95 results on '"G. B. Ferrara"'
Search Results
2. A fuzzy approach to image analysis in HLA typing using oligonucleotide microarrays.
- Author
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G. B. Ferrara, L. Delfino, Francesco Masulli, Stefano Rovetta, and R. Sensi
- Published
- 2005
- Full Text
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3. Linkage of Chido and HL-A
- Author
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Judith A. Falk, Rodney Harris, E. Wolf, J.R. Batchelor, P. J. L. Cook, Janice Middleton, Sylvia D. Lawler, Marie C. Crookston, Flemming Kissmeyer-Nielsen, J. A. Sachs, Elizabeth B. Robson, Julia G. Bodmer, G. B. Ferrara, and H Festenstein
- Subjects
Male ,Canada ,Genotype ,Genetic Linkage ,Maximum likelihood ,Immunology ,Iceland ,Biology ,Biochemistry ,Genetic linkage ,Histocompatibility Antigens ,Genetics ,Humans ,Immunology and Allergy ,Antigens ,Israel ,Recombination, Genetic ,Linkage (software) ,Histocompatibility Testing ,Australia ,Chromosome Mapping ,Complement System Proteins ,General Medicine ,Cytotoxicity Tests, Immunologic ,Confidence interval ,Phenotype ,England ,Italy ,Austria ,Female ,Recombination Fraction - Abstract
Of 156 families who were HL-A typed and Chido-typed, 15 were found to be suitable for linkage analysis. It was calculated that the odds in favour of linkage of Chido and HL-A are 1,450,000:1. The maximum likelihood estimate of the Chido:1.HL-A recombination fraction is 2½% with 95% confidence limits, obtained graphically, of 0 and 12%; the estimate of the Chido:2.HL-A recombination fraction is 1½%, with limits of 0 and 9%. Among Chido negative subjects, the antigens HL-A12 and W5 occurred more frequently than in a control population.
- Published
- 2008
4. Alport syndrome: HLA association and kidney graft outcome
- Author
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P. Antonelli, Umberto Valente, M. Mossa, Sergio Barocci, G. Cannella, Gregorio Santori, Arcangelo Nocera, G. B. Ferrara, and S. Fiordoro
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Adult ,Male ,Alport Syndrome ,medicine.medical_specialty ,Linkage disequilibrium ,Humans ,Kidney Transplantation ,HLA ,Immunology ,Gene Expression ,Nephritis, Hereditary ,Human leukocyte antigen ,Biology ,Cohort Studies ,Type IV collagen ,Internal medicine ,Genetics ,medicine ,Alport syndrome ,Graft Survival ,HLA-DR Antigens ,medicine.disease ,Transplantation ,Phenotype ,Relative risk ,Cohort ,Female ,Nephritis ,HLA-DRB1 Chains - Abstract
Summary Alport syndrome (AS) is a genetic disease of type IV collagen involving non-homogeneous patterns of inheritance characterized clinically by the presence of progressive haematuric nephritis leading to end-stage renal disease (ESRD), hearing loss and/or ophthalmologic abnormalities. The aim of this study was to investigate, in a cohort of AS patients who had undergone a kidney graft (KG) or who were still on a waiting list for a KG, (a) whether there is a correlation between AS and HLA antigen expression, and (b) long-term graft outcome in transplant patients. The AS cohort was represented by 34 ESRD patients, of whom 25 received a KG and the remaining nine were still on a waiting list. AS transplant patients represented 2.78% of 899 first KGs performed at our centre (Transplantation Department at S. Martino Hospital, Genoa) between 1983 and 2002. Grafts were procured from cadaveric donors in 18 cases and from living, related donors in seven cases. All AS transplant patients had a post-transplant follow-up period of at least 12 months. Results showed that: (i) the frequency of the HLA-DRB1*16 antigen was significantly increased in the whole AS cohort as compared to 128 healthy subjects (HS) (corrected P-value 0.0026; relative risk 7.20) as well as to 232 non-AS ESRD patients on a waiting list for KG (corrected P-values 0.0156; relative risk 4.67); (ii) 5- and 10-year graft survivals in the AS transplant patients were 80 and 73%, respectively, and did not differ from those of a control group represented by 25 non-AS KG recipients matched for sex, age, number of HLA mismatches and immunosuppressive treatment. Increased frequency of HLA-DRB1*16 in AS patients may reflect a linkage disequilibrium with genes coding for collagen synthesis.
- Published
- 2004
5. HLA Class II DNA Typing in a Large Series of European Patients with Systemic Lupus Erythematosus
- Author
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Renzo Marcolongo, C. Papasteriades, Gian Domenico Sebastiani, Gabriella Morozzi, A. Jedryka-Goral, G. B. Ferrara, Carlo Carcassi, Enrique de Ramón Garrido, Antonio Fernández-Nebro, Josef S. Smolen, Mauro Galeazzi, Frédéric Houssiau, Ricard Cervera, Raffaella Scorza, J-C. Piette, G Porciello, and Giuseppe Passiu
- Subjects
Hla class ii ,HLA-D Antigens ,business.industry ,Autoantibody ,Case-control study ,Large series ,General Medicine ,chemistry.chemical_compound ,chemistry ,Case-Control Studies ,Immunology ,Humans ,Lupus Erythematosus, Systemic ,Medicine ,Genetic Predisposition to Disease ,Typing ,business ,DNA ,Autoantibodies - Published
- 2002
6. Identification of ricin A-chain HLA class II-restricted epitopes by human T-cell clones
- Author
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Deborah Castelletti, Marina Tommasi, Giulio Fracasso, Irene Lorenzetti, G. B. Ferrara, Giuseppe Tridente, Silvia Sartoris, Marco Colombatti, M. Pasti, and C. Pera
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T-Lymphocytes ,viruses ,T cell ,Molecular Sequence Data ,Immunology ,Priming (immunology) ,Human Immunology ,Biology ,Epitope ,chemistry.chemical_compound ,HLA-DR3 Antigen ,Immunotoxin ,medicine ,Humans ,Immunology and Allergy ,Amino Acid Sequence ,Allele ,Ribosome-inactivating protein ,Vaccination ,epitopes, immunotherapy, immunotoxins, ricin, T cell clones ,T cell clones ,HLA-DR Antigens ,T-Lymphocytes, Helper-Inducer ,epitopes ,biochemical phenomena, metabolism, and nutrition ,ricin ,Virology ,Peptide Fragments ,In vitro ,Clone Cells ,immunotoxins ,medicine.anatomical_structure ,Ricin ,chemistry ,immunotherapy ,Genes, T-Cell Receptor alpha ,HLA-DRB1 Chains - Abstract
Summary The identification of ricin toxin A-chain (RTA) epitopes and the molecular context in which they are recognized will allow strategies to be devised that prevent/suppress an anti-RTA immune response in patients treated with RTA-based immunotoxins. RTA-specific human T-cell lines and T-cell clones were produced by in vitro priming of PBMC. The T-cell clones used a limited set of Vβ chains (Vβ1, Vβ2 and Vβ8) to recognize RTA epitopes. The use of RTA deletion mutants demonstrated that T-cell lines and T-cell clones from three out of four donors responded to RTA epitopes within the domain D124-Q223, whereas one donor recognized the region I1-D124. The response to RTA peptides of T-cell lines and T-cell clones from two donors allowed the identification of immunogenic segments (D124-G140 and L161-T190) recognized in the context of different HLA-DRB1 alleles (HLA-DRB1*0801, and HLA-DRB1*11011 and B1*03011, respectively). The response to L161-T190 was investigated in greater detail. We found that the HLA-DRB1*03011 allele presents a minimal epitope represented by the sequence I175-Y183 of RTA, whereas the HLA-DRB1*11011 allele presents the minimal epitope M174-I184. RTA peptides and an I175A RTA point mutant allowed us to identify I175 as a crucial residue for the epitope(s) recognized by the two HLA-DRB1 alleles. Failure of T-cell clones to recognize ribosome inactivating proteins (RIPs) showing sequences similar but not identical to RTA further confirmed the role of I175 as a key residue for the epitope recognized in the context of HLA-DRB1*11011/03011 alleles.
