Atherosclerosis and inflammation. Patterns of cytokine regulation in patients with peripheral arterial disease.

Inflammatory phenomena at sites of atherosclerotic plaques are increasingly thought to be major determinants of the progression and clinical outcome of atherosclerotic disease. Therefore, attention is being paid to systemic markers/mediators which may reflect the inflammatory activity in the plaques. This study evaluates the pattern of the main proinflammatory cytokines tumor necrosis factor-alpha (TNFalpha), interleukin-1beta (IL-1beta), and interleukin-6 (IL-6), their soluble receptors/antagonist, and a variety of inflammatory markers, in patients with peripheral arterial disease (PAD). Eight patients with PAD suffering from claudicatio intermittens (CI), eight with critical limb ischemia (CLI) and eight controls (C) were studied. Blood samples were collected at baseline in all groups and. for C and CI, immediately after and 4 h after a 30-min treadmill test. Baseline: no differences in cytokine plasma levels were detected among the three groups. In contrast, soluble receptors of TNF (type I and II) and of IL-6, and IL-1beta receptor antagonist (IL-1ra) were increased in CI and CLI patients, as compared to C. Of note, IL-Ira correlated with the occurrence and stage of the disease in a highly significant proportion of the patients, reaching a predictive value for the disease of P < 0.0001. The opposite trend was observed for the soluble receptor of IL-1beta. Notably, in the patients no alterations could be found in white blood cell counts, expression of CD11c adherence molecule by circulating monocytes or, in vitro. O2- release from zymosan-activated neutrophils. Moreover, plasma levels of platelet activating factor (PAF), of neutrophil elastase and of the acute phase reactants C-reactive protein (CRP) and alpha1-acid glycoprotein were not found to be significantly altered. In contrast, the acute-phase proteins alpha1-antitrypsin (alpha1AT) and haptoglobin (HG) were found to be increased. Effect of treadmill: IL-1beta and TNFalpha remained at baseline levels following exercise, and IL-6 dropped to undetectable levels. Among cytokine antagonists, again the most relevant changes concerned the IL-1ra, which was significantly increased immediately after the treadmill test, both in CI and C, and returned to baseline levels after 4 h. In contrast, soluble TNFalpha, IL-1beta and IL-6 receptors, PAF, and the other markers of leukocyte activation were not found to be altered. Soluble TNFalpha and IL-6 receptors were shown to inhibit the biological effects of their ligands. Similarly, IL-1ra and the acute phase proteins alpha1AT and HG have been reported to exert anti-inflammatory functions. The increased plasma levels of these agents, together with low levels of inflammatory cytokines and other pro-inflammatory mediators such as PAF and alpha1-acid glycoprotein, appear to draw an undescribed picture, so far, of upregulation of a composite systemic anti-inflammatory mechanism in atherosclerotic patients. IL-1ra appears to be a reliable marker of the state of activation of this mechanism. These results may provide a basis for developing new insights into the pathogenesis of the atherosclerotic disease.

Immunoreactivity of chimeric proteins carrying the HIV-1 epitope IGPGRAF. Correlation between predicted conformation and antigenicity.

Sera from HIV-1 infected individuals were examined for their reactivity to the principal neutralizing domain, IGPGRAF sequence, of the V3-loop of HIV-1. Four hybrid proteins carrying this sequence inserted in four different outer loops of a protein that makes up the capsid of an insect virus were used as antigen in a Western blot assay for this survey. All the four antigens showed different activity: sera that recognise all antigens to sera that reacted with only one of them. Competition experiments indicated that the antibodies recognised these proteins with different affinity. Molecular modelling of the hybrid proteins predicted that the inserted sequence adopted different conformations in each position. Comparison of predicted most stable conformations for IGPGRAF indicated that there is a close relationship between conformational similarity to a V3-loop reference structure and the degree of reactivity with sera.

Low-dose "natural" alpha-interferon in B-cell derived chronic lymphocytic leukemia.

BACKGROUND: Alpha-interferon (alpha-IFN) was found to have a good antiproliferative effect in early stage chronic lymphocytic leukemia (CLL), but recombinant alpha-IFN administration may induce serious side effects. Therefore low-dose "natural" IFN was evaluated in terms of efficacy and safety. METHODS: Fifteen patients affected by stage A (according to Binet) B-CLL underwent the treatment: natural IFN 1 MU three times a week for 6 months. RESULTS: Overall lymphocyte count decreased from 13,050 +/- 3,200 to 7,500 +/- 2,940 within 6 months. One patient did not respond to IFN therapy. No one complained of side effects. CONCLUSION: Low dose "natural" alpha-IFN seems useful and well tolerated in CLL, but the potential curative role of IFN in CLL remains to be established.

