Source: MONTANA STATE UNIVERSITY submitted to
ANALYSIS OF GAMMA/DELTA T CELLS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
REVISED
Funding Source
Reporting Frequency
Annual
Accession No.
0158522
Grant No.
(N/A)
Project No.
MONB00418
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Oct 1, 2010
Project End Date
Sep 30, 2015
Grant Year
(N/A)
Project Director
Jutila, MA.
Recipient Organization
MONTANA STATE UNIVERSITY
(N/A)
BOZEMAN,MT 59717
Performing Department
Microbiology & Immunology
Non Technical Summary
Effective stimulation of the immune system by vaccines is dependent upon the proper targeting of appropriate immune cells (lymphocytes). Gamma/delta T cells represent the major T cell subset in newborn calves, yet we know little of their function and how and when to properly stimulate these cells for long lasting immunity. The purpose of this project is to study the bovine gamma/delta T cells using functional, biochemical, and molecular approaches. The direct impact of this work is on our general understanding of the bovine immune system and how it is similar and dissimilar to the immune system in rodents and humans. This information should facilitate development of new approaches to treat infectious diseases of cattle.
Animal Health Component
35%
Research Effort Categories
Basic
65%
Applied
35%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3153410109025%
3153410104025%
3113410109025%
3113410104025%
Goals / Objectives
This project provides partial salary support for personnel in the laboratory. Research operations come from funded, competitive grants. In this renewal, we plan on continuing our studies of bovine gamma/delta T cells by focusing on studies of unique gene expression in these cells, as well as, in the development of approaches to control their activity to the ultimate benefit of the animal. Specifically, we hypothesize that NRAMP-1 in γδ T cells enhances their response to PAMPS, cytokines, and/or other unique agonists. Specific Aim 1. Define NRAMP-1 expression in γδ T cells. Specific Aim 2. Examine the effects of cellular activation on expression of NRAMP-1, including potential alterations in intracellular localization and association with membrane receptors. Specific Aim 3. Test the role of NRAMP-1 in enhancing signaling responses in bovine γδ T cells. Specific Aim 4. Continued analysis of myeloid cell gene expression and specific agonist responses in bovine γδ T cells
Project Methods
Our primary source of cells will be from the blood of 1-6 month old calves. Using standard immunological and biochemical approaches, expression of NRAMP-1 will be confirmed under steady state and activated conditions. The main immunological assays for these experiments will be immunohistology and SDS/PAGE (Western blots). Cell culture techniques, combined with the use of a variety of materials that activate gamma/delta T cells, will be used to follow changes in NRAMP-1 expression. Functional studies will use RNAi approaches to "knock-down" NRAMP-1 transcripts and examine the functional consequences to the cell. Continued screens for novel materials that activate bovine gamma/delta T cells will be done, predominantly by use of multi-color flow cytometry. Gene expression will be studied by RT-PCR, microarrays, and modifications of SAGE. All results will be confirmed in replicate experiments using appropriate statistical approaches. The main target audience for this work will be investigators in the field and the main approaches for disseminating this information will be presentations at scientific meetings and publication of peer-reviewed papers. The progress in this project will be evaluated based on the number of peer-reviewed publications and competitive grants supporting the operation costs of the laboratory.

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

Outputs
Target Audience: Primary target are investigators in the field of animal health. Also, livestock producers are the end target of future translational efforts in this project. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? The project contributed to the PhD dissertation work on one student and MS work of another. How have the results been disseminated to communities of interest? Yes, through publications and presentations at scientific meetings. Per the latter, we presented multiple abstracts at the national AAI meeting and submitted abstracts to the Annual USDA/NIF Project Directors meeting and a Keystone meeting in January, 2015. What do you plan to do during the next reporting period to accomplish the goals? This core projects supports the basic operation of my laboratory, which supports three grant funded programs: NIH funded R21 focused on the study of innate responses during C. burnetii infection, USDA/NIFA funded project focused on testing the effects of amphotericin B in innate immunity in calves, and a USDA Animal Health grant on testing the effects of a plant derived polysaccharide on innate immunity in calves. Each of those projects will continue into the coming year. Another major effort in the coming year will be to prepare a renewal of this project for review by the College of Agriculture at MSU for potential funding in 2015. This renewal will, again, focus on all projects in the laboratory, but our most recently funded USDA/AFRI project focused on amphotericin B as an innate immune system adjuvant in cattle will form the core of the renewal. We plan on submitting at least two major research grants in support of this work in the coming year.

Impacts
What was accomplished under these goals? The direct impact of this work is on our general understanding of the bovine immune system and how it is similar and dissimilar to the immune system in rodents and humans. This information should facilitate development of new approaches to treat infectious diseases of cattle. Hatch and competitive grant funding, as well as research facilities provided by MAES, led to the specified outputs. As in previous years, this project continues to fund the general operation of our laboratory and supports our hatch project on bovine gamma/delta T cells. 2014 represented another productive year for our project. We continued our analyses of new adjuvant materials that activate bovine gamma/delta T cells in vivo, including in bovine calves. One new adjuvant is an approved anti-fungal drug in the clinical and we have shown is a potent adjuvant for bovine gamma/delta T cells. Work on an AFRI/USDA grant was on this new adjuvant material was started in December, 2013, and the first paper from this work was submitted and is under review. Our work on nutritional supplements, such as those rich in polyphenols, progressed as well, and we have identified an approach to mitigate colitis which appears to be due to a unique effect on T cells. Finally, our work has continued in multiple infection disease models including diseases caused by Salmonella sps, Coxiella burnetii, and influenza. A new effort on Clostridium difficile was started in 2013/2014. Per the latter, we have found unexpected results concerning the role for type I IFNs, which will be tested further in confirmation experiments in the coming year.

Publications

  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Snyder, D.T., Robison, A., Kemoli, S., Kimmel, E., Jutila, M.A., and Hedges, J.F. 2014. Oral delivery of oligomeric procyanidins in Applepoly� enhances type I IFN responses in vivo. J. Leuk. Biol., 94:841-847.PMCID:PMC3984970.
  • Type: Journal Articles Status: Submitted Year Published: 2014 Citation: Ramstead, A.G., Schepetkin,I.A., Todd, K., Loeffelholz, J., Berardinell, J.G., Quinn, M.T., and Jutila, M.A. 2014. Ageing influences the response of T cells to simulation by the ellagitannin, oenothein B. Submitted.
  • Type: Journal Articles Status: Under Review Year Published: 2014 Citation: Hedges, J.F., Mitchell, A.M., Jones, K., Kimmel,E., Ramstead, A., Snyder, D., and Jutila, M.A. 2014. AmB stimulates gd T cells and NK cells and enhancesprotection from Salmonella infection. Submitted.


Progress 01/01/13 to 09/30/13

Outputs
Target Audience: Researchers in the field of bovine immunology. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? One PhD and one MS student worked on this project in the past year. How have the results been disseminated to communities of interest? Yes, to the sceintific community through publications and meeting presentations. What do you plan to do during the next reporting period to accomplish the goals? We will continue as we have done in previous years. We have no plans for any major changes in our program. We plan to submitt at least two grants for additional funding in the coming year.

