In vitro propagation and dynamics of T cells from skin biopsies by methods using interleukins-2 and -4 or anti-CD3/CD28 antibody-coated microbeads

Hideo Hashizume; Anker Hansen; Lars K Poulsen; Allan Randrup Thomsen; Masahiro Takigawa; Kristian Thestrup-Pedersen

doi:10.2340/00015555-0927

In vitro propagation and dynamics of T cells from skin biopsies by methods using interleukins-2 and -4 or anti-CD3/CD28 antibody-coated microbeads

Hideo Hashizume, Anker Hansen, Lars K Poulsen, Allan Randrup Thomsen, Masahiro Takigawa, Kristian Thestrup-Pedersen

Department of Immunology and Microbiology

13 Citations (Scopus)

Abstract

In order to explore the mechanisms of inflammatory skin disorders, we established two methods of expanding skin-derived lymphocytes, one using high levels of interleukin (IL)-2 and IL-4 (method A) and the other using low levels of cytokines and anti-CD3/CD28 microbeads (method B). Both methods provide advantages for functional studies. With either of these two, we could obtain more than 10⁷ cells/ from a 3 mm skin biopsy in 21 days from 23 out of 26 biopsies of various skin diseases. The relevance of these cells was confirmed by shifted T-cell receptor β chain variable region (TCR-Vβ) repertoire and antigen-dependent proliferation in antigen-driven skin disorders. The propagation of skin-resident lymphocytes, seen especially in method A, seems to be mediated by a functional defect of regulatory T cells residing in skin sequentially expanding under the conditions of our methods.

Original language	English
Journal	Acta Dermato Venereologica
Volume	90
Issue number	5
Pages (from-to)	468-73
Number of pages	6
ISSN	0001-5555
DOIs	https://doi.org/10.2340/00015555-0927
Publication status	Published - 1 Sept 2010

Keywords

Adult
Aged
Antibodies
Antigens, CD28
Antigens, CD3
Biopsy
Cell Culture Techniques
Cell Line
Cell Proliferation
Cell Separation
Cytotoxicity, Immunologic
Denmark
Female
Flow Cytometry
Humans
Interleukin-2
Interleukin-4
Japan
Male
Microspheres
Middle Aged
Phenotype
Receptors, Antigen, T-Cell, alpha-beta
Recombinant Proteins
Skin
T-Lymphocytes
Time Factors

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title = "In vitro propagation and dynamics of T cells from skin biopsies by methods using interleukins-2 and -4 or anti-CD3/CD28 antibody-coated microbeads",

abstract = "In order to explore the mechanisms of inflammatory skin disorders, we established two methods of expanding skin-derived lymphocytes, one using high levels of interleukin (IL)-2 and IL-4 (method A) and the other using low levels of cytokines and anti-CD3/CD28 microbeads (method B). Both methods provide advantages for functional studies. With either of these two, we could obtain more than 107 cells/ from a 3 mm skin biopsy in 21 days from 23 out of 26 biopsies of various skin diseases. The relevance of these cells was confirmed by shifted T-cell receptor β chain variable region (TCR-Vβ) repertoire and antigen-dependent proliferation in antigen-driven skin disorders. The propagation of skin-resident lymphocytes, seen especially in method A, seems to be mediated by a functional defect of regulatory T cells residing in skin sequentially expanding under the conditions of our methods.",

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author = "Hideo Hashizume and Anker Hansen and Poulsen, {Lars K} and Thomsen, {Allan Randrup} and Masahiro Takigawa and Kristian Thestrup-Pedersen",

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T1 - In vitro propagation and dynamics of T cells from skin biopsies by methods using interleukins-2 and -4 or anti-CD3/CD28 antibody-coated microbeads

AU - Hashizume, Hideo

AU - Hansen, Anker

AU - Poulsen, Lars K

AU - Thomsen, Allan Randrup

AU - Takigawa, Masahiro

AU - Thestrup-Pedersen, Kristian

PY - 2010/9/1

Y1 - 2010/9/1

N2 - In order to explore the mechanisms of inflammatory skin disorders, we established two methods of expanding skin-derived lymphocytes, one using high levels of interleukin (IL)-2 and IL-4 (method A) and the other using low levels of cytokines and anti-CD3/CD28 microbeads (method B). Both methods provide advantages for functional studies. With either of these two, we could obtain more than 107 cells/ from a 3 mm skin biopsy in 21 days from 23 out of 26 biopsies of various skin diseases. The relevance of these cells was confirmed by shifted T-cell receptor β chain variable region (TCR-Vβ) repertoire and antigen-dependent proliferation in antigen-driven skin disorders. The propagation of skin-resident lymphocytes, seen especially in method A, seems to be mediated by a functional defect of regulatory T cells residing in skin sequentially expanding under the conditions of our methods.

AB - In order to explore the mechanisms of inflammatory skin disorders, we established two methods of expanding skin-derived lymphocytes, one using high levels of interleukin (IL)-2 and IL-4 (method A) and the other using low levels of cytokines and anti-CD3/CD28 microbeads (method B). Both methods provide advantages for functional studies. With either of these two, we could obtain more than 107 cells/ from a 3 mm skin biopsy in 21 days from 23 out of 26 biopsies of various skin diseases. The relevance of these cells was confirmed by shifted T-cell receptor β chain variable region (TCR-Vβ) repertoire and antigen-dependent proliferation in antigen-driven skin disorders. The propagation of skin-resident lymphocytes, seen especially in method A, seems to be mediated by a functional defect of regulatory T cells residing in skin sequentially expanding under the conditions of our methods.

KW - Adult

KW - Aged

KW - Antibodies

KW - Antigens, CD28

KW - Antigens, CD3

KW - Biopsy

KW - Cell Culture Techniques

KW - Cell Line

KW - Cell Proliferation

KW - Cell Separation

KW - Cytotoxicity, Immunologic

KW - Denmark

KW - Female

KW - Flow Cytometry

KW - Humans

KW - Interleukin-2

KW - Interleukin-4

KW - Japan

KW - Male

KW - Microspheres

KW - Middle Aged

KW - Phenotype

KW - Receptors, Antigen, T-Cell, alpha-beta

KW - Recombinant Proteins

KW - Skin

KW - T-Lymphocytes

KW - Time Factors

U2 - 10.2340/00015555-0927

DO - 10.2340/00015555-0927

M3 - Journal article

C2 - 20814620

SN - 0001-5555

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SP - 468

EP - 473

JO - Acta Dermato-Venereologica

JF - Acta Dermato-Venereologica

IS - 5

ER -

In vitro propagation and dynamics of T cells from skin biopsies by methods using interleukins-2 and -4 or anti-CD3/CD28 antibody-coated microbeads

Abstract

Keywords

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