20 results on '"Imperiali, Fg"'
Search Results
2. Sustained response to low-dose rituximab in idiopathic autoimmune hemolytic anemia
- Author
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Barcellini, W, Zaja, Francesco, Zaninoni, A, Imperiali, Fg, Di Bona, E, Fattizzo, B, Consonni, D, Cortelezzi, A, Zanella, A., Barcellini, W, Zaja, Francesco, Zaninoni, A, Imperiali, Fg, Di Bona, E, Fattizzo, B, Consonni, D, Cortelezzi, A, and Zanella, A.
- Abstract
ObjectivesTo evaluate the sustained response to low-dose (LD) rituximab in autoimmune hemolytic anemia (AIHA), the ex vivo effect on anti-RBC antibody production by mitogen-stimulated direct antiglobulin test (MS-DAT), and the in vitro dose effect of the drug on the production of anti-RBC antibodies. MethodsThirty two patients, 18 warm (W) AIHA and 14 cold hemagglutinin disease (CHD), were treated with LD rituximab (100mg fixed dose x4 weekly infusions) along with a short course of oral prednisone. Complete clinical examination, blood counts, and hemolytic markers were performed at enrollment and at month 6, 12, 24, and 36. ResultsHematological parameters significantly improved at all time points compared to enrollment. The overall response was 90%, 100%, 100%, and 89% and the relapse-free survival 87%, 79%, 68%, and 68% at 6, 12, 24, and 36months, respectively. Response rates were slightly better in WAIHA than in CHD, and relapse risk was greater in cold than warm forms (HR 2.1, 95% CI 0.6-7.9). Four patients were retreated (one patient twice), all achieving a response, lasting a median of 18months (range 9-30). Treatment was well tolerated without adverse events or infections. Anti-RBC antibody production by MS-DAT significantly decreased over time. In vitro studies showed that rituximab effectively inhibited anti-RBC antibody production at 50g/mL, 1/6 of the drug concentration after therapy with standard doses. ConclusionsThese data confirm that LD rituximab treatment is effective and induces sustained responses in AIHA, and that a lower dose of the drug is enough to down-regulate autoantibody production.
- Published
- 2013
3. Low-dose rituximab in adult patients with idiopathic autoimmune hemolytic anemia: clinical efficacy and biologic studies
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Eros Di Bona, Dario Consonni, Marta Lisa Battista, Renato Fanin, Wilma Barcellini, Anna Zaninoni, Francesca Guia Imperiali, Bruno Fattizzo, Alberto Zanella, Agostino Cortelezzi, Francesco Zaja, Barcellini, W, Zaja, Francesco, Zaninoni, A, Imperiali, Fg, Battista, Ml, Di Bona, E, Fattizzo, B, Consonni, D, Cortelezzi, A, Fanin, Renato, and Zanella, A.
- Subjects
Hemolytic anemia ,Adult ,Male ,medicine.medical_specialty ,Anemia ,Immunology ,Biochemistry ,Gastroenterology ,Disease-Free Survival ,Antibodies, Monoclonal, Murine-Derived ,Pharmacotherapy ,Risk Factors ,Internal medicine ,medicine ,Secondary Prevention ,Humans ,Immunologic Factors ,Prospective Studies ,Adverse effect ,Aged ,Biologic marker ,Dose-Response Relationship, Drug ,Cumulative dose ,business.industry ,Cell Biology ,Hematology ,Middle Aged ,medicine.disease ,Treatment Outcome ,Cytokines ,Rituximab ,Drug Therapy, Combination ,Female ,Steroids ,Anemia, Hemolytic, Autoimmune ,Autoimmune hemolytic anemia ,business ,medicine.drug - Abstract
This prospective study investigated the efficacy, safety, and response duration of low-dose rituximab (100 mg fixed dose for 4 weekly infusions) together with a short course of steroids as first- or second-line therapy in 23 patients with primary autoimmune hemolytic anemia (AIHA). The overall response was 82.6% at month +2, and subsequently stabilized to ∼ 90% at months +6 and +12; the response was better in warm autoimmune hemolytic anemia (WAIHA; overall response, 100% at all time points) than in cold hemagglutinin disease (CHD; average, 60%); the relapse-free survival was 100% for WAIHA at +6 and +12 months versus 89% and 59% in CHD, respectively, and the estimated relapse-free survival at 2 years was 81% and 40% for the warm and cold forms, respectively. The risk of relapse was higher in CHD and in patients with a longer interval between diagnosis and enrollment. Steroid administration was reduced both as cumulative dose (∼ 50%) and duration compared with the patient's past history. Treatment was well tolerated and no adverse events or infections were recorded; retreatment was also effective. The clinical response was correlated with amelioration biologic markers such as cytokine production (IFN-γ, IL-12, TNF-α, and IL-17), suggesting that low-dose rituximab exerts an immunomodulating activity. This study is registered at www.clinicaltrials.gov as NCT01345708.
