Expression of NLRP3 inflammasome in leprosy indicates immune evasion of Mycobacterium leprae

BACKGROUND Leprosy is an infectious-contagious disease caused by Mycobacterium leprae that remain endemic in 105 countries. This neglected disease has a wide range of clinical and histopathological manifestations that are related to the host inflammatory and immune responses. More recently, the inflammasome has assumed a relevant role in the inflammatory response against microbiological agents. However, the involvement of inflammasome in leprosy remains poorly understood. OBJECTIVES The aim is to associate biomarkers of inflammasome with the different immunopathological forms of leprosy. METHODS We performed an observational, cross-sectional, and comparative study of the immunophenotypic expression of inflammasome-associated proteins in immunopathological forms of leprosy of 99 skin lesion samples by immunohistochemistry. The intensity and percentage of NLRP3, Caspase-1, Caspases-4/5, interleukin-1β and interleukin-18 immunoreactivities in the inflammatory infiltrate of skin biopsies were evaluated. FINDINGS Strong expression of NLRP3 and inflammatory Caspases-4/5 were observed in lepromatous leprosy (lepromatous pole). In addition, were observed low expression of caspase-1, interleukin-1β, and interleukin-18 in tuberculoid and lepromatous leprosy. The interpolar or borderline form showed immunophenotype predominantly similar to the lepromatous pole. MAIN CONCLUSIONS Our results demonstrate that the NLRP3 inflammasome is inactive in leprosy, suggesting immune evasion of M. leprae.

Leprosy: prevalence and factors associated with seropositivity for anti-NDO-LID antibodies in children under 15 years of age

Abstract: Background: A high prevalence of leprosy among children under 15 years of age indicates the need to implement actions to prevent new cases of the disease. Serological tests have been developed with the aim of helping to control the disease by indicating, through seropositivity, the presence of infection. Objective: To analyze the prevalence and factors associated with seropositivity rate for anti-NDO-LID antibodies in children under 15 years of age, contacts of leprosy patients. Method: We performed a cross-sectional study with 210 children under 15 years old of age. Of them, 50 were household contacts and 160 were neighborhood contacts living in the municipality of Cuiabá, state of Mato Grosso, in 2016. The data were obtained from interviews and the NDO-LID rapid test during home visits from February to July 2016. For the analysis, we used Poisson regression and prevalence ratio. Results: Seropositivity in contacts was 6.2%. Variables associated with seropositive tests included sex (PR = 1.05; 95% CI: 1.01 - 1.08), race/skin color (PR = 0.95; 95% CI: 0.90 - 0.99), residence area (PR = 1.05; 95% CI: 1.01 - 1.09), and number of people per household (PR = 1.06; 95% CI: 1.02 - 1.08). Study Limitations: The small sample size, besides leading to wide confidence intervals, may have been a limitation for the identification of associated factors. Conclusions: The prevalence of seropositivity was high. Variables associated with NDO-LID seropositivity included female sex, not to be brown skinned, live in urban areas, and live with five or more people.

Clinico-immunological spectrum of American tegumentary leishmaniasis and leprosy coinfection: A case series in Southeastern Brazil

Abstract INTRODUCTION: American tegumentary leishmaniasis (ATL) and leprosy share common areas of prevalence, but reports of coinfection are scarce. METHODS: We report a series of 9 ATL-leprosy cases and discuss the association. An integrative diagram to analyze the clinico-immunological features of coinfection with both diseases. RESULTS: Nine patients with leishmaniasis (5 cutaneous, 3 mucocutaneous, 1 disseminated case) exhibited concurrent infection with distinct clinical forms of leprosy. Our diagram-based analysis evidenced a divergent clinico-immunological spectrum for each disease in 8 out of 9 cases. CONCLUSIONS: The spectrum of ATL-leprosy comorbidity suggests that the host has a specific immune response against each pathogen.

Polymorphisms in genes TLR1, 2 and 4 are associated with differential cytokine and chemokine serum production in patients with leprosy

BACKGROUND Leprosy or hansen’s disease is a spectral disease whose clinical forms mostly depends on host’s immune and genetic factors. Different Toll-like receptors (TLR) variants have been described associated with leprosy, but with some lack of replication across different populations. OBJECTIVES To evaluate the role of polymorphisms in genes TLR1, TLR2 and TLR4 and susceptibility to leprosy in a genetic case control study; to verify the association between genotypes of these markers and the immunological profile in the serum of patients with leprosy. METHODS Pre-designed TaqMan® assays were used to genotype markers at TLR1 (rs4833095, rs5743551), TLR2 (rs7656411, rs3804099) and TLR4 (rs1927914, rs1927911). A panel of cytokines and chemokines was accessed by enzime-linked immunosorbent assay (ELISA) test in the serum of a subgroup of patients with and without leprosy reactions. FINDINGS Our results show an association between the T allele of rs3804099 at the TLR2 gene and increased risk for leprosy per se [Odds ratio (OR) = 1.296, p = 0,022]. In addition, evaluating the association between different genotypes of the TLR1, 2 and 4 markers and cytokine/chemokine serological levels, IL-17 appears as an immunological marker regulated by the polymorphism of the three TLR genes evaluated, whereas different TLR1 genotypes were associated with differential production of IL-12p40 and MCP-1(CCL2). Furthermore, other relevant serum markers such as CXCL-10 and IL-6 seemed to be regulated by TLR2 variants and IL-1β was related to TLR4 genotypes. MAIN CONCLUSIONS All together our data points that the tested TLR markers may have a regulatory role in the immunity against Mycobacterium leprae, by driving the host’s production of key cytokines and chemokines involved in the pathogenesis of this disease.

