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C D'Anna

Publications and source records attributed to C D'Anna.

15 recordsLinked to original sources
PubMed2001

A methodological strategy for PAH genotyping in populations with a marked molecular heterogeneity of hyperphenylalaninemia.

The elucidation of the molecular basis of hyperphenylalaninemia in various world populations (PKU Consortium Database: http://www.mcgill/ca/pahdb/) has revealed a remarkable molecular heterogeneity at the locus encoding for phenylalanine hydroxylase. As a consequence, genotyping of HPA patients has prompted the establishment of an impressive number of mutatIon detection protocols. In spite of the large variety of methods proposed so far, no comprehensive strategy has been yet developed for the detection of PAH gene mutations. Therefore, new approaches, combining the advantages of individual methods are required, especially in populations with a high number of PAH gene mutations. In this study, we propose the use of Reverse Dot Blot Analysis within a general mutation protocol to simplify the genotyping of hyperphenylalaninemics in the very heterogeneous population of Sicily (Italy).

DNA Mutational Analysis↗
PubMed2000

In vitro treatment with interleukin-2 normalizes type-1 cytokine production by lymphocytes from elderly.

The term immunosenescence is taken to mean the deterioration of immune function seen in elderly, which is manifested in increased susceptibility to infectious diseases, neoplasias, and autoimmune diseases. It is only recently that we have begun to understand the cellular and molecular changes involved. Of special interest in this regard are observations of a decline in synthesis of Type-1 cytokines which predisposes to diminished cell mediated immunity. We have evaluated the production of type 1 cytokines in old and young donors either in presence or in absence of recombinant interleukin-2 (rIL-2). Lymphocytes were stimulated with plastic bound anti-CD3 and after 48 h the supernatants were harvested and stored at -70 degrees C until assay. Type 1 cytokine, i.e. IL-12 and interferon-gamma (IFN-gamma) production by anti-CD3 stimulated lymphocytes from old subjects was significantly reduced when compared to that from young ones. This impaired production was reversed by adding rIL-2 in the culture medium. In previous studies on aged subjects, we have been able to demonstrate that in vitro treatment with rIL-2 completely restores proliferative responses and partially rescues the increased apoptosis of T cell cultures. Present and previous results suggest that rIL-2 completely restores Type 1 responses by overcoming the well known costimulation deficit of aged lymphocytes.

Adult↗
PubMed1999

Interleukin-5 production by mononuclear cells from aged individuals: implication for autoimmunity.

It is well known that in the elderly a deterioration of immune functions may occur. Particularly, stimulation of T cells from aged individuals leads to different kind and/or size of responses if compared with the responses obtained from T cells from young individuals. At the same time, an increase in prevalence of autoantibodies occurs in elderly. The altered production of certain cytokines might explain this paradox of decreased responsiveness to foreign antigens in the face of an increased response to self-antigens. We and others have suggested that this kind of immune response might depend on an age-associated impairment of Th-1 type function that selectively affects production of cytokines involved in the control of cellular responses. In contrast, Th-2 type function is seemingly not affected in elderly, as suggested by normal in vitro production of cytokines involved in humoral responses. To strengthen this hypothesis, in this study we have analysed the influence of age on the ability of mitogen-stimulated cultures of peripheral blood mononuclear cells from human beings to produce another Th-2 type cytokine, i.e. interleukin-5 (IL-5). IL-5 content of both 24- and 48-h stimulated cultures from old individuals was greater than that of young ones, although this difference attained significance only at 48 h. We suggest that the decreased production of Th-1 type cytokines in the presence of a normal or even increased production of Th-2 type cytokines might account for the pattern of immune response which may be observed in elderly, i.e. a normal or increased humoral response, including an autoimmune one, in the face of a low cell mediated immune response.

Adult↗
PubMed1999

Age-related changes in the expression of CD95 (APO1/FAS) on blood lymphocytes.

Aging is associated with alterations of the immune system, thought to be related to an increased susceptibility to infectious diseases, and possibly to cancer and autoimmunity in the elderly. In the present paper we report data obtained on freshly collected blood from 148 healthy subjects of different ages (from cord blood to 102 years old). The subjects were divided into seven age classes (cord blood, 3-11 years, 15-39 years, 41-60 years, 61-74 years, 75-84 years, 85-102 years) and their lymphocyte subsets and the expression of the apoptosis-related molecule CD95 were evaluated. In respect of lymphocyte subsets, the major differences were found in the cord-blood samples compared with the oldest old groups. In the cord-blood group, the absolute number of all the lymphocyte subsets was enhanced, but in the oldest group, an increase of CD16+ lymphocytes was observed, whereas CD19+ lymphocytes, which progressively decrease with age, continue to decrease further in the very old. The data show that the expression of CD95 increases until age 74 years, whereas in the oldest old it tends to decrease again. The trend of CD95 expression seems to be related to the change of expression of CD95 on CD4+ lymphocytes, because the CD8+/CD95+ population rose steadily throughout the entire age range. The evaluation of CD95+/CD45R0+ lymphocytes shows similar results to those observed analyzing CD95 on total lymphocytes. Furthermore, a constant increase of CD95+/CD28+ and a related decline of CD28+ lymphocytes was observed in all age groups. These data suggest that the expression of CD95 on the different subsets of lymphocytes can be considered a good marker for studies of immunosenescence, because it may be predictive of successful aging, and can partially explain the change in lymphocytes subsets in elderly.

