bluuebunny/crossref_metadata_2025 huggingface dataset:
Granulocyte‐macrophage colony‐stimulating factor stimulates the metastatic properties of lewis lung carcinoma cells through a protein kinase a signal‐transduction pathway
Authors: Young M. Rita I. , Lozano Yvonne , Djordjevic Andelka , Devata Sandeep , Matthews John , Young Melvin E. , Wright Mark A.
Published: February 2007

Expression of granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) by metastatic Lewis lung carcinoma cells (LLC‐ LN7) was previously shown to contribute to the maintenance of phenotypic characteristics associated with an increased capacity to metastasize. In the present study, pre‐incubation of LLC‐LN7 cells with neutralizing anti‐GM‐CSF antibodies diminished the capacity of the tumor cells to form experimental metastases after i.v. inoculation, while pre‐incubation with recombinant GM‐CSF (rGM‐CSF) increased formation of metastases. In the presence of rGM‐CSF, the LLC‐LN7 cells exhibited an increased capacity to migrate, invade through a reconstituted basement membrane, and adhere to lung tissue. Studies to identify the signal transduction pathway through which GM‐CSF enhanced the in vitro metastatic properties of the LLC‐LN7 tumor cells implicated protein kinase A (PKA). Signaling through PKA was suggested by the demonstration that the stimulation of tumor‐cell motility by GM‐CSF was blocked in the presence of the adenylate cyclase inhibitor nicotinic acid, or the PKA inhibitors A3 or KT5720. In addition, the role of PKA as a signaling mechanism for GM‐CSF was assessed by using REV‐LN7 cells, which are LLC‐LN7 cells that have been stably tranrfected with an expression vector encoding a mutant PKA RIα. subunit and which, in turn, express a cAMP‐resistant PKA. Adherence and invasion by the PKA‐defective REV‐LN7 cells were not stimulated by rGM‐CSF, contrasting with the stimulation observed for wild‐type LLC‐LN7 cells. These data suggest that rGM‐CSF can further enhance the in vitro metastatic characteristics of LLC‐LN7 tumor cells and that this is dependent on signal transduction through PKA.
Generation of cytotoxic antibodies to the B16 murine melanoma using a formalinized vaccine
Authors: Shrayer David , Kouttab Nicolas , Maizel Abby , Wanebo Harold , Hearing Vincent J. , Gersten Douglas M.
Published: February 2007

The goal of our experiments was to determine the extent to which the humoral response to a melanoma vaccine elicits the production of cytotoxic antibodies in tumor‐challenged mice. Mice were immunized with a vaccine produced from formalinized extracellular antigens (FECA) derived from B 16 F 10 melanomas. The production of antibodies that recognized the vaccine preparation was determined by ELISA, as was their cross‐reactivity with the B700 melanoma,antigen. The antibodies were shown to be anti‐proliferative by inhibition of tritiated thymidine incorporation into the DNA of cultured target cells and cytotoxic by assays for complement‐mediated and antibodydependent cellular cytotoxicity. Flow cytometric analyses indicated that‐60% of the target cells specifically bound antibody from the immune sera. These results confirm that B700 is a significant antigenic component of the FECA vaccine, and provide encouragement for this approach to developing useful melanoma vaccines.
Enhancement by isoproterenol of hepatocarcinogenesis induced by n‐nitrosomorpholine in sprague‐dawley rats
Authors: Uehara Hiroyuki , Nakaizumi Akihiko , Baba Miyako , Iishi Hiroyasu , Tatsuta Masaharu
Published: February 2007

The effect of isoproterenol on hepatocarcinogenesis induced by N‐nitrosomorpholine (NNM) was investigated in male Sprague‐Dawley rats. Rats were given drinking water containing NNM for 8 weeks and S.C. injections of isoproterenol or vehicle every other day for 13 weeks. Pre‐neoplastic and neoplastic lesions staining positive for γ‐glutamyl transpeptidase (GGT) or the placental type of glutathione‐S‐transferase (GST‐P) were examined histochemically at week 13. Prolonged administration of isoproterenol resulted in a significant increase in the number of GGT‐positive, but not GST‐P‐positive, lesions. The incidence, number and size of hepatocellular carcinomas were also significantly greater in rats treated with isoproterenol than in controls. Administration of isoproterenol significantly increased the intracellular cAMP and the labeling indices of pre‐neoplastic lesions and adjacent liver. These findings indicate that isoproterenol enhances hepatocarcinogenesis and that this may be related to its enhancing effect, mediated by cAMP, on cell proliferation in neoplastic lesions and surrounding hepatocytes.
Food‐group consumption and colon cancer in the adelaide case‐control study. I. Vegetables and fruit
Authors: Steinmetz Kristi A. , Potter John D.
Published: February 2007

