CCL2 and IFN-γ serum levels as biomarkers for subclinical infection in household contacts of leprosy patients.

Leprosy, also known as Hansen's disease, is a long-term infection by the bacteria Mycobacterium leprae, and actually still persists as a serious public health problem. The clinical parameters are used for diagnosis, however, some studies have indicated the selection of a set of biomarkers of subclinical infection, both serological and cellular, that allow the early diagnosis. Some cytokines and chemokines have been differentially expressed in index cases (paucibacillary and multibacillary patients) and household contacts (HHC), and may present a potential biomarker of M. leprae subclinical infection. Thus, the aim of this study was to analyze the variations in the profile of cytokines and chemokines, longitudinally, between index cases and their household contacts with a view to identifying possible biomarkers with differential expression, which may guide the early subclinical infection in household contacts. A longitudinal study was carried out between 2014 and 2015. The serum levels of the cytokines and chemokines were measured in all patient samples by CBA (Cytometric Bead Array). We observed a reduction of IL-4 and IL-17 expression of HHC group in the second evaluation (T1), as also a reduction of IL-17 in MB. We observed increased expression of IL-2 in PB patients as well. HHC, PB and MB showed a similar reduction profile of the chemokines CXCL8, CXCL9 and CXCL10 from T0 to T1. Interestingly, only serological levels of CCL2 are increased after a follow-up of HHC group, and this group, but not PB and MB patients, showed a significant association and a negative correlation between CCL2 and IFN-γ. The present study showed for the first time a similarity in the immunological scenario between HHC, PB and MB patients. In addition, this work highlights CCL2 chemokine in association with IFN-γ as possible biomarkers of subclinical infection of HHC, as also a parameter of early infection monitoring.

Immunological biomarkers of subclinical infection in household contacts of leprosy patients.

Hansen's disease (or leprosy) still persists as a serious public health issue. Its diagnosis is based primarily on the detection of clinical signs that are characteristic of the disease. Studies have pointed to the selection of a set of serological and cellular biomarkers of subclinical infection that result in an efficient diagnosis. The aim of this study was compare index cases and their household contacts to identify differentially expressed biomarkers of immune response in leprosy that could provide reliable evidence of subclinical infection in household contacts. The study population consisted of index cases with multibacillary form (IC, n = 13) and their household contacts (HC, n = 14). Serum cytokines and chemokines were quantified using the cytometric beads array (CBA) system. The humoral response was assessed by ELISA test. Flow cytometry was used to characterize the cellular immune response. Monocyte and CD4 + T lymphocytes frequency was significantly higher in IC. Both CD4+ and CD8 + T lymphocytes had a reduced CD25 expression in HC. The immunoglobulin (Ig)M profile anti- NDO-HSA, LID-1, and NDOLID antigens was significantly higher in IC. This study points to the monocyte and CD4+ lymphocyte frequency, as well as specific IgM profile, as predictors of subclinical infection in the household contacts.

Regulatory T cells: Friends or foe in human Mycobacterium leprae infection?

Regulatory T cells (Tregs) are known to control immune responses by suppressing the antigen-presenting and effector T cells. Some mechanisms adopted by Tregs in combating Mycobacterium infections have been proposed. Nevertheless, in M. leprae infection, also known as leprosy or Hansen's disease, the role of Tregs has not been completely elucidated. Using multicolor flow cytometry, we evaluated the expression of different cell surface and intracellular molecules present in Tregs from peripheral blood samples of leprosy patients. Before initiating treatment, thirteen new cases of leprosy were grouped according to the Ridley-Jopling classification in to the paucibacilary (PB) or multibacilary (MB) group. Fifteen non-infected individuals (NI) were included as control subjects. Tregs were higher in the MB group than in the NI group. Tregs also co-expressed high amounts of PD1 and PDL-1, indicating that these cells could induce apoptosis of effector cells and simultaneously prevent their own apoptosis. Our data showed that compared to the NI group, Tregs from the PB group expressed higher levels of CD95L, which may be associated with other apoptotic pathways that may decrease Tregs in these patients. Correlation analysis reinforced that PD1 and CD95L are efficient apoptosis' pathway that decreased levels of Tregs in the NI and PB groups. We also observed significant differences in cytokine expression of Tregs from the PB and MB groups. Compared to the NI group, Tregs from the MB group showed higher IL-17 expression; however, compared to the PB group, the expression of IL-10 in Tregs from the MB group was lower, suggesting inefficient control of inflammation. Therefore, we concluded that different pathways were involved in Treg-induced suppression of leprosy. Moreover, Treg-mediated regulation of inflammation via IL-10 and IL-17 expression in leprosy patients was inefficient. Thus, we propose that during M. leprae infection, Tregs may impair the immune responses elicited against this bacillus, favor bacterial replication, and aid in persistence of a disseminated multibacillary disease.

