Immunological Responses Induced by asd and wzy/asd Mutant Strains of Salmonella enterica serovar Typhimurium in BALB/c Mice
2010
Piao, Hong Hua, Chonnam National University Medical School, Gwangju, Republic of Korea | Tam, Vo Thi Minh, Chonnam National University Medical School, Gwangju, Republic of Korea | Na, H.S., Dankook University College of Medicine, Cheonan, Republic of Korea | Kim, H.J., Chonnam National University Medical School, Gwangju, Republic of Korea | Ryu, P.Y., Chonnam National University Medical School, Gwangju, Republic of Korea | Kim, S.Y., Chonnam National University Medical School, Gwangju, Republic of Korea | Rhee, J.H., Chonnam National University Medical School, Gwangju, Republic of Korea | Choy, H.E., Chonnam National University Medical School, Gwangju, Republic of Korea | Kim, S.W., Korea University, Seoul, Republic of Korea | Hong, Y.J., Chonnam National University Medical School, Gwangju, Republic of Korea
Attenuated bacteria have long been developed as vaccine candidates but can have some disadvantages, such as the potential for damage to immune organs due to insufficient clearance. To minimize these disadvantages, we generated Salmonella enterica serovar Typhimurium mutants SHJ2104 (asd::cm) and HTSaYA (wzy::km, asd::cm). The wzy gene codes for the O-antigen polymerase, which is involved in lipopolysaccharide (LPS) biosynthesis, and asd codes for aspartate ß-semialdehyde dehydrogenase, which participates in cell wall formation. The strains synthesized LPS with a short-chain length, and showed lower cytotoxicity and reduced intracellular proliferation in animal cells compared to wild-type bacteria. After oral infection, the mutants were cleared in immune tissues, including the Peyer's patch, mesenteric lymph node, and spleen, within 5 days. The LD∧50 of the mutants in Balb/c mice was estimated to be 106 higher than wild-type bacteria when administered either via an oral or i.p. route, indicating that the two strains are highly attenuated. To compare the immune response to and protective effects of the mutants against wild-type bacterial infection, we inoculated the mutants into mice via an oral (1×10∨10 CFU) or i.p. (1×10∨7 CFU) route once or twice at a two week interval. All immune responses, such as serum IgG and secretory IgA levels, cytokine production, and delayed hypersensitivity, were highly induced by two rounds of immunization. HTSaYA and SHJ2104 induced similar immune responses, and mice immunized with HTSaYA or SHJ2104 via an i.p. route were protected against wild-type Salmonella infection even at 100-fold of the LD∧50 (5×10∨6 CFU). Taken together, these data indicate that HTSaYA and SHJ2104 could be developed as live attenuated Salmonella vaccine candidates.
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