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In order to assess the training effect of a specific ligand upon a secondary stimulation, nontrained cells stimulated with LPS (or Pam3CSK4) were compared with trained cells restimulated with LPS (or Pam3CSK4).
To test the possibility that prior LPS or PS-F2 exposure would make macrophages refractory to subsequent PS-F2 stimulation, RAW264.7 cells were stimulated with LPS or PS-F2, then subjected to secondary stimulation with LPS or PS-F2 5 hours later.
Monocytes stimulated with Pam3CSK4 have been shown to be refractory to secondary stimulation with the same agonist [28].
Secondary stimulation with alloantigen showed similar results (Figure 3B).
Both labeled and unlabeled CTLs proliferated equally after the secondary stimulation.
Addition of anti-CD28 did not further increase mouse T cell infectivity (Figure 4C) The effect of secondary stimulation was initially observed in human cyclin T1 transgenic mice.
As opposed to phospho-IRF3, which could still be induced upon secondary stimulation with EBV, phospho-IRF7 progressively decreased under this condition.
For example, after secondary stimulation with proteolipid protein (PLP), labeled sensitized T-cells exhibited proliferation capacity similar to that of control, non-labeled cells [22].
In response to SEB secondary stimulation, T cell proliferation was transiently reduced in T cell cocultures with virus-matured MDDC, with the effect being greatest for IAV.
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We also reveal an action of TZDs to stimulate [H]-thymidine incorporation secondary to stimulation of uptake suggesting that other assays of proliferation are more suitable for studies with this class of drug.
Moreover, pre-stimulation of HEK.TPαS329A cells with U46619 completely desensitized [Ca2+]i mobilization and IP3 generation in response to secondary agonist stimulation (Fig. 5A C).
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