Separation of human suppressor and helper T cells by concanavalin A-coated sheep erythrocytes.
Normal human peripheral blood mononuclear leukocytes (PBML) were activated by concanavalin A (Con A). Con A-activated and non-activated T cells were separated by E (AET) rosettes (2-aminoethylisothiouronium hydrobromide treated sheep erythrocyte rosettes). Purified T cells were rosetted with Con A-coated sheep red blood cells (Con A-SRBC) at 37 degrees C resulting in Con A-SRBC rosetted and non-rosetted T cells. The Con A-SRBC rosetted T lymphocytes in the T lymphocytes from Con A-activated and non-activated PBML were 44.4 +/- 5.4 percent and 16.0 +/- 7.5 percent (Mean +/- S.D.) while the Con A-SRBC non-rosetted T lymphocytes were 55.6 +/- 5.4 percent and 84.0 +/- 7.5 percent respectively. The Con A-SRBC rosetted and non-rosetted T cells were separated by Ficoll-Hypaque gradient centrifugation. Functional studies of Con A-SRBC rosetted and non-rosetted T cells were performed by in vitro tests using pre-amplified reverse hemolytic plaque assay for measuring numbers of immunoglobulin G (IgG) secreting cells and ELISA quantitation of IgG concentration. Both techniques were used to assess the suppressor and helper functions of the Con A-SRBC rosetted and non-rosetted T cells. The Con A-SRBC rosetted cells obtained from T cells of Con A-activated PBML showed strong suppressor activities to normal PBML in both pre-amplified reverse hemolytic plaque assay and sandwidh ELISA of IgG concentration, while the Con A-SRBC non-rosetted T cells demonstrated strong helper activities to normal PBML in both assay systems.(ABSTRACT TRUNCATED AT 250 WORDS)
Makonkawkeyoon
S
S
Department of Clinical Immunology, Faculty of Associated Medical Sciences, Chiang Mai University, Thailand.
Kasinrerk
W
W
D000818:Animals; D002469:Cell Separation / Q000379:methods; D003208:Concanavalin A; D004797:Enzyme-Linked Immunosorbent Assay; D004912:Erythrocytes / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D012397:Rosette Formation; D012756:Sheep; D006377:T-Lymphocytes, Helper-Inducer / Q000166:cytology / Q000502:physiology*; D050378:T-Lymphocytes, Regulatory / Q000166:cytology / Q000502:physiology*
D007074:Immunoglobulin G; D003208:Concanavalin A