Association between NDO-LID and PGL-1 for leprosy and class I and II human leukocyte antigen alleles in an indigenous community in Southwest Amazon

The frequencies of the Human leukocyte antigen (HLA) alleles in the Puyanawa indigenous reserve population and their association with the NDO-LID and ELISA PGL-1 rapid serological test was assessed. This was a cross-sectional study with an epidemiological clinical design conducted in two indigenous communities in the state of Acre, Brazil. Blood was collected in a tube with EDTA to identify HLA alleles and perform serological tests. DNA was obtained using the salting out procedure. The LabType™ technique (One-Lambda-USA) was used for HLA class I (loci A*, B* and C*) and II (loci DRB1*, DQA1* and DQB1*) typing. Allele frequency was obtained by direct count, and the chi-square test was used to assess the association with the NDO-LID and PGL-1 tests. The most frequent alleles in the two communities were: HLA-A*02:01, HLA-B*40:02, HLA-DRB1*16:02, HLA-DQA1*05:05 and HLA-DQB1*03:01. The allele HLA-C*04:01 was the most common in the Barão community, and the allele HLA-C*07:01 in Ipiranga. Among individuals who presented seropositivity to the NDO-LID test, the association with alleles HLA-A*02 (43.18% vs 24.8%, p = 0.03, OR = 2.35) and HLA-B*53 (6.83% vs 0.0%, p = 0.03, OR = 8.95) was observed in the Barão community. HLA-B*15 was associated with non-seroconversion to the NDO-LID test in Ipiranga. In both communities, HLA-B*40 and HLA-C*03 were associated with positive serological response to ELISA PGL-1. The HLA class I and II alleles most frequently found in this study have already been described among Terena indigenous groups, and HLA class I contributes to seroconversion to NDO-LID and PGL-1 tests in inhabitants of the Barão and Ipiranga communities(AU).

Association between NDO-LID and PGL-1 for leprosy and class I and II human leukocyte antigen alleles in an indigenous community in Southwest Amazon.

The frequencies of the Human leukocyte antigen (HLA) alleles in the Puyanawa indigenous reserve population and their association with the NDO-LID and ELISA PGL-1 rapid serological test was assessed. This was a cross-sectional study with an epidemiological clinical design conducted in two indigenous communities in the state of Acre, Brazil. Blood was collected in a tube with EDTA to identify HLA alleles and perform serological tests. DNA was obtained using the salting out procedure. The LabType™ technique (One-Lambda-USA) was used for HLA class I (loci A*, B* and C*) and II (loci DRB1*, DQA1* and DQB1*) typing. Allele frequency was obtained by direct count, and the chi-square test was used to assess the association with the NDO-LID and PGL-1 tests. The most frequent alleles in the two communities were: HLA-A*02:01, HLA-B*40:02, HLA-DRB1*16:02, HLA-DQA1*05:05 and HLA-DQB1*03:01. The allele HLA-C*04:01 was the most common in the Barão community, and the allele HLA-C*07:01 in Ipiranga. Among individuals who presented seropositivity to the NDO-LID test, the association with alleles HLA-A*02 (43.18% vs 24.8%, p=0.03, OR=2.35) and HLA-B*53 (6.83% vs 0.0%, p=0.03, OR=8.95) was observed in the Barão community. HLA-B*15 was associated with non-seroconversion to the NDO-LID test in Ipiranga. In both communities, HLA-B*40 and HLA-C*03 were associated with positive serological response to ELISA PGL-1. The HLA class I and II alleles most frequently found in this study have already been described among Terena indigenous groups, and HLA class I contributes to seroconversion to NDO-LID and PGL-1 tests in inhabitants of the Barão and Ipiranga communities.

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

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.

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.

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.

