RESUMO
rIL-10 plays a major role in restricting exaggerated inflammatory and immune responses, thus preventing tissue damage. However, the restriction of inflammatory and immune responses by IL-10 can also favor the development and/or persistence of chronic infections or neoplasms. Dogs that succumb to canine leishmaniasis (CanL) caused by L. infantum develop exhaustion of T lymphocytes and are unable to mount appropriate cellular immune responses to control the infection. These animals fail to mount specific lymphoproliferative responses and produce interferon gamma and TNF-alpha that would activate macrophages and promote destruction of intracellular parasites. Blocking IL-10 signaling may contribute to the treatment of CanL. In order to obtain a tool for this blockage, the present work endeavored to identify the canine casIL-10R1 amino acid sequence, generate a recombinant baculovirus chromosome encoding this molecule, which was expressed in insect cells and subsequently purified to obtain rcasIL-10R1. In addition, rcasIL-10R1 was able to bind to homologous IL-10 and block IL-10 signaling pathway, as well as to promote lymphoproliferation in dogs with leishmaniasis caused by L. infantum.
Assuntos
Interleucina-10/metabolismo , Leishmaniose/tratamento farmacológico , Receptores de Interleucina-10/metabolismo , Animais , Linhagem Celular , Citocinas/metabolismo , Doenças do Cão/genética , Cães , Feminino , Imunidade Celular/imunologia , Imunidade Celular/fisiologia , Interferon gama/genética , Interleucina-10/agonistas , Interleucina-12/genética , Leishmania infantum/imunologia , Leishmania infantum/patogenicidade , Leishmaniose/imunologia , Macrófagos/metabolismo , Masculino , Camundongos , Receptores de Interleucina-10/efeitos dos fármacos , Transdução de Sinais , Linfócitos T/imunologia , Fator de Necrose Tumoral alfaRESUMO
OBJECTIVE: To determine whether high levels of interleukin (IL)-10 can attenuate the production of tumor necrosis factor (TNF)-alpha-induced proinflammatory cytokines in endometriotic stromal cells. DESIGN: Prospective study. SETTING: Department of Ob/Gyn, Tottori University, Japan. PATIENT(S): Thirty-five patients with ovarian endometrioma and ten patients with uterine myoma. INTERVENTION(S): Endometriotic stromal cells were obtained from chocolate cyst linings of ovaries. Endometrial stromal cells obtained from patient with uterine myoma. MAIN OUTCOME MEASURE(S): Expression of IL-10 gene in endometriotic or endometrial stromal cells was determined by real-time reverse-transcriptase polymerase chain reaction (RT-PCR). We performed immunohistochemical staining to find the presence of IL-10 and IL-10 receptors 1 and 2. We examined the effects of TNF-alpha and IL-10 on the expression of IL-6 or IL-8 by real-time RT-PCR and ELISA. We examined the activation of intracellular signal transduction molecules in endometriotic stromal cells by Western blotting. RESULT(S): Addition of IL-10 suppressed the expressions of IL-6 induced by TNF-alpha and IL-10 induced the phosphorylation of STAT3 in endometriotic stromal cells. TNF-alpha induced the expression of phosphorylated ERK1/2, JNK1/2, and I kappaB. Adding IL-10 suppressed the phosphorylation of these signal molecules. CONCLUSION(S): Interleukin-10 attenuates TNF-alpha-induced IL-6 synthesis via NF-kappaB and MAPK pathways in endometriotic cells.. Interleukin-10 may play a significant role in the pathogenesis of endometriosis.
