Papers by Domenico Frezza
Reumatismo, Dec 31, 2012
Objective. Several studies underline the relevance of the genetic background for the response to ... more Objective. Several studies underline the relevance of the genetic background for the response to therapy. We evaluated the relationship between the polymorphism of the HS1,2A enhancer, located in the 3' regulatory region of the heavy immunoglobulin chain (IgH), and the response to B cell depletion therapy (BCDT) with Rituximab (RTX). Methods. Fifty rheumatoid arthritis (RA) patients (42 women; disease duration 13.9±10.6 years) treated with RTX, not responsive to previous DMARDs and/or TNFα inhibitors therapies, and 220 healthy subjects were enrolled in the study. Patients were genotyped for HS1,2A enhancer polymorphism, as previously described. Disease activity was assessed every three months according to the European League Against Rheumatism's (EULAR) criteria. Results. All RA patients were seropositive for at least one of the tested autoantibodies: rheumatoid factor (FR IgA, FR IgM e FR IgG), anti-cyclic citrullinated peptides (anti-CCP IgA, anti-CCP IgM e anti-CCP IgG) and anti-vimentin antibodies. RA patients had an increased frequency of the allele*2 (60.0%) of the HS1,2A enhancer compared to healthy subjects (42.0%; OR(95%ICs): 2.07 (1.33-3.22)). Patients with a good EULAR response at 6 months follow-up visit had an increased frequency of genotype 2/2 (47.1%) compared to poorresponders RA patients (genotype 2/2: 18.2%, OR(95%ICs): 4.00 (1.09-14.68)). All the patients with a good EULAR response had the allele*2, thus showing a possible association with the allele in this population. Conclusions. The presence of allele*2 seems to be related to a good response to BCDT with RTX in seropositive RA patients, thus highlighting the role of the HS1,2A enhancer in B cell maturation and class-switch recombination.
Mutagenicity of industrial compounds: vinyl chloride, styrene and their possible metabolites
Mutation research, Apr 1, 1976
International Journal of Immunopathology and Pharmacology, 2009
Infectious and autoimmune pathogenic hypotheses of schizophrenia have been proposed, prompting se... more Infectious and autoimmune pathogenic hypotheses of schizophrenia have been proposed, prompting searches for antibodies against viruses or brain structures, and for altered levels of immunoglobulins. Previous experiments have shown that allele frequencies of the Ig heavy chain 3' enhancer HS1,2*A are associated with several autoimmune diseases, suggesting a possible correlation between HS1,2 alleles and Ig production. To test this, we analyzed levels of serum Igs and HS1,2*A genotypes in two independent cohorts, one of 88 schizophrenic inpatients (24 women) and a second of 133 healthy subjects (59 women). Both groups were similar in the frequency of individuals with altered serum concentration of Ig classes and IgG subclasses (schizophrenia panel-80%; controls-68%). With the possible exception of a stabilizing effect of olanzapine, no psychopharmacological drug consumed during the month prior to serum sampling in the schizophrenia group significantly affected Ig levels. In both patient and control cohorts, an increased frequency of the HS1,2 *2A allele corresponded to increased Ig plasma levels, while an increased frequency of the HS1,2*1A allele corresponded to decreased Ig plasma levels. EMSA analysis with nuclear extracts from human B cells showed that the transcription factor SP1 bound to the polymorphic region of both HS1,2*1A and HS1,2*2A while NF-κB bound only to the HS1,2*2A. We predict that differences in transcription factor binding sites in the two allelic variants of the 3' IgH enhancer HS1,2 may provide a mechanism by which differences in Ig expression are affected.
