Most pathogenic bacteria express surface carbohydrates called capsular polysaccharides (CPSs). CPSs are important vaccine candidates given that they are located on the outermost surface of bacteria and they have distinct structures. These two features make them easily accessible and distinctly recognizable by immune surveillance, therefore resulting in production of CPS specific antibodies by B cells. To induce a CPS specific adaptive immune response (i.e., T cell-mediated B cell response), CPSs are conjugated with carrier proteins, and the conjugation products are called glycoconjugate vaccines. Due to insufficient understanding of their immune activation mechanisms, current glycoconjugate vaccine strategies have reached saturation and are largely modifications of past empirical conjugation methods. The production of the current generation of glycoconjugate vaccines is based on trial and error and does not make use of specific scientific knowledge to maximize stimulation of critical immune cells (i.e., helper T cells) involved in producing protective IgG antibodies. A new perspective to carbohydrate-based vaccine research is much needed. With the potential of establishing a new paradigm, our previous discovery and preliminary data demonstrate that the mammalian CD4+ T cell repertoire contains a population of carbohydrate-specific T cells (i.e., Tcarbs) that recognize carbohydrate epitopes. Here, we propose to expand on our previous discovery and define the molecular mechanisms for Tcarb activation by carbohydrate epitopes from a model glycoconjugate vaccine.
In Aim 1 of this proposal we will elucidate structural requirements for carbohydrate presentation by major histocompatibility complex class II (MHCII) proteins on the surface of antigen presenting cells (APCs) or as purified MHCII proteins via interaction studies.
In Aim 2 we will structurally and functionally characterize T cell receptor (TCR) recognition of glycan epitopes by Tcarbs. We believe our proposed studies will create a new platform to develop knowledge-based glycoconjugate vaccines that are enriched for functional Tcarb epitopes. Using the discovery of Tcarb activation mechanisms and of structures of glycan epitopes, we can design and develop new-generation glycoconjugate vaccines that will elicit strong and long lasting immune response to protect from bacterial infections.

Public Health Relevance

Infectious diseases caused by pathogenic bacteria have been a major threat to human health. Capsular polysaccharides (CPSs), by which most pathogenic bacterial surfaces are decorated, have been used as main components of glycoconjugate vaccines against bacterial diseases in clinical practice worldwide. Due to inadequate understanding of how the immune system interacts with these complex vaccines, their design and production have often been empirical, yielding poorly characterized, heterogeneous and variably immunogenic vaccines, with poor immune responses in high-risk populations such as the elderly and patients with weak immune systems. This project investigates how these vaccines stimulate responses in critical cells of the immune system. The structural and mechanistic knowledge attained in this project will have fundamental biomedical implications in producing future knowledge-based vaccines that are target-specific, structurally designed, highly immunogenic and protective, produced at much lower cost and allowing much wider use on a global scale than current vaccines to control or eliminate wide variety of infectious diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI123383-02
Application #
9285694
Study Section
Vaccines Against Microbial Diseases Study Section (VMD)
Program Officer
Gondre-Lewis, Timothy A
Project Start
2016-06-06
Project End
2021-05-31
Budget Start
2017-06-01
Budget End
2018-05-31
Support Year
2
Fiscal Year
2017
Total Cost
$375,000
Indirect Cost
$125,000
Name
University of Georgia
Department
Other Health Professions
Type
Organized Research Units
DUNS #
004315578
City
Athens
State
GA
Country
United States
Zip Code
30602
Sun, Lina; Ishihara, Mayumi; Middleton, Dustin R et al. (2018) Metabolic labeling of HIV-1 envelope glycoprotein gp120 to elucidate the effect of gp120 glycosylation on antigen uptake. J Biol Chem 293:15178-15194
Middleton, Dustin R; Zhang, Xing; Wantuch, Paeton L et al. (2018) Identification and characterization of the Streptococcus pneumoniae type 3 capsule-specific glycoside hydrolase of Paenibacillus species 32352. Glycobiology 28:90-99
Middleton, Dustin R; Lorenz, Walter; Avci, Fikri Y (2017) Complete Genome Sequence of the Bacterium Bacillus circulans Jordan Strain 32352. Genome Announc 5:
Sheikh, M Osman; Halmo, Stephanie M; Patel, Sneha et al. (2017) Rapid screening of sugar-nucleotide donor specificities of putative glycosyltransferases. Glycobiology 27:206-212
Middleton, Dustin R; Sun, Lina; Paschall, Amy V et al. (2017) T Cell-Mediated Humoral Immune Responses to Type 3 Capsular Polysaccharide of Streptococcus pneumoniae. J Immunol 199:598-603
Sun, Lina; Middleton, Dustin R; Wantuch, Paeton L et al. (2016) Carbohydrates as T-cell antigens with implications in health and disease. Glycobiology 26:1029-1040
Avci, Fikri Y (2016) A chicken vaccine to protect humans from diarrheal disease. Glycobiology 26:1137-1139
Li, Guoyun; Li, Lingyun; Xue, Changhu et al. (2015) Profiling pneumococcal type 3-derived oligosaccharides by high resolution liquid chromatography-tandem mass spectrometry. J Chromatogr A 1397:43-51