Previous epidemiologic studies have shown an inverse association between vegetable and fruit consumption and risk of colon cancer. Vegetables and fruit contain a large number of potentially anti‐carcinogenic substances, thus lending biological plausibility to this association. We conducted a case‐control study in Australia, comparing 220 persons with histologically confirmed incident adenocarcinoma of the colon with 438 age‐ and gender‐matched controls. Cases were identified via the South Australian Cancer Registry (1979‐80); controls were randomly selected from the electoral roll. All participants completed 14I‐item food‐frequency questionnaire and were interviewed regarding demographic and other information. Consumption of 15 vegetable and fruit groups was investigated. Odds ratios (OR) for quartiles of consumption were derived using conditional logistic regression. All analyses were conducted separately for females and males. For females, greater intakes of onions and legumes were associated with decreased risk, with protein‐adjusted OR of 0.48 and 0.53 respectively. Greater intakes of raw fruit and cabbage were associated with protein‐adjusted OR of 0.76 and 0.71 respectively. For males, greater intakes of onions, green leafy vegetables, legumes, carrots and cabbage were associated with protein‐adjusted OR in the range of 0.72 to 0.77. Consumption of potatoes was positively associated with risk in both genders. All 95% confidence intervals included I.O. Analyses stratified by colon‐cancer sub‐site showed no strong and consistent differences between sub‐sites for the vegetable and fruit associations. Results for meat, poultry, seafood, dairy foods and eggs are presented in a companion report.
Food‐group consumption and colon cancer in the adelaide case‐control study. II. Meat, poultry, seafood, dairy foods and eggs
Authors: Steinmetz Kristi A. , Potter John D.
Published: February 2007

We conducted a case‐control study in Australia, comparing 220 persons with histologically confirmed incident adenocarcinoma of the colon with 438 age‐ and gender‐matched controls. Cases were identified via the South Australian Cancer Registry (1979‐80); controls were randomly selected from the electoral roll. All participants completed a 14I‐item food‐frequency questionnaire and were interviewed regarding demographic and other information. Consumption of 8 groups of foods from animal sources was investigated. Odds ratios (OR) for quartiles of consumption were obtained using conditional logistic regression. All analyses were conducted separately for females and males. The most striking finding was a positive association for egg consumption in females, with an unadjusted OR of 2.4 (1.1‐5.3) for consumption in the uppermost quartile. The uppermost septile of egg consumption was associated with an unadjusted OR of 6.3 (1.5‐26.1) and a dose‐response pattern was suggested. Intakes of red meat, liver, seafood, and dairy foods were also weakly positively associated with risk in females. In males, intakes of red meat and poultry were weakly positively associated with risk with unadjusted ORs of 1.5 (0.8‐2.8) and 1.4 (0.7‐2.6) respectively. The ratio of intake of red meat to poultry and seafood was also positively associated with risk in males, with an unadjusted OR of 1.4 (0.8‐2.6). Interpretation of analyses stratified by colon cancer subsite was limited by the low number of subjects in each sub‐site stratum, yet the results were somewhat supportive of a stronger risk associated with animal foods in the proximal than in the distal colon. The results for egg consumption suggest a role for cholesterol in the etiology of colon cancer, particularly in proximal cancer for females. Results for vegetable and fruit consumption are presented in a companion report.
Cancer risk in farmers: Results from a multi‐site case‐control study in north‐eastern italy
Authors: Franceschi Silvia , Barbone Fabio , Bidoli Ettore , Guarneri Salvatore , Serraino Diego , Talamini Renato , La Vecchia Carlo
Published: February 2007

