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Background and aims Dyslipidaemias are associated with cardiovascular mortality and morbidity, driven by unstable atherosclerotic plaques with inflammatory infiltrates. Levels of messenger RNA (mRNA) for pro-inflammatory cytokines have been positively correlated with atherosclerotic disease progression. Therapeutic lipoprotein apheresis (LA) reduces plasma lipid levels and reduces inflammation. We evaluated the effects of LA on expression of mRNA coding for key pro-inflammatory cytokines in patients with dyslipidaemia, homo-/hetero-zygous familial hypercholesterolaemia (HoFH, HeFH) or hyperlipoprotein(a)aemia [hyperLp(a)] and associated coronary artery disease (CAD). Approach Ten patients (five males and five females, mean age 47 ± 9.2 years) were enrolled, all with HyperLp(a) or confirmed genetic diagnoses of dyslipidaemia, HoFH, or HeFH; all had associated CAD. mRNA determinations were via reverse transcriptase polymer chain reaction (RT-qPCR). Results LA was associated with downregulation of mRNA expression for IL-1α, IL-6 and TNF-α, starting after the first LA session. The observed reduction was progressively enhanced during the interval between the first and second LA sessions to achieve a maximum decrease by the end of the second session (IL-1α: −49%, p Conclusions LA suppresses the expression of IL-1α, IL-6 and TNF-α mRNA in patients with dyslipidaemias. This may contribute to the arterial anti-inflammatory effect of LA.

The effect of lipoprotein apheresis (Direct Adsorption of Lipids, DALI) (LA) on plasma levels of pentraxin 3 (PTX3), an inflammatory marker that reflects coronary plaque vulnerability, and expression of PTX3 mRNA was evaluated in patients with hyperLp(a)lipoproteinemia and angiographically defined atherosclerosis/coronary artery disease. Eleven patients, aged55±9.3years (mean ± SD), were enrolled in the study. PTX3 soluble protein levels in plasma were unchanged by 2 sessions of LA; however, a downregulation of mRNA expression for PTX3 was observed, starting with the first session of LA (p<0.001). The observed reduction was progressively increased in the interval between the first and second LA sessions to achieve a maximum decrease by the end of the second session. A statistically significantly greater treatment-effect correlation was observed in patients undergoing weekly treatments, compared with those undergoing treatment every 15 days. A progressive reduction in plasma levels of C-reactive protein was also seen from the first session of LA, with a statistically significant linear correlation for treatment-effect in the change in plasma levels of this established inflammatory marker (R2=0.99;p<0.001). Our findings suggest that LA has anti-inflammatory and endothelium protective effects beyond its well-established efficacy in lowering apoB100-containing lipoproteins.

We have used genomic probes which specifically recognize DNA rearrangements of the RAR-alpha locus on chromosome 17q21 in patients with acute promyelocytic leukaemia (APL) and acute myeloid leukaemia (AML) subtypes. Molecular data were examined in comparison with morphological and immunophenotypic characterization at diagnosis in 20 hypergranular FAB M3 cases, five microgranular APL (M3v), 51 non-M3 AML and 12 myeloid CML blast crises. Rearrangements of the RAR-alpha locus were only detected in 23/25 APL cases and in none of the other FAB subtypes analysed. Surface marker characterization showed a consistent immunophenotypic profile--HLADR negative, CD9 and CD13/33 positive--in all M3 and M3v cases. Neither HLADR negativity nor CD9 positivity were associated with RAR-alpha rearrangements in non M3 AML. Our data indicate that RAR-alpha gene rearrangements are relevant diagnostic features of both M3 and M3v, and may prove useful molecular marker for follow-up analysis in APL patients.