- Published
- 2012
4. Characterization of Transport Activity of SLC11 Transporters in Xenopus laevis Oocytes by Fluorophore Quenching.
- Author
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Cinquetti R, Imperiali FG, Bozzaro S, Zanella D, Vacca F, Roseti C, Peracino B, Castagna M, and Bossi E
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- Animals, Biological Transport, Cation Transport Proteins metabolism, Cation Transport Proteins physiology, Dictyostelium metabolism, Female, Fluoresceins pharmacokinetics, Fluorescent Dyes pharmacokinetics, Membrane Potentials, Microscopy, Fluorescence, Oocytes chemistry, Oocytes metabolism, Xenopus laevis, Electrophysiological Phenomena, Membrane Transport Proteins metabolism, Patch-Clamp Techniques methods
- Abstract
Membrane proteins are involved in different physiological functions and are the target of pharmaceutical and abuse drugs. Xenopus laevis oocytes provide a powerful heterologous expression system for functional studies of these proteins. Typical experiments investigate transport using electrophysiology and radiolabeled uptake. A two-electrode voltage clamp is suitable only for electrogenic proteins, and uptake measurements require the existence of radiolabeled substrates and adequate laboratory facilities.Recently, Dictyostelium discoideum Nramp1 and NrampB were characterized using multidisciplinary approaches. NrampB showed no measurable electrogenic activity, and it was investigated in Xenopus oocytes by acquiring confocal images of the quenching of injected fluorophore calcein.This method is adequate to measure the variation in emitted fluorescence, and thus transporter activity indirectly, but requires long experimental procedures to collect statistically consistent data. Considering that optimal expression of heterologous proteins lasts for 48-72 h, a slow acquiring process requires the use of more than one batch of oocytes to complete the experiments. Here, a novel approach to measure substrate uptake is reported. Upon injection of a fluorophore, oocytes were incubated with the substrate and the transport activity measured, evaluating fluorescence quenching in a microplate reader. The technique permits the testing of tens of oocytes in different experimental conditions simultaneously, and thus the collection of significant statistical data for each batch, saving time and animals.The method was tested with different metal transporters (SLC11), DMT1, Dd Nramp1, and Dd NrampB, and verified with the peptide transporter PepT1 (SLC15). Comparison with traditional methods (uptake, two-electrode voltage clamp) and with quenching images acquired by fluorescence microscopy confirmed its efficacy.
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- 2021
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5. Detection of erythroblast antibodies in mitogen-stimulated bone marrow cultures from patients with myelodysplastic syndromes.
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Zaninoni A, Imperiali FG, Cattaneo A, Soverini G, Binda F, Porretti L, Cortelezzi A, and Barcellini W
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- Adult, Aged, Aged, 80 and over, Apoptosis drug effects, Erythropoietin immunology, Female, Humans, Immunoglobulin G immunology, Male, Middle Aged, Antibodies immunology, Bone Marrow Cells drug effects, Bone Marrow Cells immunology, Erythroblasts drug effects, Erythroblasts immunology, Mitogens pharmacology, Myelodysplastic Syndromes immunology
- Abstract
Background: Low-risk myelodysplastic syndromes (MDS) show several immunologic abnormalities, including increased frequency of autoimmune manifestations and/or overt autoimmune diseases, whose prognostic significance still remains controversial., Study Design and Methods: We studied the presence of erythroblast antibodies in mitogen-stimulated bone marrow (BM) cultures of 70 patients with early-stage MDS (refractory anemia and refractory anemia with ringed sideroblasts)., Results: Sixty-six percent of patients showed positive erythroblast antibodies, along with BM erythroid hyperplasia and a hemolytic picture in the peripheral blood. Supernatants from positive cultures induced an increase of overall cellularity, the appearance of erythroblastic clustering, and dyserythropoietic signs in normal BM. We identified CD45(dim) Gly-A(dim) CD71(bright) cells (red blood cell precursors at different maturation stage) as the target of the antibodies. Erythropoietin (EPO) levels were reduced and EPO receptors (EPO-R) increased in BM culture supernatants from positive patients. However, flow cytometric analysis showed that neither EPO nor EPO-R was involved in an abnormal stimulation driven by these autoantibodies. Values of the proapoptotic protein Bax were increased in positive patients and Bcl-2 levels were decreased, although not significantly., Conclusion: MDS patients with anti-erythroblast autoimmunity showed increased BM apoptosis, suggesting that the autoimmune reaction may contribute to an unfavorable BM microenvironment for optimal erythropoiesis., (© 2016 AABB.)
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- 2016
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6. Amino acid transporter B(0)AT1 (slc6a19) and ancillary protein: impact on function.