Adolescent↗
PubMed1998

Interleukin-12 release by mitogen-stimulated mononuclear cells in the elderly.

Defects involving cellular expression of activation molecules, cell mediated immune response and natural killer (NK) activity are commonly observed in the elderly. Herein, data are reported on the evaluation of IL-12 production by old subjects. IL-12 is, actually, considered the key molecule for the induction of a T helper 1 (Th1) -type and NK response. IL-12 production from old subjects peripheral blood mononuclear cells (PBMNC) was evaluated using T-independent (bacterial lipopolysaccharide, LPS) or -dependent (phytoemagglutinin, PHA; immobilized anti-CD3 monoclonal antibodies, anti-CD3) mitogens. The IL-12 production after LPS stimulation was not reduced in cultures from old subjects when compared to that from young ones. On the contrary, IL-12 production by PHA or anti-CD3 stimulated PBMNC from old subjects was decreased. Furthermore, we have demonstrated a reduced CD40 and CD40 ligand (CD40L) expression on PBMNC from old subjects. This finding fits very well with the reduced cytokine production observed in the T-dependent stimulation systems, being the CD40-CD40L interaction mandatory for an efficient IL-12 production. All together, these results seem to suggest that defects in cell expression of activation molecules can affect the IL-12 secretion and in consequence other Th1-type cytokines.

Adult↗
PubMed1998

Apoptosis and ageing.

Stimulation of T cells from aged individuals leads to different kinds and/or size of responses if compared with the responses of T cells obtained from young individuals. In fact elderly is associated with a progressive decline of immune response besides an increasing incidence of autoimmune phenomena. These differences might be the result of modified cellular mechanisms controlling the immune system in the course of ageing. The apoptotic deletion of activated T cells has been proposed as the key mechanism to maintain T cell homeostasis, and in this respect CD95 (Fas antigen) seems to play a major role in this course of events. In this study we show that just collected lymphocytes from old subjects displayed an increased expression of the apoptosis molecule CD95. The expression of CD95 and the spontaneous apoptosis showed the same trend. In fact the percentage of apoptotic cells in blood collected from old subjects was enhanced too. The lymphocyte subpopulation analysis by flow cytometry did not show significant changes in T subset percentages between old and young subjects. Moreover mononuclear cells obtained from aged individuals underwent apoptosis in culture in response to a single stimulation with mitogen or anti-CD3, more than mononuclear cells from young controls. To gain insight into mechanisms of this increased apoptosis, experiments were performed to evaluate the behaviors of lymphocytes from old and young donors in respect of interleukin-2 (IL-2) rescue from apoptosis. Results show that IL-2 rescued only a little fraction of cells of old donors from apoptosis when activated by anti-CD3 and that this effect was not related to a different expression of CD95. Thus, during the course of ageing the different regulation of T cell homeostasis might be also explained by the modified proneness of lymphocytes to undergo apoptosis. The contemporaneous demonstration of a reduced Ca2+ influx in lymphoid cells of these subjects allows to suppose that multiple defects play a role in the pathogenesis of immunosenescence.

Adult↗
PubMed1998

Gluten stimulation induces an in vitro expansion of peripheral blood T gamma delta cells from HLA-DQ2-positive subjects of families of patients with celiac disease.

The intestinal gluten sensitivity formally known as celiac disease (CD) is characterized by an evident involvement of local immune response and it is associated with the expression of HLA-DQ2 allele. The major role in the disease seems to be played by the T lymphocyte population bearing gamma delta T cell receptor (T gamma delta cells) which are increased both in peripheral blood and intestinal mucosae of celiac patients. In this paper data on the effects of in vitro gluten stimulation on lymphocytes expressing the T gamma delta phenotype are reported. Gluten seems to be able to induce the expansion of the T gamma delta cell population both in CD patients and their HLA-DQ2-positive asymptomatic relatives, in spite of the absence of clinical evidence of the disease. In addition, the evaluation of gluten-induced cytokine production shows that interleukin-4 could be implied in the early phases of pathogenesis of CD.