Grau de incapacidade física na população idosa afetada pela hanseníase no estado da Bahia, Brasil / Physical disability degree in the elderly population affected by leprosy in the state of Bahia, Brazil

Objetivo: Analisar o grau de incapacidade física na população idosa afetada pela hanseníase no estado da Bahia, entre 2001 e 2012. Métodos: Os dados referentes aos casos de hanseníase foram obtidos do Sistema Nacional de Agravos de Notificação. Variáveis analisadas: sexo, faixa etária, raça/cor, escolaridade, classificação clínica e operacional, grau de incapacidade física no diagnóstico e na alta. Foram calculados indicadores epidemiológicos relacionados à incapacidade física. Resultados: A hanseníase apresenta elevada magnitude na população idosa, com coeficiente de detecção de casos novos superior ao da população geral, situando-se em nível hiperendêmico. Quanto ao perfil epidemiológico da hanseníase em idosos, destaca-se: homens, faixa etária 60 a 69 anos, raça branca, baixa escolaridade, forma clínica dimorfa e classificação operacional multibacilar. 36,25% dos casos diagnosticados apresentavam incapacidade física no momento do diagnóstico, com destaque para o gênero masculino. Conclusão: A elevada proporção de indivíduos com incapacidades físicas no momento do diagnóstico sugere diagnóstico tardio e prevalência oculta da doença, sobretudo em indivíduos do gênero masculino

Objective: To analyze the degree of physical disability in the elderly population affected by leprosy in Bahia State, between 2001 and 2012. Methods: The data relating to cases of leprosy was obtained from National System of Notifiable Diseases. Variables analyzed gender, age, race/color, education level, clinical and operational classification, degree of physical incapacity in diagnosis and discharge. Epidemiological indicators related to physical incapacity were calculated. Results: The leprosy features high magnitude in the elderly population, with a coefficient of detection of new cases higher than the general population, situated at a hyperendemic level. As to the epidemiological profile of leprosy in elderly, stands out: men, age 60 to 69 years, white race, low education level, dimorphic clinical manifestation and multibacillary operational classification. 36.25% of diagnosed cases had a physical incapacity at the time of diagnosis, with emphasis on the masculine gender. Conclusion: The high proportion of individuals with physical incapacity at the time of diagnosis suggests late diagnosis and hidden prevalence of the disease, mostly in Individuals of the male gender

Avaliação da produção de óxido nítrico, citocinas e expressão de receptores Toll em leucócitos do sangue periférico de pacientes com reações hansênicas e formas clínicas da hanseníase

O objetivo deste estudo foi identificar biomarcadores associados com as distintas formas/manifestações da hanseníase. Características fenotípicas (TLR2; TLR4; HLA-DR) e funcionais (NO-DAF-2T; IL-8;TNF-α; IL-4; IL-10;TGF-β) da imunidade inata (Neutrofilos-NEU; Monocitos-MON) e adaptiva (CD4+; CD8+T-cells) foram avaliadas ex vivo e sob estímulo de M. lepraein vitro (*). A população do estudo incluiu indivíduos com as forma Tuberculóide (DD/DT/T); Virchowiano (DV/V); reação hansênica Tipo-1; reação hansênica Tipo-2 e indivíduos saudáveis controle (NI). A análise dos dados demonstrou que independente da forma clínica, pacientes com hanseníase apresentaram baixo NEU TLR4+ e MON TGF-β+, enquanto DV/V apresentou alto MON HLA-DR+. Ambos DD/DT/T e DV/V apresentou aumento de CD4+IL-10+, mas não somente DV/V apresentou aumento de CD8+TLR2+ e IL-10. Pacientes com reação Tipo-1 exibiram, no geral, uma baixa na regulação de TLR na imunidade inata e adaptativa (NEU: MON TLR2+ e NEU: CD4+TLR4+).