We report the results of a randomized clinical trial of two different coricosteroids (prednisone versus deflazacort) in patients affected by autoimmune thrombocytopenic purpura (ATP). We have evaluated the efficacy of the two steroids on platelet count, antiplatelet antibodies, lymphocyte subsets and the occurrence of side effects. Twenty-seven patients were evaluable: 13 were treated with PDN and 14 with DFC. After 24 weeks of treatment, 4/12 (33%), subjects treated with PDN were refractory while complete responses were obtained in 2/12 (17%) and partial responses in 6/12 (50%). Among patients treated with DFC, 4/11 (36%) were considered as refractory, 2/11 (18%) had a complete response and 5/11 (46%) a partial response. A statistically significant decrease of antiplatelet antibodies was recorded in both groups after 4 weeks of therapy, but only in subjects receiving PDN did the reduction last until the 24th week. We observed an increase of T lymphocyte subsets (CD3, CD2, CD4, CD8) in absolute number, due to an increase in circulating lymphocytes, after 4 weeks. No substantial modifications were observed in these populations regarding the percentage or the CD4/CD8 ratio. After 24 weeks, 91% (10/11) of patients treated with PDN presented an increase of body weight and 1 had a stable increase in blood pressure. Among the subjects treated with DFC, 64% (7/11) showed an increase of body weight after the same follow-up. In conclusion, no difference was observed the two steroids studied.

Acute promyelocytic leukemia (FAB-M3) is a distinct entity among acute non-lymphoid leukemias (ANLL) with peculiar morphological, biological, clinical and prognostic features. An atypical form of M3 (M3v) could be confused with other FAB ANLL and therefore the diagnosis of this variant requires ultrastructural analysis and/or cytogenetic study and/or selective gene rearrangement studies. The immunological phenotype of blast cells in 39 APL patients was studied at diagnosis. The diagnosis of M3 FAB type was ascertained in 32 and the diagnosis of M3v in 7 cases. Using a large series of monoclonal antibodies (mAb), the APL blast cells were B and T cell antigens-negative, HLA-DR constantly negative, CD13- and/or CD33-positive, CD9-positive. Among ANLL this phenotype seems to be closely related to APL both in M3 type and M3v subtype. Because the diagnosis of APL (M3 or M3v) is important in order to establish the specific therapeutic approach, the discriminant capacity of the immunological typing to identify M3 and mainly M3v (hypogranular) could be determinant for a "quick" diagnosis.

The expression of complement receptors (C3R) associated or not to sheep erythrocyte receptors (ER) and surface membrane immunoglobulins (SmIg) was determined on lymphoblasts of 45 patients with acute lymphocytic leukemia (ALL) at onset and/or in the first relapse. We found C3R simultaneously expressed with ER or SmIg on lymphoblasts of T cell ALL and B cell ALL. Lymphoblasts were positive for C3R in absence of ER and SmIg only in three patients in the hematological relapse. We stress the importance to find C3R as independent marker on ALL cells at onset and in subsequent relapses.

The configuration of the immunoglobulin heavy chain (IgH), T-cell receptor (TcR) beta and gamma chain regions, and the major breakpoint cluster region (M-bcr) genes were analysed in four cases of Ph' + acute leukemia (AL). Monoclonal rearrangements of the IgH region were detected in three cases exhibiting two phenotypically distinct cell populations (i.e. one lymphoid and one myeloid. In one of these cases, identical genetic events were observed by molecular analysis of FACS separated blasts. Multi-lineage rearrangements involving also the TcR gamma gene were observed in a biphenotypic AL showing co-expression of markers. The lack of rearrangements within the M-bcr gene, together with demonstration in one case of the Ph' + AL specific p190 protein product, pointed against the occurrence of chronic myeloid leukemias presenting in blastic transformation. Our results imply that such cases are to be considered as true AL and should therefore be included in the definition of hybrid AL.

In the present report we describe the immunological features of peripheral blood mononuclear cells (PBMC) isolated from two patients with hematological disorders. The majority of cells from both patients, although capable of rosetting with sheep erythrocytes, lacked T cell-specific or myelomonocytic-specific antigens, as detected by a large panel of monoclonal antibodies. High proportions of PBMC from both patients had receptors for the Fc portion of IgG and were able to strongly exert cytotoxic/suppressor activities. These cells, showing some distinctive morphological features (namely the presence of granules in their cytoplasm) at the electron microscope level, appear to represent a discrete subset of PBMC, whose lineage, presence in normal donors, and functional properties are still to be defined.

