This Article Full Text Full Text (PDF) All Versions of this Article: jhc.2008.952259v1 57/2/101    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Mizutani, Y. Articles by Tsutsumi, Y. Search for Related Content PubMed PubMed Citation Articles by Mizutani, Y. Articles by Tsutsumi, Y. Social Bookmarking                         What's this?

Enzyme-labeled Antigen Method: Histochemical Detection of Antigen-specific Antibody-producing Cells in Tissue Sections of Rats Immunized With Horseradish Peroxidase, Ovalbumin, or Keyhole Limpet Hemocyanin

Yasuyoshi Mizutani, Shinya Tsuge, Kazuya Shiogama, Ryoichi Shimomura, Shingo Kamoshida, Ken-ichi Inada and Yutaka Tsutsumi

Department of Pathology, Fujita Health University School of Medicine, Toyoake, Japan

Correspondence to: Yutaka Tsutsumi, MD, Department of Pathology, Fujita Health University School of Medicine, Toyoake, Aichi 470-1192, Japan. E-mail: tsutsumi{at}fujita-hu.ac.jp

The enzyme-labeled antigen method is a histochemical technique that visualizes antigen-specific antibody-producing cells in tissue sections, originally documented in 1968. In this study, we attempted to reemerge this hidden but potentially useful method in rat models immunized with horseradish peroxidase (HRP), ovalbumin (OA), or keyhole limpet hemocyanin (KLH). After repeated immunization in footpads, popliteal, groin, and axillary lymph nodes and spleen were sampled. Paraformaldehyde-prefixed frozen sections were incubated with HRP, biotinylated OA, or biotinylated KLH. Proteinase K pretreatment and the secondary use of HPR-labeled streptavidin were applied in the latter two situations. Plasma cells producing antigen-specific antibodies were visualized. Proportions of antigen-specific antibody-producing cells in total plasma cells shown with the immunoperoxidase method for rat immunoglobulins were evaluated. The percentage of antigen-specific plasma cells reached 50% of total plasma cells in the regional lymph nodes. The specificity was confirmed by (a) negativity in non-immune rat tissue, (b) negativity with indifferent antigen probes, and (c) abolishment of the reactivity with the corresponding rat serum. In buffered formalin-fixed, paraffin-embedded tissues, fewer plasma cells were labeled for HRP and KLH antibody reactivity after strong proteolysis and prolonged incubation. Expectedly, this method allows us to observe antigen-specific antibody-producing cells under varied pathological conditions. (J Histochem Cytochem 57:101–111, 2009)

Key Words: enzyme-labeled antigen method • horseradish peroxidase • ovalbumin • keyhole limpet hemocyanin • antigen-specific antibody-producing cells

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

Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact

The Journal of Histochemistry & Cytochemistry is owned, published, and licensed by The Histochemical Society © 2009