Annals of the Rheumatic Diseases, Oct 24, 2008
Objective: To investigate the role of the HS1,2 enhancer polymorphisms as a new candidate marker ... more Objective: To investigate the role of the HS1,2 enhancer polymorphisms as a new candidate marker for rheumatoid arthritis (RA) and to define the possible association with autoantibody positivity and clinical outcome. Methods: Genomic DNA was obtained from two cohorts of patients with RA (100 with early RA (ERA) and 114 with longstanding RA (LSRA)) and from 248 gendermatched controls from the same geographical area. Clinical and immunological characteristics were recorded for all the patients. Results: The percentage of the 2/2 genotype was higher in patients with ERA (27.0%), and in patients with LSRA (34.2%), than in controls (14.9%) (ERA: OR = 2.11 (95% CI 1.20 to 3.70) vs controls; LSRA: OR = 2.96 (95% CI 1.76 to 5.00) vs controls). A lower representation of allele *3 was present in patients with ERA (2.0%) than in controls (6.0%; OR = 0.32 (95% CI 0.11 to 0.91)). No significant associations were found between polymorphisms and autoantibodies positivity. Conclusion: The HS1,2A allele *2 associates with early and longstanding RA.
Nature Precedings, Dec 23, 2009
Association of HS1,2 alleles with autoimmune diseases.-Title : Association of autoimmune diseases... more Association of HS1,2 alleles with autoimmune diseases.-Title : Association of autoimmune diseases to allele 2 of the 3' immuno-globulines HS1.2 enhancer bearing an NF-κB binding site.
THU0239 Association of the 2/2 Genotype of the Enhancer Hs1,2A of the IG Heavy 3' Regulatory Region with Early Rheumatoid Arthritis
Annals of the Rheumatic Diseases, Jun 1, 2014
Background The allele*2 of the enhancer HS1,2A (IgH 3'RR-1) has been shown to have a binding ... more Background The allele*2 of the enhancer HS1,2A (IgH 3'RR-1) has been shown to have a binding site for NK-kB and to be associated to Rheumatoid Arthritis (RA), Systemic Lupus Erythematosus and Systemic Sclerosis. Objectives To study a polymorphism in the enhancer HS1,2A as a biomarker of disease activity and response to therapy in early RA. Methods 320 early RA (ERA) (age: 54.4±14.1 years; 76.3% female; 62.8% anti-CCP positive) were enrolled in the study. ERA patients fulfilled the 2010 American College of Rheumatology classification criteria for RA and were treated according to a tight control strategy. At baseline and every 3 months demographic and immunological data and the ACR/EULAR core data set were recorded. At each visit, clinical improvement and remission were evaluated according to DAS [1,2]. Patients were genotyped for HS1,2A enhancer polymorphism, as previously described [3]. Baseline IL-6 and BAFF plasma levels were evaluated by ELISA's methods. Results The analysis of the genotype's distribution of the HS1,2A enhancer showed a higher frequency of the 2/2 genotype in ERA patients (28.1%) compared to healthy subjects (14.9%, p<0.001). At diagnosis, ERA patients carrying the 2/2 genotype had a higher baseline inflammatory status (ESR: 49.6±30.5 mm/1st hr, CRP: 30.6±40.5 mg/l, IL6: 27.3±45.8 pg/ml) and higher disease activity (DAS28: 5.6±1.3, SDAI: 33.3±15.6) and disability (HAQ:1.3±0.8) compared to patients without the 22 genotype (ESR: 37.6±25.7 mm/1st hr, p=0.002; CRP: 20.1±27.3 mg/l, p=0.04; IL6: 20.6±38.5 pg/ml, p=0.04; DAS28: 5.3±1.3,p=0.05; SDAI: 29.2±14.1, p=0.06; HAQ:1.1±0.7, p=0.03). Similar autoantibody patterns were seen in the different genotypes. Fifty-three percent of ERA patients reached a good-EULAR response at 3 months follow-up visit (FU) and 24% were in sustained EULAR-remission at 6 months FU. The percentage of good-EULAR response at 3-months FU and sustained EULAR-remission at 6-month FU was lower in ERA patients carrying the 2/2 genotype compared with ERA patients without the 2/2 genotype [good-EULAR response: 36.2% vs 58.8%, OR (95%CIs): 0.40 (0.21-0.74); sustained EULAR-remission: 10.9% vs 27.9%, OR: 0.32 (0.13-0.79)]. Moreover, ERA patients carrying the 2/2 genotype were characterized by a higher percentage of subjects treated with TNF-blockers at 6 months FU compared with patients without the 2/2 genotype (20.3% vs 8.8%, p=0.02). Circulating B cells of subjects carrying the allele*2 showed a higher expression (fold change) of BAFF-R (7.6±3.7) and IL6 (4.1±6.4) than carriers of the 1/1 genotype (BAFF-R: 3.3±1.5, p=0.05; IL-6: 0.9±0.8, p=0.07). Conclusions The presence of the 2/2 genotype of the enhancer HS1,2A seems to identify a more active and aggressive disease in ERA patients poorly responsive to DMARDs. The presence of a binding site for NF-kB in the allele*2 (4), could explain the more aggressive phenotype. References van Gestel AM., Arthritis and Rheumatism, 1996. Prevoo ML, British Journal of Rheumatology, 1996. Tolusso B et al., Ann Rheum Dis 2009. Frezza D et al., Ann Rheum Dis 2012. Disclosure of Interest : None declared DOI 10.1136/annrheumdis-2014-eular.3591
