Borowczyk-Michałowska, Julia

Chrapusta, Anna

Drukała, Justyna

Konieczny, Piotr

Madeja, Zbigniew

Zimoląg, Eliza

Journal of Investigative Dermatology

English

Jagiellonian Univeristy Repository

LETTERS TO THE EDITORStage-Specific Embryonic Antigen-4 (SSEA-4)as a Distinguishing Marker between Eccrineand Apocrine Origin of Ducts of Sweat GlandsJournal of Investigative Dermatology (2017) 137, 2437e2440; doi:10.1016/j.jid.2017.07.003TO THE EDITORHuman skin has two major types ofsweat glands: eccrine and apocrine.They differ in function, structure, anddistribution over the surface of the body(Figure 1) (Sato et al., 1989). Althoughapocrine and eccrine secretory portionsof sweat glands are clearly morphologi-cally distinctive, their ducts are, to date,histologically and immunohistochemi-cally indistinguishable. The ability tomake this distinction is of a great signif-icance, especially in the case of anaxillary region, where both types ofsweat glands are distributed close toeachother. Theoriginof aparticular ductcan be identified only when its orifice isvisible; in other words, an eccrine ductopens directly into the epidermis,whereas an apocrine duct opens into theinfundibular portion of a hair follicle(Sato et al., 1989) (Figure 1a, 1b). In ourprevious study, we described stage-specific embryonic antigen-4 (SSEA-4)as a marker of ductal cells of eccrinesweat glands (Borowczyk-Michalowskaet al., 2017). Here, we investigatedwhether SSEA-4 is present in ductal cellsof apocrine sweat glands as well.Skin biopsy samples were obtainedfrom the axilla of nine healthy patientsundergoing plastic surgery: five femalepatients aged10, 31, 60, 74, and80yearsand four male patients aged 30, 31, 40,and 50 years. The written informedpatient consent andapprovalof the ethicscommittee were obtained under Polishlaw (no. KBET/72/B/2008). Skin sampleswere fixed and stained as describedby Borowczyk-Michalowska et al.(2017). The details of antibodies usedare described in the SupplementaryMaterials and Methods online.Abbreviations: CEA, carcinoembryonic antigen; K, keAccepted manuscript published online 21 July 2017ª 2017 The Authors. Published by Elsevier, Inc. on bThis is an open access article under the CC BY licensTo address the question of whetherSSEA-4 is also present in apocrinesweat glands, sections first were stainedfor carcinoembryonic antigen (CEA) tolocalize structures of both types ofsweat glands (see SupplementaryFigure S1a, S1b online). CEA expres-sion is solely restricted to sweat glandsin human skin, and it is present at theluminal surface of both eccrine andapocrine ductal cells, as well as in theeccrine secretory portion (Li et al.,2009; Saga, 2001). In the axillaryeccrine sweat glands, as in our previousstudy, SSEA-4 was found to be detectedin all cells along the full length of theduct, that is, in the intraepidermalacrosyringium and throughout thedermis to the coiled section leadingto the secretory portion (Figure 1a).Subsequently, the luminal cells co-expressed both types of embryonic an-tigens (Figure 2a). We identified ductsof apocrine sweat glands as structuresthat were positive for CEA and local-ized within hair follicles or within theclosest proximity of them (seeSupplementary Figure S1a). The ductalcells of apocrine origin had no expres-sion of SSEA-4, including in the upperpart where duct opens directly into thehair canal and in the dermal portionleading to the secretory coil (Figures 1aand 2a). To further confirm our findings,we analyzed the expression of SSEA-4concomitantly with several types ofkeratins. Although the presence of ker-atins is characteristic of all epithelialcells, they represent a complex family,with different proteins being expressedin specific types of cells. In eccrineducts, keratin (K) 16 was detected inboth layers of cells with luminal cellsratin; SSEA-4, stage-specific embryonic antigen-4ehalf of the Society for Investigative Dermatology.e (http://creativecommons.org/licenses/by/4.0/).being more predominantly stained,whereas ductal cells of apocrine sweatglands expressed K16 uniformly(Figure 2b, Figure S1d). Both eccrineand apocrine