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Identification of mononuclear cells in human blood. II. Evaluation of morphological and immunological aspects of native and formaldehyde-fixed cell populations

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Author: Schuit, H.R.E. · Hijmans, W.
Type:article
Date:1980
Institution: Instituut voor experimentele gerontologie TNO
Source:Clinical and Experimental Immunology, 3, 41, 567-574
Identifier: 228879
Keywords: Biology · Fc receptor · Lymphocyte antibody · Blood and hemopoietic system · Cytology · Human cell · Immunofluorescence · Lymphocyte subpopulation · Methodology · Antigen-Antibody Complex · Blood Preservation · Fluorescent Antibody Technique · Formaldehyde · Immunoglobulins, Surface · Lymphocytes · Microscopy, Phase-Contrast · Monocytes · Receptors, Fc

Abstract

The presence of surface-associated immunoglobulins and Fc receptors on mononuclear cells from normal human blood waas investigated by the direct immunofluorescence technique combined with phase-contrast microscopy. Formaldehyde-fixed cells were compared to unfixed cells and to cells preincubated at 37??C. In the unfixed samples a separate population which showed Fc receptors in an immunofluorescence technique using a labelled antigen-antibody complex was detected. This cell population showed an atypical, i.e. not clearly membrane-associated, pattern of fluorescence with anti-Fab conjugates. This interaction most probably is due to autologous IgG molecules taken up by these cells from the donor serum. Using phase-contrast microscopy, these cells were morphologically distinct from lymphocytes and mature monocytes. They will be referred to as 'undefined mononuclear cells' (UMC). After formaldehyde fixation or preincubation at 37??C the interaction of the UMC with anti-Fab conjugates could no longer be demonstrated. Mature monocytes show the same atypical fluorescence pattern with anti-Fab conjugates, but in contrast to the UMC the interaction persits after formaldehyde fixation or preincubation at 37??C. No evidence was found for passive uptake of labelled IgG from conjugates by any mononuclear cell. F(ab')2 fragments of IgG from antisera gave results similar to those obtained with intact IgG fractions. The morphology of the different cell subpopulations is described and their relative numbers in normal blood are given. Formaldehyde fixation proved to be a simple and useful procedure, especially for the determination of the number of B lymphocytes, because the Fc receptor of the undefined mononuclear cells does not give rise to confusion. Chemicals/CAS: Antigen-Antibody Complex; Formaldehyde, 50-00-0; Immunoglobulins, Surface; Receptors, Fc