Impacts
What was accomplished under these goals? The direct impact of this work is on our general understanding of the bovine immune system and how it is similar and dissimilar to the immune system in rodents and humans. This information should facilitate development of new approaches to treat infectious diseases of cattle. Hatch and competitive grant funding, as well as research facilities provided by MAES, led to the specified outputs. As in previous years, this project continues to fund the general operation of our laboratory and supports our hatch project on bovine gamma/delta T cells. 2013 represented another productive year for our project. We continued our analyses of new adjuvant materials that activate bovine gamma/delta T cells in vivo, including in bovine calves. One new adjuvant is an approved anti-fungal drug in the clinical and we have shown is a potent adjuvant for bovine gamma/delta T cells. An AFRI/USDA grant was submitted on this new adjuvant material and it was funded in December 2013. Our work on nutritional supplements, such as those rich in polyphenols, progressed as well, and we have identified an approach to mitigate colitis which appears to be due to a unique effect on T cells. Finally, our work has continued in multiple infection disease models including diseases caused by Salmonella sps, Coxiella burnetii, and influenza. A new effort on Clostridium difficile was started in 2013.

Publications

  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Kouakou, K., Scheptekin, I.A., Yapi, A., Kirpotina, L.N., Jutila, M.A., and Quinn, M.T. 2013. Immunomodulatory activity of polysaccharides isolated from Alchornea cordifolia. J. Ethnopharm., 7;146(1):232-242.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Hedges, J.F, Snyder, D.T., Jerome, M., Kimmel, E.M., Holderness, J., and Jutila, M.A. 2013. SLC11A1 is expressed by innate lymphocytes and augments their activation. J. Immunol. 190:4263.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Schepetkin, I.A., Kouakou, K., Yapi, A., Kirpotina, L.N., Jutila, M.A. and Quinn, M.T. 2013. Immunomodulation and hemagglutination activity of acidic polysaccharides isolated from Combretum recemosum. Int Immunopharmacol. 15(3):628-637.
  • Type: Book Chapters Status: Published Year Published: 2013 Citation: Holderness, J, Hedges, J.F., Ramstead, A., and Jutila, M.A. 2013. Comparative biology of yo T cell function in humans, mice and domestic animals. Annual Review Animal Biosciences, Volume 1, pg 99-124..
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Kouakou, K., Schepetkin, I.A., Jun, S., Kirpotina, L.N., Yapi, A., Pascual, D.W., Jutila, M.A., and Quinn, M.T. 2013. Immunomodulatory activity of polysaccharides isolated from Clerodendrum splendens; Beneficial effects in experimental autoimmune encephalomyelitis. BMC Complement Altern Med. 13:149.


Progress 01/01/12 to 12/31/12

Outputs
OUTPUTS: As in previous years, this project continues to fund the general operation of our laboratory and supports our hatch project on bovine gamma/delta T cells. 2012 represented another productive year for our project. We continued our analyses of new adjuvant materials that activate bovine gamma/delta T cells in vivo, including in bovine calves. One new adjuvant is an approved anti-fungal drug in the clinic, which we hope to develop as a novel immunomodulatory agent in calves. An AFRI/USDA grant was submitted on this new adjuvant material. In the past year we have shown that NRAMP-1 augments gamma/delta T cell activation and subsequent production of IFN-gamma in cattle, humans and mice and have just recent had a paper describing these results accepted in the Journal of Immunology. Our work on nutritional supplements, such as those rich in polyphenols, progressed as well, and we have identified a unique approach to mitigate neutropenia which appears to be due to enhanced CSF production and responses to type I IFNs. Finally, our work has continued in multiple infection disease models including diseases caused by Salmonella sps, Coxiella burnetii, Brucella sps, Francisella tularensis and influenza virus. PARTICIPANTS: Mark Jutila served as PI on this project. Amanda Robison served as a part-time research associate. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
The direct impact of this work is on our general understanding of the bovine immune system and how it is similar and dissimilar to the immune system in rodents and humans. This information should facilitate development of new approaches to treat infectious diseases of cattle. Hatch and competitive grant funding, as well as research facilities provided by MAES, lead to the specified outputs, which are predominantly peer-reviewed publications and meeting presentations. In the current year (2013), we submitted a new grant to AFRI based on some of the adjuvant work described above.

Publications

  • Khlebnikov, A.I., Schepetkin, I.A., Kirpotina L.N., Brive, L., Dahlgren, C., Jutila, M.A., and Quinn, M.T. 2012. Molecular docking of 2-(bensimidazol-2-ylthio)-N-phenylacetamide-derived small-molecule agonists of human formyl peptidereceptor 1. J. Mol. Mod., 18:2831.
  • Ramstead, A.G., Schepetkin, I.A., Quinn, M.T., Jutila, M.A. 2012. Oenothein B, a cyclic dimeric ellagitannin isolated from Epilobium angustifolium, enhances IFN-g production by lymphocyes. PLoS One. 7:e50546.
  • Schepetkin, I.A., Kirpotina, L.N., Khlebnikov, A.I., Hanks, T.S., Kochetkova, I., Pascual, D.W., Jutila, M.A. and Quinn, M.T. 2012. Identification and characterization of a novel class of c-Jun N-terminal kinase inhibitors. Mol. Pharmacol. 81:832.
  • Ramstead, A and Jutila, M.A. 2012. Complex role of γδ T cell-derived cytokines and growth factors in cancer. J. Interferon Cytokine Res. 32: 563.
  • Kouakou, K., Scheptekin, I.A., Yapi, A., Kirpotina, L.N., Jutila, M.A., and Quinn, M.T. 2013. Immunomodulatory activity of polysaccharides isolated from Alchornea cordifolia. J. Ethnopharm., in press.
  • Hedges, J.F, Snyder, D.T., Jerome, M., Kimmel, E.M., Holderness, J., and Jutila, M.A. 2013. SLC11A1 is expressed by innate lymphocytes and augments their activation. J. Immunol. In press.


Progress 01/01/11 to 12/31/11

Outputs
OUTPUTS: As in previous years, this project continues to fund the general operation of our laboratory and supports our hatch project on bovine gamma/delta T cells. 2011 represented one of the most productive years for our project. We continued our analyses of new adjuvant materials that activate bovine gamma/delta T cells in vivo, including in bovine calves. One new adjuvant is an approved anti-fungal drug in the clinic, which we hope to develop as a novel immunomodulatory agent in calves. We completed an NRI funded study in 2011 where we have defined the selective expression of NRAMP-1 in bovine gamma/delta T cells. In the past year we have shown the NRAMP-1 augments gamma/delta T cell activation and subsequent production of IFN-gamma. We continued work in the area of nanotechnology, specifically using nanoparticles to deliver novel adjuvant materials to the lung. Our work on nutritional supplements progressed as well, and we have identified a unique approach to mitigate neutropenia. Finally, our work has continued in multiple infection disease models including diseases caused by Salmonella sps, Coxiella burnetii, Brucella sps, Francisella tularensis and influenza virus. PARTICIPANTS: Mark Jutila served as PI on this project. Amanda Robison served as a part-time research associate. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
The direct impact of this work is on our general understanding of the bovine immune system and how it is similar and dissimilar to the immune system in rodents and humans. This information should facilitate development of new approaches to treat infectious diseases of cattle. Hatch and competitive grant funding, as well as research facilities provided by MAES, led to the specified outputs, which are predominantly peer-reviewed publications and meeting presentations. In the coming year, we plan on submitting a new grant to AFRI based on some of the adjuvant work described above.

