title: Simultaneous detection of T lymphocyte-related antigens (CD4/CD8, CD57, TCRβ) with nuclei by fluorescence-based immunohistochemistry in paraffin-embedded human lymph node, liver cancer and stomach cancer.
abstract: OBJECTIVE
  To develop a method for immunohistochemical staining of three different T lymphocyte antigens (CD4 or CD8, CD57 and TCRβ) on the same tissue section and to determine whether tissues have been infiltrated with T lymphocytes expressing these markers.
  
  
  METHODS
  Commercially available antibodies were tested for immunohistochemical usefulness in a dye-based conventional single-immunostaining method after antigen retrieval on paraffin-embedded human lymph nodes. We searched for the combination of antibodies that could detect T lymphocyte antigens on the same section without any cross-reactivity and that have fluorescent signals robust enough to overcome paraffin autofluorescence.
  
  
  RESULTS
  Application of the antigen retrieval technique and the Sudan black B quenching technique enabled staining of paraffin-embedded tissue sections with fluorescent-labeled secondary antibodies. The combination of primary and secondary antibodies that could simultaneously detect the T lymphocyte antigens CD4 or CD8, CD57 and TCRβ in histochemical analysis of a paraffin-embedded human lymph node section was established, and was successfully applied to a human tissue section infiltrated with T lymphocytes that express these markers.
  
  
  CONCLUSIONS
  The antibodies listed here would be helpful for histopathologists who wish to investigate T lymphocytes in the paraffin-embedded sections that have accumulated in pathology labs throughout the world.
authors.affiliation: Department of Health Science, Ishikawa Prefectural Nursing University, Kahoku, Ishikawa, Japan. tnakata1@mail.goo.ne.jp
authors.forename: Takahiro
authors.initials: T
authors.lastname: Nakata
authors.forename: Eiji
authors.initials: E
authors.lastname: Takayama
authors.forename: Hirohito
authors.initials: H
authors.lastname: Magari
authors.forename: Jun
authors.initials: J
authors.lastname: Kato
authors.forename: Nobuo
authors.initials: N
authors.lastname: Kondo
authors.forename: Ichinose
authors.initials: I
authors.lastname: Masao
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D015704:CD4 Antigens / Q000032:analysis*; D018959:CD57 Antigens / Q000032:analysis*; D016827:CD8 Antigens / Q000032:analysis*; D005453:Fluorescence; D006801:Humans; D007124:Immunoenzyme Techniques; D008113:Liver Neoplasms / Q000276:immunology* / Q000473:pathology; D008198:Lymph Nodes / Q000276:immunology; D016612:Paraffin Embedding; D011379:Prognosis; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000032:analysis*; D013274:Stomach Neoplasms / Q000276:immunology* / Q000473:pathology; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D018959:CD57 Antigens; D016827:CD8 Antigens; D016693:Receptors, Antigen, T-Cell, alpha-beta