Detection of anticancer and apoptotic effect of the produced IgYs against the three extracellular domain of human DR5 protein

TNF alpha cytokine family in the body plays divers' roles in the cellular events such as cell proliferation, differentiation, necrosis, septic shock and apoptosis. In response to TNF therapy, several cell signaling pathways activated in cells which in different manners can lead to apoptosis or necrosis. However induction of apoptosis is depended on one of its important members, TRAIL and its receptors that will be followed by apoptosis activity. Tumor necrosis factor-related apoptosis-inducing ligand [TRAIL] and especially the DR5, is generating considerable interests as a possible anticancer therapeutic agent because of its selective activation in apoptosis of this receptor as a superior affinity to ligands. The study was performed in invitro assay and the anticancer effects of the produced antibodies were assumed by MTT and flowcytometric methods. In the first step for immunization of the hens, three selective small peptides from extracellular domain of DR5 which were chemically synthesized, injected to hens and after the proper immunization of them, IgYs were extracted from the egg yolk. After assumption of specificity of the purified IgYs against the whole DR5 protein, they were performed in MTT assay and flowcytometric colorimeter. After confirmation of synthesized peptides they were injected to hens with Fround`s complete adjuvant. With completing the immunization procedure the specificity of purified IgYs were confirmed by ELISA. The antibodies were significantly killed the MCF7 breast cancer cells, but had divers affect [proliferative] on normal hepatocyte cells. Additionally, significantly they induced apoptosis on the cancerous cells in contrast to control cells. The results clearly demonstrated that the produced IgYs with reduced cost and time managing could remarkably use as an effective anticancer drug

Anticancer effect of the IgY that produced against a small peptide with 15 amino acids of human DR5 on MCF7 cell line

Tumor necrosis factor- alpha [TNF-alpha] plays an important role in diverse cellular events such as septic shock, induction of other cytokines, cell proliferation and apoptosis. Tumor necrosis factor [TNF]-related apoptosis-inducing ligand [TRAIL] is currently attracting great interest as a potential anticancer drug. TRAIL could selectively induce apoptosis in tumor cells in vitro and in vivo by a death receptor-mediated process. TRAIL shows a high degree of promiscuity as it binds to the DR5 receptor and it is generating considerable interests as a possible anticancer therapeutic agent. Use of TRAIL or its antagonist could be a good anticancer treatment in future. The extracellular domain of DR5 human protein which has the attachment part of this ligand to TRAIL ligand is considerable domain of it. We produced a small peptide with just 15 aminoacids from this domain, with peptide synthesizer. Then inject them to hens to immunize them and achieve high affinity IgY. At least, obtained IgYs specially recognize DR5 protein and in vitro start exclusively to induce death in the MCF7 cell line, and interestingly not on normal cells