A novel method for the isolation of skin resident T cells from normal and diseased human skin.
T cells resident in normal skin likely conduct immunosurveillance and are implicated in the development of inflammatory disorders such as psoriasis. This population of cells is difficult to study because existing techniques allow isolation of only few cells. We report here a novel method of isolating T cells from both normal and diseased human skin. Explants of skin cultured on three-dimensional matrices led to the outgrowth of dermal fibroblasts that elaborated T cell chemoattractant factors. These factors led to the migration of skin resident T cells out of skin explants where they could be collected and studied. Skin resident T cells isolated from explant cultures were CD45RO(+) memory T cells and expressed high levels of cutaneous lymphocyte antigen (CLA) and chemokine receptor (CCR)4. Inclusion of IL-2 and IL-15 in explant cultures produced up to a 10-fold expansion of skin-resident T cells, while maintaining the CLA(+)CCR4(+) skin-homing phenotype as well as a diverse T cell repertoire. This method also allowed efficient isolation of malignant T cells from the skin lesions of cutaneous T cell lymphoma and the isolation of tumor-infiltrating lymphocytes from primary squamous cell carcinomas and melanoma metastases.
Department of Dermatology, Brigham and Women's Hospital and the Harvard Skin Disease Research Center, 77 Avenue Louis Pasteur, Boston, MA 02115, USA.
Rachael A
RA
Clark
Benjamin F
BF
Chong
Nina
N
Mirchandani
Kei-Ichi
K
Yamanaka
George F
GF
Murphy
Rebecca K
RK
Dowgiert
Thomas S
TS
Kupper
D000945:Antigens, Differentiation, T-Lymphocyte; D000951:Antigens, Neoplasm; D002294:Carcinoma, Squamous Cell / Q000473:pathology; D002465:Cell Movement; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D018925:Chemokines / Q000096:biosynthesis; D005347:Fibroblasts / Q000502:physiology; D006801:Humans; D019409:Interleukin-15 / Q000494:pharmacology; D007376:Interleukin-2 / Q000494:pharmacology; D016246:Lymphocytes, Tumor-Infiltrating / Q000473:pathology; D016410:Lymphoma, T-Cell, Cutaneous / Q000473:pathology; D008545:Melanoma / Q000473:pathology / Q000556:secondary; D008562:Membrane Glycoproteins / Q000032:analysis; D054398:Receptors, CCR4; D019707:Receptors, Chemokine / Q000032:analysis; D012867:Skin / Q000166:cytology* / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology*
D000945:Antigens, Differentiation, T-Lymphocyte; D000951:Antigens, Neoplasm; C515354:CCR4 protein, human; C063419:CTAGE1 protein, human; D018925:Chemokines; D019409:Interleukin-15; D007376:Interleukin-2; D008562:Membrane Glycoproteins; D054398:Receptors, CCR4; D019707:Receptors, Chemokine