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Swine disease and vaccine research has been advanced by the development of sophisticated tools to measure physiologic parameters associated with immunity, pathology, and disease prevention. Our goal for this National Pork Board (NPB) funded grant (#05-015) is to expand the immune toolkit for pigs, by developing and characterizing reagents that can be used to identify and quantify a major class of immune proteins, the immunoglobulins (Igs). This SCA will cover the work in Dr. John Butler's laboratory at the Univ. Iowa. He will be producing new monoclonal antibody (mAb) reagents against swine Igs. Swine produce antibodies in response to infection or vaccination but exactly which Ig classes, in particular IgG subclasses (IgG1 - IgG6) are involved in each function remains unknown for the lack of reagents. Reseachers require Ig subclass specific reagents to determine Ig function; diagnostic laboratories use them to measure Ig levels and specific antibody responses according to isotype. Currently most laboratories and investigators rely on polyclonal antisera that are tedious to prepare, lack immortality and vary between batches. We propose to develop monoclonal antibody (mAb) reagents that uniquely recognize the various Ig isotypes and IgG subisotypes, or subclasses. <P>To accomplish this, our Belgian colleagues will first express all swine IgG subclass and minor isotype proteins in vitro using a camel Ig expression system. This is necessary before we can perform our second objective, to characterize the reactivity of all currently available mAb anti-swine Ig. The expressed Ig proteins can also be used for our third objective, to produce new mAb at the Univ. Iowa facilities.<P> Our overall goal is to have a full panel of well-characterized mAb that react specifically with each swine Ig isotype and IgG subclass. These mAb reagents will refine swine disease diagnostic tests and enable scientists to more accurately compare results among laboratories. We expect that use of these mAb anti-swine Ig reagents will prove valuable in studies to reveal the functions of each swine Ig isotype and subclass, thus opening up new understanding of disease control mechanisms and vaccine responses.
Approach: At the Free University in Brussels, Belgium they will express each swine IgG subclass protein using their camel Ig expression system. The Univ. of Iowa will use these Ig proteins to begin to develop new mAbs that are specific for each of the expressed Ig proteins. In collaboration with our lab at BARC they will characterize the reactivity of known anti-swine Ig mAb reagents with each Ig gene product.