Journal of Microbiology

Joon Haeng Rhee

4 Articles

Immune Response Induced by ppGpp-Defective Salmonella enterica serovar Gallinarum in Chickens: Sang-Ik Park , Jae-Ho Jeong , Hyon E. Choy , Joon Haeng Rhee , Hee-Sam Na , Tae-Hoon Lee , Moon Her , Kyoung-Oh Cho , Yeongjin Hong; J. Microbiol. 2010;48(5):674-681. Published online November 3, 2010; DOI: https://doi.org/10.1007/s12275-010-0179-6

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To protect chickens from typhoid caused by Salmonella enterica serovar Gallinarum (S. Gallinarum), the attenuated 9R strain has been used in the field as a vaccine. However, safety concerns have been raised because the mutations in 9R are undefined while its efficacy is still a question under debate. A global regulator, ppGpp, synthesized by RelA and SpoT, has been shown to induce various virulence genes in S. Gallinarum (Jeong et al., 2008). In this study, two mutant strains defective in ppGpp-synthesis were constructed in wild-type S. Gallinarum (∆ppGpp) and 9R strain (9R-∆ppGpp) backgrounds and tested as live vaccines in chickens. After oral inoculation, the LD50 values of ∆ppGpp and 9R-∆ppGpp were approximately 5×1010 colony forming unit (CFU) similarly as 9R strain, which was ~105-fold higher than that of the wildtype S. Gallinarum strain. Immunological analyses revealed immunization with either of the two attenuated ppGpp-defective strains induced significant antibody responses, the production of antibody-secreting B cells in blood, proliferation of CD4+ and CD8+ T cells in the spleen, and splenic expression of proinflammatory cytokines, such as IFN-γ and TGF-β4, at levels comparable to the 9R strain. Chickens immunized with the mutants (1×108 CFU) were 80% protected against oral challenge with 1×109 wild-type virulent bacteria (4,000-fold LD50 dose), similar to the level of protection achieved by 9R immunization. Based on these data, live attenuated ∆ppGpp-defective strains may serve as novel vaccines to control fowl typhoid in chickens.

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Mutating both relA and spoT of enteropathogenic Escherichia coli E2348/69 attenuates its virulence and induces interleukin 6 in vivo
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Roy Curtiss
Avian Diseases.2023;[Epub] CrossRef
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Evaluation of Safety and Protective Efficacy of a waaJ and spiC Double Deletion Korean Epidemic Strain of Salmonella enterica Serovar Gallinarum
Jun-Feng Zhang, Ke Shang, Bai Wei, Yea-Jin Lee, Jong-Yeol Park, Hyung-Kwan Jang, Se-Yeoun Cha, Min Kang
Frontiers in Veterinary Science.2021;[Epub] CrossRef
Survival of the Fittest: The Relationship of (p)ppGpp With Bacterial Virulence
Shivani Kundra, Cristina Colomer-Winter, José A. Lemos
Frontiers in Microbiology.2020;[Epub] CrossRef
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Characterization of the RelBbu Regulon in Borrelia burgdorferi Reveals Modulation of Glycerol Metabolism by (p)ppGpp
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Generation of a safety enhanced Salmonella Gallinarum ghost using antibiotic resistance free plasmid and its potential as an effective inactivated vaccine candidate against fowl typhoid
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A novel approach for the generation of Salmonella Gallinarum ghosts and evaluation of their vaccine potential using a prime-booster immunization strategy
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Leif E. Sander
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Characterization of a Novel Inactivated Salmonella enterica Serovar Enteritidis Vaccine Candidate Generated Using a Modified cI857/λ PR/GeneEExpression System
Chetan V. Jawale, Atul A. Chaudhari, Byung Woo Jeon, Rahul M. Nandre, John Hwa Lee, A. J. Bäumler
Infection and Immunity.2012; 80(4): 1502. CrossRef
Role of RelA and SpoT in Burkholderia pseudomallei Virulence and Immunity
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Construction of a Salmonella Gallinarum ghost as a novel inactivated vaccine candidate and its protective efficacy against fowl typhoid in chickens
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Immunological Responses Induced by asd and wzy/asd Mutant Strains of Salmonella enterica serovar Typhimurium in BALB/c Mice: Hong Hua Piao , Vo Thi Minh Tam , Hee Sam Na , Hyun Ju Kim , Phil Youl Ryu , Soo Young Kim , Joon Haeng Rhee , Hyon E. Choy , Suhng Wook Kim , Yeongjin Hong; J. Microbiol. 2010;48(4):486-495. Published online August 20, 2010; DOI: https://doi.org/10.1007/s12275-010-0023-z

