Functional & Integrative Genomics

, Volume 10, Issue 1, pp 53–61 | Cite as

Greater expression of TLR2, TLR4, and IL6 due to negative energy balance is associated with lower expression of HLA-DRA and HLA-A in bovine blood neutrophils after intramammary mastitis challenge with Streptococcus uberis

  • Kasey M. Moyes
  • James K. Drackley
  • Dawn E. Morin
  • Juan J. Loor
Original Paper


Our objectives were to compare gene expression profiles in blood polymorphonuclear cells (PMN) during a Streptococcus uberis intramammary challenge between lactating cows subjected to feed restriction to induce negative energy balance (NEB; n = 5) and cows fed ad libitum to maintain positive energy balance (PEB; n = 5). After 5 days of feed restriction, one rear mammary quarter of each cow was inoculated with 5,000 cfu of S. uberis. Blood PMN were isolated at 24 h post-inoculation from all cows for mRNA expression via quantitative polymerase chain reaction for 20 genes associated with immune response and metabolism. A total of 12 genes were differentially expressed in blood PMN in NEB versus PEB cows. Upregulated genes by NEB were ALOX5AP, CPNE3, IL1R2, IL6, TLR2, TLR4, and THY1, and downregulated genes were HLA-DRA, HLA-A, IRAK1, SOD1, and TNF. Network analysis revealed that TNF was associated with several of the affected genes in NEB cows compared with PEB cows. Results showed that 24 h after intramammary challenge with S. uberis, cows in NEB had altered PMN expression of genes involved with immune response. Our data provide new information on transcriptomic mechanisms associated with NEB and the corresponding inhibition of immune response in lactating dairy cows.


Cattle Neutrophil Energy balance Streptococcus uberis 

Supplementary material

10142_2009_154_MOESM1_ESM.doc (244 kb)
ESM 1The file contains additional materials and methods. RNA isolation; data mining using IPA Knowledge Base (Ingenuity Systems, Inc.); primer design and testing for quantitative polymerase chain reaction (qPCR); identification of internal controls accompanied by four tables and one figure, which include Online Resource Table 1: GenBank accession number, hybridization position, sequence, amplicon size, and source of primers for Bos taurus used to analyze gene expression by qPCR. Online Resource Table 2: sequencing results obtained from qPCR product of Bos taurus specific primers. Online Resource Table 3: sequencing results of genes using BLASTN ( from NCBI against nucleotide collection with total score. Online Resource Table 4: slope and coefficient of determination of the standard curve (R2), efficiency (E)2, and median cycle threshold (Ct) of the measured transcripts. Online Resource Figure 1: stability (M) of gene expression ratios of potential ICG using geNorm software (a). geNorm analysis of optimal number of ICG for qPCR normalization for a subset of genes differentially expressed on microarray (b). (DOC 243 kb)


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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Kasey M. Moyes
    • 1
    • 3
    • 5
  • James K. Drackley
    • 1
    • 4
  • Dawn E. Morin
    • 2
    • 4
  • Juan J. Loor
    • 1
    • 3
  1. 1.Department of Animal SciencesUniversity of IllinoisUrbanaUSA
  2. 2.College of Veterinary MedicineUniversity of IllinoisUrbanaUSA
  3. 3.Mammalian NutriPhysioGenomicsUniversity of IllinoisUrbanaUSA
  4. 4.University of IllinoisUrbanaUSA
  5. 5.Faculty of Agricultural SciencesAarhus UniversityTjeleDenmark

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