Bauhinia purpurea lectin (BPA) binding spectra in hyperplastic human tonsil and in peripheral blood: Immunohistochemical, immunoelectron microscopic, and flow cytometric analyses

A. B. Sarker, T. Akagi, Tadashi Yoshino, K. Fujiwara, I. Murakami

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Abstract

We studied the binding of Bauhinia purpurea lectin (BPA) to human tonsils and peripheral blood mononuclear cells by immunohistochemical, immunoelectron microscopic, and flow cytometric techniques. The effect of several fixatives (acetone, ethanol, buffered and non-buffered formalin, B5, Bodian 11, Bouin's, Carnoy's, Zenker's, and glutaraldehyde) was also examined. BPA was reactive with germinal center lymphocytes, macrophages/histiocytes, follicular dendritic cells, squamous epithelial cells, and a subset of endothelial cells. Mantle zone and paracortical lymphocytes were non-reactive with BPA. The profile of the specific binding characteristic of BPA lectin was found to be influenced by the fixatives. Most significantly, formalin fixation greatly reduced overall binding intensity, particularly making germinal center lymphocytes totally non-reactive. The reaction intensity was most prominent in frozen sections or those fixed in Carnoy's or ethanol solution. The combination of heavy metal salt-containing fixatives with acetic acid usually did not enhance BPA binding. Glutaraldehyde solution used for immunoelectron microscopic study also preserved BPA receptors fairly well, and BPA was confined to the membrane in lymphocytes and to both the membrane and cytoplasm in macrophages/histiocytes and follicular dendritic cells. Neuraminidase treatment of tissues resulted in binding of BPA to lymphocytes that were non-reactive before treatment. Double-staining studies on cell suspensions from tonsils with FITC-BPA and PE-conjugated anti-CD3 or SIg reagents revealed that 28.4% of CD3+ cells and 61.3% of SIg+ cells were BPA reactive. In PBL, 65.6% and 81.4% of CD3+ and SIg+ cells, respectively, were BPA reactive. Neuraminidase treatment also increased the percentage of BPA-reactive lymphocytes. In conclusion, BPA is a marker for macrophages/histiocytes and germinal center lymphocytes, provided that the tissues are unfixed or fixed with suitable fixatives such as ethanol or Carnoy's solution.

Original languageEnglish
Pages (from-to)811-817
Number of pages7
JournalJournal of Histochemistry and Cytochemistry
Volume41
Issue number6
Publication statusPublished - 1993

Fingerprint

Palatine Tonsil
Fixatives
Lymphocytes
Histiocytes
Germinal Center
Follicular Dendritic Cells
Ethanol
Macrophages
Neuraminidase
Glutaral
Bauhinia purpurea lectin
Formaldehyde
Mitogen Receptors
Membranes
Fluorescein-5-isothiocyanate
Frozen Sections
Heavy Metals
Acetone
Lectins
Acetic Acid

Keywords

  • Bauhinia purpurea: Lectin
  • Flow cytometry
  • Human tonsil
  • Immunoelectron microscopy
  • Immunohistochemistry
  • Peripheral blood

ASJC Scopus subject areas

  • Cell Biology
  • Anatomy

Cite this

@article{1a51a89dc1c24fb69e0c906b2e2adf07,
title = "Bauhinia purpurea lectin (BPA) binding spectra in hyperplastic human tonsil and in peripheral blood: Immunohistochemical, immunoelectron microscopic, and flow cytometric analyses",
abstract = "We studied the binding of Bauhinia purpurea lectin (BPA) to human tonsils and peripheral blood mononuclear cells by immunohistochemical, immunoelectron microscopic, and flow cytometric techniques. The effect of several fixatives (acetone, ethanol, buffered and non-buffered formalin, B5, Bodian 11, Bouin's, Carnoy's, Zenker's, and glutaraldehyde) was also examined. BPA was reactive with germinal center lymphocytes, macrophages/histiocytes, follicular dendritic cells, squamous epithelial cells, and a subset of endothelial cells. Mantle zone and paracortical lymphocytes were non-reactive with BPA. The profile of the specific binding characteristic of BPA lectin was found to be influenced by the fixatives. Most significantly, formalin fixation greatly reduced overall binding intensity, particularly making germinal center lymphocytes totally non-reactive. The reaction intensity was most prominent in frozen sections or those fixed in Carnoy's or ethanol solution. The combination of heavy metal salt-containing fixatives with acetic acid usually did not enhance BPA binding. Glutaraldehyde solution used for immunoelectron microscopic study also preserved BPA receptors fairly well, and BPA was confined to the membrane in lymphocytes and to both the membrane and cytoplasm in macrophages/histiocytes and follicular dendritic cells. Neuraminidase treatment of tissues resulted in binding of BPA to lymphocytes that were non-reactive before treatment. Double-staining studies on cell suspensions from tonsils with FITC-BPA and PE-conjugated anti-CD3 or SIg reagents revealed that 28.4{\%} of CD3+ cells and 61.3{\%} of SIg+ cells were BPA reactive. In PBL, 65.6{\%} and 81.4{\%} of CD3+ and SIg+ cells, respectively, were BPA reactive. Neuraminidase treatment also increased the percentage of BPA-reactive lymphocytes. In conclusion, BPA is a marker for macrophages/histiocytes and germinal center lymphocytes, provided that the tissues are unfixed or fixed with suitable fixatives such as ethanol or Carnoy's solution.",
keywords = "Bauhinia purpurea: Lectin, Flow cytometry, Human tonsil, Immunoelectron microscopy, Immunohistochemistry, Peripheral blood",
author = "Sarker, {A. B.} and T. Akagi and Tadashi Yoshino and K. Fujiwara and I. Murakami",
year = "1993",
language = "English",
volume = "41",
pages = "811--817",
journal = "Journal of Histochemistry and Cytochemistry",
issn = "0022-1554",
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TY - JOUR

