Assuntos
Adalimumab/efeitos adversos , Antirreumáticos/efeitos adversos , Artrite Psoriásica/complicações , Leishmaniose Cutânea/etiologia , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Uveíte Anterior/complicações , Adalimumab/uso terapêutico , Idoso , Antirreumáticos/uso terapêutico , Artrite Psoriásica/tratamento farmacológico , Artrite Psoriásica/fisiopatologia , Suscetibilidade a Doenças , Quimioterapia Combinada , Antígeno HLA-B27/análise , Humanos , Imunossupressores/uso terapêutico , Leishmaniose Cutânea/fisiopatologia , Masculino , Metotrexato/uso terapêutico , Prednisona/uso terapêutico , Fator de Necrose Tumoral alfa/fisiologia , Uveíte Anterior/tratamento farmacológico , Uveíte Anterior/fisiopatologiaRESUMO
Este trabajo utilizó un modelo macrófago humano-amastigote como herramienta para recrear in vitro la infección causada por aislados de pacientes con fracaso terapéutico y valorar su utilidad en la identificación de aislados de Leishmania con fenotipo quimio-resistente. Objetivos: (1) Evaluar un modelo in vitro de macrófago humano-amastigote y (2) Determinar su utilidad en la identificación de aislados de Leishmania con fenotipo quimio-resistente. Métodos: Se evaluó un protocolo de purificación basado en la capacidad de los monocitos de adherirse al plástico. Monocitos purificados de sangre humana fueron infectados con promastigotes metacíclicos de especies de referencia y aislados de Leishmania de tres pacientes con falla terapéutica a antimoniales. Se determinó el porcentaje de infección inicial y el efecto leishmanicida de glucantime, anfotericinaB y pentamidina; se correlacionó la capacidad leishmanicida con los niveles de producción de óxido nítrico en cada condición estudiada. Resultados: Los resultados sugieren que el modelo macrófago humano-amastigote empleado recrea in vitro la infección causada por especies de referencia, o con aislados de pacientes con fracaso terapéutico. Adicionalmente sugieren que en monocitos infectados (1) con el aislado VE98MR no puede definirse una IC50 para glucantime ni para pentamidina y (2) con el aislado VE96ZC no puede definirse una IC50 para glucantime mas si para pentamidina. De igual forma, se evidencia una disminución efectiva del porcentaje de infección susceptible a anfotericina-B, para todos los aislados y cepas de referencia. El efecto leishmanicida no se correlaciona con aumentos significativos de la producción de óxido nítrico. Conclusiones: El modelo macrófago humano-amastigote empleado constituye una prueba de concepto que permitió identificar como aislados potencialmente quimio-resistentes a L. (L.) amazonensis (VE98MR) y L. (L.) mexicana (VE96ZC), mas no al aislado L. (L.) amazonensis (VE2000MM)(AU)
This work used a human-amastigote macrophage model as a tool to recreate in vitro infection caused by isolates from patient's with therapeutic failure and assess its usefulness in the identification of chemo-resistant Leishmania isolates. Objectives: (1) Evaluate in vitro a human-amastigote macrophage model and (2) determine its usefulness in the identification of Leishmania isolates with chemo-resistant phenotype. Methods: A purification protocol based on the ability of monocytes to adhere to plastic was evaluated. Monocytes purified from human blood were infected with metacyclic promastigotes of reference species and Leishmania isolates from three patients with antimonial therapeutic failure. The percentage of initial infection and the leishmanicidal effect of glucantime, amphotericin-B and pentamidine were determined; the leishmanicidal capacity was correlated with the levels of nitric oxide production in each condition studied. Results: Results suggest that the human-amastigote macrophage model recreates in vitro the infection caused by reference species, or isolates from patients with therapeutic failure. In addition, they suggest that (1) an effective IC50 for glucantime and pentamidine could not be defined in monocytes infected with the isolate VE98MR and (2) an effective IC50 for pentamidine but nor for glucantime could be defined in monocytes infected with the isolate VE96ZC. On the contrary, an effective decrease in the percentage of infection susceptible to amphotericin-B was observed for all isolates and reference strains. The leishmanicidal effect did not correlate with significant increases in nitric oxide production. Conclusion: The human-amastigote macrophage model used constitutes a proof of concept to identify as potentially chemo-resistant isolates L. (L.) amazonensis (VE98MR) and L. (L.) mexicana (VE96ZC), but not L (L.) amazonensis (VE2000MM)(AU)
Assuntos
Humanos , Masculino , Feminino , Técnicas In Vitro/métodos , Leishmaniose Cutânea/fisiopatologia , Leishmaniose Cutânea/epidemiologia , Leishmaniose Cutânea/virologia , Medicina Tropical , Saúde Pública , Tratamento Farmacológico , Ativação de MacrófagosRESUMO
