ABSTRACT
Diagnosis of leprosy is a major obstacle to disease control and has been compromised in the past due to the lack of specific reagents. We have used comparative genome analysis to identify genes that are specific to Mycobacterium leprae and tested both recombinant proteins and synthetic peptides from a subset of these for immunological reactivity. Four unique recombinant proteins (ML0008, ML0126, ML1057, and ML2567) and a panel of 58 peptides (15 and 9 mer) were tested for IFN-gamma responses in PBMC from leprosy patients and contacts, tuberculosis patients, and endemic and nonendemic controls. The responses to the four recombinant proteins gave higher levels of IFN-gamma production, but less specificity, than the peptides. Thirty-five peptides showed IFN-gamma responses only in the paucibacillary leprosy and household contact groups, with no responses in the tuberculosis or endemic control groups. High frequencies of IFN-gamma-producing CD4+ and CD8+ T cells specific for the 15- and 9-mer peptides were observed in the blood of a paucibacillary leprosy patient. 9-mer peptides preferentially activated CD8+ T cells, while the 15-mer peptides were efficient in inducing responses in both the CD4+ and CD8+ T cell subsets. Four of the six 9-mer peptides tested showed promising specificity, indicating that CD8+ T cell epitopes may also have diagnostic potential. Those peptides that provide specific responses in leprosy patients from an endemic setting could potentially be developed into a rapid diagnostic test for the early detection of M. leprae infection and epidemiological surveys of the incidence of leprosy, of which little is known.
Subject(s)
Antigens, Bacterial , Leprosy/diagnosis , Mycobacterium leprae/immunology , Antigens, Bacterial/immunology , CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Case-Control Studies , Cells, Cultured , Diagnostic Techniques and Procedures/standards , Genome, Bacterial , Humans , Interferon-gamma/analysis , Interferon-gamma/biosynthesis , Leprosy/immunology , Leprosy/microbiology , Leukocytes, Mononuclear/immunologyABSTRACT
Tumor necrosis factor alpha (TNFalpha) plays a key role in orchestrating the complex events involved in inflammation and immune response. The presence of single nucleotide polymorphisms (SNPs) within the promoter region of the TNFa gene has been associated with a number of diseases. The aim of this study was to investigate the distribution of polymorphisms at positions -238 (G/A) and -308 (G/A) at the TNFalpha promoter, and its association to the outcome of different clinical forms of leprosy. Furthermore, the bacteriological index (BI) was evaluated among genotyped multibacillary (MB) patients in order to investigate the possible influence of each polymorphism on the bacterial load. This study included a total of 631 leprosy patients being 401 MB and 230 paucibacillary (PB), that was further separated according to its ethnicity (Afro- and Euro-Brazilians). The combination of SNPs in haplotypes generated three different arrangements: TNFG-G, TNFG-A and TNFA-G. In spite of the marked differences observed in the frequency of the haplotypes along the ethnic groups, no statistical differences were observed in haplotype frequencies between MB and PB patients. The BI analyses showed a lower bacteriological index among the -308 carriers, while the BI of the -238 carriers was higher. Although no significance has been achieved in this analysis regarding the influence of the polymorphisms to the development of the clinical outcome, it seems that in a different stage (among the MB patients) the polymorphisms could contribute to the degree of severity observed.
Subject(s)
Leprosy/genetics , Mycobacterium leprae/growth & development , Polymorphism, Single Nucleotide , Tumor Necrosis Factor-alpha/genetics , Black People , Brazil , DNA/chemistry , DNA/genetics , Female , Haplotypes , Humans , Leprosy/immunology , Leprosy/microbiology , Male , Promoter Regions, Genetic , Regression Analysis , Tumor Necrosis Factor-alpha/immunology , White PeopleSubject(s)
Immunization, Passive , Immunotherapy , Immunotherapy, Active , Neoplasms , Biological Therapy/methods , Adoptive Transfer , Immunotoxins , Lymphokines , VaccinesABSTRACT
Dados recentes sugerem que o curso clinico dos estados reacionais na lepra esta estritamente relacionado a liberacao local ou sistemica de citocinas. Neste estudo, pacientes com ENL (erythema nodosum leprosum) foram agrupadas segundo a intensidade de seus sintomas clinicos. Os aspectos clinicos e imunologicos do ENL e os efeitos desses parametros na carga bacilar foram estabelecidos em conjuncao com a resposta imune in vitro desses pacientes, a antigenos microbacterianos. Em 10 de 17 pacientes testados, o indice bacteriano (IB) foi reduzido em pelo menos 1 log desde o diagnostico da lepra ate o aparecimento do primeiro episodio reacional, comparado a uma reducao esperada de 0.3 log no grupo reacional no mesmo periodo de MDT (multidrogaterapia)...