Foi observada uma alta regulação de CD4+: CD8+IL-10+ nos indivíduos com reação hansênica, mas somente pacientes com reação do Tipo-2 mostrou aumento em MON IL-10+. Assinaturas de biomarcadores específicos para M. leprae (*) revelaram uma expansão das características fenotípicas e funcionais nos pacientes com hanseníase com aumento de *MON NO-DAF-2T+ em DD/DT/T e alta regulação de *NEU TLR2+ e *CD8+IL-10+ em DV/V. Foi evidenciado diminuição na expressão dos principais biomarcadores dos pacientes com reação hansênica particularmente na imunidade adaptativa. Um resumo dos dados trabalhados indicou que MON IL-10+ e *NEU TLR4+ podem ser considerados como biomarcadores universais da hanseníase juntamente com MON TGF-β+; *MON TNF-α+; *CD4+TGF-β+; *CD8+TLR2+; *CD8+TNF-α+; *CD8+IL-4+; *CD8+TGF-β+ como um ensaio clínico relacionado com as características da hanseníase e *CD4+ como biomarcadores de reação hansênica. MON HLA-DR+; *CD8+IL-10+ foram biomarcadores polo seletivo para DV/V, NEU TLR2+; NEU TLR4+; CD4+TLR4+; *NEU NO-DAF-2T+; *MON TLR2+; *MON HLA-DR+; *CD4+TLR2+; *CD4+TLR4+; *CD8+ para reação hansênica Tipo-1 e MONIL-10+; *CD8+IL-8+ para Tipo-2. Em conclusão, este estudo mostrou a existência de diferenças em marcadores imunológicas entre as formas/manifestações da hanseníase que poderão ser futuramente aplicados como biomarcadores para estudos clínicos.

Risk-benefit assessment of Bacillus Calmette-Guérin vaccination, anti-phenolic glycolipid I serology, and Mitsuda test response: 10-year follow-up of household contacts of leprosy patients

Abstract: INTRODUCTION: Despite multidrug therapy, leprosy remains a public health issue. The intradermal Bacillus Calmette-Guérin (BCG) vaccine, Mitsuda test (lepromin skin test), and anti-phenolic glycolipid I (PGL-I) serology are widely used in leprosy studies and have shown great epidemiological value. METHODS: This longitudinal study evaluated the relative risks and benefits of these three tools by comparing results observed in household contacts (HHCs) of leprosy patients who developed leprosy with those of HHCs who did not in a population of 2,992 individuals monitored during a 10-year period. RESULTS : Seventy-five (2.5%) new leprosy cases were diagnosed, including 28 (0.9%) co-prevalent cases. Therefore, for the risk-benefit assessment, 47 (1.6%) HHCs were considered as truly diagnosed during follow-up. The comparison between healthy and affected contacts demonstrated that not only did BCG vaccination increase protection, but boosters also increased to 95% relative risk (RR) reduction when results for having two or more scars were compared with having no scars [RR, 0.0459; 95% confidence interval (CI), 0.006-0.338]. Similarly, Mitsuda reactions >7mm in induration presented 7-fold greater protection against disease development compared to reactions of 0-3mm (RR, 0.1446; 95% CI, 0.0566-0.3696). In contrast, anti-PGL-I ELISA seropositivity indicated a 5-fold RR increase for disease outcome (RR, 5.688; 95% CI, 3.2412-9.9824). The combined effect of no BCG scars, Mitsuda reaction of <7mm, and seropositivity to anti-PGL-I increased the risk for leprosy onset 8-fold (RR, 8.109; 95% CI, 5.1167-12.8511). CONCLUSIONS: The adoption of these combined assays may impose measures for leprosy control strategies.

Análise da incidência da reação reversa em uma coorte de pacientes com hanseníase multibacilar tratados com 12 e 24 doses de poliquimioterapia / Analysis the incidence of reversal reaction in a cohort of multibacillary patients treated with 12 and 24 doses of multidrug therapy

A reação reversa (RR), uma complicação da hanseníase, ainda traz consigo muitos questionamentos, apesar dos estudos existentes. Em 1997, a Organização Mundial da Saúde (OMS) preconizou uma redução na duração do tratamento da hanseníase,passando de 24 doses para 12 doses fixas de poliquimioterapia (PQT), no entanto,poucos estudos foram realizados comparando os dois tratamentos sob a ótica desse quadro reacional. O objetivo do presente estudo foi avaliar a incidência da RR em pacientes com hanseníase multibacilar (MB) submetidos a 24 e 12 doses da PQT. A partir de um estudo observacional, longitudinal, retrospectivo, foram analisados osdados secundários de 888 pacientes classificados segundo Ridley e Jopling (1966)em borderline-borderline (BB), borderline-lepromatoso (BL) e lepromatoso (LL),atendidos no Ambulatório Souza Araújo (ASA) – Instituto Oswaldo Cruz (IOC), emum período de 25 anos. Destes, 261 pacientes apresentaram episódios de RR,sendo 112 no grupo de 24 doses e 149 no de 12 doses. Os grupos também foram analisados quanto às características sócio-demográficas e clínicas, tendo sido feitasanálises descritivas com distribuição de freqüências para todas as variáveis incluídas no estudo. Objetivando comparar os grupos utilizamos os testes quiquadrado(variáveis categóricas) e t de Student (variáveis contínuas).Observamos uma maior incidência da RR (20,3 episódios/1000 pessoas/mês) nogrupo de 12 doses PQT...