Specific antigen serologic tests in leprosy: implications for epidemiological surveillance of leprosy cases and household contacts.

BACKGROUND: There is a lack of straightforward tests for field application and known biomarkers for predicting leprosy progression in infected individuals. OBJECTIVE: The aim was to analyse the response to infection by Mycobacterium leprae based on the reactivity of specific antigens: natural disaccharide linked to human serum albumin via an octyl (NDOHSA), a semisynthetic phenolic glycolipid-I (PGL-I); Leprosy Infectious Disease Research Institute Diagnostic-1 (LID-1) and natural disaccharide octyl - Leprosy Infectious Disease Research Institute Diagnostic-1 (NDOLID). METHODS: The study population consisted of 130 leprosy cases diagnosed between 2010 and 2015 and 277 household contacts. An enzyme-linked immunosorbent assay (ELISA) was used to analyse the reactivity of antibodies against NDOHSA, LID-1 and NDOLID. The samples and controls were tested in duplicate, and the antibody titer was expressed as an ELISA index. Data collection was made by home visits with application of questionnaire and dermatological evaluation of all household contacts to identify signs and symptoms of leprosy. FINDINGS: Significant differences in the median ELISA results were observed among leprosy cases in treatment, leprosy cases that had completed treatment and household contacts. Higher proportions of seropositivity were observed in leprosy cases in treatment. Seropositivity was also higher in multibacillary in relation to paucibacillary, with the difference reaching statistical significance. Lower titers were observed among cases with a longer treatment time or discharge. For household contacts, the differences according to the clinical characteristics of the leprosy index case were less pronounced than expected. Other factors, such as the endemicity of leprosy, exposure outside the residence and genetic characteristics, appeared to have a greater influence on the seropositivity. MAIN CONCLUSIONS: Serologic tests could be used as auxiliary tools for determining the operational classification, in addition to identifying infected individuals and as a strategy for surveillance of household contacts.

Specific antigen serologic tests in leprosy: implications for epidemiological surveillance of leprosy cases and household contacts

BACKGROUND There is a lack of straightforward tests for field application and known biomarkers for predicting leprosy progression in infected individuals. OBJECTIVE The aim was to analyse the response to infection by Mycobacterium leprae based on the reactivity of specific antigens: natural disaccharide linked to human serum albumin via an octyl (NDOHSA), a semisynthetic phenolic glycolipid-I (PGL-I); Leprosy Infectious Disease Research Institute Diagnostic-1 (LID-1) and natural disaccharide octyl - Leprosy Infectious Disease Research Institute Diagnostic-1 (NDOLID). METHODS The study population consisted of 130 leprosy cases diagnosed between 2010 and 2015 and 277 household contacts. An enzyme-linked immunosorbent assay (ELISA) was used to analyse the reactivity of antibodies against NDOHSA, LID-1 and NDOLID. The samples and controls were tested in duplicate, and the antibody titer was expressed as an ELISA index. Data collection was made by home visits with application of questionnaire and dermatological evaluation of all household contacts to identify signs and symptoms of leprosy. FINDINGS Significant differences in the median ELISA results were observed among leprosy cases in treatment, leprosy cases that had completed treatment and household contacts. Higher proportions of seropositivity were observed in leprosy cases in treatment. Seropositivity was also higher in multibacillary in relation to paucibacillary, with the difference reaching statistical significance. Lower titers were observed among cases with a longer treatment time or discharge. For household contacts, the differences according to the clinical characteristics of the leprosy index case were less pronounced than expected. Other factors, such as the endemicity of leprosy, exposure outside the residence and genetic characteristics, appeared to have a greater influence on the seropositivity. MAIN CONCLUSIONS Serologic tests could be used as auxiliary tools for determining the operational classification, in addition to identifying infected individuals and as a strategy for surveillance of household contacts.

Cytokines as biomarkers to monitoring the impact of multidrug therapy in immune response of leprosy patients.

Leprosy or Hansen's disease is a chronic infectious disease of the skin and nerves, caused by the intracellular bacilli Mycobacterium leprae. It is characterized by a spectrum of clinical forms depending on the host's immune response to M. leprae. Patients with tuberculoid (TT) leprosy have strong cell-mediated immunity (CMI) with elimination of the bacilli, whereas patients with lepromatous (LL) leprosy exhibit defective CMI to M. leprae. Despite advances in the understanding of the pathogenesis of leprosy and the development of new therapeutic strategies, there is a need for the identification of biomarkers which be used for early diagnosis and to discrimination between different forms of the disease, as prognostic markers. Here, we analyzed the serum levels of IL-1ß, IL-6, IL-8, IL-10, IL-12p70, IL-13, IL-17A, IFN-γ and TNF in order to address the contribution of these cytokines in late phase of M. leprae infection, and the impact of multidrug therapy (MDT). Our results demonstrated that patients of LL group presented higher expression of serum levels of inflammatory cytokines before MDT, while TT patients presented a balance between inflammatory and regulatory cytokines. MDT changes the profile of serum cytokines in M. leprae infected patients, as evidenced by the cytokine network, especially in TT patients. LL patients displayed a multifaceted cytokine system characterized by strong connecting axes involving inflammatory/regulatory molecules, while TT patients showed low involvement of regulatory cytokines in network overall. Cytokines can be identified as good biomarkers of the impact of MDT on the immune system and the effectiveness of treatment.

