Progress 09/01/03 to 08/31/07
Outputs
OUTPUTS: This project was focused on the analysis of myeloid cell associated genes and their products in bovine gamma/delta T cells. We have performed a comprehensive analysis of over 20 myeloid cell genes in sorted bovine gamma/delta T cells. Transcripts were found for all TLRs, scavenger receptors, CD11b, and a number of other genes. We systematically examined the function of each of these gene products. We first showed that bovine gamma/delta T cells express functional CD36, which is regulated by phorbol ester activation. Gamma/delta T cell CD36 facilitates TLR2 recognition of LTA. We have also confirmed that gamma/delta T cells express NRAMP, NOD2 and the MARCO scavenger receptor, which are the focus of our current analyses. Analysis of the BLIMP-1 transcription factor, which is important in the differentiation of myeloid cells and B cells, is also being pursued in this project. In addition to the study of the regulation of Blimp-1 in bovine T cells, we have recently confirmed
expression in human gamma/delta T cells. Using well defined molecular approaches available in the human system, we have now confirmed functional activity of gamma/delta T cell Blimp-1 in gel shift assays. Specifically, gamma/delta T cell Blimp-1 binds the human cIITA and cMYC Blimp-1 binding sites. The interaction is highly conserved, since bovine Blimp-1 also binds the same human promoters. Though not included in the original proposal, in the past year we have explored the use of nanotechnology to delivery antigens/agonists to bovine cells. Our current experiments are focused on delivering PAMPs via viral nanoparticles, as an approach to selectively stimulated bovine gd T cells. We have also recently tested a novel inhibitor of T cell signaling. A grant on NRAMP-1 in gamma/delta T cells was submitted in 2007.
PARTICIPANTS: Mark Jutila served as PI on this project. Jill Graff completed her MS dissertation work on this project and then moved into a Research Associate position. She led most of the work done on this project. Kirk Lubick worked as a MAES funded research associate on this project. Edward Schmidt was a collaborator who worked closely with Ms Graff on the transcription factor studies on this project. Jodi Hedges worked a short time on this project as a postdoc. A number of undergraduate students spent short periods of time on this project.
Impacts
This series of experiments has shed light on common gene expression patterns in bovine myeloid cells and gamma/delta T cells. Identifying and characterizing funtional myeloid cell gene products in gamma/delta T cells may lead to the development of novel approaches to control the activity of these cells.
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. Targeting of cancer cells with ferrimagnetic ferritin cage nanoparticles. J. Amer. Chem. Soc.,128:16626.
- Kaiser, C.R., Flenniken, M., Gillitzer, E., Harmsen, A.L., Harmsen, A. G., Jutila, M.A., Douglas, T.,and Young, M.J. 2007. Biodistribution studies of protein cage nanoparticles demonstrate broad tissue distribution and rapid clearance in vivo. Intern. J. Nanomedicine, 2: 715.
- 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. Animal Health Research Reports, in press.
- Ren, Y., Strobel, G., Graff, J., Jutila, M., Park, S.G., Gosh, S., Teplow, D., Condron, M., Pang, E., and Hess, W.M. 2007. Volutellin A, an immunosuppressive peptide from Volutella sps,. Microbiology, in press.
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Progress 01/01/06 to 12/31/06
Outputs
This project is focused on the analysis of myeloid cell associated genes and their products in bovine gamma/delta T cells. We have performed a comprehensive analysis of over 20 myeloid cell genes in sorted bovine gamma/delta T cells. Transcripts were found for all TLRs, scavenger receptors, CD11b, and a number of other genes. We have started to systematically examine the function of each of these gene products. We first showed that bovine gamma/delta T cells express functional CD36, which is regulated by phorbol ester activation. We have now found that certain TLR agonists will also regulate CD36 on gamma/delta T cells. Gamma/delta T cell CD36 facilitates TLR2 recognition of LTA. We have also confirmed that gamma/delta T cells express NRAMP, NOD2 and the MARCO scavenger receptor,which are the focus of our current analyses. In the past year, we showed that bovine gamma/delta T cells respond to the NOD2 agonist, muramyl dipeptide. The NOD2 response leads to priming of
the gamma/delta T cells, which greatly increases downstream proliferative responses to IL-2 and IL-15. We recently have started an analysis of CD11b on gamma/delta T cells in which we have shown its regulation in response to plant polyphenols is opposite to the regulation of CD11b on monocytes. Our detailed functional analysis of BLIMP-1 suggest that it may function in a unique fashion in gamma/delta T cells, in that we have now confirmed its expression is upregulated at a period o time in which gamma/delta T cells are actively proliferating in response to IL-2. This was not expected, since the literature shows BLIMP-1 to be a repressor of proliferation.
