Mouse lymphocytes with and without surface immunoglobulin: preparative scale separation in polystyrene tissue culture dishes coated with specifically purified anti-immunoglobulin

J Immunol Methods. 1977;15(1):47-56. doi: 10.1016/0022-1759(77)90016-3.

Abstract

Mouse spleen cells could be preparatively separated into immunoglobulin positive (Ig+) and immunoglobulin-netative (Ig-)populations by incubating as many as 2 X 10(8) cells per 100 mm diameter petri plate coated with specifically purified goat anti-mouse immunoglobulin. The non-adherent population was 95% or more Ig-, and possessed graft versus host and cytotoxic effector activities, as would be expected for T cells. They could also give a mixed lymphocyte reaction and generate cytotoxic effector activity on culture in vitro. The adherent cells could not be released undamaged from plates coated with undiluted anti-Ig, but they could be released from plates coated with a 1/4 or 1/10 dilution of anti-Ig in an irrelevant antibody. The released cells were over 90% viable by trypan-blue staining, and 94% or more of the viable cells were Ig+.

MeSH terms

  • Animals
  • Antibodies, Anti-Idiotypic / isolation & purification*
  • Antibody Specificity*
  • Binding Sites, Antibody
  • Cell Adhesion
  • Cell Separation
  • Cytotoxicity Tests, Immunologic
  • Goats
  • Graft vs Host Reaction
  • Immune Sera / pharmacology
  • Immunity, Cellular
  • Lymphocyte Culture Test, Mixed
  • Lymphocytes / immunology*
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Polystyrenes*
  • Receptors, Antigen, B-Cell / analysis*
  • Spleen / immunology
  • Thymus Gland / immunology

Substances

  • Antibodies, Anti-Idiotypic
  • Immune Sera
  • Polystyrenes
  • Receptors, Antigen, B-Cell