Changes in virus-host genome relationship in two sublines of the same Burkitt’s lymphoma
Humana Press eBooks, 1989
The Burkitt lymphoma derived Namalwa cell line is the best studied case of EBV relationship with ... more The Burkitt lymphoma derived Namalwa cell line is the best studied case of EBV relationship with the human genome. The segments corresponding to those adjacent to the inserted viral genome have been cloned from non-infected human cell DNA (1).
Fems Microbiology Letters, Mar 1, 1999
The gene encoding for polynucleotide phosphorylase (pnp) of a new biovar of Staphylococcus aureus... more The gene encoding for polynucleotide phosphorylase (pnp) of a new biovar of Staphylococcus aureus subsp. aureus (NBSA) has been isolated from a genomic library of strain M280(0). The coding region consisted of a 1094-bp HindIII^HindIII DNA fragment encoding for a protein of 277 amino acids with a calculated molecular mass of 29.5 kDa. The nucleotide sequence of the structural gene, contained a continuous open reading frame of 836 bp, showed significant homology with the genes of bacterial polynucleotide phosphorylase from Bacillus subtilis (67.7% identity), from Haemophilus influenzae (62.47% identity), from Pseudomonas luminescens (61.6% identity), and from Escherichia coli (59.7% identity). DNA^DNA and DNA^colony slot-blot hybridizations demonstrated that the pnp gene, employed as a molecular probe, is specific for the identification of NBSA strains.
A marine host-mediated assay for the detection of mutagenic compounds in polluted sea waters
Mutation Research Letters, May 1, 1982
Marine water in heavily polluted areas may contain a variety of carcinogenic and mutagenic compou... more Marine water in heavily polluted areas may contain a variety of carcinogenic and mutagenic compounds (Phillips, 1978; Bend and James, 1978). Several methods have been developed to determine by chemical analysis the presence of carcinogens and mutagens in marine as well as fresh waters (Yevich and Borszez, 1976; Farley, 1976; Simmon et al., 1977; Lowe and Moore, 1978). Mussels, and many bivalves, can concentrate to a considerable extent substances present in the water (Bayne, 1976; Dunn and Stich, 1976; Mix, 1979). Extracts of mussels and other marine organisms spontaneously growing in polluted coastal areas of Great Britain and Florida were found by Parry et al. (1980) and Sparks et al. (1981), in experiments in vitro, to be mutagenic on yeast and bacteria. The Lagoon of Venice is heavily polluted, although to variable extents, by a number of chemical compounds, and data are available on the concentrations of several compounds in mussels and other marine organisms (Fossato et al., 1979). Battaglia and his group (1979) studied the modification of gene frequencies and the variation of heterozygosity level due to the effect of pollutants present in the Lagoon of Venice, on natural mussel populations and laboratory populations of the copepod Tisbe. In looking for a test more sensitive and with less manipulation than the techniques with extracts in vitro, we developed a host-mediated assay (h.m.a.). We used, as host organism, mussels grown in both clean and polluted areas, as well as clean mussels transferred for a period of time into polluted areas. We used 2 strains of yeast as genetic indicator microorganisms and then compared the results with those obtained by treating the same yeast strains, TA98 and TA100 of Salmonella typhimurium, with the corresponding mussel extracts. The use of h.m.a, for mutagenesis studies, where the organism under study is least manipulated during treatment, has been established as a reliable and sensitive method (Zeiger et al., 1972; Mailing, 1976; Pueyo et al., 1979).
