Source: UNIVERSITY OF WYOMING submitted to
NK CELL MEMORY IN LONG-TERM IMMUNITY TO TOXOPLASMA GONDII.
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
Reporting Frequency
Annual
Accession No.
1007951
Grant No.
(N/A)
Project No.
WYO-563-16
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Oct 22, 2015
Project End Date
Sep 30, 2017
Grant Year
(N/A)
Project Director
Gigley, JA.
Recipient Organization
UNIVERSITY OF WYOMING
1000 E UNIVERSITY AVE DEPARTMENT 3434
LARAMIE,WY 82071-2000
Performing Department
Molecular Biology
Non Technical Summary
The parasite Toxoplasma gondii (T. gondii) is the third leading cause of food borne illness in the U.S. Infection rates range between 30-70% in many animal species, including livestock, and as such is a major food safety issue world wide. In humans, there are over 1,075,242 primary infections per year caused mostly by eating contaminated meat products with annual cost of illness of 3 billion dollars. In addition to the food safety threat, it is also one of the top reasons behind infectious abortion in livestock, especially sheep and goats resulting in significant losses in annual animal productivity. Infection of pigs, chickens, goats, sheep and cattle occurs when their food and water is contaminated by the excreted cat form of the parasite known as the oocyst. To date, no effective vaccines or drug treatments exist to completely protect against or clear T. gondii infection in livestock. In Europe, one attempt has been made to produce a safe and effective ovine vaccine against T. gondii. This treatment (Toxovax) was only partially effective and lasted for only 18 months. Therefore, development of a safe and effective vaccine for livestock is essential to improve animal health and livestock production while also improving food safety by reducing transmission to humans and thus, food-borne illness associated with this pathogen.Although a T cell population, known as the CD8 T cell, is widely recognized as a key player in long-term control of infection, a recent study using a new Toxoplasma vaccine strain identified a second important type of cell, the natural killer (NK) cell for immunity. NK cells are classically thought to be part of the early immune response, known as the innate immune response. They are effective against cancer and infectious disease and classically not thought to last a long time. This viewpoint has started to change. Recent studies suggest NK cells may become long-lived and be potent contributors to long-lasting immune responses, known as adaptive immune responses. This is important for vaccine development for livestock because if we can understand how to make them have memory to a pathogen, they would be useful in protecting against infection and increase the efficacy of the vaccine. A major obstacle in trying to stimulate NK cells to have memory is that the factors required to make them develop this trait are not well known. We now have found that NK cells are absolutely required for long-lasting immunity to Toxoplasma gondii infection. We do not understand how this occurs. The goal of this project is to define how they develop this response so we can better design vaccines to improve animal health, productivity and improve food safety.
Animal Health Component
100%
Research Effort Categories
Basic
100%
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
7123999109050%
3114099111025%
3134099109025%
Goals / Objectives
Currently, no vaccines or drugs are capable of adequately treating Toxoplasma in livestock. An effective vaccine for T. gondii in agricultural animals should accomplish two goals: 1) reduce infection induced abortions, and 2) Reduce or eliminate parasite tissue burdens in food animals to prevent food source contamination. Good vaccine design relies on a good knowledge base. To this end, our major goal is to increase knowledge of immune system function required for optimal immunity to Toxoplasma sowe can design a highly effective vaccine or treatment to improve animal health. Specifically, we will dissect the role of Natural Killer cells (NK) in adaptive immunity to T. gondii and define mechanism(s) behind this functional development. To improve agriculture animal health and prevent infection induced abortions caused by the parasite Toxoplasma gondii. We will investigate how the Natural Killer cell plays a role in long lasting immune responses to this pathogen. We will then dissect the mechanisms by which they develop this activity. Achieving the goals will improve vaccine and therapy efficacy to treat this infection.
Project Methods
The methods for this project will include a mouse based model of infection with Toxoplasma gondii. We will also heavily use flow cytometry to characterize and determine what the Natural Killer cells are doing during different times of infection. We will use antibody treatments and transgenic animals to follow the fate of natural killer cells over time. All methodologies are standard immunology and cell biology techniques. Specific methods are included in the attached proposal.

