Progress 06/01/05 to 05/31/10
Outputs
Progress Report Objectives (from AD-416) 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. 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. 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 (from AD-416) 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. A National Pork Board (NPB) grant has funded this Specific Cooperative Agreement (SCA) with the Flanders Interuniv. for Biotech and the Free Univ. in Brussels, Belgium. This SCA has helped to expand the immune reagent toolkit for pigs; it utilizes the Belgian expertise to express segments of cloned swine antibody, or immunoglobulin (Ig), genes as recombinant camelid-swine IgGs. Swine produce IgG antibodies in response to infection or vaccination, but exactly which IgG subclass (IgG1 - IgG6) is involved in each function remains unknown. To test the functional properties of swine IgG subclasses, it is necessary to have purified IgG subclass proteins and monoclonal antibodies (mAb) that can specifically recognize each separate subclass protein. Since porcine IgG subclass proteins cannot be separated by biochemical methods, our Belgian colleagues have expressed portions of most of 6 of the 11 known swine IgG subclass genes in vitro using a camel Ig chimeric protein expression system. A new set of these proteins is being prepared using updated protocols developed in Belgium; once available these will be shipped to the U.S. to pursue the NPB grant third objective, i.e., to develop new mAbs that are specific for each of the expressed IgG proteins. Overall, the National Pork Board Grant (1265-32000-088-01R) and its associated SCAs (1265-32000-088-02S and -03S) will help develop, for all swine researchers, mAb reactive with the different IgG subclass proteins. These reagents will give researchers better ways to define, and ultimately target, the level of specific antibody response to infectious disease and vaccines. Discussing progress of this grant was in regular email and phone contact with the participating scientists at the Univ. Iowa and at the Flanders Interuniv. for Biotech and Free Univ., Brussels, Belgium.
Impacts
(N/A)
Publications
|
Progress 10/01/08 to 09/30/09
Outputs
Progress Report Objectives (from AD-416) 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. 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. 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 (from AD-416) 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. Significant Activities that Support Special Target Populations This SCA has helped to expand the immune reagent toolkit for pigs; it utilizes the Belgian expertise to express segments of cloned swine antibody, or immunoglobulin (Ig), genes as recombinant camelid-swine IgGs. Swine produce IgG antibodies in response to infection or vaccination, but exactly which IgG subclass (IgG1 - IgG6) is involved in each function remains unknown. To test the functional properties of swine IgG subclasses, it is necessary to have purified IgG subclass proteins and monoclonal antibodies (mAb) that can specifically recognize each separate subclass protein. Since porcine IgG subclass proteins cannot be separated by biochemical methods, our Belgian colleagues have expressed portions of most of these 6 swine IgG subclass genes in vitro using a camel Ig chimeric protein expression system. A new set of these proteins are being prepared using updated protocols developed in Belgium; once available these will be shipped to the U.S. to develop new mAbs that are specific for each of the expressed IgG proteins. Overall, the National Pork Board Grant (1265-32000-087-01R) and its associated SCAs (1265-32000-087-02S and -03S) will help develop mAbs reactive with the different IgG subclass proteins. These reagents will give researchers better ways to define, and ultimately target, the level of specific antibody response to infectious disease and vaccines. Progress of this grant was monitored by regular email and phone contact with the participating scientists at the Univ. Iowa and at the Flanders Interuniv. for Biotech and Free Univ., Brussels, Belgium.
