Evaluation of neutralization activity of anti-tetanus monoclonal antibodies in cellular assays

• Grant number: 22/00205-0
• Support Opportunities: Scholarships in Brazil - Master
• Start date: March 01, 2022
• End date: February 29, 2024
• Field of knowledge: Biological Sciences - Immunology - Applied Immunology
• Principal Investigator: Ana Maria Moro
• Grantee: Cauã Pacheco de Lima
• Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
• Associated research grant: 20/07040-1 - Center of Research and Development in Immunobiologicals, AP.NPOP

Abstract

Tetanus remains a global health issue and reflects the precariousness of the health public system in underdeveloping countries. Although the complete immunization schedule is offered in Brazil, the Ministry of Health of Brazil disclose about a hundred new cases of accidental tetanus by year. In cases of infection, it is recommended the use of animal or human serum (passive immunization), in association with antibiotics. The alternative of anti-tetanus serum by human monoclonal antibodies (mAbs) is promising and represents an option more efficient and safer. In vitro assays assessing the effect of anti-tetanus mAbs incite future prophylaxis and new understanding about the disease pathogenesis. In this project, it is proposed the in vitro evaluation of the effect of anti-tetanus antibodies (mAbs) developed at the Laboratory of Biopharmaceuticals. The proposal is to use cellular assays to evaluate the interaction between toxins and neurons and its inhibition by mAbs. Neuro-2a cells (or other neuronal lineages) will be incubated with: (i) the tetanus toxin; (ii) tetanus toxin C-fragment and (iii) recombinant toxin catalytically inactive mutant (R372A/Y375F). The neutralization effect will be determined by mapping endocytic proteins, detecting molecular signals in anti-apoptotic survival pathways, and immunophenotyping analyses. We expect that the neutralization by individual or combined mAbs provides results in consonance with the in vivo assays previously performed and can be useful to evaluate the available panel of anti-tetanus human mAbs. (AU)