中华地方病学杂志
中華地方病學雜誌
중화지방병학잡지
Chinese Journal of Endemiology
2015年
10期
717-722
,共6页
猪带绦虫%重组Bb-TSO45W-4B-TSOL18疫苗%保护性免疫应答%仔猪
豬帶縚蟲%重組Bb-TSO45W-4B-TSOL18疫苗%保護性免疫應答%仔豬
저대조충%중조Bb-TSO45W-4B-TSOL18역묘%보호성면역응답%자저
Taenia solium%Recombinant Bb-TSO45W-4B-TSOL18 vaccine%Protective immune responses%Domestic pigs
目的 探讨猪带绦虫重组Bb-TSO45W-4B-TSOL18疫苗免疫仔猪后,对猪带绦虫卵攻击感染后的保护性免疫应答.方法 将20头40日龄健康仔猪按体质量(约15 kg)采用随机数字表法分为5组,每组4头.分别为重组Bb-TSO45W-4B-TSOL18疫苗灌胃组、重组Bb-TSO45W-4B疫苗灌胃组、重组Bb-TSOL18疫苗灌胃组、空载体对照组和双歧杆菌液体培养基(MRS)对照组.各疫苗的免疫剂量为1×1011菌落形成单位(CFU),间隔2周免疫1次,共免疫2次.于末次免疫仔猪4周后,各组仔猪用猪带绦虫卵攻击感染,攻击感染后3个月剖杀仔猪,分离猪囊尾蚴,计算减蚴率;前腔静脉取血,分离血清和制备外周血淋巴细胞(PBMC).酶联免疫吸附试验(EHSA法)检测血清免疫球蛋白(Ig)G、IgG1、IgG2a水平和PBMC培养上清液中白细胞介素(IL)-2、干扰素(IFN)-γ、IL-4及IL-10水平;四甲基偶氮唑盐比色法(MTT法)检测PBMC增殖情况.结果 猪带绦虫重组Bb-TSO45W-4B-TSOL18、重组Bb-TSO45W-4B、重组Bb-TSOL18疫苗灌胃组减蚴率分别为83.09％、71.36％和74.85％.仔猪血清IgG、IgG1和IgG2a水平组间比较差异均有统计学意义(F=132.348、106.336、596.091,P＜0.05).其中重组Bb-TSO45W-4B-TSOL18、重组Bb-TSO45W-4B、重组Bb-TSOL18疫苗灌胃组IgG、IgG2a水平[(366.81±3.84)、(334.94±11.65)、(333.52±11.09),(87.74±0.95)、(84.48±0.80)、(84.30±1.09)mg/L]均高于MRS对照组[(245.94±8.81)、(62.61±0.84)mg/L,P均＜0.05],IgG1水平[(26.55±1.06)、(33.24±1.92)、(32.60±1.94)mg/L]低于MRS对照组[(42.78±0.87)mg/L,P均＜0.05].仔猪PBMC培养上清原液中IL-2、IFN-γ、IL-4和IL-10水平组间比较差异均有统计学意义(F=139.522、1 053.102、769.097、962.298,P均＜0.05).其中重组Bb-TSO45W-4B-TSOL18、重组Bb-TSO45W-4B、重组Bb-TSOL18疫苗灌胃组IL-2、IFN-γ水平[(212.24±3.12)、(205.91±3.18)、(205.85±4.35),(28.42±0.28)、(25.56±0.28)、(25.71±0.35)ng/L]均高于MRS对照组[(174.19±2.14)、(17.69±0.28)ng/L,P均＜0.05],IL-4、IL-10水平[(40.45±036)、(41.38±0.70)、(41.52±0.19),(71.45±0.83)、(73.38±0.70)、(74.77±0.41)ng/L]均低于MRS对照组[(52.57±0.29)、(94.82±0.45) ng/L,P均＜0.05].仔猪PBMC增殖水平组间比较差异有统计学意义(F=56.318,P＜0.05).其中重组Bb-TSO45W-4B-TSOL18、重组Bb-TSO45W-4B、重组Bb-TSOL 18疫苗灌胃组PBMC增殖水平(0.543±0.074、0.481±0.028、0.530±0.053)均高于MRS对照组(0.242±0.053,P均＜0.05).结论 猪带绦虫重组Bb-TSO45W-4B-TSOL 18疫苗能够诱导仔猪产生一定的保护力,其中Th1型免疫应答在其诱导的保护性免疫机制中起重要作用.
