Fig 2: CD40 interacts with CD11a. Sorted cells were Mn2+ activated, or not, then immunoprecipitation was performed with anti-CD40 antibody 4F11. A, Western blot for CD11a was performed. B, fold change of band intensities in response to Mn2+ was analyzed, and significant differences were calculated by one-tailed t test (for CD11a) or two-tailed t test (for CD40). Significant p-values are shown in the figure. Data are representative of three different experiments.

Fig 3: KGYY15binding is partially blocked by antibodies to CD11a and CD11b as well as by recombinant CD11a/CD18, CD11b/CD18, and CD40 proteins, but not recombinant CD28 protein. NAMALWA cells were stained with KGYY15-FITC in the absence/presence of competition. A, competition with anti-CD11a, anti-CD11b, or recombinant CD11a/CD18 or CD11b/CD18 proteins. B, competition with recombinant CD40 protein. C, competition with recombinant CD28 protein. Data are represented as mean ± SEM. p-Values were calculated by two-tailed t test and are shown in the figure.

Fig 4: CD4 negative and Th40 cells alter CD40 and CD18 surface expression in response to Mn2+. PBMCs were stained for CD4, CD40, CD18, CD11a, and CD11b and analyzed by flow cytometry. In the lymphocyte gate, cells were gated on CD4-negative (CD4neg), CD4lo, or CD4hi, then the expression of CD40 (A) and CD18 (B) was analyzed. Tinted histogram, isotype; solid line, no activation; dotted line, Mn2+ activation. p-Values were calculated, in A, by two-tailed t test and, in B, by one-way ANOVA with Tukey's multiple comparisons test.

Fig 5: KGYY15interacts with CD11a, CD11b, and CD18. Sorted NOD and BALB/c cells were activated with Mn2+, or not, then immunoprecipitation was performed with KGYY15-conjugated microbeads. A, Western blots for CD11a (top panels), CD11b (second from top panels), CD18 (third from top panels), and CD40 (bottom panels) were performed. B and C, fold change of band intensities in response to Mn2+ was analyzed, and significant differences were calculated by one-tailed t test (for CD11a, CD11b, and CD18; except for BALB/c PBMC large CD11a band where two-tailed test was used) or two-tailed t test (for CD40). Significant p-values are shown in the figure. Data are representative of least three different experiments.