BACKGROUND: Leishmaniasis is a common parasitic disease seen in many parts of the world, especially in areas where current U.S. and international forces are deployed. Approximately 350 million people are thought to be at risk of cutaneous leishmaniasis (CL) with an annual incidence of 1.5 million cases. Over 90% of cutaneous infections with Leishmania occur in the Middle East, Brazil, and Peru. Outbreaks of CL may occur in military personnel deployed to endemic areas. Since the incubation period for symptomatic CL ranges from weeks to months, symptoms may not appear until well after returning to the United States. As operations continue to expand globally, the exposure and concern for leishmaniasis persists for military physicians. We describe localized CL in a previously healthy male in an effort to help medical personnel identify leishmaniasis on the basis of cutaneous lesions alone, as well as increase diagnostic suspicion when treating patients in nonendemic areas. RESULTS: A previously healthy 30-year-old Saudi Arabian male presented to the emergency department with a 1-month history of four well-demarcated nonhealing, painless ulcers on his left ear, hand, and foot. Symptoms began shortly after arriving in the United States. He had been treated with trimethoprim/sulfamethoxazole, oral clindamycin, mupirocin ointment, and vancomycin for suspected infection without improvement of lesions. Upon presentation to dermatology, physical examination revealed a firm erythematous plaque with central ulceration on his left ear. Two shallow indurated ulcers were also found on his left fourth dorsal finger and left dorsal foot. Biopsy of the foot revealed granulomatous inflammation with predominantly lymphoplasmacytic infiltrate and multinucleated giant cells. Parasitized histiocytes were identified on hematoxylin and eosin stain and focally on Giemsa stain. Polymerase chain was consistent with a diagnosis of leishmaniasis and outpatient treatment was initiated with fluconazole 200 mg daily for 6 weeks. At 2-week follow-up, lesions were noted to be stabilized. DISCUSSION: CL has a wide variety of presentations. The classic lesion appears as a papule that will enlarge, often developing into a nodule or plaque-like lesion with central ulceration. The lesion may be covered with an eschar or by fibrinous material. This presentation can mimic many disease processes resulting in an extensive differential diagnosis that includes bacterial, fungal, and viral infections, cutaneous malignancy, and insect bites. The clinical course, treatment options, response to therapy, and prognosis are all highly variable and dependent on the causative species. Local therapy options, oral systemic agents, and parenteral agents have all shown varying results in the treatment of leishmaniasis. The difficulty with standardizing treatment options for CL stems from the lack of well-controlled studies and the lack of standardized outcome measures. This deficiency in comparative studies of treatment hinders consensual recommendations. However, the choice of the correct therapy often depends on the experience of the clinician, burden of disease, preferences of patients, and cost-effectiveness considerations for the patient and/or the health care system.
Assuntos
Leishmaniose Cutânea/complicações , Leishmaniose Cutânea/diagnóstico , Militares , Adulto , Serviço Hospitalar de Emergência/organização & administração , Fluconazol/farmacologia , Fluconazol/uso terapêutico , Humanos , Leishmaniose Cutânea/fisiopatologia , Masculino , Reação em Cadeia da Polimerase/métodos , Arábia Saudita/etnologia , Estados Unidos , CicatrizaçãoRESUMO
We evaluated the importance of neutrophils in the development of chronic lesions caused by L. Viannia spp. using the hamster as experimental model of American Cutaneous Leishmaniasis (ACL). Neutrophils infiltrated the lesion within the first six hours post-infection. Inhibition of this early infiltration using a polyclonal antibody or cyclophosphamide was associated with transient parasite control but the protective effect vanished when lesions became clinically apparent. At lesion onset (approximately 10 days p.i.), there was an increased proportion of both uninfected and infected macrophages, and subsequently a second wave of neutrophils infiltrated the lesion (after 19 days p.i.) This second neutrophil infiltration was associated with lesion necrosis and ulceration (R2 = 0.75) and maximum parasite burden. Intradermal delivery of N-formylmethionyl-leucyl-phenylalanine (fMLP), aimed to increase neutrophil infiltration, resulted in larger lesions with marked necrosis and higher parasite burden than in mock treated groups (p<0.001 each). In contrast, reduced neutrophil infiltration via cyclophosphamide-mediated depletion led to more benign lesions and lower parasite loads compared to controls (p<0.001 each). Neutrophils of the second wave expressed significantly lower GM-CSF, reactive oxygen species and nitric oxide than those of the first wave, suggesting that they had less efficient anti-leishmania activity. However, there was increased inflammatory cytokines and expression of neutrophil proteases (myeloperoxidase, cathepsin G and elastase) in lesions during the second wave of neutrophil infiltration compared with the levels reached during the first wave (6h p.i.). This suggests that augmented neutrophil proteases and inflammatory cytokines during the secondary wave of neutrophils could contribute to skin inflammation, ulceration and necrosis in ACL. The overall results indicate that neutrophils were unable to clear the infection in this model, and that the second wave of neutrophils played an important role in the severity of ACL.
