Journal:

Article Title: Interleukin 12 and B7-1 costimulatory molecule expressed by an adenovirus vector act synergistically to facilitate tumor regression

doi:

Figure Lengend Snippet: Expression kinetics and biological activity of IL-12 in AdIL12–B7-1-infected cells. The levels of IL-12 secretion following infection with AdIL12–B7-1 (moi 10) were determined by ELISA. (A) IL-12 expression in MRC5 (□), A549 (◊), and MT1A2 (▵). (B) Comparison of IL-12 levels in AdIL12–B7-1 (□), AdMEM12R (◊), and AdmIL12.1 (▵) infected primary FVBMT tumor cells. AdΔE1B7-1 (○) was used as a negative control. (C) Biological activity of IL-12 was tested by assaying the proliferation of Con A-activated mouse splenocytes in response to IL-12. FVB/n-mouse splenocytes (106 cells/ml) were incubated with supernatants from AdIL12–B7-1 and AdΔE1B7-1 infected MT1A2 (72 h p.i.) and primary FVBMT cells (48 h p.i.) for 24 h. After overnight incubation with [3H]thymidine, incorporation was measured by liquid scintillation counting. The IL-12 sample concentration was determined by comparison with a standard curve prepared using recombinant mIL-12. Five units of IL-12 are equivalent to 1 ng IL-12 protein. All samples were assayed in triplicate.

Article Snippet: The mIL-12 concentration for each sample was determined by comparison to a standard curve prepared using recombinant mIL-12 (gift of A. O’Garra, DNAX).

Techniques: Expressing, Activity Assay, Infection, Enzyme-linked Immunosorbent Assay, Negative Control, Incubation, Concentration Assay, Recombinant