目的 探討豬帶縚蟲重組Bb-TSO45W-4B-TSOL18疫苗免疫仔豬後,對豬帶縚蟲卵攻擊感染後的保護性免疫應答.方法 將20頭40日齡健康仔豬按體質量(約15 kg)採用隨機數字錶法分為5組,每組4頭.分彆為重組Bb-TSO45W-4B-TSOL18疫苗灌胃組、重組Bb-TSO45W-4B疫苗灌胃組、重組Bb-TSOL18疫苗灌胃組、空載體對照組和雙歧桿菌液體培養基(MRS)對照組.各疫苗的免疫劑量為1×1011菌落形成單位(CFU),間隔2週免疫1次,共免疫2次.于末次免疫仔豬4週後,各組仔豬用豬帶縚蟲卵攻擊感染,攻擊感染後3箇月剖殺仔豬,分離豬囊尾蚴,計算減蚴率;前腔靜脈取血,分離血清和製備外週血淋巴細胞(PBMC).酶聯免疫吸附試驗(EHSA法)檢測血清免疫毬蛋白(Ig)G、IgG1、IgG2a水平和PBMC培養上清液中白細胞介素(IL)-2、榦擾素(IFN)-γ、IL-4及IL-10水平;四甲基偶氮唑鹽比色法(MTT法)檢測PBMC增殖情況.結果 豬帶縚蟲重組Bb-TSO45W-4B-TSOL18、重組Bb-TSO45W-4B、重組Bb-TSOL18疫苗灌胃組減蚴率分彆為83.09％、71.36％和74.85％.仔豬血清IgG、IgG1和IgG2a水平組間比較差異均有統計學意義(F=132.348、106.336、596.091,P＜0.05).其中重組Bb-TSO45W-4B-TSOL18、重組Bb-TSO45W-4B、重組Bb-TSOL18疫苗灌胃組IgG、IgG2a水平[(366.81±3.84)、(334.94±11.65)、(333.52±11.09),(87.74±0.95)、(84.48±0.80)、(84.30±1.09)mg/L]均高于MRS對照組[(245.94±8.81)、(62.61±0.84)mg/L,P均＜0.05],IgG1水平[(26.55±1.06)、(33.24±1.92)、(32.60±1.94)mg/L]低于MRS對照組[(42.78±0.87)mg/L,P均＜0.05].仔豬PBMC培養上清原液中IL-2、IFN-γ、IL-4和IL-10水平組間比較差異均有統計學意義(F=139.522、1 053.102、769.097、962.298,P均＜0.05).其中重組Bb-TSO45W-4B-TSOL18、重組Bb-TSO45W-4B、重組Bb-TSOL18疫苗灌胃組IL-2、IFN-γ水平[(212.24±3.12)、(205.91±3.18)、(205.85±4.35),(28.42±0.28)、(25.56±0.28)、(25.71±0.35)ng/L]均高于MRS對照組[(174.19±2.14)、(17.69±0.28)ng/L,P均＜0.05],IL-4、IL-10水平[(40.45±036)、(41.38±0.70)、(41.52±0.19),(71.45±0.83)、(73.38±0.70)、(74.77±0.41)ng/L]均低于MRS對照組[(52.57±0.29)、(94.82±0.45) ng/L,P均＜0.05].仔豬PBMC增殖水平組間比較差異有統計學意義(F=56.318,P＜0.05).其中重組Bb-TSO45W-4B-TSOL18、重組Bb-TSO45W-4B、重組Bb-TSOL 18疫苗灌胃組PBMC增殖水平(0.543±0.074、0.481±0.028、0.530±0.053)均高于MRS對照組(0.242±0.053,P均＜0.05).結論 豬帶縚蟲重組Bb-TSO45W-4B-TSOL 18疫苗能夠誘導仔豬產生一定的保護力,其中Th1型免疫應答在其誘導的保護性免疫機製中起重要作用.
