Gp96 rich lysate as a vaccine candidate against infection with salmonella typhimurium

ABSTRACT

Glycoprotein 96 is the primary chaperone of the endoplasmic reticulum. Immunization with it induced potent Cytotoxic T lymphocyte responses to intracellular bacteria. S. typhimurium is a facultative intracellular bacterium and acquired resistance against this bacterium mainly depends on activity of Cytotoxic T cells. This study aimed to evaluate the capacity of Glycoprotein 96 rich lysate as a vaccine candidate to induce a protective immune response in mice against a lethal dose challenge with Salmonella typhimurium. Mice were infected with S. typhimurium. Then their spleens and livers were harvested and homogenized and the protein content of whole crude lysate was enriched using ammonium sulfate precipitation. SDS-polyacrylamide gel electrophoresis transfer method was used for enrichment of the protein from crude sample. Immunoblotting was conducted to detect Glycoprotein 96. Isoelectric point was achieved through the use of isoelectric focusing. PBS and whole crude lysate [from uninfected and infected mice] were injected to mice of test group, mice of control-1 group and mice of control-2 group, respectively, on days 0 and 14. Twenty-one days after the last immunization, the LD50 and bacterial loads of livers and spleens were determined. Immunization with Glycoprotein 96 rich lysate isolated from livers and spleens of S. typhimurium-infected mice induced protection against infection by S. typhimurium. Also, the bacterial burden of livers and spleens in mice that received gp96 rich lysate significantly decreased when compared to that of mice in the control groups