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Functional Display of IAb-p3K on the Surface of Insect Cells

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posted on 22.02.2013, 09:20 by Frances Crawford, Eric Huseby, Janice White, Philippa Marrack, John W Kappler

(A) Sf9 insect cells were infected with baculovirus encoding a membrane-bound form of IAb-p3K. After 3 d, the surface expression of IAb-p3K was detected with an anti-IAb mAb using flow cytometry.

(B) The genes for mouse ICAM (CD54) and B7.1 (CD80) were cloned into an insect cell expression plasmid as described in the Materials and Methods. The plasmids were used to cotransfect Sf9 cells, and a stable transfectant (Sf9-ICAM/B7.1) was cloned expressing both proteins detected with mAbs using flow cytometry.

(C) Either Sf9 (open bars) or Sf9-ICAM/B7.1 (closed bars) cells were infected with baculovirus expressing IAb-p3K. After 3 d, the infected insect cells were used as APCs to stimulate IL-2 production from B3K-06 and YAe-62. Uninfected cells were used as negative controls.


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