ad/MDC-VP1併檢測目的 蛋白的錶達.BALB/c小鼠隨機分為Ad/MDC-VP1、pcDNA3/MDC-VP1、pcDNA3/MDC-VPl+Ad/MDC-VP1和PBS 4組,肌肉註射免疫小鼠.用ELISA法和微量中和試驗法分彆檢測血清柯薩奇病毒B3(CVB3)VP1 IgG和中和抗體滴度;CCK-8法檢測淋巴細胞增殖活性和特異性CTL殺傷活性;用緻死量CVB3攻擊小鼠後,檢測血中病毒滴度併觀察小鼠的存活率.結果 成功構建併包裝瞭重組腺病毒Ad/MDC-VP1,檢測到目的 蛋白的錶達.peDNA3/MDC-VP1+Ad/MDC-VP1組血清CVB3 VP1 IsG滴度、淋巴細胞增殖指數、CTL殺傷活性和對小鼠的保護率明顯高于其他各組(P<0.05),血清病毒滴度低于其他各組(P<0.05).結論 Ad/MDC-VP1與DNA疫苗聯閤應用能顯著提高小鼠細胞和體液免疫應答.
목적 구건중조선병독Ad/MDC-VP1,관찰Ad/MDC-VP1여NA역묘연합응용적면역효과.방법 구건、포장중조선병독Ad/MDC-VP1병검측목적 단백적표체.BALB/c소서수궤분위Ad/MDC-VP1、pcDNA3/MDC-VP1、pcDNA3/MDC-VPl+Ad/MDC-VP1화PBS 4조,기육주사면역소서.용ELISA법화미량중화시험법분별검측혈청가살기병독B3(CVB3)VP1 IgG화중화항체적도;CCK-8법검측림파세포증식활성화특이성CTL살상활성;용치사량CVB3공격소서후,검측혈중병독적도병관찰소서적존활솔.결과 성공구건병포장료중조선병독Ad/MDC-VP1,검측도목적 단백적표체.peDNA3/MDC-VP1+Ad/MDC-VP1조혈청CVB3 VP1 IsG적도、림파세포증식지수、CTL살상활성화대소서적보호솔명현고우기타각조(P<0.05),혈청병독적도저우기타각조(P<0.05).결론 Ad/MDC-VP1여DNA역묘연합응용능현저제고소서세포화체액면역응답.
Objective To construct recombinant adenovirus Ad/MDC-VP1 and investigate its im-muno-boosting effect of the mice primed with the experimental DNA vaccine against Coxsackievirus infection. Methods The recombinant adenovirus Ad/MDC-VP1 was constructed and packaged. The Western blot analysis was used to verify the target protein. BALB/c mice were divided into four groups: Ad/MDC-VP1 group, pcDNA3/MDC-VP1 group, pcDNA3/MDC-VP1 prime-Ad/MDC-VP1 boost group and PBS group. The mice in each group were immunized intramuscularly. The titers of serum IgG and neutralizing antibody were tested by ELISA and trace neutralization assay, respectively. The lymphocytes proliferation activity and specific CTL cytotoxic activity were tested by CCK-8 assay. The mice in each group were challenged with le-thal dose of Coxsackievirus, and the assay of the serum virus titers and the observation of protection efficacy against Coxsackievirus infection were carried out. Results The recombinant adenovirus Ad/MDC-VP1 was successfully constructed and the target protein was expressed. It was observed that the titers of CVB3 VP1 specific antibody, lymphocyte stimulation index, CTL cytotoxicity activities and protection rate of the pcDNA3/MDC-VP1 prime-Ad/MDC-VP1 boost group were much higher than those of the rest groups( P < 0.05), and the titer of serum virus was lower after CVB3 challenged ( P < 0.05 ). Conclusion Both the cellular and humoral immune responses in mice could been significantly enhanced by the pcDNA3/MDC-VP1 prime-Ad/MDC-VP1 boost strategy.
