Human skin cells support thymus-independent T cell development
Rachael A. Clark, Kei-ichi Yamanaka, Mei Bai, Rebecca Dowgiert, Thomas S. Kupper
Rachael A. Clark, Kei-ichi Yamanaka, Mei Bai, Rebecca Dowgiert, Thomas S. Kupper
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Research Article Immunology

Human skin cells support thymus-independent T cell development

Abstract

Thymic tissue has previously been considered a requirement for the generation of a functional and diverse population of human T cells. We report that fibroblasts and keratinocytes from human skin arrayed on a synthetic 3-dimensional matrix support the development of functional human T cells from hematopoietic precursor cells in the absence of thymic tissue. Newly generated T cells contained T cell receptor excision circles, possessed a diverse T cell repertoire, and were functionally mature and tolerant to self MHC, indicating successful completion of positive and negative selection. Skin cell cultures expressed the AIRE, Foxn1, and Hoxa3 transcription factors and a panel of autoantigens. Skin and bone marrow biopsies can thus be used to generate de novo functional and diverse T cell populations for potential therapeutic use in immunosuppressed patients.

Authors

Rachael A. Clark, Kei-ichi Yamanaka, Mei Bai, Rebecca Dowgiert, Thomas S. Kupper

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T cells produced in skin cell cultures are mature and functional. (A and...
T cells produced in skin cell cultures are mature and functional. (A and B) Proliferation of T cells in response to (A) control medium and (B) phytohemagglutinin (PHA). (C–F) Expression of CD69 activation antigen in response to (C) control medium and (D–F) concanavalin A (Con A) by (C and D) total T cells and (E) CD4+ and (F) CD8+ subsets. (G and H) Production of TNF-α by CD3+ cells in response to (G) control medium and (H) concanavalin A treatment. (I and J) Production of (I) IFN-γ and (J) IL-2 in response to concanavalin A treatment. Treatment with control medium is not shown for IFN-γ and IL-2 samples but was identical to TNF-α control shown in G. The percentage of positive cells is shown. (K) Proliferation of T cells generated in skin cell cultures (T cells) in response to allogeneic PBMCs. Proliferation was assayed by BrdU incorporation on day 6 of the MLR. BrdU incorporation was detected via flow cytometry with gating on CD3+ T cells. PBMCs were derived from allogeneic, unrelated donors [PBMCs (A), donor A; PBMCs (B), donor B]. Error bars represent the SD of 3 measurements.