U.S. Veterinary Immune Reagent Network

Swine

Gene Sequences

Comparative Alignments

Primers

Bioassays

References for Genes

Publications & Posters

Plans & Progress

Commercially Available Reagents

Recombinant proteins have been made for the following genes:

IL-4, IL-6, IL-8, IL-10, IL-13, IL-15, IL-17A, IL-17F, IL-22, IFNβ, IFNγ, CCL2, CCL3L1, CCL4, CCL5, CXCL9, CXCL10, CXCL11, TNF-α

Recombinant proteins are available from Kingfisher Biotech, Inc.

Recombinant proteins will be made for the following genes:

IL-7, IL-9, IL-21, IL-23

Please see 'Progress' button for detailed information about expression and bioactivity for these molecules.

Monoclonal antibodies have/will be made to:

Cytokines and Chemokines:

CCL2: protein production and purification completed in 2008 (Kingfisher); bioassay activity affirmed 2009 (BARC); hybridoma fusion and mAb screening completed 2009 (Cornell); Luminex assay developed 2009 (Cornell, BARC); manuscript preparation 2012 (Cornell, BARC).

CCL3L1: protein production and purification completed in 2009 (Kingfisher); bioassay activity affirmed 2009 (BARC); combined chemokine manuscript preparation 2013 (BARC).

CCL4: protein production and purification completed in 2009 (Kingfisher); bioassay activity affirmed 2009 (BARC); combined chemokine manuscript preparation 2013 (BARC).

CCL5: protein production and purification completed in 2009 (Kingfisher); bioassay activity affirmed 2009 (BARC); combined chemokine manuscriot preparation 2013 (BARC).

CCL20: protein production and purification completed 2011 (Kingfisher); bioassay activity affirmed 2011 (BARC).

CXCL10: protein production and purification completed 2008 (Kingfisher); bioassay activity affirmed 2008 (BARC); combined species manuscript (Hudgens et al., Vet Immunol. Immunopathol. 141: 317-21, 2012); first hybridoma fusion 2008 but only 1 mAb identified (U Mass Amherst).

CXCL11: protein production and purification completed 2008 (Kingifsher); bioassay activity affirmed 2008; manuscript (Boyd et al., Vet Immunol. Immunopathol. 136: 170-5, 2010); hybridoma fusion completed 2008 (U Mass Amherst); only 1 mAb identified.

CXCL9: protein production and purification completed 2010 (Kingfisher); bioassay activity tested 2009 (BARC).

IFNα: protein production and purification completed 2010 (Kingfisher); hybridoma fusion completed 2012 (U Mass Amherst); as part of a DHS funded effort expanded effort to identify mAb reactive with specific IFNA1, IFNA6, IFNA9 gene products (Sang, Kansas State; Wagner, Cornell); several fusions cmpleted (Cornell) but no gene specific mAb yet identified (BARC).

IFNβ: protein production and purification completed 2008 (Kingfisher); positive bioassay activity (U Mass Amherst); further testing 2009/2010 (BARC); hybridoma fusion and mAb screening 2009 (U Mass Amherst, BARC); as part of DHS funded effort also screening IFNβ (Sang, Kansas State); fusion completed (Cornell); other option: test new cloning and screening of known anti-IFNβ hybridomas (U CT Garmendia; OIADC Grubman, BARC); low Ig production when grown in serum free medium (SFM); screenings continuing in 2013 (BARC).

IFNγ: protein production and purification completed 2010 (Kingfisher); positive bioassay activity 2011 (BARC); loss of crucial commercian mAb stimulated plans for production of new mAb in 2013 (Cornell, BARC).

IL-6: protein production and purification completed 2010 (Kingfisher); positive bioassay activity affirmed 2011 (BARC); hybridoma fusion completed 2012 with positive clones (U Mass Amherst); further characterization ongoing 2013 (BARC).

IL-13: protein production and purification completed 2008 (Kingfisher); bioassay activity testing (BARC) and retesting (Golde, PIADC) showed no/limited bioactivity; re-expression and purification 2010 (Kingfisher); hybridoma fusion completed 2011/2012 (U Mass Amherst); further characterization ongoing (BARC).

IL-15: protein production completed 2011 (Kingfisher); bioassay activity affirmed 2011 (BARC); commercial mAb available.