- Published
- 2001
7. Cisplatin-vindesine-mitomycin (MVP) vs cisplatin-ifosfamide-vinorelbine (PIN) vs carboplatin-vinorelbine (CaN) in patients with advanced non-small-cell lung cancer (NSCLC): a FONICAP randomized phase II study
- Author
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M. Crippa, Noseda Ma, Elizabeth H. Baldini, Antilli A, Sunseri G, Carmelo Tibaldi, Cacciani Gc, Romano F, R. Rosso, Andrea Ardizzoni, Borghini U, M. Bandera, Galeasso G, Lepidini G, S. Barbera, L. Portalone, C. Lanfranco, Rinaldi M, M. Fioretti, Salvati F, M. Raimondi, M. C. Pennucci, Migliorino Mr, De Marinis F, and G. B. Ferrara
- Subjects
Cancer Research ,medicine.medical_specialty ,Chemotherapy ,Ifosfamide ,business.industry ,medicine.medical_treatment ,non-small cell lung cancer (NSCLC) ,Phases of clinical research ,medicine.disease ,Vinorelbine ,Gastroenterology ,Carboplatin ,Surgery ,chemistry.chemical_compound ,Oncology ,chemistry ,Internal medicine ,medicine ,Vindesine ,Lung cancer ,business ,medicine.drug - Abstract
In the present multicentre randomized phase II trial, the activity and toxicity of three platinum-based combination regimens for the treatment of advanced non-small-cell lung cancer (NSCLC) were evaluated. The three regimens were: MVP (mitomycin-C 6 mg m(-2) on day 1, vindesine 3 mg m(-2) on days 1 and 15, and cisplatin 80 mg m(-2) on day 1 every 28 days), PIN (cisplatin 80 mg m(-2) day 1, ifosfamide 3 g m(-2) day 1 and vinorelbine 25 mg m(-2) day 1 and 8 every 21 days) and CaN (carboplatin 350 mg m(-2) day 1 and vinorelbine 25 mg m(-2) days 1 and 8 every 28 days). A total of 140 chemotherapy-naive patients entered the study; 49 patients were treated with MVP, 48 with PIN and 43 with CaN. Sixty-seven per cent of the patients had stage IV disease. Response rates, calculated on an 'intention to treat' basis, were as follows: MVP, 14.3% (95% CI 5.94-27.2%); PIN, 16.7% (95% CI 7.4-30.2%); and CaN, 14% (95% CI 5.3-27.9%). The overall median survivals were 256, 269 and 243 days for patients treated with MVP, PIN and CaN respectively. Myelosuppression was the most frequent toxicity: grade 3-4 leucopenia was observed in 14.3%, 25% and 18.6% of patients treated with MVP, PIN and CaN respectively. This multicentre phase II randomized trial shows that MVP, PIN and CaN can be administered on an outpatient basis with acceptable toxicities. Unfortunately, the three regimens showed an activity significantly lower than that reported in previous single-institution phase II trials.
- Published
- 1998
8. Activation-related differences in HLA class I-bound peptides: presentation of an IL-1 receptor antagonist-derived peptide by activated, but not resting, CD4+ T lymphocytes
- Author
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G Frumento, A Corradi, G B Ferrara, and A Rubartelli
- Subjects
Immunology ,Immunology and Allergy - Abstract
We have compared by reverse phase HPLC the set of peptides eluted from HLA class I molecules in resting and activated CD4+ Jurkat cells. Two peptides were identified that are presented de novo upon activation. After sequencing, one of these peptides turned out to derive from IL-1R antagonist (IL-1Ra). In keeping with this observation, we found that activated, but not resting, Jurkat cells express IL-1Ra. These data indicate that activation of a CD4+ T cell line may result in presentation of peptides derived from proteins expressed de novo after activation. Since IL-1Ra was not known to be expressed by cells of the T lineage, we also investigated its pattern of expression in normal T lymphocytes. Reverse transcriptase-PCR analyses allowed us to demonstrate that IL-1Ra is expressed upon activation by normal CD4+ lymphocytes from peripheral blood and by thymocytes, but not by CD8+ T cells. Interestingly, of the two forms of IL-1Ra that have been detected in different cell lineages, the intracellular one and the secreted one, only the former is expressed by activated CD4+ T cells.
- Published
- 1997
9. Mutation in a splice-donor site of the APC gene in a family with polyposis and late age of colonic cancer death
- Author
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G. Del Porto, Paola Sala, Viviana Gismondi, L. De Benedetti, Paola Grammatico, L. Bertario, Silvano Presciuttini, Ray White, Anna Bafico, L. Varesco, Joanna Groden, Abdelhamid Heouaine, G. B. Ferrara, C. Rossetti, and Lisa Spirio
- Subjects
Adult ,Male ,Colorectal cancer ,Adenomatous polyposis coli ,RNA Splicing ,Molecular Sequence Data ,Biology ,Cell Line ,Exon ,APC gene ,Genetics ,medicine ,Humans ,Age of Onset ,Allele ,Child ,Frameshift Mutation ,Genetics (clinical) ,Aged ,Splice site mutation ,Base Sequence ,Autosomal dominant trait ,DNA ,Exons ,Middle Aged ,medicine.disease ,Exon skipping ,Pedigree ,Adenomatous Polyposis Coli ,biology.protein ,Female ,Age of onset - Abstract
Adenomatous polyposis coli (APC) is an autosomal dominant disease characterized by the development of hundreds of colorectal adenomatous polyps during the first decades of life. The expression of the disease varies, as the age of onset of colonic cancer and the severity of extracolonic manifestations often differ between affected families. An attenuated form of APC has also been described in which a small number of polyps and a later age of onset of colonic cancer is observed. Cloning of the APC gene has allowed disease-causing mutations in APC families to be identified. Here, we report a novel splice site mutation (a G to T transversion at position +5 of the splice donor site in intron 9) in the APC gene of affected individuals in an Italian family. Characterization of the transcription products from this mutant APC allele revealed that normal splicing was disrupted: a shorter mRNA was expressed in which exon 8 was connected directly to exon 10. This created a shift in the reading frame and the introduction of a stop codon at position 1358. In addition, some normal APC transcript was produced from the mutant allele in lymphoblastoid cells. A comparison of the clinical features of affected members of this family with four unrelated Italian APC kindreds, in which the same AAAAG deletion at position 3926 has been found, showed a significant difference in the onset of disease symptoms and in the age of death attributable to colorectal cancer. Inefficient exon skipping may be, at least in part, responsible for the delay in the development of the disease in the reported family.
- Published
- 1994
10. DNA TYPING OF HLA-DQ ALLELES BY GENE AMPLIFICATION OF DQA AND DQB VARIABLE EXONS: ANALYSIS OF DQA/DQB HAPLOTYPES
- Author
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G. B. Ferrara, C. Pera, A. Longo, G. Angelini, and Laura Delfino
- Subjects
musculoskeletal diseases ,endocrine system diseases ,Molecular Sequence Data ,Immunology ,Biology ,HLA-DQ alpha-Chains ,Linkage Disequilibrium ,law.invention ,Gene Frequency ,law ,HLA-DQ Antigens ,HLA-DQ ,Gene duplication ,Genetics ,HLA-DQ beta-Chains ,Humans ,Typing ,Allele ,Gene ,Alleles ,Polymerase chain reaction ,Base Sequence ,Histocompatibility Testing ,Haplotype ,Gene Amplification ,nutritional and metabolic diseases ,DNA ,Exons ,DNA Probes, HLA ,Transplantation ,Haplotypes ,Italy - Abstract
In the present report, we describe a DNA typing method that allows detection of all the polymorphic variants of DQA1 and DQB1 second exons. By the oligotyping procedure provided in this paper, we are able to identify 8 DQA1 and 13 DQB1 alleles and to type random individuals in any heterozygous combination. We provide the hybridization and washing temperatures for using either 32P labelled or non-radioactive probes. The discrimination power of this procedure, compared to serological and cellular techniques, is remarkable. Therefore, this typing method finds perfect application in transplantation immunology and it will be very helpful to optimize the matching of unrelated donors before BMT. It is apparent from our results that despite the linkage disequilibrium present between DQ and DR loci, a DR specificity may frequently be associated to different DQ haplotypes. This is the case for DR4, DR7, DR8, DR9, and DR13 specificities.
- Published
- 1992
11. Thenm23 gene maps to human chromosome band 17q22 and shows a restriction fragment length polymorphism withbg/II
- Author
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Generoso Bevilacqua, Ma Caligo, G. B. Ferrara, Mariano Rocchi, Dm Black, L Casarino, Ellen Solomon, Nardini, Paolo Simi, and L Varesco
- Subjects
Cancer Research ,Tumor suppressor gene ,Breast Neoplasms ,Hybrid Cells ,Biology ,Bacterial Proteins ,Gene mapping ,Cricetinae ,Genetics ,Animals ,Humans ,Genes, Tumor Suppressor ,Genetic Predisposition to Disease ,Kinase activity ,Deoxyribonucleases, Type II Site-Specific ,Gene ,Alleles ,BglII ,Monomeric GTP-Binding Proteins ,Chromosome Mapping ,Nucleic Acid Hybridization ,Proteins ,DNA ,NM23 Nucleoside Diphosphate Kinases ,Molecular biology ,Nucleoside-diphosphate kinase ,Blotting, Southern ,Chromosome Band ,Nucleoside-Diphosphate Kinase ,Restriction fragment length polymorphism ,Polymorphism, Restriction Fragment Length ,Chromosomes, Human, Pair 17 ,Transcription Factors - Abstract
The NM23-Hl gene is a putative tumor suppressor gene that may be important in the metastasic process. Recent genetic and immunological data indicate that the NM23-Hl gene encodes a protein with nucleoside diphosphate (NDP) kinase activity. The mapping of NM23-Hl by panels of rodent-human somatic cell hybrids and in situ hybridization showed that the gene is located in human chromosome band 17q22. A two-allele polymorphism with BglII was demonstrated.
