Evaluation of CD5 and other differentiation antigens on human immunoglobulin-secreting cells using a combination of immunobead rosetting and reverse haemolytic plaque formation.
This paper describes a new method, with high specificity and sensitivity, for evaluating cell surface markes such as differentiation antigens and cytokine receptors on immunoglobulin-secreting cells. Mononuclear cells, freshly derived from peripheral blood or following stimulation in vitro with pokeweed mitogen or Staphylococcus aureus Cowan I, are partially depleted of T cells and monocytes using immunomagnetic beads (Dynabeads) coated with anti-CD2. The cells are incubated with Dynabeads coated with monoclonal antibody against the cell marker under investigation and then used in a protein A haemolytic plaque assay. Plaque-forming cells (PFC) with (marker-positive) and without (marker-negative) attached beads can be readily enumerated. Values are given for percentages of IgG-, IgA- and IgM-PFC bearing CD19, CD38, CD25 and CD5.
Jones
B M
BM
Department of Pathology, University of Hong Kong.
D036541:ADP-ribosyl Cyclase; D051997:ADP-ribosyl Cyclase 1; D000328:Adult; D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000032:analysis*; D018941:Antigens, CD19; D000943:Antigens, Differentiation / Q000032:analysis*; D000944:Antigens, Differentiation, B-Lymphocyte / Q000032:analysis; D001402:B-Lymphocytes / Q000032:analysis*; D018956:CD5 Antigens; D006462:Hemolytic Plaque Technique; D006801:Humans; D008562:Membrane Glycoproteins; D008875:Middle Aged; D015375:Receptors, Interleukin-2 / Q000032:analysis; D012397:Rosette Formation
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D018941:Antigens, CD19; D000943:Antigens, Differentiation; D000944:Antigens, Differentiation, B-Lymphocyte; D018956:CD5 Antigens; D008562:Membrane Glycoproteins; D015375:Receptors, Interleukin-2; D036541:ADP-ribosyl Cyclase; C498154:CD38 protein, human; D051997:ADP-ribosyl Cyclase 1