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DENSITY SEPARATION AND CHARACTERISATION OF HUMAN BLOOD LEUCOCYTES, INCLUDING T AND B LYMPHOCYTES
Author(s) -
Williams N,
Moore MAS,
Shortman K,
Condon Linda,
Pike Beverley,
Nossal GJV
Publication year - 1974
Publication title -
australian journal of experimental biology and medical science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.999
H-Index - 104
eISSN - 1440-1711
pISSN - 0004-945X
DOI - 10.1038/icb.1974.48
Subject(s) - phytohaemagglutinin , peripheral blood mononuclear cell , biology , surface immunoglobulin , antibody , immunology , microbiology and biotechnology , lymphocyte , b cell , in vitro , biochemistry
Summary Human blood leucocytes were separated on the basis of buoyant density on high resolution, continuous albumin gradients. Polymorphonuclear leucocytes were dense, while mononuclear cells were light. There was substantial overlap between monocytes and lymphocytes. Both populations were heterogeneous within a restricted density range, and both were variable from one individual to another. This heterogeneity and variability of mononuclear elements contrasted with other human blood cells, such as erythrocytes and the progenitors of monocytes and granulocytes. The possible basis for variability is discussed. There was some separation between T and B lymphocytes, with the B cells (cells with a high surface density of immunoglobulin) enriched in the lighter density fractions. However, the overlap between T and B cells was too extensive for use as a general separation procedure. The response to phytohaemagglutinin mitogen (PHA) appeared to follow the overall T cell (immunoglobulin‐negative) distribution, with no marked separation of PHA responsive from PHA non‐responsive lymphocytes.