Mechanisms of eosinophil adherence to cultured vascular endothelial cells. Eosinophils bind to the cytokine-induced ligand vascular cell adhesion molecule-1 via the very late activation antigen-4 integrin receptor.

We have examined the mechanisms involved in the adherence of normal peripheral blood eosinophils to cultured human umbilical vein endothelial cells (HEC) under three conditions: (a) adherence in the absence of treatment of HEC or eosinophils with activating agents (basal adherence); (b) adherence induced by stimulation of eosinophils with phorbol ester (eosinophil-dependent adherence); and (c) adherence induced by pretreatment of HEC with LPS, tumor necrosis factor (TNF), or IL-1 (endothelial-dependent adherence). A mechanism was identified that was equally active in basal, eosinophil-dependent, and endothelial-dependent adherence. This mechanism was optimally active in the presence of both Ca++ and Mg++, and reduced in the presence of Ca++ only or Mg++ only. Furthermore, like the other mechanisms of eosinophil adherence, it was active at 37 degrees C but not at 4 degrees C. A second mechanism of adherence was involved in eosinophil- and in endothelial-dependent adherence. This mechanism was dependent on the CD11/CD18 adhesion complex of eosinophils (i.e., inhibited by anti-CD18 MAb) and it was active in the presence of Ca++ and Mg++ or Mg++ only, but not Ca++ only. The third mechanism of adherence was specific for endothelial-dependent adherence. It involved the endothelial ligand vascular cell adhesion molecule-1 (VCAM-1) and the eosinophil receptor very late activation antigen-4 (VLA-4, CD49d/CD29, i.e., inhibited by anti-VCAM-1 MAb or anti-VLA-4 MAb). This mechanism was active in the presence of Ca++ and Mg++ but not of Ca++ only or Mg++ only, and was not up- or downregulated when eosinophils were stimulated with phorbol ester. In contrast, the endothelial leukocyte adhesion molecule-1 (ELAM-1), that binds neutrophils and monocytes, was not involved in eosinophil adherence to LPS-, TNF-, or IL-1-stimulated HEC (i.e., not inhibited by anti-ELAM-1 MAb). We conclude that eosinophils, like monocytes and lymphocytes, bind to the cytokine-induced endothelial ligand VCAM-1 via the integrin receptor VLA-4.

[Chronic granular T lymphocytosis of undetermined significance]. / T linfocitosi granulare cronica di significato indeterminato.

The clinical, morphologic, immunologic, functional, serologic and cytogenetic features of 4 cases of chronic granular T cell lymphocytosis with neutropenia were studied. The records of these patients were reviewed and an infectious event preceding the development of the disorder could be documented in 3 cases, suggesting a cause and effect relationship. A benign monoclonal gammopathy was detected in the remaining patient. The clinical picture was characterized by moderate blood and bone marrow lymphocytosis, neutropenia, hepatosplenomegaly, absence of lymphadenopathy, and a stable course, observed over a period of up to 11 years. Surface marker analysis in all the patients showed the common membrane phenotype of granular T cell lymphocytosis (CD3+, CD4-, CD8+, Leu7+). One patient treated with steroid therapy had reversal of lymphocytosis and severe neutropenia, which both recurred after steroids were withdrawn. The disorder again resolved after pulse steroid treatment. From these findings and a review of the literature, we suggest that granular T cell lymphocytosis with the immunologic phenotype exemplified by our cases is a distinct reactive or immunoregulatory disorder. In view of the similarities in character to benign monoclonal B cell lymphocytosis and idiopathic paraproteinaemia, we suggest that this disorder be termed chronic granular T cell lymphocytosis of undetermined significance.

HLA-B39 and the axial type of psoriatic arthritis.

The frequency distribution of HLA-B38 and HLA-B39 was studied in a group of 50 unrelated psoriatic arthritis patients. A high increase of HLA-B39 was noted in the subgroup with axial arthropathy. All B39 positive patients presented with spondylitis and/or sacroiliitis. The hypothesis is formulated that HLA-B39 may discriminate a genetic subset of psoriatic arthritis characterized by a particular susceptibility to develop axial involvement.

