Giuseppe Iodice

Detection of hepatitis C virus (HCV) proteins by immunofluorescence and HCV RNA genomic sequences by non-isotopic in situ hybridization in bone marrow and peripheral blood mononuclear cells of chronically HCV-infected patients

Immunofluorescence (IF) to detect HCV antigens and non‐isotopic in situ hybridization (NISH) to detect HCV RNA genome were carried out on bone marrow (BM) and peripheral blood (PB) mononuclear cells (MC) of 11 chronically HCV‐infected patients. In four patients (36.4%) HCV antigens were detected in monocytes macrophages as well as in B lymphocytes in both BMMC and PBMC. Positive T lymphocytes in BMMC were found in three of them, but only one patient showed positive T cells in PBMC. NISH invariably demonstrated minus and plus HCV RNA genomic strands either in monocytes macrophages or B and T lymphocytes in BMMC and PBMC in the four HCV antigen‐positive patients and in two further patients not expressing viral proteins in blood MC. IF signals appeared diffusely distributed within the cytoplasm, or as brilliant granules in distinct submembrane areas or else in cytoplasm membrane. Nuclei never stained. Similarly, NISH displayed HCV RNA accumulation restricted to MC cytoplasm only, nuclei being persistently negative. NISH, however, was unable to detect cell membrane signal. Infection of blood MC is a common event in naturally acquired HCV infection, since none of these patients was conditioned by immunomodulating or immunosuppressive therapies. No difference was found in terms of mean age, length of disease, anti‐HCV immune response, type and severity of chronic liver damage between patients with HCV‐infected MC and patients without cell infection. These results demonstrate that HCV can infect BMMC and PBMC that represent important extrahepatic sites of virus replication, and may help to explain the immunological abnormalities observed in chronic HCV carriers.

Treatment of adult patients with idiopathic thrombocytopenic purpura with intravenous immunoglobulin: effects on circulating T cell subsets and PWM-induced antibody synthesis in vitro

Summary. Eight adult patients with idiopathic thrombocytopenic purpura (ITP) were given two 5‐d courses of intermediate (250 mg/kg body weight/d) to high dose (400 mg/kg body weight/d) intravenous infusions of human plasmin‐cleaved Ig, at 15–30 d intervals. Three patients also were given single booster infusions (400 mg/kg body weight for 1 d) of a different preparation, S‐sulfonated Ig. As expected, significant though transient rises in the platelet count were consistently observed in all patients. The mean platelet count increase was 95 600/mm3 after the first course, and 143 500/mm3 after the second course. Similar effects of lower magnitude were obtained several times in the patients given single booster doses. In three patients, platelet‐bound IgG levels were decreased in association with Ig therapy.

High-dose thiotepa, etoposide and carboplatin as conditioning regimen for autologous stem cell transplantation in patients with high-risk Hodgkin’s lymphoma

Abstract Background: Autologous stem cell transplantation (ASCT) generally provides good results in Hodgkin’s lymphoma (HL). We studied a high-dose chemotherapy regimen based on thiotepa, etoposide and carboplatin (TECA). Methods: Fifty-eight patients with advanced HL were treated with thiotepa, etoposide and carboplatin for transplant induction. Results: The overall response rate was 79·3% (39 CR: 67·2%; and 7 PR: 12·1%); 12 patients (20·1%) were non-responders. The 5-year overall survival rate was 77·6%; five initially responder patients relapsed within the first 5 years of follow-up and underwent salvage therapy. Conclusion: The TECA conditioning regimen for ASCT in HL results in a good anti-HL effect, positive response to treatment and high 5-year overall survival rate. It was also well tolerated and did not induce excessive toxicity, suggesting that TECA may be a very useful conditioning regimen for HL.

