Tularemia Vaccine Development Contract: Monthly Technical Report Period: 2/1/2010 to 2/28/2010 Due Date: 3/15/2010 and Prepared by: C. Rick Lyons, Barbara Griffith, Terry Wu, Bob Sherwood, Julie Wilder, Julie Hutt, Michelle Valderas, Trevor Brasel, Dana Pohlman, Mitch Magee, Kathryn Sykes, Stephen Johnston, Karl Klose, Bernard Arulanandam, Justin Skoble Contract No. ADB Contract No. HHSN266200500040-C N01-AI-50040 Contract Title: Tularemia Vaccine Development Team Performance Period: 2/1 to 2/28/2010 Contractor Name: University of New Mexico Health Science Center Contractor Address: Controller’s Office MSC-09-5220 1 University of New Mexico Albuquerque, New Mexico 87131-0001 Authors: C. Rick Lyons, MD, PhD and Barbara B. Griffith, MS Date of submission: 3/12/2010 Monthly Technical Report: Based on Contributions from ASU, Cerus/Aduro, LBERI, UNM and UTSA Page 1 of 6 Tularemia Vaccine Development Contract: Monthly Technical Report Period: 2/1/2010 to 2/28/2010 Due Date: 3/15/2010 and Prepared by: C. Rick Lyons, Barbara Griffith, Terry Wu, Bob Sherwood, Julie Wilder, Julie Hutt, Michelle Valderas, Trevor Brasel, Dana Pohlman, Mitch Magee, Kathryn Sykes, Stephen Johnston, Karl Klose, Bernard Arulanandam, Justin Skoble Contract No. HHSN266200500040-C ADB Contract No. N01-AI-50040 Section I: Purpose and Scope of Effort The Tularemia Vaccine Development Contract will lead to vaccine candidates, two animal models and cellular assays vital for testing vaccine efficacy. Sections II and III: Progress and Planning Presented by Institution 1. Active milestones: 2, 8, 9, 10(UNM/LBERI),11(UNM/LBERI), merged 12/13/14/15(UNM/LBERI), merged 17/18/19/20/21/22(UNM/LBERI), reopened 28, 29 (UNM/LBERI), 52, 53, 54, 55, 56, 57 2. Completed milestones: 1, 3, 4, 5, 6, 7, 16, 25, 26, 27, 32, 33, 34 (UNM/ASU), 35 (ASU/UNM), 36, 39, 40, 43 (UTSA), 48,49, 50, 51 3. Inactive milestones: 23, 24, 30, merged 37/38 (ASU J/ LBERI/ ASU S), 59 4. Milestones terminated after initiation: 41, 42, 44, 46, (MSCR will be written) 5. Milestones terminated before initiated: 43 (Cerus), 45, 47, 58, (MSCR will not be written) See the attached 5 Powerpoint presentations annotated with minutes 1. 2. 3. 4. 5. UTSA Monthly Technical Call 2/16/2010 ASU Monthly Technical Call 2/23/2010 LBERI Monthly Technical Call 3/2/2010 UNM Monthly Technical Call 3/3/2010 Cerus Monthly Technical Call 3/9/2010 Page 2 of 6 Tularemia Vaccine Development Contract: Monthly Technical Report Period: 2/1/2010 to 2/28/2010 Due Date: 3/15/2010 and Prepared by: C. Rick Lyons, Barbara Griffith, Terry Wu, Bob Sherwood, Julie Wilder, Julie Hutt, Michelle Valderas, Trevor Brasel, Dana Pohlman, Mitch Magee, Kathryn Sykes, Stephen Johnston, Karl Klose, Bernard Arulanandam, Justin Skoble ACRONYMS FOR TECHNICAL REPORTS ABSL3: Animal Biosafety Level 3 lab AM: alveolar macrophages ASU: Arizona State University BLD: below limit of detection BMDM: bone marrow derived macrophages BP: base pair BSL2: Biosafety Level 2 lab BSL3: Biosafety Level 3 lab CD14: cell marker for monocytes in humans and non-human primates CD16: cell marker for Natural killer cells in humans and non-human primates CD 56: cell marker for Natural killer cells in humans but may only be expressed in monocytes in non-human primates C: centrigrade cDNA: complimentary deoxyribonucleic acid CDA: confidentiality and disclosure agreement CDM: Chamberlains defined media CFSE: fluorescent dye called carboxyfluorescein succinimidyl ester CFU: colony forming units COA: contract officers authorization CRDA: Cooperative research and development agreement C-term: Carboxy terminus DNA: deoxyribonucleic acid DO: dissolved oxygen DVC: Dynport Vaccine Company ED50: Effective Dose, 50 % dead ELISA: enzyme-linked immunosorbent assay ELISpot: enzyme-linked immunospot assay EOH: Employee Occupational Health FACS: