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IgG on Human Blood Lymphocytes Studied by Immunofluorescence
Author(s) -
PETTERSSON D.,
MELLSTEDT H.,
HOLM G.
Publication year - 1978
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.1978.tb00553.x
Subject(s) - immunofluorescence , indirect immunofluorescence , human blood , immunology , antibody , biology , physiology
Surface immunoglobulins (sIg) on human blood lymphocytes were identified by immunofluorescence (IFL) after staining with conjugated F(ab' 2 ) fragments of the anti‐Ig antibodies. A large fraction (≃20%) of freshly isolated lymphocytes was found to carry sIgG. Polyclonal IgG, which was present on Fc‐receptor‐bearing lymphocytes, could be removed by incubation and repeated washings only at 37°C. Lymphocytes treated at 37°C expressed the same percentage of sIg + cells in direct IFL when F(ab') 2 fragments of the antibody was used as when undigested aggregate‐free IgG antibody was used. Indirect IFL using F(ab') 2 fragments in both steps yielded similar sIg + values. However, much higher percentage of cells carried sIg when undigested antibody was included in one of the steps. The results suggest that incubation and washing at 37°C and the use of F(ab) 2 fragments of the anybodies are important to eliminate absorbed sIgG and to avoid absorption of IgG during the staining procedure, thus preventing overestimation of the number of sIg + B lymphocytes identified by IFL.