ducts, on the otherhand, showed a weak and variable re-action for K17, with only single cellsbeing strongly stained (Figure 2b,Supplementary Figure S1d). In contrastto apocrine ductal cells, which did notexpress SSEA-4, in eccrine ductsexpression of glycosphingolipid wasalways detected (Figure 2b). In rare in-stances, the single dots of SSEA-4 signalwere visible in luminal cells ofapocrine ducts; however, this signalwas very weak and still clearly distin-guishable from the one observed in theeccrine ducts (see SupplementaryFigure S1d). Additionally, our resultsshowed that no immunoreactivity forSSEA-4 was present in secretory cells ofboth eccrine and apocrine glands,although expression of K7, a marker ofsecretory coils, was clearly visible(Figure 2c). Thus, our findings providecompelling evidence supporting thenotion that SSEA-4 may be the previ-ously unreported marker that allows fordistinction between eccrine andapocrine ducts of sweat glands.In recent decades, several markershave been proposed to be characteristicof either eccrine or apocrine ductalcells. However, in practice, none ofthese markers have provided a reliableand reproducible way to make thisdistinction (Fuertes et al., 2013).Because CEA expression is detected inboth eccrine and apocrine sweat glands,it is not a useful discriminative marker.K16 is expressed in skin, not only inductal cells of both eccrine andapocrine origin, but also in basal cells ofepidermis (Langbein et al., 2008). K17,on the other hand, is present mainly inmyoepithelial cells of secretory coils butin hair follicles and sebaceous glands aswell (Moll and Moll, 1992; Scho¨n et al.,www.jidonline.org 2437Figure 1. Expression of SSEA-4 antigen in human apocrine and eccrine sweat glands. Immunohistochemical detection of SSEA-4 antigen expression within(a) the entire apocrine and (b) eccrine sweat glands. Inserts show magnified views of respective parts of glands. Schematic illustrations of two types ofsweat glands are included. (c) Major features of eccrine and apocrine sweat glands in humans. HF, hair follicle; SG, sebaceous gland; SSEA-4, stage-specificembryonic antigen-4. Scale bars ¼ 100 mm (low magnification) and 50 mm (high magnification). The ImageJ Stitching Plugin was used for image reconstruction(Preibisch et al., 2009).J Borowczyk-Michalowska et al.SSEA-4 Is Absent in the Apocrine Sweat Glands24381999). Thus, the presence of K16 andK17 in the multiple epithelial cells doesnot make them reliable markers forapocrine-eccrine distinction. Anotherantigen frequently mentioned to bespecific for the apocrine type of glands isJournal of Investigative Dermatology (2017), VolumGCDFP-15. However, as a componentof fluid secreted by cells, it is solelypresent in the secretory portion and notin the excretory ducts of apocrine sweatglands (see Supplementary Figure S1c).Moreover, according to the literature,e 137GCDFP-15 may also be detected ineccrine glands; thus, its application as adistinguishing marker is questionable(Viacava et al., 1998).The ability to distinguish the origin ofductal cells is of particular interest forFigure 2. Expression patterns of SSEA-4 and sweat gland markers in human axillary skin. Double immunofluorescence detection of sweat gland markersCEA, K7, K16, K17, and SSEA-4 antigen in sweat glands. (a) Staining for SSEA-4 (green) and CEA (red); (b) staining for SSEA-4 (green) and K16 or K17 (red);(c) staining for SSEA-4 (green) and K7 (red). Nuclei (blue) were counterstained with Hoechst 33258. Inserts show magnified views. Arrowheads indicate reactionfor K17. Scale bars ¼ 100 mm (low magnification), 50 mm (high magnification). AD, apocrine duct; AG, apocrine sweat gland; AS, apocrine secretoryportion; CEA, carcinoembryonic antigen; Der, dermis; ED, eccrine duct; EG, eccrine sweat gland; Epi, epidermis; ES, eccrine secretory portion; HF, hair follicle;K, keratin; SG, sebaceous gland; SSEA-4, stage-specific embryonic antigen-4.J Borowczyk-Michalowska et al.SSEA-4 Is Absent in the Apocrine Sweat Glandswww.jidonline.org 2439SUPPLEMENTARY MATERIALSupplementary material is linked to the onlineversion of the paper at