2012 Annual Report
1a.Objectives (from AD-416):
U.S. Veterinary Immune Reagent Network (US VIRN) www.vetimm.org was established in 2005 and represents a broad community plan to systematically address the immunological reagent gap for the U.S. veterinary immunology research community. It includes the following groups: ruminants (concentrating on cattle), swine, poultry (primarily chickens with some evaluation of reagents for turkeys), horses, and aquaculture species (concentrating on channel catfish and trout). For this renewal the groups will develop sets of reagents, i.e. monoclonal antibodies (mAb), that can identify the major leukocyte cell populations (T and B lymphocytes, macrophages, dendritic cells, neutrophils) to evaluate changes during disease and to allow scientists to manipulate them in vitro and in vivo. Such manipulations enable evaluation of the cell population’s role in protective immunity and immunopathology. In addition they will clone and expreess recombinant cytokines and chemokines (the soluble messenger molecules of the immune system) using mammalian, yeast, and bacterial systems. They will develop mAb to these proteins and their receptors to facilitate understanding of their contributions to disease and pathology. Finally, reagents to identify immunoglobulin (Ig) isotypes are needed since isotypes differ from one another functionally and thus in their effect on disease outcome. The reagents will benefit a large group of U.S. and international researchers including veterinary immunologists, pathologists and microbiologists.
1b.Approach (from AD-416):
Prioritization of reagents (tools) to be produced will be established by surveying experts within the relevant scientific community and commercial companies as well as cooperating with similar international efforts to avoid redundancy and target the most useful reagents for research scientists. These reagents will include.
1)Cytokines: bioactive recombinant proteins as well as antibodies to them;.
2)mAbs to: Cytokine receptors, Ig isotypes, T cell receptor (TCR) constant and variable regions, Toll like (TLR) receptors, and other pattern recognition receptors and regulatory receptors including NKG2 family members and KIR. The mAb will have the following properties:.
1)For anti-cytokines mAb: mAb pairs for ELISA, ELISpot and fluorescent microsphere immunoassay (FMIA), or Luminex, assays. For these assays mAb need to react with 2 different epitopes on the same molecule. Additionally single mAb are needed for intracellular staining for cytokines;.
2)For anti-receptor mAb: mAb that block function and signaling molecules (e.g. TLR); and.
3)For cell surface or CD molecule mAbs: mAb that react with native molecules to stain viable cells for flow cytometric analyses; as well as mAb that react in fixed tissue sections. The reagents will be provided to the scientific community by depositing in commercial cell banks and encouraging generation and distribution through commercial vendors.
Under this Agreement funded through a USDA/NIFA/AFRI grant to the University of Massachusetts-Amherst (UMA), the U.S. Veterinary Immunological Reagents Network (VIRN) was established to address the dearth of immunological reagents for food animal species. ARS researchers at Beltsville, MD, (BARC), worked to develop immune reagents for swine to assist researchers to probe normal immune responses and responses to infectious diseases, and to improve vaccine design. BARC researchers, working with our commercial partner, Kingfisher Biotech Inc., have expressed several new recombinant swine immune proteins in yeast. These included recombinant cytokines, e.g. the regulatory cytokines, interleukin-17A (IL-17A) and IL-17F. Bioassays performed at BARC affirmed that they were bioactive. Efforts are underway with our collaborators at UMA, to use these proteins for immunizations and monoclonal antibody development. Panels of the resultant monoclonal antibodies have been screened for specific anti-cytokine reactivity. Products generated by the VIRN will be used by researchers, veterinarians, vaccine manufacturers, and other commercial sources to improve animal health and develop better vaccines.