Factors associated with leprosy in children contacts of notified adults in an endemic region of Midwest Brazil

OBJECTIVES: To analyze the factors associated with leprosy in children who were intradomiciliary contacts of notified adults with the disease in an endemic municipality in Mato Grosso, Brazil. METHOD: Case-control study with 204 children under 15 years of age, living in an endemic municipality. Cases (n=40) were considered as the children with leprosy registered at the National Information System of Notifiable Diseases in 2014 and 2015, who were intradomiciliary contacts of at least one adult diagnosed with the disease in the family, and as a control group (n=164) of children living within a radius of up to 100m of the notified cases. Data were obtained through medical file analysis, interviews, and blood samples for anti-PGL-I serological test by the ELISA method. The binary logistic regression technique was used, with p≤0.05. RESULTS: After adjustments, the following were associated with leprosy age (95% CI 1.24-9.39, p=0.018), area of residence (95% CI 1.11-6.09, p=0.027), waste disposal (95% CI 1.91-27.98, p=0.004), family history of the disease (95% CI 3.41-22.50, p=0.000), and time of residence (95% CI 1.45-7.78, p=0.005). CONCLUSION: Factors associated with the disease indicate greater vulnerability of children aged 8-14 years, associated with living conditions and time of residence, as well as the family history of the disease.

Can anti-PGL-1 and anti-NDO-LID-1 antibody titers be used to predict the risk of reactions in leprosy patients?

Leprosy patients may present reactional episodes classified as type I or reversal reaction and type II or erythema nodosum leprosum. Early diagnosis of these reactions is hampered by lack of diagnostic tests. This study aimed at evaluating anti-Mycobacterium leprae antibody levels in reactional and nonreactional leprosy patients at the time of diagnosis. Clinical data and serum samples of 224 patients diagnosed between 2009 and 2010 were collected in the municipality of Rondonópolis-MTBR. Quantification of anti-phenolic glycolipid-1 (PGL-1) IgM antibodies of M. leprae was obtained by the enzyme-linked immunosorbent assay method and anti-natural octyl disacharide-leprosy IDRI diagnostic (NDO-LID-1) IgM/IgG semiquantitative rapid test. We obtained low serological levels of anti-PGL-1 and anti-NDO-LID-1 for tuberculoid (T) (1.56% and 15.62%) and borderline tuberculoid (BT) patients (7.95% and 26.13%), medium levels in the borderline-borderline (BB) (47.91% and 68.75%), and high levels in lepromatous (LL) (93.33% and 100%) and borderline-lepromatous (BL) (88.0% and 100%). When comparing the reactional groups (RI and RII) with without reaction (WR) group at the time of diagnosis, we observed a statistically significant difference between the groups; patients with RII presented higher serological response: 66.66% anti-PGL-1 and 91.66% anti-NDO-LID-1. In respect to patients who developed a reaction after the initial diagnosis, they also showed significant positivity for both anti-PGL-1 and anti-NDO-LID-1 in comparison to the patients who stayed without reaction in the study period (P<0.0001). These results allow us to conclude that serological tests may contribute to an early diagnosis of RII and that the anti-NDO-LID-1 test was demonstrated to be a better indicator.

Can anti-PGL-1 and anti-NDO-LID-1 antibody titers to predict the risk of reactions in leprosy patients?

