Genetic associations with human longevity at the APOE and ACE loci.

In an effort to dissect the genetic components of longevity, we have undertaken case-control studies of populations of centenarians (n = 338) and adults aged 20-70 years at several polymorphic candidate gene loci. Here we report results on two genes, chosen for their impact on cardiovascular risk, encoding apolipoprotein E (ApoE), angiotensin-converting enzyme (ACE). We find that the epsilon 4 allele of APOE, which promotes premature atherosclerosis, is significantly less frequent in centenarians than in controls (p < 0.001), while the frequency of the epsilon 2 allele, associated previously with type III and IV hyperlipidemia, is significantly increased (p < 0.01). A variant of ACE which predisposes to coronary heart disease is surprisingly more frequent in centenarians, with a significant increase of the homozygous genotype (p < 0.01). These associations provide examples of genetic influences on differential survival and may point to pleiotropic age-dependent effects on longevity.

Genomic analysis of the human B-lymphotropic virus (HBLV).

The human B-lymphotropic virus (HBLV) has a double-stranded DNA genome of greater than 110 kilobase pairs, which is consistent with its morphological classification as a herpesvirus. A 9000-base pair cloned probe of HBLV detected specific sequences in DNA and RNA of infected cells but did not hybridize to the genomic DNA of other human herpesviruses including the Epstein-Barr virus, human cytomegalovirus, herpes simplex type I, and varicella-zoster virus. Conversely, while probes obtained from each of the known human herpesvirus readily detected the homologous viral DNA, they did not hybridize to genomic HBLV DNA. This evidence, in addition to serological and morphological distinctions and the biological effects of this virus demonstrate that HBLV is a novel human herpesvirus.

Increased poly(ADP-ribose) polymerase activity in lymphoblastoid cell lines from centenarians.

Poly(ADP-ribosyl)ation is a posttranslational modification of nuclear proteins which is catalyzed by poly(ADP-ribose) polymerase and represents an immediate response of eukaryotic cells to oxidative and other types of DNA damage. Previously a strong correlation had been detected between maximal poly(ADP-ribose) polymerase activity in permeabilized mononuclear leukocytes of various mammalian species and species-specific life span. To study a possible relation between longevity and poly(ADP-ribosyl)ation in humans we measured maximal oligonucleotide-stimulated poly(ADP-ribose) polymerase activity in permeabilized, Epstein-Barr virus transformed lymphoblastoid cell lines from a French population of 49 centenarians and 51 controls aged 20-70 years. Maximal enzyme activity was significantly higher in centenarians than in controls [median of controls: 9035 cpm/10(6) cells (lower quartile: 6156; upper quartile: 11,410); median of centenarians: 10,380 cpm/10(6) cells (lower quartile: 7994; upper quartile: 12,991); P=0.031 by Mann-Whitney U test]. In a subset of 16 controls and 24 centenarians, cellular poly(ADP-ribose) polymerase content was determined by quantitative western blotting, thus allowing the calculation of specific enzyme activity. The latter was significantly higher in centenarians (P=0.006), the median value for centenarians being about 1.6-fold that of controls. Specific poly(ADP-ribose) polymerase activity was a more powerful parameter for differentiating between centenarians and controls than enzyme activity relative to cell number. In addition, in a genetic association study we analyzed 437 DNA samples (239 centenarians and 198 controls) by PCR amplification of a polymorphic dinucleotide repeat located in the promoter region of the poly(ADP-ribose) polymerase gene in an attempt to detect an association between this polymorphic marker and variability of enzyme activity or human longevity. However, this genetic analysis revealed no significant enrichment of any of the alleles or genotypes identified among centenarians or controls, but its power was limited by the relatively weak heterozygosity of this polymorphic marker in our population (51%). Viewed together with previous results on poly(ADP-ribose) polymerase activity in various mammalian species, the present data provide further evidence for the notion that longevity is associated with a high poly(ADP-ribosyl)ation capacity.

New polymorphisms in the human poly(ADP-ribose) polymerase-1 coding sequence: lack of association with longevity or with increased cellular poly(ADP-ribosyl)ation capacity.

