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Detection of calprotectin and its correlation to the accumulation of neutrophils within equine large colon during ischaemia and reperfusion.
Abstract: The cytosolic protein complex, calprotectin, is abundant in neutrophils and could be used to improve the ability to localise and assess neutrophil infiltration in the equine intestine during ischaemia and reperfusion (I/R), but further study is required. Objective: To assess the number of calprotectin-containing cells by immunohistochemistry in correlation with direct counting and scoring of neutrophils in the equine colon during I/R. Methods: One and 2 h ischaemia of the left dorsal colon were induced, followed by 30 min reperfusion under general anaesthesia or by 18 h reperfusion after anaesthetic recovery. Biopsies were processed for light microscopy and stained with H/E for detection of neutrophils. To identify calprotectin-containing cells, immunohistochemistry was performed on formalin-fixed tissues with the murine MAC 387 antibody and a biotin-free peroxidase staining procedure. The number of neutrophils within submucosal venules and the colonic mucosa were calculated and compared with the number of calprotectin-positive cells. Results: The number of calprotectin-positive cells within submucosal venules and within the colonic mucosa correlated significantly with the accumulation of neutrophils within the corresponding tissue segments. Within the submucosal venules, both calprotectin-positive cells and H/E-stained neutrophils increased with duration of ischaemia and peaked after 30 min of reperfusion. After 18 h reperfusion the number of these cells declined within the vessels. After 2 h ischaemia, neutrophils started to migrate into the mucosa towards the epithelium, with a significant increase over time during reperfusion, and peak infiltration after 18 h reperfusion. Conclusions: Neutrophil infiltration into the colon after I/R is a time-dependent process, involving migration through the submucosa towards the epithelium.
Publication Date: 2008-05-20 PubMed ID: 18487110DOI: 10.2746/042516408X302500Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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The research investigates how calprotectin, a protein complex found in neutrophils, could aid in detecting and assessing neutrophil infiltration during instances of ischaemia and reperfusion (I/R) in the equine intestine. Through a series of experiments, it was found that the number of calprotectin-positive cells significantly correlated with the accumulation of neutrophils.
Methods
- The research induced one and two hour ischaemia in the left dorsal colon of horses, which was followed by 30 minutes of reperfusion under general anaesthesia or 18 hours of reperfusion after recovery from anaesthesia.
- Biopsies of the affected tissues were taken and processed for light microscopy to check for the presence of neutrophils. The tissue samples were stained with Hematoxylin and Eosin (H/E) and immunohistochemistry was performed on formalin-fixed tissues using the MAC 387 antibody and biotin-free peroxidase staining procedure to identify calprotectin-positive cells.
- The researchers then calculated the number of neutrophils in the submucosal venules and the colonic mucosa and compared that with the number of calprotectin-positive cells.
Results
- The study found that the number of calprotectin-positive cells within the submucosal venules and the colonic mucosa significantly correlated with the accumulation of neutrophils in those areas.
- Both calprotectin-positive cells and H/E-stained neutrophils increased with the duration of ischaemia and peaked after 30 minutes of reperfusion.
- After the horses were allowed to recover from the anaesthesia for 18 hours, the number of calprotectin-positive cells and H/E-stained neutrophils declined within the blood vessels.
- After two hours of ischaemia, the neutrophils started to migrate into the mucosa towards the epithelium, peaking after 18 hours of reperfusion.
Conclusions
- The study concludes that neutrophil infiltration into the colon following ischaemia and reperfusion is a time-dependent process, and involves migration through the submucosa towards the epithelium.
- Calprotectin, being abundant in neutrophils, could therefore serve as a potential marker for assessing the occurrence and severity of neutrophil infiltration during instances of ischaemia and reperfusion in the equine intestine.
Cite This Article
APA
Grosche A, Morton AJ, Polyak MM, Matyjaszek S, Freeman DE.
(2008).
Detection of calprotectin and its correlation to the accumulation of neutrophils within equine large colon during ischaemia and reperfusion.
Equine Vet J, 40(4), 393-399.
https://doi.org/10.2746/042516408X302500 Publication
Researcher Affiliations
- Island Whirl Equine Colic Research Laboratory, Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32610, USA.
MeSH Terms
- Animals
- Colon / blood supply
- Colon / immunology
- Horse Diseases / immunology
- Horse Diseases / pathology
- Horses
- Immunohistochemistry / veterinary
- Leukocyte Count / veterinary
- Leukocyte L1 Antigen Complex / immunology
- Leukocyte L1 Antigen Complex / isolation & purification
- Neutrophils / metabolism
- Neutrophils / physiology
- Reperfusion Injury / immunology
- Reperfusion Injury / pathology
- Reperfusion Injury / veterinary
- Time Factors
Citations
This article has been cited 5 times.- Muñoz-Prieto A, Contreras-Aguilar MD, Cerón JJ, Ayala de la Peña I, Martín-Cuervo M, Eckersall PD, Holm Henriksen IM, Tecles F, Hansen S. Changes in Calprotectin (S100A8-A9) and Aldolase in the Saliva of Horses with Equine Gastric Ulcer Syndrome. Animals (Basel) 2023 Apr 16;13(8).
- Jana S, Mitra P, Roy S. Proficient Novel Biomarkers Guide Early Detection of Acute Kidney Injury: A Review. Diseases 2022 Dec 30;11(1).
- König KS, Verhaar N, Hopster K, Pfarrer C, Neudeck S, Rohn K, Kästner SBR. Ischaemic preconditioning and pharmacological preconditioning with dexmedetomidine in an equine model of small intestinal ischaemia-reperfusion. PLoS One 2020;15(4):e0224720.
- Carossino M, Loynachan AT, Canisso IF, Cook RF, Campos JR, Nam B, Go YY, Squires EL, Troedsson MHT, Swerczek T, Del Piero F, Bailey E, Timoney PJ, Balasuriya UBR. Equine Arteritis Virus Has Specific Tropism for Stromal Cells and CD8(+) T and CD21(+) B Lymphocytes but Not for Glandular Epithelium at the Primary Site of Persistent Infection in the Stallion Reproductive Tract. J Virol 2017 Jul 1;91(13).
- Schnabel CL, Steinig P, Koy M, Schuberth HJ, Juhls C, Oswald D, Wittig B, Willenbrock S, Murua Escobar H, Pfarrer C, Wagner B, Jaehnig P, Moritz A, Feige K, Cavalleri JM. Immune response of healthy horses to DNA constructs formulated with a cationic lipid transfection reagent. BMC Vet Res 2015 Jun 23;11:140.
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