Source: PURDUE UNIVERSITY submitted to
OPTIMIZING THE IMMUNE RESPONSE TO ALUMINUM-CONTAINING VACCINES
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
0229459
Grant No.
(N/A)
Project No.
IND020164H
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Oct 1, 2012
Project End Date
Sep 30, 2017
Grant Year
(N/A)
Project Director
Hogenesch, HA.
Recipient Organization
PURDUE UNIVERSITY
(N/A)
WEST LAFAYETTE,IN 47907
Performing Department
Veterinary Comparative Pathobiology
Non Technical Summary
Aluminum adjuvants are widely used in veterinary and human vaccines. The proposed research is aimed at two approaches to increase the immune response to aluminum adjuvanted vaccines. The first is to modify the interaction between aluminum adjuvants and antigens by adding phosphate groups to the vaccine antigens. This would lead to prolonged retention of antigen at the injection site. The second approach is to change the type of inflammation at the injection site. The inflammation is an important determinant of the immune response that develops. This research may result in new vaccines and in improvements of existing vaccines by dose sparing of vaccine antigens and a reduction in the number of doses needed to induce a protective immune response.
Animal Health Component
25%
Research Effort Categories
Basic
(N/A)
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3083999109025%
3113999109025%
7123999109025%
7223999109025%
Knowledge Area
311 - Animal Diseases; 712 - Protect Food from Contamination by Pathogenic Microorganisms, Parasites, and Naturally Occurring Toxins; 722 - Zoonotic Diseases and Parasites Affecting Humans; 308 - Improved Animal Products (Before Harvest);

Subject Of Investigation
3999 - Animal research, general;

Field Of Science
1090 - Immunology;
Goals / Objectives
The overall goal of this research is to develop more effective formulations of aluminum adjuvanted vaccines. The specific objectives of this research are: 1.Optimize the interaction between antigens and aluminum adjuvant particles by modifying the antigens with a linker agent terminal phosphate groups. Terminal phosphate groups can exchange with hydroxyl groups at the surface of the aluminum adjuvants resulting in a stable bond. By varying the number of linkers and the length of the linkers, the interaction between antigen and adjuvant can be changed and optimized. 2.Determine the kinetics of cell infiltration and gene expression at the injection site following intramuscular injection of the aluminum-adjuvanted vaccines. 3.Determine the effect of manipulation of the inflammatory response at the injection site on the subsequent immune response. Selective depletion of types of inflammatory cells and blocking of specific cytokines may result in a qualitatively and quantitatively enhanced immune response. Information obtained through these studies will be published in peer-reviewed journals, presented at national and international meetings and may generate commercial products.
Project Methods
Objective 1. Linkers will be prepared that vary in length and composition, and the number of linkers per antigen molecule will be varied. This work will initially be carried out with two model antigens, hen egg lysozyme (HEL) and a genetically modified inactive form of diphtheria toxin, CRM197. Other vaccine antigens may be included as they become available. The interaction between adjuvants and modified antigens will be determined in vitro by measuring the adsorption. The effect of the linker modification on the antibody response will be determined in a mouse model. Geometric mean titers will be calculated for each group. A one-way ANOVA will be performed on log2-transformed titers to determine if there are significant differences between groups. If a significant difference is detected at P < 0.05, a Bonferroni multiple comparison post-hoc analysis will be performed to compare the differences between groups. Objective 2.A model vaccine composed of aluminum hydroxide adjuvant with ovalbumin will be prepared. The vaccine will be injected into the gastrocnemius muscle of mice. Injection sites will be collected 6 hours, 1 day, 2 days, and 7 days after injection. In some experiments, the mice will first be immunized intraperitoneally with ovalbumin or sterile saline (as a control) followed 3 weeks later by intramuscular injection of the aluminum-adjuvanted vaccines. The kinetics and type of inflammation at the injection site in naive and previously immunized mice will be compared by immunofluorescence and RT-PCR. The significance of differences in histology scores between groups will be determined by nonparametric analysis of variance (Kruskall Wallis) followed by Dunn's multiple comparison test. Differences will be considered significant at P < 0.05. For mRNA expression the geometric means of fold increases in mRNA expression will be compared on log-2 transformed data by one-way ANOVA followed by a Bonferroni multiple comparison post-hoc analysis. Objective 3. Monoclonal antibody-based depletion and genetically engineered mice will be used to determine the role of specific cell types. The target cell type will depend on the results of the experiments in Objective 2.

