The analysis of the immune response to the virulence factors of the periodontopathic bacteria

我们启动了一项研究，以确定RgpA和Kgp是否具有调理剂靶位点，以及这些位点在完整的牙龈卟啉单胞菌细胞上是否可用和可接近。免疫家兔，并从免疫前和免疫血清中分离IgG组分。通过ELISA测定抗体滴度，产生蛋白质印迹，并测量抗体对牙龈卟啉单胞菌调理作用的增强、吞噬作用以及代表四种血清型的牙龈卟啉单胞菌完整细胞的多形核白细胞的杀伤作用，来评估抗体的功能特征。RgpA和Kgp均诱导高滴度的IgG抗体。抗RgpA和抗Kgp抗体结合RgpA和Kgp两者，证明大比例的共享抗原表位。与免疫前IgG相比，这两种抗体与所有四种牙龈卟啉单胞菌血清型结合得同样好。两种抗体与RgpA和Kgp以及与四种牙龈卟啉单胞菌血清型的超声产物结合的免疫印迹模式几乎相同。这两种蛋白质诱导的抗体，显着增强调理作用，通过化学发光评估。两种抗体均显著增强PMN介导的对四种牙龈卟啉单胞菌血清型的细菌杀伤。这些数据表明这两种蛋白质似乎是抗牙龈卟啉单胞菌疫苗的潜在候选抗原。我们在RgpB上有类似的数据，它只具有催化结构域。提示RgpB也是抗牙龈卟啉单胞菌疫苗的潜在候选抗原

We initiated a study to determine whether RgpA and Kgp have opsonic target sites and whether these sites are available and accessible on intact P.gingivalis cells. Rabbits were immunized and IgG fractions were isolated from preimmune and immune sera. Functional characteristics of the antibodies were assessed by determining antibody titers by ELISA, generating Western blots, and measuring of the antibody enhancement of P.gingivalis opsonization, phagocytosis and killing by polymorphonuclear leukocytes of intact cells of P.gingivalis representative of the four serotypes. Both RgpA and Kgp induced high titers of IgG antibody. Anti-RgpA and anti-Kgp antibody bound to both RgpA and Kgp demonstrating a large proportion of shared antigenic epitopes. The two antibodies bound equally well to all four P.gingivalis serotypes compared to preimmune IgG. The immunoblot patterns of binding of the two antibodies to RgpA and Kgp and to sonlcates of the four P.gingivalis serotypes were virtually identical. Both proteins induced antibodies that significantly enhanced opsonization as assessed by chemiluminescence. Both antibodies significantly enhanced PMN-mediated bacterial killing of the four P.gingivalis serotypes. These data indicate these two proteins appear to be potential candidate antigens for an anti-P.gingivalis vaccine. We have similar data on RgpB, which possesses only a catalytic domain. The data indicate that RgpB also appear to be potential candidate antigen for anti-P.gingivalis vaccine