Mutation research, Oct 1, 1979
Journal of Anatomy, May 1, 2008
After lentectomy, larval Xenopus laevis can regenerate a new lens by transdifferentiation of the ... more After lentectomy, larval Xenopus laevis can regenerate a new lens by transdifferentiation of the outer cornea and pericorneal epidermis (lentogenic area). This process is promoted by retinal factor(s) accumulated into the vitreous chamber. To understand the molecular basis of the lens-regenerating competence (i.e. the capacity to respond to the retinal factor forming a new lens) in the outer cornea and epidermis, we analysed the expression of otx2, pax6, sox3, pitx3, prox1, βB1-cry (genes all involved in lens development) by Real-time RT-PCR in the cornea and epidermis fragments dissected from donor larvae. The same fragments were also implanted into the vitreous chamber of host larvae to ascertain their lens-regenerating competence using specific anti-lens antibodies. The results demonstrate that there is a tight correlation between lens-regenerating competence and pax6 expression. In fact, (1) pax6 is the only one of the aforesaid genes to be expressed in the lentogenic area; (2) pax6 expression is absent in head epidermis outside the lentogenic area and in flank epidermis, both incapable of transdifferentiating into lens after implantation into the vitreous chamber; (3) in larvae that have undergone eye transplantation under the head or flank epidermis, pax6 re-expression was observed only in the head epidermis covering the transplanted eye. This is consistent with the fact that only the head epidermis reacquires the lens-regenerating competence after eye transplantation, forming a lens following implantation into the vitreous chamber; and (4) in larvae that have undergone removal of the eye, the epidermis covering the orbit maintained pax6 expression. This is consistent with the fact that after the eye enucleation the lentogenic area maintains the lens-regenerating competence, giving rise to a lens after implantation into the vitreous chamber. Moreover, we observed that misexpression of pax6 is sufficient to promote the acquisition of the lens-regenerating competence in flank epidermis. In fact, flank epidermis fragments dissected from pax6 RNA injected embryos could form lenses when implanted into the vitreous chamber. The data indicate for the first time that pax6 is a pivotal factor of lens-regenerating competence in the outer cornea and epidermis of larval X. laevis.
Mutation research, Jun 1, 1979
The strain SV3 of Salmonella typhimurium was used as the indicator bacterium in the intrasanguine... more The strain SV3 of Salmonella typhimurium was used as the indicator bacterium in the intrasanguineous host-mediated mutagenicity assay. Bacterial distribution and spontaneous mutation frequency were determined after intravenous injection of SV3 into CD~ male mice. Bacteria were cleared at an exponential rate from the blood stream and recovered mainly from the liver and in smaller quantities from the lungs and kidneys. No bactericidal effect was observed during incubation within the animal, and bacterial division occurred in the liver and probably in the kidneys. The significance of an increased mutation frequency of bacteria recovered from untreated animals is discussed. Mutation induction was measured in bacteria recovered from liver, lungs and kidneys of CD~ mice and CD rats treated with dimethylnitrosamine (DMN). The sensitivity of the intrasanguineous host-mediated technique was compared with the sensitivity of the assay in vitro with microsomal preparations from each tissue and host. Activation by isolated perfused liver and lungs from CD rats was included for comparison with the results from experiments in vivo and in vitro. The traditional host-mediated assay involves intraperitoneal injection of the indicator microorganism [12]. Modifications have been devised that bring the C.