T1 - Bauhinia purpurea lectin (BPA) binding spectra in hyperplastic human tonsil and in peripheral blood

T2 - Immunohistochemical, immunoelectron microscopic, and flow cytometric analyses

AU - Sarker, A. B.

AU - Akagi, T.

AU - Yoshino, Tadashi

AU - Fujiwara, K.

AU - Murakami, I.

PY - 1993

Y1 - 1993

N2 - We studied the binding of Bauhinia purpurea lectin (BPA) to human tonsils and peripheral blood mononuclear cells by immunohistochemical, immunoelectron microscopic, and flow cytometric techniques. The effect of several fixatives (acetone, ethanol, buffered and non-buffered formalin, B5, Bodian 11, Bouin's, Carnoy's, Zenker's, and glutaraldehyde) was also examined. BPA was reactive with germinal center lymphocytes, macrophages/histiocytes, follicular dendritic cells, squamous epithelial cells, and a subset of endothelial cells. Mantle zone and paracortical lymphocytes were non-reactive with BPA. The profile of the specific binding characteristic of BPA lectin was found to be influenced by the fixatives. Most significantly, formalin fixation greatly reduced overall binding intensity, particularly making germinal center lymphocytes totally non-reactive. The reaction intensity was most prominent in frozen sections or those fixed in Carnoy's or ethanol solution. The combination of heavy metal salt-containing fixatives with acetic acid usually did not enhance BPA binding. Glutaraldehyde solution used for immunoelectron microscopic study also preserved BPA receptors fairly well, and BPA was confined to the membrane in lymphocytes and to both the membrane and cytoplasm in macrophages/histiocytes and follicular dendritic cells. Neuraminidase treatment of tissues resulted in binding of BPA to lymphocytes that were non-reactive before treatment. Double-staining studies on cell suspensions from tonsils with FITC-BPA and PE-conjugated anti-CD3 or SIg reagents revealed that 28.4% of CD3+ cells and 61.3% of SIg+ cells were BPA reactive. In PBL, 65.6% and 81.4% of CD3+ and SIg+ cells, respectively, were BPA reactive. Neuraminidase treatment also increased the percentage of BPA-reactive lymphocytes. In conclusion, BPA is a marker for macrophages/histiocytes and germinal center lymphocytes, provided that the tissues are unfixed or fixed with suitable fixatives such as ethanol or Carnoy's solution.

AB - We studied the binding of Bauhinia purpurea lectin (BPA) to human tonsils and peripheral blood mononuclear cells by immunohistochemical, immunoelectron microscopic, and flow cytometric techniques. The effect of several fixatives (acetone, ethanol, buffered and non-buffered formalin, B5, Bodian 11, Bouin's, Carnoy's, Zenker's, and glutaraldehyde) was also examined. BPA was reactive with germinal center lymphocytes, macrophages/histiocytes, follicular dendritic cells, squamous epithelial cells, and a subset of endothelial cells. Mantle zone and paracortical lymphocytes were non-reactive with BPA. The profile of the specific binding characteristic of BPA lectin was found to be influenced by the fixatives. Most significantly, formalin fixation greatly reduced overall binding intensity, particularly making germinal center lymphocytes totally non-reactive. The reaction intensity was most prominent in frozen sections or those fixed in Carnoy's or ethanol solution. The combination of heavy metal salt-containing fixatives with acetic acid usually did not enhance BPA binding. Glutaraldehyde solution used for immunoelectron microscopic study also preserved BPA receptors fairly well, and BPA was confined to the membrane in lymphocytes and to both the membrane and cytoplasm in macrophages/histiocytes and follicular dendritic cells. Neuraminidase treatment of tissues resulted in binding of BPA to lymphocytes that were non-reactive before treatment. Double-staining studies on cell suspensions from tonsils with FITC-BPA and PE-conjugated anti-CD3 or SIg reagents revealed that 28.4% of CD3+ cells and 61.3% of SIg+ cells were BPA reactive. In PBL, 65.6% and 81.4% of CD3+ and SIg+ cells, respectively, were BPA reactive. Neuraminidase treatment also increased the percentage of BPA-reactive lymphocytes. In conclusion, BPA is a marker for macrophages/histiocytes and germinal center lymphocytes, provided that the tissues are unfixed or fixed with suitable fixatives such as ethanol or Carnoy's solution.

KW - Bauhinia purpurea: Lectin

KW - Flow cytometry

KW - Human tonsil

KW - Immunoelectron microscopy

KW - Immunohistochemistry

KW - Peripheral blood

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