Biotechnology and Bioprocess Engineering 2019; 24(5): 773-781  
Chimeric Myostatin - Tetanic Toxin Epitopes and Heterologous Prime-boost Immunization Improve Immune Response Stimulating Muscle Growth in Mice
Vianey Ramírez Andoney1, Amanda Gayosso Vázquez1, Juan Pablo Pintor Ríos1, Jorge Enrique Vázquez Buchelli1, and Rogelio A. Alonso Morales1,2,*
1Department of Genetic and Biostatistics, Faculty of Veterinary Medicine and Zootechnics, National Autonomous University of Mexico, Mexico City 04510, Mexico
2Molecular Genetics Laboratory, Department of Genetic and Biostatistics, Faculty of Veterinary Medicine and Zootechnics, National Autonomous University of Mexico, Mexico City 04510, Mexico
Correspondence to: Rogelio A. Alonso Morales
Molecular Genetics Laboratory, Department of Genetic and Biostatistics, Faculty of Veterinary Medicine and Zootechnics, National Autonomous University of Mexico, Mexico City 04510, Mexico
Tel: +52-55-56225956
E-mail: ralonsom@unam.mx
Received: March 17, 2019; Revised: June 16, 2019; Accepted: June 17, 2019; Published online: October 31, 2019.
© The Korean Society for Biotechnology and Bioengineering. All rights reserved.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Myostatin is a transforming growth factor-β family member who acts as a negative regulator of skeletal muscle growth. The interference of its biological activity could increase skeletal muscle growth with clinical and animal production applications. A strategy to block the myostatin action is by the induction of an immune response against it. In this work, we evaluated as an immunogen a recombinant myostatin fused to the tetanic toxin T- helper epitopes P2 and P30. Genetic constructs of the chimeric myostatin were cloned in an expression vector and used as a DNA vaccine. Besides, a chimeric genetic construct, P2-miostatin–P30 was expressed in Escherichia coli, obtaining a recombinant chimeric antigen. To find out the functionality of these genetic constructs as a vaccine in inducing muscle growth responses, experimental groups of BALB/c mice were DNA immunized with the myostatin fused to P2, P30 or both. Furthermore, to improve the immune response, a heterologous prime–boost immunization scheme was evaluated where the DNA inoculation was followed by immunization with the recombinant antigen P2-myostatin-P30. The different body segments weight was recorded in control and vaccinated mice groups, finding increased muscle masses in the vaccinated groups. These experiments showed the effectiveness of the P2 and P30 T-helper epitopes in inducing an immune response to the fused myostatin, leading to muscle growth. The heterologous prime-boost immunization protocol is a promising vaccination strategy reducing the time and amount of antigen used to induce a immune response to myostatin.
Keywords: myostatin, prime boost immunization, tetanic toxin epitopes, DNA vaccine


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