hile the second mode renders cells less sensitive to subsequent pulses of TNFa, if these pulses are separated by a short timespan. Later studies showed that the role of A20 goes beyond the 
control of NF-kB and that A20 is a general inhibitor in innate immune signaling; it protects cells from chronic inflammation, endotoxic shock and plays a role of tumor suppressor. In particular, A20 inhibits IRF3/IRF7 signaling. Similarly as for the NF-kB pathway, it acts upstream of the TBK1IKKE IKKc complex regulating negatively retinoic acid-inducible gene I protein , and potentially may act at the level of this complex by binding to IKKc. As said, the negative feedback loops involving IkBa and A20 lead to oscillatory responses. These oscillations Digitoxin site appear damped when analyzed at the population level, but single cell experiments by Nelson et al. on SK-N-AS cells and Tay et al. on 3T3 cells demonstrated that oscillations persist at least up to 10 hours. Discrepancy between population- and single cell-based observations can be explained by the progressing desynchronization of cells in the population, although the controversy about reconciling single cell and population data still exists. The major objection towards single cell experiments is that the additional gene copies coding for fluorescently tagged NF-kB may alter dynamics of the whole system. However, both experimental and modeling studies show that the number of NF-kB gene copies or its expression level influences only the amplitude but not the period of oscillations. Moreover, in our recent experiment, the expression of NF-kB remained practically unchanged due to the knockout of endogenous RelA, yet the oscillatory pattern was still clearly visible for 10 ng/ml TNFa dose. TNFa Autocrine and Paracrine Signaling TNFa affects growth, differentiation and function of cells of many types, and is a major mediator of inflammatory immune responses. It is considered as a key mediator of the septic shock syndrome induced by either LPS or bacterial superantigens. The potent activating abilities of TNFa are transmitted by 2 distinct cell-surface receptors: TNFR1 and TNFR2; the first one binds TNFa molecules with higher affinity and is considered responsible for the most of TNFa-induced signaling. It is established that binding of TNFa initiates proteinprotein interactions between TNFR1 and the TNFR-associated death domain protein. TRADD in turn recruits receptorinteracting protein and TRAF2 for NF-kB and survival signals. The TNFa autocrine and paracrine signaling arises since TNFa-inducible NF-kB serve itself as a primary transcription factor for TNFa. Over twenty years ago Collart et al. showed that TNFa promoter contains four kB motifs that can bind constitutive and inducible forms of NF-kB. Further analysis of kB motives in TNFa promoter revealed that two sites, kB2 and kB2a, play a primary role in TNFa regulation by NF-kB in response to 24900262 LPS stimulation in human monocytes. The autocrine regulation was observed in various cell lines and tissues: first, Wu et al. showed that TNFa functions as autocrine and paracrine growth factor in ovarian cancer. Coward et al. and Guergnon et 10408253 al. demonstrated that TNFa induces TNFa synthesis via NF-kB activation in human lung mast cells and B cells; Nadeau and Rivest found that in vivo TNFa injection induced TNFa mRNA expression in microglia and astrocytes, and later Kuno et al. showed that the activation of microglia by LPS is partially mediated by microglia-derived T