Integrative literature review of the reported uses of serological tests in leprosy management.

An integrative literature review was conducted to synthesize available publications regarding the potential use of serological tests in leprosy programs. We searched the databases Literatura Latino-Americana e do Caribe em Ciências da Saúde, Índice Bibliográfico Espanhol em Ciências da Saúde, Acervo da Biblioteca da Organização Pan-Americana da Saúde, Medical Literature Analysis and Retrieval System Online, Hanseníase, National Library of Medicine, Scopus, Ovid, Cinahl, and Web of Science for articles investigating the use of serological tests for antibodies against phenolic glycolipid-I (PGL-I), ML0405, ML2331, leprosy IDRI diagnostic-1 (LID-1), and natural disaccharide octyl-leprosy IDRI diagnostic-1 (NDO-LID). From an initial pool of 3.514 articles, 40 full-length articles fulfilled our inclusion criteria. Based on these papers, we concluded that these antibodies can be used to assist in diagnosing leprosy, detecting neuritis, monitoring therapeutic efficacy, and monitoring household contacts or at-risk populations in leprosy-endemic areas. Thus, available data suggest that serological tests could contribute substantially to leprosy management.

Integrative literature review of the reported uses of serological tests in leprosy management

Abstract: An integrative literature review was conducted to synthesize available publications regarding the potential use of serological tests in leprosy programs. We searched the databases Literatura Latino-Americana e do Caribe em Ciências da Saúde, Índice Bibliográfico Espanhol em Ciências da Saúde, Acervo da Biblioteca da Organização Pan-Americana da Saúde, Medical Literature Analysis and Retrieval System Online, Hanseníase, National Library of Medicine, Scopus, Ovid, Cinahl, and Web of Science for articles investigating the use of serological tests for antibodies against phenolic glycolipid-I (PGL-I), ML0405, ML2331, leprosy IDRI diagnostic-1 (LID-1), and natural disaccharide octyl-leprosy IDRI diagnostic-1 (NDO-LID). From an initial pool of 3.514 articles, 40 full-length articles fulfilled our inclusion criteria. Based on these papers, we concluded that these antibodies can be used to assist in diagnosing leprosy, detecting neuritis, monitoring therapeutic efficacy, and monitoring household contacts or at-risk populations in leprosy-endemic areas. Thus, available data suggest that serological tests could contribute substantially to leprosy management.

Factors associated with anti-phenolic glycolipid-I seropositivity among the household contacts of leprosy cases.

BACKGROUND: The diagnosis of leprosy is based on clinical symptoms of the disease, which may not be sufficient to ensure early diagnosis. The development of effective tools for the early detection of infection, such as rapid tests that can be applied by non-specialists for early-stage leprosy identification, has been considered a research priority and may contribute to overcoming the complications associated with late diagnosis. The aim of this study was to analyze the factors associated with anti-phenolic glycolipid-I (PGL-I) seropositivity among the household contacts of leprosy cases. METHODS: A cross-sectional study of individuals from the northeastern municipalities of the state of Minas Gerais, Brazil, was performed. Anti-PGL-I seropositivity was evaluated by assessing specific antibody production using the ML Flow test. A Poisson regression with a robust error variance was used to evaluate the relationship between anti-PGL-I seropositivity and the independent variables investigated. RESULTS: The overall anti-PGL-I seropositivity was 13.5 %, and among the contacts of leprosy cases that were classified as paucibacillary or multibacillary, it was 8.4 and 17.3 %, respectively. The factors associated with the variation of anti-PGL-I seropositivity among the study population were the presence of signs suggestive of leprosy (PR = 3.68; 95 % CI: 1.56-8.71), the operational leprosy classification (PR = 2.17; 95 % CI: 1.22-3.86) and grade 1 (PR = 1.83; 95 % CI: 1.02-3.26) or grade 2 disability (PR = 2.42; 95 % CI: 1.02-5.47) of the index leprosy case. CONCLUSIONS: The presence of signs suggestive of leprosy and the operational classification of leprosy cases were associated with anti-PGL-I seropositivity. The serological tests available for leprosy are not considered to be diagnostic tests but can be used as auxiliary assessments in combination with clinical parameters to identify exposed individuals at high risk of developing leprosy and those exhibiting the initial stages of this disease.

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.