Reversal reactions episodes, an complication of leprosy, still brings questions despitethe existence of several studies. In mid-1997, the World Health Organizationrecomended a reduction on leprosy treatment duration from 24 doses to 12 fixeddoses of multidrug therapy, however, a few studies has been made comparing bothtreatments by the reversal reaction perspective. The aim of this study was toevaluate the incidence of RR in patients with multibacillary (MB) disease submittedon 24 and 12 doses of MDT.From an observational, longitudinal and retrospective study, secondary data of 888patients classified accordding to Ridley Jopling`s (1966) system of classification inborderline-borderline (BB), borderline-lepromatous (BL) and lepromatous (LL)forms, treated at Ambulatório Souza Araújo (ASA) – Oswaldo Cruz Institute (IOC) ina 25 years period, submitted to MDT 24 and 12 doses were analyzed. Of these, 261patients had episodes of reversal reaction, with 112 patients in group of 24 dosesand 149 in group 12 doses. These groups were also analyzed as to sociodemographicand clinical characteristics, and a descriptive analysis was made withfrequency distribution for all variables included in this study. In order to compare thegroups, we used the chi-square statistic (categoric variable) and t Student statistic(continuous variable)...

Envolvimento do receptor TLR-9 na resposta imune do hospedeiro durante o curso da infecção pelo Mycobacterium leprae / Involvement of the TLR9 receptor in host immune responses during infection by Mycobacterium leprae

Mycobacterium leprae, agente etiológico da hanseníase, expressa em abundância uma proteína catiônica semelhante às histonas, denominada Hlp, presente tanto no envelope como no nucleóide bacteriano. O reconhecimento do DNA bacteriano, rico em motivos CpG não metilados, pelo receptor TLR-9 representa uma importante via para a ativação da resposta imune inata, a qual pode levar à eliminação do agente infeccioso ou mediar manifestações patológicas. Foi mostrado ainda que complexos DNA-histona são mais potentes agonistas de TLR-9 que DNA sozinho. Assim, o presente trabalho teve como objetivo investigar o envolvimento do receptor TLR-9 na ativação da resposta imune do hospedeiro durante o curso da infecção pelo M. leprae.Inicialmente foi analisada a participação do TLR-9 na ativação da resposta imune inata em células epiteliais alveolares da linhagem A549 após estímulo com M. leprae. M. leprae foi capaz de induzir aumento das quimiocinas IL-8 eMCP-1 e a transcrição gênica do peptídeo antimicrobiano HbetaD-2 nas células epiteliais. O aumento da expressão de CD80 na superfície celular também foi observada após estímulo com o bacilo. O complexo CpG-Hlp micobacteriano solúvel também induziu aumento na produção de IL-8 nas células A549. Foi observado que o aumento de IL-8 induzido pelo M. leprae ocorre de forma dependente da translocação nuclear do NF-capaB e que o antagonista sintético deTLR-9 afetou a secreção de IL-8 induzida pelo M. leprae. A adição de CpG aoM. smegmatis selvagem, mas não mutante para o gene hlp, aumentou a produção de IL-8 pelas células epiteliais. Em conjunto, esses dados sugerem que as células epiteliais respiratórias podem reconhecer M. leprae via TLR-9 e,assim, participar da resposta imune inata no sítio inicial da infecção. Uma vez que o aparecimento do eritema nodoso hansênico (ENH) está associado a liberação massiva de antígenos micobacterianos, foi investigado o envolvimento do TLR-9 na patogênese do ENH...

Mycobacterium leprae, etiological agent of leprosy, expresses in abundance acationic protein similar to histones, called histone-like protein (Hlp), present inthe envelope as well as in bacterial nucleoid.The recognition of bacterial DNArich in unmethylated CpG motifs by TLR-9 is an important pathway for activationof the innate immune response, which can lead to the elimination of theinfectious agent or mediate pathological manifestations. Moreover, studiesshowed that DNA-histone complexes are more potent agonists of TLR-9 thanDNA alone. This study aimed to investigate the involvement of TLR-9 in theactivation of the host immune response during the course of M. leprae infection.Initially, we analyzed the participation of TLR-9 activation on the innate immuneresponse in A549 alveolar epithelial cells after stimulation with M. leprae. It wasshown that M. leprae was able to induce the chemokines IL-8 and MCP-1, andgene transcription of antimicrobial peptide HbetaD-2 in epithelial cells. Theincrease of CD80 expression on the cell surface was also observed afterstimulation with bacillus. Soluble mycobacterial CpG-Hlp complex also inducedan increase in IL-8 in A549 cells. It was observed that the increase of IL-8,induced by M. leprae, occurs dependently nuclear translocation of NF-capaB andsynthetic TLR-9 antagonist affected IL-8 secretion induced by M. leprae. Theaddition of CpG to wild type M. smegmatis, but not to the mutant gene hlp,increased IL-8 production by epithelial cells. As a whole, these results suggestthat respiratory epithelial cells can recognize M. leprae via TLR-9 and thusparticipate in the innate immune response in the initial infection site. Since theappearance of erythema nodosum leprosum (ENL) is associated with themassive release of mycobacterial antigens, it was investigated the involvementof TLR-9 in the pathogenesis of ENL...