Adolescent↗
PubMed1998

Biological basis of the HLA-B8,DR3-associated progression of acquired immune deficiency syndrome.

The factors influencing the evolution of human immunodeficiency virus (HIV) infection are not fully known, but the host genotype undoubtedly plays a role in determining the outcome of the disease by affecting the immune response to HIV. The role of the host human leukocyte antigen (HLA) genotype in the regulation of susceptibility to HIV infection and expression has been studied extensively in different major risk groups. Certain HLA alleles and haplotypes, being associated with aberrant immune responses independently from HIV infection, have been reported to facilitate the rapid progression of disorders related to HIV infection. Particularly, the association of rapid acquired immunodeficiency syndrome (AIDS) progression with genes from the HLA-B8,DR3 haplotype has been reported by different research groups. It is well known that this haplotype is associated in all Caucasian populations with a wide variety of diseases with autoimmune features and in healthy subjects with a number of immune system dysfunctions, as a reduced production of T helper (Th)1 type cytokine. HIV infection may act on this genetic background triggering immunopathogenetic mechanisms leading to AIDS with a dominant Th2 profile as a common feature.

Acquired Immunodeficiency Syndrome↗
PubMed1998

Hypothesis: interleukin-5 production impairment can be a key point in the pathogenesis of the MHC-linked selective IgA deficiency.

Selective IgA deficiency (IgA-D) is associated with the expression of some human leukocyte antigens (HLA) haplotypes and Major Hystocompatibility Complex (MHC) gene products have been suggested to be involved in the regulation of IgA synthesis. Recently, we have obtained evidences indicating that MHC influences the production of IgA and interleukin-5 (IL-5) both in humans and in mice. Lymphnode cells from pychril chloride (PC1) immunised BALB/c mice (bearing the H-2d haplotype) fail to produce IL-5 when stimulated in vitro with PC1 and this correlates with low antigen specific IgA production in vivo. In contrast using congenic BALB/k mice (bearing the H-2k haplotype) an high production both of IL-5 and of PC1-specific IgA is observed. Moreover, in vivo or in vitro administration of IL-5 to BALB/c mice was able to increase the production of antigen specific IgA. Similar evidences have been obtained by evaluation of the HLA influence on circulating immunoglobulin levels and interleukin production in normal HLA typed subjects. In fact HLA-B8, DR3 positive subjects show reduced level of serum IgA and their peripheral blood mononuclear cells stimulated with mitogen produce significantly reduced amounts of IL-5, IL-12, IL-2 and Interferon-gamma. We hypothesise that HLA-B8, DR3 associated IgA deficiency, known to be asymptomatic, can be due to a lack of subsequent signals, in particular of IL-5, involved in the late regulation of B cell differentiation. Preliminary evidences demonstrating that low amounts of human recombinant IL-5 are able to reconstitute IgA production by cells from HLA-B8, DR3 IgA-D subjects, seem to confirm this hypothesis.

Animals↗
PubMed1997

Modification of cytokine patterns in subjects bearing the HLA-B8,DR3 phenotype: implications for autoimmunity.

The factors influencing the pathogenesis of autoimmune disease are not fully known, but the host genotype undoubtedly plays a role in determining the outcome of these diseases. The role of the host's major histocompatibility complex (MHC) genotype in the regulation of susceptibility to autoimmune diseases has been extensively studied in different populations, and certain HLA (the human MHC) alleles and haplotypes have been reported to be associated with several autoimmune diseases. In particular, the association with genes from the HLA-B8,DR3 haplotype has been reported by different research groups. This haplotype is associated in all Caucasian populations with a wide variety of diseases with autoimmune features, and in healthy subjects it is associated with a number of immune system dysfunctions. Mainly, peripheral blood mononuclear cells from HLA-B8,DR3-positive and -negative individuals differ in their ability to produce interleukin (IL)-2, IL-5, IL-12 and interferon-gamma upon stimulation with the mitogen phytohaemoagglutinin (PHA), while producing similar amounts of IL4, IL-6 and IL-10. Furthermore, in HLA-B8,DR3-positive subjects tumor necrosis factor alpha secretion is increased both with and without PHA stimulation. Accurate control of the functional repertoire of an immune response is a critical parameter in the response to infections as well as in immunopathology. MHC control of the class of the immune response at the level of cytokine production is a sophisticated way in which this occurs. This control might be involved in adaptive immune responses to infections as well as in immunopathology.

Animals↗
PubMed1996

In vitro T cell activation in elderly individuals: failure in CD69 and CD71 expression.

