Grando TH1, Baldissera MD2, de Sá MF2, do Carmo GM2, Porto BCZ2, Aguirre GSV2, Azevedo MI2, de Jesus FPK2, Santurio JM2, Sagrillo MR3, Stefani LM4, Monteiro SG5. 2017. J Immunol Methods. 449:56-61. doi: 10.1016/j.jim.2017.07.002. Epub 2017 Jul 8.
Trypanosoma cruzi is a flagellated protozoan belonging to the Trypanosomatidae family, the etiologic agent of Chagas disease. Currently, there is neither a licensed vaccine nor effective treatment, characterizing an unmet clinical need. The IgY refers to the egg yolk immunoglobulin (Y=yolk) and its production and use are subjects of many studies due to the diversity of its diagnostic and therapeutic applications. Several researchers have shown that the use of specific IgY may prevent and/or control infectious and parasitic diseases. Based on these evidences, the aim of this study was to immunize chickens with trypomastigotes of T. cruzi in order to produce highly effective and pure antibodies (IgY), as well as extract, characterize, quantify, and verify cytotoxic effects of IgY anti-T. cruzi. After the induction of IgY production by chickens, the eggs were collected and the IgY was extracted by method of precipitation of polyethylene glycol 6000. The IgY anti-T. cruzi characterization was performed using polyacrylamide gel electrophoresis (SDS-PAGE), western-blot and enzyme-linked immunosorbent assay (ELISA). Moreover, the cytotoxic or proliferative effects of IgY anti-T. cruzi was verified by MTT assay. The concentration of IgY in yolk was 8.41±1.47mg/mL. The characterization of IgY reveled bands of stained peptides with molecular weight between 75 and 50kDa and 37 and 25kDa. In the ELISA test was observed that there was antigen-antibody reaction throughout the sample period. The concentrations of 1, 5 and 10mg/mL of IgY anti-T. cruzi presented no cytotoxic of proliferative effects in mononuclear and VERO cells in vitro. The results indicated that T. cruzi is able to generate a high production of specific immunoglobulins in chickens, it did not cause damage to the cell membrane and no proliferative effect.
Avian immunoglobulins; Chagas disease; Cytotoxic; Egg yolk antibodies