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Margheritis E, Imperiali FG, Cinquetti R, Vollero A, Terova G, Rimoldi S, Girardello R, and Bossi E
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- Animals, Bass metabolism, Biological Transport physiology, Carrier Proteins metabolism, Humans, Intestinal Mucosa metabolism, Oocytes metabolism, Salmo salar metabolism, Substrate Specificity, Xenopus laevis metabolism, Amino Acid Transport Systems metabolism, Amino Acid Transport Systems, Neutral metabolism, Amino Acids metabolism
- Abstract
Amino acids play an important role in the metabolism of all organisms. Their epithelial re-absorption is due to specific transport proteins, such as B(0)AT1, a Na(+)-coupled neutral amino acid symporter belonging to the solute carrier 6 family. Here, a recently cloned fish orthologue, from the intestine of Salmo salar, was electrophysiologically characterized with the two-electrode voltage clamp technique, in Xenopus laevis oocytes heterologously expressing the transporter. Substrate specificity, apparent affinities and the ionic dependence of the transport mechanism were determined in the presence of specific collectrin. Results demonstrated that like the human, but differently from sea bass (Dicentrarchus labrax) orthologue, salmon B(0)AT1 needs to be associated with partner proteins to be correctly expressed at the oocyte plasma membrane. Cloning of sea bass collectrin and comparison of membrane expression and functionality of the B(0)AT1 orthologue transporters allowed a deeper investigation on the role of their interactions. The parameters acquired by electrophysiological and immunolocalization experiments in the mammalian and fish transporters contributed to highlight the dynamic of relations and impacts on transport function of the ancillary proteins. The comparative characterization of the physiological parameters of amino acid transporters with auxiliary proteins can help the comprehension of the regulatory mechanism of essential nutrient absorption.
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- 2016
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7. The D-amino acid transport by the invertebrate SLC6 transporters KAAT1 and CAATCH1 from Manduca sexta.
- Author
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Vollero A, Imperiali FG, Cinquetti R, Margheritis E, Peres A, and Bossi E
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- Animals, Isomerism, Patch-Clamp Techniques, Amino Acid Transport Systems, Neutral metabolism, Amino Acids metabolism, Carrier Proteins metabolism, Insect Proteins metabolism, Manduca metabolism, Membrane Proteins metabolism, Protein Transport physiology
- Abstract
The ability of the SLC6 family members, the insect neutral amino acid cotransporter KAAT1(K(+)-coupled amino acid transporter 1) and its homologous CAATCH1(cation anion activated amino acid transporter/channel), to transport D-amino acids has been investigated through heterologous expression in Xenopus laevis oocytes and electrophysiological techniques. In the presence of D-isomers of leucine, serine, and proline, the msKAAT1 generates inward, transport-associated, currents with variable relative potencies, depending on the driving ion Na(+) or K(+). Higher concentrations of D-leucine (≥1 mmol/L) give rise to an anomalous response that suggests the existence of a second binding site with inhibitory action on the transport process. msCAATCH1 is also able to transport the D-amino acids tested, including D-leucine, whereas L-leucine acts as a blocker. A similar behavior is exhibited by the KAAT1 mutant S308T, confirming the relevance of the residue in this position in L-leucine binding and the different interaction of D-leucine with residues involved in transport mechanism. D-leucine and D-serine on various vertebrate orthologs B(0)AT1 (SLC6A19) elicited only a very small current and singular behavior was not observed, indicating that it is specific of the insect neutral amino acid transporters. These findings highlight the relevance of D-amino acid absorption in the insect nutrition and metabolism and may provide new evidences in the molecular transport mechanism of SLC6 family., (© 2016 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.)
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- 2016
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8. Detection of red blood cell antibodies in mitogen-stimulated cultures from patients with hereditary spherocytosis.
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Zaninoni A, Vercellati C, Imperiali FG, Marcello AP, Fattizzo B, Fermo E, Bianchi P, Grossi C, Cattaneo A, Cortelezzi A, Zanella A, and Barcellini W
- Subjects
- Adolescent, Adult, Aged, Aged, 80 and over, Cellular Senescence, Child, Child, Preschool, Coombs Test, Female, Humans, Immunoglobulin G blood, Male, Middle Aged, Mitogens pharmacology, Autoantibodies blood, Erythrocytes immunology, Spherocytosis, Hereditary immunology
- Abstract
Background: Hereditary spherocytosis (HS) is a congenital hemolytic anemia caused by defects in red blood cell (RBC) membrane proteins leading to premature RBC clearance in the spleen. The presence of RBC autoantibodies has never been extensively investigated in HS., Study Design and Methods: RBC antibody-bound immunoglobulin (Ig)G was investigated in 91 consecutive HS patients by mitogen-stimulated direct antiglobulin test (MS-DAT), a sensitive method able to magnify latent RBC antibody autoimmunity and related with hemolytic variables, previous splenectomy, and type of membrane defect., Results: A total of 61% of HS cases had RBC antibodies by MS-DAT (29 Band 3, 17 spectrin deficiency, and nine no defined defect). The amount of RBC-bound IgG was greater in HS compared with controls (236 ± 192 ng/mL vs. 52 ± 29 ng/mL, p < 0.0001), although lower than that observed in autoimmune hemolytic anemia (AIHA; 634 ± 371 ng/mL vs. 236 ± 192 ng/mL, p < 0.0001). Western blot experiments showed that purified IgG fraction from MS-DAT-positive patients bind to α- and β-spectrin, Band 3, and Band 4.9. Positive cases displayed increased reticulocytosis and slightly reduced hemoglobin (Hb) values compared to negative ones. Patients displaying RBC-bound IgG of more than 250 ng/mL (the positive threshold of AIHA) showed increased number of spherocytes and mainly had spectrin deficiency. RBC-bound IgG and free Hb increased over time after storage at 4°C, a surrogate of ex vivo aging, more evidently in HS than controls, and particularly in Band 3 deficiency., Conclusion: RBC autoantibodies were detected by MS-DAT in more than a half of HS patients. Positive cases showed a more evident hemolytic pattern suggesting a pathogenic role of these autoantibodies in RBC opsonization and splenic removal., (© 2015 AABB.)
