Apoptotic Killing of Breast Cancer Cells by IgYs Produced Against a Small 21 Aminoacid Epitope of the Human TRAIL-2 Receptor

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.

Egg yolk antibodies (IgY) and their applications in human and veterinary health: A review

“…Following this rationale, Amirijavidv et al produced highly specific IgY against a sequence of 21 amino acids present on the ectodomain of the TRAIL (TNF-related apoptosis-inducing ligand) receptor TRAIL-R2 (DR5). The antibodies bound to the amino acid sequence and activated the DR5 receptors in human breast cancer cells MCF7, acting as a TRAIL agonist and inducing apoptosis [87]. IgY against other receptors, such as the HER2 receptor, was tested coupled to single walled carbon nanotubes (SWNTs) and specifically detected the HER2 receptors on the surface of SK-BR-3 cells.…”

IgY Antibodies as Biotherapeutics in Biomedicine

“…Information related to the pharmacokinetics of IgY is currently scarce. The available data come from animal models with the application of sublethal doses of xenogeneic snake venom [ 60 ]. In these studies, IgY was shown to have a wide and rapid distribution in the animal body, including muscles and brain parenchyma [ 61 ].…”

“…Similarly, the entry of IgY into the brain across the blood–brain barrier was demonstrated, and it could neutralize the poison and act as an antivenom. The IgY antibodies specifically recognized the venom and as previously mentioned, did not activate the complement system, suggesting the possibility of safe re-administration until the resolution of intoxication symptoms in animals not developing ADE [ 60 , 61 ]. Since oral administration of IgY in humans has been shown to be effective and without side-effects, its parenteral administration needs further investigation to determine its safety, tolerability, and efficacy [ 62 , 63 , 64 ].…”

Bioactive Minor Egg Components

“…This approach using chicken igy has shown promising results in killing breast cancer cells. 121,122 14.3.3.1.3 Egg Proteins and Diagnosis of Cancer. in parallel, the immunoglobulin y strategy to produce antibodies against cancerous biomarkers has been extensively used to develop tools (eliSA/immunohistochemistry) that contribute to the diagnosis/prognosis of the disease.…”