Utility of immunoglobulin isotypes against LID-1 and NDO-LID for, particularly IgG1, confirming the diagnosis of multibacillary leprosy.

BACKGROUND Leprosy remains a health problem in many countries, with difficulties in diagnosis resulting in delayed treatment and more severe disabilities. Antibodies against several Mycobacterium leprae antigens have, however, shown value as diagnostic and/or prognostic markers. OBJECTIVES The objective of this study was to evaluate serum immunoglobulin (Ig) IgM and IgG subclass reactivity against three M. leprae specific antigens: NDO-HSA, a conjugate formed by natural octyl disaccharide bound to human serum albumin; LID-1, the fusion protein product of the ml0405 and ml2331 genes; and NDO-LID, a combination of LID-1 and NDO. METHODS Sera from healthy controls, paucibacillary (PB) and multibacillary (MB) leprosy patients, and their respective household contacts, were evaluated for the presence of antigen-specific IgM, IgG, and IgG subclass antibodies by enzyme-linked immunosorbent assay (ELISA). The sensitivity and specificity of each ELISA were evaluated by receiver operating characteristic (ROC) curve analysis. FINDINGS Our data confirm that serum IgM antibodies against NDO-HSA and IgG antibodies against LID-1, as well as IgG/M antibodies against NDO-LID, are markedly increased in MB patients. For the first time, our data reveal a selective increase in IgG1 and IgG3 antibodies against LID-1 and NDO-LID in MB patients, demonstrating that these antibody isotypes are suitable for differentiation between MB and PB patients. ROC curve analysis indicates an improved capacity for diagnosing MB leprosy patients using the detection of IgG antibodies, particularly the IgG1 isotype, specific to LID-1 and NDO-LID over the performance levels attained with NDO-HSA. CONCLUSIONS Our findings indicate that serological tests based on the detection of antigen-specific IgG1 antibodies are a useful tool to differentiate MB from PB patients, and indicate the enhanced performance of the LID-1 and NDO-LID antigens in the serodiagnosis of leprosy.

Testes sorológicos anti-NDO-HSA, anti-LID-1 e anti-NDO-LID em contatos domiciliares de área não endêmica de hanseníase / Anti-NDO-HSA, anti-LID-1 and anti-NDO-LID serological tests in household contacts of a non-endemic leprosy area

Introdução: Diversos fatores podem interferir no desenvolvimento da hanseníase, entre eles fatores genéticos, convívio com o caso de hanseníase e classificação operacional do caso. Testes sorológicos que avaliam a reatividade de anticorpos IgM e IgG frente a antígenos específicos para o Mycobacterium leprae (M. leprae) podem atuar como auxiliares na vigilância dos contatos e/ou população de risco. Objetivo: Analisar o comportamento dos testes sorológicos anti-PGL-1 sintético (NDO-HSA), anti-LID-1 e anti-NDO-LID em área não endêmica de hanseníase e sua relação com características do caso de hanseníase. Material e métodos: Trata-se de um estudo transversal, do tipo analítico, realizado com 35 contatos domiciliares (CD) dos casos de hanseníase. A coleta de dados ocorreu no período de agosto/2016 a fevereiro/2017 por meio de visitas domiciliares. A reatividade de anticorpos IgM e IgG frente aos antígenos Natural disaccharide linked to human serum albumin via octyl (NDO-HSA), Leprosy IDRI diagnostic 1 (LID-1) e Natural disaccharideoctyl - Leprosy IDRI Diagnostic 1(NDO-LID) foi avaliada por ensaio imunoenzimático (ELISA). Os dados foram exportados e analisados no software StatisticalPackage for the Social Sciences (SPSS®) 24 for Windows. Resultados: Foi observada maior proporção de positividade aos testes em CD de casos multibacilares (MB), que residiam com o caso de hanseníase na época do diagnóstico e que tinham parentesco consanguíneo com o caso. Esses casos de hanseníase MB também apresentaram soropositividade frente aos antígenos testados. O valor do índice ELISA foi maior no grupo de CD de casos MB. Houve concordância moderada e significativa (K= 0,53; p< 0,0001) entre os testes anti-NDO-HSA e anti-NDO-LID, mas não foi detectada diferença entre os testes anti-NDO-HSA e anti-LID-1 (K= -0,05; p= 0,678). A correlação foi positiva entre os três antígenos, porém, entre LID-1 e NDO-HSA, não houve significância estatística (p<0,186). Conclusão: Os resultados sugerem que testes sorológicos em conjunto com as características avaliadas nos contatos domiciliares em área não endêmica de hanseníase,podem atuar como auxiliares na detecção de indivíduos infectados pelo M. leprae, contribuindo para vigilância dos contatos domiciliares

