Abstract

KING-BATSIOS, Emmanuel N. et al. Mycobacterium tuberculosis recombinant protein Rv2626c expressed in Streptomyces lividans. Physico-chemical and Immunological characterization as potential vaccine antigen. Vaccimonitor [online]. 2019, vol.28, n.2, pp. 68-79.  Epub Aug 01, 2019. ISSN 1025-0298.

Mycobacterium tuberculosis (Mtb) is a leading cause of death globally. Latent tuberculosis infection threatens 1.7 billion people. Mtb latency is mediated by a group of proteins, mainly coded by the Dormancy Safety Regulator (DosR). The protein Rv2626c is the strongest regulated member of this operon. Previous results, including ours, indicate a strong potential of Rv2626c as antigen in a new multiple tuberculosis vaccine. Objectives of this study were to purify the rRv2626c protein and characterize it physico-chemically and immunologically. The purified protein migrates as a sole band after a non-reductive PAGE-silver staining. Under reductive conditions, the dimer isoform appearing at 30.9 kDa prevails over the monomer 15.6 kDa. Mass spectrometry corroborates electrophoresis results regarding dimer molecular weight, of approximately 32 kDa. Six of its digested peptides matched those of HRP-1 protein (Rv2626c) of Mtb whereas 92.1% of its amino acid sequence contains three mutations and the addition of an amino acid. With respect to native Mtb protein, 12 of the 13 main epitopes are conserved. Antigenicity was corroborated in volunteers, the antibody responses were significantly higher in a number of infected tuberculosis patients in comparison to healthy Mantoux negative donors as well as in mice immunized with reference Rv2626c, while the immune identification pattern was as expected. The purified protein was able to elicit strong immune response in mice and the resulting antibodies recognized the reference Rv2626c protein. Lastly, the productive specific yield of the Streptomyces lividans strain is sustainable. Taking these results altogether, corroborates our rRv2626c as a promising candidate as antigen for new tuberculosis vaccine formulations.

Keywords : Mycobacterium tuberculosis; Rv2626c recombinant protein; Streptomyces lividans; tuberculosis vaccine.

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