Sampaio Luzia Cristina Lencioni, Baldissera Matheus Dellaméa, Grando Thirssa Helena, Gressler Lucas Trevisan, Capeleto Dianni de Menezes, de Sa Mariângela Facco, de Jesus Francielli Pantella Kuns, dos Santos Alceu Gonçalves, Anciuti Andreia Nobre, Colonetti Karina, Stainki Daniel Roulim, Monteiro Silvia Gonzalez
Department of Microbiology and Parasitology, Universidade Federal de Santa Maria (UFSM), Brazil.
Department of Microbiology and Parasitology, Universidade Federal de Santa Maria (UFSM), Brazil.
Vet Parasitol. 2014 Aug 29;204(3-4):96-103. doi: 10.1016/j.vetpar.2014.05.032. Epub 2014 May 28.
The use of avian antibodies has aroused interest in biomedical research due to the numerous advantages compared to mammal's antibodies. Our study aimed to produce and purify IgY immunoglobulins in order to use as an alternative therapy against Trypanosoma evansi. Every 14 days, four New Hampshire chickens were immunized with trypomastigotes of T. evansi, totaling five inoculations. Eggs were collected during 70 days and the extraction of IgY was performed by precipitation through the PEG-6000 method. Characterization and purification of IgY anti-T. evansi were carried out by SDS-PAGE and Western blot, where heavy and light chains were detected. The production of IgY was noted during the whole period, and the average production was 2.87 ± 0.14 at the end of this study. Sample's titration allowed the quantification of specific IgY anti-T. evansi, with antibodies produced showing high avidity indexes. The results indicated that T. evansi is able to generate an immune response in poultry, resulting in a production of specific antibodies. In vivo test showed that IgY treatment resulted in increase of prepatent period, longevity and survival of infected animals, when compared with the positive control, demonstrating an initial, but no curative, trypanocidal activity.
与哺乳动物抗体相比，禽类抗体具有诸多优势，这使得其在生物医学研究中备受关注。我们的研究旨在生产和纯化卵黄免疫球蛋白（IgY），以便用作对抗伊氏锥虫的替代疗法。每14天用伊氏锥虫的锥鞭毛体对4只新罕布什尔鸡进行免疫，共接种5次。在70天内收集鸡蛋，并通过聚乙二醇-6000（PEG-6000）沉淀法提取IgY。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和蛋白质免疫印迹法对伊氏锥虫特异性IgY进行表征和纯化，检测到了重链和轻链。在整个期间均观察到了IgY的产生，在本研究结束时平均产量为2.87±0.14。样品滴定可对伊氏锥虫特异性IgY进行定量，所产生的抗体显示出高亲和力指数。结果表明，伊氏锥虫能够在禽类中引发免疫反应，从而产生特异性抗体。体内试验表明，与阳性对照相比，IgY治疗可延长感染动物的潜伏期、寿命和生存期，显示出初步但非治愈性的杀锥虫活性。
