Modern History of Hansen's Disease in Korea

Modern history of Hansen's Disease (HD) in Korea begins with nationwide use of the chemotherapeutic agent Diamino Diphenyl Sulphone for the patients in 1955. Definition of the case was different from time to time. Based on World Health Organization (WHO) criteria, Ministry of Health and Welfare (MOHW) reported 4,393 registered patients and same number 4,393 as new cases in 1977. This is the turning point they accepted patient reporting system of WHO, but total number of registered and managed as leprosy patients was 28,029 in 1977, which means the people who needs HD service from government at that time. The number of new cases decreased from 4,393 in 1977, 39 in 1996 to 4 in 2017. Regarding to new cases, it takes 40 years to accomplish from thousands level to below 10. Now we have 166 active cases (registered patients) and reported them as patients to the WHO. Korea Civil Assistance Command invited Dr. RG Cochrane who visited Korea for six weeks to make blue print for eradication of HD in Korea. With his advice and MOHW set HD project and plan for manpower to solve HD problems in 1955. Dr. Joon Lew and his colleagues founded Korean Leprosy Prevention Association in 1947 to combat leprosy, enlighten the public, and solve social problems caused by HD. The Korean Leprosy Prevention Association led by him changed its name to the Korean Leprosy Association in 1956, and grew into the current Korean Hansen Welfare Association. This organization is now playing a leading role in the eradication and management of HD in Korea.

Modern History of Hansen's Disease in Korea

Modern history of Hansen's Disease (HD) in Korea begins with nationwide use of the chemotherapeutic agent Diamino Diphenyl Sulphone for the patients in 1955. Definition of the case was different from time to time. Based on World Health Organization (WHO) criteria, Ministry of Health and Welfare (MOHW) reported 4,393 registered patients and same number 4,393 as new cases in 1977. This is the turning point they accepted patient reporting system of WHO, but total number of registered and managed as leprosy patients was 28,029 in 1977, which means the people who needs HD service from government at that time. The number of new cases decreased from 4,393 in 1977, 39 in 1996 to 4 in 2017. Regarding to new cases, it takes 40 years to accomplish from thousands level to below 10. Now we have 166 active cases (registered patients) and reported them as patients to the WHO. Korea Civil Assistance Command invited Dr. RG Cochrane who visited Korea for six weeks to make blue print for eradication of HD in Korea. With his advice and MOHW set HD project and plan for manpower to solve HD problems in 1955. Dr. Joon Lew and his colleagues founded Korean Leprosy Prevention Association in 1947 to combat leprosy, enlighten the public, and solve social problems caused by HD. The Korean Leprosy Prevention Association led by him changed its name to the Korean Leprosy Association in 1956, and grew into the current Korean Hansen Welfare Association. This organization is now playing a leading role in the eradication and management of HD in Korea.

Expression of Nucleotide-oligomerization Domain (NOD) and Related Genes in Mouse Tissues Infected with Mycobacterium leprae

The nucleotide-oligomerization domain (NOD) is an important molecule involved in host defense against bacterial infection. To study the role of NODs in the host response to Mycobacterium leprae, we measured the mRNA levels of NODs and related genes in infected mouse tissues. The mRNA expression of NOD1, NOD2, caspase-1 and ASC was increased in mouse footpads. Whereas NOD2 expression in macrophages was increased at 2 and 24 h post-infection with M. leprae, there was no expression of NOD1 at these time points. An increase in caspase-1 expression was observed at 2 h and continued at 24 h. However, the expression of ASC was increased only at the early time point. The expression of caspase-1 is regulated by NOD2-dependent pathway in established HEK 293. Our results suggest NOD2, rather than NOD1, may be associated with the host response to M. leprae and that caspase-1 activation is essential for the host response.