Publications

  • Kimmel, E., Jerome, M., Holderness, J., Snyder, D., Kemoli, S., Jutila, M.A., and Hedges, J.F. 2011. Oligomeric procyanidins stimulate innate antiviral immunity in Dengue virus infected human PBMCs. J. Anti-viral Res., 90(1):80-86.
  • Holderness, J., Schepetkin, I.A., Freedman, B., Kirpotina L.N., Quinn, M. T., Hedges, J.F., and Jutila, M.A. 2011. Polysaccharides isolated from Acai fruit induce innate immune responses. PLoS One, 6(2):e17301.
  • Skyberg, J.A., Robison, A., Golden, S., Rollins, M.F., Callis, G., Huarte, E., Kocvhetkova, I., Jutila, M.A., and Pascual, D.W. 2011. Apple polyphenols require T cells to ameliorate dextran sulfate sodium-induced colitis and dampen proinflammatory cytokine expression. J. Leuk. Biol., (6):1043-1054.
  • Skyberg, J.A., Thornburg, T., Rollins, M.R., Huarte, E., Jutila, M.A., and Pascual, D.W. 2011. Murine and bovine γδ T cells enhance innate immunity against Brucella abortus infections. PLoS One 6: (7):e21978
  • Khlebnikov, A.I., Schepetkin, I.A., Kirpotina L.N., Brive, L., Dahlgren, C., Jutila, M.A., and Quinn, M.T. 2011. Molecular docking of 2-(bensimidazol-2-ylthio)-N-phenylacetamide-derviced small-molecule agonists of human formyl peptide receptor 1. J. Mol. Mod., in press.
  • Skyberg, J.A., Rollins, M.F., Holderness, J., Marlenee, N., Scheptekin, I., Goodyear, A., Dow, S., Jutila, M.A., and Pascual, D.W. 2012. Nasal Acai polysaccharides potentiate innate immunity to protect against pulmonary Francisella tularensis and Burkholderia pseudomallei infections. PloS Pathogens, in press.
  • Shipman, M., Lubick, K., Focuhard, D., Guram, R., Grieco, P., Jutila, M.A., and Dratz, E.A. 2011. Proteomic and systems biology analysis of monocytes exposed to securinine, a GABAA receptor antagonists and immune adjuvant. Submitted, minor comments to address.


Progress 01/01/10 to 12/31/10

Outputs
OUTPUTS: This project continues to fund the general operation of our laboratory and supports our hatch project on bovine gamma/delta T cells. We continue to make progress in all aspects of this work. We have further defined new adjuvant materials that activate bovine gamma/delta T cells in vivo, including in bovine calves. One new adjuvant is an approved anti-fungal drug in the clinic, which we are planning on testing in two different infectious diseases settings in cattle. Our gene expression work continues to progress well. Last year we identified a novel immunoregulatory cytokine produced by gamma/delta T cells, gas-6. Studies continue on its function within infectious disease and inflammatory conditions. In our NRI funded study, we have further defined the selective expression of NRAMP-1 in bovine gamma/delta T cells. We continued work in the area of nanotechnology, specifically using nanoparticles to deliver novel adjuvant materials to the lung. Finally, our work has continued in multiple infection disease models including diseases caused by Salmonella sps, Coxiella burnetii and dengue virus. PARTICIPANTS: Mark Jutila served as PI on this project. Amanda Robison served as a part-time research assistant. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project. PARTICIPANTS: Mark Jutila served as project director in the past year. Amanda Robison continued to serve as a part time research associate on the project. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
The direct impact of this work is on our general understanding of the bovine immune system and how it is similar and dissimilar to the immune system in rodents and humans. This information should facilitate development of new approaches to treat infectious diseases of cattle. Data acquired from our gene expression studies led to the development of a new NRI funded project in the past two years. In the coming year, we plan on submitting a new grant to AFRI based on some of the adjuvant work described above.

Publications

  • Kirpotina, L.N., Khlebnikov, A.I., Schepetkin, I.A., Ye, R.D., Rabiet, M-J, Jutila, M.A., and Quinn, M.T. 2010. Identification of novel small-molecule agonists for human formyl peptide receptors and pharmacophore models of their recognition. Mol. Pharm., 77:159.
  • Daughenbaugh, K., Holderness, J., Graff, J.C., Hedges, J.F., Freedman, B., Graff, J.W., and Jutila, M.A. 2011. Contribution of transcript stability to a conserved procyanidin-induced cytokine response in γδ T cells. Genes and Immunity, in press.
  • Schepetkin. I., Kirpotina, L.N., Khlebnikov, A.I., Jutila, M.A., and Quinn, M.T. 2011. Gastrin-releasing peptide/neuromedin B receptor antagonists PD176252, PD168368, and related analogs are potent agonists of human formyl-peptide receptors. Mol. Pharm. Jan;79(1):77-90.


Progress 01/01/09 to 12/31/09

Outputs
OUTPUTS: This project continues to fund the general operation of our laboratory and supports our hatch project on bovine gamma/delta T cells. We have continued to make progress in all aspects of this work. We have continued testing of new adjuvant materials that activate bovine gd T cells in vivo, including in bovine calves. Our gene expression work continues to progress well. We have identified a novel immunoregulatory cytokine produced by gd T cells, gas-6. Studies are underway on defining its function within infectious disease and inflammatory conditions. We continued work in the area of nanotechnology, specifically using nanoparticles to deliver novel adjuvant materials to the lung. Finally, our work has continued in multiple infection disease models including diseases caused by Salmonella sps, Coxiella burnetii and dengue virus. PARTICIPANTS: Mark Jutila served as PI on this project. Amanda Robison served as a part-time research assistant. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
The direct impact of this work is on our general understanding of the bovine immune system and how it is similar and dissimilar to the immune system in rodents and humans. This information should facilitate development of new approaches to treat infectious diseases of cattle. Data acquired from our gene expression studies led to the development of a new NRI funded project in the past year. In the coming year, new results on gas-6 will be translated into a new project. The main evaluation of this project was reflected in 4 peer-reviewed publications in press in the past year--see below.

Publications

  • Graff, J.C, Kimmel, E.M., Freedman, B.Schepetkin, I.A., Holderness, J., Quinn, M.T., Jutila, M.A. and Hedges, J.F. 2009. Polysaccharides derived from Yamoa (Funtumia elastica) prime gammadelta T cells in vitro and enhance innate immune responses in vivo. Int. Immunopharmacol, 9:1313-22.
  • Siemsen, D.W., Kirpotina, L.N., Jutila, M.A., and Quinn, M.T. 2009. Inhibition of the human neutrophil NADPH oxidase by Coxiella burnetii. Microbes Infect., 6-7:671-679.
  • Wiley, J.A, Richert, L.E., Swain, S.D. Harmsen, A., Barnard, D. L., Randall, T. D., Jutila, M., Douglas, T., Broomell, C., Young, M., and Harmsen, A. 2009. Inducible Bronchus-associated lymphoid tissue elicited by a protein cage nanoparticle enhances protection in mice against diverse respiratory viruses. PLoS One, 4: e7142.
  • Schepetkin. I., Kirpotina, L.N., Khlebnikov, A.I., blaskovich, C.L., Jackiw, L., Jutila, M.A., and Quinn, M.T. 2009. Immunomodulatory Activity of Oenothein B Isolated from Epilobium angustifolium. J. Immunol., 183:6754-66.


Progress 01/01/08 to 12/31/08

Outputs
OUTPUTS: This project funds the basic operation of our laboratory and supports our hatch project on bovine gamma/delta T cells. We have continued to make progress in all aspects of this project. We have initiated testing of new adjuvant materials that activate bovine gd T cells in vivo, including in bovine calves. Our gene expression work continues to progress well. We were able to adapt RNA-interference technologies to the study of NOD receptors in bovine gd T cells. We continued work in the area of nanotechnology, specifically using nanoparticles to deliver novel adjuvant materials to the lung. Finally, our work has expanded into multiple infection disease models including diseases caused by Salmonella sps, Coxiella burnetii and dengue virus. PARTICIPANTS: Mark Jutila served as PI Jill Graff served as a part-time research associate PARTICIPANTS: Not relevant to this project. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
The direct impact of this work is on our general understanding of the bovine immune system and how it is similar and dissimilar to the immune system in rodents and humans. This information should facilitate development of new approaches to treat infectious diseases of cattle. We have identified new adjuvant materials that enhance the activity of the bovine gd T cells. These materials may be useful in enhancing the general health of calves directly or they might be used in enhancing vaccine responses.

