Ann Dermatol.  2017 Apr;29(2):156-166. 10.5021/ad.2017.29.2.156.

The Contributory Roles of Th17 Lymphocyte and Cytotoxic T Lymphocyte at the Hair Bulge Region as Well as the Hair Bulb Area in the Chronic Alopecia Areata Patients

• Hong JW
• Lee CY
• Ha SM
• Choi SH
• Kim TH
• Song KH
• Kim KH

• KMID: 2394838
• DOI: http://doi.org/10.5021/ad.2017.29.2.156

Abstract

BACKGROUND
Alopecia areata (AA) is a T cell-mediated autoimmune disease that targets hair follicles and interrupts hair regrowth. The microenvironment of the effector T cells and their related cytokines may affect immunopathogenesis around the hair bulb/bulge.
OBJECTIVE
To determine the contributory roles of the effector T cell subsets and related cytokines to the pathogenesis of AA.
METHODS
We investigated the correlation between histopathological grades and four clinical prognostic factors in 331 patients with AA, and analyzed the topography of T cell infiltrates and related cytokines around the hair bulb/bulge according to histopathological grades through immunohistochemical and double immunofluorescence studies on a subset of AA specimens.
RESULTS
First, the groups with more severe histopathological grades were associated with earlier onset, longer duration, more hair loss, as well as poorer therapeutic outcomes. Second, the pattern of CD4 and CD8 expression around the hair bulb/bulge varied by histopathological grade, with staining density decreasing in the following order: type 1>type 2>type 3. In addition, interferon-γ and transforming growth factor-β1 expression appeared denser in the peribulbar area. Interestingly, the denser CCR6⁺ cells (Th17 cells) showed more infiltration than CCR5⁺ cells (Th1 cells) around the hair bulb/bulge as histopathological grade worsened.
CONCLUSION
The insidious destruction of bulge stem cells and hair bulb matrix stem cells results in more severe hair loss in patients with chronic AA, which is mediated by Th17 lymphocyte and cytotoxic T lymphocyte infiltration. Furthermore, Th17 lymphocytes may play an even more important role than cytotoxic T cells in the development of AA.

Keyword

Chronic alopecia areata; Cytotoxic T lymphocyte; Th17 cells

MeSH Terms

Alopecia Areata*
Alopecia*
Autoimmune Diseases
Cytokines
Fluorescent Antibody Technique
Hair Follicle
Hair*
Humans
Lymphocytes*
Stem Cells
T-Lymphocyte Subsets
T-Lymphocytes
Th17 Cells
Cytokines

Figure

• 

  Fig. 1 Immunnohistochemistries for CD4+ and CD8+ lymphocytes. CD4 positive cells infiltrated mostly in peribulbar area. CD8+ cells infiltrated mostly in intrabulbar area. The severer histopathologic gradings were associated with the denser infiltrations of mononuclear cell (A~C: CD4, ×100), (D~F, CD8, ×100).

• 

  Fig. 2 CD4 and CD8 immunohistochemistries for types 1, 2, and 3 hair follicles in alopecia areata (AA) patients: (A) and (D), type 3 AA hair follicles, (B) and (E); type 2 AA hair follicles, (C) and (F); type 1 AA hair follicles. At hair bulb (Bb) areas, CD4+ T lymphocytes infiltrated perifollicularly and CD8+ T lymphocytes infiltrated intrafollicularly, Interestingly, the denser CD4+ and CD8+ T lymphocytes infiltrated around hair bulge (Bg) regions as the histopathologic gradings were worsened.

• 

  Fig. 3 Expression of CCR6 and CCL20 within type 1 hair bulbs (B~D), type 2 hair bulbs (F~H), and type 3 hair bulbs (J~L). A representative finding of double immunofluoroscence showed immunoreactive cells for CCR6 (B, F, J; green) and CCL20 (C, G, K; red) withco-localization study of CCR6 and CCL20 (D, H, L). Same samples for H&E stain were also shown from type 1 to type 3 (A, E ,I). The denser CCR6 and CCL20 were expressed in parallel with the severer histopathologic gradings (H&E: A, E, I, ×100; double immunofluorescence: B~D, F~H, J~L, ×100).

• 

  Fig. 4 Expression of CCR6 and CCR5 within type 1 hair bulbs (B~D), type 2 hair bulbs (F~H), and type 3 hair bulbs (J~L). A representative finding of double immunofluorescences immunoreactive cells for CCR6 (B, F, J; green) and CCR5 (C, G, K; red) with colocalization study of CCR6 and CCR5 (D, H, L). H&E stained sectionswere also shown from type 1 to type 3 (A, E, I). The denser CCR6 and CCR5 were expressed in parallel with the severer histopathologic gradings (H&E: A, E, I, ×100; double immunofluorescence, B~D, F~H, J~L, ×100).

• 

  Fig. 5 Expression of CCR6 and CCL20 within type 1 hair bulges (A~C), type 2 hair bulges (D~F), and type 3 hair bulges (G~I).

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  Fig. 6 Immunnohistochemistry for interferon (IFN)-γ. IFN-γ wasexpressed in the inflammatory cells around the hair bulb, and the severer histopathologic grading was associated with the denser immunoreactive cells for IFN-γ (A~C, ×100; D~F, ×200).

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  Fig. 7 Immunnohistochemistry for transforming growth factor (TGF)-β1. Expression of TGF-β1 was detected in the outer root sheath keratinocyte, regressing epithelial strand, dermal sheath and dermal sheath cells, and its expression was correlated with histopathologic grading (A~C, ×100; D~F, ×200).

