Surface-Expressed Mig Protein Protects Streptococcus dysgalactiae against Phagocytosis by Bovine Neutrophils

Abstract

ABSTRACT The mig gene of Streptococcus dysgalactiae , a major bovine mastitis pathogen, encodes two plasma protein-binding receptors, α 2 -macroglobulin (α 2 -M) and immunoglobulin G (IgG). In this study, the mig gene from one S. dysgalactiae isolate was cloned and expressed in Escherichia coli . The IgG receptor region encoded by mig was conserved in 16 S. dysgalactiae strains. An isogenic mig mutant was constructed by allele replacement mutagenesis of the wild-type gene in S. dysgalactiae . The IgG-binding activity was lost in the mig mutant strain, whereas the α 2 -M receptor activity was still expressed but was detected only in the culture supernatant. In flow cytometry phagocytosis and bacterial-colony-counting bactericidal assays, the wild-type strain was found to be significantly more resistant to phagocytosis and killing by bovine neutrophils (PMNs) than the mig mutant strain when bacteria were preincubated with bovine serum. We therefore speculate that the Mig protein of S. dysgalactiae plays a role in virulence of the bacteria by binding to the plasma protein α 2 -M or IgG and thus preventing phagocytosis by bovine PMNs.