Expression and distribution of antimicrobial peptides in the skin of healthy beagles
Tóm tắt
Antimicrobial peptides (AMPs) are small proteins involved in defense against pathogenic organisms. Defensins and cathelicidin are the most frequently studied human AMPs. Our goals were to determine the distribution of AMPs and evaluate their mRNA expression in normal canine skin. Skin biopsies were taken from six healthy beagles. The relative transcript level of canine cathelicidin (cCath) and β‐defensin (cBD)‐1, cBD2 and cBD3 mRNA was quantified using quantitative real‐time polymerase chain reaction. Indirect immunofluorescence (IIF), using polyclonal antibodies against cBD2, cBD3 and cCath, was used to evaluate protein localization in the skin of healthy dogs. The Pfaffl method, using experimentally determined primer efficiencies of amplification, was used to determine the expression level of cCath, cBD1 and cDB3 relative to cDB2. The levels of cCath, cBD3 and cBD1 mRNA were 358, 296 and 177 times higher than those of cBD2, respectively. Using IIF, cBD2 and cBD3 protein fluorescence was detected in all layers of the epidermis, whereas cCath was detected predominantly in the stratum granulosum and corneum. In addition, antimicrobial peptide detection was limited to the infundibular portion of the pilosebaceous units. We have validated useful methods to evaluate AMPs in canine skin. Further studies are needed to compare AMP expression in healthy dogs with that of dogs with inflammatory skin conditions.
Từ khóa
Tài liệu tham khảo
Agerberth B, 2000, The human antimicrobial and chemotactic peptides LL‐37 and α‐defensins are expressed by specific lymphocyte and monocytes populations, Blood, 96, 3086, 10.1182/blood.V96.9.3086
User Bulletin Applied Biosystems 75000/75000 Fast Real‐Time PCR Systems. Available at:http://www3.appliedbiosystems.com/cms/groups/mcb_support/documents/generaldocuments/cms_050637.pdf[accessed 13 June 2009].
Suttmann H, 2008, Antimicrobial peptides of the Cecropin‐family show potent antitumor activity against bladder cancer cells, BMC Urology, 3, 1
Bick RJ, 2007, Nuclear localization of HBD‐1 in human keratinocytes, Journal of Burns and Wounds, 7, 25
Poindexter BJ, 2005, Immunofluorescence deconvolution microscopy and image reconstruction of human defensins in normal and burned skin, Journal of Burns and Wounds, 4, 101