The Role of Intracellular Receptor NODs for Cytokine Production by Macrophages Infected with Mycobacterium leprae

The nucleotide-oligomerization domain (NOD) proteins are members of the NOD-like receptor (NLR) family, which are intracellular and cytoplasmic receptors. We analyzed the role of NODs for cytokine production by macrophages infected with intracellular pathogen M. leprae, the causative agent of leprosy. Production of pro-inflammatory cytokines such as IL-1beta and TNF-alpha was inhibited in the presence of cytochalasin D, an agent blocking phagocytosis, suggesting that intracellular signaling was, partially, required for macrophage activation to M. leprae infection. Next, we investigated the role of NOD1 and NOD2 proteins on NF-kappaB activation and cytokine expression. Treatment with M. leprae significantly increased NF-kappaB activation and expression of TNF-alpha and IL-1beta in NOD1- and NOD2-transfected cells. Interestingly, their activation and expression were inhibited by cytochalasin D, suggesting that stimulation of NOD proteins may be associated with the enhancement of cytokine production in host to M. leprae.

Vitamin D Receptor Gene TaqI, BsmI and FokI Polymorphisms in Korean Patients with Tuberculosis

BACKGROUND: The active metabolite (1, 25-dihydroxycholecalciferol) of vitamin D (25-hydroxycholecalciferol) leads to activation of macrophages and deficiency of vitamin D seems to be involved in the risk of tuberculosis. The effects of vitamin D are exerted by interaction with the vitamin D receptor (VDR) and may be influenced by polymorphism in the VDR gene. In this study, variation in the VDR gene was investigated in Korean population with tuberculosis. METHODS: We typed three VDR polymorphisms of restriction endonuclease sites for TaqI, BsmI and FokI in 155 patients with tuberculosis and 105 healthy volunteers. RESULTS: The frequencies of FokI genotypes determined from TB patients were 29.13% for FF, 56.31% for Ff, and 14.56% for ff. We observed 1.4-fold increased prevalence of the Ff genotype in TB patients compared with normal healthy groups (p=0.0857). However, there was no significant association between the genotype groups, TB patient and normal control, for FokI polymorphism. There was also no significant association between VDR gene and tuberculosis in another polymorphism (BsmI and TaqI). CONCLUSION: Three polymorphisms (TaqI, BsmI and FokI) in the VDR gene do not appear to be responsible for host susceptibility to human tuberculosis in Korean population.

Vitamin D Receptor Gene TaqI, BsmI and FokI Polymorphisms in Korean Patients with Tuberculosis

BACKGROUND: The active metabolite (1, 25-dihydroxycholecalciferol) of vitamin D (25-hydroxycholecalciferol) leads to activation of macrophages and deficiency of vitamin D seems to be involved in the risk of tuberculosis. The effects of vitamin D are exerted by interaction with the vitamin D receptor (VDR) and may be influenced by polymorphism in the VDR gene. In this study, variation in the VDR gene was investigated in Korean population with tuberculosis. METHODS: We typed three VDR polymorphisms of restriction endonuclease sites for TaqI, BsmI and FokI in 155 patients with tuberculosis and 105 healthy volunteers. RESULTS: The frequencies of FokI genotypes determined from TB patients were 29.13% for FF, 56.31% for Ff, and 14.56% for ff. We observed 1.4-fold increased prevalence of the Ff genotype in TB patients compared with normal healthy groups (p=0.0857). However, there was no significant association between the genotype groups, TB patient and normal control, for FokI polymorphism. There was also no significant association between VDR gene and tuberculosis in another polymorphism (BsmI and TaqI). CONCLUSION: Three polymorphisms (TaqI, BsmI and FokI) in the VDR gene do not appear to be responsible for host susceptibility to human tuberculosis in Korean population.

Rifampicin Inhibits the LPS-induced Expression of Toll-like Receptor 2 via the Suppression of NF-kappaB DNA-binding Activity in RAW 264.7 Cells

