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Invasion of Mammalian Cells by Rough Variant of Mycobacterium abscessus
Whang, Jake,Back, Young Woo,Lee, Gang-In,Kim, Hwa-Jung 대한미생물학회 2016 Journal of Bacteriology and Virology Vol.46 No.4
One of a rapid growing mycobacteria (RGM), Mycobacterium abscessus (MAB), is the most causative agents of RGM pulmonary disease. MAB can change their morphology that smooth (S) type to more virulent type of rough (R). Bacterial invasion into host cells is an important first step to initiate their infection. The phagocytic and invasion mechanisms of Mycobacterium tuberculosis through the host-parasite interaction have been researched. Although MAB causes a wide range of clinical diseases, little is known about their invasion ability or why the R type is more virulent than the S type. To compare invasion ability of R with S types, their infection abilities to dermal fibroblast, HaCaT cells, A549 cells and bone marrow derived macrophages were analyzed. After 2 h of infection, intracellular survival numbers of the R type were significantly higher in all infected cells than S types. The fluorescence-activated cell sorting (FACS) and confocal microscopy assay also revealed that red fluorescent amount and intracellular bacterial numbers in all of the cells infected with MAB R type expressing the red fluorescent protein (RFP) were significantly higher than the S type. Our data suggest that the virulence of MAB is proportionally related to the invasion ability into mammalian cells and macrophages.
( Yumi Park ),( Jae-gook Shin ),( Sun-young Lee ),( Jeong Seong Yang ),( Jake Whang ),( Eun-soon Son ),( Tae Won Jang ),( Je Hun Kim ),( Jee Youn Oh ),( Jusang Kim ),( Hyungwoo Kim ),( Yong-soon Cho ) 대한결핵 및 호흡기학회 2021 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.129 No.0
Background Minimum inhibitory concentration (MIC) of antibiotics has been widely conducted for drug susceptibility test (DST) of most bacterial infectious diseases, except tuberculosis (TB). Current phenotypic DST for TB uses one or two critical concentrations to determine the resistance of anti-TB drugs. The MIC test would be necessary not only for therapeutic drug monitoring-based precision medicine but also for detecting borderline resistance of anti-TB drugs. Here, we present the distribution of MIC of anti-TB drugs in cPMTb cohort Mycobacterium tuberculosis (Mtb) isolates. Methods From 2019-2020, 205 Mtb strains were collected in the cPMTb cohort. Broth microdilution method in 96 well plates were used for the MIC test of anti-TB drugs, except for pyrazinamide (PZA). MGIT960 system was used for PZA MIC determination. Results The proportions of drug-resistant (DR) and drug-susceptible (DS) strains were 13.1% and 86.9%, respectively. Most of the DS MIC were distributed below the critical concentration of the drug. Six discordances between phenotypic or genotypic DST and MIC were found in this study. Two cases of disputed mutation L511P on rpoB were included in the discordance cases. Their MIC of rifampicin was 0.5 ug/ml. It is lower than critical concentration but higher than epidemiological cut-off value. Conclusion MIC of cPMTb cohort 205 strains were distributed mostly below the critical concentration of all drugs. There are 6 cases of discordance between MIC and DST (phenotypic or genotypic). The MIC test could be utilized to overcome the limitation of phenotypic DST using critical concentration and fill the gap between genotypic and phenotypic DST.
