Journal of Histochemistry and Cytochemistry Priciples for Free Access to Science
  Search:   
    >> Advanced Search

Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by van der Loos, C. M.
Right arrow Articles by Becker, A. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by van der Loos, C. M.
Right arrow Articles by Becker, A. E.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?
Journal of Histochemistry and Cytochemistry, Vol. 49, 699-710, June 2001, Copyright © 2001, The Histochemical Society, Inc.

ARTICLE

Immunohistochemical Detection of Interferon-{gamma}: Fake or Fact?

Chris M. van der Loosa, Mischa A. Houtkampa, Onno J. de Boera, Peter Teelinga, Allard C. van der Wala, and Anton E. Beckera
a Academic Medical Center, Department of Cardiovascular Pathology, Amsterdam, The Netherlands

Correspondence to: Chris M. van der Loos, Academical Medical Center, Dept. of Cardiovascular Pathology (H0-120), Meibergdreef 9, NL-1105 AZ Amsterdam, The Netherlands. E-mail: c.m.vanderloos@amc.uva.nl

Immunohistochemistry is a widely accepted tool to investigate the presence and immunolocalization of cytokines in tissue sections at the protein level. We have tested the specificity and reproducibility of IFN{gamma} immunohistochemistry on tissue sections with a large panel of anti-IFN{gamma} antibodies. Thirteen different commercially available anti-IFN{gamma} antibodies, including seven advertised and/or regularly applied for immunohistochemistry/-cytochemistry, were tested using a three-step streptavidin–biotin–peroxidase technique and a two-step immunofluorescence (FACS) analysis. Immunoenzyme double staining was used to identify the IFN{gamma}-positive cells. Serial cryostat sections were used of human reactive hyperplastic tonsils, rheumatoid synovium, and inflammatory abdominal aortic aneurysms, known to possess a prominent Th1-type immune response. In vitro phorbol myristate acetate/ionomycin-stimulated T-cells served as positive control; unstimulated cells served as negative control. Cultured T-cells were used adhered to glass slides (immunocytochemistry), in suspension (FACS), or snap-frozen and sectioned (immunohistochemistry). Immunocytochemistry and FACS analysis on stimulated cultured T-cells showed positive staining results with 12 of 13 anti-IFN{gamma} antibodies. However, immunohistochemistry of sectioned stimulated T-cells was negative with all. Unstimulated cells were consistently negative. IFN{gamma} immunohistochemical single- and double staining analysis of the tissue sections showed huge variations in staining patterns, including positivity for smooth muscle cells (n=8), endothelial cells (n=4), extracellular matrix (n=4), and CD138+ plasma cells (n=12). Specific staining of T-cells, as the sole positive staining, was not achieved with any of the 13 antibodies. IFN{gamma}-immunohistochemistry appears unreliable because of lack of specificity to stain T-cells in situ. In fact, depending on the type of anti-IFN{gamma} antibody used, a variety of different cell constituents were nonspecifically stained. Consequently, data based on IFN{gamma}-immunohistochemistry must be interpreted with great caution. (J Histochem Cytochem 49:699–709, 2001)

Key Words: IFN{gamma} protein, immunohistochemistry, immunocytochemistry, T-cells, plasma cells


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 FASEB J.Home page
G. E. Lash, H. A. Otun, B. A. Innes, M. Kirkley, L. De Oliveira, R. F. Searle, S. C. Robson, and J. N. Bulmer
Interferon-{gamma} inhibits extravillous trophoblast cell invasion by a mechanism that involves both changes in apoptosis and protease levels
FASEB J, December 1, 2006; 20(14): 2512 - 2518.
[Abstract] [Full Text] [PDF]

Home page
 CirculationHome page
F. Franceschi, A. R. Sepulveda, A. Gasbarrini, P. Pola, N. G. Silveri, G. Gasbarrini, D. Y. Graham, and R. M. Genta
Cross-Reactivity of Anti-CagA Antibodies With Vascular Wall Antigens: Possible Pathogenic Link Between Helicobacter pylori Infection and Atherosclerosis
Circulation, July 23, 2002; 106(4): 430 - 434.
[Abstract] [Full Text] [PDF]

Home page
 JEMHome page
N. Gerdes, G. K. Sukhova, P. Libby, R. S. Reynolds, J. L. Young, and U. Schonbeck
Expression of Interleukin (IL)-18 and Functional IL-18 Receptor on Human Vascular Endothelial Cells, Smooth Muscle Cells, and Macrophages: Implications for Atherogenesis
J. Exp. Med., January 22, 2002; 195(2): 245 - 257.
[Abstract] [Full Text] [PDF]



Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact
The Journal of Histochemistry & Cytochemistry is owned, published, and licensed by The Histochemical Society © 2001