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Immunization and chemical conjugation of Bm95 obtained from Pichia pastoris enhances the immune response against vaccinal protein and Neisseria meningitidis capsular polysaccharide

Authors Rodriguez-Valle M, Canan-Hadden L, Niebla O

Received 4 November 2013

Accepted for publication 16 January 2014

Published 7 March 2014 Volume 2014:4 Pages 15—22


Checked for plagiarism Yes

Review by Single-blind

Peer reviewer comments 5

Manuel Rodriguez-Valle,1 Leonardo Canan-Hadden,2 Olivia Niebla2

1Animal Biotechnology Division, 2Analytical Division, Centre for Genetic Engineering and Biotechnology, Havana, Cuba

Abstract: The ectoparasite Rhipicephalus (Boophilus) microplus causes severe economic losses to the cattle industry in tropical and subtropical regions, and transmits endoparasites, such as Babesia bovis. The glycoprotein Bm95 is homologous to Bm86, a surface membrane protein of gut epithelial cells in R. microplus, and has been shown to efficiently control this ectoparasite in regions of the Americas. The immunostimulant properties of Bm86 have already been demonstrated after its coinjection with hepatitis B surface antigen (HBsAg) and the infectious bovine rhinotracheitis virus. This study evaluated the carrier and immunostimulant properties of Bm95 using low immunogenic Neisseria meningitidis capsular C polysaccharide (Men CpS) and HBsAg. We produced two polysaccharide-Bm95 conjugates by carbodiimide (MenCpSBm-c) and reductive amination (MenCpSBm-ra) methods. These conjugates were characterized and evaluated in mice. Antibody titers against Men CpS were significantly higher in mice immunized with MenCpSBm-ra (2,350±250, P<0.01) than in those immunized with MenCpSBm-c (250±75) or Men CpS (570±104). The study data indicate effective immunological memory after booster inoculation in mice immunized with MenCpSBm-ra. Additionally, significant humoral immunity against HBsAg was documented in mice coimmunized via the intranasal route with recombinant Bm95 (11,400±345) and HBsAg (128,000±250) compared with mice immunized only with HBsAg (400±40) or Bm95 (5,461±150, P<0.01). In conclusion, the immunostimulatory properties of recombinant Bm95 make it a useful element for developing safer conjugated vaccines against bacterial pathogens and for evaluation against ticks and tick-borne diseases in the context of a polyvalent veterinary vaccine.

Keywords: glycoconjugate, Bm86, capsular polysaccharides, carrier protein

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