Leprosy patients may present reactional episodes classified as type I or reversal reaction and type II or erythema nodosum leprosum. Early diagnosis of these reactions is hampered by lack of diagnostic tests. This study aimed at evaluating anti­Mycobacterium leprae antibody levels in reactional and nonreactional leprosy patients at the time of diagnosis. Clinical data and serum samples of 224 patients diagnosed between 2009 and 2010 were collected in the municipality of Rondonópolis-MTBR. Quantification of anti­phenolic glycolipid-1 (PGL-1) IgM antibodies of M. leprae was obtained by the enzyme-linked immunosorbent assay method and anti­natural octyl disacharide-leprosy IDRI diagnostic (NDO-LID-1) IgM/IgG semiquantitative rapid test. We obtained low serological levels of anti­PGL-1 and anti­NDO-LID-1 for tuberculoid (T) (1.56% and 15.62%) and borderline tuberculoid (BT) patients (7.95% and 26.13%), medium levels in the borderline-borderline (BB) (47.91% and 68.75%), and high levels in lepromatous (LL) (93.33% and 100%) and borderline-lepromatous (BL) (88.0% and 100%). When comparing the reactional groups (RI and RII) with without reaction (WR) group at the time of diagnosis, we observed a statistically significant difference between the groups; patients with RII presented higher serological response: 66.66% anti­PGL-1 and 91.66% anti­NDO-LID-1. In respect to patients who developed a reaction after the initial diagnosis, they also showed significant positivity for both anti­PGL-1 and anti­NDO-LID-1 in comparison to the patients who stayed without reaction in the study period (P < 0.0001). These results allow us to conclude that serological tests may contribute to an early diagnosis of RII and that the anti­NDO-LID-1 test was demonstrated to be a better indicator.

Simultaneous analysis of multiple T helper subsets in leprosy reveals distinct patterns of Th1, Th2, Th17 and Tregs markers expression in clinical forms and reactional events.

Leprosy is a chronic infectious disease caused by Mycobacterium leprae. Previous studies have demonstrated that the difference among clinical forms of leprosy can be associated with the immune response of patients, mainly by T helper (Th) and regulatory T cells (Tregs). Then, aiming at clarifying the immune response, the expression of cytokines related to Th1, Th2, Th17 and Tregs profiles were evaluated by qPCR in 87 skin biopsies from leprosy patients. Additionally, cytokines and anti-PGL-1 antibodies were determined in serum by ELISA. The results showed that the expression of various targets (mRNA) related to Th1, Th2, Th17 and Tregs were significantly modulated in leprosy when compared with healthy individuals, suggesting the presence of a mixed profile. In addition, the targets related to Th1 predominated in the tuberculoid pole and side and Th2 and Tregs predominated in the lepromatous pole and side; however, Th17 targets showed a mixed profile. Concerning reactional events, Tregs markers were decreased and IL-15 was increased in reversal reaction and IL-17F, CCL20 and IL-8 in erythema nodosum leprosum, when compared with the respective non-reactional leprosy patients. Additionally, ELISA analysis demonstrated that IL-22, IL-6, IL-10 and anti-PGL-1 antibody levels were significantly higher in the serum of patients when compared with healthy individuals, and IL-10 and anti-PGL-1 antibodies were also increased in the lepromatous pole and side. Together, these results indicate that Th1, Th2 and Th17 are involved in the determination of clinical forms of leprosy and suggest that decreased Tregs activity may be involved in the pathogenesis of reactional events.

Detecção do risco de desenvolver reação mediante avaliação dos niveis de anticorpos anti-PGL-1 e anti-NDO-LID-1 em amostras de soros de pacientes com hanseniase, na vigência e na ausência de episódios reacionais / Detection of the risk of developing reaction by evaluating levels of anti-PGL-1 and Anti-NDO-LID-1 antibodies, in serum samples of leprosy patients, in the effectiveness and in the absence of reactive episodes

