TY - JOUR
T1 - Cellular immunoadsorbents
T2 - a simplified technique for separation of lymphoid cell populations
AU - Kedar, E.
AU - Ortiz De Landazuri, M.
AU - Bonavida, B.
PY - 1974
Y1 - 1974
N2 - A simplified and efficient technique for separating T lymphoid cells from non T cells of mouse, rat and man was developed. The fractionation procedure is based on the capacity of non T lymphoid cells to bind antigen antibody complexes through an Fc receptor. Lymphoid cells are adsorbed on antibody coated sheep erythrocyte (EA) monolayers prepared in polystyrene Petri dishes previously treated with poly l lysine. Adsorption of spleen cells on these monolayers, assisted by a 5 min centrifugation, resulted in 90 to 98% depletion of the Fc receptor bearing cells (mostly B lymphocytes, but also macrophages and polymorphonuclear leukocytes) in the non adherent cell population. Eighty to 90% of the adherent cells were capable of forming EA rosettes and could be recovered after lysis of the erythrocytes. The non adherent fraction comprised at least 90% T cells. This was determined for mouse and rat spleen cells by lysis with anti thymocyte serum and complement, and by the absence of immunoglobulin bearing cells. The specificity of the adsorption was further substantiated by the following observations: lymphoid cells lacking an Fc receptor were not adsorbed; the adsorption to EA monolayers was inhibited by antigen antibody complexes or by aggregated γ globulin; and only 10 to 15% of the Fc receptor bearing cells were attached to non sensitized erythrocyte (E) monolayers. Fractionation of spleen cells obtained from rats immunized against a syngeneic tumor yielded two separate fractions of cytotoxic lymphoid cells: one non adherent, T cells, mediating direct target cell destruction, and the other (adherent, non T cells) mediating lysis of antibody coated target cells.
AB - A simplified and efficient technique for separating T lymphoid cells from non T cells of mouse, rat and man was developed. The fractionation procedure is based on the capacity of non T lymphoid cells to bind antigen antibody complexes through an Fc receptor. Lymphoid cells are adsorbed on antibody coated sheep erythrocyte (EA) monolayers prepared in polystyrene Petri dishes previously treated with poly l lysine. Adsorption of spleen cells on these monolayers, assisted by a 5 min centrifugation, resulted in 90 to 98% depletion of the Fc receptor bearing cells (mostly B lymphocytes, but also macrophages and polymorphonuclear leukocytes) in the non adherent cell population. Eighty to 90% of the adherent cells were capable of forming EA rosettes and could be recovered after lysis of the erythrocytes. The non adherent fraction comprised at least 90% T cells. This was determined for mouse and rat spleen cells by lysis with anti thymocyte serum and complement, and by the absence of immunoglobulin bearing cells. The specificity of the adsorption was further substantiated by the following observations: lymphoid cells lacking an Fc receptor were not adsorbed; the adsorption to EA monolayers was inhibited by antigen antibody complexes or by aggregated γ globulin; and only 10 to 15% of the Fc receptor bearing cells were attached to non sensitized erythrocyte (E) monolayers. Fractionation of spleen cells obtained from rats immunized against a syngeneic tumor yielded two separate fractions of cytotoxic lymphoid cells: one non adherent, T cells, mediating direct target cell destruction, and the other (adherent, non T cells) mediating lysis of antibody coated target cells.
UR - http://www.scopus.com/inward/record.url?scp=0015976312&partnerID=8YFLogxK
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C2 - 4590702
AN - SCOPUS:0015976312
SN - 0022-1767
VL - 112
SP - 1231
EP - 1243
JO - Journal of Immunology
JF - Journal of Immunology
IS - 3
ER -