Poly(ADP-ribose) polymerase-1 (PARP-1) encoded by the PARP-1 gene, is a ubiquitous and abundant DNA-binding protein involved in the cellular response to various genotoxic agents. In a previous study we showed that maximal oligonucleotide-stimulated poly(ADP-ribosyl)ation was significantly higher in permeabilised lymphoblastoid cell lines from a French population of centenarians compared with controls aged 20-70 years, supporting the notion that longevity is associated with a genetically determined, high poly(ADP-ribosyl)ation capacity. Here, we describe four new genetic polymorphisms, three of which represent silent nucleotide variants (C402T, T1011C, G1215A), and one of which leads to a valine762-to-alanine exchange (T2444C). We undertook an association study between two of these polymorphisms and human longevity or poly(ADP-ribosyl)ation capacity in permeabilised lymphoblastoid cells. By analysing 648 DNA samples from a French population (324 centenarians and 324 controls) by fluorescent-allele-specific PCR, we showed the absence of any significant enrichment of any of the genotypes in the study of centenarians versus controls. Furthermore, we studied genotype distributions from individuals who had previously been tested for poly(ADP-ribosyl)ation capacity. None of the genotype combinations at any polymorphic site studied could be related to a high or low level of poly(ADP-ribosyl)ation capacity. Together, these results strongly suggest that the longevity-related differences in the poly(ADP-ribosyl)ation capacity of human lymphoblastoid cell lines cannot be explained by genetic polymorphisms in the PARP-1 coding sequence and that other mechanisms have to be considered as potential regulators of specific poly(ADP-ribosyl)ation capacity.

Palpable lymph nodes in healthy newborns and infants.

We examined 548 healthy neonates and infants to document the frequency, size, and location of palpable lymph nodes. The subjects consisted of 214 neonates from birth to 4 weeks of age and 334 infants from 4 weeks to 1 year of age. All of the infants were asymptomatic and had been free of major or minor systemic or cutaneous infections in the past. Of the 214 neonates, 73 (34%) had palpable nodes at one or more sites. Of the 334 infants, 190 (57%) had palpable lymph nodes. Inguinal, cervical, and axillary lymph nodes can be palpable in neonates and infants. Supraclavicular nodes are not generally palpable. The commonest site of palpable nodes is the inguinal area in neonates and the cervical area in older infants. It would appear that the palpable nodes noted in the neonatal period do not disappear but persist. This knowledge is useful in determining when adenopathy may be abnormal.

Cosleeping and sleep problems in Hispanic-American urban young children.

To assess whether the traditional pediatric prohibition against cosleeping in the parental bed requires reconsideration for urban ethnic minorities, cosleeping and sleep problems were studied in a sample of Hispanic-American, east Harlem, New York City, children 6 to 48 months of age. The incidence of frequent all-night cosleeping was found to be 21%, significantly higher than the documented rate of 6% found in a representative sample of white middle-American urban children of the same age and sex. For occasional cosleeping, however, there were no significant ethnic differences, and frequent part-night cosleeping was significantly less common than noted in the white sample. There were greater ethnic differences for sharing the parental bedroom compared with cosleeping in the parental bed, approximately 80% for Hispanic-Americans vs 10% for the white population. Within the Hispanic-American group, frequent all-night cosleeping was significantly more common among single parents and those living in multiple households and less common among infants and later-born children in the family. Frequent all-night cosleeping was also significantly associated with sleep problems.

Evidence of sex-linked effects on the inheritance of human longevity: a population-based study in the Valserine valley (French Jura), 18-20th centuries.

A long-standing puzzle in gerontology is the sex dependence of human longevity and its inheritance. We have analysed the sex-linked pattern of inheritance of longevity from 643 nuclear families on the historical population register of a French valley. We have focused on mean conditional life expectancy at a minimum age of 50 years, thus, in the present study, longevity refers to late or post-reproductive survival. A comparison of parents' and offspring's longevity has shown the existence of a heritable component of late survival in this population. We have found that the heritable component was substantially larger for daughters compared to sons. Moreover, this result appeared to be specific to late survival, that is, when only post-reproductive mortality for parental and offspring generations is taken into account. The stronger resemblance of parents to their daughters was no longer observed when considering younger ages at death for the offspring. This observation explains the hitherto unaccountable diversity of data in previous studies.

CD28 expression in T cell aging and human longevity.