Pacientes com hanseníase podem apresentar surtos reacionais (reação tipo I e reação tipo II). Métodos sorológicos são desenvolvidos visando obter marcadores laboratoriais que auxiliem o diagnóstico precoce destes episódios. Diante disto, o objetivo deste estudo foi avaliar níveis de anticorpos de pacientes com hanseníase no momento do diagnóstico, com ou sem reação, e associar os resultados obtidos com tais fenômenos. Para isto, foram obtidos dados clínicos de prontuários e amostras séricas de 224 pacientes, diagnosticados no período de 2009/2010, no município de Rondonópolis-MT. Foram realizadas quantificações de anticorpos IgM anti-PGL-1 do Mycobacterium leprae (M. leprae) pelo método de ELISA, e detecção semiquantitativa de IgG/IgM anti-NDO-LID-1 pelo teste rápido. Obtivemos baixos níveis sorológicos de anti-PGL-1 e anti-NDO-LID-1 para os pacientes tuberculóides (T) (1,56% - 15,62%) e dimorfos-tuberculóides (DT) (7,95% - 26,13%), níveis médios nos dimorfos-dimorfos (DD) (47,91% - 68,75%) e elevados nos virchovianos (V) (93,33% - 100%) e dimorfos-virchovianos (DV) (88,88% - 100%). Ao compararmos os grupos reacionais (RI e RII) com os sem reação (SR) no momento do diagnóstico, observamos uma diferença estatística significativa entre os grupos, sendo que os pacientes com RII apresentaram maior resposta sorológica para ambos testes (anti-PGL-1 66,66% e anti-NDO-LID-1 91,66%). Os pacientes que vieram a desenvolver reação após o diagnóstico inicial, também apresentaram significativa positividade aos dois testes em comparação com aqueles que permaneceram sem reação no período estudado (p<0,0001). Estes resultados permitem concluir que os testes sorológicos podem contribuir para um diagnóstico precoce de RII, sendo que o teste anti-NDO-LID-1 demonstrou ser melhor indicador.

Patients with leprosy may present reactional outbreaks which are classified as Type I or Reverse Reaction and Type II or Nodose Hansenic Erythema. Serological methods have been developed aiming to obtain data that subsidize clinical and laboratory markers for the early diagnosis of these episodes. Therefore, the objective of this study was to evaluate antibody levels of leprosy patients at the time of diagnosis, with or without reactional episodes and to associate the results obtained with such phenomena. For this, clinical data of medical records and serum samples of 224 patients, diagnosed in the 2009/2010 period, were obtained in the municipality of Rondonópolis-MT. Quantifications of anti-PGL-1 IgM antibodies of Mycobacterium leprae (M. leprae) were performed by the ELISA method, and semiquantitative detection of anti-NDO-LID-1 IgM/IgG by the rapid test. We obtained low serological levels of anti-PGL-1 and anti-NDO-LID-1 for tuberculoid (T) (1.56% - 15.62%) and borderline-tuberculoid (BT) patients (7.95% - 26,13%), medium levels in the borderline-borderline (BB) (47.91% - 68.75)% and high in lepromatous (L) (93.33% - 100%) and borderline-lepromatous BL) , 88,0% - 100%). In addition, when comparing the reactional groups (RI and RII) with the non-reactive (SR) groups at the time of diagnosis, we observed a statistically significant difference between the groups, and patients with RII presented higher serological response for both anti-PGL-1 and for anti-NDO-LID-1 (66.66% - 91.66%). About the patients who developed a reaction after the initial diagnosis, they also showed significant positivity for both anti-PGL-1 and anti-NDO-LID-1 in comparison to the patients who stayed non-reactives in the study period (p < 0.0001). These results allow us to conclude that serological tests may contribute to an early diagnosis of RII, and the anti-NDO-LID-1 test demonstrated to be a better indicator.

Simultaneous analysis of multiple T helper subsets in leprosy reveals distinct patterns of Th1, Th2, Th7 and tregs markers expression in clinical forms and reactional events