Assuntos
Inflamação/sangue , Leishmaniose Cutânea/sangue , Necrose/sangue , Infiltração de Neutrófilos , Animais , Cricetinae , Modelos Animais de Doenças , Feminino , Humanos , Inflamação/parasitologia , Inflamação/fisiopatologia , Leishmania/patogenicidade , Leishmaniose Cutânea/parasitologia , Leishmaniose Cutânea/fisiopatologia , Macrófagos/patologia , Necrose/parasitologia , Necrose/fisiopatologia , Neutrófilos/patologia , Óxido Nítrico/metabolismo , Carga Parasitária , Espécies Reativas de Oxigênio/metabolismo , Estados UnidosRESUMO
Cutaneous leishmaniasis (CL) is the most common form of the leishmaniasis in humans. Ulcerative painless skin lesions are predominant clinical features of CL. Wider data indicate pain accompanies human leishmaniasis, out with areas of painless ulcerative lesions per se. In rodents, Leishmania (L.) major infection induces nociceptive behaviors that correlate with peripheral cytokine levels. However, the role of the spinal cord in pain processing after Leishmania infection has not been investigated. Balb/c mice received intraplantar (i.pl.) injection of Leishmania (L). amazonensis and hyperalgesia, edema, parasitism, and spinal cord TNFα, TNFR1 and TNFR2 mRNA expression, and NFκB activation were evaluated. The effects of intrathecal (i.t.) injection of morphine, TNFα, TNFα inhibitors (etanercept and adalimumab) and NFκB inhibitor (PDTC) were investigated. The present study demonstrates that Leishmania (L.) amazonensis infection in balb/c mice induces chronic mechanical and thermal hyperalgesia in an opioid-sensitive manner. Spinal cord TNFα mRNA expression increased in a time-dependent manner, peaking between 30 and 40 days after infection. At the peak of TNFα mRNA expression (day 30), there was a concomitant increase in TNFR1 and TNFR2 mRNA expression. TNFα i.t. injection enhanced L. (L.) amazonensis-induced hyperalgesia. Corroborating a role for TNFα in L. (L.) amazonensis-induced hyperalgesia, i.t. treatment with the TNFα inhibitors, etanercept and adalimumab inhibited the hyperalgesia. L. (L.) amazonensis also induced spinal cord activation of NFκB, and PDTC (given i.t.), also inhibited L. (L.) amazonensis-induced hyperalgesia, and spinal cord TNFα, TNFR1 and TNFR2 mRNA expression. Moreover, L. (L.) amazonensis-induced spinal cord activation of NFκB was also inhibited by etanercept and adalimumab as well as PDTC i.t. TREATMENT: These results demonstrate that endogenous spinal cord TNFα and NFκB activation contribute to L. (L.) amazonensis-induced hyperalgesia in mice. Thus, spinal cord TNFα and NFκB are potential therapeutic targets for Leishmania infection-induced pain.
Assuntos
Hiperalgesia/parasitologia , Leishmania mexicana/fisiologia , Leishmaniose Cutânea/parasitologia , NF-kappa B/metabolismo , Medula Espinal/fisiopatologia , Fator de Necrose Tumoral alfa/metabolismo , Adalimumab/administração & dosagem , Adalimumab/uso terapêutico , Animais , Etanercepte/administração & dosagem , Etanercepte/uso terapêutico , Hiperalgesia/tratamento farmacológico , Hiperalgesia/metabolismo , Hiperalgesia/fisiopatologia , Leishmaniose Cutânea/tratamento farmacológico , Leishmaniose Cutânea/fisiopatologia , Masculino , Camundongos Endogâmicos BALB C , Morfina/uso terapêutico , NF-kappa B/antagonistas & inibidores , NF-kappa B/genética , Carga Parasitária , Pirrolidinas/administração & dosagem , Pirrolidinas/uso terapêutico , RNA Mensageiro/metabolismo , Receptores Tipo I de Fatores de Necrose Tumoral/genética , Receptores Tipo I de Fatores de Necrose Tumoral/metabolismo , Receptores Tipo II do Fator de Necrose Tumoral/genética , Receptores Tipo II do Fator de Necrose Tumoral/metabolismo , Medula Espinal/metabolismo , Tiocarbamatos/administração & dosagem , Tiocarbamatos/uso terapêutico , Fator de Necrose Tumoral alfa/administração & dosagem , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/genéticaRESUMO
BACKGROUND: The treatment for leishmaniasis is currently based on pentavalent antimonials and amphotericin B; however, these drugs result in numerous adverse side effects. The lack of affordable therapy has necessitated the urgent development of new drugs that are efficacious, safe, and more accessible to patients. Natural products are a major source for the discovery of new and selective molecules for neglected diseases. In this paper, we evaluated the effect of apigenin on Leishmania amazonensis in vitro and in vivo and described the mechanism of action against intracellular amastigotes of L. amazonensis. METHODOLOGY/PRINCIPAL FINDING: Apigenin reduced the infection index in a dose-dependent manner, with IC50 values of 4.3 µM and a selectivity index of 18.2. Apigenin induced ROS production in the L. amazonensis-infected macrophage, and the effects were reversed by NAC and GSH. Additionally, apigenin induced an increase in the number of macrophages autophagosomes after the infection, surrounding the parasitophorous vacuole, suggestive of the involvement of host autophagy probably due to ROS generation induced by apigenin. Furthermore, apigenin treatment was also effective in vivo, demonstrating oral bioavailability and reduced parasitic loads without altering serological toxicity markers. CONCLUSIONS/SIGNIFICANCE: In conclusion, our study suggests that apigenin exhibits leishmanicidal effects against L. amazonensis-infected macrophages. ROS production, as part of the mechanism of action, could occur through the increase in host autophagy and thereby promoting parasite death. Furthermore, our data suggest that apigenin is effective in the treatment of L. amazonensis-infected BALB/c mice by oral administration, without altering serological toxicity markers. The selective in vitro activity of apigenin, together with excellent theoretical predictions of oral availability, clear decreases in parasite load and lesion size, and no observed compromises to the overall health of the infected mice encourage us to supports further studies of apigenin as a candidate for the chemotherapeutic treatment of leishmaniasis.