HLA antigens in familial hyper-alphalipoproteinemia.

Tissue typing for HLA-A, B and C antigens was performed in 28 subjects belonging to a family in which an inherited increase of high density lipoprotein cholesterol (HDL-C) is present. Nine subjects had definite hyper-alphalipoproteinemia (HDL-C greater than 80 mg/dl) and 3 "borderline" values (HDL-C greater than 70 mg/dl). There is not any linkage between the presence of HDL-C elevation and the HLA haplotypes.

Association between HLA-B16 and psoriatic spondylitis.

The frequency distribution of HLA antigens was studied in a group of 40 patients with psoriatic arthritis from a region in North-eastern Italy. Our results indicate: 1) a strong association, in this population, between HLA-B16 and inflammatory spinal involvement; 2) a failure to confirm previous reports on HLA-B27 increased frequency in psoriatic spondylitis and/or sacroiliitis.

Incidence of paraproteinaemia in blood donors.

Paraproteinemia was detected by agarose gel electrophoresis in 13 out of 3,800 apparently healthy blood donors of an Italian district. None of the subjects with a serum M component developed overt B cell malignancy over a prolonged period of observation. The incidence of paraproteinaemia was higher than that reported from previous surveys of similar populations of blood donors in other European countries, possibly due to the sensitivity of the technique adopted for the screening investigation. The main implications of the detection of monoclonal immunoglobulin abnormalities are briefly discussed with particular reference to the possible indications for routine screening of blood donors for the presence of paraproteins.

Auto-reaction phenomenon detected by enzyme electrosyneresis in the serum of patients with viral hepatitis and other diseases with immune complexes.

The use of modified electrosyneresis by making 760 sera of healthy persons or persons suffering from various diseases with immune complexes to react with their own pronase-treated serum has shown the following results: - One of 220 sera of healthy persons, 11 were positive in ESE (5%); - Out of 123 sera of HBsAg carriers, 23 were positive (18.6%); - Out of 135 sera of patients with acute viral type B hepatitis, 132 were positive (97.7%); - Out of 168 sera of patients with acute HBsAg negative hepatitis, 127 were positive (75.5%); - 4 cases of fulminant hepatitis were all strongly positive; - 54 cases of patients with rheumatoid arthritis were 100% positive; - 2 cases of patients with systemic lupus erythematosus were positive; - Out of 6 patients with glomerulonephritis 3 were positive; - Out of 34 patients with carcinoma of various organs, 19 were positive (55.88%). The Authors believe, on the basis of the research work set forth partly in the present report, that this interaction is connected with the presence of immune complexes. The method used has been called Enzyme Electrosyneresis (ESE) and the detected antigen "Auto antigen exposed by protease and by electrosyneresis" (AEPE).

[Detection of paraproteinemia in blood donors. Results of systematic analysis of 3800 donors in Trieste]. / La ricerca della paraproteinemia nei donatori di sangue. Risultati dell'analisi sistematica di 3800 donatori della provincia di Trieste.

A systematic analysis of plasma protein pattern performed by agarose gel electrophoresis has detected paraproteinemia in 13 out of 3800 blood donors in the district of Trieste. The variations in the incidence of monoclonal gammopathies in surveys of blood donors could be related not only with geographical origin and the age and sex distribution of the population studied but also with the sensitivity of the screening technique employed. The physiopathological and clinical implications of the detection of paraproteins favour the inclusion of the screening plasma electrophoretic analysis among the laboratory investigations for blood donors.

[HBsAg and HBsAb in carriers and their families]. / HBsAg ed HBsAb in portatori e loro familiari

Relatives of 98 Au antigen carriers were examined (positivity by AUSRIA II-125), already donors at the Immunotransfusion Centre of Trieste General Hospitals, totalling 106 cases: 98 out of same were represented by their wives and children, older than 7 years. 8 of the wives were HB,sAg (8.2%) positive and 90 negative: children were all antigen negative. In wives and children the presence of eventual anti-HBsAg antibodies by means of AUSAb technique (Abbott) was investigated obtaining 29 positive (29.3%) and 69 negative sera. With Dr. Lacy's (Abbott Diagnostic Division) collaboration the typing of HBsAg and HBsAb found was performed in order to obtaine epidemiological informations.