Sustained increases in cytosolic calcium during T lymphocyte allosensitization, proliferation, and acquisition of locomotor function.
Activation of resting T cells is accompanied by an increase in cytosolic calcium ([Ca2+]i). However, the role of [Ca2+]i in the effector function of allosensitized cells, and how this may affect the evolution of the allograft response is unknown. To evaluate this more directly, we determined [Ca2+]i in both unsensitized T cells (C57BL/6 murine thymocytes) and in allosensitized T cells derived from different days of a C57BL/6 anti-DBA/2J mixed leukocyte culture. To correlate potential changes in [Ca2+]i with concomitant development of T cell effector function, the [Ca2+]i, proliferation (3HTdR uptake), and random locomotion (in vitro modified Boyden chamber assay) of the same cells was assayed simultaneously. Allosensitized T cells exhibited higher (P less than 0.05) [Ca2+]i than unsensitized thymocytes on all days of culture tested. Further, there was a progressive rise in [Ca2+]i during the course of allosensitization. Con A stimulated an increase in [Ca2+]i over basal levels (P less than .05) for all cell types. A rise in [Ca2+]i preceded the onset of maximal allosensitized T cell proliferation (which peaked at day 7) and this continued to increase even after completion of DNA synthesis. In contrast, optimal T cell locomotion coincided with maximal [Ca2+]i, well after cell division had occurred. Prostaglandin E2, a known inhibitor of lymphocyte function, did not alter either basal or Con A-stimulated [Ca2+]i in thymocytes or MLC cells. These results indicate that [Ca2+]i signaling persists long after initial T lymphocyte alloactivation, and is maintained during DNA synthesis and acquisition of locomotor capacity. Furthermore, the inhibitory effects of PGE2 on allosensitized T lymphocyte function may be mediated by a calcium-independent mechanism.
Jordan
M L
ML
Department of Surgery, University of Pittsburgh, Pennsylvania 15213.
Vidgen
D F
DF
Wright
J
J
Odell
M
M
Mills
G B
GB
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002455:Cell Division; D002465:Cell Movement; D002478:Cells, Cultured; D003208:Concanavalin A / Q000494:pharmacology; D003600:Cytosol / Q000378:metabolism; D015232:Dinoprostone / Q000494:pharmacology; D066298:In Vitro Techniques; D007519:Isoantigens / Q000276:immunology; D008213:Lymphocyte Activation; D007959:Lymphocyte Culture Test, Mixed; D051379:Mice; D008815:Mice, Inbred Strains; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D013950:Thymus Gland / Q000166:cytology
D007519:Isoantigens; D003208:Concanavalin A; D015232:Dinoprostone; D002118:Calcium