Assuntos
Antipruriginosos/administração & dosagem , Apigenina/administração & dosagem , Autofagia/efeitos dos fármacos , Leishmania/fisiologia , Leishmaniose Cutânea/tratamento farmacológico , Leishmaniose Cutânea/fisiopatologia , Espécies Reativas de Oxigênio/imunologia , Animais , Avaliação Pré-Clínica de Medicamentos , Feminino , Humanos , Leishmania/efeitos dos fármacos , Leishmaniose Cutânea/imunologia , Leishmaniose Cutânea/parasitologia , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB CRESUMO
Post-kala-azar dermal leishmaniasis (PKDL) is an important factor in kala-azar transmission; hence its early detection and assessment of effective treatment is very important for disease control. In present study on 60 PKDL cases presented with macular, mixed papulonodular, or erythematous lesions, Leishmania parasites were demonstrated microscopically in 91% of papulonodular and 40% of macular lesions. Cellular infiltrates in skin biopsy imprint smears from lesions were mononuclear cells, 25-300/OIF (oil immersion field), predominantly histiocytes with vacuolation, many lymphocytes, some plasma cells, and Leishmania amastigotes 0-20/OIF. Cases with no demonstrable parasites were diagnosed on the basis of past history of VL, lesion's distribution, cytopathological changes, and positive DAT (86.83%). Following antileishmanial treatment with SAG, papulonodular forms of PKDL lesions disappeared clinically but microscopically the mononuclear cells (20-200/OIF) persisted in the dermal lesions. Response observed in macular PKDL lesions was poor which persisted both clinically and cytopathologically. Follow-up of PKDL will assess the effectivity of treatment as either disappearance of lesions or any relapse. Studies on involvement of immunological factors, that is, certain cytokines (IL-10, TGF-ß, etc.) and chemokines (macrophage inflammatory protein, MIP 1-α, etc.) in PKDL, may provide insight for any role in the treatment response.
Assuntos
Leishmaniose Cutânea/fisiopatologia , Leishmaniose Visceral/imunologia , Leishmaniose Visceral/fisiopatologia , Adolescente , Adulto , Criança , Feminino , Humanos , Interleucina-10/imunologia , Leishmania donovani/imunologia , Leishmania donovani/patogenicidade , Leishmaniose Cutânea/imunologia , Leishmaniose Cutânea/transmissão , Leishmaniose Visceral/transmissão , Masculino , Pessoa de Meia-Idade , Fator de Crescimento Transformador beta/imunologia , Resultado do TratamentoAssuntos
Anfotericina B , Leishmania , Leishmaniose Cutânea , Úlcera/diagnóstico , Adulto , Anfotericina B/administração & dosagem , Anfotericina B/efeitos adversos , Antiprotozoários/administração & dosagem , Antiprotozoários/efeitos adversos , Braço/patologia , Biópsia , Diagnóstico Diferencial , Antebraço/patologia , Humanos , Leishmania/efeitos dos fármacos , Leishmania/isolamento & purificação , Leishmaniose Cutânea/diagnóstico , Leishmaniose Cutânea/tratamento farmacológico , Leishmaniose Cutânea/etiologia , Leishmaniose Cutânea/fisiopatologia , Masculino , Parede Torácica/patologia , Resultado do TratamentoRESUMO
Leishmaniasis is an infectious disease caused by protozoa of the genus Leishmania transmitted by insects of the genus Lutzomyia sp. or Phlebotomus sp. The main syndromes are cutaneous leishmaniasis, mucocutaneous leishmaniasis, visceral leishmaniasis (kala-azar) and post-kala-azar dermal leishmaniasis. This article reviews kidney involvement in cutaneous and visceral leishmaniasis, highlighting the aspects of their pathophysiology, clinical manifestations, histopathological findings, outcome and treatment.