Leprosy is a chronic infectious disease caused by Mycobacterium leprae. Previous studies have demonstrated that the difference among clinical forms of leprosy can be associated with the immune response of patients, mainly by T helper (Th) and regulatory T cells (Tregs). Then, aiming at clarifying the immune response, the expression of cytokines related to Th1, Th2, Th17 and Tregs profiles were evaluated by qPCR in 87 skin biopsies from leprosy patients. Additionally, cytokines and anti-PGL-1 antibodies were determined in serum by ELISA. The results showed that the expression of various targets (mRNA) related to Th1, Th2, Th17 and Tregs were significantly modulated in leprosy when compared with healthy individuals, suggesting the presence of a mixed profile. In addition, the targets related to Th1 predominated in the tuberculoid pole and side and Th2 and Tregs predominated in the lepromatous pole and side; however, Th17 targets showed a mixed profile. Concerning reactional events, Tregs markers were decreased and IL-15 was increased in reversal reaction and IL-17F, CCL20 and IL-8 in erythema nodosum leprosum, when compared with the respective non-reactional leprosy patients. Additionally, ELISA analysis demonstrated that IL-22, IL-6, IL-10 and anti-PGL-1 antibody levels were significantly higher in the serum of patients when compared with healthy individuals, and IL-10 and anti-PGL-1 antibodies were also increased in the lepromatous pole and side. Together, these results indicate that Th1, Th2 and Th17 are involved in the determination of clinical forms of leprosy and suggest that decreased Tregs activity may be involved in the pathogenesis of reactional events.

Seroreactivity to new Mycobacterium leprae protein antigens in different leprosy-endemic regions in Brazil

New Mycobacterium leprae protein antigens can contribute to improved serologic tests for leprosy diagnosis/classification and multidrug therapy (MDT) monitoring. This study describes seroreactivity to M. leprae proteins among participants from three highly endemic leprosy areas in Brazil: central-western Goiânia/Goiás (GO) (n = 225), Rondonópolis/Mato Grosso (MT) (n = 764) and northern Prata Village/Pará (PA) (n = 93). ELISA was performed to detect IgG to proteins (92f, 46f, leprosy IDRI diagnostic-1, ML0405, ML1213) and IgM to phenolic glycolipid-I (PGL-I). Multibacillary (MB) leprosy had positive rates for PGL-I that were similar to those for proteins; however, some anti-PGL-I-negative subjects were positive for proteins, suggesting that adding protein antigen to PGL-I can enhance the sensitivity of MB leprosy detection. In MT, different degrees of seroreactivity were observed and ranked for MB, former patients after MDT, paucibacillary (PB) leprosy, household contact (HHC) and endemic control (EC) groups. The seroreactivity of PB patients was low in GO and MT. HHCs from different endemic sites had similar IgG antibody responses to proteins. 46f and 92f were not recognised by most tuberculosis patients, ECs or HHCs within GO, an area with high BCG vaccination coverage. Low positivity in EC and HHC was observed in PA and MT. Our results provide evidence for the development of an improved serologic test that could be widely applicable for MB leprosy testing in Brazil.

Seroreactivity to new Mycobacterium leprae protein antigens in different leprosy-endemic regions in Brazil.

New Mycobacterium leprae protein antigens can contribute to improved serologic tests for leprosy diagnosis/classification and multidrug therapy (MDT) monitoring. This study describes seroreactivity to M. leprae proteins among participants from three highly endemic leprosy areas in Brazil: central-western Goiânia/Goiás (GO) (n = 225), Rondonópolis/Mato Grosso (MT) (n = 764) and northern Prata Village/Pará (PA) (n = 93). ELISA was performed to detect IgG to proteins (92f, 46f, leprosy IDRI diagnostic-1, ML0405, ML1213) and IgM to phenolic glycolipid-I (PGL-I). Multibacillary (MB) leprosy had positive rates for PGL-I that were similar to those for proteins; however, some anti-PGL-I-negative subjects were positive for proteins, suggesting that adding protein antigen to PGL-I can enhance the sensitivity of MB leprosy detection. In MT, different degrees of seroreactivity were observed and ranked for MB, former patients after MDT, paucibacillary (PB) leprosy, household contact (HHC) and endemic control (EC) groups. The seroreactivity of PB patients was low in GO and MT. HHCs from different endemic sites had similar IgG antibody responses to proteins. 46f and 92f were not recognised by most tuberculosis patients, ECs or HHCs within GO, an area with high BCG vaccination coverage. Low positivity in EC and HHC was observed in PA and MT. Our results provide evidence for the development of an improved serologic test that could be widely applicable for MB leprosy testing in Brazil.