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- 2015
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9. Toll-like receptor 4 and 9 expression in B-chronic lymphocytic leukemia: relationship with infections, autoimmunity and disease progression.
- Author
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Barcellini W, Imperiali FG, Zaninoni A, Reda G, Consonni D, Fattizzo B, Lonati S, Nobili L, Zanella A, and Cortelezzi A
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- Aged, Aged, 80 and over, Autoimmunity, Biomarkers metabolism, Disease Progression, Female, Humans, Infections etiology, Leukemia, Lymphocytic, Chronic, B-Cell complications, Leukemia, Lymphocytic, Chronic, B-Cell diagnosis, Leukemia, Lymphocytic, Chronic, B-Cell immunology, Leukemia, Lymphocytic, Chronic, B-Cell mortality, Leukemia, Lymphocytic, Chronic, B-Cell therapy, Male, Middle Aged, Neoplasm Staging, Prognosis, Retrospective Studies, Toll-Like Receptor 4 metabolism, Toll-Like Receptor 9 metabolism, Treatment Outcome, Gene Expression, Leukemia, Lymphocytic, Chronic, B-Cell genetics, Toll-Like Receptor 4 genetics, Toll-Like Receptor 9 genetics
- Abstract
Toll-like receptors (TLRs) represent major agents of innate immunity and initiators of adaptive immunity. TLR4 and TLR9 gene expression was related to the occurrence of infections, autoimmunity and disease progression in 95 patients with B-chronic lymphocytic leukemia (B-CLL), grouped according to stage, therapy and known prognostic markers, and followed prospectively (median 33.6 months, range 25-50). A retrospective analysis (median 6.8 years, range 6-26) was also performed. TLR4 gene expression was decreased and TLR9 increased in patients versus controls, the former being more pronounced in advanced and multi-treated disease, and in patients with unmutated immunoglobulin heavy chain variable (IgVH) region and unfavorable cytogenetics. Patients with reduced TLR4 had an increased risk of disease progression and development of autoimmune complications. No relationship was found between reduced TLR4 expression and infectious episodes, which were observed in advanced stages and treated patients. These findings suggest that impaired innate immunity identifies patients with B-CLL with a poor prognosis and reduced ability to silence autoreactive phenomena.
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- 2014
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10. Sustained response to low-dose rituximab in idiopathic autoimmune hemolytic anemia.
- Author
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Barcellini W, Zaja F, Zaninoni A, Imperiali FG, Di Bona E, Fattizzo B, Consonni D, Cortelezzi A, and Zanella A
- Subjects
- Adult, Aged, Anemia, Hemolytic, Autoimmune diagnosis, Anemia, Hemolytic, Autoimmune mortality, Female, Humans, Male, Middle Aged, Recurrence, Rituximab, Time Factors, Treatment Outcome, Young Adult, Anemia, Hemolytic, Autoimmune drug therapy, Antibodies, Monoclonal, Murine-Derived administration & dosage, Immunologic Factors administration & dosage
- Abstract
Objectives: To evaluate the sustained response to low-dose (LD) rituximab in autoimmune hemolytic anemia (AIHA), the ex vivo effect on anti-RBC antibody production by mitogen-stimulated direct antiglobulin test (MS-DAT), and the in vitro dose effect of the drug on the production of anti-RBC antibodies., Methods: Thirty two patients, 18 warm (W) AIHA and 14 cold hemagglutinin disease (CHD), were treated with LD rituximab (100 mg fixed dose ×4 weekly infusions) along with a short course of oral prednisone. Complete clinical examination, blood counts, and hemolytic markers were performed at enrollment and at month 6, 12, 24, and 36., Results: Hematological parameters significantly improved at all time points compared to enrollment. The overall response was 90%, 100%, 100%, and 89% and the relapse-free survival 87%, 79%, 68%, and 68% at 6, 12, 24, and 36 months, respectively. Response rates were slightly better in WAIHA than in CHD, and relapse risk was greater in cold than warm forms (HR 2.1, 95% CI 0.6-7.9). Four patients were retreated (one patient twice), all achieving a response, lasting a median of 18 months (range 9-30). Treatment was well tolerated without adverse events or infections. Anti-RBC antibody production by MS-DAT significantly decreased over time. In vitro studies showed that rituximab effectively inhibited anti-RBC antibody production at 50 μg/mL, 1/6 of the drug concentration after therapy with standard doses., Conclusions: These data confirm that LD rituximab treatment is effective and induces sustained responses in AIHA, and that a lower dose of the drug is enough to down-regulate autoantibody production., (© 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)
- Published
- 2013
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11. Increased prevalence of autoimmune phenomena in myelofibrosis: relationship with clinical and morphological characteristics, and with immunoregulatory cytokine patterns.
