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Principal Investigator: Paola Massari
Organization: TUFTS UNIVERSITY BOSTON
Fiscal Year: 2019
Award: $247,500
Funding agency: National Institute of Allergy and Infectious Diseases
Project Summary
Neisseria gonorrhoeae reproductive tract infections can lead to chronic and severe consequences in women,
where diagnosis is frequently delayed by lack of symptoms, contrary to male infections. A once-simple
antibiotic treatment gonococcal infection is now compromised by a worldwide increased antibiotic resistance,
including the last FDA-approved antibiotic cefixime. The CDC has recently placed N. gonorrhoeae in the urgent
threat category of antibiotic resistant microorganisms. There is a strong need for a vaccine against gonorrhea,
but this has been delayed by combined lack of successful antigens and animal models of N. gonorrhoeae
infections to evaluate protective immunity. Discovery of new bacterial antigens has been enabled by the
assumption that pathogens express virulence factors explicitly in vivo. These may have consequently escaped
detection in vitro. Based on previous NIH-funded grants (U19 AI084048 and R56 AI107821-01A1), we have
examined the global transcriptome of cervico-vaginal lavage and urethral samples from human subjects
naturally infected with N. gonorrhoeae attending the National Center for STD Control (NCSTD) in Nanjing,
China, by RNA-seq. We reported a differential mRNA expression in these samples as compared to that from
each infecting strains grown in vitro, for a large number of genes. We have defined these “in vivo expressed
factors” (IVEFs). Among the IVEFs there are several hypothetical proteins, representing a previously untapped
pool of potential new antigens for a N. gonorrhoeae vaccine. These novel IVEFs vaccine candidates have not
been examined before because they had not been identified so far. We have used multiple in silico-based
predictive criteria to select several hypothetical protein IVEFs to be tested in vitro and in vivo as novel broad
vaccine targets. Besides being highly expressed in vivo, the desired features of the selected IVEFs include:
broad expression among gonococcal strains; predicted antigenicity; predicted bacterial membrane localization
(outer membrane or periplasmic) and surface exposure and lastly, structure features suitable for recombinant
production. In addition to predicted antigenicity, some IVEFs share DNA sequence homology to currently
explored immunogenic meningococcal vaccine candidates. We will initially focus on 19 IVEFs for recombinant
expression and antigenic/immunogenic characterization in mice. Immunogenicity as well as recognition of
IVEFs in whole bacteria will be validated using banked hyperimmune sera from subjects naturally exposed to
the gonococcus, for which we will use purified IVEFs as well as IVEF over-expression gonococcal mutant
strains created in both laboratory strains and clinical isolates from the Nanjing cohort. We will test the ability of
mouse immune sera and vaginal lavages as well as banked human sera to induce killing of N. gonorrhoeae in
vitro to further establish IVEFs as broad vaccine antigens.
Terms: <7S Gamma Globulin><ATGN><Adjuvant><Alum Adjuvant><Animal Model><Animal Models and Related Studies><Animals><Antibiotic Agents><Antibiotic Drugs><Antibiotic Resistance><Antibiotic Therapy><Antibiotic Treatment><Antibiotics><Antibodies><Antibody Specificity><Antigens><Antisera><Apoptosis Response Protein><BALB C Mouse><BALB/c><Bacteria><Bacterial Antigens><Bio-Informatics><Biochemical><Bioinformatics><CDC><Cannot achieve a pregnancy><Categories><Cefixime><Centers for Disease Control><Centers for Disease Control and Prevention><Centers for Disease Control and Prevention (U.S.)