Serology with ML Flow test in health professionals from three different states of Brazil / Sorologia com o teste ML Flow em profissionais de saude em tres diferentes estados do Brasil

BACKGROUND: In highly endemic countries, transmission and sub-clinical infection of leprosy are likely and the disease manifests itself in individuals without any known close contact with a leprosy patient. Health workers are social contacts belonging to the same network (the Health System) and some of them share the same social environment (nursing assistants) as patients with known patients and / or carriers. OBJECTIVE: To identify ML Flow seropositivity among health professionals. METHODS: We conducted a cross-sectional study using a serological survey with the ML Flow test in 450 health professionals (doctors, nurses and nursing assistants), in order to detect seropositivity in areas of high and low endemicity in municipalities from three Brazilian states (RJ, MS and RS). RESULTS: The results showed general 16% seropositivity, higher in low endemic areas, regardless of whether there was direct care for leprosy patients. Paradoxically, a statistical association was observed between the area studied and seropositivity, as the place with the lowest endemicity (CA) had the highest seropositivity rate (p = 0.033). CONCLUSION: The authors suggest these results are associated with a presence of an unspecified link to bovine serum albumin (BSA), carrier of PGL-1 in the ML Flow test, and recommend expanded seroepidemiological research utilizing tests with human and bovine albumin. .

FUNDAMENTOS: Em países altamente endêmicos a transmissão e infecção sub-clínica da hanseníase provavelmente ocorrem e a doença se manifesta em indivíduos sem qualquer contato próximo conhecido com paciente com hanseníase. Os trabalhadores de saúde são contatos sociais que pertencem à mesma rede (Sistema de Saúde) e alguns deles compartilham o mesmo ambiente social (auxiliares de enfermagem) com pacientes conhecidos e/ou portadores. OBJETIVO: Conhecer a soropositividade ao ML Flow entre os profissionais de saúde. MÉTODOS: Foi realizado um estudo transversal através de inquérito sorológico com o teste ML Flow em 450 profissionais de saúde (médicos, enfermeiros e auxiliares de enfermagem) visando conhecer a soropositividade em áreas de alta e baixa endemicidade em municípios de três estados brasileiros (RS, MS e RJ). RESULTADOS: Os resultados mostraram 16% de soropositividade em geral, mais elevada na área de baixa endemicidade, independente da assistência direta a pacientes com hanseníase. Paradoxalmente foi observada associação estatística entre a área estudada e soropositividade, apontando o lugar de mais baixa endemicidade (CA) com o maior valor (p=0,033). CONCLUSÃO: os autores sugerem a presença de ligação inespecífica a soroalbumina bovina (BSA), carreadora do antígeno PGL-1 no teste ML Flow para explicar os resultados inesperados e recomendam testagem ampliada utilizando testes com albumina humana e bovina. .

Identification of clinical, epidemiological and laboratory risk factors for leprosy reactions during and after multidrug therapy

This cross-sectional retrospective study evaluated 440 leprosy patients; 57% (251/440) had leprosy reactions during and/or after multidrug therapy, 80.5% (202/251) of whom presented with multibacillary leprosy. At diagnosis, positive bacterial index (BI) [odds ratio (OR) = 6.39; 95% confidence interval (CI): 4.1-10.1)] or polymerase chain reaction (PCR) (OR = 9.15; 95% CI: 5.4-15.5) in skin smears, anti-phenolic glycolipid-1 (anti-PGL-1) ELISA (OR = 4.77; 95% CI: 2.9-7.9), leucocytosis (OR = 9.97; 95% CI: 3.9-25.7), thrombocytopenia (OR = 5.72; 95% CI: 2.3-14.0) and elevated lactate dehydrogenase (OR = 2.38; 95% CI: 1.4-4.0) were potential markers for the development of reactions during treatment. After treatment, positive BI (OR = 8.47; 95% CI: 4.7-15.3) and PCR (OR = 6.46; 95% CI: 3.4-12.3) in skin smears, anti-PGL-1 ELISA (OR = 2.25; 95% CI: 1.3-3.9), anaemia (OR = 2.36; 95% CI: 1.2-4.5), leucocytosis (OR = 4.14; 95% CI: 1.5-11.6) and thrombocytopenia (OR = 3.70; 95% CI: 1.3-2.2) were risk factors for the occurrence of reactions during the study period. The identification of groups with an increased risk for developing reactions will allow for the timely development of a treatment plan to prevent nerve damage and, therefore, the appearance of the disabling sequelae associated with the stigma of leprosy.

