Carregaro Vanessa, Valenzuela Jesus G, Cunha Thiago M, Verri Waldiceu A, Grespan Renata, Matsumura Graziela, Ribeiro José M C, Elnaiem Dia-Eldin, Silva João S, Cunha Fernando Q
Department of Biochemistry and Immunology, School of Medicine of Ribeirão Preto, University of Sao Paulo, Av Bandeirantes, 3900, Ribeirão Preto, SP, Brazil, 14049-900.
J Leukoc Biol. 2008 Jul;84(1):104-14. doi: 10.1189/jlb.1107797. Epub 2008 Apr 7.
In the present study, we investigated whether saliva from Phlebotomus papatasi and Phlebotomus duboscqi inhibited antigen-induced neutrophil migration and the mechanisms involved in these effects. The pretreatment of immunized mice with salivary gland extracts (SGE) of both phlebotomines inhibited OVA challenge-induced neutrophil migration and release of the neutrophil chemotactic mediators, MIP-1alpha, TNF-alpha, and leukotriene B4 (LTB4). Furthermore, SGE treatment enhanced the production of anti-inflammatory mediators, IL-10 and PGE2. SGE treatments failed to inhibit neutrophil migration and MIP-1alpha and LTB4 production in IL-10-/- mice, also failing in mice treated with nonselective (indomethacin) or selective (rofecoxibe) cyclooxygenase (COX) inhibitors. COX inhibition resulted in diminished SGE-induced IL-10 production, and PGE2 release triggered by SGE remained increased in IL-10-/- mice, suggesting that prostanoids are acting through an IL-10-dependent mechanism. SGE treatments in vivo reduced the OVA-induced lymphoproliferation of spleen-derived cells. Further, the in vitro incubation of bone marrow-derived dendritic cells (DC) with SGE inhibited the proliferation of CD4+T cells from OVA-immunized mice, which was reversed by indomethacin and anti-IL-10 antibody treatments. Supporting these results, SGE induced the production of PGE2 and IL-10 by DC, which were blocked by COX inhibition. These effects were associated with the reduction of DC-membrane expression of MHC-II and CD86 by SGE treatment. Altogether, the results showed that Phlebotomine saliva inhibits immune inflammation-induced neutrophil migration by an autocrine DC sequential production of PGE2/IL-10, suggesting that the saliva constituents might be promising therapeutic molecules to target immune inflammatory diseases.
在本研究中，我们调查了巴氏白蛉和杜氏白蛉的唾液是否抑制抗原诱导的中性粒细胞迁移以及这些作用所涉及的机制。用这两种白蛉的唾液腺提取物（SGE）预处理免疫小鼠，可抑制卵清蛋白攻击诱导的中性粒细胞迁移以及中性粒细胞趋化介质、巨噬细胞炎性蛋白-1α（MIP-1α）、肿瘤坏死因子-α（TNF-α）和白三烯B4（LTB4）的释放。此外，SGE处理增强了抗炎介质白细胞介素-10（IL-10）和前列腺素E2（PGE2）的产生。SGE处理未能抑制IL-10基因敲除小鼠的中性粒细胞迁移以及MIP-1α和LTB4的产生，在用非选择性（吲哚美辛）或选择性（罗非昔布）环氧化酶（COX）抑制剂处理的小鼠中也失败了。COX抑制导致SGE诱导的IL-10产生减少，而在IL-10基因敲除小鼠中，SGE触发的PGE2释放仍然增加，这表明前列腺素是通过一种依赖IL-10的机制发挥作用。SGE在体内处理降低了卵清蛋白诱导的脾源性细胞的淋巴细胞增殖。此外，用SGE体外培养骨髓来源的树突状细胞（DC）抑制了来自卵清蛋白免疫小鼠的CD4⁺T细胞的增殖，吲哚美辛和抗IL-10抗体处理可逆转这种抑制作用。支持这些结果的是，SGE诱导DC产生PGE2和IL-10，而COX抑制可阻断这些产生。这些作用与SGE处理降低DC膜上MHC-II和CD86的表达有关。总之，结果表明白蛉唾液通过DC自分泌顺序产生PGE2/IL-10来抑制免疫炎症诱导的中性粒细胞迁移，这表明唾液成分可能是治疗免疫炎症性疾病的有前景的治疗分子。