Publications

  • Schepetkin, I.A., Xie, G., Jutila, M.A., and Quinn, M.T. (2008) Complement-fixing Activity of Humic Substances from Shilajit and Other Natural Sources. Phytother. Res., In Press.
  • Kerns, M.M., Jutila, M.A., and Hedges, J.F. 2009. The distinct responses of γδ T cells to the Nod2 agonist, muramyl dipeptide. Cellular Immunol in press.
  • Nemali S., Siemsen, D.W., Nelson, L.K., Bunger, P.L., Faulkner, C.L., Rainard, P., Gauss, K.A., Jutila, M.A., Quinn MT. 2008. Molecular analysis of the bovine anaphylatoxin C5a receptor. J. Leukoc Biol. 84:537-549.
  • Schepetkin, I.A., Xie, G., Kirpotina, L.N., Jutila, M.A., and Quinn, M.T. (2008) Macrophage immunomodulatory activity of polysaccharides isolated from Opuntia polyacantha. Int. Immunopharmacol., In press.
  • Holderness, J., Hedges, J., Daughenbaugh, K., Kimmel, E., Graff, J., Freedman, B., and Jutila, M.A. 2008. Response of gamma/delta T cells to plant-derived tannins. Critical Rev. Immunol., 28: 377-402.


Progress 01/01/07 to 12/31/07

Outputs
OUTPUTS: This project funds the basic operation of our laboratory and supports our hatch project on bovine gamma/delta T cells. We have continued to make progress in all aspects of this project. In the past year, we completed our high throughput screening efforts for gamma/delta T cell agonists that could be used as novel "drugs" to enhance innate immunity. We have initiated testing of novel agonists in in vivo models, including bovine calves. Our gene expression work continues to progress well. We were able to adapt RNA-interference technologies to the study of NOD receptors in bovine gd T cells. We continued work in the area of nanotechnology, specifically using nanopraticles to deliver novel adjuvants. We also established a new collaboration in which we help define the functional properties of a new inhibitor of T cell function. PARTICIPANTS: Mark Jutila served as PI Jill Graff served as a part-time research associate

Impacts
The direct impact of this work is on our general understanding of the bovine immune system and how it is similar and dissimilar to the immune system in rodents and humans. This information should facilitate development of new approaches to treat infectious diseases of cattle.

Publications

  • Ren, Y., Strobel, G., Graff, J., Jutila, M., Park, S.G., Gosh, S., Teplow, D., Condron, M., Pang, E., and Hess, W.M. 2008. Volutellin A, an immunosuppressive peptide from Volutella sps, Microbiology, in press.
  • Jutila, M.A., Holderness, J., Graff, J. and Hedges, J.F. 2008. Antigen independent priming: A transitional response of bovine gd T cells to infection. AHRP, in press.
  • Hedges, J.F., Buckner, D., Rask, K., Kerns, H., Jackwi, L. Theresa Trunkle, David Pascual and Jutila, M.A. 2007. Mucosal lymphatic-derived gd T cells respond early to experimental Salmonella enterocolitis by increasing expression of IL-2Ra. Cell. Immunol., 246: 8.
  • Lubick, K., Radke, M., and Jutila, M.A. 2007. Securinine, a GABAA receptor antagonist, enhances macrophage clearance of phase II C. burnetii: comparison to TLR agonists. J. Leuk. Biol. 82:1062.
  • Graff, J. and Jutila, M.A. 2007. Differential regulation of CD11b on gamma/delta T cells and monocytes in response to unripe apple polyphenols. J. Leuk. Biol., 82: 603.
  • Holderness, J., Jackwi, L., Kimmel, E., Kerns, H., Radke, M., Hedges, J., Petrie, C., McCurley, P., Glee, P., Palecanda, A., and Jutila, M.A. 2007. Select plant polyphenols (condensed tannins) enhance IL-2R upregulation and cell division in gd T cells. J. Immunol., 179: 6468.


Progress 01/01/06 to 12/31/06

Outputs
We have continued to make progress in all aspects of this project. In the past year, we have continued our high throughput screening efforts for gamma/delta T cell agonists that could be used as novel "drugs" to enhance innate immunity. We have screened >150,000 "drug-like" candidates in three different functional assays. Our gene expression work has progress well. Specifically, we completed our analyses of global gene expression in mucosal alpha/beta versus gamma/delta T cells. We extended our analysis of innate receptors on gamma/delta T cells by showing that these cells express NOD2 and respond to the NOD2 agonist, muramyl dipeptide. New to the past year was the establishment of new collaborative projects in the area of nanotechnology. These new collaborations have already led to two publications.

Impacts
The direct impact of this work is on our general understanding of the bovine immune system and how it is similar and dissimilar to the immune system in rodents and humans. This information should facilitate development of new approaches to treat infectious diseases of cattle

Publications

  • Uchida, M., Flenniken, M., Allen, M., Willits, D.A., Crowley, B.E., Brumfield, S., Willis, A.F., Jackiw-Turk, L., Jutila, M., Young, M. and Douglas, T. 2006. Cancer Cell Targeting Ferrimagnetic Iron Oxide Nanoparticle Incorporated into a Ferritin Cage Architecture. J. Amer. Chem. Soc.,128:16626.
  • Kaiser, C.R., Flenniken, M., Harmsen, A.G., Harmsen, A. L., Jutila, M.A., Douglas, T.,and Young, M.J. 2006. Biodistribution studies of protein cage nanoparticles demonstrate broad tissue distribution and rapid clearance in vivo. Intern. J. Nanomedicine, in press.
  • Hedges, J.F., Kerns, M.M., and Jutila, M.A. 2006. The distinct responses of gd T cells to the Nod2 agonist, muramyl dipeptide. Submitted


Progress 01/01/05 to 12/31/05

Outputs
We have continued to make progress in all aspects of this project. In the past year, we have continued our high throughput screening efforts for gamma/delta T cell agonists that could be used as novel "drugs" to enhance innate immunity. We have screened 10s of thousands of "drug-like" candidates in three different functional assays. Our gene expression work in gamma/delta T cells continue to progress well. We defined a novel pattern of gene expression induced by LPS treatment of these cells, suggesting that LPS and other PAMPs "prime" gamma/delta T cells leading to more robust reponses against downstream agonists. We continued our use of the Affymetrix Oligonucleotide array system to study global gene expression in bovine lymphatic T cells prior to and during the course of Salmonella infection. We have compared responses of alpha/beta and gamma/delta T cells. We published our comprehensive SAGE analysis of bovine blood and spleen gamma/delta T cell subset and alpha/beta T cells in their resting state and following mitogen stimulation. We have also defined global gene expression human gamma/delta T cell subsets. We have continued to make progress in our analysis of innate immune system receptors expressed by gamma/delta T cells. Finally, a number of different collaborations have been maintained or newly established, which has expanded our work on gamma/delta T cells and other cell types, such as bovine epithelial cells.