Oral coinfection can stress peripheral lymphocyte to inflammatory activity in leprosy

INTRODUCTION: This study evaluated the intracellular profile of interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-10 (IL-10) and interferon-γ (IFN-γ) in peripheral blood mononuclear cells (PBMCs) from leprosy patients based on oral infections presence to determine whether these coinfections could be associated with pro-inflammatory activity in leprosy. METHODS:Leprosy patients regardless of clinical form and specific leprosy treatment (n=38) were divided into two groups: Group I - leprosy patients with oral infections (n=19), and Group II - leprosy patients without oral infections (n=19). Non-leprosy patients presenting oral infections were assigned to the control Group (n=10). Intracellular IL-2, IL-4, IL-10 and IFN-γ production was evaluated by flow cytometry (FACS) before and 7 days after controlling the oral infection in the Group I, before and 7 days after dental prophylaxis in the Group II, and during oral infection process in control Group. RESULTS: Low percentages of CD3+ lymphocytes bearing IL-2, IL-10 and IFN-γ were observed in the Group I and Group II at baseline and 7 days after therapy or prophylaxis compared to controls. Group I showed reduced percentages of IL-4 at baseline and 7 days after therapy compared to controls, or at baseline of Group II, and the Group II showed reduced percentages of CD3+ cells bearing IL-4 compared to control. An increase of the percentages of CD3+cells bearing IL-4 was observed in the Group I after the oral infections treatment. CONCLUSIONS: The occurrence of oral infections favors the intracellular cytokines expression and, probably, the inflammatory reaction operating as a stimulatory signal triggering the leprosy reactions.

Episódios reacionais hansênicos: estudo de fatores relacionados com adesão ao tratamento em uma unidade de referência / Leprosy rectional episodes: study of factores related to adherence to treatment in a unit of reference

As reações hansênicas são um dos maiores problemas para os profissionais de saúde no manejo dos portadores de hanseníase, tornando o controle das mesmas de fundamental importância para evitar complicações clínicas e sociais. A assiduidade às consultas para avaliação e controle é essencial para a cura. Este estudo avaliou a influência dos fatores sócio-demográficos e clínicos na adesão ao tratamento em estado reacional em um ambulatório de referência. Trata-sede um estudo do tipo série de casos, em pacientes em controle no ambulatório do NMT/UFPA nos anos de 2008 a 2009. Os resultados mostraram que pacientes do sexo masculino, adultos e com baixa escolaridade foram os mais acometidos pela hanseníase. As reações hansênicas do tipo 1(RR) foram as mais incidentes, tendo como principal queixa a dor. Apesar dos resultados não terem mostrado associação, com significância estatística, entre as variáveis estudadas e a não adesão ao tratamento, eles evidenciaram variáveis que interferem na assiduidade às consultas.

Leprosy reactions are one of the biggest problems for health professionals in the management of patients with leprosy, making control of these fundamentally important to avoid medical complications and social problems. The attendance to appointments for evaluation and control is essential to healing. This study evaluated the influence of socio-demographic and clinical factors in adherence to treatment in a reactive state in a outpatient referral. This is a study of type series of cases in patients in control in the outpatient department of the NMT / UFPA in the years 2008-2009. The results showed that male, adults and low education patients were the most affected by leprosy. Leprosy reactions of 1 (RR) type had the highest incidence, with pain as the chief complaint. Although the results have not shown an association with statistical significance between variables and non-adherence to treatment, they showed variables that interfere in attendance to appointments.

Epidemiologia genética em hanseníase: estudo de associação da região genômica candidata 6p21 e do gene TLR1 / Genetic epidemiology of leprosy: association study of the genomic region 6p21 canditate and TLR1 gene

A hanseníase é uma doença infecciosa crônica, que acomete pele e sistema nervoso periférico e tem como agente etiológico o Mycobacterium leprae, um patógeno exclusivamente intracelular, que tem predileção por macrófagos e pelas células de Schwann. É um traço complexo e fatores genéticos do hospedeiro têm sido repetidamente implicados com o risco para a doença. A região cromossômica 6p21 vem sendo sistematicamente envolvida com a hanseníase, não só pelos genes do HLA de classe II, como também pelos estudos envolvendo marcadores em genes como o TNF e a LTA. O gene TLR1 também é um importante candidato e polimorfismos deste já têm sido associados com hanseníase per se e com reação hansênica. O objetivo desta pesquisa foi conduzir estudo de associação de base populacional do tipo caso-controle em hanseníase testando marcadores do tipo tag SNPs em genes candidatos da região cromossômica candidata 6p21 e do gene TLR1. Oitenta e nove marcadores do tipo tag SNPs, localizados em trinta e seis genes foram genotipados. O presente trabalho envolveu 1718 indivíduos, 981 casos e 737 controles, provenientes de dois estados brasileiros: Mato Grosso e São Paulo. As genotipagens da população de Rondonópolis, MT foram realizadas em plataforma de médio rendimento (VeraCode GoldenGate Genotyping Assay – Illumina) e as genotipagens da população de São Paulo foram feitas usando discriminação alélica baseada na tecnologia TaqMan (Applied Biosystems). Para as análises estatísticas foi empregado modelo de regressão logística, com correção para as co-variáveis etnia e sexo, usando o software R, para Windows. Treze genes localizados na região 6p21 tiveram marcadores associados com hanseníase per se. O alelo S do polimorfismo N248S do gene TLR1 também foi associado com susceptibilidade para hanseníase per se...