Assuntos
Humanos , Nefropatias/parasitologia , Leishmaniose Cutânea/complicações , Leishmaniose Visceral/complicações , Nefropatias/patologia , Nefropatias/fisiopatologia , Leishmaniose Cutânea/patologia , Leishmaniose Cutânea/fisiopatologia , Leishmaniose Visceral/patologia , Leishmaniose Visceral/fisiopatologiaAssuntos
Antiprotozoários/uso terapêutico , Úlcera da Perna/diagnóstico , Úlcera da Perna/tratamento farmacológico , Leishmaniose Cutânea/diagnóstico , Leishmaniose Cutânea/tratamento farmacológico , Meglumina/uso terapêutico , Compostos Organometálicos/uso terapêutico , Cicatrização/fisiologia , Biópsia , Diagnóstico Diferencial , Feminino , Humanos , Irã (Geográfico) , Úlcera da Perna/etiologia , Úlcera da Perna/fisiopatologia , Leishmaniose Cutânea/complicações , Leishmaniose Cutânea/fisiopatologia , Antimoniato de Meglumina , Pessoa de Meia-IdadeRESUMO
Disease progression in response to infection can be strongly influenced by both pathogen burden and infection-induced immunopathology. While current therapeutics focus on augmenting protective immune responses, identifying therapeutics that reduce infection-induced immunopathology are clearly warranted. Despite the apparent protective role for murine CD8⺠T cells following infection with the intracellular parasite Leishmania, CD8⺠T cells have been paradoxically linked to immunopathological responses in human cutaneous leishmaniasis. Transcriptome analysis of lesions from Leishmania braziliensis patients revealed that genes associated with the cytolytic pathway are highly expressed and CD8⺠T cells from lesions exhibited a cytolytic phenotype. To determine if CD8⺠T cells play a causal role in disease, we turned to a murine model. These studies revealed that disease progression and metastasis in L. braziliensis infected mice was independent of parasite burden and was instead directly associated with the presence of CD8⺠T cells. In mice with severe pathology, we visualized CD8⺠T cell degranulation and lysis of L. braziliensis infected cells. Finally, in contrast to wild-type CD8⺠T cells, perforin-deficient cells failed to induce disease. Thus, we show for the first time that cytolytic CD8⺠T cells mediate immunopathology and drive the development of metastatic lesions in cutaneous leishmaniasis.
Assuntos
Citotoxicidade Imunológica , Modelos Animais de Doenças , Leishmania braziliensis/imunologia , Leishmaniose Cutânea/imunologia , Pele/imunologia , Linfócitos T Citotóxicos/imunologia , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Brasil , Progressão da Doença , Perfilação da Expressão Gênica , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Humanos , Leishmania braziliensis/isolamento & purificação , Leishmaniose Cutânea/parasitologia , Leishmaniose Cutânea/patologia , Leishmaniose Cutânea/fisiopatologia , Leishmaniose Tegumentar Difusa/etiologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Pele/parasitologia , Pele/patologia , Organismos Livres de Patógenos Específicos , Linfócitos T Citotóxicos/parasitologia , Linfócitos T Citotóxicos/patologiaRESUMO
Obligate intracellular pathogens such as Leishmania specifically target host phagocytes for survival and replication. Phosphoinositide 3-kinase γ (PI3Kγ), a member of the class I PI3Ks that is highly expressed by leukocytes, controls cell migration by initiating actin polymerization and cytoskeletal reorganization, which are processes also critical for phagocytosis. In this study, we demonstrate that class IB PI3K, PI3Kγ, plays a critical role in pathogenesis of chronic cutaneous leishmaniasis caused by L. mexicana. Using the isoform-selective PI3Kγ inhibitor, AS-605240 and PI3Kγ gene-deficient mice, we show that selective blockade or deficiency of PI3Kγ significantly enhances resistance against L. mexicana that is associated with a significant suppression of parasite entry into phagocytes and reduction in recruitment of host phagocytes as well as regulatory T cells to the site of infection. Furthermore, we demonstrate that AS-605240 is as effective as the standard antileishmanial drug sodium stibogluconate in treatment of cutaneous leishmaniasis caused by L. mexicana. These findings reveal a unique role for PI3Kγ in Leishmania invasion and establishment of chronic infection, and demonstrate that therapeutic targeting of host pathways involved in establishment of infection may be a viable strategy for treating infections caused by obligate intracellular pathogens such as Leishmania.