Rifampicin is a macrocyclic antibiotic which is used extensively for treatment against Mycobacterium tuberculosis and other mycobacterial infections. Recently, a number of studies have focused on the immune-regulatory effects of rifampicin. Therefore, we hypothesized that rifampicin may influence the TLR2 expression in LPS-activated RAW 264.7 cells. In this study, we determined that rifampicin suppresses LPS-induced TLR2 mRNA expression. The down-regulation of TLR2 expression coincided with decreased production of TNF-alpha. Since NF-kappaB is a major transcription factor that regulates genes for TLR2 and TNF-alpha, we examined the effect of rifampicin on the LPS-induced NF-kappaB activation. Rifampicin inhibited NF-kappaB DNA-binding activity in LPS-activated RAW 264.7 cells, while it did not affect IKKalpha/beta activity. However, rifampicin slightly inhibited the nuclear translocation of NF-kappaB p65. In addition, rifampicin increased physical interaction between pregnane X receptor, a receptor for rifampicin, and NF-kappaB p65, suggesting pregnane X receptor interferes with NF-kappaB binding to DNA. Taken together, our results demonstrate that rifampicin inhibits LPS-induced TLR2 expression, at least in part, via the suppression of NF-kappaB DNA-binding activity in RAW 264.7 cells. Thus, the present results suggest that the rifampicin-mediated inhibition of TLR2 via the suppression of NF-kappaB DNA-binding activity may be a novel mechanism of the immune-suppressive effects of rifampicin.

Comparison of rpoT gene of M. leprae strain from korean and foreign leprosy patients / 나학회지

The variance of tandem repeats in the rpoT gene of Mycobacterium leprae was recently demonstrated. The objects of this study was to examine the proportion and distributions of the genotypes of M. leprae in Korea and to compared it with genotypes of M. leprae form foreign leprosy patients using difference of the tandem repeats. Among 101 cases, 72 isolated from Korea and 4 cases from Japan (except Okinawa) demonstrated four copies of the 6 bp tandem repeats in the rpoT gene, and three copies were found in isolates from two korean, 2 cases of Okinawa in Japan, and those from Southeast Asian countries, Peru and Paraguay. These results reveal the genetic diversity of M. leprae and the related genotype-specific distribution in the world. In this study, a more detailed explanation can be also possible regarding the transmission route of M. leprae.

Mutations in folP1, rpoB, and gyrB associated with multiple drug resistance in Mycobacterium leprae isolates from South Korea / 나학회지

The emergence of multiple drug resistant Mycobacterium leprae has emphasized the need for early decision of effective antileprosy drug in the treatment for leprosy patients. Mutations in the genes associated with multiple drug resistance in Mycobacterium leprae isolates from 17 South Korean patients, who were already confirmed to have mutations in folP1 gene, were investigated using a PCR - single strand conformation polymorphism (SSCP) - DNA sequencing assay. Two strains, which has double mutations, were found in the two unrelated patients : one missense mutation in folP1 (Arg 55 for Pro) and in rpoB (Gly 522 for Ser), and in folP1 (Ala 53 for Thr) and in gyrB (Asn 426 for Asp), respectively. The patients were treated with the long monotheraphy of dapsone or with the inappropriate regimen of antileprosy drugs. These results emphasize the importance of multi-drug theraphy in order to prevent mult-idrug resistance and assist in the choice of the appropriate regimens for treating leprosy.

The association study on infection of Mycobacterium leprae and RIPK2 / 나학회지

Receptor-interacting serine/threonine kinase 2(RIPK2) is an adaptor molecule involved in the signal pathway of TLRs. However, there is no report on association between RIPK2 expression and infectious disease including mycobacterial disease in which TLRs play main role on interaction of infection. We evaluated relationship between Mycobacterium leprae and RIPK 2 by real-time RT-PCR. This study revealed that RIPK2 expression was down-regulated in the footpads and skin but was up-regulated in the liver, lymph node, and spleen of Mycobacterium leprae-infected nu/nu mice compared with those of non-infected nu/nu mice. It was observed that the IL-12p40, IFN-gamma, and IL-18 involved in the susceptibility of Mycobacterium leprae were down-regulated in the skin and footpad but up-regulated in the liver. These results suggest that regulation of RIPK2 expression is tissue-specific and is associated with M. leprae infection.

