Shirazian Maryam, Taghipour Niloofar, Akhavan Amir Ahmad, Tabaei Seyyed Javad Seyyed, Abaei Mohammad Reza, Firouzjaie Fahimeh, Fatemi Mahboubeh, Mosaffa Nariman, Moin Vaziri Vahideh
Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Exp Parasitol. 2024 Dec;267:108863. doi: 10.1016/j.exppara.2024.108863. Epub 2024 Nov 23.
Early interactions between Leishmania-macrophages (MQ) of host and effect of sand fly saliva are central to leishmaniasis outcome. Macrophages are able to kill or act as long-term hosts of parasite depending on host immunity. It proved that immunogenic proteins in sand fly saliva mostly have an exacerbating effect on leishmaniasis by up-regulating cytokines. We have explored expression of Arginase 1 (ARG1) and inducible Nitric Oxide Synthase (iNOS) genes in macrophages of three different rodents, BALB/c (susceptible), C57BL/6 (resistant) and Rhombomys opimus (R. opimus, natural reservoir) in presence of Leishmania major (L. major), salivary gland homogenate (SGH) of Phlebotomus papatasi (Ph. papatasi) and finally L. major + SGH. Stationary phase of promastigotes was used; salivary glands were extracted from female of Ph. papatasi (3-5 day-old/non-blood fed). SGH prepared by sonication. Macrophages harvested from the peritoneal cavity of each rodent and grouped as follow; 1) macrophage (control group), 2) MQ + L.major 3) MQ + SGH 4) MQ + L.major + SGH. After 6 h of incubation, culture medium supernatant collected, RNA extraction and cDNA synthesis performed, expression level of desired genes checked by Real-time PCR. ARG1 expression pattern in MQ + SGH and MQ + L.major group showed the highest and lowest expressions level respectively in BALB/c and C57BL/6. But, in MQ + L.major + SGH, although the highest ARG1 expression happened in BALB/c again, the lowest one observed in R. opimus. On the other hand, iNOS expression showed significant increase in all treated group of C57BL/6 macrophages. Interestingly, iNOS expression showed significant differences in MQ + L.major group of C57BL/6 in comparison to other rodents. Expression of ARG1 and iNOS in macrophages of BALB/c and C57BL/6 and R. opimus are different and can justify their clinical outcome of disease. The difference in gene expression pattern is related to the genetics of host and shows that genetic differences between hosts can affect the immune responses caused by saliva proteins even of the same species of sand fly.
宿主的利什曼原虫与巨噬细胞（MQ）之间的早期相互作用以及白蛉唾液的影响是利什曼病结局的关键因素。巨噬细胞能够根据宿主免疫力杀死寄生虫或成为其长期宿主。事实证明，白蛉唾液中的免疫原性蛋白大多通过上调细胞因子对利什曼病起加重作用。我们研究了在存在硕大利什曼原虫（L. major）、巴氏白蛉（Ph. papatasi）唾液腺匀浆（SGH）以及最终的L. major + SGH的情况下，三种不同啮齿动物（BALB/c（易感）、C57BL/6（抗性）和大沙鼠（R. opimus，自然宿主））巨噬细胞中精氨酸酶1（ARG1）和诱导型一氧化氮合酶（iNOS）基因的表达。使用前鞭毛体的稳定期；从3至5日龄未吸血的雌性巴氏白蛉中提取唾液腺。通过超声处理制备SGH。从每只啮齿动物的腹腔中收获巨噬细胞并分组如下：1）巨噬细胞（对照组），2）MQ + L. major，3）MQ + SGH，4）MQ + L. major + SGH。孵育6小时后，收集培养基上清液，进行RNA提取和cDNA合成，通过实时PCR检测所需基因的表达水平。在BALB/c和C57BL/6中，MQ + SGH组和MQ + L. major组的ARG1表达模式分别显示出最高和最低表达水平。但是，在MQ + L. major + SGH中，尽管ARG1的最高表达再次出现在BALB/c中，但最低表达出现在大沙鼠中。另一方面，C57BL/6巨噬细胞的所有处理组中iNOS表达均显著增加。有趣的是，与其他啮齿动物相比，C57BL/6的MQ + L. major组中iNOS表达存在显著差异。BALB/c、C57BL/6和大沙鼠巨噬细胞中ARG1和iNOS的表达不同，这可以解释它们疾病的临床结局。基因表达模式的差异与宿主的遗传学有关，表明宿主之间的遗传差异会影响由同一种白蛉唾液蛋白引起的免疫反应。