Seroreactivity to new Mycobacterium leprae protein antigens in different leprosy-endemic regions in Brazil

New Mycobacterium leprae protein antigens can contribute to improved serologic tests for leprosy diagnosis/classification and multidrug therapy (MDT) monitoring. This study describes seroreactivity to M. leprae proteins among participants from three highly endemic leprosy areas in Brazil: central-western Goiânia/Goiás (GO) (n = 225), Rondonópolis/Mato Grosso (MT) (n = 764) and northern Prata Village/Pará (PA) (n = 93). ELISA was performed to detect IgG to proteins (92f, 46f, leprosy IDRI diagnostic-1, ML0405, ML1213) and IgM to phenolic glycolipid-I (PGL-I). Multibacillary (MB) leprosy had positive rates for PGL-I that were similar to those for proteins; however, some anti-PGL-I-negative subjects were positive for proteins, suggesting that adding protein antigen to PGL-I can enhance the sensitivity of MB leprosy detection. In MT, different degrees of seroreactivity were observed and ranked for MB, former patients after MDT, paucibacillary (PB) leprosy, household contact (HHC) and endemic control (EC) groups. The seroreactivity of PB patients was low in GO and MT. HHCs from different endemic sites had similar IgG antibody responses to proteins. 46f and 92f were not recognised by most tuberculosis patients, ECs or HHCs within GO, an area with high BCG vaccination coverage. Low positivity in EC and HHC was observed in PA and MT. Our results provide evidence for the development of an improved serologic test that could be widely applicable for MB leprosy testing in Brazil

The effect of iron supplementation in the diet of Dasypus novemcinctus (Linnaeus, 1758) armadillos in captivity / O efeito da suplementação com ferro na dieta de tatus Dasypus novemcinctus (Linnaeus, 1758) em cativeiro

Armadillos of the species Dasypus novemcinctus have been used as an experimental model of leprosy. Besides non-human primates, they are the only species naturally infected with Mycobacterium leprae and when experimentally inoculated, reproduce the lepromatous form of the disease producing large quantities of bacilli. This species has been maintained in captivity by numerous researchers and specific housing and feeding requirements have been developed to guarantee their survival during long experimental periods. In the "Lauro de Souza Lima" Institute, armadillos receive dog food, ground beef, boiled eggs and vitamin C. However, despite the balanced diet, anemia has been observed in some captive animals, especially in armadillos inoculated with M. leprae in advanced stages of infection. Thus, the objective of the present study was to evaluate the effect of iron sulfate supplementation in the feed provided for armadillos, both inoculated and non-inoculated with M. leprae, by means of the evaluation of their hematological profile. Fourteen armadillos received 10 mg/animal of iron sulfate (Hematofer®) diluted in sterile water mixed with their daily feed for 50 days. Hemograms and serum iron dosages for each armadillo were performed before and after supplementation. The hematocrit values increased significantly after iron supplementation, both in armadillos inoculated and non-inoculated with M. leprae. It is possible that the amount of iron in the feed is insufficient for the formation of hemoglobin, leading to microcytic anemia. Dietary supplementation with iron sulfate reversed this state, showing the importance of understanding the metabolism of exotic species for their maintenance in captivity, and thus ensuring their well-being.