Between January 1984 to June 1985, 18 Ph1 positive chronic myeloid leukemia (CML) patients in chronic phase (CP) underwent allogeneic bone marrow transplantation (BMT) from HLA identical and MLC negative siblings. The median age was 32.5 yr and median disease duration of CML at time of BMT was 19.3 months. The pretransplant conditioning regimen consisted of cyclophosphamide (CTX) (120 mg/kg) and 10.20 Gy total body irradiation (TBI) at 6 doses of 1.7 Gy each, administered in 3 daily fractions over 2 days at a dose rate of 15-20 cGy/min. To prevent graft-vs-host disease (GvHD) we used methotrexate (MTX) in one patient and cyclosporin-A (CYA) in the other 17 patients. In addition to CYA, given until day +365, 10 patients received donor marrow depleted of T cells with CAMPATH-1. The residual marrow lymphocytes were always less than 1%. The rate of engraftment was significantly correlated with the number of nucleated cells infused. Neither GvHD nor graft failure were observed among CAMPATH-1 patients. In this group one cytogenetic and one hematologic relapse occurred. The overall actuarial survival at 24 months is 78%. Of the 10 patients treated with donor marrow depleted of T cells, 9 are alive after a median follow-up of 9 months (range 5-18), with an actuarial survival of 90%. Of the other 8 patients transplanted with untreated marrow, 5 are alive after a median follow-up of 19.3 months (range 3.7-24) and the actuarial survival is 63.8%. This pilot study seems to demonstrate that T-cell depletion of donor bone marrow with CAMPATH-1 is effective to prevent GvHD, while the risk of graft failure can be avoided using a "standard" conditioning regimen including a fractionated TBI with a fast dose rate and a prolonged administration of CYA at the maximum tolerable dosage. While the high frequency of relapses suggests the employ of more aggressive anti-leukemic conditioning regimens in CAMPATH-1 treated marrow recipients.

Three hemophiliacs with high titre inhibitor were treated with a medium-high FVIII dose schedule (100 IU/kg bw daily) with the aim of inducing the immunotolerance. These patients were followed-up extensively concerning their immunological status and HIV serology. In all of them the inhibitor disappeared and normal FVIII kinetics were obtained after 22, 15 and 29 months. After eradication of the inhibitor, no recurrence took place in any of the patients. All the patients were HIV Ab positive before the beginning of the treatment. In one of them CD4+ cells fell progressively 32 months after the treatment was started, a full-blown AIDS showed up, and the patient died 5 1/2 years after the beginning of the treatment. In the second and third patient the CD4+ cells varied widely but remained greater than 400/microliter during the whole immunotolerance treatment. The latter two patients are AIDS and ARC free so far, but patient No. 2 developed a mild-to-severe thrombocytopenia. Considering the high cost of the treatment and the possibility that such an intensive administration of FVIII concentrates might worsen the immunological status of patients, this therapeutic procedure should only be applied with caution.

An assessment of the lymphocyte sub-populations and cell mediated cytotoxicity was made in 12 adults with acute non-lymphoid leukemia after stopping chemotherapy. No significant differences were found between ANLL and controls, for absolute lymphocyte count, %SIg/+ cells, Fc gamma + cells and ERFC. The OKT4+/OKT8+ was reduced in ANLL. Leu-7+ cells were significantly higher in ANLL. Moreover it appears that in ANLL lymphocyte function is restored early after stopping chemotherapy.