Functional decrements of the immune system have a major contribution to aging and age-related diseases. Here, we further characterize the decline in proportion of CD28-positive T cells previously identified in centenarians. Cohorts of 97 centenarians, 40 subjects aged 70-90 (ELD group), and 40 young adults (under age 40) were phenotyped for T cell surface expression of CD28, CD4, and CD8 antigens. The significant decline in T cells expressing CD28 (p < 10(-4) for comparisons between adults and either ELD or centenarians) affects preferentially the CD8+ subset of T cells. This decline accounts largely for the age-related diminution of T cell responsiveness to mitogenic signals. CD28 expression is modulated in T cell cultures in a growth-related fashion and this modulation is dampened in cultures from centenarians. We propose that the decrease in CD28 expression reflects a compensatory adaptation of the immune system during aging in the face of chronic stimulation.

Decline in CD28+ T cells in centenarians and in long-term T cell cultures: a possible cause for both in vivo and in vitro immunosenescence.

The dramatic decline in immune function with age, especially in T cell proliferative activity, has been documented extensively in experimental animal models and in clinical studies of the elderly. A similar proliferative decline is also seen in long-term T lymphocyte cultures used to study in vitro cellular senescence. We have compared the peripheral blood T lymphocytes of centenarians and younger controls for the cell surface expression of CD28, a costimulatory molecule that is required for optimal activation and proliferation following engagement of the T cell receptor. Our analysis shows a significant decrease (p < 0.001) in the percentage of T cells expressing CD28 in the elderly cohort, with values ranging from 44% to 90%, as compared to the mean control value of 91%. The decline in the percentage of CD28+ T cells correlates with a reduction in the CD4/CD8 ratio (r2 = 0.695, p < 0.0001). Concommitantly, experiments using an in vitro T cell culture system showed a progressive loss of CD28 expression with culture "age." The concordance of proliferative decline and loss of CD28 in the centenarians and in the in vitro cultures suggest that a Hayflick phenomenon may operate in vivo leading to immunosenescence.

Genetics of survival.

The fields of gerontology and genetics have merged, spawning novel lines of investigation and generating a wealth of new results in recent years. However, the lack of clarity and consistency in the basic definitions upon which the science of gerontology must rest has fostered a certain amount of enduring confusion. Among the unclear issues are the genetic components of life span and the distinction between "normal" and "pathologic" aging. At a time of massive world population aging, such issues have, beyond their scientific importance, a momentous social and economic impact. A simple axiomatic framework, consisting of three definitions and five axioms, is proposed that clarifies the aforementioned issues and reconciles disparate data in gerontology. Based on this framework, a new classification of genes involved in survival is proposed. Within the Compensatory Adaptation Theory of aging, apparent paradoxes are solved and problems in gerontology may be formulated anew.

Contribution of cohort studies in understanding HIV pathogenesis: introduction of the GRIV cohort and preliminary results.

In the present paper we review studies performed on HIV-infected patients cohorts in order to understand AIDS disease development. The interplay between diverse factors such as the HIV envelope proteins, cellular co-receptors, the immune response with chemokines and cytokines production define the viral tropism, cytopathicity and progression of HIV disease. We present the trends of the research particularly in the domain concerning host genetics. In this context, we describe the GRIV cohort of fast and slow/non-progressors, and its use for understanding basic features of the yet unknown HIV pathogenesis mechanisms.

Homeostasis of chemokines, interferon production and lymphocyte subsets: implications for AIDS pathogenesis.

Certain individuals with elevated levels of macrophage inflammatory protein (MIP)1 alpha, MIP1 beta and RANTES expression appear to be resistant to human immunodeficiency virus (HIV) infection. In this work, we demonstrate that chemokines production by peripheral blood mononuclear cells (PBMCs) are homeostatic parameters varying from one individual to another, and we define optimized experimental conditions to reproducibly assess these parameters. We also studied alpha- and gamma-interferons (IFN alpha and IFN gamma, respectively) which have been implicated in the pathogenesis of acquired immunodeficiency syndrome (AIDS). The kinetics of production of all these cytokines by fresh PBMCs were determined upon stimulation with phytohemagglutinin (PHA), staphylococcus enterotoxin b (SEB) and purified protein derivative (PPD). RANTES and MIP1 alpha are produced early in response to activation, followed by MIP1 beta, (alpha-interferon, gamma-interferon, alpha IFN, gamma-IFN alpha and IFN alpha and gamma. These results suggest that using our methodology, chemokines levels can be reliably determined, permitting the performance of accurate genetic studies using PBMCs from various cohorts (siblings or AIDS related cohorts).

Pediatricians' and psychologists' implicit personality theory: significance of sibling differences.