Assuntos
Resistência à Doença/efeitos dos fármacos , Leishmania mexicana , Leishmaniose Cutânea/parasitologia , Fosfatidilinositol 3-Quinases/metabolismo , Quinoxalinas/farmacologia , Tiazolidinedionas/farmacologia , Animais , Gluconato de Antimônio e Sódio/uso terapêutico , Citometria de Fluxo , Interações Hospedeiro-Parasita/efeitos dos fármacos , Humanos , Leishmaniose Cutânea/fisiopatologia , Macrófagos , Camundongos , Camundongos Endogâmicos C57BL , Microscopia de Fluorescência , Neutrófilos , Fagócitos/efeitos dos fármacos , Inibidores de Fosfoinositídeo-3 Quinase , Quinoxalinas/uso terapêutico , Tiazolidinedionas/uso terapêuticoRESUMO
Leishmania (Viannia) shawi was recently characterized and few studies concerning modifications in cellular and humoral immune responses in experimental leishmaniasis have been conducted. In this work, immunopathological changes induced by L. shawi in chronically infected BALB/c mice were investigated. Infected BALB/c mice developed increased lesion size associated with strong inflammatory infiltrate diffusely distributed in the dermis, with highly infected macrophages. The humoral immune response was predominantly directed toward the IgG1 isotype. The functional activity of CD4(+) and CD8(+) T cells showed significantly increased TNF-alpha mRNA levels associated with reduced IFN-gamma expression by CD4(+) T cells and the double negative (dn) CD4CD8 cell subset. High IL-4 levels expressed by CD8(+) T cells and dnCD4CD8 and TGF-beta by CD4(+) and CD8(+) T cells were detected, while IL-10 was highly expressed by all three cell subpopulations. Taken together, these results show an evident imbalance between TNF-alpha and IFN-gamma that is unfavorable to amastigote replication control. Furthermore, L. shawi seems to regulate different cell populations to express deactivating cytokines to avoid its own destruction. This study indicates BALB/c mice as a potentially good experimental model for further studies on American cutaneous leishmaniosis caused by L. shawi.
Assuntos
Anticorpos Antiprotozoários/sangue , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Modelos Animais de Doenças , Leishmania , Leishmaniose Cutânea , Animais , Citocinas/metabolismo , Derme/imunologia , Derme/parasitologia , Derme/patologia , Humanos , Imunoglobulina G/sangue , Leishmania/classificação , Leishmania/crescimento & desenvolvimento , Leishmania/imunologia , Leishmania/patogenicidade , Leishmaniose Cutânea/imunologia , Leishmaniose Cutânea/parasitologia , Leishmaniose Cutânea/patologia , Leishmaniose Cutânea/fisiopatologia , Macrófagos/parasitologia , Camundongos , Camundongos Endogâmicos BALB CRESUMO
Indian post-kala-azar dermal leishmaniasis (PKDL) is a low-frequency (5-10%) dermal sequela of visceral leishmaniasis (VL) caused by Leishmania donovani; importantly, affected individuals are speculated to be parasite reservoirs. Insight into its immunopathogenesis could translate into rational immunomodulatory therapeutic approaches against leishmaniases. In patients with PKDL (n=21), peripheral lymphocytes were analyzed for surface markers, intracellular cytokines, and lymphoproliferative responses using flow cytometry. In lesional tissue biopsies (n=12), expression of counter-regulatory cytokines (IFN-gamma and IL-10) and the T-regulatory transcription factor forkhead box protein 3 (Foxp3) was analyzed using reverse transcriptase-PCR, along with immunohistochemical detection (n=8) of CD3 and Foxp3 positivity. In patients with PKDL, circulating CD8(+)CD28(-) and antigen-induced IL-10(+)CD3(+) lymphocytes were increased and receded with treatment. CD8(+) lymphocytes showed impaired proliferative responses to L. donovani antigen (LDA) and phytohemagglutinin, which were reinstated after treatment. At presentation, the upregulated lesional IFN-gamma and IL-10 messenger RNA (mRNA), Foxp3 mRNA, and protein were curtailed after treatment. In Indian patients with PKDL, increased frequency of the CD8(+)CD28(-) phenotype, enhanced antigen-specific IL-10 production, and accompanying anergy of circulating lymphocytes suggest their regulatory nature. Furthermore, the concomitantly elevated lesional expression of Foxp3 suggests their possible recruitment into the lesional site, which would sustain disease pathology.
