ICAM-1 expression by lung cancer cell lines: effects of upregulation by cytokines on the interaction with LAK cells.

Intercellular adhesion molecule-1 (ICAM-1) expression by tumour cells may be involved in their interaction with defensive cells. In this study the surface ICAM-1 expression and soluble ICAM-1 (sICAM-1) production by five small cell lung cancer (SCLC) and five non-SCLC (NSCLC) cell lines was investigated. In addition, the effects of ICAM-1 upregulation by cytokines on the adhesion of lung cancer cells to allogeneic lymphokine-activated killer (LAK) cells and susceptibility to LAK cytotoxicity was also evaluated. ICAM-1 expression was assessed by flow cytometry. Soluble ICAM-1 release was measured by enzyme-linked immunosorbent assay (ELISA). Interaction with LAK cells was tested by adhesion and cytotoxicity assays. At baseline, SCLC lines did not express ICAM-1, while 4 of the 5 NSCLC lines expressed ICAM-1. ICAM-1 expression was induced by interferon-gamma (IFN-gamma) in 4 of the 5 SCLC lines and upregulated in 1 of the 5 NSCLC lines. ICAM-1 expression was induced by tumour necrosis factor-alpha (TNF-alpha) in 1 of the 5 SCLC lines (National Cancer Institute (NCI) H211), and upregulated in 2 of the 5 NSCLC lines (NCI H460 and NCI H838). Among the latter lines, one (NCI H838) released significant amounts of sICAM-1. Adhesion to LAK cells and susceptibility to LAK cytotoxicity were significantly higher in TNF-alpha-treated NCI H460 and NCI H211 cells, compared to untreated NCI H460 and NCI H211 cells. In contrast, no difference in adhesion to LAK cells and susceptibility to LAK cytotoxicity was detected between baseline and TNF-alpha-treated NCI H838 cells. Intercellular adhesion molecule-1 surface expression and soluble intercellular adhesion molecule-1 release may play an important role in interactions between lymphokine-activated killer cells and lung cancer cells.

Morphologic and phenotypic changes of human neuroblastoma cells in culture induced by cytosine arabinoside.

The effects of cytosine-arabinoside (ARA-C) on the growth and phenotypic expression of a new human neuroblastoma (NB) cell line (GI-ME-N) have been extensively tested. Low doses of ARA-C allowing more than 90% cell viability induce morphological differentiation and growth inhibition. Differentiated cells were larger and flattened with elongated dendritic processes; such cells appeared within 48 h after a dose of ARA-C as low as 0.1 micrograms/ml (about 1000-fold lower than the conventional clinic dose). The new morphological aspect reached the maximum expression after 5-6 days of culture being independent from the addition of extra drug to the culture. A decrease in [3H]thymidine incorporation was also observed within 24 h and the cell growth was completely inhibited on the sixth day. Moreover, ARA-C strongly inhibited anchorage-independent growth in soft agar assay. Membrane immunofluorescence showed several dramatic changes in NB-specific antigen expression after 5 days of treatment with ARA-C. At the same time ARA-C also modulated cytoskeletal proteins and slightly increased catecholamine expression. These findings suggest that noncytotoxic doses of ARA-C do promote the differentiation of GI-ME-N neuroblastoma cells associated with reduced expression of the malignant phenotype.

Role of Broviac catheters in infections in children with cancer.

In a 3-year period 157 single lumen Broviac catheters were inserted in 145 children with various neoplastic diseases. The overall duration of the catheter courses was 30,533 days (median, 171; range, 2 to 647). Sixty-five percent of the catheter courses (102 of 157) were complicated by at least 1 febrile episode, for a total of 157 episodes. According to European Organization for Research on Treatment of Cancer criteria, 79 febrile episodes (50%) were classified as microbiologically documented infections, 57 (36%) with and 22 (14%) without septicemia; 31 (20%) as clinically documented infections; and 47 (30%) as possible infections. Of the 79 microbiologically documented infections 21 were catheter-related infections (CRI), 32 were catheter-unrelated infections and 26 were infections of unknown source. Only 48% of CRI occurred during neutropenia (less than 1000 neutrophils/mm3), compared with 88% of catheter-unrelated infections and 96% of infections of unknown origin (P = 0.00007). Gram-positive microorganisms (56% staphylococci) accounted for 78% of all isolates in CRI, 47% in catheter-unrelated infections and 43% in infections of unknown origins (P = 0.03). Fungi represented 12% of all isolates. Clinical and microbiologic resolution without removal of the catheter was achieved in 12 of 21 CRI (57%) and no patient died from a CRI. This study indicates that over 3 of 4 of CRI are caused by Gram-positive bacteria, occur in neutropenic and non-neutropenic patients (approximately 50%) and can be successfully treated without removing the catheter.

Cytogenetic and molecular study of two human neuroblastoma cell lines.

Long-term cell cultures (GI-LA-N and GI-ME-N) were established from the metastases of two disseminated neuroblastomas (NB). The first was obtained from a lymph node biopsy of a stage III NB after 7 months of chemotherapy, and the second from a bone marrow specimen of a stage IV NB after 6 months of chemotherapy. Cytogenetic investigation revealed several structural and numerical alterations in both cell cultures, but the only common chromosomal aberration was partial monosomy of 1p. Moreover, at the 5th in vitro passage, GI-LA-N displayed a high number of double minutes, not seen in GI-ME-N even after 33 subcultures. Molecular analysis revealed N-myc oncogene amplification in GI-LA-N cells, whereas, only one copy was found in GI-ME-N. No structural N-myc rearrangement was detected in either cell culture.