Induction Therapy and Stem Cell Mobilization in Patients with Newly Diagnosed Multiple Myeloma

Autologous stem cell transplantation (ASCT) is considered the standard therapy for younger patients with newly diagnosed symptomatic multiple myeloma (MM). The introduction into clinical practice of novel agents, such as the proteasome inhibitor bortezomib and the immunomodulatory derivatives (IMiDs) thalidomide and lenalidomide, has significantly contributed to major advances in MM therapy and prognosis. These novel agents are incorporated into induction regimens to enhance the depth of response before ASCT and further improve post-ASCT outcomes. Between January 2000 and November 2011, 65 patients with MM were transplanted in the Department of Biomedical Science and Clinical Oncology at the University of Bari. According to Durie-Salmon, 60 patients had stage III of disease and 5 stage II. Only 7 patients were in stage B (renal failure). Induction regimens that were administered in two or more cycles were VAD (vincristine, adriamycin, and dexamethasone), Thal-Dex (thalidomide, dexamethasone), Len-Dex (lenalidomide, dexamethasone), Vel-Dex (bortezomib, dexamethasone), VTD (bortezomib, thalidomide, and dexamethasone), and PAD (bortezomib, pegylated liposomal doxorubicin, and dexamethasone). In mobilization procedure, the patients received cyclophosphamide and granulocyte colony-stimulating factor (G-CSF). The number of cells collected through two or more leukapheresess, response after induction, and toxicity were evaluated to define the more adequate up-front induction regimen in transplantation-eligible MM patients.

90Y-Ibritumomab Tiuxetan as Consolidation Therapy After Autologous Stem Cell Transplantation in Aggressive Non-Hodgkin Lymphoma

Targeted radioimmunotherapy with 90Y-labeled ibritumomab tiuxetan is a novel therapeutic approach for CD20-positive relapsed or refractory non-Hodgkin lymphoma (NHL). Methods: Seven consecutive patients with CD20-positive aggressive NHL who did not fully respond to prior myeloablative chemotherapy were enrolled. A 14.8 MBq (0.4 mCi)/kg dose of 90Y-ibritumomab tiuxetan was administered to all patients, and approximately 100 d afterward 18F-FDG PET/CT was performed to assess response. Results: PET/CT showed a complete response in 5 of 7 patients. Of the 2 nonresponsive patients, 1 showed persistent disease and the other progression. Toxicity included thrombocytopenia in all 7 patients and grade IV neutropenic fever in 1 patient. Conclusion: Despite the small series studied, we suggest that radioimmunotherapy is safe for consolidation in patients treated with high-dose chemotherapy for aggressive NHL and may provide clinical benefit in extensively pretreated patients.

Analogues in immunology.

SummaryA number of immunological functions has been found to be endowed with biological rhythmicity. Variations of peripheral blood lymphocytes, immediate and delayed hypersensitivity responses, and reactivity of the immune system to antigenic challenge have consistently shown circadian bioperiodicity. By employing a panel of monoclonal antibodies specific for the various lymphocyte subsets, we have been able to detect rhythmic variations in the number of total T lymphocytes, as well as of the T helper (TH) and the T suppressor/cytotoxic (Ts) subsets. Available evidence clearly indicates such circadian variations to be due to compartmentalization of circulating lymphocytes in several lymphoid organs. Biological rhythms (circadian, circaseptan, etc.) have been demonstrated in several immunological situations of clinical interest, including variations of TH and Ts cells in patients with rheumatoid arthritis incidence of allograft rejections, antitumoral effectiveness of chemoimmunotherapy, and allergic symptoms. In addition, the recent availability of synthetic analogues of some thymic peptide hormones and of the synthetic ACTH 1–17 (Synchrodyn® 1–17 is greatly contributing to the elucidation of the role of such hormones in the immune regulation process. A preliminary account is reported of the experiments performed with the aim of raising conventional rabbit antisera to the peptide analogue ACTH 1–17, as well as of the consistently negative results which were obtained when 578 human serum samples (from patients treated with such synthetic peptide) were screened by a radioimmunoassay for the occurrence of anti-ACTH 1–17 antibodies. It is emphasized that the recognition of the rhythmic variations of several immunological mechanisms and of the crucial role played by the analogues of adrenal corticotropic and thymic hormones in immune regulation has favoured significant advances in the fast growing area of chronoimmunology.