Introduction: Several factors may interfere in the development of leprosy, including genetic factors, conviviality with leprosy patients and operational classification of the case. Serological tests performed to evaluate the reactivity of IgM and IgG antibodies response against Mycobacterium leprae (M. leprae) specific antigens may be used as auxiliary tools for transmission surveillance and/or population at risk. Objective: To analyze the performance of anti-PGL-1 (NDO-HSA), anti-LID-1 and anti-NDO-LID serological tests in non-endemic area of leprosy and the relationship with characteristics of the leprosy case. Material and methods: This is a cross-sectional analytical study of 35 household contacts (HC) of leprosy cases. Data collection was carried out from August 2016 to February 2017 with home visits. The reactivity of IgM and IgG antibodies to Natural disaccharide linked to human serum albumin via octyl (NDO-HSA), Leprosy IDRI diagnostic 1 (LID-1) and Natural disaccharide octyl - Leprosy IDRI Diagnostic 1 (NDO-LID) was evaluated through enzyme-linked immunosorbent assay (ELISA). Data were exported and analyzed using Statistical Package for Social Sciences (SPSS®) 24 for Windows. Results: A higher proportion of positivity was observed in the HC tests of multibacillary (MB) leprosy cases who lived in the same dwelling with a leprosy case at the time of diagnosis and had a degree of kinship with the case. These multibacillary leprosy cases also showed seropositivity to the antigens tests. ELISA test index value was higher in the HC group of MB leprosy cases. There was moderate agreement (K = 0.53, p <0.0001) between anti-NDO-HSA and anti-NDO-LID tests, but no difference was found between anti-NDO-HSA and anti-LID -1 (K = -0.05, p = 0.678). Three antigens were positively correlated, but there was no statistical significance (p <0.186) between LID-1 and NDO-HSA. Conclusion: The results suggest that serological tests in combination with the characteristics assessed during household contacts in a non-endemic area may represent efficient auxiliary tools for the detection of M. leprae-infected individuals, providing a contribution to the surveillance of household contacts

Utility of immunoglobulin isotypes against LID-1 and NDO-LID for, particularly IgG1, confirming the diagnosis of multibacillary leprosy

BACKGROUND Leprosy remains a health problem in many countries, with difficulties in diagnosis resulting in delayed treatment and more severe disabilities. Antibodies against several Mycobacterium leprae antigens have, however, shown value as diagnostic and/or prognostic markers. OBJECTIVES The objective of this study was to evaluate serum immunoglobulin (Ig) IgM and IgG subclass reactivity against three M. leprae specific antigens: NDO-HSA, a conjugate formed by natural octyl disaccharide bound to human serum albumin; LID-1, the fusion protein product of the ml0405 and ml2331 genes; and NDO-LID, a combination of LID-1 and NDO. METHODS Sera from healthy controls, paucibacillary (PB) and multibacillary (MB) leprosy patients, and their respective household contacts, were evaluated for the presence of antigen-specific IgM, IgG, and IgG subclass antibodies by enzyme-linked immunosorbent assay (ELISA). The sensitivity and specificity of each ELISA were evaluated by receiver operating characteristic (ROC) curve analysis. FINDINGS Our data confirm that serum IgM antibodies against NDO-HSA and IgG antibodies against LID-1, as well as IgG/M antibodies against NDO-LID, are markedly increased in MB patients. For the first time, our data reveal a selective increase in IgG1 and IgG3 antibodies against LID-1 and NDO-LID in MB patients, demonstrating that these antibody isotypes are suitable for differentiation between MB and PB patients. ROC curve analysis indicates an improved capacity for diagnosing MB leprosy patients using the detection of IgG antibodies, particularly the IgG1 isotype, specific to LID-1 and NDO-LID over the performance levels attained with NDO-HSA. CONCLUSIONS Our findings indicate that serological tests based on the detection of antigen-specific IgG1 antibodies are a useful tool to differentiate MB from PB patients, and indicate the enhanced performance of the LID-1 and NDO-LID antigens in the serodiagnosis of leprosy.