Pediatricians' and psychologists' implicit theories of how personality develops are compared to recent behavioral-genetic analyses based on twin and adoption studies. Results show that both practitioner groups--less so the psychologists--underestimate differences between children in the same family, over-estimate the influence of their shared environmental experiences, and underestimate those not shared. Findings mirror the bias of traditional personality research, pointing out the need to investigate environmental variables with differential impacts on children in the same family, e.g., sibling deidentification; one child extrovert, one introvert; one saint, one sinner.

Sibling deidentification in the clinic: devil vs. angel.

A four-member family structure consisting of two siblings contrasting in personality (sibling deidentification) and each identified with a different parent (split-parent identification) was recently reported (15). In well-functioning families, this tetrad or quadrangle is wide-spread in the first pair of siblings in the family and tends to be more common in same-sex pairs, suggesting that sibling deidentification is designed to mitigate the relatively intense sibling rivalry characteristic of these pairs and hence to maintain family harmony. In this collated case report, deidentification is found to follow the same pattern in 39 clinic first pairs. However, contrasting attributes are varied and nonevaluative in nonclinic pairs but mainly "good-bad" in clinic pairs, with polarization extreme. Results suggest that nonclinic siblings negotiate their identity (being) much as they negotiate about possessions (having) and that negotiations are blocked in clinic pairs, freezing mythic devil or angel identity. Intervention is directed at dislodging this block.

[Genetics of longevity]. / Génétiques de la longévité.

It is possible to use different studies to demonstrate the genetic component of a phenotype. From the beginning of the century, many authors have studied the possible genetic transmission of longevity. The study of genealogies of ascendants first, and then of descendants of elderly individuals shows that the age at death incorrelated in people belonging to the same family. Finally, the studies carried out on monozygot and dizygot twins have made it possible to estimate that this genetic component accounts for approximately 10% of the individual's lifespan. Research on congenic mice which differ only by the chromosomic region of the Major Histocompatibility Complex (MHC) implies that this particular region might have an effect on longevity. However, the study of several alleles of the MHC indicates a complex sex-dependent influence. Several other chromosomic regions are also implied. As far as human beings are concerned, several research teams have worked on the HLA region. Once again, the situation is still far from clear.

Prospects for the genetics of human longevity.

Longevity varies between and within species. The existence of species-specific limit to human life-span and its partial heritability indicate the existence of genetic factors that influence the ageing process. Insight into the nature of these genetic factors is provided by evolutionary studies, notably the disposable soma theory, which suggests a central role of energy metabolism in determining life-span. Energy is important in two ways. First, the disposable soma theory indicates that the optimum energy investment in cell maintenance and repair processes will be tuned through natural selection to provide adequate, but not excessive, protection against random molecular damages (e.g. to DNA, proteins). All that is required is that the organism remains in a sound condition through its natural expectation of life in the wild environment, where accidents are the predominant cause of mortality. Secondly, energy is implicated because of the intrinsic vulnerability of mitochondria to damage that may interfere with the normal supply of energy to the cell via the oxidative phosphorylation pathways. Oxidative phosphorylation produces ATP, and as a by-product also produces highly reactive oxygen radicals that can damage many cell structures, including the mitochondria themselves. Several lines of evidence link, on the one hand, oxidative damage to cell ageing, and on the other hand, energy-dependent antioxidant defences to the preservation of cellular homeostasis, and hence, longevity. Models of cellular ageing in vitro allow direct investigation of mechanisms, such as oxidative damage, that contribute to limiting human life-span. The genetic substratum of inter-individual differences in longevity may be unraveled by a two-pronged reverse genetics approach: sibling pair analysis applied to nonagenarian and centenarian siblings, combined with association studies of centenarians, may lead to the identification of genetic influences upon human longevity. These studies have become practicable thanks to recent progress in human genome mapping, especially to the development of microsatellite markers and the integration of genetic and physical maps.

[Daltonism and the genetics of aging]. / Le daltonisme et la génétique du vieillissement.

In order to test whether mutations giving rise to color vision deficiencies are more frequently inherited from older fathers, an exhaustive screening of births in the Namur region has allowed to isolate a sample of 225 descending sons of maternal grandfathers who were older than 45 years at their daughter's birth. The incidence of color vision defects was compared between this set of cases and three control groups totalling 959 boys from independent families. While these comparisons were not conclusive, we propose new hypotheses concerning the population dynamics of color vision deficiencies. Neomutations in X-linked pigment genes may be a marker of the overall genetic load borne by the X chromosome. Selection against such loaded X chromosomes may occur in the second generation, either in the course of embryogenesis, or during female gametogenesis. The future assessment of these novel hypotheses relies on the arbitration of molecular genetics.