- Published
- 1992
12. Analysis of Oligonucleotide Microarray Images Using a Fuzzy Sets Approach in HLA Typing
- Author
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R. Sensi, L. Delfino, Stefano Rovetta, G. B. Ferrara, and Francesco Masulli
- Subjects
business.industry ,Computer science ,Hybridization probe ,Fuzzy set ,Fuzzy control system ,Computational biology ,Human leukocyte antigen ,Bioinformatics ,Genome ,Transplantation ,ComputingMethodologies_PATTERNRECOGNITION ,business ,Genotyping ,Graphical user interface - Abstract
The Human Leukocyte Antigen (HLA) region is a part of genome which spans over 4 Mbases of DNA. The HLA system is strongly connected to immunological response and its compatibility between tissues is critical in transplantation. We have developed an application of oligonucleotide microarrays to HLA typing. In this paper we present a method based on a fuzzy system which interactively supports the user in analyzing the hybridization results, speeding-up the decision process moving from raw array data obtained from the scanner to their interpretation (genotyping). The two-level procedure starts with evaluation of spot activity, then it estimates probe hybridization levels from activity levels. The method is designed for being readily usable by the biologist, by adopting fuzzy linguistic variables which are familiar to the user and by featuring a standard and complete graphical interface.
- Published
- 2009
13. Amplification by the polymerase chain reaction of hypervariable regions of the human genome for evaluation of chimerism after bone marrow transplantation
- Author
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R. B. Wallace, Lawrence D. Petz, Priscilla Yam, D. Koyal, G. B. Ferrara, Richard E. Champlin, L. Ugozzoli, and S.J. Forman
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Genetics ,Oligonucleotide ,Immunology ,Cell Biology ,Hematology ,Biology ,Biochemistry ,Molecular biology ,law.invention ,Hypervariable region ,Transplantation ,Variable number tandem repeat ,Restriction enzyme ,law ,Human genome ,Restriction fragment length polymorphism ,Polymerase chain reaction - Abstract
We combined the polymerase chain reaction (PCR) with oligonucleotide hybridization as a novel and sensitive technique to evaluate posttransplant chimerism. Specific oligonucleotides for hybridization were synthesized homologous to tandemly repetitive core sequences of regions with a variable number of tandem repeats (VNTRs). Polymorphisms at such loci result from allelic differences in the number of repeats. Primers flanking the repeat region of each of the corresponding VNTRs were used for amplification. Recipient and donor pretransplant DNA and recipient posttransplant DNA were amplified. The resultant fragments were analyzed after gel electrophoresis either by hybridization in-gel or after Southern transfer. To confirm our findings, we also performed standard assays of restriction fragment length polymorphisms (RFLPs). Evaluation of 13 selected cases indicated mixed chimerism (4), complete chimerism (5), recurrence of leukemia (2), and endogenous repopulation of hematopoiesis (2) after marrow transplantation. Sensitivity of the method was determined by mixing various proportions of recipient and donor DNA; the limit of detection of the minor component in a mixture was 0.1%. PCR data correlated with RFLP data in all cases except two in which PCR proved more sensitive than RFLP. PCR amplification of VNTRs combined with oligonucleotide hybridization is a novel technique for documenting posttransplant chimerism and has advantages over RFLP analysis: high sensitivity, use of small amounts of DNA (250 ng), ease of preparation of DNA, elimination of need for restriction enzymes, and the ability to complete studies in 2 days.
- Published
- 1991
14. Combination Chemotherapy and Interferon α2b in the Treatment of Advanced Non-Small-Cell Lung Cancer
- Author
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C. Fortini, M. Crippa, G. Genovese, Editta Baldini, A. R. Cruciani, Andrea Ardizzoni, Antilli A, F. Salvati, G. Scagliotti, M. Rinaldi, E. Soresi, E. Gatti, C Pennucci, R. Tonachella, R. Rosso, and G. B. Ferrara
- Subjects
Cisplatin ,Cancer Research ,medicine.medical_specialty ,Chemotherapy ,Cyclophosphamide ,business.industry ,medicine.medical_treatment ,Alpha interferon ,Combination chemotherapy ,medicine.disease ,Gastroenterology ,Surgery ,Oncology ,Internal medicine ,medicine ,Lung cancer ,business ,Survival rate ,Interferon alfa ,medicine.drug - Abstract
Thirty-four patients with previously untreated advanced non-small-cell lung cancer were treated with a combination of polychemotherapy and recombinant interferon. Chemotherapy consisted of cyclophosphamide, 400 mg/m2, epidoxorubicin, 50 mg/m2, and cisplatin, 40 mg/m2 (CAP) i.v. on day 4; recombinant alpha 2b interferon (r alpha 2b IFN) was given i.m. daily at the dose of 3-5 MU from days 1 to 7. The treatment was repeated every 4 weeks. In the 32 eligible patients the overall response rate was 19.3% (95% C.L. 7.4-37.4%). Non-hematologic toxicity consisted formerly in flulike symptoms and fatigue complained of by 37.5% and 31.2% of patients, respectively, and vomiting reported in 68.7% of patients; grade III-IV myelotoxicity was observed in 12.5% of cases. In no case was the toxicity life threatening. The median overall actuarial survival and progression-free survival were 37 and 20 weeks, respectively. This study indicates that the combination of CAP chemotherapy and r alpha IFN is feasible and active in the treatment of advanced non-small-cell lung cancer.
- Published
- 1991
15. A fuzzy approach to image analysis in HLA typing using oligonucleotide microrarrays
- Author
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Stefano Rovetta, Francesco Masulli, L. Delfino, G. B. Ferrara, and R. Sensi
- Subjects
Logic ,business.industry ,Hybridization probe ,Human leukocyte antigen ,Computational biology ,Fuzzy control system ,Bioinformatics ,Fuzzy logic ,Genome ,Transplantation ,ComputingMethodologies_PATTERNRECOGNITION ,Artificial Intelligence ,business ,Genotyping ,Graphical user interface ,Mathematics - Abstract
The human leukocyte antigen (HLA) region is a part of genome which spans over 4 Mbases of DNA. The HLA system is strongly connected to immunological response and its compatibility between tissues is critical in transplantation. We have developed an application of oligonucleotide microarrays to HLA typing. In this paper, we present a method based on a fuzzy system which interactively supports the user in analyzing the hybridization results, speeding-up the decision process moving from raw array data obtained from the scanner to their interpretation (genotyping). The two-level procedure starts with evaluation of spot activity, then it estimates probe hybridization levels from activity levels. The method is designed for being readily usable by the biologist, by adopting fuzzy linguistic variables which are familiar to the user and by featuring a standard and complete graphical interface.
- Published
- 2005
16. HLA Typing Using a Fuzzy Approach
- Author
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Stefano Rovetta, G. B. Ferrara, and Francesco Masulli
- Subjects
Transplantation ,Decision support system ,Matching (graph theory) ,ComputingMethodologies_SIMULATIONANDMODELING ,Computer science ,Data mining ,Human leukocyte antigen ,computer.software_genre ,computer ,Fuzzy logic ,Histocompatibility - Abstract
The human leukocyte antigens (HLA) system consists of three regions in the human genome. In transplantation, the match between donor's and receiver's HLA is critical for histocompatibility. HLA typing problem consists in the donor's and receiver's HLA systems matching. We describe the image analysis module of a decision supporting system supporting the application of the oligonucleotide microarray technology to the HLA typing. The decision supporting system is based on a fuzzy modeling approach that allows the biologist to describe and classify the probe activations using its language and concepts in a natural way, and, at the same time, supports a robust interactive image filtering with the usage of a fuzzy basis functions network.