Detection of point mutation at C-terminal region of phagosomal coat protein (TACO) in patients with leprosy / 나학회지

Mycobacteria, which are highly successful pathogen, resist delivary to lysosomes and instead survive within a specialized vacuole, the mycobacterial phagosome. The bacteria survive intracellularly because they are able to actively recruit and retain TACO ( tryptophane aspartate-containing coat protein ) at the mycobacterial phagosome, where it prevents lysosomal delivary in a cholesterol-dependent manner. In this study, we investigated the difference of TACO expression is whether related to mutant in coro1a gene in patients with leprosy and normal volunteer. First, we screened for detection of a mutant in the leucine zipper motif within the exon 11, and then in the exon 9 to 10, and finally in the coiled-coil region. Interestingly, single base substitutions ( point mutation ) presents at assembly site of U1 snRNP, around of 5' splice site in the intron 9, there are a C to T and G to A transition are at 9 bp and 14 bp downstream of 5' splice site, respectively, and both of it. Among the 3 types of polymorphism, frequency of a G to A transition is markedly increased in patients of lepromatous type, which are new cases or relapsed. Both a C to T and G to A transitions are found in 1 case of tuberculoid type and 2 cases in lepromatoue type, but not found in control group. The silent mutation in leucine zipper motif within the exon 11 is located at codon at 454 ( CTG-->CTA), which is 1st leucine from C-terminal among four leucine zipper. In coiled-coil region, no mutation is found in genomic DNA of patients with leprosy. Further, we will do functional study about the identified point mutation and will screen any possible mutation in the region of promotor and WD repeat.

Effects of Dexamethasone on expressions of IFN-gamma and IL-4 by PBMCs in response to IL-12 / 나학회지

Dexamethasone is a widely used anti-inflammatory agent for a broad spectrum of diseases. The effectiveness of this agent is thought to be due to the capacity to modulate cytokine production in inflammatory cells. We examined the effects of dexamethasone on expressions of interferon gamma (IFN-gamma) and interleukin 4 (IL-4) by human peripheral blood mononuclear cells (PBMCs) in response to interleukin 12 (IL-12). Dexamethasone (10-5 M) inhibited IFN-gamma secretion, through direct suppression of IFN-gamma, IL-12 receptor (IL-12R) -beta1, and -beta2 expressions. Conversely dexamethasone increased IL-4 secretion as well as IL-4 expressions by PBMCs in response to IL-12. In addition, dexamethasone increased expression of suppressor of cytokine signalling (SOCS)-1, which inhibits JAK-STAT pathway of IL-12R signalling. The result of our study suggested that dexamethasone directly inhibited IFN-gamma expression, through suppression of IL-12 signalling and indirectly increases IL-4 expression, through suppression of IFN-gamma expression.

The Development Tasks of Medical School Accreditation in Korea / 한국의학교육

The Accreditation Board for Medical Education in Korea, ABMEK, is nongovernmental appraisal organization that was established at July 2, 1998. The organization is contributing to the improvement of medical education by progressing the first cycle accreditation successfully. But, the organization has various problems and subjects related to the accreditation system. The authors examined the related literature focusing on the current status and problems of accreditation system. The result of this research was as follows. First, the ABMEK needs to propel legal personality of organization and should install independent executive office. Second, the ABMEK should establish the alteration procedure of accreditation standards and develop the accreditation standards of the second cycle that take into account international flowing of medical education. Third, the ABMEK must decide forms and scope to investigate medical college present situation. Finally, to propel development tasks effectively, it needs to get the recognition of Ministry of Education and Human Development.

