Decrease of intracellular fluorescein fluorescence polarization (IFFP) in human peripheral blood lymphocytes undergoing stimulation with phytohaemagglutinin (PHA), concanavalin A (ConA), pokeweed mitogen (PWM) and anti-CD3 antibody.
In the present study we describe the induction of changes in intracellular fluorescein fluorescence polarization (IFFP) in lymphocytes undergoing activation with a variety of stimulants. These stimulants included the lectins phytohaemagglutinin (PHA), concanavalin (ConA), pokeweed mitogen (PWM) and anti-CD3 antibody. Changes in IFFP were detected in individual cells using the Cellscan apparatus. Our results show that by employing mitogenic concentrations of PHA, as revealed in a [3H]-thymidine incorporation assay, a decrease in the IFFP in human peripheral blood lymphocytes (PBL) occurred within 40 min. ConA and anti-CD3 affected similarly IFFP, whereas PWM, a B lymphocyte lectin, had no effect on IFFP at the concentrations employed. Kinetic analysis revealed that changes in IFFP occurred within 20-40 min after exposure to the stimulants and lasted for 24 h. Our results show that stimulants which activate CD3+ lymphocytes caused immediate changes in IFFP, in an enriched population of human PBL. The possible mechanisms involved in IFFP modulation following exposure to selected stimulants are discussed.
Sourasky Medical Center, Tel-Aviv, Israel.
A
A
Eisenthal
O
O
Marder
D
D
Dotan
S
S
Baron
B
B
Lifschitz-Mercer
S
S
Chaitchik
R
R
Tirosh
A
A
Weinreb
M
M
Deutsch
D000918:Antibody Specificity; D017252:CD3 Complex / Q000276:immunology*; D003208:Concanavalin A / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D019793:Fluorescein; D005452:Fluoresceins; D005454:Fluorescence Polarization; D006801:Humans; D007700:Kinetics; D037102:Lectins / Q000494:pharmacology*; D008213:Lymphocyte Activation / Q000187:drug effects / Q000502:physiology*; D008214:Lymphocytes / Q000502:physiology / Q000648:ultrastructure; D008855:Microscopy, Electron, Scanning; D010835:Phytohemagglutinins / Q000494:pharmacology; D011043:Pokeweed Mitogens / Q000494:pharmacology
D017252:CD3 Complex; D005452:Fluoresceins; D037102:Lectins; D010835:Phytohemagglutinins; D011043:Pokeweed Mitogens; D003208:Concanavalin A; D019793:Fluorescein