Molecular Systematics Laboratory, Parasitology Department, Pasteur Institute of Iran, Tehran, Iran.
Parasitology Department, Pasteur Institute of Iran, Tehran, Iran.
Vet Med Sci. 2021 Mar;7(2):362-369. doi: 10.1002/vms3.368. Epub 2020 Sep 24.
Zoonotic Cutaneous Leishmaniasis is increasing in the world and Phlebotomus papatasi as a proven vector was considered in different aspects for disease control. Sandfly saliva contains proteins which provoke host immune system. These proteins are candidates for developing vaccines.
The main purpose of this research was comparing evaluation of salivary glands proteomes from wild P. papatasi. Extracting these proteins and purifying of original SP15 as inducer agent in vector salivary glands from endemic leishmaniasis foci were other objectives.
Adult sandflies were sampled using aspirators and funnel traps from three endemic foci in 2017-2018. Each pair of salivary glands of unfed females was dissected and proteins were extracted using thermal shocking and sonication methods. Purification was performed through RP-HPLC. All equivalent fractions were added together in order to reach sufficient protein concentration. Protein content and profile determination were examined with SDS-PAGE.
The protein concentration of whole-salivary glands of specimens was determined approximately 1.6 µg/µl (Isfahan) and 1 µg/µl (Varamin and Kashan). SDS-PAGE revealed 10 distinct bands between 10 and 63 kDa. Analysis of proteomes showed some similarities and differences in the chromatograms of different foci. SDS-PAGE of all collected fractions revealed SP15-like proteins were isolated in 24 min from Varamin, 26 to 30 min from Kashan and 29.4 min from Isfahan and were around 15 kDa.
Isolation of salivary components of Iranian wild P. papatasi is very important for finding potential proteins in vaccine development and measuring control strategy of zoonotic cutaneous leishmaniasis in Iran and this could be concluded elsewhere in the world.
动物源皮肤利什曼病在全球范围内呈上升趋势，已证实的媒介白蛉（Phlebotomus papatasi）在疾病控制的各个方面都被考虑在内。白蛉唾液中含有能刺激宿主免疫系统的蛋白质。这些蛋白质是开发疫苗的候选物。
本研究的主要目的是比较来自野生白蛉（P. papatasi）的唾液腺蛋白质组的评估。提取这些蛋白质，并从利什曼病流行地区的媒介白蛉唾液腺中纯化原始 SP15 作为诱导剂是其他目的。
2017 年至 2018 年，使用吸气器和漏斗陷阱从三个流行地区采集成年沙蝇。每个未喂食雌性的一对唾液腺被解剖，并使用热休克和超声处理方法提取蛋白质。通过 RP-HPLC 进行纯化。为了达到足够的蛋白质浓度，将所有等效的馏分合并在一起。使用 SDS-PAGE 检查蛋白质含量和图谱的测定。
标本全唾液腺的蛋白质浓度约为 1.6 µg/µl（伊斯法罕）和 1 µg/µl（瓦拉明和卡尚）。SDS-PAGE 显示 10 个不同的条带在 10 到 63 kDa 之间。对蛋白质组的分析表明，不同焦点的色谱图存在一些相似和差异。对所有收集的馏分进行 SDS-PAGE 分析显示，来自瓦拉明的 SP15 样蛋白在 24 分钟内被分离出来，来自卡尚的在 26 到 30 分钟内，来自伊斯法罕的在 29.4 分钟内，大小约为 15 kDa。
分离伊朗野生白蛉的唾液成分对于寻找疫苗开发中的潜在蛋白质以及衡量伊朗动物源皮肤利什曼病的控制策略非常重要，这可以在世界其他地方得出结论。
