Franco Ameglio

CYTOKINES IN PSORIASIS

Many different stimuli can trigger the onset of psoriasis in genetically predisposed individuals.1 Although the disease was classified by Hebra as a distinct nosologic entity in 1841, the cell type(s) responsible for beginning the cascade of events leading to disease expression and the precise nature of the cell alteration are still unclear. Even with the new experimental approaches,2,3 no demonstration has yet been made of the possibility that the immunocyte or keratinocyte may be the principal actor in this disease and that the subsequent modifications may be mediated by different cytokines (see list of abbreviations in Table 1 and Fig. 1). Stimulated keratinocytes may act as initiators of an inflammatory process4 by means of the secretion of different modulators able to induce the expression of adhesion molecules on endothelial cells and the recruitment of circulating immunocytes (Fig. 1).4,5 These phenomena lead to the interaction between keratinocytes and activated T lymphocytes resulting in an increased proliferation of genetically predisposed keratinocytes.1,5 After stimulation, keratinocytes secrete a great variety of pro-inflammatory mediators, such as C5a, prostaglandins, leukotrienes, etc., also including many of the different cytokine types whose levels have been reported to be altered mainly at the level of psoriatic lesional skin (Table 2). These seem to play a key role in initiating and maintaining the two typical features of psoriatic lesions, i.e. keratinocyte hyperproliferation and inflammation (Table 3 and Fig. 1). Of the pro-inflammatory cytokines that are also produced by keratinocytes, the behavior of interleukin-1α (IL-1α), IL-1β, and tumor necrosis factor-alpha (TNF-α) in psoriasis will first be described. These three modulators, the socalled ‘‘primary cytokines,’’ activate a sufficient number of effector mechanisms to trigger independently cutaneous inflammation,6 and can be involved at an early stage in orchestrating the pathologic changes that occur in psoriasis.5

Granulocyte colony‐stimulating factor promotes the generation of regulatory DC through induction of IL‐10 and IFN‐α

We have recently demonstrated that G‐CSF promotes the generation of human T regulatory (TREG) type 1 cells. In this study, we investigated whether the immunomodulatory effects of G‐CSF might be mediated by DC. CD14+ monocytes were cultured with serum collected after clinical administration of G‐CSF (post‐G), which contained high amounts of IL‐10 and IFN‐α. Similar to incompletely matured DC, monocytes nurtured with post‐G serum acquired a DC‐like morphology, expressed high levels of costimulatory molecules and HLA‐DR, and exhibited diminished IL‐12p70 release and poor allostimulatory capacity. Importantly, post‐G DC‐like cells were insensitive to maturation stimuli. As shown by neutralization studies, IFN‐α and, even more pronounced, IL‐10 contained in post‐G serum inhibited IL‐12p70 release by post‐G DC‐like cells. Furthermore, phenotypic and functional features of post‐G DC‐like cells were replicated by culturing post‐G monocytes with exogenous IL‐10 and IFN‐α. Post‐G DC‐like cells promoted Ag‐specific hyporesponsiveness in naive allogeneic CD4+ T cells and orchestrated a TREG response that was dependent on secreted TGFβ1 and IL‐10. Finally, neutralization of IL‐10 and IFN‐α contained in post‐G serum translated into abrogation of the regulatory features of post‐G DC‐like cells. This novel mechanism of immune regulation effected by G‐CSF might be therapeutically exploited for tolerance induction in autoimmune disorders.

Association between cutaneous melanoma, Breslow thickness and vitamin D receptor BsmI polymorphism

Summary  Background  Literature data report an association between some vitamin D receptor (VDR) polymorphisms and different kinds of tumours, including malignant melanoma (MM). Only three VDR polymorphisms (FokI, TaqI and A‐1012G) have been investigated in association with the presence of cutaneous MM or the development of metastases.

