Long-term clinical effects of interferon gamma-1b and colchicine in idiopathic pulmonary fibrosis.

Idiopathic pulmonary fibrosis (IPF)/usual interstitial pneumonia is a deadly disease with no effective treatment. The purpose of this randomised prospective multicentric study was to characterise the clinical effects of interferon gamma (IFN-gamma) 1b administered subcutaneously thrice weekly versus colchicine for 2 yrs. This study had no pre-specified end-points. Fifty consecutive IPF patients were randomised. Patients with mild-to-moderate IPF were eligible for the study if they had histologically proven IPF, or, in the absence of surgical biopsy, fulfilled the European Respiratory Society/American Thoracic Society criteria. In the intent-to-treat population, five out of 32 (15.6%) IFN-gamma-1b patients and seven out of 18 (38.8%) colchicine patients died after a median follow-up period of 25 months Patients treated with IFN-gamma 1b showed a better outcome after 2 yrs of therapy, and fewer symptoms, as assessed using the St George's Respiratory Questionnaire, after 12 months of therapy. Also, the IFN-gamma-1b group exhibited a higher forced vital capacity (percentage of the predicted value) after 24 months of treatment. No significant differences were detected in resting arterial oxygen tension, total lung capacity (% pred), transfer factor of the lung for carbon monoxide (% pred) and high-resolution computed tomographic scoring between the two treatment groups. These data suggest that long-term treatment with interferon gamma 1b may improve survival and outcome in patients with mild-to-moderate idiopathic pulmonary fibrosis. Further studies are needed to verify these results.

Sarcoidosis in untreated chronic hepatitis C virus infection.

Chronic hepatitis C has been implicated in the pathogenesis of sarcoidosis in several cases of patients treated with interferon-alpha. On the other hand, only in a few cases previously has a possible link between sarcoidosis and untreated chronic hepatitis C virus infection been demonstrated. We report on a patient with chronic hepatitis C who developed cutaneous sarcoidosis without prior interferon-alpha treatment. We hypothesize that viral persistence seen in chronic hepatitis C virus infection might be one of the potential factors that trigger cellular immune response in granulomatous reactions as seen in sarcoidosis, in genetically predisposed patients.

Assessment of mycobacterial, propionibacterial, and human herpesvirus 8 DNA in tissues of greek patients with sarcoidosis.

The causes of sarcoidosis are unknown. In this study, we report the presence of Mycobacterium tuberculosis complex and Propionibacterium granulosum DNA in a significant proportion of Greek patients with sarcoidosis. Human herpesvirus 8 DNA was not detected in sarcoid tissues from Greek patients. Our findings are discussed.

Aberrant p16 expression is correlated with hemizygous deletions at the 9p21-22 chromosome region in non-small cell lung carcinomas.

The p16 protein is encoded by the CDKN2 gene, and functions as an inhibitor of cyclin-dependent kinase 4 and 6 (CDK4/6). Phosphorylation of the retinoblastoma protein (pRb) by CDK4/6 represents a vital step in cell cycle progression. Alterations of p16INK4A are frequent events in human malignancies. In non-small cell lung carcinoma (NSCLC) the data concerning the mechanisms of p16INK4A inactivation suggest that point mutations and aberrant methylation of its promoter can only account for a proportion of the cases with abnormal p16 immunoexpression. The role of deletions in this procedure is not yet clarified. In order to gain more insight into the role of deletions in p16INK4A deregulated expression, we investigated the state of the chromosomal region 9p21-22 in a series of 57 NSCLCs, by performing a detailed mapping analysis, using a tight cluster of highly polymorphic microsatellite markers, and correlating the findings with p16 immunostaining. Abnormal p16 expression was observed in 46% of the NSCLCs examined. No relationship was observed between p16 abnormal staining and various clinicopathological parameters. Abnormal p16 protein staining was strongly associated with hemizygous deletions at the IFNA and D9S171 microsatellite loci, which demarcate the region encoding the p16INK4A gene (P = 0.002). These findings suggest that deregulated expression of p16 is involved in the multistage process of NSCL carcinogenesis and that deletions may represent a predominant mechanism of p16INK4A inactivation. A significant percentage also of LOH was noticed at the D9S162 (35%) and D9S126 (38%) loci which lie 6cM and 4cM, respectively, far from the area which encodes p16INK4A, implying that other tumor suppressor genes (TSGs) may reside in this region. Although the overall incidence of LOH at the examined region was high (58%), we did not observe any correlation with smoking habits, histology and lymph node status. Another noteworthy finding was the existence of microsatellite instability (MI) in 11% of the patients. MI provides a marker for replication error phenotype (RER+), a recently defined manifestation of genetic instability observed in a wide range of tumors. In conclusion, alterations (LOH + MI) at the 9p21-22 chromosome region are frequent events in NSCLCs and may affect directly or indirectly the expression of p16.

Immunohistochemical and molecular evaluation of the mdm-2 gene product in bronchogenic carcinoma.

