There is no regulatory role for induced nitric oxide in the regulation of the in vitro proliferative response of bovine mononuclear cells to mitogens, alloantigens or superantigens.
Nitric oxide (NO) is a potent cellular mediator which has been shown to modulate several immune mechanisms. Between species, however, there are considerable differences regarding the signals required for induction of NO as well as the kind of cells capable of producing NO. The object of this study was to determine the kinetics of NO production of bovine blood mononuclear cells (boMNC) stimulated in vitro and to investigate whether it modulates their proliferative response following allogeneic (mixed leukocyte cultures, aMLC), mitogenic (PWM, Con A) or superantigenic (SEA, SEB) stimulation. NO production was indirectly determined with the Griess reagent measuring nitrite (NO2-). Significant but low amounts of NO could be detected as early as day 3 after in vitro stimulation and did noly slightly increase during the 6-8 day culture period. Superantigens (SEA, SEB) and aMLCs (4.3-5.2 microM NO2-) induced a significantly higher nitrite accumulation compared to Con A (2.6 microM NO2-). Generation of nitrite, most likely produced by monocytes/macrophages, could be inhibited by 1 mM N-monomethyl-L-arginine (NMLA). Flow cytometric characterization of various cellular responses revealed no differences between cultures with or without NMLA. This included the determination of blastogenesis, absolute numbers of viable cells, expression density of activation markers (MHC class II, IL-2R alpha) and cellular subpopulations (CD4+, CD8+, sIg+) among blasts. In addition, exogenously provided NO via SNOG in non-toxic concentrations (10(-5)-10(-4) M) did not alter the proliferative reaction of boMNC in vitro. The results suggest that NO is induced after in vitro stimulation of boMNC, however, at a low level, and without having any positive or suppressive effects on the so far tested cellular parameters of activation and proliferation.
Schuberth
H J
HJ
Immunology Unit, School of Veterinary Medicine, Hannover, Germany. jschub@immunologie.tiho-hannover.de
Hendricks
A
A
Leibold
W
W
D000818:Animals; D002417:Cattle; D002455:Cell Division / Q000502:physiology; D004731:Endotoxins / Q000008:administration & dosage; D005260:Female; D007519:Isoantigens; D037102:Lectins / Q000008:administration & dosage; D007963:Leukocytes, Mononuclear / Q000737:chemistry / Q000276:immunology*; D008297:Male; D009569:Nitric Oxide / Q000276:immunology* / Q000378:metabolism; D013211:Staphylococcus aureus / Q000276:immunology; D013601:T-Lymphocytes / Q000737:chemistry / Q000276:immunology*
D004731:Endotoxins; D007519:Isoantigens; D037102:Lectins; D009569:Nitric Oxide