Effect of cytosine arabinoside on the growth and phenotypic expression of GI-ME-N, a new human neuroblastoma cell line.

Cytosine-arabinoside (ARA-C) effects on a new human neuroblastoma cell line (GI-ME-N), recently established in our laboratory, have been extensively tested. Low doses of ARA-C allowing virtually 100% cell viability induce morphological differentiation and growth inhibition; differentiated cells appear larger and flattened with elongated dendritic processes; such cells appeared within 48 hours after a dose of ARA-C as low as 0.1 microgram/ml. The new morphological aspect reached the maximum expression after 5-6 days of culture being independent of the addition of extra drug to the culture. A decrease in (3H)-thymidine incorporation was also observed within 48 hours being the cell growth completely inhibited at the 6th day. Membrane immunofluorescence with specific monoclonal antibodies showed several dramatic changes in NB-specific antigen expression after 5 days of treatment with ARA-C. These findings suggest that low ARA-C doses promote the differentiation of GI-ME-N neuroblastoma cells resulting in an altered expression of the malignant phenotype.

In vitro response to mitogens and antigens in elderly and post-cytotoxic patients can be modified by a new hypoxanthine derivative (PCF-39).

The in vitro immunomodulatory properties of the hypoxanthine derivative N alpha-5(1.6-dihidro-6-oxo-9-purinyl)pentyloxy/carbonyl -L-arginine (PCF-39) are described. Data obtained with PHA in vitro indicate that PCF-39 exhibits immunopharmacologic activity in elderly and post-cytotoxic patients, while showing no significant activity in normal donors. Antigen induced proliferation was unaffected by PCF-39, while PMN induced chemiluminescence was significantly enhanced. PCF-39 can be considered a new drug in the family of the hypoxanthine derivatives endowed with immunomdulatory properties.

Sensory neuropeptides (substance P) and 4-11 SP enhance human neutrophils chemiluminescence; the role of L-arginine.

Investigations on the effect of a naturally occurring neuropeptide, substance P (SP) and one of its synthetic analogues, 4-11 SP, on luminol-enhanced chemiluminescence (CL) of human polymorphonuclear leukocytes (PMN) are presented. Both peptides elicited a strong burst of CL with different time course and dose-response curves. SP- and tuftsin-induced CL were similar, both peptides sharing a Lys-Pro-Arg terminal; 4-11 SP, which lacks the terminal arginine, peaked earlier than the natural peptide. Surprisingly when PMN were pre-incubated with L-arginine (L-Arg) this enhancing effect was abolished or diminished. L-Arg and an L-Arg synthetic derivative (PCF-39) were also evaluated; PCF-39 strongly increased the PMN CL, while L-arginine showed a significant CL enhancement only in 43% of the donors tested. Peptide- and arginine-induced CL were Ca++/MG++ dependent.

[Neuroblastoma at the Giannina Gaslini Institute. Analysis of 94 cases diagnosed 1972-1981]. / Neuroblastoma all'Istituto Giannina Gaslini. Analisi di 94 casi diagnosticati nel periodo 1972-1981.

The authors have analysed 94 consecutive previously untreated children affected by NB diagnosed at the Giannina Gaslini Children's Hospital in the period January 1972 - December 1981. Age at diagnosis ranged between 1 month - 16 years (median 2 years). Diagnosis was made on histological grounds in 82 cases, on clinical, instrumental and laboratory data in the remaining 12 cases. Evans' staging system was adopted for classifying the disease extent. Patient recruitment was 9.4 cases for year. Female sex slightly prevailed in our series. 17 patients had localized disease (stage I and II), all surviving since 27 - 90 months. 28 children had regional disease (stage III) half of whom are presently alive. Five of the 47 cases with disseminated disease are alive at the time of this study; only one of these 5 can be considered cured, having been followed since more than 4 years. One of the two IV-s stage children survives disease-free at 89 months. 49% of patients were 2-year old or less at diagnosis while only 2 patients were older than 10. Survival was best in children diagnosed under one year of age, lowest in the 2 - 6 year age group. 74% of patients had their primary located in the abdomen (36 in the adrenal, 16 in sympathetic ganglia, 18 in un unidentifable site). Prognosis was worse in these patients compared with those with primary in the thorax, neck and pelvis. The clinical presentation of the disease has been extremely various: most patients had a diagnosis different from the definitive one, and were accordingly treated usually for several weeks or even months. Among laboratory data, urinary catecholamine metabolites, serum LDH and serum ferritin represented the most suitable indices of disease activity. Among immunological studies, none has shown a good correlation with the extent of the disease, nor with the clinical course. Stage I children had only surgery as treatment, while in stage II surgical ablation was followed by short-lasting chemotherapy and radiotherapy in few cases. In stage III and IV more complex multidisciplinary approach was utilized in the attempt to achieve a complete tumor regression. While about a half stage III cases appears curable by these treatment modalities, little impact has been demonstrated by this therapy in widespread disease, although the use of well-designed protocols seems have improved the complete remission rate and the median length of survival.