Allergy Asthma Immunol Res.  2019 Sep;11(5):677-690. 10.4168/aair.2019.11.5.677.

Activated Leukocyte Cell Adhesion Molecule Modulates Th2 Immune Response in Atopic Dermatitis

• Oh MS
• Hong JY
• Kim MN
• Kwak EJ
• Kim SY
• Kim EG
• Lee KE
• Kim YS
• Jee HM
• Kim SH
• Sol IS
• Park CO
• Kim KW
• Sohn MH

• KMID: 2452758
• DOI: http://doi.org/10.4168/aair.2019.11.5.677

Abstract

PURPOSE
Activated leukocyte cell adhesion molecule (ALCAM), a member of the immunoglobulin superfamily, is highly expressed on dendritic cells. ALCAM and its receptor CD6 are co-stimulatory molecules in the immunological synapse; their interaction is required for T cell activation. While atopic dermatitis (AD) is recognized as a T helper 2 (Th2)-mediated allergic disease, the role of ALCAM in its pathogenesis is unclear.
METHODS
ALCAM levels were measured in the serum of AD patients and AD-induced murine model by ovalbumin treatment. We next investigated transepidermal water loss, clinical score, Th2-immune responses, skin barrier gene expression and T-cell activation using wild-type (WT) and ALCAM deficiency mice. An oxazolone-induced AD-like model was also established and analyzed using WT- and ALCAM-deficient mice.
RESULTS
We found that serum ALCAM levels were elevated in pediatric AD patients as well as WT AD mice, whereas Th2-type cytokine production and AD symptoms were suppressed in ALCAM-deficient mice. In addition, CD4+ effector T-cell counts in murine skin and skin-draining lymph nodes were lower in ALCAM-deficient mice than in their WT counterparts. ALCAM deficiency was also linked to higher expression of skin barrier genes and number of lamellar bodies.
CONCLUSIONS
These findings indicate that ALCAM may contribute to AD pathogenesis by meditating a Th2-dominant immune response and disrupting the barrier function of the skin.

Keyword

ALCAM; CD166; atopic dermatitis; type 2 helper T cells; skin barrier

MeSH Terms

Activated-Leukocyte Cell Adhesion Molecule*
Animals
Dendritic Cells
Dermatitis, Atopic*
Gene Expression
Humans
Immunoglobulins
Immunological Synapses
Lymph Nodes
Mice
Ovalbumin
Skin
T-Lymphocytes
Water
Activated-Leukocyte Cell Adhesion Molecule
Immunoglobulins
Ovalbumin
Water

Figure

• 

  Fig. 1 ALCAM expression in pediatric AD patients and OVA-induced AD mice. (A-C) ALCAM level was measured by ELISA in serum samples from healthy controls (n = 44) and AD patients (n = 114). Pirate plots show the distribution of ALCAM levels for each group; horizontal lines represent means and boxes represent 95% confidence intervals. (A) ALCAM level in human serum. (B) Increase in serum ALCAM level as a function of AD severity increases. (C) Correlation between serum ALCAM level and AD SCORAD index. (D) Serum ALCAM abundance was assessed by ELISA. The data represent mean ± standard error of the mean. ALCAM, activated leukocyte cell adhesion molecule; AD, atopic dermatitis; OVA, ovalbumin; PBS, phosphate-buffered saline; SCORAD, SCORing atopic dermatitis; ELISA, enzyme-linked immunosorbent assay. *P < 0.05; †P < 0.001 by t-test (n = 5–10 mice/group).

