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U Fagiolo

Publications and source records attributed to U Fagiolo.

At least 19 recordsLinked to original sources
PubMed1998

Growth properties and growth factor responsiveness in skin fibroblasts from centenarians.

Human fibroblast cultures, which have a finite replicative lifespan in vitro, are the most widely used model for the study of senescence at the cellular level. An inverse relationship between replicative capability and donor age has been reported in human fibroblast strains. We studied the growth capacity of fibroblast primary cultures derived from people whose lifespan was as closer as possible to the expected maximum human lifespan, i.e. people over one hundred. Our data suggest that outgrowth of fibroblasts from biopsies, growth kinetics at different population doubling levels, capability to respond to a classical mitogenic stimulus (such as 20% serum) and a variety of growth factors, were remarkably similar in fibroblasts from centenarians and young controls. On the whole, our data challenge the tenet of a simple and strict relationship between in vivo aging and in vitro proliferative capability of human fibroblasts, at least at the individual level.

Adolescent↗
PubMed1998

Constitutive endothelial nitric oxide synthase (ecNOS) gene expression in human monocytes.

This study evaluates using a polymerase chain reaction (PCR)-based molecular biological approach to study the gene expression of the constitutive endothelial isoform of nitric oxide synthase (ecNOS) in human monocytes. When PCR was carried out with specific primers for ecNOS, 302 bp product was amplified, which sequence analysis determined as having 100% identity to the reported human ecNOS isoform gene sequence. The data presented here demonstrate a reliable technique for assessing ecNOS mRNA levels in circulating human monocytes. This then permits use of this easily available cell to monitor ecNOS levels and provides a means to investigate mechanisms involved in controlling NO synthase levels as well as NO synthesis.

Gene Expression↗
PubMed1997

Effect of rIL-2 treatment on anti-tetanus toxoid response in the elderly.

To explore the effects of interleukin-2 (IL-2) treatment in a vaccination protocol in the elderly, we administered low-dose rIL-2 to a group of aged subjects before primary tetanus toxoid immunization. A specific antibody response was detectable in the serum of 6/8 treated individuals after primary immunization, but in only 2/6 untreated controls; following antigenic boosting, specific antibody levels remained relatively unchanged in all the seroconverters. The data were confirmed by studying the ability to produce tetanus-specific antibodies in vitro, and by isoelectrofocusing analysis of serum anti-tetanus antibodies; this latter study showed a more restricted clonal response to the immunogen in untreated individuals. On the other hand, the study of the in vitro proliferative response to tetanus toxoid did not evidence clear differences between the two groups. On the whole, these data seem to indicate that a short-term rIL-2 treatment is able to potentiate the antibody response to tetanus toxoid, and may be a useful tool to improve humoral responses to vaccines in aged subjects.

Aged↗
PubMed1996

c-fos/c-jun expression and AP-1 activation in skin fibroblasts from centenarians.

In vitro replicative senescence is characterized by an irreversible growth arrest due to the inability of the cell to induce some key regulators of cell cycle progression, such as c-fos and AP-1, in response to mitogenic stimuli. In vitro replicative senescence and in vivo aging have been assumed to be two related phenomena, likely controlled by overlapping or interacting genes. As a corollary, fibroblasts from centenarians, which have undergone a long process of senescence in vivo should have very limited proliferative capability. On the contrary, in a previous work we found that fibroblasts from centenarians exhibited the same capacity to respond to different mitogenic stimuli as fibroblasts from young donors. Here we provide evidences that the well preserved proliferative response is likely due to the fact that some pivotal regulators- c-fos, c-jun and AP-1-are still fully inducible, despite a long process of in vivo senescence. Our data therefore suggest that in vivo and in vitro aging are separate phenomena whose possible relationships, if any, have to be ascertained very carefully.

Adolescent↗
PubMed1996

CD45 isoforms expression on CD4+ and CD8+ T cells throughout life, from newborns to centenarians: implications for T cell memory.

