Local AngII can be produced by tissues and cells, including circulating cells, due to the presence of the complete enzymatic machinery required to synthesize and transform angiotensinogen (Jurewicz et al. 2007; Rasini et al. 2006). Moreover, AngII and its precursors can increase, in a dosage-dependent manner, the mitogen-stimulated T and NK cell proliferation thus playing a role in the induction of inflammatory re- sponse and oxidative stress (Jurewicz et al. 2007). In PMNs of patients with peripheral artery disease, in- creased levels of AT R have been demonstrated, sug- 1 gesting a role for AT R in the inflammatory processes 1 which occur in atherosclerosis (Marino et al. 2009). AT R stimulation of human neutrophils by AngII 1 − increases O and ROS production (El Bekay et al. 2 2003; Paragh et al. 2002) through increased cytosolic Ca++ levels, and such response is exacerbated in dia- betes (Inukai et al. 2005) and hypertension (Alba et al. 2008). Based on the above evidence, the results of the present paper might be taken to suggest that a reduced number of AT R on PMNs of subjects carrying the 1 minor rs275653 and rs422858 alleles translates into a reduced capability of such cells to generate ROS, thus creating anti-inflammatory state as to slow cell senes- cence. Further studies are needed to confirm this as- sumption and assess whether the variation observed in receptor expression only takes place in PMNs or is shared by other cell types.

Through AGTR1 exon sequencing of the Italian centenarians and younger controls, we discovered few novel variants, all but one in the non-coding regions. Interestingly, the only novel variant leading to amino acid substitution (c.893A>G, p.Asn298Ser) was found in a centenarian. This mutation is not in LD with promoter SNPs, and it is likely a private mutation. It occurs in the highly conserved NPxxY motif of most G-protein-coupled receptors, which is involved in the initiation of signal transduction following ligand bind- ing (Rosenbaum et al. 2009). Site-directed mutagenesis studies have previously shown that Asn298Ser replace- ment reduced the number of binding sites and impaired the receptor signalling in respect to the wild-type recep- tor, suggesting that the change found in our centenarian could affect AT R activity (Nikiforovich et al. 2006). 1 Further studies are needed to confirm this finding in humans. Our results indicate that genetic AGTR1 variants leading to reduced receptor expression may contribute to prolong human lifespan. These findings are of particular relevance as they disclose AT R as a 1 peculiar pharmacological target for the prevention and treatment of disabilities and diseases that occur with

Allelic variations in AGTR1 promoter, including the rs422858 major allele, have been associated with hy- pertension in Caucasian women (Kumar et al. 2005). Very recently, SNPs in AGTR1 promoter have been associated to the risk of hypertension also in Mexicans (Martinez-Rodriguez et al. 2011) and to salt sensitivity

Acknowledgments The generous support of Fondazione ART per la Ricerca sui Trapianti ONLUS (Milan, Italy) and of Fondazione Aiuti per la Ricerca sulle Malattie Rare, Bergamo, Italy, and of Istituto Edizioni ATLAS SpA, Bergamo, Italy is acknowledged. The authors thank the Istituto Italiano Edizioni ATLAS S.p.A for the grant for printing colour figures. The authors are indebted to Dr. Rossella Piras, recipient of a fellowship of Fondazione ART, for the invaluable help in gene sequencing and to Dr. Antonietta Chianca, Ph.D., for elrm analysis.

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