Differentiation of Osteoblast Progenitor Cells from Human Umbilical Cord Blood

BACKGROUND: Human umbilical cord bloods, which could be taken during the delivery are utilized as a source of hematopoietic stem cells. Also in cord blood, there are several kinds of stem cells such as endothelial and mesenchymal stem cells. METHODS: We isolated the mesenchymal stem cells from human umbilical cord bloods and confirmed the differentiation of these cells into osteoblast progenitor cells. The mesenchymal stem cells derived from umbilical cord blood have the ability to differentiate into specific tissue cells, which is one of characteristics of stem cells. These cells were originated from the multipolar shaped cells out of adherent cells of the umbilical cord blood mononuclear cell culture. RESULTS: The mesenchymal stem cells expressed cell surface antigen CD13, CD90, CD102, CD105, alpha-smooth muscle actin and cytoplasmic antigen vimentine. Having cultrued these cells in bone formation media, we observed the formation of extracellular matrix and the expression of alkaline phosphatase and of mRNA of cbfa-1, ostoecalcin and type I collagen. CONCLUSION: From these results we concluded that the cells isolated from the umbilical cord blood were mesenchymal stem cells, which we could differentiate into osteoblast when cultured in bone formation media. In short, it is suggested that these cells could be used as a new source of stem cells, which has the probability to alternate the embryonic stem cells.

Differentiation of Osteoblast Progenitor Cells from Human Umbilical Cord Blood

BACKGROUND: Human umbilical cord bloods, which could be taken during the delivery are utilized as a source of hematopoietic stem cells. Also in cord blood, there are several kinds of stem cells such as endothelial and mesenchymal stem cells. METHODS: We isolated the mesenchymal stem cells from human umbilical cord bloods and confirmed the differentiation of these cells into osteoblast progenitor cells. The mesenchymal stem cells derived from umbilical cord blood have the ability to differentiate into specific tissue cells, which is one of characteristics of stem cells. These cells were originated from the multipolar shaped cells out of adherent cells of the umbilical cord blood mononuclear cell culture. RESULTS: The mesenchymal stem cells expressed cell surface antigen CD13, CD90, CD102, CD105, alpha-smooth muscle actin and cytoplasmic antigen vimentine. Having cultrued these cells in bone formation media, we observed the formation of extracellular matrix and the expression of alkaline phosphatase and of mRNA of cbfa-1, ostoecalcin and type I collagen. CONCLUSION: From these results we concluded that the cells isolated from the umbilical cord blood were mesenchymal stem cells, which we could differentiate into osteoblast when cultured in bone formation media. In short, it is suggested that these cells could be used as a new source of stem cells, which has the probability to alternate the embryonic stem cells.

Changes of Leprosy Research in the Post Genomic Era / 나학회지

Leprosy has turned out to be a minor health problem in Korea due to implementation of multidrug therapy, intensive case detection and mobile works for the villages for last 20 years. Even we have 20 new cases a year and national registry of 18,000, elimination of leprosy in Korea will require more decades in comparison with Japan, which seems to be an advanced country in leprosy work but still has new cases not more than 10 a year. As genome of M. leprae has unravelled to public in 2001, molecular biologic and genetic research work in leprosy area will progress rapidly near future. Range of leprosy research depends on the state of personal and grant resources in the nation. As Korea has only limited scientist, clinicians and financial support for research work to solve problems in leprosy. To maintain research capabilities and man powers in leprosy, it is reasonable to adjust our research works for priorities of the problems in leprosy work in Korea. Diagnosis, follow up and detection of drug resistance in new cases using molecular epidemiologic and genetic tools are one of high priorities in Korea. Management of neuritis and prevention of deformities on eyes, hands and foot of post-period of multidrug therapy appear to be an important area to improve the quality of life of the treated patients. Collaboration with researchers in tuberculosis, adoption of methods from them can expand research capacity and keep leprosy research alive in Korea.

Optimization for cell lysate preparation of M. leprae from infected nude mouse / 나학회지

The method of cell lysate preparation of M. leprae is an important technique in the study of leprosy. This report describes the optimization of method for cell lysate preparation of M. leprae obtained from infected nude mouse. After M. leprae isolated from nude mouse foot-pad was disrupted by sonication, it was centrifuged and then whole lysate was prepared. With this method it was possible to isolate 0.3 mg whole cell lysate using 20 mg of M. leprae. By SDS-PAGE and Coomassie blue staining, the number of protein in M. leprae is less than that of other bacteria, for example, E. coli and M. smegmatis. It is likely that this is due to the small genome size. This work will contribute to the analysis of new protein antigen of M. leprae and the basic study for the development of vaccine in leprosy.