Assuntos
Fatores de Transcrição Forkhead , Leishmania donovani/imunologia , Leishmaniose Cutânea/imunologia , Leishmaniose Visceral/imunologia , Linfócitos T Reguladores/fisiologia , Linfócitos T Reguladores/parasitologia , Adulto , Antígenos de Protozoários/imunologia , Antígenos CD28/metabolismo , Complexo CD3/metabolismo , Antígenos CD8/metabolismo , Linfócitos T CD8-Positivos/parasitologia , Linfócitos T CD8-Positivos/fisiologia , Derme/imunologia , Derme/parasitologia , Derme/patologia , Feminino , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/imunologia , Fatores de Transcrição Forkhead/metabolismo , Humanos , Índia , Interferon gama/metabolismo , Interleucina-10/metabolismo , Leishmaniose Cutânea/patologia , Leishmaniose Cutânea/fisiopatologia , Leishmaniose Visceral/patologia , Leishmaniose Visceral/fisiopatologia , Masculino , Proteínas de Membrana/metabolismo , Pessoa de Meia-Idade , RNA Mensageiro/metabolismoRESUMO
Intracellular parasites of the genus Leishmania generate severe diseases in humans, which are associated with a failure of the infected host to induce a protective interferon gamma (IFNgamma)-mediated immune response. We tested the role of the JAK/STAT1 signaling pathway in Leishmania pathogenesis by utilizing knockout mice lacking the signal transducer and activator of transcription 1 (Stat1) and derived macrophages. Unexpectedly, infection of Stat1-deficient macrophages in vitro with promastigotes from Leishmania major and attenuated LPG1 knockout mutants (lpg(-)) specifically lacking lipophosphoglycan (LPG) resulted in a twofold increased intracellular growth, which was independent of IFNgamma and associated with a substantial increase in phagosomal pH. Phagosomes in Stat1-/- macrophages showed normal maturation as judged by the accumulation of the lysosomal marker protein rab7, and provided normal vATPase activity, but were defective in the anion conductive pathway required for full vesicular acidification. Our results suggest a role of acidic pH in the control of intracellular Leishmania growth early during infection and identify for the first time an unexpected role of Stat1 in natural anti-microbial resistance independent from its function as IFNgamma-induced signal transducer. This novel Stat1 function may have important implications to studies of other pathogens, as the acidic phagolysosomal pH plays an important role in antigen processing and the uncoating process of many viruses.
Assuntos
Leishmania major/patogenicidade , Fagossomos/fisiologia , Fator de Transcrição STAT1/fisiologia , Animais , Fusão Celular , Feminino , Glicoesfingolipídeos/deficiência , Concentração de Íons de Hidrogênio , Interferon gama/fisiologia , Leishmaniose Cutânea/imunologia , Leishmaniose Cutânea/fisiopatologia , Lisossomos/fisiologia , Macrófagos/microbiologia , Camundongos , Fator de Transcrição STAT1/deficiênciaRESUMO
Local inflammation during cutaneous leishmaniasis is accompanied by accumulation of CD11b(+) cells at the site of the infection. A functional role for these monocytic cells in local angiogenesis in leishmaniasis has not been described so far. Here, we show that CD11b(+) cells express high levels of the myeloid differentiation antigen carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1). In experimental cutaneous leishmaniasis in C57BL/6 wild-type (B6.WT) and B6.Ceacam1(-/-) mice, we found that only B6.Ceacam1(-/-) mice develop edemas and exhibit impairment of both hemangiogenesis and lymphangiogenesis. Because CEACAM1 expression correlates with functional angiogenesis, we further analyzed the role of the CD11b(+) population. In B6.Ceacam1(-/-) mice, we found systemic reduction of Ly-6C(high)/CD11b(high) monocyte precursors. To investigate whether CEACAM1(+) myeloid cells are causally related to efficient angiogenesis, we used reverse bone marrow transplants (BMTs) to restore CEACAM1(+) or CEACAM1(-) bone marrow in B6.Ceacam1(-/-) or B6.WT recipients, respectively. We found that angiogenesis was restored by CEACAM1(+) BMT only. In addition, we observed reduced morphogenic potential of inflammatory cells in Matrigel implants in CEACAM1(-) backgrounds or after systemic depletion of CD11b(high) macrophages. Taken together, we show for the first time that CEACAM1(+) myeloid cells are crucial for angiogenesis in inflammation.
Assuntos
Antígeno Carcinoembrionário/análise , Inflamação/fisiopatologia , Leishmaniose Cutânea/fisiopatologia , Células Mieloides/fisiologia , Neovascularização Patológica/fisiopatologia , Animais , Anticorpos Antiprotozoários/biossíntese , Transplante de Medula Óssea , Antígeno CD11b/análise , Antígeno Carcinoembrionário/biossíntese , Antígeno Carcinoembrionário/genética , Colágeno , Combinação de Medicamentos , Edema/etiologia , Edema/patologia , Glicoproteínas/biossíntese , Imunidade Celular , Implantes Experimentais , Inflamação/etiologia , Inflamação/imunologia , Interferon gama/biossíntese , Laminina , Leishmania major/imunologia , Leishmaniose Cutânea/complicações , Leishmaniose Cutânea/imunologia , Leishmaniose Cutânea/patologia , Vasos Linfáticos/metabolismo , Macrófagos/parasitologia , Macrófagos/fisiologia , Proteínas de Membrana Transportadoras , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Células Mieloides/química , Células Mieloides/classificação , Neovascularização Patológica/patologia , Proteoglicanas , Quimera por Radiação , Células Th1/imunologiaRESUMO
Leishmaniasis occurs as a spectrum of clinical syndromes divided into cutaneous, mucocutaneous and visceral forms. The epidemiology and clinical features are highly variable owing to the interplay of many factors ranging from parasite species and strains, vectors, host genetics and environment. Currently, there is no effective licensed vaccine for use in humans against leishmaniasis. Most traditional and low-cost treatment options, particularly in poor and endemic areas, are toxic with many adverse reactions and they require a long course of administration. The use of more effective, less toxic drugs is limited because total treatment cost is very high (expensive) and there are fears of development of drug resistance. Recent studies indicate that certain strategies aimed at modulating the host immune response (collectively called immunotherapy) could result in prophylactic and/or therapeutic cure of leishmaniasis under both laboratory and field conditions. In this review, we focus on treatment of leishmaniasis with a particular emphasis on immunotherapy/immunochemotherapy as an alternative to conventional drug treatment.