Leprosy is an chronic infectious disease that attacks skin and peripheral nervous system. The causative agent is Mycobacterium leprae, an obligate intracellular pathogen that infects macrophage and Schwann cells. It is a complex trait and host genetic factors have been extensively implicated in leprosy susceptibility. The chromosomal region 6p21 has been involved with leprosy susceptibility due to HLA class II, and TNF and LTA genes, as well. The TLR1 gene is also an important candidate gene and polymorphisms at this locus have been associated to leprosy per se and leprosy reactions. This research is a population-based association study in leprosy which tested tag SNPs located at candidate genes in chromosomal region 6p21 and in TLR1 gene. Eighty-nine markers distributed in thirty-six genes were genotyped. The present work enrolled 1,718 individuals, 981 cases and 737 controls from Mato Grosso and São Paulo States, Brazil. The genotypes for Rondonópolis population were obtained using by medium-scale genotyping platform (VeraCode GoldenGate Genotyping Assay – Illumina), while to São Paulo samples the genotyping were done by allelic discrimination based on TaqMan technology (Applied Biosystems). Statistical analysis were performed by logistic regression models adjusted for the covariates sex and ethnicity, using R software. Thirteen genes located at 6p21 region presented markers associated to leprosy per se. The S allele for N248S polymorphism at TLR1gene was also associated to leprosy susceptibility...

Resposta imune celular Mycobacterium leprae-específica in vitro de contatos de pacientes com hanseníase multibacilar no início e durante o tratamento do caso índice / Cellular immune response Mycobacterium leprae-specific in vitro contact multibacillary leprosy patients at the beginning and during the treatment of the index case

Os indivíduos com hanseníase multibacilar (MB) apresentam uma baixa capacidade de produzir IFN-gama em resposta ao Mycobacterium leprae, agente etiológico da doença, e consequentemente, apresentam uma alta carga bacilar constituindo as principais fontes de disseminação e transmissão do bacilo. [...] O presente estudo tem como objetivo geral compreender os mecanismos de modulação negativa da resposta imune celular M. leprae específica em indivíduos sadios com alta exposição ao bacilo e em indivíduos com hanseníase MB, até hoje pouco esclarecidos. Para isto, avaliamos as frequências de linfócitos T CD4+ e CD8+ virgens, de memória central e de memória efetora recém-ativados em resposta ao M. leprae ou conjuntos de peptídeos nonâmeros (restritos a MHC de classe I, P.C.I) e pentadecâmeros (restritos a MHC de classe II, P.C.II) específicos do M. leprae em células mononucleares de sangue periférico (CMS) dos contatos domiciliares no início (T0) e ao longo do tratamento do caso índice (T1). Em paralelo, os níveis de 16 citocinas, quimiocinas e fatores de crescimento secretados por CMS destes contatos estimuladas in vitro, em T0 e T1, com os antígenos mencionados. Nove contatos foram avaliados, dos quais sete foram vacinados com BCG em T0. Na maioria dos contatos, independente da vacinação ou não com BCG, observou-se um aumento da frequência de linfócitos T CD4+ de memória efetora a pelo menos um dos antígenos utilizados em T1. Detectou-se, também, um aumento consistente de células T CD8+ de memória efetora respondedoras a P.C.I. Observou-se variações nas frequências de linfócitos T CD4+ e/ou CD8+ produtores de IFN-gama ou IL-10 M. leprae específicas no sangue periférico dos contatos...

Individuals with multibacillary leprosy (MB) have a low capacity to produce IFN-gama in response to Mycobacterium leprae, etiological agent of leprosy, and consequently, have a high bacterial load, being the main sources of dissemination and transmission of thebacillus. [...] The aim of the present study is to understand the mechanismsof cellular immune response downregulation by M. leprae in healthy individuals with high exposure to M. leprae and patients with MB leprosy, a phenomenon that remains poorly understood until today. In this work we evaluated the frequencies of recentlyactivated naive, central memory and effector memory CD4+ and CD8+T cells, in response to M. leprae and a pool of M. leprae-specific peptide nonamers (MHC class Irestricted,P.C.I) and pentadecamers (MHC class II-restricted, P.C.II) in the peripheralblood mononuclear cells (PBMC) of HC at the beginning (T0) and during the treatment of the index case (T1). We also compared the levels of 16 cytokines, chemokines andgrowth factors secreted by PBMC of these contacts stimulated in vitro with the same antigens at T0 and T1. Nine contacts were evaluated, which seven were vaccinated with BCG at T0. Most of the HC, independently of BCG vaccination, showed increasedfrequency of CD4+ effector memory to at least one of the antigens used in T1. A consistent increase of CD8+effector memory responsive to P.C.I was also detected. It was observed variations in the frequencies of CD4+and/or CD8+ T cells producing IFN-gama or IL-10 M. leprae-specific by PBMC from HC...