- Author
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Barcellini W, Iurlo A, Radice T, Imperiali FG, Zaninoni A, Fattizzo B, Guidotti F, Bianchi P, Fermo E, Consonni D, and Cortelezzi A
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- Adult, Aged, Aged, 80 and over, Coombs Test, Female, Follow-Up Studies, Humans, Interleukin-17 metabolism, Interleukin-8 metabolism, Male, Middle Aged, Prevalence, Primary Myelofibrosis epidemiology, Primary Myelofibrosis metabolism, Prognosis, Transforming Growth Factor beta metabolism, Antibodies, Anti-Idiotypic immunology, Autoimmunity immunology, Cytokines metabolism, Erythrocytes immunology, Primary Myelofibrosis immunology
- Abstract
Autoimmune phenomena and cytokines were investigated in 100 patients with myelofibrosis (MF) and related to marrow fibrosis and clinical risk. Anti-erythrocyte antibodies by mitogen-stimulated direct antiglobulin test (MS-DAT) were positive in 45%, anti-platelets in 15% and organ/non organ-specific in 57% of cases, without clinically overt disease, and mostly in low-risk/intermediate-risk-1 and MF-0/MF-1. TGF-β and IL-8 were increased in MS-DAT positive cases, and IFN-γ in patients with serological autoantibodies. TGF-β and IL-17 were elevated in early clinical and morphological stages, while IL-8 increased in advanced stages. These data suggest that autoimmune phenomena and cytokine disregulation are particularly relevant in early MF., (Copyright © 2013 Elsevier Ltd. All rights reserved.)
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- 2013
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12. Low-dose rituximab in adult patients with idiopathic autoimmune hemolytic anemia: clinical efficacy and biologic studies.
- Author
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Barcellini W, Zaja F, Zaninoni A, Imperiali FG, Battista ML, Di Bona E, Fattizzo B, Consonni D, Cortelezzi A, Fanin R, and Zanella A
- Subjects
- Adult, Aged, Anemia, Hemolytic, Autoimmune epidemiology, Anemia, Hemolytic, Autoimmune immunology, Antibodies, Monoclonal, Murine-Derived adverse effects, Cytokines blood, Cytokines immunology, Disease-Free Survival, Dose-Response Relationship, Drug, Drug Therapy, Combination, Female, Humans, Immunologic Factors adverse effects, Male, Middle Aged, Prospective Studies, Risk Factors, Rituximab, Secondary Prevention, Steroids administration & dosage, Steroids adverse effects, Treatment Outcome, Anemia, Hemolytic, Autoimmune drug therapy, Antibodies, Monoclonal, Murine-Derived administration & dosage, Immunologic Factors administration & dosage
- Abstract
This prospective study investigated the efficacy, safety, and response duration of low-dose rituximab (100 mg fixed dose for 4 weekly infusions) together with a short course of steroids as first- or second-line therapy in 23 patients with primary autoimmune hemolytic anemia (AIHA). The overall response was 82.6% at month +2, and subsequently stabilized to ∼ 90% at months +6 and +12; the response was better in warm autoimmune hemolytic anemia (WAIHA; overall response, 100% at all time points) than in cold hemagglutinin disease (CHD; average, 60%); the relapse-free survival was 100% for WAIHA at +6 and +12 months versus 89% and 59% in CHD, respectively, and the estimated relapse-free survival at 2 years was 81% and 40% for the warm and cold forms, respectively. The risk of relapse was higher in CHD and in patients with a longer interval between diagnosis and enrollment. Steroid administration was reduced both as cumulative dose (∼ 50%) and duration compared with the patient's past history. Treatment was well tolerated and no adverse events or infections were recorded; retreatment was also effective. The clinical response was correlated with amelioration biologic markers such as cytokine production (IFN-γ, IL-12, TNF-α, and IL-17), suggesting that low-dose rituximab exerts an immunomodulating activity. This study is registered at www.clinicaltrials.gov as NCT01345708.
- Published
- 2012
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13. Hereditary red cell membrane defects: diagnostic and clinical aspects.
