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. 2009 Jan;73(1):49-57.

Assessment of bovine mammary chemokine gene expression in response to lipopolysaccharide, lipotechoic acid + peptidoglycan, and CpG oligodeoxynucleotide 2135

Jeremy A Mount  1 , Niel A Karrow, Jeff L Caswell, Herman J Boermans, Ken E Leslie
Affiliations

Assessment of bovine mammary chemokine gene expression in response to lipopolysaccharide, lipotechoic acid + peptidoglycan, and CpG oligodeoxynucleotide 2135

Jeremy A Mount et al. Can J Vet Res. 2009 Jan.

Abstract
in English, French

During intramammary infections pathogen associated molecular patterns (PAMPs) induce an inflammatory response, recognized clinically as mastitis. Recognition of PAMPs by mammary cells leads to the production of the pro-inflammatory cytokines, TNF-alpha and IL-1beta. These cytokines augment the secretion of various chemokines that are responsible for directing the host cellular immune response, and consequently the outcome of infection. Previous research has shown that gram-negative and gram-positive bacteria elicit different types of innate immune responses. The purpose of this study, therefore, was to characterize the expression of various chemokine genes in bovine mammary gland explants in response to lipopolysaccharide (LPS), peptidoglycan (PTG) combined with lipotechoic acid (LTA), and CpG oligodeoxynucleotide (CpG-ODN) 2135 representing gram-negative bacteria, gram-positive bacteria, and bacterial DNA, respectively, to determine if these PAMPs induce different chemokine gene expression patterns. Explants from 3 Holstein cows were cultured with 10 microg/mL of LPS, LTA + PTG, or CpG-ODN 2135 for 6 and 24 h. Total RNA was extracted and the expression of CXCL8, MCP-1, MCP-2, MCP-3, MIP1-alpha, and RANTES genes was measured by real-time polymerase chain reaction (RT-PCR). Lipopolysaccharide significantly induced MCP-1, MCP-2, and MCP-3 expression, and slightly increased CXCL8 gene expression. The combined PAMPs, LTA + PTG, on the other hand, significantly induced MCP-1 gene expression, and slightly increased MCP-3 expression. No significant expression differences for any of the chemokine genes were observed in explants stimulated with CpG-ODN 2135. These results demonstrate that PAMPs associated with different mastitis-causing pathogens induce chemokine-specific gene expression patterns that may contribute to different innate immune responses to bacteria.

Lors d’infections intra-mammaires les patrons moléculaires associés aux agents pathogènes (PAMPs) induisent une réponse inflammatoire reconnue cliniquement comme une mammite. La reconnaissance des PAMPs par les cellules de la glande mammaire entraîne la production des cytokines pro-inflammatoires TNF-α et IL-1β. Ces cytokines augmentent la sécrétion de diverses chimiokines qui sont responsables d’orienter la réponse immunitaire cellulaire de l’hôte, et conséquemment l’issue de l’infection. Les recherches antérieures ont démontré que les bactéries à Gram négatif et à Gram positif induisent différents types de réponses immunes innées. Ainsi, l’objectif de cette étude était de caractériser l’expression de différents gènes de chimiokines dans des explants de glande mammaire en réponse au lipolysaccharide (LPS), peptidoglycan (PTG) combiné avec de l’acide lipotéchoique (LTA), et CpG oligodéoxynucléotide (CpG-ODN) 2135 représentant respectivement les bactéries à Gram négatif, les bactéries à Gram positif et l’ADN bactérien, afin de déterminer si ces PAMPs induisent différents patrons d’expression des gènes des chimiokines. Des explants provenant de trois vaches Holstein ont été cultivées avec 10 μg/mL de LPS, LTA + PTG ou CpG-ODN 2135 pour 6 et 24 heures. L’ARN total a été extrait et l’expression des gènes de CXCL8, MCP-1, MCP-2, MCP-3, M1P1-α et RANTES mesurée par RT-PCR. Le LPS a induit l’expression significative de MCP-1, MCP-2 et MCP-3, ainsi qu’une légère augmentation de l’expression du gène de CXCL8. D’autre part, le PAMPs combiné LTA + PTG a induit une expression significative du gène de MCP-1, et une légère augmentation de l’expression de MCP-3. Dans les explants stimulés avec CpG-ODN 2135 aucune différence significative d’expression n’a été observée pour tous les gènes de chimiokines. Ces résultats démontrent que les PAMPs associés avec les agents pathogènes causant la mammite induisent des patrons d’expression des gènes spécifiques des chimiokines qui peuvent contribuer à différentes réponses immunitaires innées envers les bactéries.

(Traduit par Docteur Serge Messier)

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Figures

Figure 1
Figure 1
Control mammary explants cultured for 6 h (A and B) and 24 h (C and D) during PAMP exposure. The explants were stained with either hematoxylin and eosin (A and C) to stain dark nuclear material and light cytoplasmic material, or vimentin (B and D) to stain cells of mesenchymal origin. These images are magnified 40 ×
Figure 2
Figure 2
Chemokine (CXCL8, RANTES, MIP1-α, MCP-1–3) and housekeeping gene expression (GAPDH, RPLPO) following mammary explant culture with LPS, LTA + PTG and CpG-ODN 2135. Since there was no significant treatment by time interaction, these results represent the average least squares means +/− the standard error of the 6- and 24-h sampling time points. Levels of significance of the treatment groups compared to the control are indicated by * = P < 0.10, ** = P < 0.05, and *** = P < 0.01.
See this image and copyright information in PMC

References

    1. Miller GY, Bartlett PC, Lance SE, Anderson J, Heider LE. Costs of clinical mastitis and mastitis prevention in dairy herds. J Am Vet Med Assoc. 1993;202:1230–1236. - PubMed
    1. Seegers H, Fourichon C, Beaudeau F. Production effects related to mastitis and mastitis economics in dairy cattle herds. Vet Res. 2003;34:475–491. - PubMed
    1. Fitzgerald JR, Hartigan PJ, Meaney WJ, Smyth CJ. Molecular population and virulence factor analysis of Staphylococcus aureus from bovine intramammary infection. J Appl Microbiol. 2000;88:1028–1037. - PubMed
    1. Miles H, Lesser W, Sears P. The economic implications of bioengineered mastitis control. J Dairy Sci. 1992;75:596–605. - PubMed
    1. Bannerman DD, Paape MJ, Goff JP, Kimura K, Lippolis JD, Hope JC. Innate immune response to intramammary infection with Serratia marcescens and Streptococcus uberis. Vet Res. 2004;35:681–700. - PubMed

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