title: Outer membrane protein A (OmpA) of Shigella flexneri 2a induces TLR2-mediated activation of B cells: involvement of protein tyrosine kinase, ERK and NF-κB.
abstract: B cells are critically important in combating bacterial infections and their differentiation into plasma cells and memory cells aids bacterial clearance and long-lasting immunity conferred by essentially all vaccines. Outer membrane protein A (OmpA) of Shigella flexneri 2a has been demonstrated to induce the production of IgG and IgA in vivo following immunization of mice through intranasal route, but the direct involvement of B cells in OmpA-mediated immune regulation was not determined. Consequently, we investigated whether OmpA can modulate B cell functions and identified the molecular events involved in OmpA-induced B cell immune response in vitro. We show that OmpA of S. flexneri 2a activates B cells to produce protective cytokines, IL-6 and IL-10 as well as facilitates their differentiation into antibody secreting cells (ASCs). The immunostimulatory properties of OmpA are attributed to the increased surface expression of MHCII and CD86 on B cells. We also report here that B cell activation by OmpA is mediated strictly through recognition by TLR2, resulting in initiation of cascades of signal transduction events, involving increased phosphorylation of protein tyrosine kinases (PTKs), ERK and IκBα, leading to nuclear translocation of NF-κB. Importantly, a TLR2 antibody diminishes OmpA-induced upregulation of MHCII and CD86 on B cell surface as well as significantly inhibits B cell differentiation and cytokine secretion. Furthermore, we illustrate that B cell differentiation into ASCs and induction of cytokine secretion by OmpA are dependent on PTKs activity. Moreover, we identify that OmpA-induced B cell differentiation is entirely dependent on ERK pathway, whereas both NF-κB and ERK are essential for cytokine secretion by B cells. Overall, our data demonstrate that OmpA of S. flexneri 2a amplifies TLR signaling in B cells and triggers B cell immune response, which is critical for the development of an effective adaptive immunity to an optimal vaccine antigen.
authors.affiliation: Division of Pathophysiology, National Institute of Cholera and Enteric Diseases, Kolkata, West Bengal, India.
authors.forename: Rajsekhar
authors.initials: R
authors.lastname: Bhowmick
authors.affiliation: Division of Pathophysiology, National Institute of Cholera and Enteric Diseases, Kolkata, West Bengal, India.
authors.forename: Debasis
authors.initials: D
authors.lastname: Pore
authors.affiliation: Division of Pathophysiology, National Institute of Cholera and Enteric Diseases, Kolkata, West Bengal, India.
authors.forename: Manoj K
authors.initials: MK
authors.lastname: Chakrabarti
mesh_terms: D000818:Animals; D000921:Antibody-Producing Cells / Q000378:metabolism; D017951:Antigen Presentation / Q000187:drug effects; D001402:B-Lymphocytes / Q000166:cytology* / Q000187:drug effects; D051940:B7-2 Antigen / Q000378:metabolism; D001425:Bacterial Outer Membrane Proteins / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D057809:HEK293 Cells; D000949:Histocompatibility Antigens Class II / Q000378:metabolism; D006801:Humans; D016753:Interleukin-10 / Q000096:biosynthesis; D015850:Interleukin-6 / Q000096:biosynthesis; D008213:Lymphocyte Activation / Q000276:immunology; D020935:MAP Kinase Signaling System; D008807:Mice, Inbred BALB C; D016328:NF-kappa B / Q000378:metabolism*; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D012763:Shigella flexneri / Q000378:metabolism*; D051195:Toll-Like Receptor 2 / Q000235:genetics / Q000378:metabolism*; D051216:Toll-Like Receptor 6 / Q000378:metabolism; D015854:Up-Regulation / Q000235:genetics
chemical_list: D051940:B7-2 Antigen; D001425:Bacterial Outer Membrane Proteins; D000949:Histocompatibility Antigens Class II; D015850:Interleukin-6; D016328:NF-kappa B; D051195:Toll-Like Receptor 2; D051216:Toll-Like Receptor 6; D016753:Interleukin-10; C078167:OMPA outer membrane proteins; D011505:Protein-Tyrosine Kinases; D048049:Extracellular Signal-Regulated MAP Kinases