Endothelin-1 Levels Are Increased in Sera and Lesional Skin Extracts of Psoriatic Patients and Correlate with Disease Severity

Endothelins (ETs), in addition to their systematical activities, exert important functions at the skin level, such as increase of keratinocyte proliferation, neo-angiogenesis and leukocyte chemotaxis, which are among the main characteristics of psoriasis. To assess a possible ET-1 involvement in plaque-type psoriasis, ET-1 determinations were carried out in 15 sera and 8 lesional and non-lesional biopsy skin extracts from psoriatic patients and in 15 sera and 5 biopsy skin extracts from healthy volunteers, sex- and age-matched, using commercially available ELISA kits. A statistical analysis of the results showed that ET-1 levels were increased in sera of psoriatic patients, as compared to normal subjects (p = 0.04). In addition, there was a significant correlation between both serum (r = 0.60, p = 0.02) and lesional skin (r = 0.80, p = 0.03) ET-1 values versus the Psoriasis Area and Severity Index scores. Significant increases of the lesional versus the non-lesional (p = 0.01) and versus the normal (p = 0.04) ET-1 skin extract values were observed, together with a significant correlation between lesional and non-lesional ET-1 skin levels (r = 0.79, p = 0.03). These findings were also confirmed at the mRNA level, using RT-PCR analysis, where increased ET-1 mRNA levels, densitometrically measured, were found in the lesional samples versus non-lesional and normal skin. Since interleukin-8 is involved in psoriasis and shares some biological properties with ET-1, we further evaluated the levels of this cytokine in skin extracts. The behaviour of interleukin-8 paralleled that of ET-1, and a significant correlation between these two molecules was observed in the lesional skin (r = 0.76, p = 0.05). Taken together, these data stress that, as previously described for interleukin-8, ET-1 may be involved in inflammatory processes associated with psoriasis.

Very high-dose cisplatin-induced ototoxicity: a preliminary report on early and long-term effects

SummaryAcute and subacute audiometric hearing changes were evaluated in 12 patients receiving 35 courses of very high-dose (vhd) cisplatin (200 mg/m2 per course) in hypertonic saline at 4 or 8-week intervals. Audiogical evaluations were performed both before and immediately after each course of chemotherapy, and again after the discontinuation of treatment. A significant drop of the mean hearing threshold (P<0.01) at high frequencies was observed even within 48 h from the end of the first course of therapy, with 50% of the patients presenting a hearing loss of more than 15 dB. At the same total dose (200 mg/m2), one course of this regimen provided an incidence of hearing loss of more than 15 dB, which was four times greater than that reported with two courses of standard-dose regimens. The incidence and severity of the hearing impairment progressed further with subsequent courses of chemotherapy. Compared with baseline levels, most patients (75%) receiving at least two courses had a moderate to severe hearing loss, especially involving 4 and 8 kHz. At the end of treatment, 33% of the patients complained of a nondisabling functional hearing impairment. No recovery occurred after chemotherapy had been discontinued for 9–28 weeks. At this dose level cisplatin is markedly ototoxic. The use of hypertonic saline and vigorous hydration are effective means of minimizing the risk of nephrotoxicity, but seem to have no effect on cisplatin-related ototoxicity.

Discriminant analysis on small cell lung cancer and non-small cell lung cancer by means of NSE and CYFRA-21.1

A correct diagnosis of small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) is essential both for prognostic and therapeutic reasons. We used discriminant analysis as a method to optimize the discriminant power of serum tumour marker levels for differentiation between SCLC and NSCLC. A panel of serum markers, including neurone specific enolase (NSE), cytokeratin fragment antigen 21.1 (CYFRA-21.1), tissue polypeptide antigen (TPA) and carcinoembryonic antigen (CEA) was obtained in 50 consecutive NSCLC and 17 SCLC. Data were analysed by the BMDP statistical program after logarithmic transformation of marker levels. The variables selected were NSE and CYFRA-21.1. Considered together, they were able to give a 97% rate of correct classification. The formula generated (canonic variable, CV) was validated on a group of seven SCLC and 22 NSCLC patients. Only two errors occurred. We therefore conclude that the canonic variable tested, based on NSE and CYFRA-21.1, provides a good discrimination between the two types of lung cancer. The method is rapid, relatively inexpensive, and based on simple serum tests.

Increased serum IL‐6, TNF‐alpha and IL‐10 levels in patients with bullous pemphigoid: relationships with disease activity

