Author
Urban, Joseph | |
Dawson, Harry | |
Solano-Aguilar, Gloria | |
Li, Robert | |
SHEA-DONOHUE, T - University Of Maryland |
Submitted to: Book Chapter
Publication Type: Book / Chapter Publication Acceptance Date: 1/5/2014 Publication Date: 4/1/2015 Citation: Urban Jr, J.F., Dawson, H.D., Solano Aguilar, G., Li, R.W., Shea-Donohue, T. 2015. Appendix III - Collection, handling and analysis of specimens for studies of mucosal immunity in animals of veterinary importance. In: Mucosal Immunity,4th edition. Mestecky, Strober, Russell, Cheroutre, Lambrecht, and Kelsall (eds). Elsevier. p. 2369–2391. DOI:10.1016/B978-0-12-415847-4.15003-7. Interpretive Summary: The validity of data in science depends on the reliability of the collection procedure and the methods used for data analysis. This becomes a major challenge in studies of mucosal immunology since samples for analysis come from various anatomical sources which differ remarkably in their content and properties. This appended chapter describes procedures for the collection and analysis of body fluids and secretions from especially large animals of veterinary importance. Because of the size and volume of secretions that can be obtained, the cellular fraction can be recovered in a quantity that allows for more careful analysis by flow cytometry and for functional studies of subpopulations of leucocytes. This appendix also details surgical procedures and interventions not often used in laboratory animals that allow periodic sampling without sacrificing the animal. Because of the diversity of large animals of veterinary importance, the procedures described are based on using one of several common species. However they may be applicable to many species. Some procedures such as measures to inhibit proteases, storage procedures, compensating for blood contamination or transudation and expressing antibody responses in body fluids, are issues faced by all who work in the field of mucosal immunology and should have universal value. While we do not describe methods like FCM or RT-PCR, we do provide references to databases for reagents that are available for studies in large animals of veterinary importance. The use of large transgenic animals for research and a source of humanized antibodies is not reviewed. Rather, readers are directed to other sources. Technical Abstract: The validity of data in science depends on the reliability of the collection procedure and the methods used for data analysis. This becomes a major challenge in studies of mucosal immunology since samples for analysis come from various anatomical sources which differ remarkably in their content and properties. This appended chapter describes procedures for the collection and analysis of body fluids and secretions from especially large animals of veterinary importance. Because of the size and volume of secretions that can be obtained, the cellular fraction can be recovered in a quantity that allows for more careful analysis by flow cytometry and for functional studies of subpopulations of leucocytes. This appendix also details surgical procedures and interventions not often used in laboratory animals that allow periodic sampling without sacrificing the animal. Because of the diversity of large animals of veterinary importance, the procedures described are based on using one of several common species. However they may be applicable to many species. Some procedures such as measures to inhibit proteases, storage procedures, compensating for blood contamination or transudation and expressing antibody responses in body fluids, are issues faced by all who work in the field of mucosal immunology and should have universal value. While we do not describe methods like FCM or RT-PCR, we do provide references to databases for reagents that are available for studies in large animals of veterinary importance. The use of large transgenic animals for research and a source of humanized antibodies is not reviewed. Rather, readers are directed to other sources. |