Muñoz F A, Franco-Noguez S Y, Gonzalez-Ballesteros E, Negrete-Philippe A C, Flores-Romo L
Facultad de Agronomía y Veterinaria, Universidad Autónoma de San Luis Potosí, Carretera San Luis-Matehuala km 14.5, Soledad de Graciano Sánchez, C.P.78433, San Luis Potosí, México,
Vet Res Commun. 2014 Jun;38(2):123-8. doi: 10.1007/s11259-014-9595-0. Epub 2014 Feb 26.
Phagocytosis is a fundamental aspect of innate immunity that is conserved across many species making it a potentially useful health-assessment tool for wildlife. In non-mammalian vertebrates, heterophils, monocytes, macrophages, melanomacrophages, and thrombocytes all have phagocytic properties. Recently, B lymphocytes from fish, amphibians, and aquatic turtles have also showed phagocytic capacity. Phagocytes can be studied by flow cytometry; however, the use of this tool is complicated in reptiles partly because nucleated erythrocytes complicate the procedure. We separated green turtle leukocytes by density gradient centrifugation and identified subpopulations by flow cytometry and confocal microscopy. Additionally, we assessed their ability to phagocytize Fluorspheres and Ovoalbumin-Alexa. We found that heterophils and lymphocytes but not monocytes could be easily identified by flow cytometry. While heterophils from adults and juvenile turtles were equally able to phagocytize fluorspheres, adults had significantly more phagocytic ability for OVA-Alexa. Lymphocytes had a mild phagocytic activity with fluorospheres (27-38 %; 39-45 %) and OVA-Alexa (35-46 %; 14-22 %) in juvenile and adult green turtles, respectively. Confocal microscopy confirmed phagocytosis of fluorospheres in both heterophils and lymphocytes. This provides the first evidence that green turtle lymphocytes have phagocytic activity and that this assay could potentially be useful to measure one aspect of innate immunity in this species.
吞噬作用是先天免疫的一个基本方面，在许多物种中都存在，这使其成为一种潜在有用的野生动物健康评估工具。在非哺乳动物脊椎动物中，嗜异性粒细胞、单核细胞、巨噬细胞、黑素巨噬细胞和血小板都具有吞噬特性。最近，鱼类、两栖类和水龟的B淋巴细胞也显示出吞噬能力。吞噬细胞可以通过流式细胞术进行研究；然而，在爬行动物中使用这种工具很复杂，部分原因是有核红细胞使操作变得复杂。我们通过密度梯度离心分离绿海龟白细胞，并通过流式细胞术和共聚焦显微镜鉴定亚群。此外，我们评估了它们吞噬荧光微球和卵清蛋白- Alexa的能力。我们发现，通过流式细胞术可以很容易地识别嗜异性粒细胞和淋巴细胞，而不是单核细胞。虽然成年海龟和幼年海龟的嗜异性粒细胞吞噬荧光微球的能力相同，但成年海龟对OVA - Alexa的吞噬能力明显更强。在幼年和成年绿海龟中，淋巴细胞对荧光微球（分别为27 - 38%；39 - 45%）和OVA - Alexa（分别为35 - 46%；14 - 22%）具有轻微的吞噬活性。共聚焦显微镜证实了嗜异性粒细胞和淋巴细胞中荧光微球的吞噬作用。这提供了首个证据，表明绿海龟淋巴细胞具有吞噬活性，并且该检测方法可能有助于测量该物种先天免疫的一个方面。
