Renato Carignola

Inhibition by cortisol of human natural killer (NK) cell activity.

The effects of cortisol on the natural killer (NK) activity of human peripheral blood mononuclear (PBM) cells were studied in vitro using a direct 4-h 51Cr-release assay and K 562 cell line as a target. Preincubation for 20 h of PBM cells drawn from healthy donors with 1 X 10(-8) to 1 X 10(-5) M cortisol resulted in a significant decrease of NK cell activity. The magnitude of the suppression was directly related to the steroid concentration and inversely related to the number of effector cells. Cortisol was able to minimize the enhancement of NK cytotoxicity obtainable in the presence of immune interferon (IFN-gamma). A significantly higher suppression was achieved after sequential exposure of PBM cells to cortisol and equimolar levels of prostaglandin E2 (PgE2). The concomitant incubation with theophylline and isobutyl-methylxanthine failed to enhance the cortisol-induced suppression, whereas PgE2-dependent inhibition significantly increased after exposure of PBM cells to methyl-xanthines. The inhibitory effect of cortisol was partially or totally prevented by the concomitant incubation with equimolar amounts of 11-deoxycortisol and RU 486 but not of progesterone. Treatment of NK effectors with a monoclonal anti-human corticosteroid-binding globulin (CBG) antibody produced an enhancement of the spontaneous NK activity and a partial suppression of cortisol-mediated effects. Our results suggest that endogenous glucocorticoids play a role in the regulation of NK cell-mediated cytotoxicity. Since the effect of cortisol was additive to that of PgE2 and was not changed by phosphodiesterase inhibitors, it is conceivable that the hormone acts at a level different from the adenylate cyclase-phosphodiesterase system. Data obtained with the use of antiglucocorticoids and the anti-CBG antibody are compatible with a role both of high-affinity glucocorticoid receptors and of CBG in mediating cortisol action on the human NK cell activity.

Hypovitaminosis D in Systemic Sclerosis

To the Editor: We read with interest the articles by Dr. Braun-Moscovici, et al 1 and Vacca, et al 2 reporting a strikingly high prevalence of 25-hydroxyvitamin D (25OHD) deficiency in patients with systemic sclerosis (SSc). Their remarkable findings confirm the results of our work, to our knowledge the first controlled study to demonstrate an increased prevalence of hypovitaminosis D in SSc3. The renewed interest in vitamin D in autoimmune diseases gives us the opportunity to show previously unpublished data and make some observations about this issue. Our study comprised 60 patients and 60 matched controls3. 25OHD levels were lower in patients than in controls [median 23 ng/ml (range 3–92) and 39 ng/ml (14–138), respectively; p < 0.001] even after matching for season of sampling. Thirty-eight patients had 25OHD levels below 30 ng/ml; 4 patients fell in the severe deficit range (< 10 ng/ml), 17 in the insufficiency range (≥ 10 and < 20 ng/ml), and 17 in the mild hypovitaminosis range (≥ 20 and < … Address correspondence to Dr. Calzolari; E-mail: gilberto983{at}gmail.com

Circulating Osteoprotegerin and Soluble RANK Ligand in Systemic Sclerosis

Objective Microvascular damage is an early pathogenetic event in systemic sclerosis (SSc). The receptor activator of nuclear factor-κB ligand (RANKL)/RANK/osteoprotegerin (OPG) system is involved in vascular biology. Our aim was to assess OPG and soluble RANKL (sRANKL) serum levels in patients with SSc and healthy controls. Methods Sixty patients with SSc (median age 58, range 31–72 yrs) and 60 healthy subjects matched for age, sex, and menopausal status were recruited. Serum OPG, sRANKL, soluble vascular cell adhesion molecule (sVCAM; marker of endothelial activation/injury), and bone turnover markers were measured. Bone mineral density in patients was assessed and cardiovascular/coronary risk was estimated. Results OPG was similar in the 2 groups, while sRANKL and sRANKL/OPG ratio was higher in patients (p = 0.01 for both). sVCAM was markedly higher in patients (p < 0.001). OPG levels correlated positively with age in both patients (Spearman R = 0.50, p < 0.001) and controls (R = 0.56, p < 0.001). In patients, OPG was lower in men and higher in those with active ulcers or calcinosis. sRANKL levels were higher in patients treated with platelet aggregation inhibitors, and correlated negatively with densitometric measures. 25-hydroxyvitamin D levels were significantly lower in patients (p < 0.001). In patients, OPG levels correlated positively with cardiovascular and coronary risk (R = 0.28, p = 0.05 and R = 0.34, p < 0.01, respectively) and were higher in patients with hypertension and left ventricular hypertrophy. sVCAMlevels correlated positively with cardiovascular and coronary risk (R = 0.27, p = 0.06, and R = 0.38, p < 0.01, respectively). Conclusion Higher sRANKL levels and sRANKL/OPG ratio in patients with SSc are likely to be a consequence of altered bone microenvironment. We show a dissociation between the well established marker of endothelial activation/injury, sVCAM, and the alleged marker of vascular damage, OPG, in patients with SSc. Further studies are needed to better ascertain the relationships of the RANKL/RANK/OPG system with the progression of macro- and microvascular damage.

