Nunes I P, Jonsson R, Jensen H B, Bakken V
Broegelmann Research Laboratory for Microbiology, Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Bergen, Norway.
Oral Microbiol Immunol. 1996 Jun;11(3):142-9. doi: 10.1111/j.1399-302x.1996.tb00349.x.
The purpose of this study was to examine the antigenic abilities of Fusobacterium nucleatum strain ATCC 25586 and Porphyromonas gingivalis strain W50 black inbred BALB/cABom mice immunized subcutaneously. Furthermore, we aimed to analyze whether the outer membranes (OM) and whole cells (WC) of F. nucleatum or P. gingivalis had an effect on the levels of antibody response and whether a combination of both could either enhance or suppress the B-cell response. A single-cell assay, solid-phase enzyme-linked immunospot (ELISPOT), was used to analyze the splenic B-cell response (immunoglobulin A (IgA), IgG and IgM). Enzyme-linked immunosorbent assay (ELISA) and immunoblotting were used to verify the specific antibody response in the sera. A statistically significant lower level of spontaneous antibody production was observed in the group immunized with P. gingivalis OM compared with groups immunized with F. nucleatum and saline. The specific antibody titers measured by ELISA indicated that the bacterial preparations were able to induce IgG and IgM response. The preparations containing P. gingivalis OM induced higher humoral response than the preparations containing P. gingivalis WC, but for F. nucleatum such a difference was not observed. The prominent proteins revealed had apparent molecular masses of 40 kDa for F. nucleatum and 115, 55-56 and 43 kDa for P. gingivalis; whereas the immunoreactive proteins were 70, 65 and 40 kDa for mice immunized with F. nucleatum and 115, 55-56, 43 and 33-34 kDa for mice immunized with P. gingivalis. Quantitative analysis of B-cell response at the single cell level with ELISPOT revealed that some component(s) of P. gingivalis OM may have a suppressive ability on splenocytes incubated for a short time.
本研究旨在检测具核梭杆菌ATCC 25586菌株和牙龈卟啉单胞菌W50黑色近交系BALB/cABom小鼠皮下免疫后的抗原能力。此外,我们旨在分析具核梭杆菌或牙龈卟啉单胞菌的外膜(OM)和全细胞(WC)是否对抗体反应水平有影响,以及两者的组合是否会增强或抑制B细胞反应。采用单细胞检测方法——固相酶联免疫斑点(ELISPOT)分析脾脏B细胞反应(免疫球蛋白A(IgA)、IgG和IgM)。酶联免疫吸附测定(ELISA)和免疫印迹法用于验证血清中的特异性抗体反应。与用具核梭杆菌和生理盐水免疫的组相比,用牙龈卟啉单胞菌OM免疫的组中观察到自发抗体产生水平在统计学上显著降低。ELISA测定的特异性抗体滴度表明,细菌制剂能够诱导IgG和IgM反应。含有牙龈卟啉单胞菌OM的制剂比含有牙龈卟啉单胞菌WC的制剂诱导更高的体液反应,但对于具核梭杆菌未观察到这种差异。具核梭杆菌显示的突出蛋白质的表观分子量为40 kDa,牙龈卟啉单胞菌为115、55 - 56和43 kDa;而具核梭杆菌免疫的小鼠的免疫反应性蛋白质为70、65和40 kDa,牙龈卟啉单胞菌免疫的小鼠为115、55 - 56、43和33 - 34 kDa。用ELISPOT在单细胞水平对B细胞反应进行定量分析表明,牙龈卟啉单胞菌OM的某些成分可能对短时间孵育的脾细胞具有抑制能力。