Lack of association between human longevity and genetic polymorphisms in drug-metabolizing enzymes at the NAT2, GSTM1 and CYP2D6 loci.

In the present study, the possible role of genetic polymorphism of three drug-metabolizing enzymes, debrisoquine/sparteine hydroxylase (CYP2D6), glutathione S-transferase mu (GSTM1), and N-acetyltransferase (NAT2), as a putative genetic component of human longevity, was explored. A total of 817 DNA samples from a centenarian and a control (20-70 years) population was subjected to PCR-coupled RFLP methods. Subjects were genotyped for the CYP2D6*3 (A2637 deletion) and CYP2D6*4 (G1934A transition) alleles, for four mutations of NAT2 [namely, NAT2*5A (C481T), NAT2*6A (G590A), NAT2*7A (G857A), and NAT2*14A (G191A)], and for the presence or absence of GSTM1 gene deletion. No significant difference was found at these three loci between centenarian and control subjects with respect to allelic variant frequencies, genotype distributions or predicted phenotypes deduced from genotype combinations. By comparing the distribution of combined genotypes for the polymorphisms tested at the CYP2D6, NAT2, and GSTM1 loci, none of the predicted phenotypes concerning debrisoquine hydroxylase extensive-metabolizer or poor-metabolizer phenotypes, slow or fast N-acetylation capacities, and active or defective glutathione S-transferase, could be correlated with human longevity, alone or in combination.

A model for antagonistic pleiotropic gene action for mortality and advanced age.

Association or linkage studies involving control and long-lived populations provide information on genes that influence longevity. However, the relationship between allele-specific differences in survival and the genetic structure of aging cohorts remains unclear. We model a heterogeneous cohort comprising several genotypes differing in age-specific mortality. In its most general form, without any specific assumption regarding the shape of mortality curves, the model permits derivation of a fundamental property underlying abrupt age-related changes in the composition of a cohort. The model is applied to sex-specific survival curves taken from period life tables, and Gompertz-Makeham mortality coefficients are calculated for the French population. Then, adjustments are performed under Gompertz-Makeham mortality functions for three genotypes composing a heterogeneous cohort, under the constraint of fitting the resultant mortality to the real French population mortality obtained from life tables. Multimodal curves and divergence after the 8th decade appear as recurrent features of the frequency trajectories. Finally, a fit to data previously obtained at the angiotensin-converting-enzyme locus is realized, explaining what had seemed to be paradoxical results-namely, that the frequency of a genotype known as a cardiovascular risk factor was increased in centenarians. Our results help explain the well-documented departure from Gompertz-Makeham mortality kinetics at older ages. The implications of our model are discussed in the context of known genetic effects on human longevity and age-related pathologies. Since antagonistic pleiotropy between early and late survival emerges as a general rule, extrapolating the effects measured for a gene in a particular age class to other ages could be misleading.

HLA-DR alleles display sex-dependent effects on survival and discriminate between individual and familial longevity.

In an effort to reassess the contribution of HLA-DRB1 polymorphisms to inter-individual variations of human longevity, we have compared their genotypic distributions between longevous and adult control groups in the French population. The longevous groups included two independent cohorts totalling 533 centenarians, and 163 nonagenarian siblings. Allelic distributions were significantly different between controls and longevous groups. Three individual alleles were mostly responsible for these differences: DR7, DR11 and DR13. Multivariate logistic analyses were performed in order to sort out interactions between gender- and age-specific genetic effects. DR7 frequency was elevated in longevous men, in centenarians as well as nonagenarian siblings [OR = 1.72 (1.2-2.5)]. DR11's influence on longevity displayed a significant interaction with sex, with an increase in women from longevous sibships [OR = 2.03 (1.4-3.0)]. DR13's frequency was increased in centenarians of both genders [OR = 1.46 (1.2-1.75)]. These results are discussed in the context of other pathophysiological effects of the implicated alleles. Our data support the direct involvement of three HLA-DR alleles in survival at very old ages. Two allele-specific effects on longevity appear to depend on gender and one on familial status for aggregation of this trait. The latter is an original finding for humans.