The relationship between farming and cancer risk was investigated in an integrated series of case‐control studies conducted from 1985 to 1991 in the Friuli Venezia Giulia region, north‐east Italy. Patients with cancer of the oral cavity and pharynx, larynx, oesophagus, colon and rectum, pancreas, breast, thyroid gland, kidney and urinary tract, bladder, prostate, soft‐tissue sarcomas, Hodgkin's diseases, non‐Hodgkin's lymphomas and multiple myelomas, and controls admitted to hospital for acute, non‐neoplastic conditions, were interviewed. For males, a significantly elevated relative risk was seen for oral cavity and pharynx. Farming, however, was associated with a significant protection against cancer of the colon and rectum and bladder. In females, only one significant association emerged, for multiple myeloma. A few significant interactions between cancer risk and year of birth (i.e., before 1930 or 1930 and after) were observed. The risk of cancer of the larynx was significantly elevated in younger male farmers but not in older ones. Our multi‐site case‐control study confirms that farmers have, for some cancer sites, a distinctive pattern. Excesses of cancer of the oral cavity and pharynx in farmers are characteristic of the present study area and, possibly, of similar European rural populations who have in common high levels of alcohol consumption and, at least in the past, unbalanced diets.
Epstein‐barr virus carriage by nasopharyngeal carcinoma in situ
Authors: Yeung W. M. , Zong Y. S. , Chiu C. T. , Chan K. H. , Sham Jonathan S. T. , Choy Damon T. K. , Ng M. H.
Published: February 2007

We studied the distribution of the EBV genome in tumour biopsies obtained from 42 patients with poorly differentiated or undifferentiated nasopharyngeal carcinoma (NPC) and 3 patients with well‐differentiated NPC. Six carcinoma in situ (CIS) foci were seen in 5 tumour specimens. By in‐situ hybridization, multiple copies of the EBV genome were detected in some of the tumour cells in 3 CIS lesions involving the full thickness of the mucosal epithelium, but without microinvasion, while the viral genome was present in the majority of the tumour cells contained in another 3 CIS lesions with microinvasion. In agreement with previous findings, poorly differentiated and undifferentiated carcinomas regularly carried the viral genome, the number of copies of which was similar to that seen in CIS, while some, but not all, of the tumour cells of the welldifferentiated histological type carried the virus. The viral genome was otherwise rarely detected in other areas of the mucosal epithelium and, where present, the viral carriage was confined to a few epithelial cells, in which the viral genome contents were markedly lower than in tumour cells. These results suggest that EBV may first become associated with NPC at an early stage of the disease shortly after the tumour has been initiated.
CA 494—a new tumor marker for the diagnosis of pancreatic cancer
Authors: Friess Helmut , Büchler Markus , Auerbach Bernhard , Weber Alexander , Malfertheiner Peter , Hammer Kai , Madry Norbert , Greiner Sabine , Bosslet Klaus , Beger Hans G.
Published: February 2007

In 59 patients with ductal pancreatic cancer the monoclonal antibody (MAb) BW 494, which detects the CA 494 glycoprotein antigen, was analyzed in comparison with the reference tumor markers CA 19‐9 and CEA. Eighty‐one patients with non‐pancreatic malignancies of the gastraintestinal (GI) tract, 95 with chronic pancreatitis, 124 with benign non‐pancreatic GI diseases, 30 with diabetes mellitus (type I or type II) and I14 healthy blood donors served as controls. The sensitivity of pancreatic cancer was 90%, 44% and 90% for CA 19‐9, CEA and CA 494, respectively. In chronic pancreatitis, as the most important control population for pancreatic cancer, the specificity was 85%, 72% and 94% for CA 19‐9, CEA and CA 494, respectively.
Loss of RB protein expression in primary bladder cancer correlates with loss of heterozygosity at the RB locus and tumor progression
Authors: Xu Hong‐Ji , Cairns Paul , Hu Shi‐Xue , Knowles Margaret A. , Benedict William F.
Published: February 2007

RB‐protein status as determined by immunohistochemical analysis was compared with loss of heterozygosity (LOH) at the RB locus in 68 primary transitional‐cell carcinomas of the bladder. Absence of RB‐protein expression was found in 15 of 17 tumors in which LOH at the RB locus was identified, whereas 31 of 36 tumors from informative patients which showed no LOH had a normal RB‐protein pattern ( p < 0.001). Altered RB‐protein expression was also more frequently seen in muscle‐invasive and high‐grade tumors ( p < 0.003 and <0.005, respectively). Our results indicate that LOH at the RB locus is highly correlated with loss of RB‐protein expression in primary bladder carcinomas and further strengthens the notion that loss of RB function may be associated with more aggressive bladder tumors.
Inhibition by dehydroepiandrosterone of butylated hydroxyanisole (BHA) promotion of rat‐bladder carcinogenesis and enhancement of BHA‐induced forestomach hyperplasia
Authors: Shibata Masa‐Aki , Shirai Tomoyuki , Asakawa Emiko , Hirose Masao , Fukushima Shoji
Published: February 2007