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Barcellini W, Bianchi P, Fermo E, Imperiali FG, Marcello AP, Vercellati C, Zaninoni A, and Zanella A
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- Humans, Spherocytosis, Hereditary diagnosis, Spherocytosis, Hereditary genetics, Spherocytosis, Hereditary pathology, Spherocytosis, Hereditary therapy, Elliptocytosis, Hereditary diagnosis, Elliptocytosis, Hereditary genetics, Elliptocytosis, Hereditary pathology, Elliptocytosis, Hereditary therapy, Erythrocyte Membrane genetics, Erythrocyte Membrane pathology, Erythrocytes, Abnormal pathology
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- 2011
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14. Comparison of traditional methods and mitogen-stimulated direct antiglobulin test for detection of anti-red blood cell autoimmunity.
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Barcellini W, Revelli N, Imperiali FG, Villa MA, Manera MC, Paccapelo C, Zaninoni A, and Zanella A
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- Adult, Aged, Aged, 80 and over, Female, Humans, Male, Middle Aged, Young Adult, Anemia, Hemolytic, Autoimmune diagnosis, Autoantibodies immunology, Autoimmunity, Coombs Test methods, Erythrocytes immunology
- Abstract
The diagnosis of autoimmune hemolytic anemia (AIHA) is based on a positive direct antiglobulin test (DAT), which is performed using various methods with different sensitivities. Recently, mitogen-stimulated (MS)-DAT was suggested to be able to identify latent anti-erythrocyte autoimmunity. Traditional methods (tube, microcolumn, and solid phase) and MS-DAT were compared in 54 consecutive cases of suspected AIHA, 28 idiopathic AIHA in clinical remission, and 12 difficult-to-diagnose cases of DAT-negative AIHA, and the results (all cases) were correlated with hematologic and hemolytic parameters. DAT tube was confirmed as the gold standard to diagnose AIHA since almost all positive cases showed hemolytic anemia and positive eluates; 10 out of 26 tube-negative cases were positive on microcolumn and solid phase antiglobulin tests, and 22 out of 26 using MS-DAT, although only half of them showed clear signs of hemolysis. Mitogen stimulation increased the amount of IgG bound to red blood cells in all groups; moreover, MS-DAT was the only positive test in 10 cases of AIHA, and mitogen stimulation facilitated the identification of autoantibody specificity in culture supernatants. We conclude that a battery of tests rather than a single test is useful for the diagnosis of AIHA, including MS-DAT as an additional test for selected cases, although the results have to be cautiously interpreted based on the overall clinical context.
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- 2010
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15. Bone marrow mitogen-stimulated direct antiglobulin test in a case of erythroblastic synartesis.
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Zaninoni A, Imperiali FG, Pomati M, Colombi M, Boschetti C, and Barcellini W
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- Adult, Anemia immunology, Anemia pathology, Anemia, Hemolytic pathology, Antibodies, Anti-Idiotypic immunology, Bone Marrow Cells immunology, Bone Marrow Cells pathology, Erythroblasts immunology, Erythroblasts pathology, Humans, Killer Cells, Natural immunology, Killer Cells, Natural pathology, Male, T-Lymphocytes immunology, T-Lymphocytes pathology, Anemia, Hemolytic immunology, Cytotoxicity Tests, Immunologic methods, Erythropoiesis immunology
- Abstract
In this article we report a case of erythroblastic synartesis, a rare disease characterized by ineffective erythropoiesis, clusters of erythroblasts due to membrane invaginations, in which an autoimmune pathogenesis is hypothesized. We investigated the presence of anti-erythroblast autoimmunity in bone marrow cultures using a mitogen-stimulated direct antiglobulin test, a method reported to be able to disclose a latent autoimmunity in various diseases. The test revealed the presence of erythroblast-bound IgG, supporting the hypothesis of the autoimmune pathogenesis of erythroblastic synartesis. Supernatants induced the same specific morphological features, i.e erythroblastic clustering and diserythropoietic signs (multiple nuclei, nuclear inclusions, and intercellular bridges) in normal progenitors.
- Published
- 2010
16. Hematological, molecular and cytokine changes after reduced intensity bone marrow transplantation for paroxysmal nocturnal hemoglobinuria.
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Barcellini W, Fermo E, Imperiali FG, Zaninoni A, Boschetti C, Onida F, and Soligo D
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- Adult, Case-Control Studies, Follow-Up Studies, Humans, Interferon-gamma metabolism, Interleukin-2 metabolism, Leukocytes, Mononuclear metabolism, Male, Transforming Growth Factor beta metabolism, Tumor Necrosis Factor-alpha metabolism, Bone Marrow Transplantation methods, Cytokines blood, Cytokines metabolism, Hemoglobinuria, Paroxysmal blood, Hemoglobinuria, Paroxysmal therapy
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- 2007
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17. Anti-erythroblast autoimmunity in early myelodysplastic syndromes.
