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Fig. 1 | Journal for ImmunoTherapy of Cancer

Fig. 1

From: Characterization of the human T cell response to in vitro CD27 costimulation with varlilumab

Fig. 1

Cytokine induction, kinetics and TCR dependence of anti-CD27 costimulation of T cells by varlilumab or natural ligand (CD70). T cells were stimulated concurrently for 72 h with anti-CD3 (OKT3) and anti-CD27 (varlilumab; varli) antibodies. a Boxed whisker plot showing direct comparison of varlilumab stimulation of T cells with that of 4-1BB, OX40 and GITR-specific Abs [n = 6]. Statistical comparisons of IFNγ release by the different TNFR stimulations was performed using Welch Two Sample t-test; significance is shown as *p ≤ 0.05; **p ≤ 0.01. b Cell culture supernatants were tested for Th1/Th2 cytokines at the time points indicated by standard ELISA; representative data of 3 donor samples is shown; c T cells were stimulated for 72 h with OKT3 and irradiated 293-CD70 or 293-mock cell lines and assessed for IFNγ and IL-13. Additionally, CD70-dependent IFNγ release was assessed in the presence of a blocking Ab (BU.69) or control Ab (mIgG, left panel; n = 3) and IL-13 (right panel; n = 4); d T cells were first pre-activated for 46 h with anti-CD3 and further stimulated with varlilumab or hIgG or in combination with anti-CD3 for 24 h and tested for IFNγ (n = 4); NS, not significant (p = 0.09). e NFκB signaling by varlilumab-costimulated T cells for 72 h showing differential release of IFNγ in the presence of known pathway-specific inhibitors (top to bottom)- NFκB (Celastrol), ERK1/2 (PD98059), PKR (2-aminopurine), p38 MAPK (SB203580), IκBα (BAY11-7082), and JAK2 (AG490); bars represent mean ± SD (n = 4)

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