AIM The present report analyzes the serum levels of three cytokines, interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) and interleukin-10 (IL-10) in 15 patients with bullous pemphigoid (BP) (compared with 20 healthy controls) to evaluate a possible involvement of these biological modulators in the clinical expression of this disease. BACKGROUND BP is a rare bullous disease of autoimmune origin with evidence of inflammatory processes that cause skin lesions with local increase of various pro-inflammatory mediators. METHODS Determination of cytokine concentrations were obtained employing commercially available ELISA kits. RESULTS The sera of BP patients showed increased levels of these three cytokines (P < 0.01). When the number of skin lesions (blisters and/or erosion) of each patient, employed as a marker of disease activity, was correlated with the serum levels of IL-6 and TNF-alpha, significant correlations were found (IL-6: P < 0.01 and TNF-alpha: P < 0.01, respectively), suggesting a possible role of these mediators in the development of BP blisters. The serum levels of IL-6 also correlated (P = 0.01 with those of serum C reactive protein (CRP), an acute-phase protein induced by IL-6 in hepatocytes. In addition, serum TNF-alpha and sE-selectin (an adhesion molecule previously reported to be increased by this cytokine) levels were also correlated (P < 0.05). CONCLUSIONS On the basis of these data, it may be indicated that at least IL-6 and TNF-alpha are associated with the clinical expression of BP and that the endothelial activation (possibly induced by the TNF-alpha activity), seems to be an important phase of this dermatosis.

Recognition and treatment of psoriasis. Special considerations in elderly patients.

SummaryPsoriasis is a chronic dermatological disorder that affects 1 to 2% of the general population. Although its aetiology is still unknown, the importance of genetic factors has been confirmed by many studies, mainly in young individuals. With respect to clinical features, plaque-type psoriasis (localised or generalised) is the most common form. At present, there is no cure for psoriasis and the available treatments can only temporarily clear the skin manifestations.The choice of treatment regimen for psoriasis is based on the severity of the disease, the patient’s gender, age, treatment history and level of compliance, and the physician’s personal experience. All therapies for psoriasis have different and potentially toxic effects. Therefore, a good knowledge of their relative and absolute contraindications, adverse effects and interactions with other drugs is mandatory.The elderly represent a significant proportion of patients with psoriasis because its prevalence increases with age. Physicians, particularly general practitioners, dermatologists and gerontologists, must be aware of the problems that the treatment of psoriasis in the elderly can present. This is especially important because of the increased risk of adverse drug reactions in the elderly.

GSTM1-null polymorphism as possible risk marker for hypertension: results from the aging and longevity study in the Sirente Geographic Area (ilSIRENTE study)

BACKGROUND Involvement of various gene mutations in hypertension have been already reported, including GST (detoxification Phase II enzymes), with contrasting results. This study analyzes a possible association between GSTM1-null and GSTT1-null polymorphisms and the hypertension status in a very old population (>80 years). METHODS 354 old patients were collected (255 were hypertensive and 99 were not). RESULTS GSTM1-null individuals (n=156) were significantly associated with increased risk of hypertension [OR=2.25 (1.36-3.72); p=0.005]. This association was confirmed both in 115 male and 239 female subjects. In contrast, no significant association was observed [p=0.52, OR=1.24 (0.67-2.29)] in the 57 hypertensive GSTT1-null genotypes vs 198 wild-type individuals. The above mentioned significances were obtained by multivariate logistic regression analysis after adjusting for confounder variables. Alcohol consumption and/or smoke habits were not significantly different between the two groups, as well as gender, marital status, education, number of concomitant diseases, and presence of cardiovascular diseases. No synergistic association was observed for the combined null genotypes of GSTT1 and GSTM1. CONCLUSIONS The knowledge of GSTM1 variant status seems to be potentially useful to predict a possible hypertensive status after 80 years of age. This study underlines a possible importance of the GSTM1 enzyme for blood pressure regulation.

Identification of a new control region in the genome of the DDP strain of BK virus isolated from PBMC

The various strains of human polyomavirus BK (BKV) show a marked heterogeneity in the noncoding control region (NCCR), which includes the origin of replication and the regulatory region for early and late transcription. A new BKV strain (DDP, U91605) was identified by direct detection and sequencing of PCR products of BKV‐NCCR DNA obtained from PBMC samples of HIV‐positive or ‐negative subjects. The DDP strain NCCR sequence showed an organisation not described previously in vivo with the maximum homology with the archetypal strain (WW) (M34048), as compared with those collected in GenBank. Structurally, P68, Q39, and S68 boxes were perfectly conserved, whereas the R63 box was completely deleted. This deletion involves the loss of sequences able to bind cellular factors essential for the DNA transcription, such as NF1 binding sites, normally present twice in the R box and the modification of SP1. It is possible that these rearrangements represent a cause of the loss of the VP1 region observed in 9/22 PBMC samples and never observed in urine isolates, which are similar to the WW strain. J. Med. Virol. 58:413–419, 1999.