Medical Sciences, Oct 5, 2018
An imbalance of bacterial quantity and quality of gut microbiota has been linked to several patho... more An imbalance of bacterial quantity and quality of gut microbiota has been linked to several pathologies. New strategies of microbiota manipulation have been developed such as fecal microbiota transplantation (FMT); the use of pre/probiotics; an appropriate diet; and phage therapy. The presence of bacteriophages has been largely underestimated and their presence is a relevant component for the microbiome equilibrium. As a promising treatment, phage therapy has been extensively used in Eastern Europe to reduce pathogenic bacteria and has arisen as a new method to modulate microbiota diversity. Phages have been selected and "trained" to infect a wide spectrum of bacteria or tailored to infect specific antibiotic resistant bacteria present in patients. The new development of genetically modified phages may be an efficient tool to treat the gut microbiota dysbiosis associated with different pathologies and increased production of bacterial metabolites and subsequently decrease systemic low-grade chronic inflammation associated with chronic diseases. Microbiota quality and mitochondria dynamics can be remodulated and manipulated by phages to restore the equilibrium and homeostasis of the system. Our aim is to highlight the great interest for phages not only to eliminate and control pathogenic bacterial infection but also in the near future to modulate the microbiota by adding new functions to selected bacteria species and rebalance the dynamic among phages and bacteria. The challenge for the medicine of tomorrow is to rethink and redesign strategies differently and far from our traditional thinking.
Isolamento e caratterizzazione di batteriofagi specifici per Pseudomonas syringae pv. actinidiae: un nuovo approccio per il trattamento della batteriosi del kiwi.Isolation and characterization of bacteriophages infecting Pseudomonas syringae pv. actinidiae: a new approach to the control of the ba...
Polymorphisms of the Ig heavy chain 3’ Regulatory Region associates with humoral Ig Homeostasis. (176.13)
Journal of Immunology, May 1, 2012
Aim. Incomplete maturation of immune system in the first years of life could affect different ind... more Aim. Incomplete maturation of immune system in the first years of life could affect different individual response to vaccination. To investigate the issue we studied a polymorphic genomic cis regulator of Ig heavy chain in response to vaccination with viral antigens. Method. Two cohorts of healthy subjects, one not yet 3 years and a second between 21 and 26 years old were vaccinated against H1N1, H2N3, B and HBV antigens respectively. From time zero the two cohorts were followed up for the total peripheral blood levels of Ig and for specific antibodies against the vaccine antigens. Subjects were genotyped for Ig heavy chain 3’ RR structural polymorphisms of enhancer hs1.2. Results. The serum specific antibodies in subjects Non responder, Responder and High responder to the vaccination is not associated with the levels of circulating IgM, IgG, IgA. However, the humoral concentration of Ig in the adult subjects correlates with hs1.2 allelic frequency as observed previously (p&lt;0.001). Children with low IgM concentration versus children with high IgM show a change of hs1.2 allele *1 frequency from 47% to 1.8% and for allele *2 from 43% to 82% (p&lt;10-9). The number of CD19 and CD27 cells were determined. Conclusions. The hs1.2 enhancer is associated with regulation of Ig levels. Children with allele *2 could have a possible advantage with wider repertoire of Ig. We suggest different controls for the production and mantainment of total and specific Ig amount with vaccination.