- Published
- 2004
17. HLA-C sequence based typing: nucleotide analysis from exon 1 through exon 8. Identification of a new allele: Cw*0718
- Author
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L, Delfino, A, Morabito, and G B, Ferrara
- Subjects
Molecular Sequence Data ,Humans ,HLA-C Antigens ,Sequence Analysis, DNA ,Polymerase Chain Reaction - Abstract
At present, 128 HLA-Cw alleles have been described. Twenty-four of 128 display critical polymorphisms in contributing to allele identification outside exons 2 and 3. As a matter of fact, complete resolution of Cw*030201, Cw*030202, Cw*0409N, Cw*0501, Cw*0503, Cw*070101, Cw*070102, Cw*070401, Cw*0706, Cw*0711, Cw*0718, Cw*120201, Cw*120202, Cw*150501, Cw*150502, Cw*1701, Cw*1702, Cw*1703, Cw*1801 and Cw*1802 alleles requires nucleotide analysis of exons 1, 4, 5, 6 and 7. Moreover, some alleles (Cw*04010101, Cw*04010102, Cw*07020101 and Cw*07020102) showing nucleotide differences outside the coding regions of HLA-C gene (intron 2) have been reported. High resolution sequence based typing (SBT) developed in this study involves two DNA amplifications and 12 direct sequencing reactions and allows the analysis of HLA-C polymorphisms from exon 1 through exon 8, including intron 2. This typing procedure identifies all 128 Cw alleles described so far. Nevertheless, a number of ambiguous heterozygous typing results may be expected, this being the major drawback of SBT methods. A total of 201 samples were HLA-C typed using SBT strategy here described. The sequence of exons 6, 7 and 8 of HLA-Cw*070102 allele was elucidated. A novel HLA-Cw*07 allele, Cw*0718, was identified in two samples. Cw*0718 differs from the Cw*070101 allele by a unique nucleotide position within exon 6, resulting in an amino acid substitution at codon 324 (Ala--Val) in the cytoplasmic region of the molecule.
- Published
- 2003
18. Identification of a new DRB3*02 allelic variant (DRB3*0209) by high-resolution sequence-based typing
- Author
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A, Morabito, C, Pera, A, Longo, L, Delfino, and G B, Ferrara
- Subjects
Male ,Polymorphism, Genetic ,Base Sequence ,Haplotypes ,Histocompatibility Testing ,Molecular Sequence Data ,Humans ,Female ,HLA-DR Antigens ,HLA-DRB3 Chains ,Polymerase Chain Reaction ,Sequence Alignment ,Alleles - Abstract
The HLA-DRB3/B4/B5 sequence-based typing method developed in this study in combination with PCR-SSP, enabled us to identify a new DRB3*02 allele, that was named as DRB3*0209 (GenBank accession number AF148518). This name has been officially assigned by the WHO Nomenclature Committee in May 1999. The new allele differs from DRB3*0207 by one substitution in codon 51 from AGG to ACG and another in codon 60 from TAC to TCC, resulting in aminoacid changes from Arg--Thr (codon 51) and from Tyr--Ser (codon 60). The DRB3*0209 allele was discovered in two related North Italian families. The fact that it was present in an hemizygous situation in three members of the paternal family and in one member of the secondary related family enabled us to isolate and sequence the new DRB3 allele without cloning, to identify its association with the DRB1 locus, and to generate an Epstein-Barr virus (EBV)-transformed cell line, now present in our ECBR (European Collection for Biomedical Research) Cell Line Bank.
- Published
- 2000
19. Melanomas and melanoma cell lines do not express HLA-G, and the expression cannot be induced by gammaIFN treatment
- Author
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G, Frumento, S, Franchello, G L, Palmisano, M R, Nicotra, P, Giacomini, Y W, Loke, D E, Geraghty, M, Maio, C, Manzo, P G, Natali, and G B, Ferrara
- Subjects
HLA-G Antigens ,Interferon-gamma ,HLA Antigens ,Blotting, Western ,Histocompatibility Antigens Class I ,Tumor Cells, Cultured ,Antibodies, Monoclonal ,Humans ,RNA, Messenger ,Flow Cytometry ,Melanoma - Abstract
HLA-G is an effective ligand of natural killer (NK) inhibitory receptors, HLA-G transcripts have been detected in several human tumors, and cytokines like gamma interferon (IFN) enable HLA-G molecules to be expressed. These findings are particularly upsetting in case of melanomas: IFN treatment is frequently included in melanoma therapeutic protocols, and downregulation of classical class I molecules occurs in nearly half of these tumors. Therefore, a melanoma cell downregulating classical class I and de novo expressing HLA-G, either constitutively or upon IFN treatment, is probably a stealthy target for the immune system, having inhibited both the cytotoxic T lymphocyte (CTL) and the NK activity. To elucidate this point we have investigated the expression of HLA-G molecules in 45 melanoma cell lines before and after gammaIFN treatment. Analysis was performed by immunofluorescence and flow cytometry, using the anti-HLA-G MoAbs 87G and G233, by Western blot, using the anti-HLA-G MEM/G1 MoAb and PAG1 antiserum, and by RT-PCR analysis. In addition, 8 melanoma tissues from patients free from therapy and 6 nevi were studied by immunohistochemistry using the 87G MoAb. No evidence was gathered of HLA-G expression, neither constitutive nor, in cell lines, after gammaIFN treatment. We therefore conclude that HLA-G expression is an uncommon event in melanomas, and that a therapy including IFNs cannot harm the patient by inducing the de novo expression of HLA-G molecules at least in its G1 isoform.
- Published
- 2000
20. Molecular characterization and applications of recombinant scFv antibodies to CD152 co-stimulatory molecule
- Author
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M P, Pistillo, P L, Tazzari, J H, Ellis, and G B, Ferrara
- Subjects
Intracellular Fluid ,Immunoconjugates ,Membrane Glycoproteins ,Recombinant Fusion Proteins ,T-Lymphocytes ,Immunoglobulin Variable Region ,Lymphocyte Activation ,Antigens, Differentiation ,Abatacept ,Mice ,Antigens, CD ,Antigens, Surface ,B7-1 Antigen ,Tumor Cells, Cultured ,Animals ,Epitopes, B-Lymphocyte ,Humans ,CTLA-4 Antigen ,B7-2 Antigen ,Immunoglobulin Fragments - Abstract
Recombinant human monoclonal antibodies against CD152 have been generated by selecting a synthetic phage scFv library with purified CD152-Ig fusion protein. Sixteen scFv fragments were isolated which specifically react with CD152 by enzyme-linked immunoabsorbent assay (ELISA) and Western blot resulting in their clustering into two groups recognizing different antigenic determinants. One group of scFvs (#3, #13, #40, #44, #47, #51, #57, #80 #83) recognized an epitope on CD152 dimer whereas another group (#15, #18, #31, #35, #54, #72, #81) recognized an epitope on both dimeric and monomeric CD152 molecule suggesting their possible use in understanding the subunit structure of CD152 which is still controversial. Sequencing of the VH genes revealed that all the scFvs belonged to the VH3 gene family but they were different in CDR3 length and composition. It was possible to correlate specific CDR3 sequences with reactivity of the two groups of scFvs. Four scFvs, #3, #40, #81 and #83, each representative of one specific CDR3, were selected for further analysis. Competition ELISA experiments showed that they recognize CD152 in its native configuration and bound to different epitopes from the CD80/CD86 interaction site. The scFvs were able to stain human T lymphocytes stimulated either with anti-CD3 and CD28 antibodies or PHA, PMA and ionomycin by cytofluorimetry suggesting that they can be useful reagents for monitoring the kinetics of surface-bound and intracellular CD152.
- Published
- 2000
21. HLA class II alleles associations of anticardiolipin and anti-beta2GPI antibodies in a large series of European patients with systemic lupus erythematosus
- Author
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M, Galeazzi, G D, Sebastiani, A, Tincani, J C, Piette, F, Allegri, G, Morozzi, F, Bellisai, R, Scorza, G B, Ferrara, C, Carcassi, J, Font, G, Passiu, J, Smolen, C, Papasteriades, F, Houssiau, A F, Nebro, E D, Ramon Garrido, A, Jedryka-Goral, and R, Marcolongo
- Subjects
musculoskeletal diseases ,Adult ,Male ,Adolescent ,Enzyme-Linked Immunosorbent Assay ,030204 cardiovascular system & hematology ,Polymerase Chain Reaction ,03 medical and health sciences ,0302 clinical medicine ,Rheumatology ,immune system diseases ,HLA-DQ Antigens ,Humans ,Lupus Erythematosus, Systemic ,skin and connective tissue diseases ,Alleles ,Aged ,Glycoproteins ,030203 arthritis & rheumatology ,Histocompatibility Testing ,DNA ,HLA-DR Antigens ,Middle Aged ,Europe ,Immunoglobulin Isotypes ,beta 2-Glycoprotein I ,Antibodies, Anticardiolipin ,Female ,DNA Probes - Abstract
The objective of this study was to determine the HLA class II associations of the anticardiolipin (aCL) and anti-b2GPI (ab2GPI) antibodies in a large series of European patients with systemic lupus erythematosus (SLE). A cohort of 577 European SLE patients was enrolled. aCL and ab2GPI were measured by ELISA methods. Molecular typing of HLA-DRB1, DRB3, DRB4, DRB5, DQA1 and DQB1 loci was performed by the polymerase chain reaction–sequence specific oligonucleotide probes (PCR–SSOP) method. aCL of IgG, IgM and IgA isotypes were detected in 22.8%, 14% and 13.9% of patients, respectively. IgG and IgM ab2GPI were detected in 20% of patients. aCL showed positive association with HLA DRB1*04, DRB1*0402, DRB1*0403, DRB1*07, DRB3*0301, DQA1*0201, DQA1*0301, DQB1*0302, and negative association with DQA1*0501, DRB3*0202. ab2GPI showed positive association with DRB1*0402, DRB1*0403, DQB1*0302. DRB1*0402 carried the highest relative risk for the presence of both aCL (RR = 8.1) and ab2GPI (RR = 4.6). Our results confirm the already described associations of aCL with HLA DR4 and DR7, but also demonstrate that, among the alleles at the DRB1*04 locus, the *0402 was most represented both in aCL and in ab2GPI positive patients. In addition, HLA class II associations of ab2GPI are for the first time extensively examined in a large cohort of European SLE patients.
