In Situ versus Systemic Immune Response in the Pathogenesis of Cutaneous Leishmaniasis

The role of the immune response in the pathogenesis of cutaneous leishmaniasis (CL) due to Leishmania (Viannia) braziliensis is predominantly carried out via blood cells. Here, we evaluate whether cytokine production by peripheral blood mononuclear cells (PBMCs) reflects what has been documented at the lesion site. The participants included 22 CL patients diagnosed with a positive PCR. PBMCs were stimulated for 72 h with a soluble leishmania antigen (SLA). Biopsies obtained from the edge of the ulcers were incubated for the same period. Cytokines in supernatants were assessed via ELISA. TNF, IL-1β, IL-6, IL-17, and granzyme B (GzmB) were higher in the supernatants of biopsies than in PBMCs, but IFN-γ was higher in the supernatants of PBMCs than in biopsies. There was a positive correlation between IFN-γ and TNF in PBMCs, and an inverse correlation between TNF and IL-10 in the cells from the lesion site. A strong correlation between IL-1β, IL-17, and GzmB was observed in the biopsies, and a positive correlation was detected between these cytokines and the lesion size. Our results indicate that the immune response in L. braziliensis lesions is different from that observed in peripheral blood, and our data suggest that in addition to IL-1β and GzmB, IL-17 participates in the pathology of CL

Early Cutaneous Leishmaniasis Patients Infected With Leishmania braziliensis Express Increased Inflammatory Responses After Antimony Therapy

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2018-02-28T17:29:50Z No. of bitstreams: 1 Costa RS Early Cutaneous Leishmaniasis Patients Infected....pdf: 1578790 bytes, checksum: ebe3b7a6ad798d68d377b1aafe5d1de2 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2018-02-28T17:51:42Z (GMT) No. of bitstreams: 1 Costa RS Early Cutaneous Leishmaniasis Patients Infected....pdf: 1578790 bytes, checksum: ebe3b7a6ad798d68d377b1aafe5d1de2 (MD5)Made available in DSpace on 2018-02-28T17:51:42Z (GMT). No. of bitstreams: 1 Costa RS Early Cutaneous Leishmaniasis Patients Infected....pdf: 1578790 bytes, checksum: ebe3b7a6ad798d68d377b1aafe5d1de2 (MD5) Previous issue date: 2018National Institutes of Health (Grant U01 AI088650-06)Universidade Federal da Bahia. Hospitalar Universitário Professor Edgard Santos. Serviço de Imunologia. Salvador, BA, BrasilUniversidade Federal da Bahia. Hospitalar Universitário Professor Edgard Santos. Serviço de Imunologia. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Instituto Nacional de Ciência e Tecnologia em Doenças Tropicais. Salvador, BA, BrasilUniversidade Federal da Bahia. Hospitalar Universitário Professor Edgard Santos. Serviço de Imunologia. Salvador, BA, BrasilUniversidade Federal da Bahia. Hospitalar Universitário Professor Edgard Santos. Serviço de Imunologia. Salvador, BA, BrasilUniversidade Federal da Bahia. Hospitalar Universitário Professor Edgard Santos. Serviço de Imunologia. Salvador, BA, BrasilUniversidade Federal da Bahia. Hospitalar Universitário Professor Edgard Santos. Serviço de Imunologia. Salvador, BA, Brasil / University of Pennsylvania. School of Veterinary Medicine. Department Pathobiology. Philadelphia, PAUniversidade Federal da Bahia. Hospitalar Universitário Professor Edgard Santos. Serviço de Imunologia. Salvador, BA, BrasilUniversidade Federal da Bahia. Hospitalar Universitário Professor Edgard Santos. Serviço de Imunologia. Salvador, BA, BrasilUniversidade Federal da Bahia. Hospitalar Universitário Professor Edgard Santos. Serviço de Imunologia. Salvador, BA, BrasilUniversidade Federal da Bahia. Hospitalar Universitário Professor Edgard Santos. Serviço de Imunologia. Salvador, BA, Brasil / Universidade Federal da Bahia. Instituto Nacional de Ciência e Tecnologia em Doenças Tropicais. Salvador, BA, BrasilUniversity of Pennsylvania. School of Veterinary Medicine. Department Pathobiology. Philadelphia, PAUniversidade Federal da Bahia. Hospitalar Universitário Professor Edgard Santos. Serviço de Imunologia. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Instituto Nacional de Ciência e Tecnologia em Doenças Tropicais. Salvador, BA, BrasilEarly cutaneous leishmaniasis (ECL) is characterized by a nonulcerated papular lesion and illness duration less than 30 days. Approximately 4 weeks later, the cutaneous leishmaniasis (CL) ulcers appear. We were surprised to find that failure after antimony therapy (Sb5) is higher in ECL than CL. We hypothesize that the inflammatory response in ECL patients may increase during Sb5 therapy, which leads to treatment failure. Methods. A cohort of 44 ECL patients infected by Leishmania braziliensis was established to evaluate the response to Sb5 and to compare immunologic responses in ECL patients with CL and healthy subjects. Results. A hierarchical clustering based on cytokine levels showed a weak positive correlation between proinflammatory cytokine levels and those patients that failed Sb5 treatment. Although Sb5 therapy decreased interferon-γ and tumor necrosis factor levels in CL patients, we were surprised to find that an increase in these cytokines was observed in ECL patients. Moreover, interleukin (IL)-10 was less able to down-modulate immune responses in ECL. Conclusions. The enhanced production of proinflammatory cytokines, due in part to the decreased ability of IL-10 to down-modulate immune response during therapy in ECL, promotes the development and persistence of leishmania ulcer despite antimony therapy

Lesions of CL patients produce MMP-9.

<p><i>A</i>, Heatmap showing expression of MMPs and TIMP-1 from microarray profile. Biopsies from CL patients (<i>n</i> = 26) and normal skin (<i>n</i> = 10) were obtained and unbiased microarray was performed on biopsies mRNA. Average fold change (FC) for each gene in lesion samples relative to normal skin controls is shown. <i>B</i>, Ratio between MMP-9 and TIMP-1 genes expression. Biopsies from CL patients (<i>n</i> = 26) and normal skin (<i>n</i> = 10) were obtained and unbiased microarray was performed on biopsies mRNA. <i>C</i>, MMP-9 levels in biopsies culture supernatants from CL patients (<i>n = 6</i>) and healthy subjects (HS) (<i>n = 5</i>), determined by ELISA after the biopsies been cultured for 12 h in absence of stimuli. **p<0.005.</p

TNF enhances MMP-9 production in CL patients.

<p><i>A</i>, PBMC from healthy subjects (HS) <i>(n = 12)</i> and CL patients (<i>n = 19)</i> were cultured in the presence or absence of SLA (5 µg/ml) for 72 hours, and TNF concentrations were determined by ELISA on supernatants. <i>B</i>, PBMC from HS (<i>n = 5</i>) were cultured in the presence or absence of recombinant TNF (5 ng/ml) for 24 hours and the levels of MMP-9 were determined by ELISA. <i>C</i>, PBMC from HS <i>(n = 6)</i> and CL patients <i>(n = 6)</i> was cultured in presence or absence of anti-TNF antibodies (10 µg/ml) for 24 hours. The levels of MMP-9 were determined by ELISA on supernatants. *p<0.05; ***p<0.0005.</p