Table II.

Anti-CD28 enhances IFN-γ production from PBMCs stimulated with varying doses of PHA independent of CD40L or IL-12a

IFN-γ (pg/ml)
PHA 1/100PHA 1/500PHA 1/1000
Experiment 1
Media alone7,7174,956263
CD40T10,5327,4852,584
Anti-CD2821,1279,1801,136
Anti-IL-12<2001,062<200
Anti-CD40L2,285<200<200
Anti-CD28/anti-IL-121,6632,8951,329
Anti-CD28/anti-CD40L9,5952,274598
Experiment 2
Media alone6,6441,040330
CD40T15,1569,8805,259
Anti-CD289,5811,1601,830
Anti-IL-122,6321,184<200
Anti-CD40L3,051<200<200
Anti-CD28/anti-IL-12NDb1,4921,655
Anti-CD28/anti-CD40LND<2001,007
  • a In the same experiments as in Table I, fresh PBMCs were obtained from normal donors and stimulated with PHA 1/100 (3 μg/ml), PHA 1/500 (0.6 μg/ml), and PHA 1/1000 (0.3 μg/ml). Anti-IL-12 Ab (10 μg/ml) and/or anti-CD40L Ab (10 μg/ml) were added to cultures in the presence or absence of anti-CD28 Ab (10 μg/ml). Addition of CD40T (2 μg/ml) to cultures is also shown for comparison. Cells (2.5 × 105/250 μl) were added in triplicate to 96-well plates, and supernatants were assayed at 4 days for IFN-γ content by ELISA. Values shown are reported in picograms per milliliter. The sensitivity of the assay was between 100 and 200 pg/ml. SEM was ≤10% for all triplicate values. Data are representative of six separate experiments.

  • b ND, not done.