The Change of Bacillary Index during WHO Standard Multi-drug Therapy in Multi-bacillary Leprosy: Among the patients in The National Sorokdo Hospital / 대한피부과학회지

BACKGROUND: There were no effective methods to monitor the treatment of leprosy. Anti-PGL-1 ELISA test and polymerase chain reaction(PCR) were used for monitoring the treatment, however many restrictions have been found to apply to those methods in the field. Bacillary index(BI) is a popular and not an expensive method. For this reason, the exact data to assess the efficacy of the standard multi-drug therapy(MDT) with BI is required for evaluation of the treatment in Korea. OBJECTIVE: The purpose of this study was to clarify the change of bacillary index during MDT in multi-bacillary leprosy(MB) cases. METHODS: A total of 72 patients in the National Sorokdo Hospital were included in this study. With the retrospective method, BI, type of leprosy, relapse, age, and duration of treatment were evaluated. RESULTS: The results were as follows; 1.In this study, the mean duration of BI changing from 6 to 5 was 5.7 months, from 5 to 4 was 6.6 months, from 4 to 3 was 13.1 months, from 3 to 2 was 13.8 months, from 2 to 1 was 12.4 months, from 1 to 0 was 29.4 months. 2.The time of initial change of BI after MDT were as follows: within 3 months(26.4%), 4-6 months(29.2%), 7-12 months(26.4%), 13-18 months(8.3%), 19-24 months(4.2%), 25-48 months(5.5%). The mean duration was 8.9 months. 3.The mean cumulative time of BI to 0 was 68.4 months CONCLUSION: These results suggest that after MDT, in most of MB, BI decreased within one year and it took 68.4 months for BI to be 0. At first, BI decreased relatively fast, however a longer period was required to become a state of BI 0.

The effect of TLR2 mutation in IL-12 production according to type of leprosy / 나학회지

Recently, many reports suggest that the activation of Toll-like receptor 2 (TLR2) by microbial lipoproteins may initiate innate defense mechanism against infectious pathogens. Especially, TLR2 is critical in the immune response to mycobacterial infections and the mutations in the TLR2 have been shown to confer the susceptibility to severe infection with mycobacteria. A previous study reported that there is a mutation of TLR2 (TLR2-R677W) in lepromatous leprosy. In this study, thus, we performed the functional study on TLR2 by measurement of IL-12 production in serum and monocytes from leprosy patients with TLR2 mutation (R677W) to verify the relation between TLR2 and the susceptibility to leprosy. Tuberculoid leprosy patients showed higher serum levels of IL-12p70 compared with those of lepromatous leprosy. In contrast with the lepromatous leprosy patient with TLR2 mutation, especially, the group with wild-type TLR2 showed 2-fold increase levels of IL-12p70. Functional studies demonstrated that monocytes from patient with the TLR2 mutation, in comparison to the wild-type TLR2, is significantly less responsive to Mycobacterium leprae lysate. Our results reveal that TLR2 has a important role in the IL-12 production from monocytes and the susceptibility of lepromatous leprosy.

The application of TD (Touch-Down) PCR in Diagnosis of leprosy and Detection of MDT-resistant M. leprae / 나학회지

There are several methods for diagnosis of leprosy, including AFB stain, the measurement of PGL-1 (phenolic glycolipid - 1) antigen titer, and DNA-PCR. In this study, we have used the DNA-PCR amplifying the RLEP repetitive sequence. Our result showed that the RLEP primer offered the more sensitive detection and identification of M. leprae DNA in clinical specimens, compared with the other primer, for example, 18-kDa antigen gene. To screen the resistant M. leprae strain of MDT (Multi-Drug Therapy), we have used the TD (Touch-Down) PCR. We arranged and amplified sequences of the genes, folP, rpoB, gyr, 23S rRNA, in M. leprae involved in MDT-resistance, and could obtain the PCR product each gene, simultaneously. This method, based on annealing temperature, was useful to the detection for diagnosis and the screen of MDT-resistant strain of M. leprae, rapidly. Thus, we suggest that the RLEP primer and TD-PCR method are effective in assessing the diagnosis of leprosy and the identification of drug-resistant M. leprae.