A large number of T cell dysfunctions have been observed in the elderly. The most widely observed is the inability of these cells to proliferate at a level comparable to T cells from young individuals after stimulation by mitogens. To better characterize T cell impairment, we have focused on the in vitro T cell activation, analyzing by flow cytometry the activation molecules CD69 and CD71 on mitogen-stimulated lymphocytes from young and elderly subjects. The results show that the percentages of CD69+ and CD71 + T cells were significantly decreased in cultures from elderly subjects when compared to values obtained culturing cells from young individuals. The differences observed seem not due to differences in CD4 and CD8 rates in the "old' cells that underwent activation, since, following activation, the pattern of CD4 and CD8 phenotypes was the same in both groups of subjects. Signals from CD69 are relevant in controlling cytokine gene expression because its stimulation leads to interleukin-2 production and increases its receptor expression. The interaction of this cytokine with its cellular receptor is an essential requirement for T lymphocytes to express CD71 and to start proliferation. Thus, a key role in the age-associated impairment of T cell activation could be played by an ineffective modulation of CD69 expression suggesting a defect in the signal transduction pathway of the T cell receptor-CD3 complex in elderly.

Adult↗
PubMed1995

In vitro impairment of interleukin-5 production in HLA-B8, DR3-positive individuals implications for immunoglobulin A synthesis dysfunction.

Healthy subjects carrying the HLA-B8,DR3 haplotype may show a large number of immune dysfunctions. Concerning T-cell dysfunctions, the most intriguing is a defect of the early phases of T-cell activation, responsible for the impairment of in vitro mitogen-stimulated cytokine production. Regarding B-cell dysfunctions, one the most fascinating topics is the association between this haplotype and IgA deficiency in healthy blood donors. Accordingly, HLA-B8,DR3-positive healthy subjects show significantly lower values of serum IgA than HLA-B8,DR3-negative ones. Because IL-5 is a stimulating factor for the secretion of IgA by committed B cells, we have analyzed the in vitro mitogen-stimulated IL-5 production by MNCs from healthy HLA-B8,DR3-positive individuals to study whether they display an impaired production of IL-5. The results clearly demonstrate that MNCs from HLA-B8,DR3-positive individuals display significant reduction of IL-5 production, suggesting that IgA synthesis dysregulation observed in HLA-B8,DR3-positive subjects could be due to an impairment of IL-5 production.

Adult↗
PubMed1994

Viral antibody titers are influenced by HLA-DR2 phenotype.

Antibody serum levels against herpes simplex type 1 virus, cytomegalovirus, viral capsid antigens of Epstein-Barr virus, and rubella virus were evaluated in a sample of the Sicilian population. Results demonstrated that HLA-DR2-positive individuals showed a significant increase in antibody titers, when compared to HLA-DR2-negative individuals. These observations seem to be in contrast with the reported association of the HLA-DR2 phenotype with an ineffective immune response against several infectious pathogens. On the other hand, an increased humoral response to viral antigens need not be interpreted as a marker for effective control of virus infections. In fact, the response to virus infections is related to the T-cell-mediated immune response restricted by class-I- or class-II-encoded proteins. Thus, the above-mentioned HLA-DR2-related susceptibility to certain viral infections could be associated with a preferential induction of an increased (although ineffective) antibody synthesis against viral antigens.

Adult↗
PubMed1992

Risk factors and association with HBV infection in chronic C hepatitis.

The route of transmission in more than 50% of the patients with hepatitis C virus (HCV) infection is unknown. Only a minority of patients have had a previous blood transfusion; sporadic spread seems to be much more important, although the role of inapparent parenteral exposure is yet to be established. Aim of this study was to investigate if a relationship exists between history for exposure to known risk factors, concurrent HBV status and histological findings (presence of cirrhosis) in patients with chronic HCV liver disease. We studied 86 subjects with chronic HCV liver disease, subdivided according to their HBV status. Fifty four patients were anti-HBV negative; in the remaining 32 subjects, antibodies to HBV were found. Our data show that: 1) history for exposure to known risk factors is more likely to be present in patients with chronic HCV liver disease and concurrent positivity for antibodies to HBV than in anti-HCV positive patients without HBV antibodies (62.5% vs 38.9%); and 2) the incidence of liver cirrhosis is higher in anti-HCV positive patients with anti-HBV antibodies than in exclusively anti-HCV positive patients (56.2% vs 12.9%). We conclude that the association of history for exposure to known risk factors and anti-HBV positivity could be a marker of progression from mild to severe liver damage in patients with chronic HCV liver disease (i.e. in the absence of both identifiable risk factors and HBV antibodies, HCV infection could have a less severe clinical outcome). Therefore, in these patients a closer follow-up and earlier interferon therapy are probably needed.

Adolescent↗