Impacts
The direct impact of this work is on our general understanding of the bovine immune system and how it is similar and dissimilar to the immune system in rodents and humans. This information should facilitate development of new approaches to treat infectious diseases of cattle

Publications

  • Lahmers, K.K., Hedges, J.F., Jutila, M.A., Deng, M., Abrahamsen, M.S., and Brown, W.C. 2005. Differences in immune response gene expression by WC1+ gamma/delta and CD4+ alpha/beta T cell clones that respond to Anaplasma marginale major surface protein 2 indicate functional heterogeneity between these T lymphocyte subsets. Submitted.
  • Hedges, JF, KM Rask, DL Buckner, LO Jackiw, H Willmore, T Trunkle, M Deng, MS Abrahamsen, and MA Jutila. 2005. Genomic Analysis in T cell subsets following experimental Salmonella Enterica Serovar Typhimurium (S. typhimurium) infection in the calf. XIII International Conference on the Status of Plant and Animal Genome Research. San Diego, CA. P695.
  • Abrahamsen, MS, WC Brown, and MA Jutila. 2005. Functional genomics of bovine gamma/delta T cells. XIII International Conference on the Status of Plant and Animal Genome Research. San Diego, CA. W056.
  • Graff, JC, MA Jutila. 2005. Identification of functional B lymphocyte induced maturation protein-1 (BLIMP-1) in gd T cells. FASEB J. (Abstract) 46.33.
  • Hedges, JF, KM Rask, DL Buckner, LO Jackiw, H Willmore, T Trunkle, M Deng, MS Abrahamsen, and MA Jutila. 2005. Genomic Analysis in T cell subsets following experimental Salmonella Enterica Serovar Typhimurium (S. typhimurium) infection in the calf. FASEB J. (Abstract) 261.44.
  • Kress, ER, J Hedges, and M Jutila. 2005. Expansion/activation of the Vdelta1 subset of human gamma delta T cells in vitro. FASEB J. (Abstract) 615.8.
  • Shaneyfelt, M., Burke, A.D., Jutila, M.A., and Hardy, M.E. 2005. Natural products that reduce rotavirus infectivity identified by a cell-based semi-high-throughput screening assay. Submitted.
  • Buckner, D., Wilson, S., Kurk, s., Hardy, M., Miessner, N. and Jutila, M.A. 2005. Use of early passage fetal intestinal epithelial cells in high throughput screening assays: an approach to identify new innate immune system adjuvants. J. Biomolecular Screen, in press.
  • Graff, JC, M Behnke, J Radke, M White, D Lee and MA Jutila. 2005 A comprehensive SAGE database and bioinformatics tool resource for the analysis of bovine gd T cells. XIII International Conference on the Status of Plant and Animal Genome Research. San Diego, CA. P838.
  • Hedges, JF, and MA Jutila. 2005. Response of gammadelta T cells to the NOD2 agonist muramyl dipeptide. FASEB J. (Abstract) 826.15.
  • Lubick, K, and Jutila M. 2005. Expression of CD36 on bovine gamma/delta T cells. FASEB J. (Abstract) 859.16.
  • Holderness, J, D Buckner, K Rask, M Radke, N Crowly, S Kurk, S Hogan, K Day, JF Hedges, and MA Jutila. 2005. Stimulation of bovine gd T cells with agonists including tea extracts, and bacterial, viral and chemical compounds. FASEB J. Abstract LB608.
  • Hedges J. F., Lubick, K., and Jutila, M.A. 2005. gd T cells respond directly to pathogen-associated microbial products. J. Immunology 174: 6045.
  • Kress, E., Hedges, J.F., and Jutila, M.A. 2005. Gene expression in in vitro expanded human Vdelta 1 and V delta 2 gamma/delta T cells following treatment with endotoxin or phorbol ester. Molecular Immunology, in press.
  • Graff J. Behnke, M., Radke, J., White, M. and Jutila, M.A. 2005. A Comprehensive SAGE Database for the Analysis of gamma/delta T Cells. International Immunology, in press.
  • Lubick, K and Jutila, M.A.. 2005. LTA Recognition by Bovine gamma/delta T cells Involves CD36. Journal of Leukocyte Biology, in press.
  • Hedges, J., Buckner, D.L., Rask, K.M., Jckwi, L.O. Thrunckle, T., Pascual, D.W., and Jutila, M.A. 2005. Genomic analysis of mucosal derived ab and gamma/delta T cells during experimental Salmonella enterica serovar typhimurium enterocolitis. Submitted.


Progress 01/01/04 to 12/31/04

Outputs
We have continued to make progress in all aspects of this project. In the past year, we have initiated a high throughput screening effort for gamma/delta T cell agonists that could be used as novel "drugs" to enhance innate immunity. We have screened nearlly 100,000 "drug-like" candidates. Our gene expression work in gamma/delta T cells have progressed well and we defined a novel pattern of gene expression induced by LPS treatment of these cells. We adapted the use of the Affymetrix Oligonucleotide array system to study global gene expression in bovine lymphatic T cells prior to and during the course of Salmonella infection. We have compared responses by alpha/beta and gamma/delta T cells. We characterized the impact of poly unstaturated fatty acids on the regulation of bovine leukocyte antigen expression. We defined a unique effect on L-selectin with respect to regulating linkage to the cytoskelton.

Impacts
The direct impact of this work is on our general understanding of the bovine immune system and how it is similar and dissimilar to the immune system in rodents and humans. This information should facilitate development of new approaches to treat infectious diseases of cattle.

Publications

  • Leid, J. and Jutila, M.A. 2004. Impact of Polyunsaturated Fatty Acids on Cytoskeletal linkage of L-selectin. Cellular Immunol, 228: 91-98.
  • Hedges, J.F., Lubick, K., and Jutila, M.A. 2004. gd T cells respond directly to pathogen associated molecular patterns. J.Immunol. in press.
  • Graff, J.C., Behnke, M., Radke, J., White, M., Lee, D., and Jutila, M.A. 2004. A comprehensive SAGE database for the analysis of gd T cells. Submitted.
  • Lancto, C.A., Walcheck, B., Jutila, M.A., and Abrahamsen, M.S. 2004 Role of PNAd in lymphocyte recruitment during Cryptosporidium parvum infection. Submitted.
  • Graff J.C., Hedges, J.F., Behnke, M., Radke, J., White, M., and Jutila, M.A. 2004. Analysis of gene expression in gamma/delta T cells. Plant and Animal Genome, XII, P647.
  • Graff J.C., Hedges, J.F., and Jutila, M.A. 2004. B lymphocyte-induced maturation protein-1 (Blimp-1) in gamma/delta T cells. FASEB, LB186.
  • Jutila, M., Graff, J., and Hedges, J.F. 2004. Gene Expression in bovine gd T cells. 7th International Veterinary Immunology Symposium, WK.8.6.6


Progress 01/01/03 to 12/31/03

Outputs
This Hatch project covers all aspects of our research on bovine gamma/delta T cells. Overall, we have succeeded in many different projects related to our work on these bovine cells. For example, we have characterized chemokines and adhesion molecules that regulate the trafficking of CD8- versus CD8+ gamma/delta T cells. We have made considerable progress in our gene expression analysis of bovine blood and spleen gamma/delta T cells and have compared gene expression profiles in tissue-specific subsets, and resting and activating cells. This work has led to direct evidence for a functional link between gamma/delta T cells and myeloid cells. Our new studies on the role of bovine gamma/delta T cells in the host response against viral, bacterial and protozoal mucosal pathogens have progressed well and we currently are focussing our efforts in crytptosporidiosis and salmonellosis. Based, in part, on our work on bovine chemokines, we have intiated a new NIH funded project on the identification of new "adjuvants" that act on gamma/delta T cells, enhancing their function within the innate immune system.

Impacts
The direct impact of this work is on our general understanding of the bovine immune system and how it is similar and dissimilar to the immune system in rodents and humans. This information should facilitate development of new approaches to treat infectious diseases of cattle.

