Department of Pharmacology, Faculty of Medicine of Ribeirão Preto, University of Sao Paulo, Ribeirão Preto, SP, Brazil.
Int Immunopharmacol. 2012 Apr;12(4):603-10. doi: 10.1016/j.intimp.2012.02.004. Epub 2012 Feb 21.
Several studies have pointed out the immunomodulatory properties of the Salivary Gland Extract (SGE) from Lutzomyia longipalpis. We aimed to identify the SGE component (s) responsible for its effect on ovalbumin (OVA)-induced neutrophil migration (NM) and to evaluate the effect of SGE and components in the antigen-induced arthritis (AIA) model. We tested the anti-arthritic activities of SGE and the recombinant LJM111 salivary protein (rLJM111) by measuring the mechanical hypernociception and the NM into synovial cavity. Furthermore, we measured IL-17, TNF-α and IFN-γ released by lymph nodes cells stimulated with mBSA or anti-CD3 using enzyme-linked immunosorbent assay (ELISA). Additionally, we tested the effect of SGE and rLJM111 on co-stimulatory molecules expression (MHC-II and CD-86) by flow cytometry, TNF-α and IL-10 production (ELISA) of bone marrow-derived dendritic cells (BMDCs) stimulated with LPS, chemotaxis and actin polymerization from neutrophils. Besides, the effect of SGE on CXCR2 and GRK-2 expression on neutrophils was investigated. We identified one plasmid expressing the protein LJM111 that prevented NM in OVA-challenged immunized mice. Furthermore, both SGE and rLJM111 inhibited NM and pain sensitivity in AIA and reduced IL-17, TNF-α and IFN-γ. SGE and rLJM111 also reduced MHC-II and CD-86 expression and TNF-α whereas increased IL-10 release by LPS-stimulated BMDCs. SGE, but not LJM 111, inhibited neutrophils chemotaxis and actin polymerization. Additionally, SGE reduced neutrophil CXCR2 expression and increased GRK-2. Thus, rLJM111 is partially responsible for SGE mechanisms by diminishing DC function and maturation but not chemoattraction of neutrophils.
几项研究指出了唾液腺提取物（SGE）对长角血蜱唾液腺提取物（Lutzomyia longipalpis）的免疫调节特性。我们旨在确定负责其对卵清蛋白（OVA）诱导的中性粒细胞迁移（NM）作用的 SGE 成分（s），并评估 SGE 和成分在抗原诱导的关节炎（AIA）模型中的作用。我们通过测量机械性痛觉过敏和 NM 进入滑膜腔来测试 SGE 和重组 LJM111 唾液蛋白（rLJM111）的抗关节炎活性。此外，我们通过酶联免疫吸附试验（ELISA）测量刺激 mBSA 或抗 CD3 后淋巴结细胞释放的 IL-17、TNF-α 和 IFN-γ，来测试 SGE 和 rLJM111 对共刺激分子表达（MHC-II 和 CD-86）的影响，并用 LPS 刺激骨髓来源的树突状细胞（BMDCs），用流式细胞术测量 TNF-α 和 IL-10 的产生（ELISA），并测量中性粒细胞的趋化作用和肌动蛋白聚合作用。此外，还研究了 SGE 对中性粒细胞中 CXCR2 和 GRK-2 表达的影响。我们确定了一个表达蛋白 LJM111 的质粒，该质粒可防止 OVA 挑战免疫小鼠的 NM。此外，SGE 和 rLJM111 均可抑制 AIA 中的 NM 和疼痛敏感性，并降低 IL-17、TNF-α 和 IFN-γ。SGE 和 rLJM111 还降低了 LPS 刺激的 BMDCs 中 MHC-II 和 CD-86 的表达和 TNF-α，而增加了 IL-10 的释放。SGE 可抑制中性粒细胞趋化作用和肌动蛋白聚合作用，但 LJM111 则不然。此外，SGE 降低了中性粒细胞 CXCR2 的表达，增加了 GRK-2 的表达。因此，rLJM111 部分通过降低 DC 功能和成熟度而不是中性粒细胞的趋化作用来负责 SGE 机制，但不负责中性粒细胞的趋化作用。