Impacts
This series of experiments will shed light on common gene expression patterns in bovine myeloid cells and gamma/delta T cells. Indentifying and characterizing funtional myeloid cell gene products in gamma/delta T cells may lead to the development of novel approaches to control the activity of these cells
Publications
- 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
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Progress 01/01/05 to 12/31/05
Outputs
This project is focused on the analysis of myeloid cell associated genes and their products in bovine gamma/delta T cells. We have performed a comprehensive analysis of over 20 myeloid cell genes in sorted bovine gamma/delta T cells. Transcripts were found for all TLRs, scavenger receptors, CD11b, and a number of other genes. We have started to systematically examine the function of each of these gene products. We first showed that bovine gamma/delta T cells express functional CD36, which is regulated by phorbol ester activation. Gamma/delta T cell CD36 facilitates TLR2 recognition of LTA. We have also confirmed that gamma/delta T cells express NRAMP, NOD2 and the MARCO scavenger receptor,which are the focus of our current analyses. Analysis of the BLIMP-1 transcription factor, which is important in the differentiation of myeloid cells and B cells, is also being pursued in this project. In addition to the study of the regulation of Blimp-1 in bovine T cells, we have
recently confirmed expression in human gamma/delta T cells. Using well defined molecular approaches available in the human system, we have now confirmed functional activity of gamma/delta T cell Blimp-1 in gel shift assays. Specifically, gamma/delta T cell Blimp-1 binds the human cIITA and cMYC Blimp-1 binding sites.
Impacts
This series of experiments will shed light on common gene expression patterns in bovine myeloid cells and gamma/delta T cells. Indentifying and characterizing funtional myeloid cell gene products in gamma/delta T cells may lead to the development of novel approaches to control the activity of these cells.
Publications
- Lubick, K and Jutila, M.A.. 2005. LTA Recognition by Bovine gamma/delta T cells Involves CD36. Journal of Leukocyte Biology, in press.
- 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, and MA Jutila. 2005. Response of gammadelta T cells to the NOD2 agonist muramyl dipeptide. FASEB J. (Abstract) 826.15.
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Progress 01/01/04 to 12/31/04
Outputs
This project is focused on the analysis of myeloid cell associated genes and their products in bovine gamma/delta T cells. We have performed a comprehensive analysis of over 20 myeloid cell genes in sorted bovine gamma/delta T cells. Transcripts were found for all TLRs, scavenger receptors, CD11b, and a number of other genes. We have started to systematically examine the function of each of these gene products. We have now shown that bovine gamma/delta T cells express functional CD36, which is regulated by phorbol ester activation. Gamma/delta T cell CD36 facilitates TLR2 recognition of LTA, but not peptidoglycan. We have also confirmed that gamma/delta T cells express NRAMP and the MARCO scavenger receptor. Analysis of the BLIMP-1 transcription factor, which is important in the differentiation of myeloid and B cells, is also being pursued in this project. We have shown that BLIMP-1 is regulated in gamma/delta T cells and have established RNA interference techniques
to downregulate BLIMP-1 transcripts in primary gamma/delta T cells as an approach to analyze its function. New antibody reagents have been generated to study BLIMP-1 protein in bovine cells.
Impacts
This novel series of experiments will shed light on common gene expression patterns in bovine myeloid cells and gamma/delta T cells. Analyses of these gene products may shed light into novel approaches to altering the function of gamma/delta, which may lead to the development of new adjuvants to enhance both innate and adpative immunity.
Publications
- 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., 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.
- Lubick, K. and Jutila, M.A. 2004. Expression of CD36 on bovine gamma/delta T cells. FASEB Abstract, submitted
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Progress 01/01/03 to 12/31/03
Outputs
This is a new project that follows on preliminary data generated in our functional genomic analysis of bovine gamma/delta T cells. In those studies, we found a link between gamma/delta T cells and myeloid cells, based on common gene expression patterns. The intent of this new project is to determine whether any of the "myeloid" cell gene products actually are functionally important in gamma/delta T cells. One particularly intriguing gene is BLIMP-1 which is a transcription factor that regulates myeloid cell development. BLIMP-1 mRNA is expressed in bovine gamma/delta T cells. Experiments will be done to determine if functional protein is also expressed. We have succeeded in generating a polyclonal antisera that reacts with bovine BLIMP-1 and preliminary experiments confirm BLIMP-1 protein expression in bovine gamma/delta T cells
Impacts
This novel series of experiments will shed light on common gene expression patterns in bovine myeloid cells and gamma/delta T cells. Analyses of these gene products may shed light into novel approaches to altering the function of gamma/delta, which may lead to the development of new adjuvants to enhance both innate and adpative immunity.
Publications
- No publications reported this period
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