O tatu da espécie Dasypus novemcinctus tem sido utilizado como modelo experimental para a hanseníase. Eles são a única espécie, além de primatas não humanos, que se apresentam naturalmente infectados pelo Mycobacterium leprae. Estes tatus, quando experimentalmente inoculados, reproduzem a forma virchoviana da hanseníase produzindo grandes quantidades de bacilos. Esta espécie tem sido mantida em cativeiro por vários pesquisadores, no entanto, alguns cuidados são necessários para garantir a sobrevivência destes por longos períodos experimentais, tais como alojamento e alimentação. No Instituto Lauro de Souza Lima, os animais em cativeiro recebem ração canina, carne bovina, ovos cozidos e vitamina C, mesmo assim, tem-se observado o desenvolvimento de quadros de anemia nestes animais, principalmente nos inoculados com o M. leprae, em estados avançados da infecção. Sendo assim, o objetivo deste estudo foi avaliar o efeito da suplementação com sulfato ferroso na alimentação fornecida aos tatus, inoculados e não inoculados com M. leprae, por meio de avaliação de seu perfil hematológico. Foram utilizados 14 animais que receberam 10 mg/animal de sulfato ferroso (Hematofer®) diluídos em água estéril e misturados diariamente à ração por 50 dias. Foram feitos hemograma e dosagem de ferro sérico de cada animal antes e após a suplementação. Os valores de hematócrito aumentaram significativamente após a suplementação, tanto nos animais inoculados como nos não inoculados com M. leprae. É possível que a quantidade de ferro na alimentação seja insuficiente para a formação da hemoglobina, levando a um quadro de anemia microcítica. A suplementação da dieta com sulfato ferroso reverteu este quadro, mostrando a importância de se conhecer o metabolismo de espécies exóticas para manutenção destas em cativeiro, assegurando seu bem estar.

Increased chitotriosidase activity in serum of leprosy patients: association with bacillary leprosy

Human phagocyte-specific chitotriosidase is associated with several diseases involving macrophage activation. Since macrophage activation plays an important role in the control of Mycobacterium leprae infection, we studied the association of chitotriosidase with leprosy both in serum and in situ in lesional skin biopsies from patients. Serum samples from 78 Indonesian leprosy patients (39 non-reactional and 39 reactional leprosy patients) and 36 healthy controls (HC) from the same endemic region were investigated. The patients were classified as multibacillary (MB, n=69) or paucibacillary (PB, n=9) based on the bacterial index in slit-skin smears. Thirty-six of the reactional patients had erythema nodosum leprosum (ENL), while only 3 had reversal reaction (RR). Follow-up serum samples after corticosteroid treatment were also obtained from 17 patients with ENL and one with RR. Multibacillary (MB) patients showed increased chitotriosidase activity in serum as compared to paucibacillary (PB) patients and healthy controls. Although no significant difference was observed between reactional and the corresponding non-reactional groups, ENL showed significantly higher chitotriosidase activity as compared to HC. Furthermore, corticosteroid treatment resulted in significant decline of enzyme activity in ENL sera. Chitotriosidase activity correlated with levels of neopterin, another macrophage activation marker, but not with IL-6, IFN-gamma, TNF-alpha and IL-10. Immunohistochemical staining of 6 MB (LL=5, BL=1) lesional skin sections from stored material showed positive staining for chitotriosidase within lipid-laden macrophages suggesting that macrophages are the source of the enzyme detected in serum. Thus, serum chitotriosidase activity is potentially useful in distinguishing MB from PB leprosy and in monitoring response to therapy in ENL.

A reação de mitsuda em tatus Dasypus novemcinctus utilizando antigeno humano e antigeno derivado de tatus / Mitsuda reaction in armadillos Dasypus novemcinctus using human and armadillo derived antigens

O tatu foi um modelo experimental importante para o estudo da hanseniase, alem de ser ainda uma importante fonte para coleta de bacilos. Apesar dos inumeros relatos de estudos em tatus de especie Dasypus novemcinctus, pouco se sabe sobre a real susceptibilidade desta especie ao bacilo de Hansen apos inoculaçao experimental com M. leprae. Alguns autores relatam que cerca de 80 por cento dos animais desenvolveriam a doença quando infectados. No Brasil, a inoculaçao experimental desta especie resultou em inoculaçoes positiva em apenas dois momento, tendo ate sido levantada a hipotese de estes animais serem mais resistentes a infecçao experimental. No presente estudo, utilizou-se a resposta ao nantigeno de Mitsuda como um indicador de resposta imune celular de tatus ao M. leprae...

