中华结核和呼吸杂志
中華結覈和呼吸雜誌
중화결핵화호흡잡지
Chinese Journal of Tuberculosis and Respiratory Diseases
2009年
7期
513-516
,共4页
宋楠楠%王秉翔%史大中%傅林锋%雒彧%于红娟%韩少波%焦磊%郄亚卿%王洪海%张颖%祝秉东
宋楠楠%王秉翔%史大中%傅林鋒%雒彧%于紅娟%韓少波%焦磊%郄亞卿%王洪海%張穎%祝秉東
송남남%왕병상%사대중%부림봉%락욱%우홍연%한소파%초뢰%극아경%왕홍해%장영%축병동
分枝杆菌,结核%疫苗,亚单位%卡介苗
分枝桿菌,結覈%疫苗,亞單位%卡介苗
분지간균,결핵%역묘,아단위%잡개묘
Mycobacterium tuberculosis%Vaccine subunit%BCG vaccine
目的 探讨二甲基三十六烷基铵(DDA)、卡介苗多糖核酸(BCG-PSN)作为结核病融合蛋白疫苗佐剂的免疫效应.方法 采用热酚法制备BCG-PSN,与DDA联合作为融合蛋白AMM(Ag85B-MPT64190-198-Mth8/4)佐剂免疫小鼠,设卡介苗、生理盐水、弗氏不完全佐剂(IFA)及单独应用DDA或BCG-PSN一种佐剂的各组作为对照.应用ELISA和酶联免疫斑点法检测免疫小鼠的体液与细胞免疫反应.结果 分离培养疫苗免疫小鼠的脾脏淋巴细胞,用特异性抗原AgSSB刺激后,AMM+DDA+BCG-PSN疫苗组、AMM+DDA组和卡介苗组小鼠每1×106个脾淋巴细胞分泌γ-干扰素的细胞数分别为222±79、259±85和230±64,均明显高于AMM+PBS组(40±4)、AMM+IFA组(10 ±3)、AMM+BCG-PSN组(132±18)和生理盐水组(8±4),差异有统计学意义(t值为2.923～7.118,均P<0.05);与单用DDA组比较,添加DDA和BCG-PSN两种佐剂的疫苗组小鼠血清抗体滴度Ig2a/IgG1较高(0.125和0.025),但γ-干扰素分泌水平没有增高.结论 AMM+DDA+BCG-PSN结核病亚单位疫苗能够引起较强的Th1细胞免疫为主的免疫反应,DDA+BCG-PSN(尤其是DDA)具有增强结核病亚单位疫苗Th1细胞免疫应答的佐剂效应.
目的 探討二甲基三十六烷基銨(DDA)、卡介苗多糖覈痠(BCG-PSN)作為結覈病融閤蛋白疫苗佐劑的免疫效應.方法 採用熱酚法製備BCG-PSN,與DDA聯閤作為融閤蛋白AMM(Ag85B-MPT64190-198-Mth8/4)佐劑免疫小鼠,設卡介苗、生理鹽水、弗氏不完全佐劑(IFA)及單獨應用DDA或BCG-PSN一種佐劑的各組作為對照.應用ELISA和酶聯免疫斑點法檢測免疫小鼠的體液與細胞免疫反應.結果 分離培養疫苗免疫小鼠的脾髒淋巴細胞,用特異性抗原AgSSB刺激後,AMM+DDA+BCG-PSN疫苗組、AMM+DDA組和卡介苗組小鼠每1×106箇脾淋巴細胞分泌γ-榦擾素的細胞數分彆為222±79、259±85和230±64,均明顯高于AMM+PBS組(40±4)、AMM+IFA組(10 ±3)、AMM+BCG-PSN組(132±18)和生理鹽水組(8±4),差異有統計學意義(t值為2.923～7.118,均P<0.05);與單用DDA組比較,添加DDA和BCG-PSN兩種佐劑的疫苗組小鼠血清抗體滴度Ig2a/IgG1較高(0.125和0.025),但γ-榦擾素分泌水平沒有增高.結論 AMM+DDA+BCG-PSN結覈病亞單位疫苗能夠引起較彊的Th1細胞免疫為主的免疫反應,DDA+BCG-PSN(尤其是DDA)具有增彊結覈病亞單位疫苗Th1細胞免疫應答的佐劑效應.
목적 탐토이갑기삼십륙완기안(DDA)、잡개묘다당핵산(BCG-PSN)작위결핵병융합단백역묘좌제적면역효응.방법 채용열분법제비BCG-PSN,여DDA연합작위융합단백AMM(Ag85B-MPT64190-198-Mth8/4)좌제면역소서,설잡개묘、생리염수、불씨불완전좌제(IFA)급단독응용DDA혹BCG-PSN일충좌제적각조작위대조.응용ELISA화매련면역반점법검측면역소서적체액여세포면역반응.결과 분리배양역묘면역소서적비장림파세포,용특이성항원AgSSB자격후,AMM+DDA+BCG-PSN역묘조、AMM+DDA조화잡개묘조소서매1×106개비림파세포분비γ-간우소적세포수분별위222±79、259±85화230±64,균명현고우AMM+PBS조(40±4)、AMM+IFA조(10 ±3)、AMM+BCG-PSN조(132±18)화생리염수조(8±4),차이유통계학의의(t치위2.923～7.118,균P<0.05);여단용DDA조비교,첨가DDA화BCG-PSN량충좌제적역묘조소서혈청항체적도Ig2a/IgG1교고(0.125화0.025),단γ-간우소분비수평몰유증고.결론 AMM+DDA+BCG-PSN결핵병아단위역묘능구인기교강적Th1세포면역위주적면역반응,DDA+BCG-PSN(우기시DDA)구유증강결핵병아단위역묘Th1세포면역응답적좌제효응.
Objective To investigate the activity of a novel adjuvant consisting of dimethyl-dioctyldecyl ammonium bromide (DDA) and BCG polysaccharide nucleic acid (BCG-PSN). Methods BCG-PSN was extracted by hot-phenol method, and combined with DDA and Mycobacterium tuberculosis fusion antigen AMM (Ag85B-MPT64190-198-Mtb8.4) to formulate the Mycobacterium tuberculosis subunit vaccine. Mice were iannunized subcutaneously with a 2-week interval between the immunizations (0. 2 ml/ dose), and humoral and cell-mediated immunity were detected by ELISA and ELISPOT respectively. Results With the stimulation of Ag85B in vitro, the number of antigen specific IFN-γ producing spleen lymphocytes were 222 ± 79,259 ± 85,230 ± 64 per million respectively in the mice immunized with AMM + DDA + BCG-PSN, AMM + DDA, and BCG. Spleen lymphocytes in these 3 groups produced higher levels of IFN-γ compared to the groups with the adjuvant of IFA or BCG-PSN alone or without adjuvant upon stimulation with Ag85B (t = 2.923 - 7. 118, P < 0.05). Furthermore, the adjuvant consisting of DDA and BCG-PSN increased the ratio of Ig2a/IgG1 than DDA alone (0. 125 vs 0. 025). Combined with AMM, the adjuvant DDA and the one consisting of DDA and BCG-PSN induced higher level of immunity than incomplete Freund' s adjuvant (IFA), NaCl, and BCG-PSN alone. Conclusion Mycobacterium tuberculosis subunit vaccine AMM + DDA + BCG-PSN induced a strong Thl-type immune response, and DDA + BCG-PSN, especially DDA promoted the immune response of the M. tuberculosis subunit vaccine in mice.
