Peripheral blood mononuclear cells of patients with systemic sclerosis produce increased amounts of interleukin 6, but not transforming growth factor beta 1.

OBJECTIVE: To assess the ability of peripheral blood mononuclear cells (PBMC) of patients with systemic sclerosis (SSc) to produce interleukin 6 (IL-6), transforming growth factor beta 1 (TGF-beta 1), to identify the IL-6 producer cells in the in vitro model, and to correlate these data with the clinical evidence of our patients. METHODS: We used a sandwich ELISA to quantitate IL-6 and TGF-beta 1 levels in sera, plasma, and supernatants, and an imunofluorescence technique to evaluate IL-6 producing cells in our patients. RESULTS: IL-6 was detected in sera from 8 of 20 patients and no controls (p < 0.05). A significant increase of IL-6 production was observed in both spontaneous and phytohemagglutinin (PHA) induced cultures of PBMC from patients with SSc vs controls. No differences in TGF-beta 1 production were observed, either in sera or supernatants, between patients and controls. A significant increase of IL-6 synthesizing cells was observed after 3 h of PHA stimulation in patients vs controls (p < 0.05). CONCLUSION: Spontaneous IL-6 production and the higher number of IL-6 producing cells in patients with SSc suggest that these cells have been already primed in vivo. The absence of PBMC primed for TGF-beta 1 production supports the hypothesis that cells other than lymphocytes produce and secrete this cytokine in the skin of patients. Higher serum levels of IL-6 observed in a subset of patients did not correlate with either severity or duration of disease.

[Systemic sclerosis after a long-term follow-up]. / Riflessioni sulla sclerosi sistemica dopo un follow-up a lungo termine.

The authors suggest some remarks on the aetiopathogenesis of systemic sclerosis through a critical review of a group of 51 patients affected by this disease for many years. In this paper the authors report their previous studies regarding the correlation of immunological response, both cell-mediated and humoral, hemocoagulative aspects and haemorheological changes. From the results of immunological studies, the authors report a long-term treatment with an immunomodulating drug: thymopentin. Moreover they report a recent work about the connection among stressor event, production of endothelin-1 (ET-1) and serum levels of some stress hormones such as possible pathogenetic factors of Raynaud's phenomenon and of scleroderma. This work ends with a proposal of a pathogenetic scheme that, even if it doesn't exclude an infective pathogenesis, considers stress as a factor able to switch on a series of psycho-neuro-immunoendocrinological reactions that can support, through the activation of lymphocytes, fibroblastic proliferation and finally fibrosis.

Long-term thymopentin treatment in systemic scleroderma.

A study was carried out to investigate whether thymopentin treatment is capable of inducing changes in the immunological status of patients with systemic scleroderma and to compare any such changes with modifications in clinical condition. Nine patients were given thymopentin, 1 ml in 10 ml saline solution, by slow intravenous infusion 3-times weekly for 5 weeks (cycle). The cycles were repeated at 3-month intervals. Treatment duration ranged from 1 to 5 years. Blood samples were drawn at the beginning and at the end of each cycle and the patients' lymphocytic sub-populations were examined. A control group of 9 comparable healthy subjects were similarly tested. Data analysis showed that a statistically significant decrease of CD16+ and CD25+ lymphocytes compared to pretreatment was already apparent at the end of the first thymopentin treatment cycle. An improvement was found in the clinical condition of 7 of the 9 patients at the end of the follow-up period with a significant correction of unbalanced lymphocytic subsets.

Rheological leukocyte indexes in diabetes mellitus type II.

Eleven healthy volunteers were examined, as well a thirty-five diabetic subjects divided into three groups according to the presence or absence of macro-microvascular complications and according to whether or not these complications were chronic. The results of our study show a marked unfavorable in cell rheology in the diabetic subjects, as studied using the St. George filtrometer. In particular, polymorphonuclear (PMN) cells, in the diabetic subject showed a substantial decreases in viscosity and elasticity, reflected by decreased IrFr and increased CR values. These changes were most marked in the group of diabetics with acute vascular events, i.e., AMI and stroke. Rheological indexes studied appeared to be independent from the simultaneous blood glucose level.

Early manifestations of dementing illness: treatment with glycosaminoglycan polysulfate.

1. In a multicenter, placebo-controlled, double-blind clinical trial in 156 elderly patients with psychopathologic symptoms, glycosaminoglycan polysulfate was found to be a therapeutically effective agent in the treatment of the earliest manifestations of a dementing process. 2. Treatment with glycosaminoglycan polysulfate in the daily dosage of 600 LRU, administered on the basis of a divided dosage schedule for 24 weeks, was significantly superior to an inactive placebo on several outcome measures including the SCAG Total and factor scores (i.e., Cognitive Dysfunction, Withdrawal, Agitation/Irritability and Depression), the NOWLIS Total and Fatiguability factor scores, the MMSE, the HAM-D Total and Vegetative Symptoms factor score and the CGI Severity of Illness and Global Improvement. 3. The drug was well tolerated; vital signs and laboratory measures did not show clinically significant changes within the experimental period.