Publications

  • Hedges, J.F., Graf, J, and Jutila, M.A. 2003. Transcriptional profiling of gamma/delta T cells. J. Immunol. 171: 4960.
  • Daughenbaugh, J., Hedges, J.F., Behnke M., Radke J., White M., and Jutila, M.A. 2003. Use Of Serial Analysis of Gene Expression (SAGE) to analyze gene expression profiles in resting and activated bovine spleen ab and gd T cells. FASEB J. 17: 29.10
  • Hedges, J.F. and Jutila, M.A. 2003. Gd T cells respond directly to pathogen-associated microbial products. FASEB J. 17: 79.28


Progress 01/01/02 to 12/31/02

Outputs
This Hatch project covers all aspects of our research on bovine gamma/delta T cells. We have succeeded in characterizing chemokines and adhesion molecules that regulate the trafficking of CD8+ gamma/delta T cells to the gut mucosa. We have made considerable progress in our gene expression analysis of bovine blood and spleen gamma/delta T cells and have compared gene expression profiles in tissue-specific subsets and resting and activating cells. This work has led to direct evidence for a functional link between gamma/delta T cells and myeloid cells. We have also studied the expression of L-selectin and other important immune system molecules on bovine and human gamma/delta T cells. Finally, studies of bovine rotavirus, salmonellosis, and cryptosporidiosis have been initiated.

Impacts
The direct impact of this work is on our general understanding of the bovine immune system and how it is similar and dissimilar to the immune system in rodents and humans. This information should facilitate development of new approaches to treat infectious diseases of cattle.

Publications

  • Csencsits, K.L., Jutila, M.A., and Pascual, D.W. 2002. Mucosal addressin (MAdCAM-1) expression and naive lymphocyte binding interactions vary among the cranial, oral, and nasal-associated lymphoid tissues (CONALT). Eur. J. Immunol., 32:3029.
  • Jutila, M.A., Kurk S., Jackiw L., Knibbs R.N., and Stoolman, L.M. 2002. L-selectin can serve as an E-selectin ligand on human T-cell blasts. J. Immunol. 169: 1768.
  • Wilson, E., Hedges J. F., Butcher E.C., Briskin M., and Jutila, M.A. 2002. Bovine gd T cell subsets express distinct patterns of chemokine responsiveness and adhesion molecules. A mechanism for tissue specific gd T cell subset accumulation. J. Immunol., 169: 4970.
  • Meissner N., Radke J., Hedges J. F. White M., Behnke M., Bertolino S., Abrahamsen M. Hedges, and Jutila, M.A. 2003. Serial analysis of gene expression (SAGE) in circulating gd T cells . J. Immunol., 170: 356.
  • Hedges, J. F., Cockrell, D., Jackiw, L., Meissner, N., and Jutila, M.A. 2003. Differential mRNA expression in circulating gd T lymphocyte subsets defines unique tissue-specific functions. J. Leuk. Biol., 73: 306.
  • Meissner, N., Radke, J., Jackiw, L., Behnke, M., White, M., Abrahamsen, M., Hedges, J., and Jutila, M.A. 2002. Functional genomic analysis of bovine CD8+ versus CD8- gamma/delta T cells. FASEB J. 16:D248.20.
  • Hedges, J., Cockrell, D., Jackiw, L., Meissner, N., and Jutila, M.A. 2002. Array analysis of differential mRNA expression by subsets of bovine gd T cells. FASEB J. 16:D248.19.
  • Pascual, D.W., Trunkle, T., Callis, G., and Jutila, M.A. 2002. Attenuation of Salmonella-induced inflammation: fimbriae reduce proinflammatory cytokines and prevent neutrophil influx into the Peyer's patches. FASEB J. 16:D236.
  • Kurk, S., Jackiw, L., Willmore, H., Knibbs, R., Stoolman, L., and Jutila, M.A. 2002. Role of L-selectin as an E-selectin ligand on human T cells: comparison to PSGL-1 and other E-selectin binding glycoproteins. FASEB J. A924.2.


Progress 10/01/00 to 09/30/01

Outputs
This project provides funds for salary in support of our overall studies on bovine gamma/delta T cells. Our experimental aims relating specifically to the Hatch project deal with the analysis of a number of new gamma/delta T cell specific monoclonal antibodies we have recently generated.Using these antibodies we have defined tissue-specific subsets of gamma/delta T cells. Current studies are focused on the trafficking of these subsets and using the antibodies to purify the subsets for functional gene analysis. The reagents generated in this project have also been used in the study of bovine neutrophils and blue tongue virus infected bovine endothelial cells.

Impacts
It is anticipated that these results will lead to new adjuvants and vaccines that selectively target gamma/delta T cells, which will improve vaccination protocols for certain pathogens.

Publications

  • Leid, J.G., Speer, C.A., and Jutila, M.A. 2001. Ultrastructural Examination of Cytoskeletal Linkage of L-selectin and Comparison of L-selectin Cytoskeletal Association to that of Other Human and Bovine Lymphocyte Surface Antigens. Submitted.
  • Quinn, M.T., Swain S.D., Parkos C.A., Jutila K.L., Siemsen D.W., Kurk, S.L., Jesaitis A.J., and Jutila M.A. 2001. A carbohydrate neoepitope that is up-regulated on human mononuclear leukocytes by neuraminidase treatment or cellular activation. Immunology, 104: 185-197.
  • DeMaula, C.D., Leutenegger C.M., Jutila, M.A., and MacLachlan, N.J. 2001. Bluetongue virus-induced activation of primary bovine lung microvascular endothelial cells. Vet. Immunol. Immunopath. in press.


Progress 10/01/99 to 09/30/00

Outputs
This project provides funds for salary in support of our overall studies on bovine gamma/delta T cells. Our experimental aims relating specifically to the Hatch project deal with the analysis of a number of new gamma/delta T cell specific monoclonal antibodies we have recently generated.Using these antibodies we have defined tissue-specific subsets of gamma/delta T cells. Current studies are focused on the trafficking of these subsets and using the antibodies to purify the subsets for functional gene analysis. The reagents generated in this project have also been used in the study of bovine neutrophils and blue tongue virus infected bovine endothelial cells.

Impacts
Bovine gamma/delta T cells are the predominant T cell population in newborns, yet we know very little of their importance to the immunological health of the calf.Understanding the function of these cells will be important in the development of new vaccines and adjuvants that target their responses.

Publications

  • Sipes, K., H. Edens, M. Kehrli, H. Miettinen, J. Cutler, M.A. Jutila, and M.Quinn. 1999. Analysis of surface antigen expression and host defense function in leukocytes from calves with bovine leukocyte adhesion deficiency. Am. J. Vet. Res., 60:1255-1261.
  • Kantele, J.M., S. Kurk, and M.A. Jutila. 2000. Effects of continuous exposure to SDF-1 on T cell rolling and tight adhesion to monolayers of activated endothelial cells. J. Immunol., 164: 5035-5040.
  • DeMaula, C.D., M.A. Jutila, D.W. Wilson and MacLachlan, N.J. 2000. Infection kinetics, prostacyclin release, and cytokine-mediated modulation of the mechanism of cell death during bluetongue virus infection of cultured ovine and bovine... J. Gen. Virol. in press.
  • Carter YM, Thomas R, Bargatze R, Jutila M, Murry C, Allen MD. 2000. Intracoronary E- and L-selectin blockade attenuates myocardial neutrophil infiltration in cardiac ischemia/reperfusion injury. Curr Surg. 57:639.
  • Leid, J.G., Steeber, D.A., Tedder, T.F., and Jutila, M.A. 2000. Antibody binding to a conformation-dependent epitope induces L-selectin associate with the detergent resistant cytoskeleton. J. Immunol., in press.
  • Miessner, N., Wilson, E., Jackiv-Turk, L., and Jutila, M.A. 2000. Functional analysis of CD8+ and CD8- gd T cells. FASEB J. 14:B436.
  • Leid, J. and Jutila, M.A. 2000. Comparison of cytoskeletal association of L-selectin to that of other gd T cell surface antigens. FASEB J. 14:B235.
  • Wilson, E., Briskin, M., and Jutila, M.A. 2000. A TCR-defined subset of bovine gd T cells express high levels of functional b7 integrin and preferentially accumulates in sites expression MAdCAM-1: a mechanism for tissue specific gd T cell accumulation. FASEB J. 14:B249.


