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Background   Arcanobacterium haemolyticum  is an emerging human pathogen that causes pharyngitis, wound infections, and a variety of occasional invasive diseases. Since its initial discovery in 1946, this Gram positive organism has been known to have hemolytic activity, yet no hemolysin has been previously reported.  A. haemolyticum  also displays variable hemolytic activity on laboratory blood agar that is dependent upon which species the blood is derived.    Results  Here we describe a cholesterol-dependent cytolysin (CDC) secreted by  A. haemolyticum , designated arcanolysin ( aln ), which is present in all strains (n = 52) tested by DNA dot hybridization. Among the known CDCs, ALN is most closely related to pyolysin (PLO) from  Trueperella  (formerly  Arcanobacterium )  pyogenes . The  aln  probe, however, did not hybridize to DNA from  T. pyogenes . The  aln  open reading frame has a lower mol %G+C (46.7%) than the rest of the  A. haemolyticum  genome (53.1%) and is flanked by two tRNA genes, consistent with probable acquisition by horizontal transfer. The ALN protein (~ 64 kDa) contains a predicted signal sequence, a putative PEST sequence, and a variant undecapeptide within domain 4, which is typically important for function of the toxins. The gene encoding ALN was cloned and expressed in  Escherichia coli  as a functional recombinant toxin. Recombinant ALN had hemolytic activity on erythrocytes and cytolytic activity on cultured cells from human, rabbit, pig and horse origins but was poorly active on ovine, bovine, murine, and canine cells. ALN was less sensitive to inhibition by free cholesterol than perfringolysin O, consistent with the presence of the variant undecapeptide.    Conclusions  ALN is a newly identified CDC with hemolytic activity and unique properties in the CDC family and may be a virulence determinant for  A. haemolyticum .

Arcanolysin is a cholesterol-dependent cytolysin of the human pathogen Arcanobacterium haemolyticum