Progress 10/01/12 to 09/30/17

Outputs
Target Audience:Scientists in academia and industry working on vaccine discovery and development. Students. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project supported a graduate student who graduated with a PhD in December 2016. In addition, two undergraduate students, one majoring in Biological Science/pre-vet and one in Animal Sciences/pre-vet are working on this project for course credits. How have the results been disseminated to communities of interest?Via journal publications and conference presentations What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Adjuvants are substances added to vaccines to boost the immune response. Aluminum-containing adjuvants are commonly used in human and veterinary vaccines. They have an excellent safety record and are effective in stimulating the immune response against many, but not all infectious diseases. Our research is aimed at improving the performance of aluminum adjuvants while maintaining the safety profile, and on developing new adjuvants that have certain benefits not found in aluminum adjuvants. We developed new methods to control the adsorption of vaccine antigens to aluminum adjuvants. Adsorption is important as it increases retention of antigens at the injection site and allows more time for recruited antigen-presenting cells to take up antigens and initiate the immune response. We designed and synthesized linker molecules that increase adsorption taking advantage of the high affinity of aluminum for phosphate. We also investigated whether the efficay of aluminum adjuvants could be enhanced by combining aluminum adjuvants with other immunostimulatory molecules. Indeed, adsorption of certain Toll-like receptor agonists stimulated a robust immune response. Aluminum adjuvants have certain drawbacks, including the susceptibility to freezing, destabilization of certain vaccine antigens, and incompatibility with mucosal routes of vaccine delivery. We developed a novel nanoparticle adjuvant based on phytoglycogen derived from a variety of sweet corn. Positively charged nanoparticles significantly stimulated the immune response. The nanoparticles are stable, relatively inexpensive, and can be used for intranasal vaccine delivery as well as injectable vaccines. They appear to have great potential as an alternative adjuvant for food animal vaccines.

Publications

  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Lu F, Mosley Y, Rosales RR, Carmichael BE, Elesela S, Yao Y, HogenEsch H (2017) Alpha-D-glucan nanoparticulate adjuvant induces a transient inflammatory response at the injection site and targets antigen to migratory dendritic cells. Npj Vaccines 2: 4, 2017
  • Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: HogenEsch H. Chemistry and Immunobiology of Aluminum-containing adjuvants. CRWAD meeting, Chicago, 2016. Invited presentation.
  • Type: Theses/Dissertations Status: Awaiting Publication Year Published: 2017 Citation: Lu F. Evaluation of dendrimer-like alpha-D-glucan nanoparticles as a vaccine adjuvant.


Progress 10/01/15 to 09/30/16

Outputs
Target Audience:Scientists in academia and industry working on vaccine discovery and development. Students. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project supports a graduate student. In addition, an undergraduate student majoring in Biological Science and pre-vet, is working on this project for course credits. How have the results been disseminated to communities of interest?Presentation at an international meeting and peer-reviewed journal article. What do you plan to do during the next reporting period to accomplish the goals?We plan to further investigate the mechanism by which TLR agonists enhance the immune response when administered in combination with aluminum adjuvants. We will also investigate the effect of Nano-11 nanoparticles on uptake of antigens by antigen-presenting cells and conduct further studies to investigate their potential as an alternative to aluminum and oil-based adjuvants.

Impacts
What was accomplished under these goals? The effect of combining aluminum adjuvants with TLR agonists on the transport of antigens to the draining lymph nodes was investigated in a mouse model. We also continued the studies of alpha-D-glucan dendrimer-like nanoparticles. The distribution of the nanoparticles in vivo and the effect on the transport of antigen to the draining lymph nodes was determined by in vivo imaging. We also determined the effect of the nanoparticles on the immune response to intranasally administered antigens.

Publications

  • Type: Journal Articles Status: Accepted Year Published: 2016 Citation: HogenEsch H, Dunham A, Burlet E, Lu F, Mosley YYC, Morefield G (2016) Preclinical safety study of a recombinant Streptococcus pyogenes vaccine formulated with aluminum adjuvant. J Applied Toxicol, in press
  • Type: Conference Papers and Presentations Status: Other Year Published: 2016 Citation: Corn-derived ?-D-glucan nanoparticles are safe and effective vaccine adjuvants. Biomarkers for Vaccine and Adjuvant Potency and Safety. Porto, Portugal, 2016.


