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This version was published on September
1, 2006
Journal of Biomolecular Screening, Vol. 11, No. 6,
664-671 (2006)
DOI: 10.1177/1087057106289876
Use of Early Passage Fetal Intestinal Epithelial Cells in Semi-High-Throughput Screening Assays: An Approach to Identify New Innate Immune System Adjuvants
Diana Buckner
Veterinary Molecular Biology, Montana State University, Bozeman
Suzanne Wilson
Veterinary Molecular Biology, Montana State University, Bozeman
Sandra Kurk
Veterinary Molecular Biology, Montana State University, Bozeman
Michele Hardy
Veterinary Molecular Biology, Montana State University, Bozeman
Nicole Miessner
Veterinary Molecular Biology, Montana State University, Bozeman
Mark A. Jutila
Veterinary Molecular Biology, Montana State University, Bozeman
Innate immune system stimulants (innate adjuvants) offer complementary approaches to vaccines and antimicrobial compounds to increase host resistance to infection. The authors established fetal bovine intestinal epithelial cell (BIEC) cultures to screen natural product and synthetic compound libraries for novel mucosal adjuvants. They showed that BIECs from fetal intestine maintained an in vivo phenotype as reflected in cytokeratin expression, expression of antigens restricted to intestinal enterocytes, and induced interleukin-8 (IL-8) production. BIECs could be infected by and support replication of bovine rotavirus. A semi-high-throughput enzyme-linked immunosorbent assay-based assay that measured IL-8 production by BIECs was established and used to screen commercially available natural compounds for novel adjuvant activity. Five novel hits were identified, demonstrating the utility of the assay for selecting and screening new epithelial cell adjuvants. Although the identified compounds had not previously been shown to induce IL-8 production in epithelial cells, other known functions for 3 of the 5 were consistent with this activity. Statistical analysis of the throughput data demonstrated that the assay is adaptable to a high-throughput format for screening both synthetic and natural product derived compound libraries.
Key Words: innate adjuvant • epithelial cell • cytokine • mucosa
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