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. Author manuscript; available in PMC: 2015 Sep 18.
Published in final edited form as: Sci Transl Med. 2015 Mar 18;7(279):279ra39. doi: 10.1126/scitranslmed.3010302

Figure 6. T cell CD103 induction is enhanced by keratinocyte contact, depends on TGFβ and is independent of T cell keratinocyte adhesive interactions.

Figure 6

(A,B) In human engrafted mice, T cells that migrating into the epidermis had a higher rate of CD103 up-regulation than dermal T cells. (A) Representative immunostains of human foreskin skin grafts 3 weeks after IV injection of CD103− PBMC are shown. CD103+ T cells were primarily localized to the epidermal compartment. The dermal-epidemal junction is highlighted by a white line. (B) The mean and SEM of 8 separate experiments are shown (test: T-test). (C) Keratinocyte contact induced up regulation of CD103 in CD4+ T cells from human blood stimulated with αCD3/αCD2/αCD28 beads. CD4+ T cells were isolated from human peripheral blood by magnetic bead separation and then co-cultured in direct contact (direct) with confluent monolayers of human keratinocytes (ker) or fibroblasts (fib) or in transwells separated from keratinocyte or fibroblasts monolayers (indirect) for one week in the presence of αCD3/αCD2/αCD28 beads. CD103 expression was then assessed by immunostaining and flow cytometry anlysis. The mean and SEM of four separate experiments are shown (test: ANOVA). (D). Blocking of adhesive interactions with keratinocytes had no effect on CD103 upregulation but neutralization of TGFβ did decrease CD103 induction. CD4+ T cells were cultured on confluent human keratinocyte monolayers for one week in the presence of αCD3/αCD2/αCD28 beads and the indicated control and function blocking antibodies. The mean and SEM of four experiments are shown (test: ANOVA). (E,F) TGFβ neutralizing antibodies decreased and exogenous TGFβ increased CD103 induction. T cells were cultured in transwells across from keratinocyte monolayers (E) or indirect contact with keratinocyte monolayers (F) for one week in the presence of αCD3/αCD2/αCD28 beads and the indicated neutralizing TGFβ antibodies (αTGFβ) or recombinant human TGFβ (hTGFβ). the mean and SEM of 2 separate experiments are shown (test: ANOVA). (G) Function blocking E-cadherin antibodies inhibited T cell adhesion to keratinocyte monolayers. CD4+ T cells were cultured on confluent human keratinocyte monolayers for one week in the presence of αCD3/αCD2/αCD28 beads in the presence of isotype control or E-cadherin function blocking antibodies. Nonadherent T cells were removed by rinsing and adherent T cells were then eluted from the wells by treatment with trypsin/EDTA. The mean and SEM of the number of adherent cells per well from three separate experiments are shown (test:T-test).