Assuntos
Antígenos de Protozoários/metabolismo , Citocinas/biossíntese , Imunoterapia , Leishmania/imunologia , Leishmaniose Cutânea/terapia , Animais , Anticorpos Monoclonais/uso terapêutico , Antígenos de Protozoários/genética , Antígenos de Protozoários/imunologia , Antígenos de Protozoários/uso terapêutico , Citocinas/genética , Citocinas/imunologia , Citocinas/metabolismo , Custos de Medicamentos , Humanos , Leishmania/crescimento & desenvolvimento , Leishmania/patogenicidade , Leishmaniose Cutânea/economia , Leishmaniose Cutânea/imunologia , Leishmaniose Cutânea/fisiopatologia , Ativação Linfocitária/efeitos dos fármacos , Ativação Linfocitária/imunologia , Equilíbrio Th1-Th2/efeitos dos fármacos , Vacinas de DNARESUMO
Neutrophils are involved in the initial steps of most responses to pathogens. In the present study, we evaluated the effects of the interaction of apoptotic vs. necrotic human neutrophils on macrophage infection by Leishmania amazonensis. Phagocytosis of apoptotic, but not viable, neutrophils by Leishmania-infected macrophages led to an increase in parasite burden via a mechanism dependent on TGF-beta1 and PGE2. Conversely, infected macrophages' uptake of necrotic neutrophils induced killing of L. amazonensis. Leishmanicidal activity was dependent on TNF-alpha and neutrophilic elastase. Nitric oxide was not involved in the killing of parasites, but the interaction of necrotic neutrophils with infected macrophages resulted in high superoxide production, a process reversed by catalase, an inhibitor of reactive oxygen intermediate production. Initial events after Leishmania infection involve interactions with neutrophils; we demonstrate that phagocytosis of these cells in an apoptotic or necrotic stage can influence the outcome of infection, driving either parasite survival or destruction.
Assuntos
Leishmaniose Cutânea/fisiopatologia , Macrófagos/parasitologia , Neutrófilos/imunologia , Neutrófilos/fisiologia , Animais , Apoptose , Catalase/farmacologia , Efeitos Psicossociais da Doença , Dinoprostona/fisiologia , Humanos , Leishmania mexicana/patogenicidade , Leishmania mexicana/fisiologia , Leishmaniose Cutânea/patologia , Necrose , Neutrófilos/patologia , Fagocitose , Superóxidos/metabolismo , Fator de Crescimento Transformador beta1/fisiologiaAssuntos
Pé/patologia , Leishmaniose Cutânea/patologia , Leishmaniose Visceral/complicações , Pele/patologia , Animais , Gluconato de Antimônio e Sódio/uso terapêutico , Antiprotozoários/uso terapêutico , Biópsia por Agulha Fina , Diagnóstico Diferencial , Pé/parasitologia , Pé/fisiopatologia , Humanos , Leishmania/citologia , Leishmaniose Cutânea/parasitologia , Leishmaniose Cutânea/fisiopatologia , Leishmaniose Visceral/parasitologia , Leishmaniose Visceral/fisiopatologia , Macrófagos/parasitologia , Macrófagos/patologia , Masculino , Pessoa de Meia-Idade , Nepal , Valor Preditivo dos Testes , Pele/parasitologia , Pele/fisiopatologia , Resultado do TratamentoRESUMO
OBJECTIVE: Leishmania tropica is the causative agent of anthroponotic cutaneous leishmaniasis (CL) in Iran. The disease often heals within a year; however, the non-healing forms of disease are also known. The aim of the present study was the determination of the levels of soluble (s) CD26 and CD30 co-stimulatory molecules in sera of L. tropica-infected individuals. The correlations of sCD26 and sCD30 levels with clinical presentation of the disease were assessed. METHODS: The levels of sCD26 and sCD30 were determined by a sandwich enzyme-linked immunosorbent assay in sera from patients with acute and non-healing presentation of disease. RESULTS: The serum level of sCD26 was significantly higher in non-healing patients than in cases with acute CL (P<0.001). There was no significant difference in sCD26 level between patients with acute CL and healthy controls. However, the levels of sCD30 in sera from all L. tropica-infected individuals were higher than controls (P<0.001). A significant difference was also found in sCD30 level between non-healing cases and patients with acute CL (P<0.001). CONCLUSION: These findings suggest sCD30 is more relevant to clinical manifestation of cutaneous leishmaniasis than sCD26. The high sCD26 and sCD30 levels in non-healing patients reflect the presence of mixed Th1- and Th2-type responses in these patients.