Statistical analysis Student��s t-test for unpaired values was used to determine the levels of significance. Analysis of variance (ANOVA) and a post hoc test (Dunnett��s method ��1 and Student-Newman-Keuls test) were employed selleckchem Bicalutamide when pairwise multiple comparison procedures were necessary. Differences between means were considered significant at a P<.05. RESULTS The binding capacity of human normal IgG (Figure 1A) and autoantibodies of patients with CP (Figure 1B) incubated with fibroblast cells is shown. Control IgG presented low binding (1A: 48 �� 6) and IgG from patients with CP showed high binding in fibroblast cells (1B: 229 �� 18). Moreover, when ��1 synthetic peptide corresponding to the second extracellular loop of human ��1-AR were included in the reaction, the binding capacity of normal control IgG was not modified (2A: 43 �� 6), while the autoantibodies of patients with CP were inhibited (2B: 53 �� 5).
Figure 1 Representative flow cytometry analysis of binding IgG. Culture fibroblasts were incubated with A: normal human IgG (1×10?9 M) or B: CP IgG (1×10?9 M) alone (1). The same concentration of normal (2 A) or CP (2 B) IgGs preincubated during … Figure 2 shows the immune reactivity of sera from different groups against human gingival fibroblast membranes. The optical density (OD) values for sera from CP were significantly higher than that from healthy individuals (control). Figure 2 Immunoreactivity (ELISA) of anti-membrane human gingival fibroblast antibodies of individual sera from groups: () 25 patients with CP and () 20 healthy subjects, control.
Sera (1/50 dilution) was assayed on sensitized microplates with … To determine the molecular interaction between IgG and human ��1-AR, OD values for each of the 45 subjects studied against ��1 synthetic peptide as coating antigen are shown in the scatterogram (Figure 3A). The immune reactivity of sera from CP was significantly higher than that from control individuals (P<.001). The OD of sera from CP was always at least 3 SD from that of sera from healthy individuals (control). On the other hand, when unrelated peptide (M1 cholinoreceptor peptide) was used as a coating antigen, both the IgG from patients with CP and IgG from healthy controls gave negative results (Figure 3B). Figure 3 Scatterogram showing immune reactivity (ELISA) of sera IgG antibodies against the second extracellular loop of A: ��1-AR and B: M1 cholinoreceptor tested by ELISA.
Shown are the individual optical density (OD) values each sera samples (1/50 dilution) … Figure 4 shows the concentration-dependent increase in OD values with affinity-purfied anti-��1 peptide IgG compared with the non-antipeptide Cilengitide IgG fraction eluted from the column from CP patients when the ��1-AR synthetic peptide was used as the coating antigen. The OD values of the anti-��1 synthetic peptide IgG were always greater than 3 SD of those of the non-antipeptide IgG (Figure 4A).