title: Identification of anti-alpha toxin monoclonal antibodies that reduce the severity of Staphylococcus aureus dermonecrosis and exhibit a correlation between affinity and potency.
abstract: Staphylococcus aureus alpha toxin (AT) is an important virulence determinant and may be a valid target for immunoprophylaxis against staphylococcal disease. Here we report the identification of potent inhibitory anti-AT monoclonal antibodies (MAbs) derived using B-cell hybridoma technology from VelocImmune mice engineered to produce IgG with a human variable domain. A small panel of inhibitory MAbs blocked AT-mediated lysis of rabbit red blood cells, A549 human lung epithelial cells, and THP-1 human monocytic cells, in a dose-dependent manner. Binding studies indicated that these MAbs recognize a similar epitope on AT and exhibit dissociation constants (K(D)) ranging from 0.50 to 15 nM. In an S. aureus dermonecrosis model, mice passively immunized with anti-AT inhibitory MAbs exhibited significant reductions of lesion size relative to mice treated with an irrelevant IgG control. Interestingly, there was a correlation between MAb affinity for a single epitope, the 50% inhibitory concentration (IC(50)) in the AT hemolytic assay, and lesion size reduction in the dermonecrosis model. A representative high-affinity MAb, 2A3.1, was demonstrated to significantly reduce lesion size following infection with three different clinical isolates (USA300, CC30, and CC5). Taken together, these results indicate that in vitro potency of anti-AT MAbs predicts in vivo potency in this model, supporting their continued preclinical evaluation as molecules for immunoprophylaxis against staphylococcal skin and soft tissue infections caused by diverse clinical isolates.
authors.affiliation: Department of Infectious Disease, MedImmune, LLC, Gaithersburg, Maryland, USA.
authors.forename: C
authors.initials: C
authors.lastname: Tkaczyk
authors.forename: L
authors.initials: L
authors.lastname: Hua
authors.forename: R
authors.initials: R
authors.lastname: Varkey
authors.forename: Y
authors.initials: Y
authors.lastname: Shi
authors.forename: L
authors.initials: L
authors.lastname: Dettinger
authors.forename: R
authors.initials: R
authors.lastname: Woods
authors.forename: A
authors.initials: A
authors.lastname: Barnes
authors.forename: R S
authors.initials: RS
authors.lastname: MacGill
authors.forename: S
authors.initials: S
authors.lastname: Wilson
authors.forename: P
authors.initials: P
authors.lastname: Chowdhury
authors.forename: C K
authors.initials: CK
authors.lastname: Stover
authors.forename: B R
authors.initials: BR
authors.lastname: Sellman
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage* / Q000494:pharmacology; D000992:Antitoxins / Q000008:administration & dosage* / Q000494:pharmacology; D001427:Bacterial Toxins / Q000037:antagonists & inhibitors* / Q000633:toxicity*; D002460:Cell Line; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004306:Dose-Response Relationship, Immunologic; D004847:Epithelial Cells / Q000187:drug effects; D004912:Erythrocytes / Q000187:drug effects; D005260:Female; D006460:Hemolysin Proteins / Q000037:antagonists & inhibitors* / Q000633:toxicity*; D006801:Humans; D007116:Immunization, Passive; D020128:Inhibitory Concentration 50; D051379:Mice; D008807:Mice, Inbred BALB C; D009000:Monocytes / Q000187:drug effects; D009336:Necrosis / Q000473:pathology / Q000517:prevention & control*; D011817:Rabbits; D013207:Staphylococcal Skin Infections / Q000473:pathology / Q000517:prevention & control*
chemical_list: D000911:Antibodies, Monoclonal; D000992:Antitoxins; D001427:Bacterial Toxins; D006460:Hemolysin Proteins; C021091:staphylococcal alpha-toxin