- Published
- 2000
22. Anti-ganglioside antibodies in a large cohort of European patients with systemic lupus erythematosus: clinical, serological, and HLA class II gene associations. European Concerted Action on the Immunogenetics of SLE
- Author
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M, Galeazzi, P, Annunziata, G D, Sebastiani, F, Bellisai, V, Campanella, G B, Ferrara, J, Font, F, Houssiau, G, Passiu, E, De Ramon Garrido, A, Fernandez-Nebro, L, Bracci, R, Scorza, P, Puddu, A, Jedryka-Goral, J, Smolen, A, Tincani, C, Carcassi, G, Morozzi, and R, Marcolongo
- Subjects
Adult ,Male ,Adolescent ,Genes, MHC Class II ,Middle Aged ,Cohort Studies ,Europe ,Gangliosides ,Humans ,Lupus Erythematosus, Systemic ,Female ,Nervous System Diseases ,Child ,Aged ,Autoantibodies - Abstract
To assay anti-ganglioside antibodies (aGM1) in sera of a large cohort of European patients with systemic lupus erythematosus (SLE) to define the prevalence of these autoantibodies in SLE; to evaluate the association of aGM1 with clinical manifestations and other autoantibodies found in SLE; and to search for aGM1 association with HLA class II alleles.Four hundred forty-eight patients with SLE were consecutively enrolled in 8 centers from 6 European countries. All sera were tested for antinuclear antibodies by immunofluorescence on HEp-2 cells as substrate, anti-dsDNA, aGM1, aCL, abeta2-glycoprotein I (abeta2-GPI) antibodies by ELISA, and antineutrophil cytoplasmic antibodies (ANCA) by immunofluorescence and by ELISA. Genomic typing for HLA class II loci was performed by polymerase chain reaction-sequence specific oligonucleotide probe method. Clinical assessment was done at the time of enrolment.We found 41.9% of patients with clinical signs of neuropsychiatric involvement; 15.5% of patients were positive for aGM1, 8% of the IgG isotype and 8.6% of the IgM isotype; aGM1-IgG were associated with neuropsychiatric manifestations (NPM) (RR = 3.7), with migraine (RR = 2.4), with OBS (RR = 7.3), and with peripheral neuropathy (RR = 8.5). aGM1-IgM were associated with NPM (RR = 4) and with depression (RR = 3.4). Furthermore, the genetic study showed that aGM1-IgG were associated with HLA-DQB1*0404 (RR = 7.2) while aGM1-IgM were associated with HLA-DQB1*0605 (RR = 33.3). No associations were found between aGM1 and anti-dsDNA, aCL, abeta2GP1, or ANCA.Our results show aGM1 can be found in patients with SLE. aGM1 may play a pathogenetic role for some NPM in this condition.
- Published
- 2000
23. [National quality control of genomic HLA typing]
- Author
-
G B, Ferrara
- Subjects
Quality Control ,Italy ,Histocompatibility Testing ,Histocompatibility Antigens Class I ,Histocompatibility Antigens Class II ,Humans ,Accreditation - Abstract
Since 1996 the Immunogenetics Laboratory of the National Cancer Institute/CBA, Genoa, in collaboration with the Immunohematology Research Centre of Bergamo has organized on behalf of Associazione Italiana di Immunogenetica e Biologia dei Trapianti (AIBT) and Istituto Superiore di Sanità (ISS) the quality control programme of HLA genomic typing for class I and II antigens for the laboratories performing HLA typing for organ and bone marrow transplantations. Herewith are reported the results and comments to the 1997 quality control programme along with some overall data.
- Published
- 2000
24. HLA class I sequence-based typing for high resolution matching in bone marrow transplantation
- Author
-
L, Delfino, A, Longo, A, Morabito, A, Parodi, C, Pera, S, Pozzi, and G B, Ferrara
- Subjects
Histocompatibility Testing ,Humans ,Polymerase Chain Reaction ,Sequence Analysis ,Bone Marrow Transplantation - Published
- 1999
25. HLA-B locus sequence-based typing
- Author
-
S, Pozzi, A, Longo, and G B, Ferrara
- Subjects
Genotype ,HLA-B Antigens ,Chromosome Mapping ,Humans ,Sequence Analysis, DNA ,Polymerase Chain Reaction ,Alleles - Abstract
We describe an approach for HLA-B high-resolution typing. A single locus-specific amplification generates a 1-kb fragment useful for direct sequencing. Four internal primers are necessary for exon 2 and 3 cycle-sequencing in both directions. Fluorescent dye-labelled nucleotides are incorporated during cycle-sequencing and reaction products are analyzed in an automated DNA sequencer. At present, software programs allow automatic assignment of exon 2 only; analysis of exon 3 is not automatic. In the future, the development of more sophisticated software will improve allele assignment. The approach described in this work offers a precise and efficient identification of known allele sequences and at the same time can differentiate new alleles. Furthermore, it may be applied as a model for the development of similar molecular typing approaches for other polymorphic HLA loci.
- Published
- 1999
26. Activation-related differences in HLA class I-bound peptides: presentation of an IL-1 receptor antagonist-derived peptide by activated, but not resting, CD4+ T lymphocytes
- Author
-
G, Frumento, A, Corradi, G B, Ferrara, and A, Rubartelli
- Subjects
CD4-Positive T-Lymphocytes ,Intracellular Fluid ,Antigen Presentation ,Sialoglycoproteins ,Histocompatibility Antigens Class I ,Receptors, Interleukin-1 ,CD8-Positive T-Lymphocytes ,Lymphocyte Activation ,Peptide Fragments ,Interleukin 1 Receptor Antagonist Protein ,Jurkat Cells ,Isomerism ,HLA Antigens ,Humans ,Interphase ,Chromatography, High Pressure Liquid - Abstract
We have compared by reverse phase HPLC the set of peptides eluted from HLA class I molecules in resting and activated CD4+ Jurkat cells. Two peptides were identified that are presented de novo upon activation. After sequencing, one of these peptides turned out to derive from IL-1R antagonist (IL-1Ra). In keeping with this observation, we found that activated, but not resting, Jurkat cells express IL-1Ra. These data indicate that activation of a CD4+ T cell line may result in presentation of peptides derived from proteins expressed de novo after activation. Since IL-1Ra was not known to be expressed by cells of the T lineage, we also investigated its pattern of expression in normal T lymphocytes. Reverse transcriptase-PCR analyses allowed us to demonstrate that IL-1Ra is expressed upon activation by normal CD4+ lymphocytes from peripheral blood and by thymocytes, but not by CD8+ T cells. Interestingly, of the two forms of IL-1Ra that have been detected in different cell lineages, the intracellular one and the secreted one, only the former is expressed by activated CD4+ T cells.
- Published
- 1998
27. Isolation of scFv fragments to polymorphic and monomorphic HLA-DR epitopes from a synthetic phage library
- Author
-
M P, Pistillo, J, Hammer, E, Bono, F, Sinigaglia, A, Gho, C, Marchisone, and G B, Ferrara
- Subjects
Polymorphism, Genetic ,Solubility ,HLA-DR1 Antigen ,Epitopes, B-Lymphocyte ,Humans ,Bacteriophages ,HLA-DR5 Antigen ,Immunoglobulin Fragments ,Gene Library - Abstract
The possibility of producing human recombinant antibodies by the phage display libraries technology has opened up new perspectives in the fields of immunological research and therapeutic applications. Despite the interest for a potential use of this technology in the HLA field, no information is so far available about selection and screening of libraries with HLA antigens. In this study we report our first experience with expression and characterization of human single-chain antibody fragments (scFvs) against HLA-DR epitopes selected from a synthetic phage library.
- Published
- 1997
28. Tipizzazione per sequenza degli antigeni HLA di classe I e II nel trapianto di rene
- Author
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D. Adorno, G. B. Ferrara, A. Canossi, M. Di Rocco, G. Ozzella, I. Contasta, G. Liberatore, F. Papola, I. Monaco, A. Antonacci, L. Delfino, A. Longo, A. Morabito, L. Paleari, C. Pera, and S. Pozzi.
- Subjects
trapianto di rene ,tipizzazione HLA ,SBT - Abstract
Messa a punto della tecnica di sequenziamento degli antigeni HLA, utilizzando i primers locus-specifici per gli esoni d'interesse, al fine di identificare il maggior numero di alleli possibili per una migliore tipizzazione dei pazienti in lista di attesa per trapianto di rene.
