Premium
The separation of different cell classes from lymphoid organs. X. Preparative electrophoretic separation of lymphocyte sub‐populations from mouse spleen and thoracic duct lymph
Author(s) -
v. Boehmer H.,
Shortman K.,
Nossal G. J. V.
Publication year - 1974
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1040830209
Subject(s) - spleen , surface immunoglobulin , lymph , biology , microbiology and biotechnology , lymphocyte , b cell , thoracic duct , lymphatic system , antigen , electrophoresis , immunology , antibody , pathology , medicine , biochemistry
Conditions have been established for the separation of viable mouse lymphoid cells by continuous free‐buffer film preparative electrophoresis. The detailed electrophoretic distribution profiles of T and B lymphocytes from mouse spleen and thoracic duct have been determined. Cell surface θ‐antigen was used as a marker for T cells, and high surface‐density of immunoglobulin as a marker for B cells. Spleen cells from athymic “nude” mice were also studied. In the unselected normal spleen cell populations B lymphocytes are heterogeneous, about 60% being of low mobility with the remainder distributing broadly, and extending into the highest mobility fractions. T lymphocytes are predominantly of high mobility. Lymphoid cells lacking markers of either the B or T lineage are of intermediate mobility. There is only partial separation of T and B cells because of the extensive overlap between the populations. The high mobility B cells, which separate along with T cells, include a substantial proportion of large cells, and include cells with high surface density of immunoglobulin. The majority of these large B cells can be selectively eliminated by their adherence on passage through a glass‐bead column. By pre‐selecting the 50% non‐adherent lymphocytes from spleen as the starting material, a very sharp and more extensive separation of B and T cells can be achieved, with 100% pure B cells and 90% pure T cells in many fractions. However these samples are not representative of the total T and B cell populations of spleen. In thoracic duct lymph high mobility B‐cells are absent, there is little overlap between T and B cell mobility. 100% pure T and B cells can be isolated.