Journal of Dermatological Science 66 (2012) 71–84

Letter to the Editor

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Journal of Dermatological Science

jou r nal h o mep ag e: w ww .e lsev ier . co m / jds

Ultraviolet irradiation modulates ABO blood group antigens inhuman skin in vivo: Possible implication in skin aging

The ABO blood group antigens (ABH antigens) are carbohydrateantigens, which are synthesized by stepwise addition of carbohy-drate chains. In cells with type-2 chains (Galb1–4GlcNAcb1–)such as keratinocytes, common precursors attached to lipids orproteins are synthesized by the orderly action of b-1,4-galacto-syltransferase (B4GALT)6, b-1,3-N-acetylglucosaminyltransferase(B3GNT)5, and B4GALT1,2,3, and 4. Then, H antigen is synthesizedby fucosyltransferase (FUT)1 via addition of L-fucose. In blood typeA or B, subsequent addition of N-acetyl-D-galactosamine or D-galactose by A or B transferase, polymorphisms of ABO gene havingonly four different amino acids, results in A or B antigen,

Fig. 1. ABH antigen expression in intrinsically aged and photoaged human skin. Skin sam

healthy young (mean age 30.8 � 8.2 years, n = 9) and elderly (72.6 � 6.8 years, n = 5) Korea

facial skin in young and elderly subjects with blood group B (hematoxylin–eosin, original m

same trend was observed for the expression of ABH antigen in those with blood group A

determined using a 5-point scale (0, no positive staining; 1, weak staining including focal po

very intense staining of large areas) by four dermatologists in a blinded manner. Error ba

0923-1811/$36.00 � 2012 Japanese Society for Investigative Dermatology. Published b

respectively (Supplementary Fig. 1). ABH antigens are expressedin diverse human tissues, including epithelial cells which linegastrointestinal, bronchopulmonary, genitourinary tracts, con-junctiva, and skin [1].

Differential pathogen adherence and interactions in associa-tion with ABO blood groups have been reported in severalmicroorganisms including Helicobacter pylori [2]. Gain or loss ofABH antigens in cancer cells may influence tumor behavior byaltering signal transduction, motility, resistance to apoptosis, andimmunosurveillance [2]. Recent genome-wide association stud-ies have identified associations between ABO genotype and levelsof soluble E-/P-selectin, intercellular adhesion molecule-1 (ICAM-1), and tumor necrosis factor-a(TNF-a) [3]. To date, various skindiseases, ranging from benign conditions like atopic dermatitis to

ples were obtained both from the buttock (sun-protected) and face (sun-exposed) of

n subjects. (a) Expression of B and H antigen of sun-protected buttock and sun-exposed

agnification �200). Insert, higher magnification of outlined portion in epithelium. The

, AB, and O (data not shown). (b) The mean staining intensity of each specimen was

sitively stained areas; 2, moderate staining; 3, strong staining with expanded areas; 4,

rs denote standard errors of the means. *P < 0.05. ns, not significant.

y Elsevier Ireland Ltd. All rights reserved.

Letters to the Editor / Journal of Dermatological Science 66 (2012) 71–8472

nonmelanoma skin cancers, have been reported to have anassociation with ABO genotypes [4], but experimental evidencesupporting these observations has remained scarce.

To investigate changes in ABH antigens associated with skinaging, skin samples were obtained both from the buttock (sun-protected) and face (sun-exposed) of healthy young (mean age30.8 � 8.2 years, n = 9) and elderly (72.6 � 6.8 years, n = 5) Koreansubjects. Formalin-fixed paraffin-embedded samples were stainedusing monoclonal anti-A, anti-B, or anti-H (type 2) antibody (1:100,Santa Cruz Biotechnology) and biotinylated secondary antibody aspreviously described [5]. The staining intensity was graded using a 5-point scale by four dermatologists in a blinded manner. Epidermis ofblood type A, B and AB subjects expressed corresponding A, B andboth A and B antigens, respectively, mainly along stratum granulo-sum and upper stratum spinosum as well as H antigen predominantlyin upper stratum spinosum (Supplementary Fig. 2). The skin of blood