- Published
- 1997
29. Reply
- Author
-
Ignazio Olivieri, Angela Padula, Giovanni Ciancio, Carlo Guadiano, Santa Masciandaro, and G. B. Ferrara
- Subjects
Rheumatology ,Immunology ,Immunology and Allergy ,Pharmacology (medical) - Published
- 2003
30. AGE-OF-ONSET IN FAMILIAL ADENOMATOUS POLYPOSIS - HETEROGENEITY WITHIN FAMILIES AND AMONG APC MUTATIONS
- Author
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L. Varesco, Paola Sala, Carlo Rossetti, Silvano Presciuttini, Lucio Bertario, Anna Bafico, G. B. Ferrara, and Viviana Gismondi
- Subjects
Adult ,Male ,congenital, hereditary, and neonatal diseases and abnormalities ,Genes, APC ,Adolescent ,Colorectal cancer ,Molecular Sequence Data ,Pedigree chart ,Biology ,medicine.disease_cause ,Familial adenomatous polyposis ,Genetics ,medicine ,Humans ,Age of Onset ,Child ,Gene ,Genetics (clinical) ,Aged ,Mutation ,Base Sequence ,Genetic Variation ,Middle Aged ,medicine.disease ,Phenotype ,digestive system diseases ,Pedigree ,Adenomatous Polyposis Coli ,Anticipation (genetics) ,Female ,Age of onset ,Colorectal Neoplasms - Abstract
SUMMARY Heterogeneity among and within FAP pedigrees for the age of symptom onset and the age at death from colorectal cancer was studied in a sample of 583 patients of the Italian Polyposis Registry. The among pedigree variation was largely explained by clustering of families in two groups, ‘early FAP’ (most colorectal cancer deaths below 45 years of age) and ‘late FAP’ families (most deaths above age 45). The within-family variation was explained by a marked phenomenon of anticipation (15 years per generation, on the average), possibly not due to ascertainment bias. We then considered the pedigrees with identified mutation in the APC gene. Six families shared a common deletion at codon 1309 and showed the early FAP phenotype. Two families shared a mutation at codon 1061 and revealed the late FAP phenotype. Another two families (codons 453 and 302) clustered with the late FAP group, whereas a family with mutation at codon 835 clustered with the early FAP group. We suggest that there are at least two classes of mutations in the APC gene with different consequences at the phenotypic level. It seems that there are several critical points within the APC protein sequence at which truncation causes a more aggressive disease than truncation at other points.
- Published
- 1994
31. A rapid screening method to detect nonsense and frameshift mutations: identification of disease-causing APC alleles
- Author
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L, Varesco, J, Groden, L, Spirio, M, Robertson, R, Weiss, V, Gismondi, G B, Ferrara, and R, White
- Subjects
Genes, APC ,Base Sequence ,DNA Mutational Analysis ,Molecular Sequence Data ,Humans ,Frameshift Mutation ,Alleles - Abstract
A functional screen for nonsense and frameshift mutations has been devised that allows genes of interest to be scanned in segments. This assay is based on the cloning of these segments in-frame with a colorimetric marker gene (lacZ) followed by screening for the level of functional activity from the marker polypeptide (beta-galactosidase). Individuals at risk for any one of a number of genetic diseases, in particular familial adenomatous polyposis coli (APC), can be quickly screened for chain-terminating mutations introduced by stops and frameshifts. At present, scanning of the APC gene for mutation requires significant effort because it is a large gene and most APC mutations are unique. Therefore, this assay offers a powerful option for the diagnosis of this and other genetic diseases, as well as great potential for the development of a similar rapid screen to detect APC mutations in colorectal adenomas and carcinomas.
- Published
- 1993
32. Molecular oncology: from basic research towards clinical application
- Author
-
G B, Ferrara and L, Santi
- Subjects
Neoplasms ,Research ,Biomarkers, Tumor ,Humans ,DNA, Neoplasm ,Oncogenes ,Molecular Biology - Published
- 1993
33. HLA polymorphism in a Mataco South American Indian tribe: serology of class I and II antigens. Molecular analysis of class II polymorphic variants
- Author
-
C M, Vullo, L, Delfino, G, Angelini, and G B, Ferrara
- Subjects
HLA-D Antigens ,Polymorphism, Genetic ,Haplotypes ,Histocompatibility Testing ,Indians, South American ,Histocompatibility Antigens Class I ,Argentina ,Humans ,Alleles - Abstract
In the present study, HLA-A, B, C, DR, DQ, and DP loci were analyzed in a group of Mataco Amerindians of Argentina. Using reagents from the 11th International Histocompatibility Workshop (11th IHW), class I specifities such as Bw70, Bw75, and Bw48 were found in this population, other than the HLA determinants commonly described in South American Indians. The class II antigens found were DR4, DRw14, and DRw8 at the DR locus, and DQw4 and DQw7 at the DQ locus. The analysis of DRB1-DR4 related alleles, performed by PCR amplification and oligonucleotide probe hybridization, showed the presence of DRB1*0403, *0404, *0405, and *0411 in individuals from this ethnic group. By the analysis of DRB1-DRw14 related alleles, two variants were found: DRB1*1402 and DRB1*1406, the latter provisionally called DRB1 14.6 in 11th IHW. The DRw8-related allele present was DRB1*0802. The analysis of DRB3 gene revealed only the presence of DRB3*0101 allele in DRw14 individuals. DPB1 locus was also analyzed in unrelated individuals of the same population. Only five DPB1 alleles were found: DPB1*0201, *0301, *0402, *0501, and *1301 over the 19 previously described in the literature. These findings emphasize the restricted HLA class I and II variation observed in this ethnic group as it has been previously shown in other American groups. Some particular haplotypes in this Mataco tribe are described in this work.
- Published
- 1992
34. Matching for HLA-DPB1 alleles in zero mismatched HLA-A, -B, and -DR renal transplants
- Author
-
D, Middleton, Y, Mytilineos, D, Savage, G B, Ferrara, G, Angelini, A, Amoroso, F, Trainor, A, Gaweco, G, Mazzola, and L, Delfino
- Subjects
Graft Rejection ,HLA-DP Antigens ,HLA-A Antigens ,Histocompatibility Testing ,HLA-DR Antigens ,Kidney Transplantation ,Survival Rate ,Postoperative Complications ,HLA-B Antigens ,Cadaver ,Humans ,Alleles ,HLA-DP beta-Chains ,Polymorphism, Restriction Fragment Length ,Follow-Up Studies - Published
- 1992
35. Heterogeneous immunophenotype of granular lymphocyte expansions: differential expression of the CD8 alpha and CD8 beta chains
- Author
-
D, de Totero, P L, Tazzari, J P, DiSanto, P F, di Celle, D, Raspadori, R, Conte, M, Gobbi, G B, Ferrara, N, Flomenberg, and F, Lauria
- Subjects
Adult ,Male ,Macromolecular Substances ,CD8 Antigens ,T-Lymphocytes ,CD4-CD8 Ratio ,Receptors, Antigen, T-Cell ,Antibodies, Monoclonal ,Middle Aged ,Lymphoproliferative Disorders ,Immunophenotyping ,Antigens, CD ,Humans ,Female ,Aged - Abstract
In this study, we have evaluated 14 large granular lymphocyte (LGL) expansions, 11 of which were CD8+. Analysis of the membrane expression of the alpha and beta chains of the CD8 antigen, using specific monoclonal antibodies (MoAbs), has shown that LGL expansions with the CD3+, CD4+, CD8+, CD57+ T-cell receptor (TcR) alpha beta phenotype bear the CD8 alpha/alpha isoform, while the CD3+, CD4-, CD8+, CD57+ TcR alpha beta samples were positive for both the CD8 alpha and CD8 beta chains. These data were confirmed also by messenger RNA analysis. One additional case, with a peculiar phenotype (CD3-, CD2-, CD4-, CD8+, CD57-) and a germline configuration of the TcR beta and gamma chain genes, expressed only the CD8 alpha chain. After additional phenotypic analysis with a wider panel of MoAbs, it was found that the beta chain of the interleukin-2 receptor was constitutively expressed on the majority of the samples tested, and that most of the monoclonal samples coexpressed CD45RA/R0 antigens. Using MoAbs directed against the variable regions of the TcR beta chain, we could show a preferential V beta region restriction in the CD8+ monoclonal cases. This more extensive characterization of CD8+ LGL expansions has further documented the marked heterogeneity within this rare condition and allowed a better phenotypic dissection between the monoclonal and polyclonal cases.
