Gemmell E, Sernia C, Grieco D A, Bird P S, Allen C J, Seymour G J
Immunopathology Laboratory, Oral Biology and Pathology, School of Dentistry, University of Queensland, Brisbane, Australia.
Oral Microbiol Immunol. 2001 Jun;16(3):129-35. doi: 10.1034/j.1399-302x.2001.016003129.x.
T-cell cytokine profiles, anti-Porphyromonas gingivalis antibodies and Western blot analysis of antibody responses were examined in BALB/c, CBA/CaH, C57BL6 and DBA/2J mice immunized intraperitoneally with different doses of P. gingivalis outer membrane antigens. Splenic CD4 and CD8 cells were examined for intracytoplasmic interleukin (IL)-4, interferon (IFN)-gamma and IL-10 by FACS analysis and levels of anti-P. gingivalis antibodies in the serum samples determined by enzyme-linked immunosorbent assay. Western blot analysis was performed on the sera from mice immunized with 100 microg of P. gingivalis antigens. The four strains of mice demonstrated varying degrees of T-cell immunity, although the T-cell cytokine profiles exhibited by each strain were not affected by different immunizing doses. While BALB/c and DBA/2J mice exhibited responses that peaked at immunizing doses of 100-200 microg of P. gingivalis antigens, CBA/CaH and C57BL6 demonstrated weak T-cell responsiveness compared with control mice. Like the T-cell responses, serum antibody levels were not dose dependent. DBA/2J exhibited the lowest levels of anti-P. gingivalis antibodies followed by BALB/c with CBA/CaH and C57BL6 mice demonstrating the highest levels. Western blot analysis showed that there were differences in reactivity between the strains to a group of 13 antigens ranging in molecular weight from 15 to 43 kDa. Antibody responses to a number of these bands in BALB/c mice were of low density, whereas CBA/CaH and C57BL6 mice demonstrated high-density bands and DBA/2J mice showed medium to high responses. In conclusion, different immunizing doses of P. gingivalis outer membrane antigens had little effect on the T-cell cytokine responses and serum anti-P. gingivalis antibody levels. Western blot analysis, however, indicated that the four strains of mice exhibited different reactivity to some lower-molecular-weight antigens. Future studies are required to determine the significance of these differences, which may affect the outcome of P. gingivalis infection.
在腹腔注射不同剂量牙龈卟啉单胞菌外膜抗原免疫的BALB/c、CBA/CaH、C57BL6和DBA/2J小鼠中,检测了T细胞细胞因子谱、抗牙龈卟啉单胞菌抗体及抗体反应的蛋白质印迹分析。通过流式细胞术分析脾脏CD4和CD8细胞的胞内白细胞介素(IL)-4、干扰素(IFN)-γ和IL-10,并通过酶联免疫吸附测定法测定血清样本中抗牙龈卟啉单胞菌抗体的水平。对用100μg牙龈卟啉单胞菌抗原免疫的小鼠血清进行蛋白质印迹分析。这四株小鼠表现出不同程度的T细胞免疫,尽管各株小鼠的T细胞细胞因子谱不受不同免疫剂量的影响。虽然BALB/c和DBA/2J小鼠在免疫剂量为100 - 200μg牙龈卟啉单胞菌抗原时反应达到峰值,但与对照小鼠相比,CBA/CaH和C57BL6表现出较弱的T细胞反应性。与T细胞反应一样,血清抗体水平也不依赖剂量。DBA/2J小鼠的抗牙龈卟啉单胞菌抗体水平最低,其次是BALB/c,而CBA/CaH和C57BL6小鼠的抗体水平最高。蛋白质印迹分析表明,各株小鼠对一组分子量在15至43 kDa之间的13种抗原的反应性存在差异。BALB/c小鼠对其中一些条带的抗体反应密度较低,而CBA/CaH和C57BL6小鼠显示出高密度条带,DBA/2J小鼠则表现出中等到高反应。总之,不同免疫剂量的牙龈卟啉单胞菌外膜抗原对T细胞细胞因子反应和血清抗牙龈卟啉单胞菌抗体水平影响不大。然而,蛋白质印迹分析表明,这四株小鼠对一些低分子量抗原表现出不同的反应性。需要进一步的研究来确定这些差异的意义,这些差异可能会影响牙龈卟啉单胞菌感染的结果。