Detection of IgG1 antibodies against Mycobacterium tuberculosis DosR and Rpf antigens in tuberculosis patients before and after chemotherapy.

Diagnosis of tuberculosis (TB) remains challenging. Serum IgG1 antibodies against Mycobacterium tuberculosis active growth phase antigens (ESAT-6/CFP-10, Rv0717 and Rv3353), DosR regulon-encoded proteins (Rv1733, Rv1737, Rv2628 and Rv2029), and resuscitation-promoting factors (Rv0867 and Rv2389) were evaluated in TB patients using ELISA. Active TB patients showed elevated levels of IgG1 antibodies against ESAT-6/CFP-10, Rv0717, Rv3353, Rv1733, Rv2628, Rv2029 and Rv0867 in comparison to healthy controls (p < 0.001). These levels remained high after the initiation of treatment, while responses to Rv0717 and Rv1733 peaked early during treatment. IgG1 responses to ESAT-6/CFP-10, Rv3353, Rv2628, Rv2029 and Rv0867 declined to control levels after the completion of 6 months chemotherapy. ROC analysis confirmed the good diagnostic performance of Rv0717, Rv1733, Rv3353, Rv2628, Rv2029 and Rv0867antigens. These data suggest that detecting IgG1 antibodies against M. tuberculosis antigens, including DosR and Rpf proteins, may represent an additional tool in the diagnosis of tuberculosis.

Challenging Mycobacterium tuberculosi s dormancy mechanisms and their immunodiagnostic potential

ABSTRACT Mycobacterium tuberculosis is the etiologic agent of tuberculosis, one of the world's greatest cause of morbidity and mortality due to infectious disease. Many evolutionary mechanisms have contributed to its high level of adaptation as a host pathogen. Prior to become dormant, a group of about 50 genes related to metabolic changes are transcribed by the DosR regulon, one of the most complex and important systems of host-pathogen interaction. This genetic mechanism allows the mycobacteria to persist during long time periods, establishing the so-called latent infection. Even in the presence of a competent immune response, the host cannot eliminate the pathogen, only managing to keep it surrounded by an unfavorable microenvironment for its growth. However, conditions such as immunosuppression may reestablish optimal conditions for bacterial growth, culminating in the onset of active disease. The interactions between the pathogen and its host are still not completely elucidated. Nonetheless, many studies are being carried out in order to clarify this complex relationship, thus creating new possibilities for patient approach and laboratory screening.

Challenging Mycobacterium tuberculosis dormancy mechanisms and their immunodiagnostic potential.

Mycobacterium tuberculosis is the etiologic agent of tuberculosis, one of the world's greatest cause of morbidity and mortality due to infectious disease. Many evolutionary mechanisms have contributed to its high level of adaptation as a host pathogen. Prior to become dormant, a group of about 50 genes related to metabolic changes are transcribed by the DosR regulon, one of the most complex and important systems of host-pathogen interaction. This genetic mechanism allows the mycobacteria to persist during long time periods, establishing the so-called latent infection. Even in the presence of a competent immune response, the host cannot eliminate the pathogen, only managing to keep it surrounded by an unfavorable microenvironment for its growth. However, conditions such as immunosuppression may reestablish optimal conditions for bacterial growth, culminating in the onset of active disease. The interactions between the pathogen and its host are still not completely elucidated. Nonetheless, many studies are being carried out in order to clarify this complex relationship, thus creating new possibilities for patient approach and laboratory screening.