The effects of dehydroepiandrosterone (DHEA) with/without ribonucleoside (RNs) supplementation on butylated hydroxyanisole (BHA) bladder‐tumor promotion and forestomach carcinogenesis were investigated. Male F344 rats were given N‐butyl‐N‐(4‐hydroxybutyl)nitrosamine (BBN) in their drinking water for 4 weeks and then received basal diet or diet containing BHA, DHEA, a mixture of RNs, BHA + DHEA or BHA + DHEA + RNs for 32 weeks. The occurrences of papillomas and carcinomas in the urinary bladder were increased in the groups given BHA or BHA + DHEA + RNs, as compared with control group values. In comparison with the BHA group, the BHA + DHEA group incidences and numbers of these tumors were decreased. However, the incidence and multiplicity of papillomas in the group given BHA + DHEA + RNs were again elevated. DNA synthesis levels in normal‐appearing bladder epithelium, but not tumor cells, were closely correlated with the observed level of promotion in most groups. The case of DHEA alone proved exceptional in that DNA synthesis was markedly decreased without any significant influence on lesion development. In the forestomach, DHEA, which itself was associated with slight although non‐significant hyperplasia, enhanced BHA‐induced epithelial lesions, characterized by marked basal‐cell proliferation and keratin‐cyst formation, independently of additional RNs administration.

Our results suggest that the anti‐promoting effects of DHEA in the bladder depend on a deficiency in the pentose phosphates necessary for production of nucleosides. Organ‐specific modulation is indicated by the enhancing effects of DHEA on BHA‐induced forestomach hyperplasia.
Binding of some metastatic tumor cell lines to fibrous elastin and elastin peptides
Authors: Svitkina Tatyana M. , Parsons Donald F.
Published: February 2007

Recent suggestions that tumor‐cell targeting of elastin‐rich tissues (e.g., lung) correlates with the presence of surface elastin receptors have been investigated. Receptors for insoluble (fibrous) elastin and for soluble elastin peptides have been implemented in these correlations. A rapid assay for binding of insoluble elastin has been devised. Two of the cell lines tested (M27 and MAT‐LyLu), which metastasize to the lung, strongly bound fibrous elastin whereas a third (B16‐F10) did not. None of 4 metastatic cell lines that do not target the lung (A549,3LL, TA3, TA3‐iso2) bound fibrous elastin. The ability of cell lines to interact with soluble elastin was tested by cell attachment to high‐molecular‐weight soluble elastin peptides adsorbed on plastic surface. Three of 7 tested cell lines, B16‐F10, M27 and TA3, attached to a soluble elastin coating. In contrast to the rapid binding of insoluble elastin particles, the cell interaction with immobilized soluble elastin peptides was delayed, suggesting that induction of receptors for soluble elastin and/or modification of the elastin coat was occurring. Thus, all 3 tested cell lines where metastases target the lung, namely, MAT‐LyLu, B16‐F10 and M‐27, show soluble‐ or insoluble‐elastin interactions, whereas, of 4 cell lines not targeting lungs, only one, TA3, reacts with soluble elastin.
Cell culture of tumors alters endogenous poly(ADPR)polymerase expression and activity
Authors: Chabert Magali G. , Kopp Patrick C. , Bischoff Pierre L. , Mandel Paul
Published: February 2007

Poly(ADP‐ribose)polymerase, a chromatin‐bound enzyme, actively participates in processes such as cell proliferation, differentiation, and DNA repair and replication. This enzyme is also implicated in cell transformation, and its inhibition has been proposed to potentiate anti‐cancer drug activity. Since cells prepared from tumor biopsies and established tumor cell lines are commonly used to evaluate the efficiency of anticancer therapies, we have compared poly(ADP‐ribose)polymerase activity in animal tumor cells growing in vivo and in cell culture. Three tumor types were tested: a mastocytoma (P815), a lymphoma (RDM4). and a glioma (C6). Our results show that cell culture alters poly(ADP‐ribose)polymerase levels and activity. Endogenous poly(ADP‐ribose) activity was several fold higher in exponentially growing cells than in cells freshly recovered from solid or ascitic tumors. Moreover, polymerase activity increased with culture time, reaching a maximum when cells became confluent. Measurements of poly(ADP‐ribose)polymerase gene expression and protein amount indicate that lower enzyme activity in tumors grown in vivo are sustained by decreases in poly(ADP‐ribose)polymerase mRNA and protein amount. In contrast, the increase in endogenous poly(ADP‐ribose)polymerase activity observed in cultured cells was due to enzyme activation and not to de novo protein synthesis. Such differences must be considered when assessing the applicability of cell‐culture results to in vivo situations.
Insulin dependence of murine T‐cell lymphoma. II. Insulin‐deficient diabetic mice and mice fed low‐energy diet develop resistance to lymphoma growth
Authors: Sharon Raphael , Pillemer Graciela , Ish‐Shalom Dvorah , Kalman Rony , Ziv Ehud , Berry Elliot M. , Naor David
Published: February 2007

