Recombinant Ax21 protein is a promising subunit vaccine candidate against Stenotrophomonas maltophilia in a murine infection model.

Stenotrophomonas maltophilia is an emerging pathogen that can cause several disease manifestations such as bacteremia, meningitis, respiratory tract infections and others. More seriously, this pathogen has a highly evolving antibiotic resistance profile. Antibiotic misuse is further aggravating the situation by inducing the development of multi- and even pan-resistance. Thus, employing diverse strategies to overcome this increasing antibiotic resistance is of paramount importance. In general, vaccination is one of these strategies that prevents the onset of infection, provides long term protection against infection, and most importantly diminishes the antibiotic consumption, thus, resulting in controlling resistance. Unfortunately, vaccine research concerning S. maltophilia is very scarce in the literature. Ax21 protein is an outer membrane protein implicated in several virulence mechanisms of S. maltophilia such as quorum sensing, biofilm formation, and antibiotic resistance. Our computational analysis of Ax21 revealed its potential immunogenicity. In the current study, Ax21 protein of S. maltophilia was cloned and heterologously expressed in Escherichia coli. Mice were immunized with the purified recombinant antigen using Bacillus Calmette-Guérin(BCG) and incomplete Freund's adjuvant (IFA) as immune-adjuvants. Enzyme-linked immunosorbent assay (ELISA) revealed significant antigen-specific IgG1, IgG2a and total IgG levels in immunized mice which reflected successful immune stimulation. Immunized mice that were challenged with S. maltophilia showed a substantialreduction in bacterial bioburden in lungs, liver, kidneys, and heart. In addition, liver histological examination demonstrated a remarkable decrease in pathological signs such as necrosis, vacuolation, bile duct fibrosis and necrosis, infiltration of inflammatory cells, and hemorrhage. Whole cell ELISA and opsonophagocytic assay confirmed the ability of serum antibodies from immunized mice to bind and facilitate phagocytosis of S. maltophilia, respectively. To our knowledge, this is the first report to demonstrate the vaccine protective efficacy of Ax21 outer membrane protein against S. maltophilia infection.