Immunocytochemical detection of cytokines and chemokines in Langerhans cells and in vitro derived dendritic cells

Immunocytochemical detection of cytokines and chemokines in Langerhans cells and in vitro derived... We have developed a direct immunocytochemical technique to identify cytokine and chemokine production in epidermal Langerhans cells (LC) and in vitro derived CD14−, CD1a+, CD83+, CD40+ dendritic cells (DC) at the single cell level. Formaldehyde fixation combined with saponin permeabilization preserved cellular morphology and generated a characteristic juxtanuclear staining signal due to the accumulation of cytokine to the Golgi organelle. This approach was used for the assessment of TNF- α , IL-6, IL-8, IL-10, IL-12, GM-CSF, MIP-1 α , MIP-1 β and RANTES producing cells. In contrast, a diffuse cytoplasmic staining was evident for IL-1ra, IL-1 α and IL-1 β production. IL-1ra and IL-1 α were expressed in 10–25% of unstimulated cultured cells, while all the other cytokines were undetectable. IL-1ra, IL-1 α and IL-1 β were also the dominating cytokines, expressed in up to 85% of the DC, after 3 h of LPS stimulation. A significantly lower number of cells (0–5%) synthesized TNF- α , IL-6, IL-10, IL-12 and GM-CSF. The incidence of chemokine producing cells (IL-8, RANTES, MIP-1 α , MIP-1 β ) peaked 10 h after LPS stimulation in up to 60% of the DC. Both immature CD83− and mature CD83+ DC as well as LC had a similar cytokine production pattern. Thus, in comparison to monocytes, LPS stimulation of DC generated a lower incidence of TNF- α , IL-6, IL-10 and IL-12 producing cells while IL-1 was expressed in a comparable number of cells. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Immunological Methods Elsevier

Immunocytochemical detection of cytokines and chemokines in Langerhans cells and in vitro derived dendritic cells

Loading next page...
 
/lp/elsevier/immunocytochemical-detection-of-cytokines-and-chemokines-in-langerhans-nn7H8nRdGT
Publisher
Elsevier
Copyright
Copyright © 1998 Elsevier Ltd
ISSN
0022-1759
DOI
10.1016/S0022-1759(98)00040-4
Publisher site
See Article on Publisher Site

Abstract

We have developed a direct immunocytochemical technique to identify cytokine and chemokine production in epidermal Langerhans cells (LC) and in vitro derived CD14−, CD1a+, CD83+, CD40+ dendritic cells (DC) at the single cell level. Formaldehyde fixation combined with saponin permeabilization preserved cellular morphology and generated a characteristic juxtanuclear staining signal due to the accumulation of cytokine to the Golgi organelle. This approach was used for the assessment of TNF- α , IL-6, IL-8, IL-10, IL-12, GM-CSF, MIP-1 α , MIP-1 β and RANTES producing cells. In contrast, a diffuse cytoplasmic staining was evident for IL-1ra, IL-1 α and IL-1 β production. IL-1ra and IL-1 α were expressed in 10–25% of unstimulated cultured cells, while all the other cytokines were undetectable. IL-1ra, IL-1 α and IL-1 β were also the dominating cytokines, expressed in up to 85% of the DC, after 3 h of LPS stimulation. A significantly lower number of cells (0–5%) synthesized TNF- α , IL-6, IL-10, IL-12 and GM-CSF. The incidence of chemokine producing cells (IL-8, RANTES, MIP-1 α , MIP-1 β ) peaked 10 h after LPS stimulation in up to 60% of the DC. Both immature CD83− and mature CD83+ DC as well as LC had a similar cytokine production pattern. Thus, in comparison to monocytes, LPS stimulation of DC generated a lower incidence of TNF- α , IL-6, IL-10 and IL-12 producing cells while IL-1 was expressed in a comparable number of cells.

Journal

Journal of Immunological MethodsElsevier

Published: May 1, 1998

References

You’re reading a free preview. Subscribe to read the entire article.


DeepDyve is your
personal research library

It’s your single place to instantly
discover and read the research
that matters to you.

Enjoy affordable access to
over 18 million articles from more than
15,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

Search

Query the DeepDyve database, plus search all of PubMed and Google Scholar seamlessly

Organize

Save any article or search result from DeepDyve, PubMed, and Google Scholar... all in one place.

Access

Get unlimited, online access to over 18 million full-text articles from more than 15,000 scientific journals.

Your journals are on DeepDyve

Read from thousands of the leading scholarly journals from SpringerNature, Elsevier, Wiley-Blackwell, Oxford University Press and more.

All the latest content is available, no embargo periods.

See the journals in your area

DeepDyve

Freelancer

DeepDyve

Pro

Price

FREE

$49/month
$360/year

Save searches from
Google Scholar,
PubMed

Create folders to
organize your research

Export folders, citations

Read DeepDyve articles

Abstract access only

Unlimited access to over
18 million full-text articles

Print

20 pages / month

PDF Discount

20% off