( Yumi Park ),( Sun-young Lee ),( Eun-soon Son ),( Jeong Seong Yang ),( Jake Whang ),( Tae Won Jang ),( Je Hun Kim ),( Jee Youn Oh ),( Hyn Kuk Kim ),( Hyo-jung Kim ),( Yong-soon Cho ),( Jae-gook Shin 대한결핵 및 호흡기학회 2020 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.128 No.-
Background Minimum inhibitory concentration (MIC) of antibiotics has been widely conducted for drug susceptibility tests (DST) of most bacterial infectious diseases, except tuberculosis (TB). Current phenotypic DST for TB uses one or two critical concentrations to determine the resistance of anti-TB drugs. The MIC test would be necessary not only for therapeutic drug monitoring-based precision medicine but also for detecting borderline resistance of anti-TB drugs. Here, we present the distribution of MIC of 4 first-line drugs and levofloxacin in cPMTb cohort Mycobacterium tuberculosis (Mtb) isolates. Methods From 2019-2020, 108 Mtb strains were collected in the cPMTb cohort. Broth microdilution Method in 96 well plates was used for the MIC test of anti-TB drugs, except for pyrazinamide (PZA). MGIT960 system was used for PZA MIC determination. Results The proportions of drug-resistant (DR) and drug-susceptible (DS) strains were 15.7% and 84.3%, respectively. Most of the DS MIC were distributed below the critical concentration of the drug. Three discordances between phenotypic DST and MIC were found in isoniazid, which is 2.8% in total. We observed 2 cases (1.9%) of low-level resistance (MIC 0.5 ug/ml), determined as DS for rifampicin in phenotypic DST but DR in genotypic DST. Conclusion MIC of cPMTb cohort 108 strains were distributed mostly below the critical concentration of all drugs. There are 6 cases of discordance between MIC and DST (phenotypic or genotypic). The MIC test could be utilized to overcome the limitation of phenotypic DST using critical concentration and fill the gap between genotypic and phenotypic DST.
Lee, Kang-In,Choi, Han-Gyu,Son, Yeo-Jin,Whang, Jake,Kim, Kwangwook,Jeon, Heat Sal,Park, Hye-Soo,Back, Yong Woo,Choi, Seunga,Kim, Seong-Woo,Choi, Chul Hee,Kim, Hwa-Jung Springer-Verlag 2016 Apoptosis Vol.21 No.4
<P>Mycobacterium avium and its sonic extracts induce apoptosis in macrophages. However, little is known about the M. avium components regulating macrophage apoptosis. In this study, using multidimensional fractionation, we identified MAV2052 protein, which induced macrophage apoptosis in M. avium culture filtrates. The recombinant MAV2052 induced macrophage apoptosis in a caspase-dependent manner. The loss of mitochondrial transmembrane potential (Delta I-m), mitochondrial translocation of Bax, and release of cytochrome c from mitochondria were observed in macrophages treated with MAV2052. Further, reactive oxygen species (ROS) production was required for the apoptosis induced by MAV2052. In addition, ROS and mitogen-activated protein kinases were involved in MAV2052-mediated TNF-alpha and IL-6 production. ROS-mediated activation of apoptosis signal-regulating kinase 1 (ASK1)-JNK pathway was a major signaling pathway for MAV2052-induced apoptosis. Moreover, MAV2052 bound to Toll-like receptor (TLR) 4 molecule and MAV2052-induced ROS production, Delta I-m loss, and apoptosis were all significantly reduced in TLR4(-/-) macrophages. Altogether, our results suggest that MAV2052 induces apoptotic cell death through TLR4 dependent ROS production and JNK pathway in murine macrophages.</P>
Byun, Eui-Hong,Kim, Woo Sik,Shin, A-Rum,Kim, Jong-Seok,Whang, Jake,Won, Choul-Jae,Choi, Yohan,Kim, Su-Young,Koh, Won Jung,Kim, Hwa-Jung,Shin, Sung Jae Springer 2012 Journal of molecular medicine Vol.90 No.3
<P>Tuberculosis (TB) caused by Mycobacterium tuberculosis (Mtb) is one of the most deadly infectious diseases, with approximately two million people dying of TB annually. An effective therapeutic method for activating dendritic cells (DCs) and driving Th1 immune responses would improve host defenses and further the development of a TB vaccine. Given the importance of DC maturation in eliciting protective immunity against TB, we investigated whether Rv0315, a newly identified Mtb antigen, can prompt DC maturation. We found that Rv0315 functionally activated DCs by augmenting the expression of the co-stimulatory molecules CD80 and CD86 as well as MHC class I/II molecules. Moreover, it increased DC secretion of the pro-inflammatory cytokines IL-6, IL-1 beta, and TNF-alpha. Unlike LPS, however, Rv0315 induced the secretion of IL-12p70, but not IL-10. In addition, Rv0315-treated DCs accelerated the proliferation of CD4(+) and CD8(+) splenic T cells from Mtb-infected mice, with increased levels of IFN-gamma, in syngeneic and allogeneic mixed lymphocyte reactions, indicating that Rv0315 contributes to Th1 polarization of the immune response. Importantly, both mitogen-activated protein kinases and nuclear factor kappa B signaling mediated the expression of DC surface markers and cytokines. Taken together, our results indicate that Rv0315 is a novel DC maturation-inducing antigen that drives T cell immune responses toward Th1 polarization, suggesting that Rv0315 plays a key role in determining the nature of the immune response to TB.</P>