CD4+ and CD8+ peripheral blood T lymphocytes show mutually exclusive expression of CD45RA or CD45R0, two isoforms of the common leukocyte antigen that seem to recognize so-called virgin/unprimed and memory/activated T cells. The expression of these isoforms has been studied by three colour cytofluorimetric analysis on CD4+ or CD8+ peripheral blood CD3+ cells from 22 healthy centenarians, analyzed in a context of 202 healthy donors 0-110 years old. An age-related unbalance of virgin and memory cells was found between CD4+ and CD8+ subsets. As expected, at birgh 95-99% of the CD3+ lymphocytes expressed the CD45RA isoform. A rapid increase of CD45R0+ cells was observed in the first 2-3 decades of life, this phenomenon being much more pronounced on CD4+ cells. Subsequently, the increase of the 'memory' compartment was much less rapid, so that in centenarians a consistent reservoire of CD45RA+ among CD4+ cells was still present (about 20%). In these exceptional individuals the percentage of CD45RA+ cells among CD8+ T lymphocytes was even higher (about 50%), and only slightly lower than that of young donors (about 55-60%). Thus, the main changes occurred at a different rate in CD4+ (about 20%). In these exceptional individuals the percentage of CD45RA+ cells among CD8+ T lymphocytes was even higher (about 50%), and only slightly lower than that of young donors (about 55-60%). Thus, the main changes occurred at a different rate in CD4+ and in CD8+ T cells, at an age of between 0 and 30 years, when the thymus is still functionally active. Interestingly, no difference in the usage of CD45 isoforms was observed within T cells bearing four different V beta-T cell receptor (TCR). The significance of this age-related unbalance is unknown. However, the presence of a great number of CD45RA+ T lymphocytes within the CD4+ and the CD8+ T cell subsets even in the peripheral blood of centenarians poses the problem of their origin (thymus? extrathymic sites?), of their functional role and of their lifespan. Moreover, the data on centenarians suggest that they may represent a very selected population where a slowing of immunosenescence occurs.

Adolescent↗
PubMed1996

A shift to Th0 cytokine production by CD4+ cells in human longevity: studies on two healthy centenarians.

Centenarians, particularly healthy centenarians, constitute the example of successful aging and the study of their immune status can help to define the endpoint of the changes occurring throughout life. We characterized T cell clones (TCC) of two healthy centenarians, studying their phenotypes and production of representative Th1 and Th2 cytokines (IFN-gamma and IL-4) and compared them with TCC obtained by three young normal subjects; in all 180TCC were analyzed. In young donors, 35TCC were CD4+, 56CD8+ and 2 were alpha beta +CD4-CD8- (double negative). In centenarians, we obtained 46CD4+TCC, 38CD8+, 2CD4+CD8+ (double positive) and 1 gamma delta + double negative. Of the young subjects' TCC, 71% produced IFN-gamma but no IL-4 (Th1 pattern) and this prevalence decreased to 39% in TCC from the centenarians. The number of clones showing the opposite Th2 pattern was similar in young and aged donors (3 out of 93TCC and 2 out of 87TCC, respectively). The intermediate profile of TCC producing both IL-4 and IFN-gamma (Th0) was found in 25.8% of clones from young people, but it almost doubled to 58.6% in centenarians. The analysis shows that the Th profiles of CD8+TCC is nearly superimposable in the two groups, whereas a major shift from a Th1 to a Th0 pattern is presented by CD4+TCC. The balance provided by a majority of CD4+TCC showing a Th0 pattern may ensure both humoral and cell-mediated defences. In CD8+TCC, however, a Th1 pattern still is present, possibly for efficient generation of cytotoxic responses. These findings should be extended by studying other centenarians and elderly subjects.

Aged↗
PubMed1996

An immunological and genetic study of patients with gestational diabetes mellitus.

