there are no errors in the processed text.Polymorphisms Between Higher and Lower Age Groups  Distribution* Distribution* Genotype ,57 years (cid:2)57 years p Male Female p (cid:1)819 T/C C/C 26 (10.6) 22 (8.6) 25 (9.9) 23 (9.3) T/C 125 (51.0) 101 (39.6) 112 (44.5) 114 (45.9) T/T 94 (38.4) 132 (51.8) .0026y 115 (45.6) 111 (44.8) .8438y (cid:1)592 A/C A/A 47 (19.2) 56 (22.0) 49 (19.5) 54 (21.8) A/C 182 (74.4) 184 (72.1) 186 (73.8) 180 (72.6) C/C 16 (6.5) 15 (5.9) .0519y 17 (6.7) 15 (6.1) .6764y (cid:1)1082 A/G A/A 202 (82.5) 214(83.9) 218(86.3 )201(81 .0 ) A/G+G/G98(40 .0 )86(33 .7 )0 .2317x91(36 .1 )93(37 .5 )0 .7425x SNP single nucleotide polymorphism; IL interleukin; TNF tumor necrosis factor; TGF transforming growth factor; MMP matrix metalloproteinase.
In this study, we used custom-synthesized primers for PCR amplification and commercial Assays-on-Demand for IL-10 (cid:1)592 A/C. The reaction mixtures were loaded into 384-well plates and placed in an ABI Prism 7900HT Sequence Detection System for PCR amplification. We performed genotyping for eight SNPs, including IL-10 (cid:1)819 T/C, TNF-a (cid:1)1031 T/C, IL-1b (cid:1)511 C/T, IL-6 (cid:1)634 C/G, IL-18 (cid:1)607 A/C, TGF-b 869 C/T, MMP-1 (cid:1)1607 1G/2G, and MMP-3 (cid:1)1171 5A/6A. The relation of age and gender with the genotype distribution of each SNP was evaluated by chi-square test and multivariate logistic regression analysis. IL-10 (cid:1)819 T/C showed a significant association with age. The relation of IL-10 (cid:1)819 T/T with age was further investigated by dividing the population into four groups and comparing the frequency of IL-10(cid:1)819 T/T among them. We found that the frequency of IL-10(cid:1)819 T/T was increased in higher age groups compared to lower age groups. We also genotyped IL-10 (cid:1)1082 A/G and (cid:1)592 A/C to investigate the relation of a previously reported IL-10 haplotype with age, but there was no significant association with age.Single Nucleotide Polymorphisms and Age were studied in a Japanese population. The study showed a possible association of TNF-a (cid:1)1031 T/C with age and of IL-18 (cid:1)607 A/C and MMP-3 (cid:1)1171 5A/6A with gender, but multiple testing may cause a false-positive result. When evaluated by Bonferroni correction, only an association of IL-10 (cid:1)819 T/T with age attained statistical significance. However, the Bonferroni correction may overcorrect for the inflated false-positive rate and throw away valid information in the sample. A weak association should be regarded as a borderline statistical association at present and subjected to further investigations. The association of TNF-a (cid:1)1031 T/C with age and that of IL-18 (cid:1)607 A/C with gender are new findings, but the association of MMP-3 (cid:1)1171 5A/6A with gender was previously pointed out in coronary heart disease.

Control participants aged 25-53 years old showed an increased or decreased frequency of the genotype GCC/GCC or ACC/ATA, respectively, in an elderly group. However, no significant association of (cid:1)1082 A/G with longevity was observed in Finnish nonagenarians, an aged Irish population, or Sardinian centenarians. Pes and colleagues suggested that cytokine-longevity associations have a population-specific component affected by the population-specific gene pool as well as gene environment interactions.

The study group consisted of 163 people with mild chronic disorders except for malignancy. None of the SNPs showed a significant difference in genotype distribution between these 2 groups. Furthermore, there was no significant difference in genotype distribution and allele frequency on IL-10 (cid:1)819 T/C between our population and other Japanese populations previously reported, suggesting the absence of remarkable genetic heterogeneity in our population.

The genotype distribution of IL-10 (cid:1)1082 A/G is different from that of (cid:1)819 T/C or (cid:1)592 A/C in the Japanese population used in this study. This result was consistent with previous findings in a Japanese population, but differed from those in Caucasian populations, which revealed that these 3 polymorphisms were strongly linked, and only 3 of 8 possible haplotypes (GCC, ACC, ATA) segregated in Caucasian populations. In our population, IL-10 (cid:1)819 T/C was in perfect linkage disequilibrium with (cid:1)592 A/C, whereas it was not with (cid:1)1031 A/G. It was thus considered that there is little significance of this haplotype analysis in our population. The genotype distribution of these 3 polymorphisms was also quite different between our population and a Caucasian population. These findings clearly indicate a populational difference of the IL-10 promoter polymorphisms.

The effect of (cid:1)1082 A/G on IL-10 gene transcription or protein production seems to be controversial at present. Temple and colleagues reported the (cid:1)819 TT/(cid:1)592 AA haplotype as a determinant of high IL-10 transcription when peripheral blood mononuclear cells were stimulated with lipopolysaccharide or heat-killed Streptococcus pneumoniae, and IL-10 messenger RNA was measured by reverse transcriptase PCR. This finding might be supported by a recent association study showing that the IL-10 (cid:1)592 A/A genotype was associated with a decreased risk of grade III or IV acute graft-versus-host disease and death in remission in 933 recipients after hematopoietic cell transplantation. Further studies on different populations, and functional analyses of IL-10 (cid:1)819 and (cid:1)592 regions, are required to confirm the association of (cid:1)819 T/C and (cid:1)592 A/C with longevity.