This project is directed to develop technology to identify antigens recognized by pathogen-specific class-l MHC restricted T cells by systematic screening of all pathogen protein coding genes, its ORFeome. The system is based on the observation that phagolysosomal destruction within antigen presenting cells (APCs) of E. co// expressing cytoplasmic listeriolysin (cytoLLO) results in leakage of phagosomal contents into the ARC cytosol and access of antigens to the host cell cytosolic MHC class I antigen processing pathway resulting in presentation of the antigen in the context of MHC class I molecules. We propose to focus on immunity to S. Typhi, a pathogen of great public health importance. Live, attenuated vaccine strains of Salmonella have the potential to be a cost-effective prophylactic measure in combating this disease, yet the antigens important in protection are largely unknown. Thus, the identification of putative S. Typhi proteins that might be involved in protection, by screening of the S. Typhi ORFeome, will allow us to explore our hypothesis that the increased efficacy of 4 doses of the oral Ty21 a typhoid vaccine as compared to a single dose is in part due to the repertoire of S. Typhi specific CD8+ T cells in vaccinees. One of the novel features of the proposed technology is an ability to confirm that S. Typhi proteins are expressed and able to be presented by APCs in the context of class-l molecules by fusing them with positive epitope controls. In addition, rather than co-expressing cytoLLO and a candidate antigen in the same ? co// cell, we propose to co-infect ARC with two different strains of E. co//, one expressing cytoLLO and the other the antigen of interest as this system allows greater flexibility in use of the ORFeome for other purposes. This proposal brings together collaborators with expertise in cellular immunology, genomics and high throughput assays systems with the express purpose of tackling an important global health problem via improved vaccine development.

Public Health Relevance

The development of improved typhoid vaccines to prevent antibiotic-resistant typhoid fever in developing countries is a high global public health priority. In this project we propose to examine the whole ORFeome of S. Typhi to uncover proteins that are recognized by the human host using cells from immunized subjects. Findings from these studies will greatly contribute to accelerate the development of typhoid vaccines.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program--Cooperative Agreements (U19)
Project #
5U19AI082655-04
Application #
8378488
Study Section
Special Emphasis Panel (ZAI1-KS-I)
Project Start
Project End
Budget Start
2012-06-01
Budget End
2013-05-31
Support Year
4
Fiscal Year
2012
Total Cost
$310,558
Indirect Cost
$97,136
Name
University of Maryland Baltimore
Department
Type
DUNS #
188435911
City
Baltimore
State
MD
Country
United States
Zip Code
21201
Salerno-Goncalves, R; Safavie, F; Fasano, A et al. (2016) Free and complexed-secretory immunoglobulin A triggers distinct intestinal epithelial cell responses. Clin Exp Immunol 185:338-47
Fresnay, Stephanie; McArthur, Monica A; Magder, Laurence et al. (2016) Salmonella Typhi-specific multifunctional CD8+ T cells play a dominant role in protection from typhoid fever in humans. J Transl Med 14:62
Blohmke, Christoph J; Darton, Thomas C; Jones, Claire et al. (2016) Interferon-driven alterations of the host's amino acid metabolism in the pathogenesis of typhoid fever. J Exp Med 213:1061-77
Salerno-Goncalves, Rosangela; Fasano, Alessio; Sztein, Marcelo B (2016) Development of a Multicellular Three-dimensional Organotypic Model of the Human Intestinal Mucosa Grown Under Microgravity. J Vis Exp :
McArthur, Monica A; Fresnay, Stephanie; Magder, Laurence S et al. (2015) Activation of Salmonella Typhi-specific regulatory T cells in typhoid disease in a wild-type S. Typhi challenge model. PLoS Pathog 11:e1004914
Trebicka, Estela; Shanmugam, Nanda Kumar N; Chen, Kejie et al. (2015) Intestinal Inflammation Leads to a Long-lasting Increase in Resistance to Systemic Salmonellosis that Requires Macrophages But Not B or T Lymphocytes at the Time of Pathogen Challenge. Inflamm Bowel Dis 21:2758-65
Wahid, R; Fresnay, S; Levine, M M et al. (2015) Immunization with Ty21a live oral typhoid vaccine elicits crossreactive multifunctional CD8+ T-cell responses against Salmonella enterica serovar Typhi, S. Paratyphi A, and S. Paratyphi B in humans. Mucosal Immunol 8:1349-59
Booth, Jayaum S; Salerno-Goncalves, Rosangela; Blanchard, Thomas G et al. (2015) Mucosal-Associated Invariant T Cells in the Human Gastric Mucosa and Blood: Role in Helicobacter pylori Infection. Front Immunol 6:466
Toapanta, Franklin R; Bernal, Paula J; Fresnay, Stephanie et al. (2015) Oral Wild-Type Salmonella Typhi Challenge Induces Activation of Circulating Monocytes and Dendritic Cells in Individuals Who Develop Typhoid Disease. PLoS Negl Trop Dis 9:e0003837
Sztein, Marcelo B; Salerno-Goncalves, Rosangela; McArthur, Monica A (2014) Complex adaptive immunity to enteric fevers in humans: lessons learned and the path forward. Front Immunol 5:516

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