The aim of the study was to evaluate the frequency of islet cell (ICA) and insulin (IAA) antibodies and of HLA antigen typing in a group of subjects diagnosed with gestational diabetes mellitus (GDM) in a screening-diagnostic program during pregnancy. ICA, complement-fixing (CF) ICA and other autoantibodies, absolute number and percentage of lymphocyte subpopulations, and HLA antigens were evaluated in 68 women with GDM and compared with those of matched controls. ICA were found in 2 (2.9%) and IAA in 1 (1.5%). Both ICA-positive women had CF-ICA; one of them was receiving insulin therapy. while the other was on a special diet. No correlations were found between ICA and IAA, nor between IAA and insulin treatment. As far as lymphocyte subsets were concerned, we found a significant increase in the absolute number of total and activated (CD3+HLA-DR+) T lymphocytes and a significant increase in the absolute number and percentage of suppressor/cytotoxic T lymphocytes (CD8) and NK lymphocytes (CD57) in GDM patients compared with normal pregnant controls. Concerning frequency for HLA A, B, C, DR antigens in the GDM population, only Cw7 was found to be significantly increased and A10 significantly decreased in comparison with controls. Our study suggests that GDM is a heterogeneous disorder in which few patients present with the immunologic and genetic markers of type 1 diabetes.

Adult↗
PubMed1996

Blotting patterns of IgG anti-(U1)RNP antibodies in mixed connective tissue disease.

Serum reactivities towards individual U1 snRNP proteins were determined by immunoblotting in 32 patients with mixed connective tissue disease (MCTD). Time persistence of immunoblot profiles and clinical significance of anti-(U1)RNP antibody specificities were also investigated. IgG anti-(U1)RNP antibodies were found in the sera of 29 out of 32 patients (90.6%): 21 (65.6%) reacted with the 70-kD protein, 25 (78.1%) with A, 23 (71.9%) with C and 20 (62.5%) with B/B' proteins. None were reactive with the Sm-D peptide. Seventy kilodalton antibody specificity was strongly associated with a higher antinuclear antibody titre (> 160) and slightly associated with disease activity; anti-B/B' specificity was associated with lymphadenopathy. Anti-A, -C and -B/B' antibodies were negatively associated with systemic lupus erythematosus (SLE) skin rashes. Two types of anti-(U1)RNP blotting patterns were selected: "full spectrum" (53.1% of cases) and a "partially/no reactive" one (46.9%). Such patterns were unchanged over time in 14 out of 16 cases prospectively examined (87.5%), while the pattern shifted from "full spectrum" to "partially/no reactive" in 2 cases (12.5%): in 1 after a prolonged clinical remission (> or = 4 years) and in the other following immunosuppressive therapy. The anti-(U1)RNP antibody immunoblot profile in MCTD patients consisted of various reactivities and remained unchanged over time in most cases. Antibody reactivity against the 70-kD protein represented the major U1 snRNP specificity. The various anti-(U1)RNP specific reactivities demonstrated poor clinical significance within MCTD. Thus, MCTD seems to be characterized by a longstanding serological heterogeneity whose reactivities do not apparently correspond to distinct features within the broad clinical spectrum of MCTD.

Adult↗
PubMed1995

Thyroid autoimmunity and ageing.

Ageing is associated with the appearance of several serum autoantibodies, including thyroid autoantibodies. The biological and clinical significance of this phenomenon is still unknown, since, with the exception of primary myxedema, the prevalence of clinically overt thyroid autoimmune diseases is not increased in the elderly. The peculiar link between autoimmune thyroid failure and ageing is also underscored by the high prevalence of subclinical hypothyroidism in elderly subjects with positive serum thyroid autoantibodies, and could be the consequence of preferential age-dependent expression of destructive effector mechanisms and/or increased target gland susceptibility. Thyroid autoimmunity and subclinical hypothyroidism have also been implicated in the pathogenesis of other age-associated disorders, in particular coronary heart disease. Interestingly, recent data from our laboratories showed that thyroid autoantibodies are rare in healthy centenarians and in other highly selected aged populations, while they are frequently observed in unselected or hospitalized elderly. Taken together, these data suggest that thyroid autoimmune phenomena are not the consequence of the ageing process itself, but rather might be related to age-associated disease.