Two childhood and three adult patients with acute lymphoblastic leukemia (ALL) are described. The identification of the T-cell nature of their ALL-cells was based on the positivity of their blasts with 3A1 monoclonal antibody which recognizes immature and mature T-cells. Four tested cases showed a terminal deoxynucleotidyl transferase (TdT) positivity, and four out of five were acid phosphatase (AcP)-positive. Cells from all cases were characterized with a panel of 15 monoclonal antibodies: 3A1 was constantly positive; J5, BA-1, OKT6, Leu-7, and other monoclonal antibodies reactive with T-cells (OKT3, OKT4, OKT8, OKT11, Leu-1, T65) were negative; only two cases had weak positivity with OKT11 and Leu-1, respectively; cells from all cases were surface immunoglobulin (SIg)-negative; three out of the five cases were OKT10-positive and three OKT9-positive; none of the cases reacted with OKM1 and anti-HLA-DR. Our findings indicate that the use of 3A1 monoclonal antibody was helpful in the recognition of individual cases of T-ALL otherwise considered as unclassified-ALL (U-ALL). 3A1 and an anti-common ALL Antigen (CALLA) reagent may represent the essential monoclonal antibodies to be used for screening ALL cells.

The immunophenotype of 135 previously untreated patients with FAB defined acute myeloid leukaemia (AML) was studied at diagnosis. The panel of reagents included monoclonal antibodies (MoAb) recognising myeloid-associated determinants (CD11, CD13, CD14, CD33 and others) as well as MoAb directed towards lymphoid antigens (CD7, CD10, CD19) and TdT. The results indicate that CD13 and/or CD33 are consistently expressed in AML and only rarely in ALL blasts (131/135 + ve cases, versus 4/130 in ALL). Lymphoid antigen expression was rarely detected when CD10 and CD19 were investigated in AML (0.9% and 2% + ve cases, respectively), whereas significant positivities were found for TdT and CD7 (20% and 10% respectively). Concerning FAB subtypes, two new MoAb (LAM3 and LAM7) proved very useful in the specific recognition of AML with monocytic features. The phenotype CD13+ and/or CD33+, CD9+, HLA-DR- was found to be almost exclusive for M3 AML. The response to induction chemotherapy was analysed in CD7+ and in TdT+ patients. In the latter group a statistically significant lower response rate was found with respect to TdT-ve-AML patients.

A quantitative evaluation of terminal deoxynucleotidyl transferase (TdT) was performed using a highly sensitive enzyme immunoassay (EIA) in 72 previously untreated patients with acute myeloid leukemia (AML). Biological analysis of the leukemic cells included in all cases cytochemistry, search for Ph' chromosome and immunophenotyping with both anti-lymphoid and anti-myeloid monoclonal antibodies (MoAbs). Thirteen AML cases (18 per cent) were considered TdT+ by EIA. According to the FAB classification, almost all of them (12 out of 13) were within the M1 and M2 subgroups. A mixed lymphoid-myeloid phenotype was observed in one of the 13 TdT+ cases, while in none of the others were lymphoid features detected. Nine of the 10 EIA TdT+ cases studied in parallel were TdT positive with the conventional immunofluorescence assay. All patients received standard protocol chemotherapy and in 61 (13 TdT+, 48 TdT--) the response to induction treatment was analysable. Only 3/13 TdT+ patients (23 per cent) achieved a complete remission (CR), while in the TdT- group 38 patients had a CR (79 per cent) and 10 were resistant (p less than 0.01). It is suggested that the incidence, biological interest and prognostic significance of TdT+ AML should encourage the routine and more accurate search for this marker in all patients with AML.

A multiparameter analysis, which included the evaluation of clinical features, cell morphology, karyotype, phenotypic and functional immunologic findings, and T-cell receptor beta-chain configuration was performed on 34 patients with lymphoproliferative disease of granular lymphocytes (LDGL). The two-fold aim of the study was to identify the most useful tools that would more accurately characterize these patients and to deal with the problem of classifying these lymphoproliferative disorders. The data presented in this article suggest that a single parameter may not be sufficient to define the nature of the proliferating cells or to predict the clinical course of the disease and prognosis for the patient. The use of a multiparameter approach, however, may reach this goal, thus providing important prognostic and therapeutic information. Our study supports the concept that lymphoproliferative disease of granular lymphocytes is a heterogeneous disorder that ranges from indolent and possibly reactive conditions to the manifestation of aggressive malignancies.