Abstract
Mycoplasma mycoides subspecies mycoides (Mmm) adhesion is tissue and host specific. Inhibition of adhesion will prevent Mmm from binding to lung cells and hence prevent colonization and disease. The aim of this study was to develop a panel of Mmm monoclonal antibodies against Mmm and use these antibodies to investigate their inhibitory effect on the adherence of Mmm to bovine lung epithelial cells (BoLEC), and to further identify an antigen to any of the inhibitory antibodies. Thirteen anti-Mycoplasma mycoides subsp. mycoides (AMMY) monoclonal antibodies (mAbs) inhibited adhesion by at least 30% and ten of the mAbs bound to multiple bands on Western blots suggesting that the antibodies bound to proteins of variable sizes. AMMY 10, a previously characterized Mmm- capsular polysaccharide (CPS) specific antibody, inhibited growth of Mmm in vitro and also caused agglutination of Mmm total cell lysate. AMMY 5, a 2-oxo acid dehydrogenase acyltransferase (Catalytic domain) (MSC_0267) specific antibody, was identified and polyclonal rabbit serum against recombinant MSC_0267 blocked adhesion of Mmm to BoLEC by 41%. Antigens recognized by these antibodies could be vaccine candidate(s) and should be subsequently tested for their ability to induce a protective immune response in vivo.
Highlights
Many mycoplasmas that infect livestock adhere to and colonize epithelial surfaces of various tissues in their hosts
The aim of this study was to identify potential Mycoplasma mycoides subspecies mycoides (Mmm) vaccine targets by developing a panel of monoclonal antibodies against Mmm, and use them to investigate their inhibitory effect on the adherence of Mmm to primary bovine lung epithelial cells (BoLEC), their capacity to inhibit Mmm growth in vitro, and to further identify an antigen to which any
The 13 antibodies were renamed anti-Mycoplasma mycoides subsp. mycoides (AMMY) 1–13 and their ability to inhibit adhesion of Mmm strain Afadé to BoLEC was tested at varying dilutions (Table 1) and with 3 different strains of Mmm (Fig. 1)
Summary
Many mycoplasmas that infect livestock adhere to and colonize epithelial surfaces of various tissues in their hosts. The cytoadherence mechanisms of many Mycoplasma species, including those of M. pneumoniae (Chaudhry et al, 2007), M. genitalium (Ueno et al, 2008), M. suis (Zhang et al., 2015), M. hominis (Brown et al, 2014), M. hyopneumonia (Burnett et al, 2006), M. agalactiae (Fleury et al, 2002) and M. bovis (Song et al, 2012) have been described. In these studies, adherence was shown to be a complex multifactorial process involving one or more adhesion molecules
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