title: Structural and immunological effects of skin cryoablation in a mouse model.
abstract: Cryoablation is therapeutically applied for various disorders in several organs, and skin diseases are typical targets as this cryotherapy has been widely used for viral warts, benign tumors, and actinic keratosis. The main mechanisms of cryoablation consist of direct freezing effect on skin constituents, thrombosis formation in microcirculation, and subsequent immunological responses. Among them, however, the immunological mechanism remains unelucidated, and it is an issue how the direct freezing injury induces immunological consequences. We established a mouse cryoablation model with liquid nitrogen applied to the shaved back skin, and used this system to study the immunological excitement. After application of liquid nitrogen, the thermal decrease ratio was -25°C/sec or less and the lowest temperature was less than -100°C, which was sufficient to induce ulceration. Destruction of cornified layer and necrosis of epidermal cells were observed in transmission electron microscopy image, and increased transepidermal water loss and skin permeability were detected by the functional measurements. By flow cytometry, antigen-presenting dendritic cells (DCs), including PDCA1+B220+CD19- plasmacytoid DCs (pDCs) and CD11c+ myeloid DCs, as well as neutrophils and macrophages were increased in subcutaneous tissue. In parallel, the mRNA expressions of interferon α1 which are known as pDC-producing cytokines, was elevated. We also found marked degranulation of mast cells, providing a possibility that released histamine attracts pDCs. Finally, FITC migration assay revealed that pDCs and CD11c+ DCs emigrated from the cryoablated skin to the draining lymph nodes. Our study suggests that cryoablation induces destruction of the barrier/epidermis, accumulation of pDCs and CD11c+ DCs to the skin, and migration of DCs to regional lymph nodes. Viral elements or tumor cell lysates released from damaged keratinocytes may stimulate the DCs, thereby leading to antiviral or antitumor effect.
authors.affiliation: Department of Dermatology, Hamamatsu University School of Medicine, Hamamatsu, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Kasuya
authors.affiliation: Ultrastructural Morphology Laboratory, Research Equipment Centre, Hamamatsu University School of Medicine, Hamamatsu, Japan.
authors.forename: Isao
authors.initials: I
authors.lastname: Ohta
authors.affiliation: Department of Dermatology, Hamamatsu University School of Medicine, Hamamatsu, Japan.
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Tokura
mesh_terms: D000818:Animals; D017951:Antigen Presentation / Q000276:immunology; D039521:CD11c Antigen / Q000276:immunology; D003452:Cryosurgery / Q000009:adverse effects* / Q000379:methods; D003713:Dendritic Cells / Q000276:immunology / Q000473:pathology; D062109:Dermatologic Surgical Procedures; D004817:Epidermis / Q000276:immunology / Q000473:pathology; D005260:Female; D016898:Interferon-alpha / Q000276:immunology; D015603:Keratinocytes / Q000276:immunology / Q000473:pathology; D008198:Lymph Nodes / Q000276:immunology / Q000473:pathology; D008264:Macrophages / Q000276:immunology / Q000473:pathology; D008407:Mast Cells / Q000276:immunology / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D009504:Neutrophils / Q000276:immunology / Q000473:pathology; D012333:RNA, Messenger / Q000276:immunology; D012867:Skin / Q000276:immunology* / Q000473:pathology*
chemical_list: D039521:CD11c Antigen; D016898:Interferon-alpha; D012333:RNA, Messenger
references.citation: Ann N Y Acad Sci. 2012 Jun;1257:158-66
references.pmid: 22671602
references.citation: Cryobiology. 2009 Feb;58(1):1-11
references.pmid: 19007768
references.citation: J Immunol. 2010 Feb 15;184(4):2116-23
references.pmid: 20083654
references.citation: Nat Med. 1999 Nov;5(11):1249-55
references.pmid: 10545990
references.citation: Surgery. 1997 Oct;122(4):742-7
references.pmid: 9347851
references.citation: Int Immunol. 2012 Feb;24(2):107-16
references.pmid: 22232416
references.citation: Cryobiology. 1982 Jun;19(3):247-62
references.pmid: 7105777
references.citation: Cancer Res. 2006 Jul 15;66(14):7285-92
references.pmid: 16849578
references.citation: J Immunol. 2005 May 1;174(9):5224-32
references.pmid: 15843518
references.citation: J Exp Med. 2000 Feb 7;191(3):423-34
references.pmid: 10662788
references.citation: Arch Dermatol. 1977 Sep;113(9):1209-13
references.pmid: 900963
references.citation: Methods. 2001 Dec;25(4):402-8
references.pmid: 11846609
references.citation: PLoS One. 2011;6(6):e21251
references.pmid: 21698180
references.citation: Biochim Biophys Acta. 1953 Mar;10(3):414-26
references.pmid: 13058999
references.citation: J Urol. 1970 Jul;104(1):154-9
references.pmid: 4987666
references.citation: J Urol. 2010 Jan;183(1):333-8
references.pmid: 19914660
references.citation: Br J Pharmacol. 1980 Aug;69(4):615-23
references.pmid: 6108140
references.citation: J Urol. 1972 Dec;108(6):928-32
references.pmid: 4117196
references.citation: Cryobiology. 1998 Nov;37(3):171-86
references.pmid: 9787063
references.citation: Viruses. 2013 Nov;5(11):2624-42
references.pmid: 24169630
references.citation: Cancer Immunol Immunother. 2012 Aug;61(8):1279-88
references.pmid: 22294456
references.citation: J Invest Dermatol. 2011 Aug;131(8):1668-76
references.pmid: 21614010
references.citation: Nat Rev Cancer. 2014 Mar;14(3):199-208
references.pmid: 24561446
references.citation: BJU Int. 2005 Jun;95(9):1187-91
references.pmid: 15892798
references.citation: Curr Biol. 2001 Jul 24;11(14):R569-72
references.pmid: 11509257
references.citation: Urology. 1996 Sep;48(3):441-7
references.pmid: 8804499
references.citation: J Adhes Sci Technol. 2013 Feb 1;27(3):227-243
references.pmid: 23543070
references.citation: J Virol. 2002 Oct;76(20):10401-16
references.pmid: 12239317
references.citation: J Clin Invest. 2004 Sep;114(5):729-38
references.pmid: 15343392
references.citation: Nat Rev Immunol. 2004 Jan;4(1):46-54
references.pmid: 14704767
references.citation: J Dermatol Sci. 2014 May;74(2):116-24
references.pmid: 24485663
references.citation: J Immunol. 2006 Sep 1;177(5):3260-5
references.pmid: 16920966
references.citation: Cryobiology. 2011 Aug;63(1):38-45
references.pmid: 21536022