><China><Chronic><Clinical><Collaborations><Computer Models><Computer Simulation><Computer based Simulation><Computerized Models><DNA Sequence><Detection><Diagnosis><Difficulty conceiving><Disease><Disorder><Douching><Drug Resistance in N. Gonorrhoeae><Drug resistance in Neisseria Gonorrhoeae><Drug resistant N. Gonorrhoeae><Drug-resistant N. Gonorrhoeae><Drug-resistant Neisseria Gonorrhoeae><E coli><E. coli><Ectopic Pregnancy><Escherichia coli><Evaluation><Exposure to><FDA approved><Female><Funding><Gene Expression><Gene Transcription><Genes><Genetic Transcription><Genital><Genital system><Goals><Gonococcal Infection><Gonococcus><Gonorrhea><Grant><Health Sciences><Human><Hypothetical Protein><IgA><IgG><Immune Sera><Immune response><Immunity><Immunize><Immunochemical Immunologic><Immunoglobulin A><Immunoglobulin G><Immunologic><Immunological><Immunological response><Immunologically><Immunologics><In Vitro><Inbred BALB C Mice><Infection><Infertility><Irrigation><Laboratories><Lavage><Lead><Letters><Mainland China><Mathematical Model Simulation><Mathematical Models and Simulations><Measures><Membrane><Meningococcal vaccine><Messenger RNA><Mice><Mice Mammals><Miscellaneous Antibiotic><Modern Man><Morbidity><Morbidity - disease rate><Mucosa><Mucosal Tissue><Mucous Membrane><Murine><Mus><N gonorrhea><N gonorrhoeae><N. gonorrhea><N. gonorrhoeae><N.gonorrhoeae><NIH><National Institutes of Health><Neisseria gonorrhea><Neisseria gonorrhoeae><Nonvaginal irrigation><Nonvaginal lavage><Organism><PAWR protein><PRKC, Apoptosis, WT1, Regulator><Pathogenicity><Pathogenicity Factors><Pb element><Pelvic Inflammatory Disease><Periplasmic Space><Pharmacology><Production><Prostate Apoptosis Response Protein 4><Proteins><RNA Expression><RNA Seq><RNA sequencing><RNAseq><Recombinants><Reporting><Reproductive Tract Infections><Research Specimen><Resistance to antibiotics><Resistant to antibiotics><Sampling><Sequence Homology><Sexually Transmitted Agents><Sexually Transmitted Diseases><Sexually Transmitted Disorder><Sexually Transmitted Infection><Specimen><Structure><Surface><Symptoms><Tbp1 Transferrin-Binding Protein><TbpA Transferrin-Binding Protein><Testing><Therapeutic><Transcript><Transcription><Transcriptional Repressor PAR4><Transferrin-Binding Protein A><United States Centers for Disease Control><United States Centers for Disease Control and Prevention><United States National Institutes of Health><University Health Services><Urethra><Vaccine Antigen><Vaccines><Vaginal Douching><Vaginal Irrigation><Vaginal Lavage><Venereal Diseases><Venereal Disorders><Venereal Infections><Virulence Factors><WT1-Interacting Protein><Whole Organism><Woman><Work><alum><aluminum sulfate><antibiotic drug resistance><antibiotic resistant><antibiotic resistant Neisseria gonorrhoeae><bacterial disease treatment><bacterial infectious disease treatment><bactericidal><bactericide><base><cervicovaginal><cohort><comparative><computational modeling><computational models><computational simulation><computer based models><computerized modeling><computerized simulation><develop a vaccine><development of a vaccine><experiment><experimental research><experimental study><extrauterine pregnancy><fertility cessation><fertility loss><genital tract><global gene expression><global transcription profile><gonorrhea vaccine><heavy metal Pb><heavy metal lead><host response><human subject><immune serum><immunogen><immunogenic><immunogenicity><immunoresponse><in silico><in vitro Assay><in vitro testing><in vivo><in vivo evaluation><in vivo testing><infertile><irrigation therapy><lavage therapy><living system><mRNA><mRNA Expression><male><membrane structure><microorganism><model of animal><model organism><mouse model><murine model><mutant><new vaccines><next generation vaccines><novel><novel vaccines><other than vaginal Douching><other than vaginal Irrigation><overexpress><overexpression><par-4 protein><pathogen><pelvic inflammatory syndrome><periplasm><periplasmic><prevent><preventing><protein expression><reproductive tract><transcriptome><transcriptome sequencing><urethral><vaccine candidate><vaccine development><vaccine evaluation><vaccine formulation><vaccine screening><vaccine testing><virtual simulation>