Progress 01/01/99 to 12/31/99

Outputs
This project provides funds for salary in support of our overall studies on bovine gamma/delta T cells. Our experimental aims relating specifically to the Hatch project deal with the analysis of a number of new gamma/delta T cell specific monoclonal antibodies we have recently generated. We previously defined antibodies that recognize WC1 and the gamma/delta T cell receptor (TCR) for antigen. We have characterized four distinct subsets of gamma/delta T cells using these anibodies. One of the subsets preferentially localizes to sites of inflammation, whereas another specifically accumulates in the spleen. We are currently studying the basis for the tissue-selective recruitment of these subsets.

Impacts
Gamma/delta T cells are the predominant lymphocyte population in newborn calves. A better understanding of the function of these cells may lead to the development of better adjuvants and vaccines for infectious diseases of livestock.

Publications

  • Wilson, E., Aydintug, K., and Jutila, M.A. 1999. A circulating gamma/delta T cell subset, which is found in large numbers in spleen, accumulates inefficiently at sites of inflammation: correlation with lack of E-selectin ligands and L-selectin. J. Immunol., 162:4914-4919.
  • Sipes, K., H. Edens, M. Kehrli, H. Miettinen, J. Cutler, M.A. Jutila, and M.Quinn. 1999. Analysis of surface antigen expression and host defense function in leukocytes from calves with bovine leukocyte adhesion deficiency: Comparison of heterozygous and homozygous animals. Am. J. Vet. Res., 60:1255-1261.
  • Csencsits, K.L., M.A. Jutila, and D.W. Pascual. 1999. Nasal-associated lymphoid tissue (NALT): peripheral node addressin predominates in naive lymphocyte adhesion to HEV in a mucosal site. J. Immunol., 163:1382-1389.
  • Kantele, J.M., S. Kurk, and M.A. Jutila. 1999. Effects of continuous exposure to SDF-1 on T cell rolling and tight adhesion to monolayers of activated endothelial cells. Submitted.
  • Soltys, J., S.D. Swain, K.M. Sipes, L.K. Nelson, M.A. Jutila, and M.T. Quinn. 1999. Isolation of bovine neutrophils with biomagnetic beads: Comparison with standard percoll density isolation methods. J. Immunol. Meth. 226:71-84.


Progress 01/01/98 to 12/31/98

Outputs
This project provides funds for salary in support of our overall studies on bovine gamma/delta T cells. Our experimental aims relating specifically to the Hatch project deal with the analysis of a number of new gamma/delta T cell specific monoclonal antibodies we have recently generated. We have defined antibodies that recognize WC1 and the gamma/delta T cell receptor (TCR) for antigen. We have characterized four distinct subsets of gamma/delta T cells using these antibodies. One of the subsets preferentially localizes to sites of inflammation, whereas another specifically accumulates in the spleen. We are cuurently studying the basis for the tissue-selective recruitment.

Impacts
(N/A)

Publications

  • Kahn, J., B. Walcheck, G.I Migaki, M. Jutila and T.K. Kishimoto. 1998. Calmodulin regulates L-selectin adhesion molecule expression and function through a protease dependent mechanism: Direct association of calmodulin with the cytoplasmic tail of L-selectin. Cell 92:809-818.
  • Leid, J.G., C.A. Speer and M.A. Jutila. 1998. Comparative ultrastructure and expression of L-selectin on bovine alpha/beta and gamma/delta T cells. J. Leuk. Biol. 64:104-107.
  • Wilson, E., B. Walcheck, W. Davis, and M.A. Jutila. 1998. Preferential tissue localization of bovine gamma/delta T cell subsets defined by anti-T-cell receptor for antigen antibodies. Immunol. L. 64:39-44.


Progress 01/01/97 to 12/31/97

Outputs
This project provides funds for salary in support of our overall studies on bovine gamma/delta T cells. Our experimental aims relating specifically to the Hatch project deal with the analysis of a number of new gamma/delta T cell specific monoclonal antibodies we have recently generated. We have defined antibodies that recognize WC1 and the gamma/delta T cell receptor (TCR) for antigen. These antibodies have allowed the characteriztion of four distinct subsets of gamma/delta T cells. Two of the subsets show unique tissue preferences in the animal. For example, one subset preferentially accumulates in the spleen, whereas another one preferentially localizes in sites of inflammation. The molecular basis for the tissue tropism of these subsets is currently being studied.

Impacts
(N/A)

Publications

  • Abrahamsen, M.S., Lancto, C., Walcheck, B., and Jutila, M.A. 1997. Recruitment of leukocytes ot Cryptosporidium-infected tissues.
  • Jones, W., et al. 1997. Comparison of E-selectin binding glycoprotein ligands on human lymphocytes and neutrophils, and bovine gamma/delta T cells. J. Immunol. 159:3574-3583.
  • Jutila, M.A. and Kurk, S. 1996. Human lymphocytes roll on immobilized neutrophils: Role of PSGL-1 in leukocyte on leukocyte rolling.
  • Wilson, E., et al. 1997. Preferential tissue localization of bovine gamma/delta T cell subsets defined by anti-T-cell receptor for antigen antibodies. Eur. J. Immunol. submitted.
  • Hickey, M.J., et al. 1997. Tumor necrosis factor-alpha induces leukocyte recruitment of different mechanisms in vivo and in vitro. J. Immunol. 158:3391-3400.
  • Seekamp, A., et al. 1997. The effect of anti-L-selectin (EL-246) on remote lung injury after infrarenal ischemia/reperfusion. Shock.
  • McEvoy, L.M., et al. 1997. Anti-CD43 inhibits monocyte-endothelial cell adhesion in inflammation and atherogenesis. Blood. in press.
  • Jutila, M.A., Wilson, E., and Kurk, S. 1997. Identif. and charac. of an adhesion molecule that mediates leukocyte rolling on 24-hour cytokine stimulated bovine endothelial cells under flow conditions. J. Exp. Med., 186:1701-1711.


Progress 01/01/96 to 12/30/96

Outputs
This project provides funds for salary in support of our overall studies on bovine gamma/delta T cells. Our experimental aims relating specifically to the Hatch project deal with the analysis of a number of new gamma/delta T cell-specific monoclonal antibodies we have recently generated. We have defined antibodies that recognize WC1 and the gamma/delta T cell receptor (TCR) for antigen. Our TCR antibodies are quite interesting in that they define at least four distinct subsets within the overall gamma/delta T cell pool. We have also identified a new lineage-specific surface antigen (GD3.5 antigen) on gamma/delta T cells, distinct from previously described antigens. In the past year, new g/d T cell-specific MAbs have been generated which apparently react with carbohydrate determinants. These new antibodies are currently being characterized in a series of functional assays. These new antibodies are providing powerful tools in our NRI-USDA funded studies.