Mitsuda reaction in armadillos Dasypus novemcinctus using human and armadillo derived antigens

The armadillo has been an important experimental model for leprosy, besides it is still an important resource for bacilli. Despite the innumerous studies about armadillos of the Genus Dasypus, little is known about the real susceptibility of this species to the Hansen’s bacillus after experimental infection with M. leprae. Many authors have reported that 80 per cent of the inoculated animal will develop the disease. In Brazil, positive inoculation of this species was obtained only twice, being raised the hypothesis that these animals are more resistant to experimental infection. In the present study the response to the Mitsuda antigen was used as an indicative of cellular immune response to M. leprae in armadillos. Twenty one animals were tested with two Mitsuda antigen preparations, human derived (4,2 x 109 bacilli/ml) and armadillo derived (1,6 x 108 bacilli/ml) antigens. Response after 28 days of intradermal testing showed that most of the animals presented an infiltrate composed by grouped macrophages with vacuolated cytoplasm and rare lymphocytes. This response resembles lepromatous leprosy in humans and suggests that these animals would be susceptible to development of disseminated leprosy when successfully inoculated. Bacilloscopy in these animals varied from 3+ to 4+ according to Ridley’s scale (1966). Two animals developed a granulomatous reaction with borderline pattern and bacilloscopy varying from 1+ to 3+.

Quantificação dos niveis sericos de anticorpos anti-PGL-I, neopterina e proteina C reativa em pacientes com hanseniase durante a poliquimioterapia / ?

A hanseniase e doença infecciosa, causada pelo Mycobacterium leprae, um parasita intracelular obrigatorio nao cultivavel em meios artificiais. Esta doença pode se manifestar sob amplo espectro clinico, correspondendo a distintos padroes da resposta imunologica do hospedeiro ao M. leprae. Em um polo deste espectro, esta a forma de resistencia ao bacilo, a hanseniase tuberculoide (HT), na qual se desenvolve acentuada resposta imune celular especifica com efetivo controle da mutilaçao bacilar. O outro polo do espectro esta representado pela hanseniase virchoviana (HV), forma de baixa resistencia, em que a resposta imune celular seletivamente falha em eliminar o bacilo do organismo, resultando na disseminaçao da doença. O grupo dimorfo (HD) apresenta manifestaçoes intermediarias variaveis entre HT e HV, de acordo com o grau de resposta imune ao M. leprae. Considerando que na hanseniase existem poucos estudos avaliando os niveis sericos de anticorpos anti-PGL-I, neopterina e proteina C reativa (CRP) no momento do diagnostico e durante o tratamento poliquimioterapico, realizamos este estudo com os sguintes objetivos: A. Avaliar a resposta imune e inflamatoria de pacientes com hanseniase no momento do diagnostico e aos 2, 4, 6 e 12 meses de tratamento com poliquimioterapia (PQT) e nos estados reacionais, mediante a determinaçao dos niveis sericos de anti-PGL-I, de neopterina e de CRP....

Factors influencing the development of leprosy: an overview

The clinical manifestations of leprosy vary, seemingly depending on the host's immune response. Mode and route of infection, such as skin versus nasal mucosa, insect bites, sexual and gastroenteral transmission, together with genetic factors that may contribute to the outcome of the infection, including HLA, Lewis factor, Nramp1 and more subtle inherited alterations, are discussed. It is theorized that a balance between host responses elicited by different routes of infection and size and spacing of inocula is responsible for the clinical and immunological manifestations of the disease. Genetic factors and contact with environmental microorganisms may modulate these responses. The final result, resistance, delayed-type hypersensitivity, tolerance, disease or no disease, spectrum and reactions, is most likely reached via the orchestration of the induced cyto- and chemokines