- Published
- 1992
36. Analysis of chimerism after bone marrow transplantation using specific oligonucleotide probes
- Author
-
L, Casarino, C, Carbone, M A, Capucci, T, Izzi, and G B, Ferrara
- Subjects
Adult ,Genetic Markers ,Leukemia ,Adolescent ,Base Sequence ,Chimera ,Molecular Sequence Data ,DNA, Neoplasm ,Middle Aged ,Polymerase Chain Reaction ,HLA Antigens ,Humans ,Sibling Relations ,Oligonucleotide Probes ,Polymorphism, Restriction Fragment Length ,Bone Marrow Transplantation - Abstract
DNA hybridization with synthetic oligonucleotide probes was used to follow 18 leukemia patients who received bone marrow transplantation from HLA-identical siblings. Five oligomers complementary to the tandem repetitive sequences of different hypervariable regions of human DNA were designed to produce simple restriction fragment length polymorphism patterns. Each probe hybridized to one or two bands in Hinf I-digested genomic DNA. Combined use of these probes enabled us to distinguish all sibling pairs. DNA analysis early post-transplant (15 days) detected donor-specific fragments in 14 of 18 subjects; two patients had a combination of recipient and donor fragments. Later post-transplant, (102-15 days), one of these two showed only recipient-specific fragments, and the other donor-specific fragments. These data are in accord with other markers of engraftment including cytogenetics and red blood cell phenotyping.
- Published
- 1992
37. European Collection for Biomedical Research
- Author
-
H, Grosse-Wilde and G B, Ferrara
- Subjects
Europe ,HLA Antigens ,Organizations, Nonprofit ,Humans ,Cell Line - Published
- 1992
38. Diverse locations of amino acids in HLA-DR beta chains involved in polymorphic antibody binding epitopes on DR(alpha, beta 1*0101), DR(alpha, beta 1*1101), and DR(alpha,beta 3*0202) molecules
- Author
-
X T, Fu, E, Klohe, C, Alber, W Y, Yu, G B, Ferrara, M P, Pistillo, M, Ballas, and R W, Karr
- Subjects
Epitopes ,Structure-Activity Relationship ,Humans ,HLA-DR Antigens ,Amino Acids - Abstract
In a previous study, we used transfectants expressing hybrid HLA-DR(beta 1*0403)/DR(beta 1*0701) chains to map sequences involved in polymorphic antibody binding epitopes on DR(alpha, beta 1*0403) or DR(alpha, beta 1*0701) molecules. Amino acids 1-40 of the beta 1 domain were found to make the major contributions to most of the antibody binding epitopes studied. To begin to localize sequences that contribute to polymorphic antibody epitopes on DR(alpha,beta 1*0101), DR(alpha,beta 1*1101) and DR(alpha,beta 3*0202) molecules, we used indirect immunofluorescence and flow cytometry to assess the binding of mAb to transfectants expressing hybrid DR(beta 1*0101)/DR(beta 1*1101) or DR(beta 1*1101)/DR(beta 3*0202) chains that divide the DR beta chain into three segments: amino acids 1-40, 41-97, and the beta 2 domain. The results indicate that amino acids 41-97 of the beta 1 domain on DR(beta 1*0101), DR(beta 1*1101), or DR(beta 3*0202) are critical in most of the epitopes, including those recognized by human antibodies MP4 and MP12, and mouse mAb GS88.2, I-LR1, 21r5, and 7.3.19.1, whereas amino acids 1-40 of DR(beta 1*1101) are critical in the epitope recognized by the MCS-7 mAb, and both segments 1-40 and 41-97 of DR(beta 1*1101) are important in the epitopes recognized by the I-LR2 and UL-52 mAbs. Based on these data and comparison of DR beta allelic protein sequences, the residues that may play critical roles in these antibody binding epitopes are predicted.
- Published
- 1992
39. Human allogeneic melanoma-reactive T-helper lymphocyte clones: functional analysis of lymphocyte-melanoma interactions
- Author
-
A. Longo, Meenhard Herlyn, Marina Radrizzani, G. B. Ferrara, Chiara Castelli, Giuseppe Fossati, Giorgio Parmiani, and B. Benedetti
- Subjects
Cytotoxicity, Immunologic ,Cancer Research ,Lymphocyte ,CD3 ,Clone (cell biology) ,Receptors, Antigen, T-Cell ,Biology ,Lymphocyte Activation ,Cell Line ,Antigen ,Antigens, CD ,T-Lymphocyte Subsets ,medicine ,Humans ,IL-2 receptor ,Melanoma ,HLA-D Antigens ,T-cell receptor ,Histocompatibility Antigens Class I ,T lymphocyte ,T-Lymphocytes, Helper-Inducer ,Virology ,Molecular biology ,Clone Cells ,medicine.anatomical_structure ,Oncology ,CD4 Antigens ,biology.protein ,CD8 - Abstract
Lymphocyte clones were isolated from CD4+ peripheral-blood lymphocytes (PBL) of melanoma (Me) patient 9923 (HLA-DR7, DQw2, w6), co-cultured for 30 days with autologous accessory cells, allogeneic Me (Me 1811) (HLA-DR7, DQw1, w2), IL-1 beta (2 U/ml) and IL-2 (15 IU/ml). The 55 clones tested displayed a CD3+, CD4+, CD8-, T-cell receptor (TCR) alpha/beta+, gamma/delta- phenotype. Twenty clones were assayed for proliferation in the presence of Me 1811 and B-lymphoblastoid cell line (LCL) 1811, both expressing HLA-class-I and -II (DR7 and DQw2 shared with patient 9923), intercellular adhesion molecule-1 (ICAM-1) and lymphocyte-function-associated antigen-3 (LFA-3) molecules. Eight clones were found to be reactive to Me 1811 but not to LCL 1811. Specificity analysis of these 8 clones revealed that each of them proliferated only to Me 1811, not to other 14 Me and 12 different LCL, suggesting recognition of melanoma-associated antigen (MAA) expressed on the stimulating Me. One clone (103) was analyzed in more detail. A wider specificity analysis showed that it reacted to Me 1811 but not to 10 other Me expressing or not HLA-DR7, 5 normal melanocyte cultures (2 of them typing HLA-DR7-positive when exposed to interferon-gamma--IFN-gamma), 4 tumors other than Me and 20 different LCL. Clones did not show proliferation in the presence of autologous Me cells. Clone proliferation in response to Me 1811 was significantly inhibited by monoclonal antibodies (MAbs) directed to CD3, TCR alpha/beta, TCR beta chain V12, CD4 and HLA-DR. Moreover, following stimulation with Me 1811, clone 103 showed increased surface expression of CD25 (IL-2 receptor) and CD71 (transferrin receptor) and produced significant amounts of IL-2 and IFN-gamma. The supernatant taken from co-culture of clone 103 with Me 1811 augmented the cytotoxicity of PBL 9923 and other allogeneic PBL against K562 and Me 1811. Thus, the lymphocyte clone 103 is a CD4+ Th clone which uses its CD3/TCR alpha/beta complex to recognize an MAA in conjunction with HLA-DR7. Availability of this type of reagent may prove useful to identify and characterize MAA recognized by T lymphocytes.
- Published
- 1991
40. RFLP analysis with cDNA probes for DQ/DP region in HLA identical sibling marrow transplants: lack of correlation with GvHD
- Author
-
E, Gaozza, P, Piccoli, G B, Ferrara, and A, Bacigalupo
- Subjects
Adult ,Male ,HLA-DP Antigens ,Adolescent ,Genotype ,Graft vs Host Disease ,DNA ,Middle Aged ,Blotting, Southern ,Bone Marrow ,HLA-DQ Antigens ,Humans ,Female ,Oligonucleotide Probes ,Polymorphism, Restriction Fragment Length ,Bone Marrow Transplantation - Abstract
The role of HLA-DP as transplantation antigen in contributing to GvHD is a matter of current debate. HLA-DP, is encoded centromeric to DR-DQ and its alleles are in weak linkage disequilibrium with the rest of the MHC; thus a certain number of HLA matched pairs could be actually DP incompatible to study a possible correlation between HLA-DP matching and GvHD, 24 HLA identical BMT/donor-recipient sibling pairs (serologically tested for HLA Class I and DR antigens) were tested for DQ and DP genes using restriction fragment length polymorphism (RFLP) analysis.DNA extracts were digested according to a standard procedure with two different restriction enzymes (HIND III and MSP I) and hybridised with DQ (alpha and beta) and DP (alpha and beta) specific probes. Highly stringent hybridization and washing conditions were used to prevent cross-hybridizations.Twenty four out of 24 pairs proved to be DQ and DP identical. GvHD developed in 16 out of 24 (66.6%) recipients.These data suggest that DNa analysis of DQ-DP regions, with the probes and enzymes used, does not give predictive informations for GvHD in HLA genotypically identical pairs.
