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Expression of genes associated with inflammation induced by ex vivo exposure to lipopolysaccharide in peripheral blood leukocytes from horses with gastrointestinal disease

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  • 1 Departments of Large Animal Medicines, College of Veterinary Medicine, University of Georgia, Athens, GA 30602.
  • | 2 Departments of Large Animal Medicines, College of Veterinary Medicine, University of Georgia, Athens, GA 30602.
  • | 3 Departments of Large Animal Medicines, College of Veterinary Medicine, University of Georgia, Athens, GA 30602.
  • | 4 Population Health, College of Veterinary Medicine, University of Georgia, Athens, GA 30602.
  • | 5 Population Health, College of Veterinary Medicine, University of Georgia, Athens, GA 30602.
  • | 6 Physiology and Pharmacology, College of Veterinary Medicine, University of Georgia, Athens, GA 30602.

Abstract

Objective—To investigate the effect of ex vivo exposure to lipopolysaccharide (LPS) on the expression of inflammatory genes in leukocytes from horses with gastrointestinal (Gl) disease and determine whether the pattern or magnitude of the response to LPS correlated with the type of disease and outcome.

Animals—49 horses with Gl disease and 10 healthy horses

Procedures—Leukocytes were isolated from blood samples and submitted to 3 protocols: immediate freezing, freezing after 4-hour incubation in medium, and freezing after 4-hour incubation in medium containing LPS. Expression of 14 genes associated with inflammation was assessed via PCR assay. Results were compared by disease type and outcome

Results—Horses with Gl disease had colic of unknown etiology (n = 8), Gl inflammation or strangulation (18), or nonstrangulating Gl obstruction (23). Among the 44 horses receiving treatment, 38 were discharged from the hospital and 6 died or were euthanized. Incubation of leukocytes in medium alone changed the expression of several genes. Incubation with LPS resulted in increased expression of interleukin-10 and monocyte chemotactic protein-3 in leukocytes from healthy and sick horses. Leukocytes from horses with nonstrangulating obstruction and horses that survived had less pronounced LPS-induced increases in interleukin-10 expression than did cells from healthy horses. The opposite was evident for monocyte chemotactic protein-3.

Conclusions and Clinical Relevance—No evidence existed for a reduced response of leukocytes from horses with gastrointestinal disease to ex vivo exposure to LPS. Leukocyte expression of inflammatory genes after ex vivo incubation with LPS appeared to be related to pathogenesis and prognosis. (Am J Vet Res 2010;71:1162—1169)

Abstract

Objective—To investigate the effect of ex vivo exposure to lipopolysaccharide (LPS) on the expression of inflammatory genes in leukocytes from horses with gastrointestinal (Gl) disease and determine whether the pattern or magnitude of the response to LPS correlated with the type of disease and outcome.

Animals—49 horses with Gl disease and 10 healthy horses

Procedures—Leukocytes were isolated from blood samples and submitted to 3 protocols: immediate freezing, freezing after 4-hour incubation in medium, and freezing after 4-hour incubation in medium containing LPS. Expression of 14 genes associated with inflammation was assessed via PCR assay. Results were compared by disease type and outcome

Results—Horses with Gl disease had colic of unknown etiology (n = 8), Gl inflammation or strangulation (18), or nonstrangulating Gl obstruction (23). Among the 44 horses receiving treatment, 38 were discharged from the hospital and 6 died or were euthanized. Incubation of leukocytes in medium alone changed the expression of several genes. Incubation with LPS resulted in increased expression of interleukin-10 and monocyte chemotactic protein-3 in leukocytes from healthy and sick horses. Leukocytes from horses with nonstrangulating obstruction and horses that survived had less pronounced LPS-induced increases in interleukin-10 expression than did cells from healthy horses. The opposite was evident for monocyte chemotactic protein-3.

Conclusions and Clinical Relevance—No evidence existed for a reduced response of leukocytes from horses with gastrointestinal disease to ex vivo exposure to LPS. Leukocyte expression of inflammatory genes after ex vivo incubation with LPS appeared to be related to pathogenesis and prognosis. (Am J Vet Res 2010;71:1162—1169)

Contributor Notes

Supported by the Morris Animal Foundation and the White Fox Farm Research Fund.

Address correspondence to Dr. Lopes (maflopes@gmail.com).