In this study, we investigated immunohistochemically the expression of mdm-2 protein in 93 surgically resected bronchogenic carcinomas. The findings were correlated with p53 protein detection status and clinicopathologic data (histologic type, differentiation grade of the lesions, lymph node metastases, and smoking history of the patients). Thirty of the 93 immunohistochemically examined specimens were subjected to Northern blot and differential polymerase chain reaction analysis to look into the mechanism of mdm-2 overexpression. Finally, we studied the concordance between p53 immunohistochemical positivity and p53 gene alterations as assessed by the single-strand conformation polymorphism technique. Seventy-three (78%) and 67 (72%) of 93 carcinomas showed nuclear immunoreactivity for mdm-2 and p53 proteins, respectively. We observed a high degree of concordance (75%) between p53 mutations and p53 immunolabelling, which was even higher in the specimens with p53 positively in more than 50% of the cells (90%). Despite the high percentage of mdm-2 and p53 expression, the two molecules were simultaneously detected in 50 (54%) of 93 cases. Forty-two (84%) of the 50 cases were accompanied by p53 mobility shifts, which indicated mutations. Interestingly, statistical analysis revealed an almost significant correlation between the carcinomas with mdm-2/p53 coexpression and lymph node disease (P = 0.058), which indicated a possible "gain of function" phenotype. In addition, absence of reactivity for both proteins was statistically more frequent in the patients without lymph node disease (P = 0.006). The mdm-2-positive/p53-negative immunohistochemical profile was more often seen in adenocarcinomas (P = 0.003), especially in well-differentiated ones (P = 0.02), than in other histologic types of lung cancer, which suggested a p53-independent pathway of mdm-2 overexpression. Molecular analysis showed that mdm-2 overexpression was a consequence of increased transcription rather than of mdm-2 gene amplification. The smoking history of the patients was strongly related to p53 (P = 10(-4)) even in the group of adenocarcinomas (P = 0.012). No correlation was observed between cigarette consumption and mdm-2 immunoreactivity.

Molecular and immunohistochemical study of class I growth factor receptors in squamous cell lung carcinomas.

The class I growth factor receptor family includes epidermal growth factor receptor, i.e. c-erbB-1, c-erbB-2 and c-erbB-3 molecules. These receptors have a significant sequence homology and play an important role in cell growth and differentiation. To further investigate their implication in squamous cell lung carcinomas (SqCLCs), we studied the protein expression by immunohistochemistry and examined for possible gene amplification by a novel semi-quantitative differential polymerase chain reaction (DPCR) technique. Expression of c-erbB-1, c-erbB-2 and c-erbB-3 was present in 65%, 28% and 10% respectively, of 40 SqCLCs cases. Seven of the 11 cases that expressed c-erbB-2, as well as all 4 c-erbB-3 expressing cases, also stained with the anti-c-erbB-1 mAb. Expression of c-erbB-1, but not of c-erbB-2 or c-erbB-3, correlated with the grade of tumor differentiation (100%, 64% and 36% positive cases of well, moderately and poorly differentiated cases respectively, p < 0.003). In addition, c-erbB-1 expression correlated with the presence of regional lymph node metastases within the moderately differentiated group. The c-erbB-1 gene was amplified in 11/40 (28%) cases, all of which overexpressed c-erbB-1 protein, while c-erbB-2 gene amplification was detected in only one case. There was no c-erbB-3 gene amplification in any of the 40 SqCLCs cases. These findings suggest that c-erbB-1, c-erbB-2 and c-erbB-3 receptors do not have a common role and are of different physiological importance, at least at the stage of clinically overt tumor in human SqCLCs.

Epidermal growth factor receptor expression in squamous cell lung carcinomas: an immunohistochemical and gene analysis in formalin-fixed, paraffin-embedded material.

Epidermal growth factor (EGF) and its receptor (EGFr) constitute an important and well-characterized mitogenic system in various ectodermal tissues. We evaluated the expression of EGFr and examined possible EGFr gene alterations in 18 formalin-fixed, paraffin-embedded squamous cell lung carcinomas (SCLC) by an immunohistochemical assay, Southern blotting and differential polymerase chain reaction (DPCR). The immunohistochemical study employing the F4 and EGF-R1 monoclonal antibodies, directed against the intra- and extra-cellular portion of the receptor respectively, showed EGFr over-expression in 89% of the SCLC cases examined. All cases showed positive immunostaining for both antibodies, thus excluding the possibility of truncated receptors. In addition, analysis of the EGFr gene was carried out by Southern blotting and DPCR on paraffin extracted DNA from the same carcinoma cases. We found amplification of the EGFr gene in 5/18 (27%) SCLCs. All 5 positive cases showed EGFr over-expression, suggesting a possible correlation between the presence of EGFr gene amplification and over-expression of receptor protein. No correlation was observed among EGFr staining, EGFr gene amplification and differentiation of carcinomas. In addition, Southern blot analysis with HER-A2, a probe which hybridizes a sequence of the receptor's intracellular domain, revealed three novel EcoRI restriction fragment patterns. We suggest that these patterns correspond to EcoRI polymorphic sites of the receptor's tyrosine kinase domain.