The development and validation of methods for evaluating the immune system in preweaning piglets.
The preweaning piglet has been found to be a valuable research model for testing ingredients used in infant formula. As part of the safety assessment, the neonates' immune system is an important component that has to be evaluated. In this study three concurrent strategies were developed to assess immune system status. The methods included (1) immunophenotying to assess circulating innate immune cell populations, (2) monitoring of circulating cytokines, particularly in response to a positive control agent, and (3) monitoring of localized gastrointestinal tissue cytokines using immunohistochemistry (IHC), particularly in response to a positive control agent. All assays were validated using white papers and regulatory guidance within a GLP environment. To validate the assays precision, accuracy and sample stability were evaluated as needed using a fit for purpose approach. In addition animals were treated with proinflammtory substances to detect a positive versus negative signal. In conclusion, these three methods were confirmed to be robust assays to evaluate the immune system and GIT-specific immune responses of preweaning piglets.
MPI Research, Mattawan, MI, 49009, USA. Electronic address: Brandon.Zeigler@mpiresearch.com.
Brandon M
BM
Zeigler
Beckman Coulter, Southfield, MI, 48033, USA.
Mark
M
Cameron
MPI Research, Mattawan, MI, 49009, USA.
Keith
K
Nelson
MPI Research, Mattawan, MI, 49009, USA.
Kristi
K
Bailey
TOXpertise, Princeton, NJ, 08540, USA.
Myra L
ML
Weiner
Abbott Laboratories, Columbus, OH, 43219, USA.
Brinda
B
Mahadevan
Experimur, Chicago, IL, 60609, USA.
Bjorn
B
Thorsrud
D000818:Animals; D000831:Animals, Newborn; D015415:Biomarkers / Q000097:blood / Q000378:metabolism; D003433:Crosses, Genetic; D016207:Cytokines / Q000097:blood; D005260:Female; D005434:Flow Cytometry / Q000662:veterinary; D041981:Gastrointestinal Tract / Q000166:cytology / Q000254:growth & development / Q000276:immunology / Q000378:metabolism; D007113:Immunity, Innate; D018928:Immunity, Mucosal; D007150:Immunohistochemistry / Q000662:veterinary; D016130:Immunophenotyping / Q000662:veterinary; D008297:Male; D008824:Michigan; D018448:Models, Immunological; D009092:Mucous Membrane / Q000166:cytology / Q000254:growth & development / Q000276:immunology / Q000378:metabolism; D055550:Protein Stability; D015203:Reproducibility of Results; D034421:Sus scrofa / Q000097:blood / Q000254:growth & development / Q000276:immunology* / Q000378:metabolism; D018675:Toxicity Tests
D015415:Biomarkers; D016207:Cytokines
ELISA; Flow cytometry; Immunohistochemistry (ICH); Immunophenotyping (IP); Immunotoxicology; Porcine