IL-17A: protein production and purification completed 2010 (Kingfisher); bioassay activity affirmed 2011 (BARC); hybridoma fusion completed 2012 with positive clones (U Mass Amherst); further characterization ongoing 2013 (BARC).

IL-17F: protein production and purification completed 2010 (Kingfisher); bioassay activity affirmed 2011 (BARC); hybridoma fusion cmpleted with positive clones 2012; 2nd fusion 2013 (U Mass Amherst); further characterization ongoing 2013 (BARC).

TNFα: protein production completed 2009 (Kingfisher); weak bioassay activity affirmed 2009 (BARC); commercial mAb available.

Cell Surface Molecules:

IL-4Rα (CD124): equine IL-4-swine IL-4R recombinant protein expressed 2008 (Cornell); hybridoma fusion completed 2008 (Cornell); mAb tested; best clones selected 2009 (BARC); specific bioactivity of selected mAb tested 2010/11/12 (Dawson, Lunney BARC); manuscript in revision 2013 (Cornell, BARC)

IL-13Rα (CD213A1): equine IL-4-swine IL-13R recombinant protein expressed 2009 (Cornell); mAb produced 2009 (cornell); original mAb tested; best clone selected 2009 (BARC).

TCRβ: First hybridoma fusion completed 2008 (Cornell); new equine IL-4-swine TCRβ recombinant protein production completed 2009; new hybridoma fusion performed 2010/11 (Cornell); supernatents screened (BARC); lack of success for this approach for mammalian TCRs resulted in termination of this effort in 2012.

TCRα: First hybridoma fusion completed 2008 (Cornell); new equine IL-4-swine TCRα recombinant protein production completed 2009; new hybridoma fusion performed 2010/11 (Cornell); supernatents screened (BARC); lack of success for this approach for mammalian TCRs resulted in termination of this effort in 2012.

CD45RO: Commercial peptide used for fusion 2008 (U Mass Amherst); mAbs tested (BARC); no strong positives; fusion repeated but no positive clones 2010; effort stopped.

CD19: Recombinant proteins expressed in mammalian cells 2012 (Cornell); CD19 transfectant proteins purified 2012 (Cornell); first fusion mAbs screened using CD19 transfectants 2012 (Cornell); panel of mAb screened on pig cells 2013 (BARC); further characterization ongoing 2013.

IFNAR1 IFNAR2: Recombinant proteins expressed in mammalian cells 2012 (Cornell); IFNA1 transfectants expressing high amounts of proteins; proteins purified and immunizations are ongoing 2013 (Cornell); mAb will be screened using IFNAR1 and IFNAR2 single transfectants and also co-transfectants of both receptor chains 2013 (Cornell); on stimulated pig cells (BARC).

NKp44: With added DHS funds have addressed mAb against natural killer (NK cells, specifically for natural cytotoxcity receptor (NCR2) or NKp44 or CD336 antigens; Recombinant proteins expressed in mammalian cells 2012 (Cornell); NKp44 transfectant proteins purified and immunizations are ongoing 2012 (Cornell); first fusion mAbs screened using NKp44 NCR1, NKp46, CD335 already available for pigs (Saalmueller, Austria).

NKp30: With added DHS funds have addressed mAb against natural killer (NK cells), specifically for NCR3 NKp30 or CD337 antigens; Recombinant proteins expression in mammalian cells planned 2013 (Cornell); further characterization ongoing 2013.

Immunoglobulin Isotype Specific mAbs:

With added DHS funds Butler, U Iowa, and Golde, PIADC, have addressed development of Ig isotype specific mAb. (For approach see: Butler et al. Molec. Immunol. 53:140-148, 2013). A commercial partner is performing the immunizations and fusions. Resultant mAb are being screened on camelid expressed porcine IgG proteins.

IgG3: Hybridomas screened with potential mAb identified; similar result for anti-pan PoIgG.

IgG5, IgG1, and IgG2-G4-G6 complex: promising hybridomas currently being subcloned for further screening.

IgG2: Immunized mouse with current fusion planned.

Click here to download the protocol for isolation of swine PBMC.

The Porcine Immunology and Nutrition Database can be found here: http://www.ars.usda.gov/Services/Services.htm?docid=6065.

Click here to download the survey form.
Then fill it out, and send it back to the appropriate species coordinator.

Last updated: March 2013