Fig. 2. Ultraviolet (UV)-induced changes in ABH antigen expression in human skin in viv

phototherapy device, including F75/85W/UV21 fluorescent sunlamp (emission spectrum

The distribution of UV irradiation from the device was 0.5% UVC (below 280 nm), 56.7% U

Kodacel filter (TA401/407, Kodak), the irradiance of UV irradiation at the skin surface wa

measured at 30 cm (bulb-to-target distance) from the bulb was 1.0 mW/cm2. The buttock

erythema dose (MED) was determined at 24 h after irradiation. The MED ranges were 70

skin was irradiated with 2 MED of UV and UV-irradiated skin were obtained at 24, 48, a

representative figures among 13 volunteers (a; blood group A, b; blood group O subject) (

B and AB showed a similar pattern (data not shown). (c) Levels of ABO transferase, fuco

B4GALT4, b-1,3-N-acetylglucosaminyltransferase (B3GNT)5, and B4GALT6 mRNA were a

bars denote standard errors of the means. *P < 0.05 vs unirradiated control skin.

type O individuals exhibited stronger H antigen expression mostly instratum granulosum as well as stratum spinosum. These resultsconfirmed that ABH antigen is expressed in the outermost layers ofthe epidermis [1]. Sun-protected buttock skin exhibited similarpatterns of expression of ABH antigens regardless of age groups(Fig. 1a and b). In both young and elderly individuals, A or B antigen insun-exposed skin showed significantly weaker expression than insun-protected skin (P = 0.0183 by Wilcoxon signed-rank test). Hantigen showed slightly stronger, albeit not significant, expressionalong spinous layers in sun-exposed skin, compared with sun-protected skin (P = 0.380). Further stratification by each blood grouprevealed that patterns of ABH antigen expression were similar acrossdifferent ABO blood groups (data not shown).

Since expression of A or B antigen is decreased in sun-exposedskin, we next examined expression of ABH antigens followingacute UV irradiation. Buttock skin of 13 healthy subjects (blood

o. For the acute UV irradiation study, a Waldmann UV-800 (Waldmann, Germany)

between 275 and 380 nm, peak at 310–315 nm), was employed as the UV source.

VB (280–320 nm), and 42.8% UVA (320–400 nm) [5]. After filtering out UVC with a

s measured using a Waldmann UV meter (Model 585100). The mean UVB irradiance

skin of healthy volunteers (n = 13) was irradiated with filtered UV, and the minimal

–90 mJ/cm2 in Korean subjects with skin type III, IV, and V. Accordingly, the buttock

nd 72 h after UV irradiation, in addition to non-irradiated control skin. Images are

hematoxylin–eosin, original magnification: �200). Other subjects with blood group

syltransferase (FUT)1, b-1,4-galactosyltransferase (B4GALT)1, B4GALT2, B4GALT3,

nalyzed using real-time RT-PCR, and normalized with level of 36B4 transcripts. Error

Letters to the Editor / Journal of Dermatological Science 66 (2012) 71–84 73

group A, 3; B, 5; AB, 3; O, 2) was irradiated using a F75/85W/UV21UV device (Waldmann, Germany), and obtained by punch biopsy at0, 24, 48, and 72 h after 2 minimal erythema dose (MED) of UVirradiation. Of particular interest, expression of A, B, or H antigenalong stratum granulosum was strongly downregulated inirradiated skin as early as 24 h post-irradiation (Fig. 2a and b).In most specimens, expression of A, B, or H antigen along stratumgranulosum remained decreased until 72 h post-irradiation. Onthe contrary, H antigen expression showed marked upregulation inboth extent and intensity along the spinous and suprabasal layersfrom 48 h after UV irradiation. Notably, in blood group Oindividuals, strong H antigen normally present along stratumgranulosum decreased but its expression in deeper layersincreased. Glycosyltransferases involved in ABH antigen synthesiswere quantitated by real-time RT-PCR (CFX96, BIO-RAD). TotalRNA was prepared from separated epidermis using a TRIZOL(Invitrogen Life Technologies), and converted to cDNA using FirstStrand cDNA Synthesis Kit (Fermentas). ABO transferase mRNAshowed a substantial decrease after UV irradiation, with statisticalsignificance at 48 h. On the other hand, FUT1, B4GALT1, B4GALT2,B3GNT5, and B4GALT6 mRNA showed robust and significantincreases at 24 h post-irradiation and progressive declinesthereafter (Fig. 2c). These results suggest that the aforementioneddecrease in A or B antigen and increase in H antigen in UV-irradiated skin in vivo can be attributed to decreased mRNAexpression of ABO transferase and increased mRNA expression ofenzymes involved in synthesis of H antigen, respectively. TheInstitutional Review Board approved this study, and all subjectsprovided written informed consent.

