Abstract
TRAIL, tumor necrosis factor (TNF)-related apoptosis-inducing ligand belongs to one of important cytokine superfamilIES, tumor necrosis factor (TNFα) . TRAIL-2 receptor agonists activate several cell signaling pathways in cells in different manners and could lead to apoptosis or necrosis. Agonistic egg yolk antibodies like IgY which have been developed in a selective manner could activate TRAIL death receptors such as TRAIL-2 (DR5) and thus apoptosis signaling. We here investigated induction of apoptosis in human breast cancer cells (MCF7 cell line) by an IgY produced against an 21 aminoacid epitope of the human TRAIL-2 receptor. As the first step a small peptide of 21 aminoacids choosen from the extracellular domain of DR5 protein was produced with a peptide synthesizer. After control assays and confirmation of the correct amino acid sequence, it was injected to hens immunized to achieve high affinity IgYs. At the next step, the produced IgYs were extracted and examined for specificity against DR5 protein by ELISA assay. Subsequently, the anticancer effect of such IgYs was determined by MTT assay in the MCF7 human breast cancer cell line. The produced peptides successfully immunized hens and the produced antibodies which accumulated in egg yolk specifically recognized the DR5 protein. IgYs exerted significant toxicity and killed MCF7 cells as shown by MTT assay.
Highlights
Cancer is a disease of a genome and normality of cell proliferation
Agonistic egg yolk antibodies like IgY which have been developed in a selective manner could activate TRAIL death receptors such as TRAIL-2 (DR5) and apoptosis signaling
Efficient production of IgY against little epitope of DR5 To test the production of antibody after immunization period, we performed the ELISA assay with IgY samples
Summary
Cancer is a disease of a genome and normality of cell proliferation. In these types of cells, several cycles of duplication without right replication cause amplification of mutations. Occupinig the death receptors from TNFα superfamilly such as CD95 or TRAIL receptors by their specific ligands can recruit the subsequent effector molecules like caspase 8 which will lead to performing complex death inducing signal complex (DISC) (Ashkenazi, 20008). It will activate, or alternatively permeablize mitochondrial membrane by cleaving Bid and releasing the cytochrome c out of it. At least, both of these two pathways cause apoptosis in cells. TRAIL, selectively induces apoptosis in cancerous cells by their specialized death inducing receptors
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