Progress 10/22/15 to 09/30/17

Outputs
Target Audience:Our target audience are individuals who study immunology and parasitology. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project has provided significant training and professional development for myself and one of my graduate students. The opportunities have been the ability to publish data in a peer review journal and attend meetings to present data on the subject of interest. How have the results been disseminated to communities of interest?Results have been disseminated to communities of interest via publication and meetings. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? We accomplished several parts of the proposed project during the funding period. The first part we achieved was to demonstrate formally that NK cells contribute significantly to adaptive immunity against T. gondii. This alone is a major new finding because these cells were only thought to contribute to early control of the parasite during primary infection and not during a secondary challenge. This is important because it highlights that from a vaccine standpoint the NK cells are a required component of a vaccine induced response. The second part we achieved has been to elucidate the mechanisms of how NK cells are able to mount a second response and how they contribute to parasite control in secondary exposure. NK cells produce IFN gamma which is a cytokine critical in controling Toxoplasma infection. The cytokine IL-12 has long been considered to be the only cytokine important in activating NK cells to produce IFN gamma. Our data now demonstrate that in a vaccine rechallenge model, Il-12 is required, but another IL-12 family member IL-23 is also required for NK cell IFN gamma. IL-23 has never been considered to induce IFN gamma production. This is important to show because IL-23 not seems to be an important piece of a vaccine induced response. Dissecting how NK cells contribute to control of the parasite we have determined that NK cells most likely act alone via their IFN gamma production. Taken together we now understand that NK cells are important in adaptive immunity and they require the cytokine IL-23 and IL-12 to carry out their function.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: 1. Comparative analysis of conventional natural killer cell responses to acute infection with Toxoplasma gondii strains of different virulence. J. Immunol. AAI meeting 2017
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: 2. NK cells in chronic Toxoplasma gondii infection: Friend or Foe. Immunology and Immunotherapy symposium, Geisel School of Medicine, Dartmouth College. March 2017
  • Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: 3. Natural Killer Cells: Friend or Foe in Chronic Toxoplasmosis. International Coccidiosis Conference. July 2017.


Progress 10/22/15 to 09/30/16

Outputs
Target Audience:Our target audience are investigators who are interested in innate immune responses to infection. In particular people who are interested in how Natural killer cells respond to parasitic infection and contribute to long term immunity against them. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project has provided extensive support and training for my graduate student to carry out her experiments. She will attend an international meeting and present some of her published work while there. How have the results been disseminated to communities of interest?We have published two papers currently and have another in the works. What do you plan to do during the next reporting period to accomplish the goals?During the next reporting cycle we will define the extrinsic mechanism that regulates the NK cell response to secondary infection. We will also determine the NK cell function required for protection against secondary parasite infection. We have putatively identified a receptor on NK cells that may be involved in receiving the activating signal to generate the secondary NK cell response. We will publish this work.

Impacts
What was accomplished under these goals? 1. We have cleary demonstrated that NK cells are required for vaccine induced protection against Toxoplasma gondii. 2. We have determined that NK cells are not intrinsically different after vaccination than prior to vaccination and thus do not appear to be further differentiated into long live memory cells. 3. We have shown that a cell extrinsic process is involved and are further pursuing what extrinsic factor is causing the need for NK cells in adaptive immunity.

Publications

  • Type: Journal Articles Status: Published Year Published: 2016 Citation: 1. Ivanova, D., Fatima, R. and Gigley, J.P. (2016) Comparative analysis of conventional natural killer cell responses to acute infection with Toxoplasma gondii strains of different virulence. Frontiers In Immunol. 2. Gigley, J.P. (2016) The Diverse Role of NK cells in immunity to Toxoplasma gondii infection. PLOS Pathogens. Feb. 25;12(2):e1005396