To our knowledge, our result that UV-induced alterations ofABH antigens in epidermis are characterized by downregulation ofABH antigens along granular layers and upregulation of H antigenthroughout spinous layers is a novel finding, and ABH antigenalterations in human skin in vivo upon external stimuli, such as UV,have not yet been examined. UV-induced regulation of proteinglycosylation can be different depending on the type of glycosyla-tion [6], rather than a common phenomenon. In mucosal woundhealing processes, Lewisy and H antigens were upregulated [7], andLewisy-6/H-5-2 glycoconjugate (Ley/H) induced angiogenesis in ratcornea [8]. In addition, alteration of ABH antigens can result inactivity changes of glycoproteins, including epidermal growthfactor receptor (EGFR), FUT, von Willebrand factor (vWF), FactorVIII, and alkaline phosphatase [2]. In the absence of A antigen, EGFRshowed increases of receptor turnover, high affinity receptors,protein phosphorylation, and tyrosine kinase activity [9]. Theglycosylation status of vWF and Factor VIII influenced theirprocessing, resulting in their higher levels in O individuals [2].Moreover, inhibition of FUT1/4 decreased cell proliferation andtumor growth [10], suggesting that protein glycosylation canparticipate as a cause for changing cell properties. In this context,UV-induced alterations of ABO glycosylation in skin might affectmany physiological functions of various glycoproteins andglycolipids in photodamage.

Taken together, expression of ABO blood group antigensshowed distinct changes in response to acute UV irradiation, aswell as in chronically sun-exposed skin, suggesting that ABO bloodgroups might be implicated in acute UV-induced skin responsesand photoaging processes. Further investigations are warranted fordetermination of the potential mechanisms by which UVmodulates transcription of ABO blood group-associated glycosyl-transferases and its functional significance.

Acknowledgments

This work was supported by the Korea Science and EngineeringFoundation (KOSEF) grant funded by the Korea government (MEST)(No. 2009-0078847), the National Research Foundation of KoreaGrant funded by the Korean Government [NRF-2009-351-E00026],and a grant of the Korea Healthcare technology R&D Project,Ministry of Health & Welfare, Republic of Korea (Grant No.:A103017).

Appendix A. Supplementary data

Supplementary data associated with this article can be found, in

the online version, at doi:10.1016/j.jdermsci.2012.01.006.

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[9] Defize LH, Arndt-Jovin DJ, Jovin TM, Boonstra J, Meisenhelder J, Hunter T, et al.A431 cell variants lacking the blood group A antigen display increased highaffinity epidermal growth factor-receptor number, protein-tyrosine kinaseactivity, and receptor turnover. J Cell Biol 1988;107:939–49.

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Dong Hun Leea,b,c,1, Ji-Yong Junga,b,c,1, Jang-Hee Oha,b,c,Serah Leea,b,c, Yeon Kyung Kima,b,c, Jin Ho Chunga,b,c,*aDepartment of Biomedical Sciences and Dermatology,

Seoul National University College of Medicine, Seoul,

Republic of KoreabLaboratory of Cutaneous Aging Research, Biomedical Research

Institute, Seoul National University Hospital, Seoul,

Republic of KoreacInstitute of Dermatological Science, Seoul National University,

Seoul, Republic of Korea

*Corresponding author at: Department of Dermatology, SeoulNational University Hospital, 101 Daehak-ro, Jongno-Gu,Seoul 110-744, Republic of Korea.Tel.: +82 2 2072 2414; fax: +82 2 742 7344E-mail address: jhchung@snu.ac.kr (J.H. Chung)

1These authors contributed equally to this work.

22 April 2011

doi:10.1016/j.jdermsci.2012.01.006