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Mitogenic Activation of Human Lymphocytes: a Protein A Plaque Assay Evaluation of Polyclonal B‐Cell Activators
Author(s) -
HAMMARSTRÖM L.,
BIRD A. G.,
SMITH C. I. E.
Publication year - 1980
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.1365-3083.1980.tb00202.x
Subject(s) - polyclonal antibodies , virus quantification , antibody , microbiology and biotechnology , lipopolysaccharide , biology , antigen , b cell , immunology , immunoglobulin m , immunoglobulin g , spleen , virus
Using the protein A plaque assay, the capacity of various polyclonal B cell activators to induce differentiation in human B lymphocytes was investigated. Dextran sulphate and native Dextran were both virtually devoid of mitogenic properties, Lipopolysaccharide, however, was round to be a potent mitogen in human cells that, although giving rise to low DNA synthetic response, induced high numbers of immunoglobulin‐synthesizing cells. Mean plaque‐forming cell (PFC) numbers in healthy blood donors assayed on the optimal day (days 5–7) were 23, 493 IgM/10 4 cells, 11, 288 IgG/10 6 cells, and 2643 IgA/10 6 cells. Values obtained in spleen cells, peaking at days 4–6, were slightly higher. Purified protein derivative (PPD) was equally or oven more‐effective than lipopolysaccharide (LPS) in generating PFC of different subclasses in peripheral blood with mean of 29, 241 IgM/10 6 , 21, 269 IgG/10 6 , and 3681 IgA/10 6 . PPD furthermore induced a marked DNA synthetic response in human lymphocytes. These data suggest that LPS and PPD may both be used as functional markers in human cells when analysing patients with a suspected immunodeficiency state. It is suggested that cultures should be assayed using the protein A plaque assay, thereby being able not only to investigate the individual immunoglobulin classes but also to avoid the possible hazards involved in measuring antigen‐specific responses in patients whose prior immunization to the antigen tested can never be totally excluded