Soluble interleukin-2 receptor, interleukin-2 and interleukin-4 in sera and supernatants from patients with progressive systemic sclerosis.

We studied the sera of patients with progressive systemic sclerosis (PSS) for elevated levels of soluble interleukin-2 receptor (sIL-2R), interleukin-2 (IL-2) and interleukin-4 (IL-4). We also measured IL-2, IL-4 and B cell growth factor (BCGF) activity in supernatants of peripheral blood mononuclear cells from the same patients. The finding of elevated serum sIL-2R and IL-2, and the increased levels of IL-2, IL-4 and BCGF activity in culture supernatants indicates that T lymphocyte hyperactivity likely play a major role in PSS. The failure to detect under our experimental conditions a direct proliferative effect of recombinant IL-2 on enriched normal B cells might suggest that IL-4 is the cytokine mainly responsible of the BCGF activity recovered in PSS supernatants.

Soluble CD8 antigen in systemic sclerosis.

In 15 patients suffering from systemic sclerosis (PSS) the function of CD8+ circulating lymphocytes was assayed by determining soluble CD8 antigen (sCD8) both in sera and in 48 hr PHA-conditioned media of peripheral blood mononuclear cells (PBMCs). Furthermore, the frequency of circulating activated CD8+ cells, which express DR antigens, interleukin-2 receptor, and transferrin-receptor, was determined by cytofluorographic analysis. The results of this preliminary study indicate that only in 6 PSS sera sCD8 was elevated as compared to healthy controls; furthermore, we found slightly increased sCD8 in culture media from PSS patients. The frequency of circulating activated CD8+ cells was similar both in PSS patients and in controls. Overall, our findings suggest that CD8+ cell activation is not a major phenomenon in PSS.

Polyclonal B lymphocyte activation in progressive systemic sclerosis.

In 13 women affected by progressive systemic sclerosis (PSS) the activation state of circulating B lymphocytes by use of monoclonal antibodies and the in vitro IgG, IgA, IgM synthesis by peripheral blood mononuclear cells (PBMC) cultured with or without pokeweed-mitogen (PWM) were evaluated. The total and "activated" B lymphocytes were increased. The supernatants of PBMC cultured without PWM contained increased IgG and IgA levels; only in two patients, however, were IgM levels increased. The addition of PWM augmented IgG and IgA levels only in three cases and never IgM levels. Our results indicate that the polyclonal hyperactivity of B lymphocytes is a major immunologic feature in PSS patients. The lack of increase of in vitro immunoglobulin synthesis in the presence of PWM suggests either that B lymphocytes, already maximally activated in vivo, are unresponsive to further mitogen stimulation or that a more complex imbalance of B lymphocyte responsiveness to PWM may exist.

Effects of prolonged picotamide therapy on platelet activity in patients with peripheral arterial disease.

An open study was carried out in 14 patients with peripheral arterial disease to investigate the effects of prolonged therapy with picotamide on platelet activity. Patients received daily oral doses of 900 mg picotamide for 1 month, 600 mg per day during the second month and 300 mg per day from the third to the sixth month of the study. Measurements were made before and during therapy of blood coagulation parameters and factors influencing platelet function, i.e. plasma beta-thromboglobulin and serum thromboxane B2. The results showed that there were no significant variations in platelet count, prothrombin time, partially activated thromboplastin time, presence and amount of fibrinogen in blood, and antithrombin III. Examination for fibrinogen degradation products was constantly negative and unaltered during therapy. Although plasma beta-thromboglobulin values did not vary significantly, there was a significant and progressive reduction throughout treatment in serum levels of thromboxane B2.

Haemorrheological improvement by pentoxifylline in patients with peripheral arterial occlusive disease.

A group of 25 patients with Stage II peripheral arterial occlusive disease was treated with pentoxifylline ('Trental' 400) for 90 days. Significant changes were observed during treatment in the haemorrheological parameters studied. At the end of treatment, reductions were found compared to basal values in blood viscosity (-10.1%), plasma viscosity (-9.5%), blood fibrinogen (-18.5%), and erythrocyte aggregation (-31.8%); blood filtrability was significantly increased (+41.0%). These positive changes in blood fluidity may be suggestive of beneficial influence of this drug on the clinical condition of claudicants.

[Clinical cases of primary hyperviscosity syndrome]. / Studio di alcuni casi clinici di sindrome da iperviscosità primaria.

Three cases of primary hyperviscosity syndrome with different etiology are described. For each of them the specific hemorheological characteristics which produce the common expression of blood hyperviscosity are emphasized.