Comparison of Immune Responses between Smooth and Rough Variant of Mycobacterium Mucogenicum
( Minji Kang ),( Ho Won Kim ),( A-reum Yu ),( Jeong Seong Yang ),( Hyeonji Kim ),( Seung Heon Lee ),( Jong-seok Kim ),( Jake Whang ) 대한결핵 및 호흡기학회 2020 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.128 No.-
Background Mycobacterium mucogenicum (Mmuc), a rapidly growing nontuberculous mycobacterium (NTM), is a rarely causative agent of clinical diseases such as post-traumatic wound infections and catheter-related sepsis in immunocompromised patients. Especially, Mmuc has been identified as a water contaminant in the hospital because it was more resistant to chlorine than other bacteria including M. fortuitum as well as E. coli. Like other NTM species mycobacteria such as M. abscessus, Mmuc has a characteristic to exhibit colony morphologies of rough (R) and variable smooth (S) types. It is known that R type is associated with more severe and persistent infection than S type. However, little is known about the immune responses of Mmuc. Methods To compare the immune responses between S and R type of Mmuc, S types including ATCC 49650 and two clinical strains, and one clinical strain with R type were obtained from the Korean Mycobacteria Resource Center (KMRC) of Korean Institute of Tuberculosis. Therefore, each type of Mmuc were infected to murine bone marrow-derived macrophage (BMDM), then intra-macrophage growth of Mmuc were measured by time dependently. Lastly, we conducted the ELISA of inflammatory cytokine secretion and then confirmed cell death pattern by FACS analysis. Results Macrophages infected with R strain had the higher numbers of intracellular bacteria after 3 days post-infection than S strains. Also, the R strain induced higher levels of cytokine production such as TNF-α, IL-6 and derived more necrotic cell death of BMDM. Conclusions In Conclusion, these Results suggest that R strain has more virulence and induces stronger immune response than S strains likewise other NTM species, MAC and M. abscessus. It is meaningful as the first finding about differential immune responses of Mmuc, also it might provide a fundamental research for further studies on Mmuc pathogenesis.
Prevalence of Mycobacterium Bovis among Human Tuberculosis Specimens in South Korea
( Minji Kang ),( Ho Won Kim ),( A-reum Yu ),( Jeong Seong Yang ),( Hyeonji Kim ),( Jong-seok Kim ),( Jake Whang ),( Seung Heon Lee ) 대한결핵 및 호흡기학회 2020 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.128 No.0
Background Mycobacterium bovis is a zoonotic pathogen that can infect both humans and animals as tuberculosis (TB). Unlikely other Mycobacterium tuberculosis complex (MTBC) members, M. bovis is intrinsically resistant to pyrazinamide as one of the first-line anti-tuberculosis drugs. The World Health Organization reported that an estimated 143,000 new cases of these zoonotic TB occurred globally in 2018. Transmission is mostly through close contacts with infected cattle or consumption of unpasteurized dairy products. Nevertheless, M. bovis infected patients are not distinguished among TB patients, and even while many epidemiologic reports on M. bovis infection have been published in lots of countries, there has never been an attempt to identify M. bovis in Korea. Methods A total of 160 MTBC clinical isolates having pyrazinamide resistance was provided from Korean Mycobacteria Resource Center of Korean Institute of Tuberculosis from 2008 to 2019. We cultured on Middlebrook 7H10 medium and then genomic DNA of mycobacterial isolates was extracted using the boiling Method. Multiplex PCR assay was performed by using the commercial kit. Results We reviewed data on the proportion of M. bovis in human samples published in other 18 countries. Among these, the highest percentage is 30.20 % in Mexico. In contrast, the lowest percentage is 0.13 % in Ethiopia. Our Results have not yet showed any detection as M. bovis among these 107 MTBC isolates in Korea. Conclusions In Conclusion, we reviewed the proportion of M. bovis isolated from human and then we conducted the first differentiation of M. bovis from MTBC isolates in Korea. These Results show that the system of Korean livestock industry is very tightly controlling the infection of M. bovis. However, apart from that, it is also important to continually monitor another route of TB and to complete M. bovis infection statistics through more tests.