Cholera is an acute, water-borne diarrheal disease caused by the facultative Gram-negative bacterium, Vibrio cholerae of serogroup O1 of the classical and El Tor biotypes and by V. cholerae serogroup O139. The magnitude, severity and duration of recent outbreaks in Angola, Zimbabwe, Vietnam, and Haiti, have strengthened a recommendation by the World Health Organization for the preventive use of oral cholera vaccines to avert outbreaks and control their spread. Several live genetically attenuated cholera vaccine strains such as Peru-15 and V. cholerae 638 have been demonstrated to be clinically safe, immunogenic and protective in phase I and II clinical trials. However, the efficacy of current vaccines in an outbreak scenario has not been tested and could be challenged by the fact that their infectivity and potential for transmission to households and within the community are inferior to that of epidemic strains. Recent studies have shown that V. cholerae cells present in fresh cholera stools or within biofilm aggregates display a hyperinfective phenotype. The discovery of a V. cholerae hyperinfective physiological stage has led to a new epidemiological model for cholera that incorporates human-to-human transmission. We propose to develop a live, genetically-attenuated vaccine prototype based on strain 638 that could be administered as a hyperinfective biofilm to mimic cholera transmission in outbreaks, enhance immunogenicity, and promote herd immunity. We will first modify strain 638 by deleting the cholera phage attachment site to increase its safety without affecting its colonization capacity.
In Aim 1, we wil introduce additional mutations in this strain to enhance its infectivity, intestinal colonization, nd biofilm formation. Then, we will conduct competition assays in vivo to determine the capacity of planktonic and biofilm-derived cells of the modified vaccine to outcompete strain 638 for colonization of the suckling mouse small intestine.
In Aim 2, we will use an oral (intragastric) adult rabbit immunization model to compare the immunogenicity and protective capacity of planktonic and biofilm-derived hyperinfective vaccine candidates to that of the classical vaccine strain 638.

Public Health Relevance

The magnitude, severity and duration of recent cholera outbreaks have led the World Health Organization to recommend the preventive use of oral cholera vaccines to avert and control outbreaks. In this research effort, we will construct and evaluate improved vaccine candidates that could fulfill this goal.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21AI103693-01
Application #
8428416
Study Section
Vaccines Against Microbial Diseases (VMD)
Program Officer
Hall, Robert H
Project Start
2013-08-12
Project End
2015-07-31
Budget Start
2013-08-12
Budget End
2014-07-31
Support Year
1
Fiscal Year
2013
Total Cost
$270,018
Indirect Cost
$152,518
Name
Southern Research Institute
Department
Type
DUNS #
006900526
City
Birmingham
State
AL
Country
United States
Zip Code
35205
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Ayala, Julio C; Wang, Hongxia; Benitez, Jorge A et al. (2018) Molecular basis for the differential expression of the global regulator VieA in Vibrio cholerae biotypes directed by H-NS, LeuO and quorum sensing. Mol Microbiol 107:330-343
Wang, Hongxia; Silva, Anisia J; Benitez, Jorge A (2017) 3-Amino 1,8-naphthalimide, a structural analog of the anti-cholera drug virstatin inhibits chemically-biased swimming and swarming motility in vibrios. Microbes Infect 19:370-375
Ayala, Julio C; Silva, Anisia J; Benitez, Jorge A (2017) H-NS: an overarching regulator of the Vibrio cholerae life cycle. Res Microbiol 168:16-25
Silva, Anisia J; Benitez, Jorge A (2016) Vibrio cholerae Biofilms and Cholera Pathogenesis. PLoS Negl Trop Dis 10:e0004330
Benitez, Jorge A; Silva, Anisia J (2016) Vibrio cholerae hemagglutinin(HA)/protease: An extracellular metalloprotease with multiple pathogenic activities. Toxicon 115:55-62
Ayala, Julio C; Wang, Hongxia; Silva, Anisia J et al. (2015) Repression by H-NS of genes required for the biosynthesis of the Vibrio cholerae biofilm matrix is modulated by the second messenger cyclic diguanylic acid. Mol Microbiol 97:630-45
Ayala, Julio C; Wang, Hongxia; Benitez, Jorge A et al. (2015) RNA-Seq analysis and whole genome DNA-binding profile of the Vibrio cholerae histone-like nucleoid structuring protein (H-NS). Genom Data 5:147-150
Wang, Hongxia; Ayala, Julio C; Benitez, Jorge A et al. (2015) RNA-seq analysis identifies new genes regulated by the histone-like nucleoid structuring protein (H-NS) affecting Vibrio cholerae virulence, stress response and chemotaxis. PLoS One 10:e0118295