Disruption of the Putative Vascular Leak Peptide Sequence in the Stabilized Ricin Vaccine Candidate RTA1-3344-198.docVIP

Toxins 2013, 5, 224-248; doi:10.3390/toxins5020224 OPEN ACCESS toxins ISSN 2072-6651 /journal/toxins Article Disruption of the Putative Vascular Leak Peptide Sequence in the Stabilized Ricin Vaccine Candidate RTA1-33/44-198 Laszlo Janosi , Jaimee R. Compton , Patricia M. Legler *, Keith E. Steele , Jon M. Davis 4, 1 2 3, 1,? 1 Gary R. Matyas and Charles B. Millard 5 1 Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA; E-Mails: lacipaci@ (L.J.); steeleK@medI (K.E.S.); gmatyas@ (G.R.M.) 2 NOVA Research, Inc., Alexandria, VA 22308, USA; E-Mail: pton.ctr@ 3 Naval Research Laboratories, 4555 Overlook Ave., Washington, DC 20375, USA 4 United States Army Medical Research Institute of Infectious Diseases, Frederick, MD 21702, USA; E-Mail: jon.davis@ 5 U.S. Army Medical Research and Materiel Command, Fort Detrick, MD 21702-5012, USA; E-Mail: lard@ ? Current Address: MedImmune LLC, One Medimmune Way, Gaithersburg, MD 20878, USA * Author to whom correspondence should be addressed; E-Mail: patricia.legler@; Tel.: +1-202-404-6037; Fax: +1-202-404-8688. Received: 5 December 2012; in revised form: 10 January 2013 / Accepted: 11 January 2013 / Published: 29 January 2013 Abstract: Vitetta and colleagues identified and characterized a putative vascular leak peptide (VLP) consensus sequence in recombinant ricin toxin A-chain (RTA) that contributed to dose-limiting human toxicity when RTA was administered intravenously in large quantities during chemotherapy. We disrupted this potentially toxic site within the more stable RTA1-33/44-198 vaccine immunogen and determined the impact of these mutations on protein stability, structure and protective immunogenicity using an experimental intranasal ricin challenge model in BALB/c mice to determine if the mutations were compatible. Single amino acid substitutions at the positions corresponding with RTA D75 (to A, or N) and V76 (to