Construction and validation of a Bovine Innate Immune Microarray
Open Access
- 22 September 2005
- journal article
- Published by Springer Science and Business Media LLC in BMC Genomics
- Vol. 6 (1), 135
- https://doi.org/10.1186/1471-2164-6-135
Abstract
Background: Microarray transcript profiling has the potential to illuminate the molecular processes that are involved in the responses of cattle to disease challenges. This knowledge may allow the development of strategies that exploit these genes to enhance resistance to disease in an individual or animal population.Results: The Bovine Innate Immune Microarray developed in this study consists of 1480 characterised genes identified by literature searches, 31 positive and negative control elements and 5376 cDNAs derived from subtracted and normalised libraries. The cDNA libraries were produced from 'challenged' bovine epithelial and leukocyte cells. The microarray was found to have a limit of detection of 1 pg/μg of total RNA and a mean slide-to-slide correlation co-efficient of 0.88. The profiles of differentially expressed genes from Concanavalin A (ConA) stimulated bovine peripheral blood lymphocytes were determined. Three distinct profiles highlighted 19 genes that were rapidly up-regulated within 30 minutes and returned to basal levels by 24 h; 76 genes that were up-regulated between 2–8 hours and sustained high levels of expression until 24 h and 10 genes that were down-regulated. Quantitative real-time RT-PCR on selected genes was used to confirm the results from the microarray analysis. The results indicate that there is a dynamic process involving gene activation and regulatory mechanisms re-establishing homeostasis in the ConA activated lymphocytes. The Bovine Innate Immune Microarray was also used to determine the cross-species hybridisation capabilities of an ovine PBL sample.Conclusion: The Bovine Innate Immune Microarray has been developed which contains a set of well-characterised genes and anonymous cDNAs from a number of different bovine cell types. The microarray can be used to determine the gene expression profiles underlying innate immune responses in cattle and sheep.This publication has 72 references indexed in Scilit:
- Johne's disease in cattle is associated with enhanced expression of genes encoding IL-5, GATA-3, tissue inhibitors of matrix metalloproteinases 1 and 2, and factors promoting apoptosis in peripheral blood mononuclear cellsVeterinary Immunology and Immunopathology, 2005
- A 7872 cDNA microarray and its use in bovine functional genomicsVeterinary Immunology and Immunopathology, 2005
- Construction and application of a bovine immune-endocrine cDNA microarrayVeterinary Immunology and Immunopathology, 2004
- T-cell-receptor- and B-cell-receptor-mediated activation of NF-κB in lymphocytesCurrent Opinion in Immunology, 2004
- Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔCT MethodMethods, 2001
- Sequence Evaluation of Four Pooled-Tissue Normalized Bovine cDNA Libraries and Construction of a Gene Index for CattleGenome Research, 2001
- Analysis of Variance for Gene Expression Microarray DataJournal of Computational Biology, 2000
- Identification of a new member of the tumor necrosis factor family and its receptor, a human ortholog of mouse GITRCurrent Biology, 1999
- Gapped BLAST and PSI-BLAST: a new generation of protein database search programsNucleic Acids Research, 1997
- Immortalization and characterization of bovine peritoneal macrophages transfected with SV40 plasmid DNAVeterinary Immunology and Immunopathology, 1995