Aging↗
PubMed1995

Humoral response to influenza hemagglutinin: oligoclonal spectrotype and failure of thymopentin as immunoadjuvant.

Influenza remains a serious cause of illness and death among certain populations. Influenza vaccines in use at present are of limited effectiveness due to the high variability of the virus, and trials all over the world are in progress to enhance their immunogenicity. Conflicting results, in fact, have been reported about the immune response to influenza vaccination in diverse populations. In this paper we analyzed the antibody response to the hemagglutinin (HA) of the H3N2 A/Shangai 16/89 strain, which was included into the trivalent 1991-92 influenza vaccine, in four groups of subjects: 8 healthy young, 13 human immunodeficiency virus (HIV)-infected and 37 elderly healthy people, 9 of whom were treated with thymopentin (TP-5). Our results show levels of anti-HA IgG before vaccination in HIV-infected and elderly people significantly lower than those of normal young subjects. After vaccination, HIV-infected and elderly healthy people showed a significant increase of specific antibodies, whereas a failure in the specific response in normal young subjects was observed, thus differences among the groups were no longer present. Moreover, the spectrotypic analysis of antibody response, by isoelectric focusing and reverse blotting, showed oligoclonal but polymorphic pattern in the majority of subjects, irrespective of the group, and more frequently lack of expansion of the spectrotype after vaccination, thus demonstrating the lack of the recruitment of antigen-specific B cells. Finally, the treatment with TP-5 did not influence the outcome of the vaccination in the group of elderly people. These results further emphasize the limited immunogenicity of influenza vaccination and the inefficacy of TP-5 as immunoadjuvant, in this model of vaccination.

Adjuvants, Immunologic↗
PubMed1993

Age-related changes in human ceruloplasmin. Evidence for oxidative modifications.

Human plasma or serum from donors of age comprised between 15 and 95 years was analyzed for paramagnetic and total copper content, as well as for immunoreactive ceruloplasmin content and oxidase activity. All parameters were essentially unaltered, except the paramagnetic copper content, which increased 2-fold upon aging. A dramatic change of the electron paramagnetic resonance spectrum due to ceruloplasmin occurred in individuals over 65 years old and was associated with both an increase of the type 1 copper signal intensity and the appearance of new resonances of a type 2 copper species. Ceruloplasmin was isolated from either young or old donors. Spectroscopic analyses of the isolated proteins confirmed the tendency of type 1 copper to stay reduced in the "young" and oxidized in the "old" protein. The type 2 copper signal observed in most young ceruloplasmin samples was different from the species invariably present in the old protein. The magnetic parameters of the latter species were more consistent with a partially reduced trinuclear copper site. In vitro limited proteolysis resulted in identical fragmentation patterns and kinetics in both proteins. However, changes of the net electric charge were detected in the fragments of the protein isolated from aged individuals, which exhibited a carbonyl content of 0.6 mol of carbonyl/mol of protein. The same pattern of modifications, including a higher carbonyl content (0.65 versus 0.2 mol of carbonyl/mol of protein), could be reproduced by exposure of the young protein to the metal-catalyzed oxidation system iron/ascorbate. These results suggest that during aging ceruloplasmin is subjected to oxidative modifications which are likely to be the source of conformational changes around the copper sites leading to an intramolecular electron rearrangement among the various copper sites.

Adolescent↗
PubMed1993

Increased cytokine production in mononuclear cells of healthy elderly people.

