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flow cytometry Boyden chamber technique
9
Patients
Cell isolation and culture
Chemotaxis assay
healthy donors and patients with AD or psoriasis
Peripheral blood mononuclear cells (PBMCs) were isolated by Lymphoprep densitygradient centrifugation from healthy donors and from patients with AD and psoriasis
Results
Discussion
.
2
Introduction
keywords
background
present study
purpose
.
3
Keywords
• atopic dermatitis; (特应性皮炎) • chemokines; （趋化因子） • human macrophages; （人巨噬细胞） • IP-10/CXCL10; • MDC/CCL22; • psoriasis; （牛皮癣；银屑病） • Staphylococcus aureus; （金黄色葡萄球菌） • α-toxin.（α-毒素）
•Distinct expression of chemokine (C-C motif) ligand (配体 CCL) and chemokine (C-X-C motif) ligand (配体CXCL) chemokines has been documented in both diseases.
IFN-c-induced protein of 10-kDa (IP-10)/CXCL10
macrophage-derived chemokine (MDC)/CCL22
Cell surface markers’ expression and chemotaxis
.
qRT–PCR or ELISA
we studied the effects of α-toxin on Th1- and Th2related chemokines in macrophages from patients with AD and psoriasis where the intrinsic abnormal and different chemokines production profile is well defined.
Macrophages from patients with atopic dermatitis show a reduced CXCL10 expression in response to staphylococcal a-toxin.
.
1
Introduction
Materials and Methods
.
6
Purpose
The effects of α-toxin on human macrophages still remain unclear.
We show here for the first time that α-toxin induces the Th1-related chemokine CXCL10 in human macrophages.
•the effects of a-toxin on human macrophages in patients
with AD and psoriasis?
.
5
Present study
Recent studies demonstrated that infiltration of inflammatory cells into tissues is regulated by chemokines. Two main subfamilies, recently renamed chemokine (C-C motif) ligand (CCL) and chemokine (C-X-C motif) ligand (CXCL) chemo-kines, have been distinguished .
The chemotactic activity of supernatants from
a-toxin stimulated macrophages on
lymphocytes (CD4+ T cells) was determined
using a B. oyden chamber technique
.
7
Materials and Methods
Patients
Materials and Methods
Cytokine assessment by ELISA
Western blot
Cell isolation and culture
Statistical analysis .
8
Methods
.
4
Background:
•Patients with atopic dermatitis (AD) and psoriasis are frequently colonized with Staphylococcus aureus (S. aureusຫໍສະໝຸດ Baidu. one-third of them producing α-toxin, which is cor-related with the severity of eczema in AD.
Several studies have suggested a crucial involvement of CCL chemokines in inflammation.
Chemokines and their receptors are implicated in the development of symptoms of AD and psoriasis .CXCL10 plays a role in pathogenesis of psoriasis .
9
Patients
Cell isolation and culture
Chemotaxis assay
healthy donors and patients with AD or psoriasis
Peripheral blood mononuclear cells (PBMCs) were isolated by Lymphoprep densitygradient centrifugation from healthy donors and from patients with AD and psoriasis
Results
Discussion
.
2
Introduction
keywords
background
present study
purpose
.
3
Keywords
• atopic dermatitis; (特应性皮炎) • chemokines; （趋化因子） • human macrophages; （人巨噬细胞） • IP-10/CXCL10; • MDC/CCL22; • psoriasis; （牛皮癣；银屑病） • Staphylococcus aureus; （金黄色葡萄球菌） • α-toxin.（α-毒素）
•Distinct expression of chemokine (C-C motif) ligand (配体 CCL) and chemokine (C-X-C motif) ligand (配体CXCL) chemokines has been documented in both diseases.
IFN-c-induced protein of 10-kDa (IP-10)/CXCL10
macrophage-derived chemokine (MDC)/CCL22
Cell surface markers’ expression and chemotaxis
.
qRT–PCR or ELISA
we studied the effects of α-toxin on Th1- and Th2related chemokines in macrophages from patients with AD and psoriasis where the intrinsic abnormal and different chemokines production profile is well defined.
Macrophages from patients with atopic dermatitis show a reduced CXCL10 expression in response to staphylococcal a-toxin.
.
1
Introduction
Materials and Methods
.
6
Purpose
The effects of α-toxin on human macrophages still remain unclear.
We show here for the first time that α-toxin induces the Th1-related chemokine CXCL10 in human macrophages.
•the effects of a-toxin on human macrophages in patients
with AD and psoriasis?
.
5
Present study
Recent studies demonstrated that infiltration of inflammatory cells into tissues is regulated by chemokines. Two main subfamilies, recently renamed chemokine (C-C motif) ligand (CCL) and chemokine (C-X-C motif) ligand (CXCL) chemo-kines, have been distinguished .
The chemotactic activity of supernatants from
a-toxin stimulated macrophages on
lymphocytes (CD4+ T cells) was determined
using a B. oyden chamber technique
.
7
Materials and Methods
Patients
Materials and Methods
Cytokine assessment by ELISA
Western blot
Cell isolation and culture
Statistical analysis .
8
Methods
.
4
Background:
•Patients with atopic dermatitis (AD) and psoriasis are frequently colonized with Staphylococcus aureus (S. aureusຫໍສະໝຸດ Baidu. one-third of them producing α-toxin, which is cor-related with the severity of eczema in AD.
Several studies have suggested a crucial involvement of CCL chemokines in inflammation.
Chemokines and their receptors are implicated in the development of symptoms of AD and psoriasis .CXCL10 plays a role in pathogenesis of psoriasis .