Impacts
(N/A)

Publications

  • Jutila, M.A. 1996. Gamma/delta T cell/endothelial cell interactions. Vet Immunol. Immunopath., 54:105.
  • Jutila, M.A. 1996. Analysis of bovine g/d T cell interactions with E-, P-, and L-selectin: Characterization of lymphocyte on lymphocyte rolling and the effectsof O-glycoprotease. J. Immunol. 156:289.
  • Jones, W., B. Walcheck, and M.A. Jutila. 1996. Generation of a new g/d T cell-specific monoclonal antibody (GD3.5): Biochemical comparisons of GD3.5 antigen with the previously described WC1 family. J. Immunol. 156:3772.
  • Jutila, M.A. 1996. Lymphocyte trafficking. ImmunoToxicology, invited review, in press.
  • Jones, W., Watts, G., Robinson, M., Vestweber, D., and Jutila, M.A. 1996. Comparison of E-selectin binding ligands on bovine g/d T cells and human lymphocytes and neutrophils. Submitted.


Progress 01/01/95 to 12/30/95

Outputs
This project provides funds for salary in support of our overall studies on bovine gamma/delta T cells. Our experimental aims related specifically to the Hatch project deal with the analysis of a number of new gamma/delta T cell specific monoclonal antibodies we have recently generated. We have defined antibodies that recognize WC1 and the gamma/delta T cell receptor (TCR) for antigen. Our TCR antibodies are quite interesting in that they define at least four distinct subsets within the overall gamma/delta T cell pool. We have also identified a new lineage-specific surface antigen (GD3.5) antigen) on gamma/delta T cells, distinct from previously described antigens, that may serve as ligand for P-selectin expressed by inflamed platelets. These new antibodies are providing powerful tools in our NRI-USDA funded studies.

Impacts
(N/A)

Publications


    Progress 01/01/94 to 12/30/94

    Outputs
    We began analysis of the vascular/platelet adhesion protein GMP-140, called P-selectin, in chronic inflammation of ruminants. We intended to follow up on the characterization of antibodies, previously made in the laboratory, that we originally believed to recognize goat, sheep, and cow P-selectin. In our original proposal, our preliminary regulation and biochemical data strongly supported that 2 new antibodies recognized ruminant P-selectin. This turned out to not be the case. Neither antibody recognized human P-selectin cDNA transfected mouse L1/2 cells (both antibodies crossreact with human), whereas a panel of anti-P-selectin antibodies provided by Eugene Butcher (Stanford University) did. We are still attempting to define the nature of the antigens these antibodies recognize. We then worked to identify anti-P-selectin antibodies that crossreact in ruminant. We found that WAPS provided by Eugene Butcher) stains bovine P-selectin. We then found that g/d T cells are the predominat ruminant lymphocyte that avidly binds P-selectin (see desc below). Determining if P-selectin can be expressed for prolonged periods on the surface of endothelial cells was our major objective. We confirmed the work of others and showed that P-selectin is expressed on certain venules in sites of chronic inflammation, included inflamed synovitis, as predicted in our original proposal. We have also attempted to define the in vitro conditions for chronic expression of P-selectin on cultured endothelial cells.

    Impacts
    (N/A)

    Publications

    • JUTILA, M.A., R.F. BARGATZE, S. KURK, R.A. WARNOCK, N. EHSANI, S. WATSON, AND B. WALCHECK. 1994. Cell surface P-and E-selectin support shear-dependent rolling of bovine g/d T cells. J. Immunol. 153:3917-3928.
    • WALCHECK, B., G. WATTS, AND M.A. JUTILA. 1993. Gamma/delta T cells bind E-selectin via a novel glycoprotein receptor: Characterization of the first lymphocyte-E-selectin interaction in an animal model. J. Exp. Med. 178:331-335.
    • JUTILA, M.A. 1994. Function and regulation of leukocyte homing receptors. J. Leuk. Biol. 55:133-140.
    • BARGATZE, R., S. KURK, G. WATTS, T. KISHIMOTO, C.A. SPEER, AND M.A. JUTILA. 1994. In vivo/in vitro funct examin of conserved epitopes of L-and E-selectin crucial for leukocyte-endothelial cell interactions. J Immunol 152:5814-5825.
    • BARGATZE, R.F., S. KURK, E.C. BUTHER, AND M.A. JUTILA. 1994. Neutrophils roll on adherent neutrophils via L-selectin on the rolling cells. J. Exp. Med. 180:1785-1792.
    • WALCHECK, B., AND M.A. JUTILA. 1994. Gamma/Delta T-cells express high levels of functional peripheral lymph node homing receptor (L-selectin). Intl. Immunol. 6:81-91.


    Progress 01/01/93 to 12/30/93

    Outputs
    The intent of the HATCH-funded project (419) is characterization of the role of GMP-140 in chronic inflammation. We have made considerable progress in these studies by characterizing key adhesion proteins in ruminants, developing new in vitro and in vivo functional assays, and generating new monoclonal antibodies. We have found that ruminant gamma/delta T cells avidly bind GMP-140 (now called P-selectin), and the vascular selectins control the migration of these T cells into sites of inflammation. We have done in vivo antibody blocking studies to show the blocking selectin-mediated adhesion inhibits the accumulation of gamma/delta T cells in inflamed skin of cattle. We have also defined the expression of vascular adhesion molecules in the inflamed synovium and confirmed earlier reports that P-selectin is expressed at high levels in this tissue. Publications related to this work are listed below.

    Impacts
    (N/A)

    Publications

    • WALCHECK, B. AND JUTILA, M.A. 1993. Regulation of leukocyte entry into inflameda tissues:...In "Structure and function of molecules involved in leukocyte adhesion", Faanes, R.B., et al., editors. Springer-Verlag, New York. in pres.
    • JUTILA, M.A. 1993. Selectins in leukocyte extravasation: function of a common epitope on L- and E-selectin. In "Advances in Pharmacology." Academic Press, Inc. San Diego, CA. in-press.
    • WALCHECK, B., AND JUTILA, M.A. 1993. Gamma/delta T-cells express high levels of functional peripheral lymph node homing receptor (L-selectin). International Immunol., in-press.
    • WALCHECK, B., AND JUTILA, M.A. 1993. Gamma/delta T cells bind E-selectin via a novel glycoprotein: first characteization of a lymphocyte-E-selectin interaction in an animal model. J. Exp. Med. 178:853-863.
    • JUTILA, M.A. 1993. The Selectin family. In "Biomembranes." Vol. 3. A.G. Lee, editor. JAI Press, Inc., Greenwich, Conn. in-press.
    • JUTILA, M.A. 1993. Generation of monoclonal antibodies against glycoproteins in host tissues. In "Molecular Biology of Pathogenic Fungi: A Laboratory Manual," in-press.
    • JUTILA, M.A. 1993. Function and regulation of selectins: A new family of leukocyte and endothelial cell adhesion proteins. In "Advances in Molecular and Cell Biology." D. Coleman, Editor, JAI Press, Inc., in press.


    Progress 01/01/92 to 12/30/92

    Outputs
    The project on the "Role of GMP-140 in chronic inflammation" began in October, 1992. Since that time, we have found that our SH43 and GSEC435 monoclonal antibodies recognize antigens that have all of the characteristics of GMP-140 but represent novel molecules. Neither antibody stained GMP-140 cDNA transfectants which represented one of our specific aims. In collaboration with Dr. Eugen Butchere's group at Stanford University, we have obtained 2 new monoclonal antibodies that react with "bona fida" GMP-140 in humans and ruminants. We have established novel in vitro functional assays to evaluate the effects of these new antibodies, as well as our SH43 and GSEC435 antibodies. We are also pursuing the characterization of the 435 antigen, that is expressed within the cytoplasm of endothelial cells in vitro and in vivo at levels far greater than GMP-140. The immunoaffinity purified antigen is not recognized by anti-GMP-140 or anti-vonWillibrand's factor antibodies. In preliminary analysis, the antibody appears to block monocyte adhesion to venules in the inflamed goat synovium.

    Impacts
    (N/A)

    Publications

    • NO PUBLICATIONS REPORTED THIS PERIOD.