The functional VNTR of IGH enhancer HS1.2 associates with human longevity and interacts with TNFA promoter diplotype in a population of Central Italy
Gene, Nov 1, 2014
The dysregulation of both immune and inflammatory responses occurring with aging is believed to s... more The dysregulation of both immune and inflammatory responses occurring with aging is believed to substantially contribute to morbidity and mortality in humans. We have already reported the association of the functional Variable Number of Tandem Repeat (VNTR) at the Immunoglobulin heavy chain (IGH) enhancer HS1.2 with Immunoglobulin levels and with several autoimmune diseases. Herein we tested the association of the VNTR at the HS1.2 enhancer with human longevity, also evaluating the possible modulatory effect of TNFA promoter diplotype (rs361525/rs1800629). HS1.2 enhancer genotypes have been determined for 193 unrelated healthy individuals from Central Italy divided into two groups: Group 1 (18-84 yrs, mean age 56.8 ± 19.4) and Group 2 (85-100 yrs, mean age 93.0 ± 3.5). Homozygous subjects for 2 allele were significantly disadvantaged in reaching higher life-expectancy (OR=0.457, p=0.021). A significant interaction between TNFA promoter diplotype status, HS1.2 2/2 genotype and the two Groups was found (p=0.014). Of note, TNFA -308A allele seems to exert a protective effect in HS1.2 2/2 carriers. These results support the hypothesis of an important role of HS1.2 VNTR in the puzzle of the immune-system regulation, evidenced also by the potential interaction with TNFA. Moreover, the previous results showing the association of HS1.2 2 allele with inflammatory phenomena are consistent with the hypothesis that this allele is a detrimental factor in reaching advanced age.
Allelic frequencies of enhancer hs1.2 from Africa to Eurasia
Clinical Immunology, 2010
Asthma is characterized by chronic inflammation of airway mucosa and airway hyperresponsiveness (... more Asthma is characterized by chronic inflammation of airway mucosa and airway hyperresponsiveness (AHR). Asthmatic patients can respond to allergen provocation in
Birth Defects Research, 1984
Procarbazine (PCZ) was tested for its ability to induce mitotic gene conversions at the ade and t... more Procarbazine (PCZ) was tested for its ability to induce mitotic gene conversions at the ade and trp loci of Saccharomyces cerevisiae, strain D4. The influence of the following factors was examined: growth phase of the yeast cells (log vs stationary phase), pH of the treatment solution, and addition of mouse S9 fractions prepared from different organs. The drug was found more toxic and mutagenic at low doses (up to 25 mg/ml) for log phase cells, and scarcely toxic but highly mutagenic, even at high doses, for stationary phase cells. PCZ activity was reduced by acidic pH, and suppressed by S9 mix. Gene conversions were also analyzed in the intrasanguineous host-mediated assay performed in mice orally administered with PCZ. In such conditions PCZ was ineffective in stimulating mitotic gene conversions, probably owing to its inactivation in the acidic environment of the gastroenteric tract.
A Phage Therapy Model for the Prevention of <i>Pseudomonas syringae</i> pv. <i>actinidiae</i> Infection of Kiwifruit Plants
Plant Disease, Feb 1, 2023
Great efforts have been made with chemicals and pesticides to contain the spread of Pseudomonas s... more Great efforts have been made with chemicals and pesticides to contain the spread of Pseudomonas syringae pv. actinidiae (Psa) responsible for kiwifruit canker. Unfortunately, only partial results were obtained for this bacterial pandemic, and alternative remedies were proposed to avoid soil pollution and the onset of antibiotic resistance. Among these, phage therapy represents a possible tool with low environmental impact and high specificity. Several phages have been isolated and tested for the capacity to kill Psa in vitro, but experiments to verify their efficacy in vivo are still lacking. In the present study, we demonstrated that the phage φPSA2 (previously characterized) contains the spread of Psa inside plant tissue and reduces the symptoms of the disease. Our data are a strong indication for the efficiency of this phage and open the possibility of developing a phage therapy based on φPSA2 to counteract the bacterial canker of kiwifruit.
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Papers by Domenico Frezza