Differential expression of the costimulatory molecules CD86, CD28, CD152 and PD-1 correlates with the host-parasite outcome in leprosy

Leprosy is a spectral disease exhibiting two polar sides, namely, lepromatous leprosy (LL) characterised by impaired T-cell responses and tuberculoid leprosy in which T-cell responses are strong. Proper T-cell activation requires signalling through costimulatory molecules expressed by antigen presenting cells and their ligands on T-cells. We studied the influence of costimulatory molecules on the immune responses of subjects along the leprosy spectrum. The expression of the costimulatory molecules was evaluated in in vitro-stimulated peripheral blood mononuclear cells of lepromatous and tuberculoid patients and healthy exposed individuals (contacts). We show that LL patients have defective monocyte CD86 expression, which likely contributes to the impairment of the antigen presentation process and to patients anergy. Accordingly, CD86 but not CD80 blockade inhibited the lymphoproliferative response to Mycobacterium leprae. Consistent with the LL anergy, there was reduced expression of the positive signalling costimulatory molecules CD28 and CD86 on the T-cells in these patients. In contrast, tuberculoid leprosy patients displayed increased expression of the negative signalling molecules CD152 and programmed death-1 (PD-1), which represents a probable means of modulating an exacerbated immune response and avoiding immunopathology. Notably, the contacts exhibited proper CD86 and CD28 expression but not exacerbated CD152 or PD-1 expression, suggesting that they tend to develop a balanced immunity without requiring immunosuppressive costimulatory signalling.

Leprosy, a neglected disease that causes a wide variety of clinical conditions in tropical countries

Leprosy is an ancient disease that remains endemic and continues to be a major public health problem in some tropical countries, where it has been internationally recognized as being linked to the underdevelopment conditions. The natural course of the disease covers a wide variety of clinical conditions with systemic involvement. In this paper, we review the findings obtained in studies of the pathological mechanisms of leprosy, including a survey of the literature and of our own work. The understanding and control of the wide variety of clinical conditions should help improve patient care and thus prevent the onset of physical impairment and the stigma of the disease.

Comparison of two rapid tests for anti-phenolic glycolipid-I serology in Brazil and Nepal

The diagnosis of leprosy continues to be based on clinical symptoms and early diagnosis and treatment are critical to preventing disability and transmission. Sensitive and specific laboratory tests are not available for diagnosing leprosy. Despite the limited applicability of anti-phenolic glycolipid-I (PGL-I) serology for diagnosis, it has been suggested as an additional tool to classify leprosy patients (LPs) for treatment purposes. Two formats of rapid tests to detect anti-PGL-I antibodies [ML immunochromatography assay (ICA) and ML Flow] were compared in different groups, multibacillary patients, paucibacillary patients, household contacts and healthy controls in Brazil and Nepal. High ML Flow intra-test concordance was observed and low to moderate agreement between the results of ML ICA and ML Flow tests on the serum of LPs was observed. LPs were "seroclassified" according to the results of these tests and the seroclassification was compared to other currently used classification systems: the World Health Organization operational classification, the bacilloscopic index and the Ridley-Jopling classification. When analysing the usefulness of these tests in the operational classification of PB and MB leprosy for treatment and follow-up purposes, the ML Flow test was the best point-of-care test for subjects in Nepal and despite the need for sample dilution, the ML ICA test yielded better performance among Brazilian subjects. Our results identified possible ways to improve the performance of both tests.

Genetics of leprosy reactions: an overview

Type-1 (T1R) and Type-2 (T2R) leprosy reactions (LR), which affect up to 50% of leprosy patients, are aggressive inflammatory episodes of sudden onset and highly variable incidence across populations. LR are often diagnosed concurrently with leprosy, but more frequently occur several months after treatment onset. It is not uncommon for leprosy patients to develop recurring reactional episodes; however, they rarely undergo both types of LR. Today, LR are the main cause of permanent disabilities associated with leprosy and represent a major challenge in the clinical management of leprosy patients. Although progress has been made in understanding the immunopathology of LR, the factors that cause a leprosy patient to suffer from LR are largely unknown. Given the impact that ethnic background has on the risk of developing LR, host genetic factors have long been suspected of contributing to LR. Indeed, polymorphisms in seven genes [Toll-like receptors (TLR)1, TLR2, nucleotide-binding oligomerisation domain containing 2, vitamin D receptor, natural resistance-associated macrophage protein 1, C4B and interleukin-6] have been found to be associated with one or more LR outcomes. The identification of host genetic markers with predictive value for LR would have a major impact on nerve damage control in leprosy. In this review, we present the recent advances achieved through genetic studies of LR.