title: Identification of specific proteins and peptides in Mycobacterium leprae suitable for the selective diagnosis of leprosy.
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.
authors.affiliation: Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO 80523, USA. John.Spencer@colostate.edu
authors.forename: John S
authors.initials: JS
authors.lastname: Spencer
authors.forename: Hazel M
authors.initials: HM
authors.lastname: Dockrell
authors.forename: Hee Jin
authors.initials: HJ
authors.lastname: Kim
authors.forename: Maria A M
authors.initials: MA
authors.lastname: Marques
authors.forename: Diana L
authors.initials: DL
authors.lastname: Williams
authors.forename: Marcia V S B
authors.initials: MV
authors.lastname: Martins
authors.forename: Marcio L F
authors.initials: ML
authors.lastname: Martins
authors.forename: Monica C B S
authors.initials: MC
authors.lastname: Lima
authors.forename: Euzenir N
authors.initials: EN
authors.lastname: Sarno
authors.forename: Geraldo M B
authors.initials: GM
authors.lastname: Pereira
authors.forename: Haroldo
authors.initials: H
authors.lastname: Matos
authors.forename: Leila S
authors.initials: LS
authors.lastname: Fonseca
authors.forename: Elisabeth P
authors.initials: EP
authors.lastname: Sampaio
authors.forename: Thomas H M
authors.initials: TH
authors.lastname: Ottenhoff
authors.forename: Annemieke
authors.initials: A
authors.lastname: Geluk
authors.forename: Sang-Nae
authors.initials: SN
authors.lastname: Cho
authors.forename: Neil G
authors.initials: NG
authors.lastname: Stoker
authors.forename: Stewart T
authors.initials: ST
authors.lastname: Cole
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Brennan
authors.forename: Maria C V
authors.initials: MC
authors.lastname: Pessolani
mesh_terms: D000942:Antigens, Bacterial / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D016022:Case-Control Studies; D002478:Cells, Cultured; D019937:Diagnostic Techniques and Procedures / Q000592:standards; D016680:Genome, Bacterial; D006801:Humans; D007371:Interferon-gamma / Q000032:analysis / Q000096:biosynthesis; D007918:Leprosy / Q000175:diagnosis* / Q000276:immunology / Q000382:microbiology; D007963:Leukocytes, Mononuclear / Q000276:immunology; D009166:Mycobacterium leprae / Q000276:immunology*
chemical_list: D000942:Antigens, Bacterial; D007371:Interferon-gamma