Fluorescence-activated cell sorting FDA: Federal Drug Administration FF: formalin fixed FSC: forward scatter, equivalent to cell size FT or Ft or F. tularensis: Francisella tularensis GDP: genome directed primers GI: gastrointestinal GLP: good laboratory practice KBMA: killed but metabolically active Page 3 of 6 Tularemia Vaccine Development Contract: Monthly Technical Report Period: 2/1/2010 to 2/28/2010 Due Date: 3/15/2010 and Prepared by: C. Rick Lyons, Barbara Griffith, Terry Wu, Bob Sherwood, Julie Wilder, Julie Hutt, Michelle Valderas, Trevor Brasel, Dana Pohlman, Mitch Magee, Kathryn Sykes, Stephen Johnston, Karl Klose, Bernard Arulanandam, Justin Skoble HK: heat killed HTP: high through put IAA: interagency agreement IACUC: Institutional animal care and use committee ICS: Intracellular staining ID: intradermal IFNγ: interferon gamma IgG: immunoglobulin, G class IgA: immunoglobulin, A class IgM: immunoglobulin, M class IL2: interleukin 2 IM: intramuscular IN: intranasal IND: investigational new drug IP: intraperitoneal IRB: institutional review board IRS: immune rat sera IT: intratracheal IV: intravenous IVT: in vitro translation/transcription KB: kilobase L: liter LAPT: Linear Amplification of Procaryotic Transpcript LBERI: Lovelace Biomedical and Environmental Research Institute LD50: Lethal dose 50% dead LD99: lethal dose, 99% dead Lm: Listeria monocytogenes LN: lymph node LPS: lipopolysaccharide LRRI: Lovelace Respiratory Research Institute LVS: Live vaccine strain (tularemia) NHP: nonhuman primate MAb: monoclonal antibody MDM: monocyte derived macrophages mL: milliliter MLN: mesenteric lymph node mM: Milli molar Page 4 of 6 Tularemia Vaccine Development Contract: Monthly Technical Report Period: 2/1/2010 to 2/28/2010 Due Date: 3/15/2010 and Prepared by: C. Rick Lyons, Barbara Griffith, Terry Wu, Bob Sherwood, Julie Wilder, Julie Hutt, Michelle Valderas, Trevor Brasel, Dana Pohlman, Mitch Magee, Kathryn Sykes, Stephen Johnston, Karl Klose, Bernard Arulanandam, Justin Skoble MOI: multiplicity of infection MS: Milestone MSCR: Milestone Completion Report MTA: material transfer agreement MTS reagent: (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2Htetrazolium, inner salt N: number NIAID: National Institute of Allergy and Infectious Disease NRS: normal rat sera N-term: amino- terminus OD: optical density OMP:outer membrane proteins ORF: open reading frame P60: constitutively expressed protein PBMC: peripheral blood mononuclear cells PBS: phosphate buffered saline PFA: paraformaldehyde pM: pico Molar pmol: picomole PCR: polymerase chain reaction QA: quality assurance QC: quality control qPCR: quantitative polymerase chain reaction RNA: ribonucleic acid RNU: T-cell deficient nude rats SC: subcutaneous SDS: sodium dodecyl sulfate SCHU S4: virulent strain of Francisella tularensis SIP: Special Immunizations Program at USAMRIID SOP: standard operating procedure SSC: side scatter, equivalent to cell density or granularity TBLN: tracheobronchial lymph node TFF:tangential flow filtration Tn: transposon TNFα: tumor necrosis factor alpha TVDC: Tularemia vaccine development contract uL: microliter UNM: University of New Mexico Page 5 of 6 Tularemia Vaccine Development Contract: Monthly Technical Report Period: 2/1/2010 to 2/28/2010 Due Date: 3/15/2010 and Prepared by: C. Rick Lyons, Barbara Griffith, Terry Wu, Bob Sherwood, Julie Wilder, Julie Hutt, Michelle Valderas, Trevor Brasel, Dana Pohlman, Mitch Magee, Kathryn Sykes, Stephen Johnston, Karl Klose, Bernard Arulanandam, Justin Skoble USAMMDA: United States Army Medical Materiel Development Activity USAMRIID: United States Army Medical Research Institute for Infectious Disease UTSA: University of Texas at San Antonio Vvm: volume of air per culture volume per minute WFI: water for injection WT: wild type Page 6 of 6