The production of cytokines during aging, except interleukin (IL)-2, has been neglected in humans. We measured the in vitro production of IL-6, tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma and IL-1 beta by peripheral mononuclear cells from selected healthy young (mean age 26.8 years) and aged (mean age 80.2 years) subjects. Significant increases of IL-6, TNF-alpha and IL-1 beta levels were found in mitogen-stimulated cultures from aged donors, occurring at 24 to 72 h after stimulation. No significant differences were observed for IFN-gamma production. Proliferative capability of cells stimulated with PHA was not impaired in aged subjects. Since the amounts of all cytokines studied were similar in unstimulated cultures from young and aged subjects, and also serum levels of TNF-alpha did not differ, these data indicate that the cellular machinery for the production of these cytokines is well preserved in aging, and also that cells from old people are able to up-regulate their production in response to appropriate stimuli. The increases in cytokine synthesis were not dependent on changes in the number of monocytes, nor were they related to the significant rise of CD45RO+, and the concomitant decrease of CD45RA+, occurring in both CD4+ and CD8+ lymphocytes from aged subjects. The increased production of pro-inflammatory cytokines by stimulated mononuclear cells of healthy aged subjects may be relevant to several aspects of age-associated pathological events, including atherosclerosis, osteoporosis, fibrosis and dementia.

Adult↗
PubMed1993

Humoral and cellular immune response to influenza virus vaccination in aged humans.

Aging is characterized by an increased susceptibility to infectious diseases; influenza virus infection, which is easily managed by an intact immune system, represents a life-threatening disease in aged subjects. We studied 18 healthy aged subjects (> 65 years of age), vaccinated yearly with conventional anti-influenza vaccine, and 9 healthy young volunteers (mean age 26 years), without previous anti-influenza vaccination, who were vaccinated with the conventional trivalent 1990 anti-influenza preparation. Six out of the 18 aged individuals received a second boost of the same vaccine about 4 months later. In all subjects, we analyzed the humoral response to type A and B influenza viruses and the influenza type A virus-specific CTL generation. Among the elderly population with a single vaccination, 6 and 5 subjects seroconverted against type A and type B influenza virus respectively. Young subjects seroconverted in 5 cases against type A, and in 5 cases against type B influenza virus. Seroconversion took place after the second vaccination in only one subject, and the antibody production was type A specific. Influenza type A virus-specific CTL activity was significantly lower in aged subjects, compared with the values observed in the young volunteers (p = 0.017). The second vaccination partially restored this immunological impairment. These data clearly demonstrate that the elderly do not have the same ability as younger subjects to mount an antibody response, and generate influenza type A virus-specific CTL after conventional anti-influenza vaccination. Moreover, a double anti-influenza vaccination generates CTL activity levels comparable to young subjects, although it does not seem to substantially modify the antibody production.

Adult↗
PubMed1993

Quantitative and qualitative analysis of anti-tetanus toxoid antibody response in the elderly. Humoral immune response enhancement by thymostimulin.

In order to explore the humoral primary and secondary response to tetanus toxoid (TT), and to define the possible immunopotentiating effect of the thymic hormone thymostimulin, we studied 13 elderly people, selected according to the Senieur Eurage protocol, vaccinated against TT, an antigen never encountered before. Six of them were treated with thymostimulin before and during the immunization protocol. Specific anti-TT antibody level measurement and spectrotypic analysis were performed on the sera collected from the subjects at different times over the immunization protocol. In addition, spontaneous in vitro production of anti-TT antibodies as well as cutaneous delayed hypersensitivity reactions were also studied. Only one patient showed a detectable humoral immune response after the first immunization. After the booster, four of six thymostimulin-treated individuals, compared with only two of seven controls, showed in vivo anti-TT humoral response; at the same time, spontaneous anti-TT production was detected in peripheral blood mononuclear cells from five of six thymostimulin-treated individuals but only three of seven untreated controls. These differences were highly significant (p < 0.0001). In addition, only in thymostimulin-treated subjects were the levels of serum anti-TT antibodies 14, 21 and 28 days after the booster significantly (p < 0.05) higher than the baseline values. The spectrotypic analysis of anti-TT antibodies performed by isoelectric focusing and reverse blotting showed total agreement with the results from enzyme-linked immunosorbent assay.(ABSTRACT TRUNCATED AT 250 WORDS)

Adjuvants, Immunologic↗
PubMed1993

Complex alteration of thyroid function in healthy centenarians.