목적 탐토저대조충중조Bb-TSO45W-4B-TSOL18역묘면역자저후,대저대조충란공격감염후적보호성면역응답.방법 장20두40일령건강자저안체질량(약15 kg)채용수궤수자표법분위5조,매조4두.분별위중조Bb-TSO45W-4B-TSOL18역묘관위조、중조Bb-TSO45W-4B역묘관위조、중조Bb-TSOL18역묘관위조、공재체대조조화쌍기간균액체배양기(MRS)대조조.각역묘적면역제량위1×1011균락형성단위(CFU),간격2주면역1차,공면역2차.우말차면역자저4주후,각조자저용저대조충란공격감염,공격감염후3개월부살자저,분리저낭미유,계산감유솔;전강정맥취혈,분리혈청화제비외주혈림파세포(PBMC).매련면역흡부시험(EHSA법)검측혈청면역구단백(Ig)G、IgG1、IgG2a수평화PBMC배양상청액중백세포개소(IL)-2、간우소(IFN)-γ、IL-4급IL-10수평;사갑기우담서염비색법(MTT법)검측PBMC증식정황.결과 저대조충중조Bb-TSO45W-4B-TSOL18、중조Bb-TSO45W-4B、중조Bb-TSOL18역묘관위조감유솔분별위83.09％、71.36％화74.85％.자저혈청IgG、IgG1화IgG2a수평조간비교차이균유통계학의의(F=132.348、106.336、596.091,P＜0.05).기중중조Bb-TSO45W-4B-TSOL18、중조Bb-TSO45W-4B、중조Bb-TSOL18역묘관위조IgG、IgG2a수평[(366.81±3.84)、(334.94±11.65)、(333.52±11.09),(87.74±0.95)、(84.48±0.80)、(84.30±1.09)mg/L]균고우MRS대조조[(245.94±8.81)、(62.61±0.84)mg/L,P균＜0.05],IgG1수평[(26.55±1.06)、(33.24±1.92)、(32.60±1.94)mg/L]저우MRS대조조[(42.78±0.87)mg/L,P균＜0.05].자저PBMC배양상청원액중IL-2、IFN-γ、IL-4화IL-10수평조간비교차이균유통계학의의(F=139.522、1 053.102、769.097、962.298,P균＜0.05).기중중조Bb-TSO45W-4B-TSOL18、중조Bb-TSO45W-4B、중조Bb-TSOL18역묘관위조IL-2、IFN-γ수평[(212.24±3.12)、(205.91±3.18)、(205.85±4.35),(28.42±0.28)、(25.56±0.28)、(25.71±0.35)ng/L]균고우MRS대조조[(174.19±2.14)、(17.69±0.28)ng/L,P균＜0.05],IL-4、IL-10수평[(40.45±036)、(41.38±0.70)、(41.52±0.19),(71.45±0.83)、(73.38±0.70)、(74.77±0.41)ng/L]균저우MRS대조조[(52.57±0.29)、(94.82±0.45) ng/L,P균＜0.05].자저PBMC증식수평조간비교차이유통계학의의(F=56.318,P＜0.05).기중중조Bb-TSO45W-4B-TSOL18、중조Bb-TSO45W-4B、중조Bb-TSOL 18역묘관위조PBMC증식수평(0.543±0.074、0.481±0.028、0.530±0.053)균고우MRS대조조(0.242±0.053,P균＜0.05).결론 저대조충중조Bb-TSO45W-4B-TSOL 18역묘능구유도자저산생일정적보호력,기중Th1형면역응답재기유도적보호성면역궤제중기중요작용.
