日本血吸虫重组两歧双歧杆菌pGEX—Sj14—3—3疫苗免疫BALB／c小鼠的血清抗体动态观察

Dynamic observation on levels of antibodies in sera of BALB/c mice immunized with recombinant Bifutobacterium bifidum pGEX-Sj14-3-3 vaccine of Schistosoma japonicum

目的动态观察重组两歧双歧杆菌（Bb）pGEX—Sj14—3—3疫苗免疫BALB／c小鼠后血清IgG及其亚类、IgE和IgA的变化。方法将96只BALB／c小鼠按体质量随机分为2组，每组48只．分别经口服灌胃（PO组）和鼻腔黏膜途径（IN组）用重组Bb（pGEX-Sj14—3—3）疫苗单次免疫小鼠，免疫后0、2、4、6、8、10、12、14、16、18、20、22周时每组各剖杀4只小鼠，眼球取血，采用常规酶联免疫吸附实验（ELISA法）检测小鼠血清IgG及其亚类、IgE和IgA水平。结果P0组和IN组小鼠血清IgG、IgG1、IgG2a、IgG2b、IgG3、IgE、IgA水平分别在免疫后2—22、2～14、2～22、2～22、2～20、2～22、2—22周升高，PO组分别在8、6、6、4、8、10、6周达最高水平（0．065±0．001、0．021±0．002、0．011±0．001、0．015±0．003、0．014±0．002、0．011±0．001、0．013±0．002），与同组0周（0．032±0．001、0．015±0．002、0．005±0.002、0．005±0．001、0．006±0．001、0．006±0．001、0．005±0．001）比较，除IgG1、IgG2b外，差异有统计学意义（P均〈0．05或〈0．01）；而IN组分别在4、6、4、4、8、10、8周达最高水平（0．064±0．003、0．022±0．002、0．012±0．003、0．019±0．001、0．013±0．001、0．015±0．001、0．01±-0．003），与同组0周（0．032±0．001、0．015±0．002、0．005±0．002、0．005±0．001、0．006±0．001、0．006±0．001、0．005±0．001）比较，除IgG1、IgA外，差异均有统计学意义（P均〈0．05或〈0．01）。结论重组Bb（pGEX—sj14—3—3）疫苗在免疫早期（2—10周）可通过促进IgG2a、IgG2b、IgG3和IgG1、IgE、IgA合成诱导小鼠产生Th1和Th2混合型免疫应答。

Objective To study the dynamic changes of IgG, IgG subclasses, IgE and IgA in sera of BALB/c mice immunized with recombinant Bifidobacterium bifidum （Bb） pGEX-Sj14-3-3 vaccine of Schistosoma japonicum. Methods Ninty six BALB/c mice were randomly divided into two groups： oral immunization group and intranasal immunization group, 48 mice in each group. Mice were orally and intranasally immunized with recombinant Bb（pGEX-Sj14-3-3） vaccine, .respectively. Four mice from each group were sacrificed, respectively, on weeks 0, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20 and 22 after immunization and their sera from the eyeballs were collected. The levels of IgG, IgG subclasses, IgE and IgA were assayed with routine Enzyme-linked immunosorbent assay（ELISA）. Results The titers of IgG, IgG1, IgG2a, IgG2b, IgG3, IgE and IgA in both groups increased during the 2 - 22th, 2 - 14th, 2 - 22th, 2 - 22th, 2 - 20th, 2 - 22th, 2 - 22th weeks, respectively. The values reached the highest level on weeks 8, 6, 6, 4, 8, 10 and 6, respectively, in the oral group, and the values were （0.065 ± 0.001, 0.021 ± 0.002, 0.011 ± 0.001, 0.015 ±0.003, 0.014 ± 0.002, 0.011 ± 0.001, 0.013 ± 0.002）, respectively, as compared with the values on week 0（0.032 ± 0.001, 0.015 ± 0.002, 0.005 ± 0.002, 0.005 ± 0.001, 0.006 ±0.001, 0.006 ± 0.001, 0.005 ± 0.001）, the differences were statistically significant（P 〈 0.05 or 〈 0.01） except that of IgG1 and IgG2b. In the intranasal group these values reached the highest levels on weeks 4, 6, 4, 4, 8, 10 and 8, respectively, and the values were （0.064 ± 0.003, 0.022 ± 0.002, 0.012 ± 0.005, 0.019 ± 0.001, 0.013 ± 0.001, 0.015 ± 0.001, 0.014 ± 0.003）, respectively, as compared with the value on week 0, the differences were statistically significant（P 〈0.05 or 〈 0.01） except that of IgG1 and IgA. Conclusions Types Thl and Th2 mixed type immune responses can be induced in mice by immunization with the recombinant Bb （pGEX-Sj14-3-3） vaccine by early period of immunization （2th - 10th week）.