Isolation of T cells from mouse and human skin

Macrophage tropic HIV-1 is predominant during the initial viremia after person to person transmission of HIV-1 (Zhu, T., H. Mo, N. Wang, D.S. Nam, Y. Cao, R.A. Koup, and D.D. Ho. 1993. Science. 261:1179–1181.), and this selection may occur during virus entry and carriage to the lymphoid tissue. Human skin explants were used to model HIV-1 selection that may occur at the skin or mucosal surface. Macrophage tropic, but not T cell line tropic strains of HIV-1 applied to the abraded epidermis were recovered from the cells emigrating from the skin explants. Dermis and epidermis were separated by dispase digestion after virus exposure to determine the site of viral selection within the skin. Uptake and transmission to T cells of all HIV-1 isolates was found with the dermal emigrant cells, but only macrophage tropic virus was transferred by emigrants from the epidermis exposed to HIV-1, indicating selection only within the epidermis. CD3+, CD4+ T cells were found in both the dermal and epidermal emigrant cells. After cell sorting to exclude contaminating T cells, macrophage tropic HIV-1 was found in both the dermal emigrant dendritic cells and in dendritic cells sorted from the epidermal emigrants. These observations suggest that selective infection of the immature epidermal dendritic cells represents the cellular mechanism that limits the initial viremia to HIV-1 that can use the CCR5 coreceptor.

Download Full-text

Regulatory T Cells Inhibit CD8+ T-Cell Tissue Invasion in Human Skin Graft-Versus-Host Reactions

Transplantation Journal ◽

10.1097/tp.0b013e31826205d6 ◽

2012 ◽

Vol 94 (5) ◽

pp. 456-464 ◽

Cited By ~ 15

Author(s):

Emily Mavin ◽

Shaheda S. Ahmed ◽

Graeme O’Boyle ◽

Brie Turner ◽

Stephen Douglass ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Regulatory T Cells ◽

Human Skin ◽

Skin Graft ◽

Cd8 T Cell ◽

Graft Versus Host ◽

Tissue Invasion ◽

Cell Tissue

Download Full-text

IL-23 induced in keratinocytes by endogenous TLR4 ligands polarizes dendritic cells to drive IL-22 responses to skin immunization

Journal of Experimental Medicine ◽

10.1084/jem.20150376 ◽

2016 ◽

Vol 213 (10) ◽

pp. 2147-2166 ◽

Cited By ~ 32

Author(s):

Juhan Yoon ◽

Juan Manuel Leyva-Castillo ◽

Guoxing Wang ◽

Claire Galand ◽

Michiko K. Oyoshi ◽

...

Keyword(s):

T Cells ◽

Human Skin ◽

Cd4 T Cells ◽

Mouse Skin ◽

Skin Inflammation ◽

Serum Levels ◽

Skin Lesions ◽

Tape Stripping ◽

Keratinocyte Proliferation ◽

Th22 Cells

Atopic dermatitis (AD) is a Th2-dominated inflammatory skin disease characterized by epidermal thickening. Serum levels of IL-22, a cytokine known to induce keratinocyte proliferation, are elevated in AD, and Th22 cells infiltrate AD skin lesions. We show that application of antigen to mouse skin subjected to tape stripping, a surrogate for scratching, induces an IL-22 response that drives epidermal hyperplasia and keratinocyte proliferation in a mouse model of skin inflammation that shares many features of AD. DC-derived IL-23 is known to act on CD4+ T cells to induce IL-22 production. However, the mechanisms that drive IL-23 production by skin DCs in response to cutaneous sensitization are not well understood. We demonstrate that IL-23 released by keratinocytes in response to endogenous TLR4 ligands causes skin DCs, which selectively express IL-23R, to up-regulate their endogenous IL-23 production and drive an IL-22 response in naive CD4+ T cells that mediates epidermal thickening. We also show that IL-23 is released in human skin after scratching and polarizes human skin DCs to drive an IL-22 response, supporting the utility of IL-23 and IL-22 blockade in AD.

Download Full-text

Bispecific T-Cell Engager -Armored Chimeric Antigen Receptor T Cells Overcome Antigen Escape From EGFRvIII-Targeted Therapy For Glioblastoma

Neurosurgery ◽

10.1093/neuros/nyz310_154 ◽

2019 ◽

Vol 66 (Supplement_1) ◽

Author(s):

Bryan D Choi ◽

Xiaoling Yu ◽

Ana P Castano ◽

Amanda A Bouffard ◽

Andrea Schmidts ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Human Skin ◽

Immune Therapy ◽

Biologically Active ◽

Target Antigen ◽

Wild Type ◽

Car T Cells ◽

Car T ◽

The Brain

Abstract INTRODUCTION Immune therapy with T cells engineered to express chimeric antigen receptors (CARs) represents a promising therapy for patients with glioblastoma (GBM). However, clinical responses have been limited due to heterogeneous target antigen expression and outgrowth of tumors lacking the antigen targeted by CAR T cells directed against a single target. In clinical studies with CART-EGFRvIII, EGFRvIII-targeted T cells successfully localized to the brain tumor microenvironment, but ultimately failed to prevent disease progression with post-treatment specimens demonstrating high levels of wild-type EGFR despite reduced expression of EGFRvIII. METHODS We developed a novel bicistronic CAR construct engineered for local delivery of bispecific T-cell engagers (BiTEs) that target residual tumor. Specifically, EGFRvIII-targeted CAR T cells were engineered to secrete BiTEs against wild-type EGFR, which is frequently amplified and overexpressed in GBM. RESULTS Human T cells were efficiently transduced with the dual CART.BiTE transgene. These modified cells secreted biologically active EGFR-specific BiTEs that not only redirected CAR T cells but also recruited and activated untransduced bystander T cells against wild-type EGFR. Recapitulating clinical data, EGFRvIII CAR T cells were unable to completely treat tumors with heterogenous EGFRvIII expression, leading to outgrowth of EGFRvIII-negative, EGFR-positive GBM. Conversely, CART.BiTE cells cured mice even in the setting of antigen-loss, against heterogeneous and well-established intracerebral tumors in mice. Unlike CAR T cells directly targeting EGFR, which caused toxicity in human skin grafts in vivo, secreted BiTE-EGFR was both locally effective and did not result in toxicity against grafted human skin. CONCLUSION This is the first instance in which CARs and BiTEs have been combined into a single platform of immune therapy. Our results demonstrate that CARs and BiTEs can be combined strategically to mitigate antigen heterogeneity in GBM and also provide a unique T-cell-based delivery method for BiTEs to tumors in the brain.

Download Full-text