- Published
- 1991
41. Cellular mechanisms of artificial peptides binding to HLA
- Author
-
A Chersi, Guido Frumento, G B Ferrara, Franco Fais, Benvenuto Pernis, and Massimo Vitale
- Subjects
chemistry.chemical_classification ,Viral matrix protein ,Chemistry ,Biomedical Engineering ,Antigen-Presenting Cells ,Medicine (miscellaneous) ,Bioengineering ,Peptide binding ,Peptide ,General Medicine ,Plasma protein binding ,Human leukocyte antigen ,In Vitro Techniques ,Cell biology ,Biomaterials ,Immune system ,HLA Antigens ,Cell culture ,Humans ,Intracellular ,Protein Binding - Abstract
On the basis of the consideration that cell-free models cannot precisely mimic the complexity of the intracellular environment, we used a system to investigate the mechanisms that enable antigen-presenting cells (APC) to bind exogenous peptides through their human leukocyte antigen (HLA) molecules. We evaluated the uptake of the radiolabeled peptide 17-29-Tyr of influenza virus matrix protein by B-EBV cell lines, under various conditions. The results can be summarized as follows: a) the kinetics of peptide binding and release are very fast in living, fully competent cells; b) the peptide-HLA complexes are short-living and the DR molecules continuously undergo peptidic exchange; c) using glutaraldehyde-fixed cells, the kinetics of the two phenomena are slow, closely resembling those observed with the same peptide and purified, immobilized DR molecules. The data suggest that in APC, cellular mechanisms are operative that increase the efficiency of both loading and unloading of Class II HLA with exogenous peptides. This is likely to be related to the recycling of Class II molecules to intracellular compartments, were binding takes place. The observation that the HLA-peptide complex is a dynamic structure, suggests the possibility of replacing natural peptides with synthetic ones at this level, in order to regulate the immune response.
- Published
- 1991
42. Common inactivating APC gene mutations in Italian familial polyposis coli patients
- Author
-
L. Varesco, G. B. Ferrara, Paola Sala, Robert F. James, Viviana Gismondi, Lucio Bertario, M. Ponz de Leon, Paola Grammatico, Guido Biasco, L. De Benedetti, Hugo Aste, and G. Del Porto
- Subjects
Genetics ,Cancer Research ,Oncology ,Epidemiology ,Public Health, Environmental and Occupational Health ,Familial polyposis coli ,Gene mutation ,Biology - Published
- 1993
43. K-ras, p53 and APC mutations in sporadic colorectal adenomas
- Author
-
Viviana Gismondi, Guglielmina Nadia Ranzani, N. S. Pellegata, L. De Benedetti, L. Varesco, Hugo Aste, Luigina Bonelli, G. B. Ferrara, R. James, Stefania Sciallero, and Lorena Losi
- Subjects
Cancer Research ,Oncology ,Epidemiology ,Public Health, Environmental and Occupational Health - Published
- 1993
44. PCR-SSCP analysis of the APC c-ene in Italian familial polyposis coli patients
- Author
-
Hugo Aste, G. Biasco, Paola Grammatico, G. DelPorto, R. Sassatelli, A. Allegretti, G. B. Ferrara, Liliana Varesco, Pere Sala, Lucio Bertario, M. Ponzdeleon, R. James, M.T. Illeni, L. DeBenedetti, Viviana Gismondi, and L. Casarino
- Subjects
Cancer Research ,Pcr sscp ,Genetics ,Familial polyposis coli ,Biology ,Molecular Biology ,Molecular biology ,Ene reaction - Published
- 1992
45. Cytogenetic and molecular study of 30 malignant melanoma primary cell cultures
- Author
-
Ugo Bottoni, L. Casarino, Viviana Gismondi, Paola Grammatico, Barbara Grammatico, L. De Benedetti, F. Roccella, L. Varesco, G. B. Ferrara, Abdelhamid Heouaine, M. Roccella, G. Del Porto, and O.A. Carlesimo
- Subjects
Primary (chemistry) ,Oncology ,business.industry ,Cell culture ,Melanoma ,Cancer research ,Medicine ,business ,medicine.disease - Published
- 1991
46. Analysis of HLA-DP allelic sequence polymorphism using the in vitro enzymatic DNA amplification of DP-alpha and DP-beta loci
- Author
-
T L Bugawan, G T Horn, C M Long, E Mickelson, J A Hansen, G B Ferrara, G Angelini, and H A Erlich
- Subjects
Immunology ,Immunology and Allergy - Abstract
Allelic sequence variation of the HLA DP-alpha and DP-beta genes has been analyzed in a panel of 34 DP-typed cell lines. The polymorphic second exon of these genes was specifically amplified in vitro by the polymerase chain reaction method, using the thermostable DNA polymerase of Thermus, aquaticus. The analysis of M13 clones containing the amplified DP-beta sequences revealed a total of 14 allelic variants. In general, specific allelic DP-beta sequences were associated with each of the defined DPw1-w6 types, with beta allele subtypes revealed for the DPw2 and DPw4 specificities. An additional six DP-beta alleles which did not correlate with any of the T cell-defined specificities (DP "blanks") were also identified. Only the two previously characterized alleles of DP-alpha were detected. These observations suggest that the T cell-defined DP specificities are determined by polymorphic residues on the beta-chain. The sequence polymorphisms in DP-beta are clustered in a few specific regions, and can be detected using sequence-specific oligonucleotide probes and polymerase chain reaction amplified DNA in a rapid dot-blot format. This approach provides a simple and informative method of DP typing. The DP-beta sequences derived from four DP-typed celiac disease patients were compared with the distribution of DP-beta alleles in control individuals.
- Published
- 1988
47. WORKSHOP A: STRUCTURE AND FUNCTION OF MHC CLASS I AND CLASS IV MOLECULES
- Author
-
G. B. Ferrara and Jonathan C. Howard
- Subjects
Class (set theory) ,biology ,Immunology ,MHC class I ,Genetics ,biology.protein ,Computational biology ,Structure and function - Published
- 1987
48. Serologically detectable polymorphism of the HLA-DC alpha-subunit
- Author
-
R Tosi, N Tanigaki, R Sorrentino, D Centis, and G B Ferrara
- Subjects
Immunology ,Immunology and Allergy - Abstract
Two 8th Workshop alloantisera, 8W1062 and 8W614, which were classified respectively as anti-DR5 and anti-DR3, were found to react with an Ia preparation from LG38 cells (DR5,5) that was depleted of DR and BR molecules and enriched in DC molecules by pretreatment with a monoclonal antibody and an antiserum against alpha-subunits of DR and BR molecules. The reaction of both alloantisera was inhibited by DR3-positive, DR5-positive, and DRW13-positive individuals. This pattern does not correspond to any of the hitherto reported supertypic specificities. Both antisera were shown to bind to the alpha-subunit isolated from the DC-enriched preparation but not to the beta-subunit. The specificity has been named DC alpha 3. Family studies show the segregation according to the HLA haplotype patterns. This demonstrates by formal genetics criteria the HLA linkage of the locus controlling the DC alpha-subunit.
- Published
- 1984
49. The Merrit Alloantigenic System of Human B Lymphocytes: Evidence for Thirteen Possible Factors including one Six-Member Segregant Series
- Author
-
Roy L. Walford, Thomas Gossett, G. M. Troup, Richard A. Gatti, Kay K. Mittal, A. Robins, G. B. Ferrara, and Edith Zeller
- Subjects
Immunology ,Immunology and Allergy - Abstract
An analysis of the typing results of a 70-member chronic lymphatic leukemia B cell panel revealed evidence for 13 possible groups of the Merrit alloantigenic system. Six of these appeared possibly allelic and may represent a segregant series. The CLL panel was also fully typed for HLA and some degree of linkage dysequilibrium between Merrit and HLA seemed apparent from the data. Merrit antibodies can be absorbed out with selected surface membrane immunoglobulin (SMIG)-positive normal lymphocytes and less so or not at all with E rosette-forming T cells or Fc-positive SMIG-negative lymphocytes
- Published
- 1976
50. Radioimmunoassay typing gives a more precise definition of the HLA association of Type I (insulin-dependent) diabetes
- Author
-
D. Maccarone, A. Candela, M. Vela, M. P. Raponi, D. Adorno, Roberto Tosi, M. Orsini, Nobuyuki Tanigaki, L. Campea, D. Centis, A. Longo, G. B. Ferrara, and F. Papola
- Subjects
musculoskeletal diseases ,Genetics ,Linkage disequilibrium ,Endocrinology, Diabetes and Metabolism ,Radioimmunoassay ,Human physiology ,Hla association ,Biology ,medicine.disease ,Antigen ,immune system diseases ,Insulin dependent diabetes ,Diabetes mellitus ,Immunology ,Internal Medicine ,medicine ,Typing ,skin and connective tissue diseases - Abstract
The problem of the HLA association of Type 1 (insulin-dependent) diabetes was re-examined by testing Class II antigenic specificities detectable by radioimmunoassay. Established (DRw53, DQwl, DQw2, DQw3) as well as newly described (DC5, DCalpha3) specificities were typed. The data obtained suggest that the association with DR3 and DR4 is secondary to that with DQ specificities in linkage disequilibrium with DR3 and DR4.
- Published
- 1986
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