Expression of Vascular Cell Adhesion Molecule 1 (VCAM-1) in the Mammary Lymph Nodes of Cows with Subclinical Mastitis

Overview

Journal J Vet Res

Publisher Sciendo

Specialty Veterinary Medicine

Date 2018 Jul 7

PMID 29978074

Authors

Yuanyuan Chen

Wei Yang

Chuang Xu

Affiliations

Soon will be listed here.

Abstract

Introduction: Vascular cell adhesion molecule 1 (VCAM-1) is a member of Ig superfamily. The aim of this study was to prepare highly specific polyclonal antibodies against bovine VCAM-1 and to evaluate the expression of VCAM-1 in the mammary lymph nodes of cows with subclinical mastitis.

Material And Methods: The VCAM-1 gene was cloned from bovine Peyer's patches and inserted into the pGEX-4T-1 and pET-28a vectors. The recombinant plasmids pGEX-4T-1/VCAM-1 and pET-28a/VCAM-1 were transferred into BL21 and the recombinant strains were induced by isopropyl-D-thiogalactoside to produce fusion proteins tagged with polyhistidine (His) and glutathione S-transferase (GST), respectively. The expressed fusion proteins His-VCAM-1 and GST-VCAM-1 were identified by SDS-PAGE and Western blot. His-VCAM-1 protein was used as an antigen to immunise Wistar rats and polyclonal antibody serum against VCAM-1 was obtained.

Results: The serum titre tested by indirect ELISA was 128,000 using GST-VCAM-1 as the well coating antigen. Western blots indicated that the antibody recognised recombinant VCAM-1 protein as well as endogenous VCAM-1. In addition, using qPCR and Western blot, VCAM-1 mRNA and protein expression levels were measured in dairy cows with subclinical mastitis. It was demonstrated that VCAM-1 levels in the mammary lymph nodes of the cows were significantly higher than those from healthy controls (P < 0.05).

Conclusion: These results are to our knowledge the first report that VCAM-1 expression in the mammary lymph nodes is elevated in dairy cows with subclinical mastitis.

References

Peltzer J, Montespan F, Thepenier C, Boutin L, Uzan G, Rouas-Freiss N . Heterogeneous functions of perinatal mesenchymal stromal cells require a preselection before their banking for clinical use. Stem Cells Dev. 2014; 24(3):329-44. DOI: 10.1089/scd.2014.0327. View

Wedemeyer J, Vosskuhl K . Role of gastrointestinal eosinophils in inflammatory bowel disease and intestinal tumours. Best Pract Res Clin Gastroenterol. 2008; 22(3):537-49. DOI: 10.1016/j.bpg.2007.12.001. View

Clahsen T, Pabst O, Tenbrock K, Schippers A, Wagner N . Localization of dendritic cells in the gut epithelium requires MAdCAM-1. Clin Immunol. 2014; 156(1):74-84. DOI: 10.1016/j.clim.2014.11.005. View

Walsh G, Symon F, Lazarovils A, Wardlaw A . Integrin alpha 4 beta 7 mediates human eosinophil interaction with MAdCAM-1, VCAM-1 and fibronectin. Immunology. 1996; 89(1):112-9. PMC: 1456673. DOI: 10.1046/j.1365-2567.1996.d01-713.x. View

Jakubowski A, Rosa M, Bixler S, LOBB R, Burkly L . Vascular cell adhesion molecule (VCAM)-Ig fusion protein defines distinct affinity states of the very late antigen-4 (VLA-4) receptor. Cell Adhes Commun. 1995; 3(2):131-42. DOI: 10.3109/15419069509081282. View

Liao B, Tang Y, Hu F, Zhou W, Yao X, Hong W . Serum levels of soluble vascular cell adhesion molecules may correlate with the severity of dengue virus-1 infection in adults. Emerg Microbes Infect. 2015; 4(4):e24. PMC: 4575393. DOI: 10.1038/emi.2015.24. View

