Ahmed A A, Wahbi A, Nordlind K, Kharazmi A, Sundqvist K G, Mutt V, Lidén S
Department of Dermatology, Karolinska Hospital, Stockholm, Sweden.
Scand J Immunol. 1998 Jul;48(1):79-85. doi: 10.1046/j.1365-3083.1998.00380.x.
Recruitment, migration and adherence of macrophages and their interaction with inoculated promastigotes are key steps in the initiation of the inflammatory process in cutaneous leishmaniasis. Parasite- and nervous system-derived factors might be involved in this process. In the present study the chemotactic activities of live, killed and sonicated Leishmania major promastigotes and of the promastigote culture supernatant as well as the L. major surface protease gp63 towards a murine macrophage cell line, Raw 264.7, were investigated, using the Boyden technique. The sensory neuropeptides SOM, CGRP and SP, and the autonomic neuropeptides VIP and NPY, were also investigated for possible modulatory effects on this chemotaxis, using the living promastigotes. Living promastigotes were the most efficient attractants for macrophages compared with other forms of the parasites. Prior incubation of the macrophages with the parasites completely abolished the chemotactic activity. This might indicate that the living promastigote chemotaxis is a receptor-mediated process. On the other hand, paraformaldehyde-killed promastigotes not only failed to induce macrophage chemotaxis but also inhibited it in comparison with the control. The surface protease gp63 tended to inhibit the macrophage chemotactic activity and the sonicate tended to stimulate it compared with controls. The culture supernatant had no effect, indicating that the chemoattractive factors putatively synthesized by the living promastigotes are not released to the surrounding medium. Somatostatin inhibited L. major promastigote-induced macrophage migration at a high concentration, 10(-6) M, while substance P inhibited it at both low concentrations, 10(-10) and 10(-9) M, and a high one, 10(-6) M, the last-mentioned having the greatest inhibitory effect. A stimulatory effect of calcitonin gene-related peptide was found at high concentrations, 10(-5) and 10(-6) M. Vasoactive intestinal peptide stimulated macrophage chemotactic activity at both a high, 10(-5) M, and at a low, 10(-9) M, concentration, the same concentration at which neuropeptide Y exerted its maximum inhibitory effect.
巨噬细胞的募集、迁移和黏附以及它们与接种的前鞭毛体的相互作用是皮肤利什曼病炎症过程起始的关键步骤。寄生虫和神经系统衍生的因素可能参与了这一过程。在本研究中，使用博伊登技术研究了活的、杀死的和超声破碎的硕大利什曼原虫前鞭毛体、前鞭毛体培养上清液以及硕大利什曼原虫表面蛋白酶gp63对小鼠巨噬细胞系Raw 264.7的趋化活性。还使用活的前鞭毛体研究了感觉神经肽SOM、CGRP和SP以及自主神经肽VIP和NPY对这种趋化作用可能的调节作用。与寄生虫的其他形式相比，活的前鞭毛体是巨噬细胞最有效的趋化因子。巨噬细胞与寄生虫预先孵育完全消除了趋化活性。这可能表明活的前鞭毛体趋化是一个受体介导的过程。另一方面，与对照相比，多聚甲醛杀死的前鞭毛体不仅未能诱导巨噬细胞趋化，反而抑制了趋化。表面蛋白酶gp63倾向于抑制巨噬细胞趋化活性，与对照相比，超声破碎物倾向于刺激趋化活性。培养上清液没有作用，表明活的前鞭毛体可能合成的趋化因子没有释放到周围培养基中。生长抑素在高浓度10(-6)M时抑制硕大利什曼原虫前鞭毛体诱导的巨噬细胞迁移，而P物质在低浓度10(-10)和10(-9)M以及高浓度10(-6)M时均抑制巨噬细胞迁移，最后提到的浓度具有最大的抑制作用。在高浓度10(-5)和10(-6)M时发现降钙素基因相关肽有刺激作用。血管活性肠肽在高浓度10(-5)M和低浓度10(-9)M时均刺激巨噬细胞趋化活性，神经肽Y在相同浓度时发挥最大抑制作用。
