IgG Endopeptidase SeMac does not Inhibit Opsonophagocytosis of Streptococcus equi Subspecies equi by Horse Polymorphonuclear Leukocytes

Abstract

The secreted Mac protein made by group A Streptococcus (GAS) inhibits opsonophagocytosis of GAS by human polymorphonuclear leukocytes (PMNs). This protein also has the endopeptidase activity against human immunoglobulin G (IgG), and the Cys94, His262 and Asp284 are critical for the enzymatic activity. The horse pathogen Streptococcus equi subspecies equi produces a homologue of Mac (SeMac). SeMac was characterized to determine whether SeMac has IgG endopeptidase activity and inhibits opsonophagocytosis of S. equi by horse PMNs. The gene was cloned and recombinant SeMac was overexpressed in Escherichia coli and purified to homogeneity. Mice with experimental S. equi infection and horses with strangles caused by S. equi seroconverted to SeMac, indicating that SeMac is produced in vivo during infection. SeMac has endopeptidase activity against human IgG. However, the protein just cleaves a small fraction, which may be IgG1 only, of horse IgG. Replacement of Cys102 with Ser or His272 with Ala abolishes the enzymatic activity of SeMac, and the Asp294Ala mutation greatly decreases the enzymatic activity. SeMac does not inhibit opsonophagocytosis of S. equi by horse PMNs but opsonophagocytosis of GAS by human PMNs. Thus, SeMac is a cysteine endopeptidase with a limited activity against horse IgG and must have other function.

Streptococcus equi: a pathogen restricted to one host

“…These enzymes cleave IgG produced by a variety of animal species, thereby reducing IgG recognition and targeting of bacteria by the immune response. IdeE2 and IdeZ2 cleave equine IgG more efficiently than IdeE and IdeZ (Hulting et al, 2009), but there remains some uncertainty as to whether the main antiphagocytic mechanism exploited by IdeE is reduction of opsonophagocytosis via IgG cleavage (Liu & Lei, 2010;. Rather, its dose dependent inhibitory effect on the bactericidal activity of neutrophils for both S. equi and S. zooepidemicus, which is neutralized by a specific antibody response, suggests a different mechanism of reaction.…”