Objective To study the protective immune responses induced by recombinant Bifidobacterium (Bb)-TSO45W-4B-TSOL18 vaccine of Taenia solium (T.solium) in domestic pigs challenged with T.solium eggs.Methods Twenty healthy 40 days old domestic pigs were divided into five groups by random number table according to body weight (15 kg):rBb-TSO45W-4B-TSOL18 vaccine group,rBb-TSO45W-4B vaccine group,rBb-TSOL18 vaccine group,blank vector control group and MRS control group.The content of vaccine in each vaccine group was 1 × 1011 CFU.A total of two immunization times was conducted,once every two weeks.Pigs were challenged with T.solium eggs 4 weeks after the last immunization and killed 3 months after infection.The cysticercus was counted and the reduction of the cysticercus was calculated.Blood was collected to separate sera and prepare peripheral blood lymphocytes (PBMC).The levels of IgG,IgG1 and IgG2a in sera were determined by enzyme linked immunosorbent assay (ELISA).The level of PBMC proliferation was tested using methyltetrazolium (MTT) assay.The levels of interleukin (IL)-2,interferon (IFN)-γ,IL-4 and IL-10 in PBMC culture supernatant were detected using ELISA.Results The reduction of cysticercus was 83.09％,71.36％ and 74.85％ in rBb-TSO45W-4B-TSOL18,rBb-TSO45W-4B and rBb-TSOL18 vaccine groups,respectively.The differences of IgG,IgG1,IgG2a levels in sera between groups were statistically significant (F =132.348,106.336,596.091,all P ＜0.05).The levels of IgG and IgG2a in rBb-TSO45W-4B-TSOL18,rBb-TSO45W-4B and rBb-TSOL18 vaccine groups [(366.81 ± 3.84),(334.94 ± 11.65),(333.52 ± 11.09),(87.74 ± 0.95),(84.48 ± 0.80),(84.30 ± 1.09)mg/L] were higher than those of the MRS control group [(245.94 ± 8.81),(62.61 ± 0.84)mg/L,all P ＜0.05].The levels of IgG1 in rBb-TSO45W-4B-TSOL18,rBb-TSO45W-4B and rBb-TSOL18 vaccine groups [(26.55 ± 1.06),(33.24 ± 1.92),(32.60 ± 1.94)mg/L] were lower than those of the MRS control group [(42.78 ± 0.87)mg/L,all P ＜0.05].The differences of IL-2,IFN-γ,IL-4 and IL-10 levels in PBMC original culture supernatant between groups were statistically significant (F =139.522,1 053.102,769.097,962.298,all P ＜0.05).The levels of IL-2 and IFN-γ in rBb-TSO45W-4B-TSOL18,rBb-TSO45W-4B and rBb-TSOL18 vaccine groups [(212.24 ± 3.12),(205.91 ± 3.18),(205.85 ± 4.35),(28.42 ± 0.28),(25.56 ± 0.28),(25.71 ± 0.35)ng/L] were higher than those of the MRS control group [(174.19 ± 2.14),(17.69 ± 0.28)ng/L,all P ＜0.05],while the levels of IL-4 and IL-10 [(40.45 ± 0.36),(41.38 ± 0.70),(41.52 ± 0.19),(71.45 ± 0.83),(73.38 ± 0.70),(74.77 ± 0.41)rig/L] were lower than those of the MRS control group [(52.57 ± 0.29),(94.82 ± 0.45)ng/L,all P ＜0.05].The differences of PBMC proliferation levels between groups were statistically significant (F =56.318,P ＜0.05).The PBMC proliferation levels in rBb-TSO45W-4B-TSOL18,rBb-TSO45W-4B and rBb-TSOL18 vaccine groups (0.543 ± 0.074,0.481 ± 0.028,0.530 ± 0.053) were higher than those of the MRS control group (0.242 ± 0.053,all P ＜0.05).Conclusions Recombinant Bb-TSO45W-4B-TSOL18 vaccine of T.solium could induce certain protection in domestic pigs.Type Th1 immune response may play an important role in induction of protective immunity.
