Methods, compositions, and kits for the treatment of medical conditions
a technology for medical conditions and compositions, applied in the field of immunoinflammatory disorders and ophthalmic disorders, can solve the problems of rubbing of bones against each other, and affecting so as to achieve less toxic effects, improve the effect of treatment effect, and reduce the effect of toxic side effects
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[0339] The effects of test compound combinations on TNFα secretion were assayed in white blood cells from human buffy coat stimulated with lipopolysaccharide or phorbol 12-myristate 13-acetate (PMA) and ionomycin as follows. The results from these experiments are set forth in FIGS. 1A-1MM.
[0340] Lipopolysaccharide (LPS)
[0341] A 100 μl suspension of diluted human white blood cells contained within each well of a polystyrene 384-well plate (NalgeNunc) was stimulated to secrete TNFα by treatment with a final concentration of 2 μg / mL lipopolysaccharide (Sigma L-4130). Various concentrations of each test compound were added at the time of stimulation. After 16-18 hours of incubation at 37° C. in a humidified incubator, the plate was centrifuged and the supernatant transferred to a white opaque polystyrene 384-well plate (NalgeNunc, Maxisorb) coated with an anti-TNFα antibody (PharMingen, #551220). After a two-hour incubation, the plate washed (Tecan PowerWasher 384) with PBS containing...
example 2
[0344] Both corticosteroids and NsIDIs suppress cytokine production in cell culture models of immune function. We tested the effect of the combination of various concentrations of NsIDIs and corticosteroids on cytokine production in a cell culture model of immune function. We propose that combinations that demonstrate synergistic or superaddative effects can be used to treat ophthalmic disorders at concentrations low enough to avoid undesired side effects.
Assay for Proinflammatory Cytokine-Suppressing Activity
[0345] Compound dilution matrices were assayed for the suppression of IFNγ, IL-2, and TNFα, as described below. The results from these experiments are set forth in FIGS. 2A-2LL.
[0346] IFNγ
[0347] A 100 μL suspension of diluted human white blood cells contained within each well of a polystyrene 384-well plate (NalgeNunc) was stimulated to secrete IFNγ by treatment with a final concentration of 10 ng / mL phorbol 12-myristate 13-acetate (Sigma, P-1585) and 750 ng / mL ionomycin (S...
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