Impacts
(N/A)
Publications
|
Progress 10/01/06 to 09/30/07
Outputs
Progress Report Objectives (from AD-416) 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. 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. 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 (from AD-416) Dr. Serge Muyldermans in Belgium will express each swine IgG subclass protein using his camel Ig expression system. Dr. John Butler at Univ. 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 he will characterize the reactivity of known anti-swine Ig mAb reagents with each Ig gene product. Significant Activities that Support Special Target Populations This report documents research conducted under a Specific Cooperative Agreement between ARS and the Free Univ., Brussels, Belgium, funded through National Pork Board (NPB) Grants to USDA ARS BARC APDL and University of Iowa scientists. Additional details of research can be found in the report for the NPB grant, 1265-32000-079-010R, and the parent project, 1265-32000-079-00D, "Strategies to Control Swine Parasites Affecting Food Safety." National Pork Board (NPB) grant has funded this Specific Cooperative Agreement (SCA) with the Free Univ. in Brussels, Belgium. This SCA has helped to expand the immune reagent toolkit for pigs; it utilizes the Belgian expertise to express segments of cloned swine antibodies or immunoglobulins (Igs), as camelid-swine IgGs. Swine produce Ig antibodies in response to infection or vaccination but exactly which IgG subclasses (IgG1 - IgG6) are involved in each function remains unknown for the lack of reagents. As outlined in the associated annual report #1265-32000-079- 11S our colleagues at the Univ. Iowa have already characterized and cloned eight different swine IgG genes, and sent those genes and their sequences to Belgium. Our Belgian colleagues have expressed most of these swine IgG subclass proteins in vitro using a camel Ig expression system. These single chain camelid-swine IgGs each have the same Ig variable region, which binds lysozyme, attached to different swine IgG constant regions. These proteins have been shipped to the U.S. and area now being used for our second objective, to characterize the reactivity of all currently available monoclonal antibodies (mAb) anti-swine Ig, and our third objective to begin to develop new mAbs that are specific for each of the expressed IgG proteins. The US labs are jointly characterizing the reactivity of each anti-swine IgG mAb reagent with each camelid IgG gene product. Overall, this National Pork Board Grant (1265-32000-064-10R) and its associated SCAs will help develop, for all swine researchers, mAb reactive with the different IgG subclass proteins. These reagents will give researchers better ways to define, and ultimately target, the level of specific antibody (Ig) responses to infectious disease and for vaccine studies. National Program 103 Component 2: Genetic and biological determinants of disease susceptibility Discussions on progress and plans for this SCA have been in regular email contacts with the participating Belgian lab as they developed reagents.
Impacts
(N/A)
Publications
|
Progress 10/01/05 to 09/30/06
Outputs
Progress Report 4d Progress report. This report serves to document research conducted under a Specific Cooperative Agreement between ARS and the Free Univ., Brussels, Belgium, funded through National Pork Board (NPB) Grants to USDA ARS BARC APDL and University of Iowa scientists. Additional details of research can be found in the report from the NPB grants, 1265-32000-079-010R, and the parent project, 1265-32000-079-00D: Strategies to Control Swine Parasites Affecting Food Safety. This National Pork Board (NPB) grant funded Specific Cooperative Agreement (SCA) with the Free Univ. in Brussels, Belgium, will help expand the immune toolkit for pigs, by utilizing their expertise to express cloned swine antibodies or immunoglobulins (Igs) as camelid-swine IgGs. Swine produce Ig antibodies in response to infection or vaccination but exactly which IgG subclasses (IgG1 - IgG6) are involved in each function remains unknown for the lack of reagents. As outlined in the associated
annual report #1265-32000-079-11S our colleagues at the Univ. Iowa have already characterized and cloned the different swine IgG genes, and sent those genes and their sequences to Belgium. Our Belgian colleagues will express each swine IgG subclass proteins in vitro using a camel Ig expression system. These single chain camelid-swine IgGs will have the same variable region and different IgG constant regions. These will then be used for our second objective, to characterize the reactivity of all currently available monoclonal antibodies (mAb) anti- swine Ig, and our third objective to begin to develop new mAbs that are specific for each of the expressed IgG proteins. Our labs will jointly characterize the reactivity of each anti-swine IgG mAb reagents with each IgG gene product. Overall, this National Pork Board Grant (1265-32000-064- 10R) and its associated SCAs will help develop, for all swine researchers, mAb reactive with the different IgG subclass proteins. These reagents will
give researchers better ways to define, and ultimately target, the level of specific antibody (Ig) responses to infectious disease and for vaccine studies.
Impacts
(N/A)
Publications
|
|