Progress 10/01/14 to 09/30/15

Outputs
Target Audience:Scientists in academia and industry working on vaccine discovery and development. Students. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project supports a graduate student. In addition, an undergraduate student majoring in Biological Science and pre-vet, is working on this project for course credits. How have the results been disseminated to communities of interest?Via a journal publication. What do you plan to do during the next reporting period to accomplish the goals?We plan to further investigate the effect of TLR agonists in combination with aluminum adjuvants, and to investigate the potential of the Nano-11 nanoparticles as an alternative to aluminum and oil-based adjuvants. The efficacy of Nano-11 for mucosal vaccination will also be investigated.

Impacts
What was accomplished under these goals? We continued to work on investigating the optuimal formulation and benefits of combining aluminum adjuvants with TLR-agonists.In addition, we investigated the potential immunostimulating effect of naturally occurring nanoparticles in a variety of sweet corn. These alpha-D-glucan nanoparticles can be modified to give them a positive surface charge. The positively charged nanoparticles, termed Nano-11, enhanced the immune response while inducing a limited and transient inflammatory reaction at the site of injection. The nanoparticles are inexpensive and a potential alternative to currently used adjuvants in animal health vaccines.

Publications

  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Lu F, Mencia A, Bi L, Taylor A, Yao Y, HogenEsch H (2015) Dendrimer-like alpha-D-glucan nanoparticles activate dendritic cells and are effective vaccine adjuvants. J Control Release, 204: 51-59


Progress 10/01/13 to 09/30/14

Outputs
Target Audience: Scientists who read published articles Students Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? This project supports the training of a graduate student. In addition, amedical student worked on this project during the summer of 2014. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? We plan to determine the effect of TLR agonists on the activation of dendritic cells in vitro, and on the recruitment and migration of dendritic cells in vivo.

Impacts
What was accomplished under these goals? The effect of combining aluminum adjuvants with Toll-like receptor agonists on the inflammatory response and immune response was determined. The addition of TLR agonists enhanced the production of IgG2a antibodies (Th1 response), while maintaining the production of IgG1 antibodies (Th2 response). The number of eosinophils in the injectionsite actually increased upon addition of TLR agonists while the number of MHCII+ cells decreased.

Publications

  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Morefield G, Touhey G, Lu F, Dunham A, HogenEsch H (2014) Development of a vaccine to protect against Streptococcus pyogenes infection. Vaccine, 32:3810-3815


Progress 10/01/12 to 09/30/13

Outputs
Target Audience: Readers of published articles. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? A graduate student is working on this research project. How have the results been disseminated to communities of interest? Through peer-reviewed publications. What do you plan to do during the next reporting period to accomplish the goals? 1.In collaboration with Dr. Borch, develop linkers tobind carbohydrate antigens to aluminum adjuvants 2. Determine if manipulation of the inflammatory response at the injection site affects the immune response to aluminum adjuvanted vaccines.

Impacts
What was accomplished under these goals? 1. A phosphonate linker was developed that allowed controlledbinding of a model protein, lysozyme, to aluminum adjuvant. Using this linker, we demonstrated that adsorption is critical at low antigen doses and not necessary at higher doses. 2. The kinetics of inflammatory cell accumulation at the injection site is influenced by prior immunization. 3. Although neutrophils are the first inflammatory cells to arrive at the injection site in large numbers, depletion of neutrophils did not affect the immune response to an aluminum-adjuvanted vaccine.

Publications

  • Type: Journal Articles Status: Published Year Published: 2013 Citation: HogenEsch H (2013) Mechanism of immunopotentiation and safety of aluminum adjuvants. Front Immunol, 3:406.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Lu F, HogenEsch H (2013) Kinetics of inflammation following injection of aluminum-adjuvanted vaccines. Vaccine, 31: 3979-3986.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Lu F, Boutselis I, Borch RF, HogenEsch H (2013) Control of antigen-binding to aluminum adjuvants and the immune response with a novel phosphonate linker. Vaccine, 31:4362-4367.
  • Type: Book Chapters Status: Published Year Published: 2013 Citation: Mittal SK, HogenEsch H, Vemulapalli R, Park K (2013) Vaccines, adjuvants and delivery systems for infectious diseases. In: Encyclopedia of Pharmaceutical Technology. 4th edition. Swarbrick J, ed. Informa Healthcare.