Studies on the mechanism of cortisol inhibition of human natural killer cell activity: effects of calcium entry blockers and calmodulin antagonists

The role of Ca2+ in mediating the inhibition by glucocorticoids of human natural killer (NK) activity was investigated using Ca2+ entry blockers (verapamil and its desmethoxy-derivatives LU46973 and LU47093) and calmodulin antagonists (pimozide and two naphthalenesulfopamide derivatives, W-7 and W-13). Peripheral blood mononuclear (PBM) cell preparations were incubated for 20 h with 1 x 10(-6) M cortisol and these agents in various combinations (concentration range: 1 x 10(-7) - 1 x 10(-5) M) and then assayed in a direct 4-h cytolytic assay using 51Cr-labeled K 562 target cells. Exposure to cortisol led to a significant reduction of NK cell activity (about 50% with respect to the spontaneous activity). Ca2+ entry blockers displayed per se a dose-dependent depressive effect on cytotoxicity and gave significant enhancement of cortisol-dependent inhibition. Calmodulin antagonists were per se minimally effective but clearly amplified the cortisol-mediated inhibition. Raising extracellular Ca2+ by CaCl2 or intracellular Ca2+ by the ionophore A23187 yelded an appreciable reduction of these effects. Our data are compatible with the view that extracellular and intracellular Ca2+ play a role in the control of human NK cell activity. Moreover, it is conceivable that the mechanisms involved in glucocorticoid inhibition of NK cell activity involve Ca2+-dependent pathways.

Seasonal effects of rotational stress on Lewis lung carcinoma metastasis and T-lymphocyte subsets in mice.

Rotational stress specifically increases the formation of spontaneous lung metastasis in mice bearing Lewis lung carcinoma, without significantly modifying the growth of primary tumor. The increase in metastasis number and volume caused by rotational stress varies in magnitude with a highly significant circannual rhythm; the acrophase approximately coincides with summer solstice. Rotational stress causes a significant reduction in the number of CD3+ and CD4+ T-lymphocyte subsets in summer, whereas in winter the number of CD3+ subset is significantly increased; the CD4+/CD8+ ratio and the number of NK 1.1 antigen positive cells are not significantly modified by rotational stress in both periods considered. The increase in metastasis formation by rotational stress thus appears to negatively correlate with the number of splenic CD3+ and CD4+ T-lymphocyte subsets. This seasonal behavior occurs in spite of the control of light cycle, temperature and humidity in the animal housing, suggesting the existence in the host of an endogenous oscillator with a circannual period. These data indicate the opportunity to consider endogenous rhythms within the host, as well as seasonal factors, in studies on stress and neuroimmunomodulation in experimental oncology.

Vitamin D Status and Quality of Life in Systemic Sclerosis Patients.

To the Editor: R ecently, we read with great interest the article by Raczkiewicz et al, reporting that low serum vitamin D levels (25OHD) are associated with active disease as well as to a worse quality of life in rheumatoid arthritis patients. Hypovitaminosis D has been previously described also in systemic sclerosis (SSc), and 25OHD deficiency was associated with disease severity measured by skin involvement, cutaneous calcinosis, disease duration and activity, pulmonary artery pressure (PAP), and diffusing capacity of the lung for carbon monoxide (DLCO) (Table 1). In a previous study, we measured serum levels of 25OHD in 60 SSc patients: 25OHD was less than 20 ng/mL in 35% of them (21/60); a lack of any significant correlation with disease characteristics (limited or diffuse form, gastrointestinal involvement, cutaneous ulcers joint involvement) was observed. 25-Hydroxyvitamin D serum levels showed some correlation with

Initial combination therapy for patients with pulmonary arterial hypertension (PAH): a budget impact analysis from the perspective of the Italian national healthcare system

ABSTRACT Background: Initial combination therapy in patients with pulmonary arterial hypertension (PAH) WHO functional class (FC) II or III has demonstrated clinical benefits over initial monotherapy. The objective of this study is to compare the financial impact of initial combination therapy with initial monotherapy for incident patients with PAH in Italy. Methods: A 3-year budget impact model compared a ‘status quo’ scenario of initial monotherapy with an endothelin receptor antagonist (ERA), phosphodiesterase 5 inhibitor (PDE5i) or prostanoid, with a ‘new’ scenario involving initial combination therapy, using Italian national healthcare system (NHS) data for incident patients with PAH WHO FC II or III. The hospitalisation hazard ratio (HHR) from the AMBITION study and expert panel advice on therapy use were employed. Univariate sensitivity analyses were performed. Results: A difference in costs of €16,070 favouring the ‘new’ scenario (initial combination therapy) was observed, and attributed to 101 fewer hospitalisations over 3 years. Sensitivity analyses showed that costs were driven by the proportion of patients receiving ERAs versus PDE5i, hospitalisation costs and prostanoid dose. Conclusion: Initial combination therapy instead of monotherapy could reduce the number and cost of hospitalisations without an increase in the total costs to the Italian NHS.