• 

  Fig. 2 ALCAM attenuates skin barrier disruption induced by epicutaneous OVA sensitization. (A) Time course of clinical score and (B) TEWL. (C) Electron micrographs of osmium tetroxide-postfixed skins show LBs in SG (×50,000, Scale bar = 0.5 μm; Insert C). (Left) Normal shaped LBs are present in sham mouse epidermis. (Middle) WT/OVA mouse epidermis shows severely abnormal LBs, (Right) Whereas LBs in ALCAM−/− mice display less abnormality (×120,000, Scale bar = 0.2 μm). (D) Bar graph represents the number of LBs in SG. (E) The mRNA expression of skin barrier genes (filaggrin, loricrin and involucrin). The data represent mean ± standard error of the mean. ALCAM, activated leukocyte cell adhesion molecule; TEWL, transepidermal water loss; OVA, ovalbumin; PBS, phosphate-buffered saline; NS, not significant; WT, wild-type; LB, lamellar body; mRNA, messenger RNA; SG, stratum granulosum. *P < 0.05 (WT/PBS vs. WT/OVA); †P < 0.05 (ALCAM−/−/PBS vs. ALCAM−/−/OVA); ‡P < 0.05 (WT/OVA vs. ALCAM−/−/OVA); §P < 0.05; ∥P < 0.01; ¶P < 0.001 (n = 5–7 mice/group).

• 

  Fig. 3 ALCAM deficiency attenuates T helper-dominant skin inflammation induced by epicutaneous OVA sensitization. (A) Mouse skin biopsy specimen stained with hematoxylin and eosin (×200, Scale bar = 50 μm) and scored according to histological features. (B) Serum levels of total IgE measured by enzyme-linked immunosorbent assay. (C-H) Skin mRNA levels were assessed by quantitative real-time polymerase chain reaction; (C) IL-4, (D) IL-5, (E) IL-13, (F) IFN-γ, (G) IL-17A and (H) IL-22. The data represent mean ± standard error of the mean. ALCAM, activated leukocyte cell adhesion molecule; NS, not significant; OVA, ovalbumin; PBS, phosphate-buffered saline; WT, wild-type; IgE, immunoglobulin E; IL, interleukin; IFN, interferon; mRNA, messenger RNA. *P < 0.05; †P < 0.01; ‡P < 0.001 (n = 5–7 mice/group).

• 

  Fig. 4 ALCAM deficiency suppresses CD4+ T cell activation. Cells from the (A and B) skin and (C and D) skin draining -LN(dLN)s were harvested and stained for CD3, CD4, CD44 and CD62L. Cells were gated on CD3+CD4+ cells and then on CD44 and CD62L. Naïve cells are identified as CD44loCD62Lhi and effector cells as CD4hiCD62Llo. Representative dot plots are presented. Graphs represent percentage of (B) CD3+CD4+CD44loCD62Lhi in skin and (D) CD3+CD4+CD4hiCD62Llo in dLNs. The data represent mean ± standard error of the mean. ALCAM, activated leukocyte cell adhesion molecule; WT, wild-type; NS, not significant; OVA, ovalbumin; dLN, skin-draining lymph node. *P < 0.05; †P < 0.01; ‡P < 0.001(n = 5–7 mice/group).

• 

  Fig. 5 ALCAM deficiency attenuates murine AD-like skin lesion induced by repeated OXA application. (A-C) AD severity was assessed by (A) clinical score, (B) TEWL and (C) ear thickness. (D) Mouse ear skin biopsy specimen stained with hematoxylin and eosin (×100, bar = 200 μm) and (E) scored according to histological features. (F) Total IgE level in serum, as determined by enzyme-linked immunosorbent assay. The mRNA expression of (G) IL-4 and IL-13 in skin lesions. Data represent mean ± standard error of the mean. ALCAM, activated leukocyte cell adhesion molecule; OXA, oxazolone; AD, atopic dermatitis; mRNA, messenger RNA; NS, not significant; WT, wild-type; TEWL, transepidermal water loss; IgE, immunoglobulin E; IL, interleukin, PBS, phosphate-buffered saline. *P < 0.05 (WT/PBS vs. WT/OXA); †P < 0.05 (ALCAM−/−/PBS vs. ALCAM−/−/OXA); ‡P < 0.05 (WT/OXA vs. ALCAM−/−/OXA); §P < 0.05; ∥P < 0.01; ¶P < 0.001 (n = 4–6 mice/group).

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