Recombinant fusion protein of Heparin-Binding Hemagglutinin Adhesin and Fibronectin Attachment Protein (rHBHA-FAP) of Mycobacterium avium subsp. paratuberculosis elicits a strong gamma interferon response in peripheral blood mononuclear cell culture

Abstract

Background: Mycobacterium avium subsp. paratuberculosis (MAP) is a causative agent of Johne's disease in all ruminants worldwide. Economic problems in dairy cattle and sheep industries, public health concern, persistence of MAP in the environment and lack of effective vaccines mentioned necessity of research about various antigens to introduce as vaccine candidates. Based on MAP pathogenesis, it seems that research about the production of new recombinant proteins to stimulate cell-mediated immunity is helpful. This study describes successful expression and purification of a chimeric fusion protein which consists of Heparin-Binding Hemagglutinin Adhesin (HBHA) and high antigenic region of Fibronectin Attachment Protein (FAP-P). Triggered antigen-specific IFN-γ response of isolated PBMCs from immunized goats to rHBHA-FAP and all crude proteins of MAP (PPD), was measured by ELISA. Results: Significant increases were observed in the IFN-γ production level of peripheral blood mononuclear cells (PBMCs) stimulated by constructed chimeric protein from rHBHA-FAP and PPD vaccinated goats. Antigen-specific gamma interferon (IFN-γ) secretion in positive group (immunized by PPD) against rHBHA-FAP and test group (immunized by rHBHA-FAP) against PPD, also statistically insignificant rises between stimulation with rHBHA-FAP and PPD, suggested the potential and specificity of our chimeric protein to stimulate cell mediated immunity against MAP. Conclusions: Collectively, these results demonstrate that rHBHA-FAP elicits a strong IFN-γ production in PBMC culture. Therefore, further studies of the present product as a candidate vaccine in naturally infected animals should be conducted, to analyze its potential.

Immuno-reactivity evaluation of Mce-truncated subunit candidate vaccine against Mycobacterium avium subspecies paratuberculosis challenge in the goat models

“…After the failure of ‘test and cull’ policy, vaccination is one of the most cost effective control measures for this incurable infectious disease of livestock [ 5 ]. Various reports emphasized that vaccination is the best method for control of JD in animals [ 7 , 9 , 23 ]. However, the vaccines developed to date are not fully protective, they are not capable to prevent shedding of MAP in the feces and generate inadequate protection, serious granulomatous reaction and also interfere with tuberculin tests for Mycobacterium bovis [ 7 – 9 , 24 ].…”

Feature of Adhesins Produced by Human Clinical Isolates of Mycobacterium intracellulare, Mycobacterium intracellulare subsp. chimaera and Closely Related Species

“…One of the best characterized mycobacterial adhesins is the heparin-binding hemagglutinin (HBHA), initially identified in Mycobacterium tuberculosis and Mycobacterium bovis bacillus Calmette-Guérin (BCG) [ 17 , 18 ]. However, HBHA-like molecules are also present in many other mycobacteria, both pathogenic and nonpathogenic [ 19 , 20 , 21 , 22 ]. HBHA is located on the surface of the mycobacteria and mediates binding of the bacilli to epithelial cells and fibroblasts [ 18 ] by interacting with sulfated glycoconjugates present on the surfaces of host cells [ 23 ].…”

Socratic Lectures 8

“…There are a lot of research on engineered plasmids that can produce antigens in bacterial and mammalian hosts(Eraghi et al, 2017; Monreal-Escalante et al, 2021). Also, several purified proteins were investigated as vaccine candidates against MAP(Gupta et al, 2021;Eraghi et al, 2019;Fernández et al, 2022).…”