As an HDAC inhibitor, n-butyrate alters the expression of a number of genes and their resulting Paclitaxel solubility dmso proteins. Among these proteins, the one best known to inhibit proliferation is the cyclin-dependent kinase

(cdk) inhibitor p21Cip1.10 p21Cip1 was up-regulated in T helper type 1 (Th1) cells anergized by exposure to n-butyrate.8 Recent studies in this model showed that p21Cip1-deficient CD4+ T cells were less sensitive than p21Cip1 wild-type CD4+ T cells to n-butyrate-induced anergy.11 p21Cip1 was not needed for the initial cell cycle blockade involved in anergy induction by HDAC inhibitors, but was required to maintain proliferative unresponsiveness when the anergic CD4+ T cells were restimulated with antigen. The mechanism by which p21Cip1 inhibited proliferation in the anergic CD4+ T cells was not defined, nor was it clear how p21Cip1, which is up-regulated under stimulatory as well as

tolerogenic conditions in CD4+ T cells, albeit with different kinetics, inhibits proliferation in the latter but not the former. p21Cip1 can inhibit cellular proliferation through at least three different mechanisms. As a cdk inhibitor, p21Cip1 selectively inhibits the enzymatic activity that is required for retinoblastoma protein phosphorylation and S phase entry. In accordance with this activity, overexpression of p21Cip1 has been shown to suppress cdk activity and cause G1 cell cycle arrest.12 PLX4032 in vitro p21Cip1 is also a potent inhibitor of the proliferating-cell nuclear antigen (PCNA), which is the processivity factor that functions as the sliding clamp on the DNA polymerase Rutecarpine delta, the principal replicative DNA polymerase. In resting T cells PCNA is low, whereas upon stimulation, PCNA expression increases 1000-fold during mid-G113 Inhibition of PCNA by p21Cip1 has been reported

to inhibit the cell cycle in both G1 and G2 phases in Jurkat T cells.14 The third mechanism by which p21Cip1 can block the cell cycle is through the inhibition of c-Jun N-terminal kniase (JNK). p21Cip1 has been shown to interact with JNK in vitro and to inhibit JNK activity in several cell types, including fibroblasts and T cells.15–17 JNK is a member of the mitogen-activated protein kinase (MAPK) signalling pathway that is activated by antigen stimulation in T cells. Triggering of the MAPK pathway in T cells normally leads to the activation of transcription factors such as activation protein 1 (AP-1), and to an associated increase in interleukin-2 (IL-2) transcription. However, in anergic T cells, defective IL-2 production has been linked to defects of JNK function, AP-1 activity and AP-1-dependent transactivation of IL-2 promoter,18–20 although the mechanisms for the defects observed are still unclear.