Electrophoretic fractionation of murine lymphoid cells. I. Enrichment of peripheral T lymphocyte subsets with distinct surface phenotypes.
The purpose of this work was to evaluate the efficiency of free-flow electrophoresis as a method for separating mouse lymphocyte subsets. The surface phenotype of the cells contained in the various fractions collected after electrophoresis of CBA/J lymph node cells was investigated by means of single- and 2-color flow cytofluorometry (FCF) analysis. In agreement with previous works, B cells (sIg+, Thy-1-) were found to segregate in the low mobility (LM) fractions and T cells (sIg-, Thy-1+) in the high-mobility (HM) fractions. While the mean fluorescence intensity of sIg staining did not significantly vary as a function of electrophoretic mobility (EPM) that of Thy-1 staining tended to decrease with increasing EPM. The distribution of Lyt-1+ cells was roughly parallel to that of Thy-1+ cells. However, 2-color FCF analysis suggested the existence, in addition to a major Thy-1+ Lyt-1+ subpopulation, of a minor subset of Thy-1- Lyt-1+ cells. Lyt-2+ cells made up a peak in the cathodic HM region where they were enriched by up to 3-fold, and a trail in the more anodic HM fractions. Two-color FCF analysis showed that all Lyt-2+ cells recovered in these various electrophoretic fractions expressed the Lyt-1 antigen. Taken together, these data demonstrate that free-flow electrophoresis provides a powerful tool for the delineation and substantial enrichment of phenotypically distinct mouse peripheral T cell subsets.
D000818:Animals; D000950:Antigens, Ly / Q000235:genetics; D000954:Antigens, Surface / Q000302:isolation & purification; D005434:Flow Cytometry; D008221:Lymphoid Tissue / Q000166:cytology*; D051379:Mice; D008808:Mice, Inbred CBA; D010641:Phenotype; D013601:T-Lymphocytes / Q000145:classification*; D018800:Thy-1 Antigens
D000950:Antigens, Ly; D000954:Antigens, Surface; D018800:Thy-1 Antigens