www.jidonline.org, and athttp://dx.doi.org/10.1016/j.jid.2017.07.003.This work is licensed undera Creative Commons Attri-bution 4.0 International License. Toview a copy of this license, visithttp://creativecommons.org/licenses/by/4.0/C Tziotzios et al.MicroRNAs in Frontal Fibrosing Alopecia2440the development of tumor diagnosticmarkers, because currently the combi-nation of multiple antibodies is neces-sary to create a useful panel specific forapocrine and eccrine metastatic sweatgland carcinomas (Chintamani et al.,2003). In the light of these data and tothe best of our knowledge, SSEA-4 isthe first antigen enabling histologicaldistinction between ducts of eccrineand apocrine sweat glands. If theexpression profile of SSEA-4 is main-tained during the malignant trans-formation of cells derived from theducts of sweat glands, it can beassumed that it can be a very valuablediagnostic marker in the future.CONFLICT OF INTERESTThe authors state no conflict of interest.ACKNOWLEDGMENTSFaculty of Biochemistry, Biophysics and Biotech-nology of Jagiellonian University is a partner ofthe Leading National Research Centre (KNOW)supported by the Ministry of Science and HigherEducation.Julia Borowczyk-Michalowska1,4,Eliza Zimolag1,4, Piotr Konieczny2,Anna Chrapusta3, Zbigniew Madeja1and Justyna Drukala1,*1Department of Cell Biology, Faculty ofBiochemistry, Biophysics and Biotechnology,Journal of Investigative Dermatology (2017), VolumJagiellonian University, Krakow, Poland;2Department of General Biochemistry, Facultyof Biochemistry, Biophysics andBiotechnology, Jagiellonian University,Krakow, Poland; and 3Malopolska Burns andPlastic Surgery Centre, The Rydygier MemorialHospital, Krakow, Poland4These authors contributed equally to this work.*Corresponding author e-mail: justyna.drukala@uj.edu.plREFERENCESBorowczyk-Michalowska J, Zimolag E,Waligorska A, Dobrucki J, Madeja Z, Drukala J.Stage-specific embryonic antigen-4 as a novelmarker of ductal cells of human eccrine sweatglands. Br J Dermatol 2017;176:1541e8.Chintamani, Sharma R, Badran R, Singhal V,Saxena S, Bansal A. Metastatic sweat glandadenocarcinoma: A clinico-pathologicaldilemma. World J Surg Oncol 2003;1:13.Fuertes L, Santonja C, Kutzner H, Requena L. Immu-nohistochemistry in dermatopathology: a reviewofthemost commonly used antibodies (part II). ActasDermosifiliogr 2013;104:181e203.Langbein L, Cribier B, Schirmacher P, Praetzel-Wunder S, Peltre B, Schweizer J. New conceptson the histogenesis of eccrine neoplasia fromkeratin expression in the normal eccrine gland,syringoma and poroma. Br J Dermatol 2008;159:633e45.e 137Li H-H, Zhou G, Fu X-B, Zhang L. Antigenexpression of human eccrine sweat glands.J Cutan Pathol 2009;36:318e24.Moll I, Moll R. Changes of expression of inter-mediate filament proteins during ontogenesis ofeccrine sweat glands. J Invest Dermatol1992;98:777e85.Preibisch S, Saalfeld S, Tomancak P. Globallyoptimal stitching of tiled 3D microscopic imageacquisitions. Bioinformatics 2009;25:1463e5.Saga K. Histochemical and immunohistochemicalmarkers for human eccrine and apocrine sweatglands: an aid for histopathologic differentia-tion of sweat gland tumors. J Investig DermatolSymp Proc 2001;6:49e53.Sato K, KangWH, Saga K, Sato KT. Biology of sweatglands and their disorders. I. Normal sweat glandfunction. J Am Acad Dermatol 1989;20:537e63.Scho¨n M, Benwood J, O’Connell-Willstaedt T,Rheinwald JG. Human sweat gland myoepithe-lial cells express a unique set of cytokeratins andreveal the potential for alternative epithelial andmesenchymal differentiation states in culture.J Cell Sci 1999;112(Pt. 12):1925e36.Viacava P, Naccarato AG, BevilacquaG. Spectrum ofGCDFP-15 expression in human fetal and adultnormal tissues. Virchows Arch 1998;432:255e60.

Stage-specific embryonic antigen-4 (SSEA-4) as a distinguishing marker between eccrine and apocrine origin of ducts of sweat glands

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Stage-specific embryonic antigen-4 (SSEA-4) as a distinguishing marker between eccrine and apocrine origin of ducts of sweat glands

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