Gemmell E, Bird P S, Ford P J, Ashman R B, Gosling P, Hu Y, Seymour G J
Oral Biology and Pathology, School of Dentistry, The University of Queensland, Brisbane, Australia.
Oral Microbiol Immunol. 2004 Aug;19(4):247-51. doi: 10.1111/j.1399-302X.2004.00147.x.
Successive immunization of mice with Fusobacterium nucleatum and Porphyromonas gingivalis has been shown to modulate the specific serum IgG responses to these organisms. The aim of this study was to investigate these antibody responses further by examining the IgG subclasses induced as well as the opsonizing properties of the specific antibodies. Serum samples from BALB/c mice immunized with F. nucleatum (gp1-F), P. gingivalis (gp2-P), P. gingivalis followed by F. nucleatum (gp3-PF) F. nucleatum followed by P. gingivalis (gp4-FP) or saline alone (gp5-S) were examined for specific IgG1 (Th2) and IgG2a (Th1) antibody levels using an ELISA and the opsonizing properties measured using a neutrophil chemiluminescence assay. While IgG1 and IgG2a subclasses were induced in all immunized groups, there was a tendency towards an IgG1 response in mice immunized with P. gingivalis alone, while immunization with F. nucleatum followed by P. gingivalis induced significantly higher anti-P. gingivalis IgG2a levels than IgG1. The maximum light output due to neutrophil phagocytosis of P. gingivalis occurred at 10 min using nonopsonized bacteria. Chemiluminescence was reduced using serum-opsonized P. gingivalis and, in particular, sera from P. gingivalis-immunized mice (gp2-P), with maximum responses occurring at 40 min. In contrast, phagocytosis of immune serum-opsonized F. nucleatum demonstrated peak light output at 10 min, while that of F. nucleatum opsonized with sera from saline injected mice (gp5-S) and control nonopsonized bacteria showed peak responses at 40 min. The lowest phagocytic response occurred using gp4-FP serum-opsonized F. nucleatum. In conclusion, the results of the present study have demonstrated a systemic Th1/Th2 response in mice immunized with P. gingivalis and/or F. nucleatum with a trend towards a Th2 response in P. gingivalis-immunized mice and a significantly increased anti-P. gingivalis IgG2a (Th1) response in mice immunized with F. nucleatum prior to P. gingivalis. Further, the inhibition of neutrophil phagocytosis of immune serum-opsonized P. gingivalis was modulated by the presence of anti-F. nucleatum antibodies, while anti-P. gingivalis antibodies induced an inhibitory effect on the phagocytic response to F. nucleatum.
已证明用具核梭杆菌和牙龈卟啉单胞菌对小鼠进行连续免疫可调节针对这些微生物的特异性血清IgG反应。本研究的目的是通过检查诱导产生的IgG亚类以及特异性抗体的调理特性来进一步研究这些抗体反应。使用酶联免疫吸附测定(ELISA)检测用具核梭杆菌(gp1-F)、牙龈卟啉单胞菌(gp2-P)、先牙龈卟啉单胞菌后具核梭杆菌(gp3-PF)、先具核梭杆菌后牙龈卟啉单胞菌(gp4-FP)或单独用生理盐水(gp5-S)免疫的BALB/c小鼠血清样本中的特异性IgG1(Th2)和IgG2a(Th1)抗体水平,并使用中性粒细胞化学发光测定法测量调理特性。虽然在所有免疫组中均诱导出IgG1和IgG2a亚类,但单独用牙龈卟啉单胞菌免疫的小鼠有产生IgG1反应的趋势,而先用具核梭杆菌免疫后再用牙龈卟啉单胞菌免疫诱导出的抗牙龈卟啉单胞菌IgG2a水平明显高于IgG1。对于未调理的细菌,牙龈卟啉单胞菌被中性粒细胞吞噬导致的最大光输出在10分钟时出现。使用血清调理的牙龈卟啉单胞菌时化学发光降低,尤其是来自牙龈卟啉单胞菌免疫小鼠(gp2-P)的血清,最大反应在40分钟时出现。相比之下,免疫血清调理的具核梭杆菌吞噬的峰值光输出在10分钟时出现,而用注射生理盐水小鼠(gp5-S)的血清调理的具核梭杆菌和对照未调理细菌的峰值反应在40分钟时出现。使用gp4-FP血清调理的具核梭杆菌时吞噬反应最低。总之,本研究结果表明,在用牙龈卟啉单胞菌和/或具核梭杆菌免疫的小鼠中存在全身性Th1/Th2反应,在牙龈卟啉单胞菌免疫的小鼠中有Th2反应的趋势,而在先用具核梭杆菌免疫后再用牙龈卟啉单胞菌免疫的小鼠中抗牙龈卟啉单胞菌IgG2a(Th1)反应显著增加。此外,抗具核梭杆菌抗体的存在调节了免疫血清调理血清调理的牙龈卟啉单胞菌中性粒细胞吞噬的抑制作用,而抗牙龈卟啉单胞菌抗体对具核梭杆菌的吞噬反应诱导了抑制作用。