ASSESSMENT OF PROTECTIVE EFFECT OF HEAT SHOCK PROTEIN-LYPOPOLYSACCHARIDE OF SALMONELLA TYPHIMURIUM RECOMBINANT CONSTRUCTION IN MICE

Aim. To study protective activity of recombinant construction of heat-shock protein with lypopolysaccharide (rcHSP-LPS) as well as its variants (with destroyed protein or bounded LPS) against Salmonella typhimurium . It was also planned to study the ability of rcHSP-LPS to interact with tolllike receptors (TLRs) expressed on continuous cell lines. Materials and methods. One of the following preparations was administered to outbred mice: rcHSP-LPS; rcHSP-LPS treated by polymyxin B (PMB) for bounding of LPS — rc(HSP-LPS)PMB; rcHSP-LPS in which protein was treated by boiling during 30 min — rc(HSP-LPS)B; LPS ( E.coli K-235); polymyxin B (PMB). Twenty-four hours after single or last administration of rcHSP-LPS, each mice was intraperitoneally inoculated with 63 LD 5 0 of S.typhimurium 415 contained in 0.5 ml of physiologic solution. Antibody titer to LPS of Salmonella typhimurium was measured by immunoenzyme assay. Results. It was demonstrated that rcHSP-LPS administered 24 hours before inoculation induced resistance to S.typhimurium infection. Protection formed after 3 injections of rcHSP-LPS with 10 mcg in each or single injection with 100 mcg/mouse. Forty to eighty percent of immunized mice survived after challenge while 90% of control animals died. Destroy of the HSP by boiling of the construction led to loss of protective effect. Bounding of LPS by PMB did not lead to loss of protective properties of the construction but they expressed only after its multiple administration with 10 mcg per mouse. LPS of E.coli in dose 0.0266 mcg per mouse as well as PMB did not influence the course of S.typhimurium infection in mice. Conclusion. It was shown that rcHSP-LPS effectively protects mice from S.typhimurium infection by activating innate immunity; one of the possible mechanisms for such protection determined by interaction with TLRs 2 and 4 was considered. Other studies are needed in order to elucidate other mechanisms of innate immunity, which can be activated by rcHSP-LPS.

S. typhimurium, rcHSP-LPS, HSP, S.typhimurium, protection, TLR2, TLR4