Production and Evaluation of an Avian IgY Immunotoxin against CD133+ for Treatment of Carcinogenic Stem Cells in Malignant Glioma: IgY Immunotoxin for the Treatment of Glioblastoma

Abstract

Background. Glioblastoma is the most common malignant tumor of Central Nervous System. Despite the research in therapeutics, the prognosis is dismal. Malignant glioma stem cells (MGSCs) are a major cause of treatment failure and increasing tumor recurrence. In general, cancer stem cells (CSCs) express prominin-1 (CD133), considered as a potential therapeutic target. In this study, we produced an avian immunotoxin directed against the subpopulation of CD133+ CSCs within a malignant glioma. We used the avian IgY because it has various advantages as increased affinity to mammal antigens and inexpensive obtention of large amounts of specific antibodies (approximately 1 mg/per egg). The design, production, purification and use of IgY anti CD133 immunotoxin constitute an original goal of this research.Methods. The immunodominant peptide of CD133 was designed to immunize hens; also, the extracellular domain of CD133 was cloned to probe the IgY antibodies. In parallel, a recombinant abrin A chain was produced inE. coliin order to join it to the Fc domain of the anti-CD133 IgY to conform the immunotoxin. This anti-CD133 IgY anti-tumor immunotoxin was testedin vitroandin vivo. Results. The cytotoxicity of the immunotoxinin vitroshowed that IgY-abrin immunotoxin reduced 55% cell viability. After subcutaneous MGSCs implantation, the animals treated intraperitoneally or intratumorally with the IgY-abrin immunotoxin showed more than 50% decrease of tumor volume.Conclusion. Results showed that the IgY-abrin immunotoxin had cytotoxic activity against CD133+ MGSCs and provides a novel approach for the immunotherapy of glioblastoma.

Cancer Stem Cells: Significance in Origin, Pathogenesis and Treatment of Glioblastoma

“…Stemness markers also can be targeted by immune means, e.g., by cytotoxic avian IgY-immunotoxin against human CD133+ GSCs. In vitro, the IgY toxin was found to diminish cell viability by 55%, and in vivo it was associated with a decrease in tumor volume of more than 50% [ 112 ].…”

IgY Antibodies as Biotherapeutics in Biomedicine

“…IgY antibodies obtained from poultry and purified from egg yolk have been tested as tools for diagnosis and treatment of cancer, as well as infectious, autoimmune, and allergic diseases [ 5 , 6 , 18 , 25 , 26 , 27 ]. In particular, they have been used against viruses and bacteria which cause respiratory disorders in humans and animals [ 12 ].…”

“…Researchers from China, the United States, Canada, Japan, and Germany have generated the largest number of publications [ 3 ]. In addition, this technology could be implemented with relative ease in developing countries, such as Mexico, due to the high levels of bird production and extensive experience in their management, not only in the veterinary field but also in human medicine for the diagnosis and treatment of infectious diseases and cancer [ 4 , 5 , 6 ].…”

New Immunotherapeutic Approaches for Glioblastoma

“…This avian anti-CD133 IgY was fused with the A chain of the abrin toxin with the purpose to inactivate the ribosome. The anti-CD133-IgY-abrine showed to reduce cell viability of C6 CSC in vitro and decreased tumor volume after implantation of malignant glioma stem cells in vivo , representing a low-cost tool for the treatment of GBM [ 86 ]. Another target expressed in glioma stem cells is the Eph receptors, which are part of the receptor tyrosine kinase family.…”