Antigen-specific assessment of the immunological status of various groups in a leprosy endemic region.

BACKGROUND: Serological tests can be important tools to assist in the diagnosis of leprosy and can contribute to an earlier diagnosis. The aim of this study was to evaluate the antibody responses against phenolic glycolipid-1 (PGL-1), natural disaccharide linked to human serum albumin via an octyl (NDO-HSA), Leprosy IDRI Diagnostic-1 (LID-1) and natural disaccharide octyl--Leprosy IDRI Diagnostic-1 (NDO-LID) in leprosy patients, household contacts of patients and the general population. METHODS: Enzyme-linked immunosorbent assays were used to analyze the antigen-specific antibody responses of 94 leprosy cases, 104 household contacts of cases and 2.494 individuals from the general population. RESULTS: A positive correlation was observed for the antibody responses to all antigens studied. A higher proportion of seropositivity for all antigens, along with stronger magnitude of response, was observed in multibacillary (MB) leprosy patients and household contacts of MB leprosy patients compared with the levels observed in paucibacillary (PB) leprosy patients and household contacts of PB leprosy patients. A substantial and significant positive correlation was found between seropositivity and the bacterial index for the leprosy patients. Anti-PGL-1 tests were more frequently positive than anti-NDO-HSA tests among patients with all clinical forms of leprosy and among the group of household contacts. The LID-1 and NDO-LID antigens showed a greater capacity to identify household contacts and individuals from the general population infected with M. leprae. CONCLUSIONS: Tests that measure the antibody responses against LID-1, NDO-LID, NDO-HSA and PGL-1 were effective tools for the detection of patients with MB leprosy. Our data indicate that the anti-LID-1 and anti-NDO-LID responses were more effective than an anti-NDO-HSA response for the identification of individuals with subclinical infection.

Biomphalaria spp. (Preston, 1910) snails in the municipality of Juiz de Fora, Zona da Mata Mineira mesoregion, ate of Minas Gerais, Brazil

This study focuses on the geographic distribution of the snail of the genus Biomphalaria and evaluates its infectivity by Schistosoma mansoni in 5264 specimens collected in the municipality of Juiz de Fora, Minas Gerais, Brazil. Of the 31 locations studied, 6 were reservoirs, 11 rudimentary holding ponds, 7 irrigation ditches, 5 lakes, 1 ornamental pond, and 1 waterfall. Intermediate hosts were found only in the rudimentary ponds and ditches, which were 100 percent positive. Using morphological and molecular analysis techniques, B. tenagophila, B. peregrina, and B. straminea were identified. This is the first report of B. stramínea in the municipality, and evaluation of its infective potential revealed susceptibility of 25.4 percent. Although we did not find specimens of Biomphalaria infected by S. mansoni, the data obtained indicate the presence of intermediate hosts, especially in the irrigation ditches in Juiz de Fora, and their proximity to contaminated areas.

Biomphalaria spp. (Preston,1910) snails in the municipality of Juiz de Fora, Zona da Mata Mineira mesoregion, state of Minas Gerais, Brazil.

This study focuses on the geographic distribution of the snail of the genus Biomphalaria and evaluates its infectivity by Schistosoma mansoni in 5264 specimens collected in the municipality of Juiz de Fora, Minas Gerais, Brazil. Of the 31 locations studied, 6 were reservoirs, 11 rudimentary holding ponds, 7 irrigation ditches, 5 lakes, 1 ornamental pond, and 1 waterfall. Intermediate hosts were found only in the rudimentary ponds and ditches, which were 100% positive. Using morphological and molecular analysis techniques, B. tenagophila, B. peregrina, and B. straminea were identified. This is the first report of B. stramínea in the municipality, and evaluation of its infective potential revealed susceptibility of 25.4%. Although we did not find specimens of Biomphalaria infected by S. mansoni, the data obtained indicate the presence of intermediate hosts, especially in the irrigation ditches in Juiz de Fora, and their proximity to contaminated areas.