[Statistical study on the effect of erythrocyte aggregation on blood filterability]. / Studio statistico sull'influenza dell'aggregazione eritrocitaria sulla filtrabilità ematica.

The values of erythrocyte aggregation (expressed as erythrocyte sedimentation rate corrected for plasma viscosity and hematocrit) were correlated with the values of blood filterability in a group of 165 healthy subjects and patients suffering for the most part from vascular disorders. A highly significant inverse correlation (r = -0.57, p less than 0.001) was observed between the two parameters, being a lower blood filterability correlated with a higher erythrocyte aggregation.

[Hemorrheological factors in the developmental stages of peripheral arterial occlusive diseases]. / Studio dei fattori emoreologici negli stadi evolutivi dell'arteriopatia ostruttiva periferica.

It was our aim to evaluate the degree of impairment of the blood rheological properties according to the clinical stage of peripheral occlusive arterial disease. From the 2nd to the 4th clinical stage the following hemorheological changes were observed: a progressive increase of whole blood viscosity, plasma viscosity, erythrocyte aggregation and plasma fibrinogen levels, as well as a progressive decrease of erythrocyte filterability.

[Multi-distributed ischemia and degree of hemorrheological changes]. / Ischemia pluridistrettuale e grado di alterazioni emoreologiche.

A hemorheological study has been carried out in patients suffering from peripheral occlusive arterial disease alone or associated with occlusive arterial disease of the epiaortic vessels or with ischemic heart disease. Significantly more severe hemorheological impairments of whole blood viscosity, plasma viscosity and erythrocyte filterability have been observed in patients with ischemia of two vascular districts than in patients with peripheral ischemia alone.

[Influence of age factors on the rheological properties of blood in healthy subjects and those with vascular disease]. / Influenza del fattore età sulle proprietà reologiche del sangue in soggetti sani e vasculopatici.

The main hemorheological factors (whole blood and plasma viscosity, hematocrit, erythrocyte aggregation and filterability, plasma fibrinogen concentration) have been examined in apparently healthy subjects and in vascular patients divided into different groups of age. Whereas in healthy subjects a progressive alteration of the hemorheological factors has been observed from adult to senile age, in vascular patients a similar behavior resulted only from the comparison between subjects of the 7th and the 8th decade of age and subjects of the 6th decade, while no significant changes were found between patients of the 7th decade and patients of the 8th decade of age.

[Hemorrheology and neurovascular syndromes of the extremities]. / Emoreologia e sindromi acrali neurovascolari.

The hemorheological factors of subjects affected by Raynaud's syndrome, by sclerodermia and by acrocyanosis have been compared with those of normal control subjects. The patients with Raynaud's syndrome and those with acrocyanosis, besides the phenomenon of critical ischemia, do not show any significant hemorheological difference in comparison with normal control subjects. In patients suffering from Raynaud's syndrome secondary to sclerodermic disease, an increased blood viscosity due to plasma hyperviscosity secondary to increased plasma fibrinogen levels, an increased erythrocyte aggregation and a reduced red blood cell filterability have been observed.

Haemodynamic and haemorheological effects of buflomedil in patients with peripheral occlusive arterial disease.

The haemodynamic and haemorheological effects of buflomedil after acute intravenous infusion and after chronic treatment were studied in 25 patients with occlusive arterial diseases of the lower limbs. Acute treatment (100 mg) induced an increase in ankle blood pressure and in peripheral blood velocity, mainly of the continuous flow, as well as a significant decrease in blood viscosity and a significant increase (p less than 0.01) in red cell deformability, expressed as erythrocyte filtration flow. Chronic treatment (200 mg/day for 20 days) showed haemodynamic and haemorheological effects similar to those of the acute infusion, with a highly significant increase (p less than 0.001) in the erythrocyte flow at the end of the treatment period.

[The immunity of senescence. Study of lymphocytes in vitro. I: Rosette forming lymphocytes]. / Immunità nella senscenza. Studio delle funzioni linfocitarie in vitro. I: Rosette linfocitarie.

E and EAC rosettes forming lymphocytes were evaluated in 25 elderly subjects (mean age 84.76 years) without manifestations of neoplastic or infectious diseases. The percentage of E rosettes appeared within the normal range in about 50 per cent of cases and below normal in remaining 50 per cent. No evident sex-related differences were observed. The percentage of EAC rosettes showed a larger distribution of values. In fact, 56 per cent of cases fell within normal limits, 26 per cent was higher and 20 per cent lower than normal. Our results are in agreement with the data reported in the most recent literature, which indicates that the main target of immune system senescence is the T-cell population, while B cells do not show significant age-related variations. The significant number of cases with percentages of EAC rosettes higher than normal may be attributed to the failure of immune homeostatic mechanisms normally exerted by T cell subpopulations. Further studies on several immune system functions in advanced age are in progress.