PDF(Pubmed)
Abstract：
Mammalian models, such as murine, are used widely in pathophysiological studies because they have a high degree of similarity in body temperature, metabolism, and immune response with humans. However, non-vertebrate animal models have emerged as alternative models to study the host-pathogen interaction with minimal ethical concerns. Galleria mellonella is an alternative model that has proved useful in studying the interaction of the host with either bacteria or fungi, performing drug testing, and assessing the immunological response to different microorganisms. The G. mellonella immune response includes cellular and humoral components with structural and functional similarities to the immune effectors found in higher vertebrates, such as humans. An important humoral effector stimulated during infections is apolipophorin III (apoLp-III), an opsonin characterized by its lipid and carbohydrate-binding properties that participate in lipid transport, as well as immunomodulatory activity. Despite some parameters, such as the measurement of phenoloxidase activity, melanin production, hemocytes counting, and expression of antimicrobial peptides genes are already used to assess the G. mellonella immune response to pathogens with different virulence degrees, the apoLp-III quantification remains to be a parameter to assess the immune response in this invertebrate. Here, we propose an immunological tool based on an enzyme-linked immunosorbent assay that allows apoLp-III quantification in the hemolymph of larvae challenged with pathogenic agents. We tested the system with hemolymph coming from larvae infected with Escherichia coli, Candida albicans, Sporothrix schenckii, Sporothrix globosa, and Sporothrix brasiliensis. The results revealed significantly higher concentrations of apoLp-III when each microbial species was inoculated, in comparison with untouched larvae, or inoculated with phosphate-buffered saline. We also demonstrated that the apoLp-III levels correlated with the strains\' virulence, which was already reported. To our knowledge, this is one of the first attempts to quantify apoLp-III, using a quick and easy-to-use serological technique.
摘要：
哺乳动物模型,比如鼠类，在病理生理学研究中广泛使用，因为它们在体温方面具有高度的相似性，新陈代谢，和人类的免疫反应。然而,非脊椎动物动物模型已成为研究宿主-病原体相互作用的替代模型，而伦理问题最少。Galleriamellonella是一种替代模型，已被证明可用于研究宿主与细菌或真菌的相互作用。进行药物测试，并评估对不同微生物的免疫反应。G.mellonella免疫反应包括细胞和体液成分，其结构和功能与高等脊椎动物中的免疫效应物相似。比如人类。在感染过程中刺激的重要体液效应是载脂蛋白III（apoLp-III），一种调理素，其特征是参与脂质运输的脂质和碳水化合物结合特性，以及免疫调节活性。尽管有一些参数，如酚氧化酶活性的测量，黑色素的产生，血细胞计数,和抗菌肽基因的表达已经被用于评估不同毒力程度的病原体的免疫应答。apoLp-III定量仍然是评估该无脊椎动物免疫反应的参数。这里,我们提出了一种基于酶联免疫吸附测定的免疫学工具，该工具可以对被病原体攻击的幼虫的血淋巴进行apoLp-III定量。我们用感染了大肠杆菌的幼虫的血淋巴对系统进行了测试，白色念珠菌,申克氏孢子虫,球形孢子虫，和巴西孢子丝菌。结果表明，当接种每种微生物物种时，apoLp-III的浓度显着升高，与未接触的幼虫相比，或用磷酸盐缓冲盐水接种。我们还证明了apoLp-III水平与菌株的毒力相关，这已经被报道了。据我们所知,这是量化apoLp-III的首次尝试之一，使用快速和易于使用的血清学技术。
