Comparative Analysis of Immune Responses to Outer Membrane Antigens OMP10, OMP19, and OMP28 of Brucella abortus

Abstract

Brucella infection is accompanied by cytokine production, which serves as an important factor to evaluate the innate and adaptive immune responses. Several researchers have been investigating the mechanisms involved in Brucella infection in the host. Here, we conducted an analytical study to define pathogenic pathways and immune mechanisms involved in Brucella infection by investigating the antigenic efficacy of recombinant outer membrane protein 10 (rOMP10), outer membrane protein 19 (rOMP19), and outer membrane protein 28 (rOMP28) in vitro and in vivo upon stimulation/immunization. Cytokine production was analyzed by nitric oxide (NO) assay and enzyme-linked immunosorbent assay (ELISA) after stimulation of RAW 264.7 cells and naive splenocytes with the recombinant proteins. Our results show that levels of NO, tumor necrosis factor (TNF)-alpha, and interleukin (IL)-6 increased in RAW 264.7 cells in a time-dependent manner following recombinant protein stimulation. In contrast, levels of interferon (1FN)-gamma and IL-2 increased in naive splenocytes after stimulation with proteins. ELISA and ELISpot assays were performed after immunization of mice with recombinant proteins. rOMP28 greatly increased IFN-gamma, IL-2, and TNF-alpha levels than IL-4 and IL-6 levels in vitro. Of the recombinant proteins, rOMP19 elicited a mixed Thl/Th2 immune response by increasing the number of IgG-secreting cells in vivo.