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Abstract PDF

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 LD50 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 (1×1010 CFU) or i.p. (1×107 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 LD50 (5×106 CFU). Taken together, these data indicate that HTSaYA and SHJ2104 could be developed as live attenuated Salmonella vaccine candidates.

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Protective Effect of Polygoni Cuspidati Radix and Emodin on Vibrio vulnificus Cytotoxicity and Infection: Jong Ro Kim , Dool-Ri Oh , Mi Hye Cha , Byoung Sik Pyo , Joon Haeng Rhee , Hyon E. Choy , Won Keun Oh , Young Ran Kim; J. Microbiol. 2008;46(6):737-743. Published online December 24, 2008; DOI: https://doi.org/10.1007/s12275-008-0232-x

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Abstract PDF

Vibrio vulnificus, a good model organism of bacterial septicemia, causes fatal septicemia manifesting a fulminating course and a high mortality rate within days. In order to identify new natural substances preventing V. vulnificus infection, a plant library was screened for inhibiting cytotoxicity to host cells by using Trypan blue staining and LDH assay. We found that Polygoni Cuspidati Radix potently suppressed the acute death of HeLa and RAW264.7 cells in a dose dependent manner. Further studies revealed that Polygoni Cuspidati Radix inhibited V. vulnificus growth and survival in HI broth and seawater, respectively. We confirmed that Polygoni Cuspidati Radix contained high level of emodin by thin layer chromatography (TLC). Emodin showed direct antibacterial activity against V. vulnificus. In addition, emodin prevented the morphologic damages and acute death of HeLa cells caused from V. vulnificus. The safety of Polygoni Cuspidati Radix and emodin to host cells was confirmed by MTT assay. Polygoni Cuspidati Radix and emodin protected mice from V. vulnificus infection.

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Factors Influencing Preferential Utilization of RNA Polymerase Containing Sigma-38 in Stationary-Phase Gene Expression in Escherichia coli: Eun Young Kim , Min-Sang Shin , Joon Haeng Rhee , Hyon E. Choy; J. Microbiol. 2004;42(2):103-110.; DOI: https://doi.org/2037 [pii]

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Abstract PDF: In order to understand the molecular basis of selective expression of stationary-phase genes by RNA polymerase containing [sigma]^38 (E[sigma]^38) in Escherichia coli, we examined transcription from the stationaryphase promoters, katEP, bolAP, hdeABP, csgBAP, and mcbP, in vivo and in vitro. Although these promoters are preferentially recognized in vivo by E[sigma]^38, they are transcribed in vitro by both E[sigma]^38 and E[sigma]^70 containing the major exponential [sigma], [sigma]^70. In the presence of high concentrations of glutamate salts, however, only E[sigma]^38 was able to efficiently transcribe from these promoters, which supports the concept that the promoter selectivity of [sigma]^38 -containing RNA polymerase is observed only under specific reaction conditions. The examination of 6S RNA, which is encoded by the ssr1 gene in vivo, showed that it reduced E[sigma]^70 activity during the stationary phase, but this reduction of activity did not result in the elevation of E[sigma]^38 activity. Thus, the preferential expression of stationary-phase genes by E[sigma]^38 is unlikely the consequence of selective inhibition of E[sigma]^70 by 6S RNA.

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