• 

  Fig. 8 Immunnohistochemistry for transforming growth factor (TGF)-β2. TGF-β2 was expressed in the dermal papilla area, and less expression was correlated with the severer histopathologic gradings (A~C, ×100; D~F, ×200).

Reference

1. Christoph T, Müller-Röver S, Audring H, Tobin DJ, Hermes B, Cotsarelis G, et al. The human hair follicle immune system: cellular composition and immune privilege. Br J Dermatol. 2000; 142:862–873.

2. Meyer KC, Klatte JE, Dinh HV, Harries MJ, Reithmayer K, Meyer W, et al. Evidence that the bulge region is a site of relative immune privilege in human hair follicles. Br J Dermatol. 2008; 159:1077–1085.

3. McElwee KJ, Freyschmidt-Paul P, Hoffmann R, Kissling S, Hummel S, Vitacolonna M, et al. Transfer of CD8(+) cells induces localized hair loss whereas CD4(+)/CD25(−) cells promote systemic alopecia areata and CD4(+)/CD25(+) cells blockade disease onset in the C3H/HeJ mouse model. J Invest Dermatol. 2005; 124:947–957.

4. Tanemura A, Oiso N, Nakano M, Itoi S, Kawada A, Katayama I. Alopecia areata: infiltration of Th17 cells in the dermis, particularly around hair follicles. Dermatology. 2013; 226:333–336.

5. Lew BL, Cho HR, Haw S, Kim HJ, Chung JH, Sim WY. Association between IL17A/IL17RA gene polymorphisms and susceptibility to alopecia areata in the Korean population. Ann Dermatol. 2012; 24:61–65.

6. Cotsarelis G, Millar SE. Towards a molecular understanding of hair loss and its treatment. Trends Mol Med. 2001; 7:293–301.

7. Mobini N, Tam S, Kamino H. Possible role of the bulge region in the pathogenesis of inflammatory scarring alopecia: lichen planopilaris as the prototype. J Cutan Pathol. 2005; 32:675–679.

8. Whiting DA. Histopathologic features of alopecia areata: a new look. Arch Dermatol. 2003; 139:1555–1559.

9. Al-Refu K. Stem cells and alopecia: a review of pathogenesis. Br J Dermatol. 2012; 167:479–484.

10. Yoshida R, Tanaka K, Amagai M, Ohyama M. Involvement of the bulge region with decreased expression of hair follicle stem cell markers in senile female cases of alopecia areata. J Eur Acad Dermatol Venereol. 2011; 25:1346–1350.

11. Acosta-Rodriguez EV, Rivino L, Geginat J, Jarrossay D, Gattorno M, Lanzavecchia A, et al. Surface phenotype and antigenic specificity of human interleukin 17-producing T helper memory cells. Nat Immunol. 2007; 4:639–646.

12. Zambrano-Zaragoza JF, Romo-Martínez EJ, Durán-AvelarMde J, García-Magallanes N, Vibanco-Pérez N. Th17 cells in autoimmune and infectious diseases. Int J Inflam. 2014; 2014:651503.

13. Oiso N, Kawada A. Renbök phenomenon in a patient with alopecia areata universalis and psoriasis. J Dermatol. 2012; 39:288–289.

14. König A, Happle R, Hoffmann R. IFN-gamma-induced HLA-DR but not ICAM-1 expression on cultured dermal papilla cells is downregulated by TNF-alpha. Arch Dermatol Res. 1997; 289:466–470.

15. Gilhar A, Landau M, Assy B, Shalaginov R, Serafimovich S, Kalish RS. Mediation of alopecia areata by cooperation between CD4+ and CD8+ T lymphocytes: transfer to human scalp explants on Prkdc(scid) mice. Arch Dermatol. 2002; 138:916–922.

16. Gilhar A, Kam Y, Assy B, Kalish RS. Alopecia areata induced in C3H/HeJ mice by interferon-gamma: evidence for loss of immune privilege. J Invest Dermatol. 2005; 124:288–289.

17. Paus R, Ito N, Takigawa M, Ito T. The hair follicle and immune privilege. J Investig Dermatol Symp Proc. 2003; 8:188–194.

18. Teraki Y, Imanishi K, Shiohara T. Cytokines in alopecia areata: contrasting cytokine profiles in localized form and extensive form (alopecia universalis). Acta Derm Venereol. 1996; 76:421–423.

19. Arca E, Muşabak U, Akar A, Erbil AH, Taştan HB. Interferon-gamma in alopecia areata. Eur J Dermatol. 2004; 14:33–36.

20. Zhu J, Paul WE. CD4 T cells: fates, functions, and faults. Blood. 2008; 112:1557–1569.

21. Paus R, Christoph T, Müller-Röver S. Immunology of the hair follicle: a short journey into terra incognita. J Investig Dermatol Symp Proc. 1999; 4:226–234.

22. Paus R, Foitzik K, Welker P, Bulfone-Paus S, Eichmüller S. Transforming growth factor-beta receptor type I and type II expression during murine hair follicle development and cycling. J Invest Dermatol. 1997; 109:518–526.

23. Welker P, Foitzik K, Bulfone-Paus S, Henz BM, Paus R. Hair cycle-dependent changes in the gene expression and protein content of transforming factor beta 1 and beta 3 in murine skin. Arch Dermatol Res. 1997; 289:554–557.

24. Soma T, Tsuji Y, Hibino T. Involvement of transforming growth factor-beta2 in catagen induction during the human hair cycle. J Invest Dermatol. 2002; 118:993–997.

25. Inoue K, Aoi N, Yamauchi Y, Sato T, Suga H, Eto H, et al. TGF-beta is specifically expressed in human dermal papilla cells and modulates hair folliculogenesis. J Cell Mol Med. 2009; 13:4643–4656.