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Barcellini W, Zaninoni A, Imperiali FG, Boschetti C, Colombi M, Iurlo A, and Zanella A
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- Adult, Aged, Aged, 80 and over, Anemia, Refractory therapy, Apoptosis, Bone Marrow Cells metabolism, Caspase 3 metabolism, Female, Humans, Immunoglobulin G metabolism, Leukocyte Common Antigens biosynthesis, Male, Middle Aged, Myelodysplastic Syndromes therapy, NF-kappa B metabolism, Anemia, Refractory immunology, Autoimmunity immunology, Erythroblasts metabolism, Myelodysplastic Syndromes immunology
- Abstract
Background and Objectives: Autoimmune phenomena, mainly directed against red blood cells are described in myelodysplastic syndromes (MDS), particularly early MDS, i.e. refractory anemia (RA) and RA with ringed sideroblasts (RARS). Dysregulation of apoptosis and immunoregulatory cytokines are thought to play a role in the pathogenesis of MDS., Design and Methods: This work was aimed to investigate anti-erythroid autoimmunity in unstimulated and mitogen-stimulated peripheral blood and bone marrow cultures of 26 patients with early MDS (RA and RARS), and to relate its presence with apoptotic markers and cytokine production. Bone marrow cytokine production in culture supernatants, and caspase-3 and nuclear factor-kappaB activity in cell extracts were tested by enzyme-linked immunosorbent assays., Results: Fourteen of the 26 (53.8%) patients showed the presence of autoantibodies in bone marrow cultures, whereas none displayed a positive direct antiglobulin test in peripheral blood cultures. Incubation of culture supernatants from positive patients with autologous CD45- enriched-cell suspensions showed that the autoimmune reaction was directed against autologous erythroblasts. These patients had mild signs of hemolysis and increased numbers of erythroblasts, compared with negative patients. Patients with anti-erythroblast autoimmunity displayed higher caspase-3 activity and lower tumor necrosis factor-alpha and interleukin-4 production than did negative patients., Interpretation and Conclusions: Half of the patients with early MDS showed autoimmunity against erythroblasts. This evidence might support a more rationale use of steroid therapy in these patients. The lower levels of cytokines in patients with anti-erythroblast autoimmunity are consistent with the suggested hypothesis that the autoimmune phenomena observed in MDS are probably initiated and perpetuated through alterations of pro-inflammatory and/or immunoregulatory cytokine production.
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- 2007
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18. Cytokine modulation of nuclear factor-kappaB activity in B-chronic lymphocytic leukemia.
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Zaninoni A, Imperiali FG, Pasquini C, Zanella A, and Barcellini W
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- Aged, Aged, 80 and over, Apoptosis drug effects, Case-Control Studies, DNA Fragmentation drug effects, Electrophoretic Mobility Shift Assay, Female, Humans, Interleukin-13 pharmacology, Interleukin-4 pharmacology, Leukemia, Lymphocytic, Chronic, B-Cell pathology, Leukocytes, Mononuclear drug effects, Lymphocyte Activation drug effects, Male, Middle Aged, NF-kappa B drug effects, Tetradecanoylphorbol Acetate pharmacology, Transforming Growth Factor beta pharmacology, Cytokines pharmacology, Leukemia, Lymphocytic, Chronic, B-Cell metabolism, NF-kappa B metabolism
- Abstract
Objective: Dysregulation of the apoptotic mechanisms plays a key role in the accumulation of malignant B-chronic lymphocytic leukemia (B-CLL) cells. The transcription nuclear factor (NF)-kappaB is important for cell survival by regulating the expression of anti-apoptotic genes. Several cytokines can modulate leukemic growth and apoptosis in B-CLL. The aim of this study was to determine whether cytokine-mediated regulation of apoptosis occurs via modulation of NF-kappaB activity in peripheral blood mononuclear cells from B-CLL patients., Patients and Methods: We evaluated NF-kappaB activity in peripheral blood mononuclear cells from 15 untreated B-CLL patients and 11 controls in resting conditions and in the presence of phorbol-12-myristate-13-acetate (PMA) and different cytokines by electrophoretic mobility shift assay. Apoptosis was studied by spectrophotometric analysis of DNA fragmentation., Results: We found a constitutive high NF-kappaB activity not induced by PMA in B-CLL patients, in contrast with a normal inducible NF-kappaB activity in controls. In B-CLL cultures, addition of interleukin (IL)-4 and IL-13 increased, whereas transforming growth factor (TGF)-beta reduced NF-kappaB activity compared with unstimulated cultures. Accordingly, IL-4 and IL-13 decreased, whereas TGF-beta increased DNA fragmentation compared with unstimulated cultures. IL-13 and IL-4 production was increased, whereas TGF-beta was reduced in PMA-stimulated and unstimulated cultures from B-CLL patients compared with controls., Conclusions: B-CLL patients have a constitutive high NF-kappaB activity, which is modulated by cytokines. In particular, TGF-beta displays a pro-apoptotic activity, whereas IL-4 and IL-13 have opposite effects. These cytokine alterations could be responsible for a positive autocrine circuit that maintains leukemic cells in a pre-apoptotic state., (Copyright 2003 International Society for Experimental Hematology)