Several changes in thyroid function have been described in the elderly and largely attributed to concomitant nonthyroidal illness. The extent to which aging per se contributes to these changes remains to be elucidated, and scanty data are available in extremely old subjects. The present study was designed to focus on thyroid function during physiological aging, taking advantage of two groups of selected aged individuals: group A of healthy centenarians (n = 41; age range, 100-110 yr) and group B including healthy elderly subjects selected by the criteria of the EURAGE SENIEUR protocol (n = 33; age range, 65-80 yr). Control groups included 98 healthy normal adult subjects (group C; age range, 20-64 yr) and 52 patients with miscellaneous nonthyroidal illness (group D; age range, 28-82 yr). Our previous report of a low prevalence of thyroid autoantibodies in centenarians was confirmed and extended by the finding of a similar low autoantibody prevalence in the highly selected healthy elderly population of group B. Subclinical primary hypothyroidism was found in 3 (7.3%) centenarians, and their data were excluded from further statistical evaluation. No significant difference was found in the median serum free T4 levels of groups A-C. Median (and range) serum free T3 (FT3) was lower in centenarians [3.67 pmol/L (2.3-5.5)] than in group B [5.22 pmol/L (3.4-6.1)] and group C [5.38 pmol/L (2.9-8.4); P < 0.0001 vs. both groups]. Similarly, the median serum TSH level of centenarians [0.97 mU/L (< 0.09 to 2.28)] was lower than those in groups B [1.17 mU/L (0.53-2.74)] and C [1.7 mU/L (0.4-4.8); P < 0.0001 vs. both groups]; moreover, serum TSH was also significantly (P < 0.01) lower in group B than in group C. Both serum FT3 and TSH concentrations showed a significant inverse correlation (r = -0.634; P < 0.0001 and r = -0.377; P < 0.0001, respectively) with age. Median serum FT3 in centenarians was lower than that in group D patients [4.61 pmol/L (2.15-6.6); P < 0.0001]. In contrast, median serum rT3 in centenarians [0.40 nmol/L (0.20-0.77)], although higher than those in groups B [0.24 nmol/L (0.15-0.37); P < 0.0001] and C [0.22 nmol/L (0.05-0.46); P < 0.0001], was significantly lower than that in group D [0.60 nmol/L (0.13-2.08); P < 0.0001]. In conclusion, thyroid function appears to be well preserved until the eighth decade of life if healthy subjects are studied, whereas a reduction of serum FT3 is observed in extreme aging.(ABSTRACT TRUNCATED AT 400 WORDS)

Adult↗
PubMed1992

Thyroid and other organ-specific autoantibodies in healthy centenarians.

To investigate the prevalence of thyroid autoantibodies in very old subjects, we assayed sera from 34 healthy centenarians (7 men, 27 women; age range 100-108 years) for these antibodies. There was a clear age-dependent increase in prevalence of thyroid autoantibodies in sera from 549 control subjects aged 7-85 years, prevalence in 40 subjects aged 70-85 being significantly greater (p less than 0.001, chi 2) than that in 436 subjects aged less than 50. By contrast, prevalence of thyroid autoantibodies in centenarians was not significantly different from that in controls aged less than 50. Cytofluorimetric analysis of peripheral blood lymphocytes showed a striking age-dependent decrease in total and CD5+B cells (without changes in their ratio), which reached its nadir in centenarians. The age-dependent increase in prevalence of thyroid autoantibodies in the elderly is not seen after the ninth decade of life. What relation this characteristic has to derangement of circulating B cells is unknown.

Adolescent↗