Wang M, Sugimoto M, Ikeda S, Kume S . Effects of coumestrol administration to maternal mice during pregnancy and lactation on immunoblogulin A-secreting cells in mammary glands. Anim Sci J. 2013; 84(4):322-7. DOI: 10.1111/asj.12007. View

Kunkel E, Butcher E . Chemokines and the tissue-specific migration of lymphocytes. Immunity. 2002; 16(1):1-4. DOI: 10.1016/s1074-7613(01)00261-8. View

Hession C, Tizard R, Vassallo C, Schiffer S, Goff D, Moy P . Cloning of an alternate form of vascular cell adhesion molecule-1 (VCAM1). J Biol Chem. 1991; 266(11):6682-5. View

10.

Springer T . Traffic signals on endothelium for lymphocyte recirculation and leukocyte emigration. Annu Rev Physiol. 1995; 57:827-72. DOI: 10.1146/annurev.ph.57.030195.004143. View

11.

Ahrens R, Waddell A, Seidu L, Blanchard C, Carey R, Forbes E . Intestinal macrophage/epithelial cell-derived CCL11/eotaxin-1 mediates eosinophil recruitment and function in pediatric ulcerative colitis. J Immunol. 2008; 181(10):7390-9. PMC: 2728352. DOI: 10.4049/jimmunol.181.10.7390. View

12.

Cybulsky M, FRIES J, Williams A, Sultan P, Eddy R, Byers M . Gene structure, chromosomal location, and basis for alternative mRNA splicing of the human VCAM1 gene. Proc Natl Acad Sci U S A. 1991; 88(17):7859-63. PMC: 52403. DOI: 10.1073/pnas.88.17.7859. View

13.

Osborn L, Vassallo C, Benjamin C . Activated endothelium binds lymphocytes through a novel binding site in the alternately spliced domain of vascular cell adhesion molecule-1. J Exp Med. 1992; 176(1):99-107. PMC: 2119269. DOI: 10.1084/jem.176.1.99. View

14.

Suraud V, Olivier M, Bodier C, Guilloteau L . Differential expression of homing receptors and vascular addressins in tonsils and draining lymph nodes: Effect of Brucella infection in sheep. Vet Immunol Immunopathol. 2006; 115(3-4):239-50. DOI: 10.1016/j.vetimm.2006.11.008. View

15.

Polte T, Newman W, Gopal T . Full length vascular cell adhesion molecule 1 (VCAM-1). Nucleic Acids Res. 1990; 18(19):5901. PMC: 332350. DOI: 10.1093/nar/18.19.5901. View

16.

Bar D, Tauer L, Bennett G, Gonzalez R, Hertl J, Schukken Y . The cost of generic clinical mastitis in dairy cows as estimated by using dynamic programming. J Dairy Sci. 2008; 91(6):2205-14. DOI: 10.3168/jds.2007-0573. View

17.

Osborn L, Hession C, Tizard R, Vassallo C, Luhowskyj S, LOBB R . Direct expression cloning of vascular cell adhesion molecule 1, a cytokine-induced endothelial protein that binds to lymphocytes. Cell. 1989; 59(6):1203-11. DOI: 10.1016/0092-8674(89)90775-7. View

18.

Bai R, Bai H, Kuse M, Ideta A, Aoyagi Y, Fujiwara H . Involvement of VCAM1 in the bovine conceptus adhesion to the uterine endometrium. Reproduction. 2014; 148(2):119-27. DOI: 10.1530/REP-13-0655. View

19.

Carvalho A, Elia C, de Souza H, Elias P, Pontes E, Pitanga Lukashok H . Immunohistochemical study of intestinal eosinophils in inflammatory bowel disease. J Clin Gastroenterol. 2003; 36(2):120-5. DOI: 10.1097/00004836-200302000-00006. View

20.

Schaefer M, Sarkar S, Schwarz M, Friebe A . Soluble Intracellular Adhesion Molecule-1 in Patients with Unipolar or Bipolar Affective Disorders: Results from a Pilot Trial. Neuropsychobiology. 2016; 74(1):8-14. DOI: 10.1159/000446919. View