- Published
- 2003
- Full Text
- View/download PDF
19. In vitro production of anti-RBC antibodies and cytokines in chronic lymphocytic leukemia.
- Author
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Barcellini W, Montesano R, Clerici G, Zaninoni A, Imperiali FG, Calori R, Cortelezzi A, and Zanella A
- Subjects
- Adult, Aged, Aged, 80 and over, Antibodies, Anti-Idiotypic analysis, Antibody Formation, Autoimmunity, Female, Humans, Immunoglobulin G immunology, Male, Middle Aged, Phytohemagglutinins, Pokeweed Mitogens pharmacology, Reference Values, Tetradecanoylphorbol Acetate pharmacology, Antibodies, Anti-Idiotypic immunology, Erythrocytes immunology, Leukemia, Lymphocytic, Chronic, B-Cell immunology
- Abstract
B-chronic lymphocytic leukemia (B-CLL) patients have a high prevalence of autoimmune phenomena, mainly autoimmune hemolytic anemia (AIHA). Immunoregulatory cytokines play a role in the regulation of both autoimmunity and leukemic B-cell growth. Mitogen-stimulated direct antiglobulin test (MS-DAT) is a recently described test able to disclose latent anti-RBC autoimmunity in AIHA. We investigated the prevalence of anti-RBC autoimmunity by MS-DAT and the pattern of cytokine production by PHA-stimulated whole blood cultures from 69 B-CLL patients and 53 controls. Results showed that anti-RBC IgG values in unstimulated, PHA-, PMA-, and PWM-stimulated cultures were significantly higher in B-CLL patients compared with controls. In B-CLL, the prevalence of anti-RBC autoimmunity was 28.9% by MS-DAT, compared with 4.3% by the standard DAT. Production of IFN-gamma, IL-2, IL-13, TNF-alpha, sCD23, and sCD30 was significantly increased in all B-CLL patients compared with controls, whereas there was no difference in IL-4, IL-6, IL-10, and TGF-beta production. Multivariate analysis showed that IL-4 was significantly increased in MS-DAT-positive compared with -negative patients. Patients with autoantibody positivity displayed greater IFN-gamma production than negative patients. These data are in line with the hypothesis that autoimmune phenomena in B-CLL are associated with an imbalance towards a Th-2-like profile. The elevated prevalence of anti-RBC autoimmunity found by MS-DAT suggests that an underestimated latent autoimmunity exists in B-CLL., (Copyright 2002 Wiley-Liss, Inc.)
- Published
- 2002
- Full Text
- View/download PDF
20. Increased Mycobacterium tuberculosis growth in HIV-1-infected human macrophages: role of tumour necrosis factor-alpha.
- Author
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Imperiali FG, Zaninoni A, La Maestra L, Tarsia P, Blasi F, and Barcellini W
- Subjects
- Chemokines biosynthesis, HIV Infections microbiology, Humans, Interferon-gamma biosynthesis, Interferon-gamma pharmacology, Interleukin-10 biosynthesis, Interleukin-10 pharmacology, Phagocytosis, Tumor Necrosis Factor-alpha pharmacology, U937 Cells, HIV-1 growth & development, Macrophages microbiology, Macrophages virology, Mycobacterium tuberculosis growth & development, Tumor Necrosis Factor-alpha biosynthesis
- Abstract
Synergism between Mycobacterium tuberculosis (M. tuberculosis) and HIV-1 infections was demonstrated in several in vitro models and clinical studies. Here, we investigated their reciprocal effects on growth in chronically HIV-1-infected promonocytic U1 cells and in acutely infected monocyte-derived macrophages (MDM). Phagocytosis of M. tuberculosis induced HIV-1 expression in U1 cells, together with increased TNF-alpha production. M. tuberculosis growth, evaluated by competitive PCR, was greater in HIV-1-infected MDM compared to uninfected cells. M. tuberculosis phagocytosis induced greater TNF-alpha and IL-10 production in HIV-1-infected MDM than in uninfected cells. In uninfected MDM, addition of TNF-alpha and IFN-gamma decreased, whereas IL-10 increased M. tuberculosis growth. On the contrary, in HIV-1-infected MDM, addition of TNF-alpha and IFN-gamma increased, whereas IL-10 has no effect on M. tuberculosis growth. TNF-alpha seems to play a pivotal role in the enhanced M. tuberculosis growth observed in HIV-1-infected MDM, being unable to exert its physiological antimycobacterial activity. Here, for the first time we demonstrated an enhanced M. tuberculosis growth in HIV-1-infected MDM, in line with the observed clinical synergism between the two infections.
- Published
- 2001
- Full Text
- View/download PDF
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