Physiological concentrations of insulin support the in vitro growth of LB T‐cell lymphoma. We could not detect similar insulin dependence in other tumor cell lines. This study reports that insulin also enhances the growth of LB cells in vivo. Mice treated with Streptozotocin (SZ) developed partial resistance to LB lymphoma growth and they survived longer (p < 0.0025) than non‐diabetic mice after LB‐cell inoculation. A few diabetic mice developed complete tumor resistance, manifested by total regression of the lymphoma. SZ‐treated diabetic mice reconstituted with external insulin died as fast as non‐diabetic mice when both were inoculated with the same number of LB cells. The SZ‐treated diabetic mice did not develop resistance to the growth of BCLI B‐cell leukemia, which demonstrated only a marginal proliferative response to insulin in vitro. Mice fed a low‐energy diet exhibited low insulin levels and also developed resistance to lymphoma growth (50% survival 21 days vs. 15 days; p < 0.0005), supporting the concept that insulin enhances LB T‐cell tumor development in mice.
Expression of integrins and other adhesion molecules on NK cells; Impact of IL‐2 on short‐ and long‐term cultures
Authors: Mäenpää Anna , Jääskeläinen Juha , Carpén Olli , Patarroyo Manuel , Timonen Tuomo
Published: February 2007

We have investigated, using flow cytometry, the expression of 19 adhesion molecules on fresh and IL‐2‐activated NK cells. The study included β1, β2 and β3 integrins, CD2, CD54 and CD58 (belonging to the immunoglobulin superfamily), and CD44 and L‐selectin (homing receptors). α1 and α2 of the β1 integrins were non‐existant and α3 was weak on freshly isolated NK cells, but their expression increased after 4 weeks in culture with IL‐2. On the other hand, some down‐regulation of α4 and α5 and disappearance of α6 was detected. CD11a/CD18 was upregulated by IL‐2, whereas CD11b‐c/CD18 were down‐regulated. As a novel finding we detected β3 on IL‐2‐activated T and NK cells. CD2, CD44, CD54 and CD58 were increased by IL‐2 but L‐selectin was strongly down‐regulated on the long‐term‐activated NK cells. Although IL‐2‐activated lymphocytes are potent tumor‐lysing killer cells in vitro and therefore a potential modality in cancer treatment, the IL‐2 induced changes in lymphocyte adhesion molecule expression may also lead to undesired effects, such as altered untargeted distribution and compromised migratory capacity.
Camptothecin derivatives induce regression of human ovarian carcinomas grown in nude mice and distinguish between non‐tumorigenic and tumorigenic cells in vitro
Authors: Pantazis Panayotis , Kozielski Anthony J. , Mendoza John T. , Early Janet A. , Hinz Hellmuth R. , Giovanella Beppino C.
Published: February 2007

We have recently shown that the plant alkaloid 20(S)‐camptothecin and its derivatives 9‐nitro‐20(S)‐camptothecin(9NC) and 9‐amino‐2O(S)‐camptothecin(9AC) inhibit the growth of a variety of human tumors xenografted in nude mice. In this report, we demonstrate that 9NC and 9AC effectively inhibit growth, and subsequently induce regression, of human ovarian tumors grown in nude mice. Tumor regression is accompanied by degenerative changes in the tumor cells as assessed by microscopic observations of histological sections prepared from the tumors. Parallel experiments in vitro show that 9NC inhibits in a similar manner the growth of human ovarian carcinoma cells, regardless of their ability to induce tumors when xenografted in nude mice, and induces similar morphological changes in both non‐tumorigenic and tumorigenic cells, as assessed by microscopic observation. Flow cytometry studies show that 9NC‐induced growth inhibition of the non‐tumorigenic cells is associated with accumulation of these cells in G2. In contrast, 9NC‐induced growth inhibition of the tumorigenic cells is associated with the generation of cells containing a reduced DNA content, that is, cells programmed to die. In conclusion, camptothecins appear to be cytostatic for non‐tumorigenic, but cytotoxic for tumorigenic cells, an important finding from viewpoints of cell biology, pharmacology and cancer chemotherapy.
Immortalization of human cervical keratinocytes by human papillomavirus type 33
Authors: Gilles Christine , Piette Jacques , Rombouts Sonia , Laurent Christian , Foidart Jean‐Michel
Published: February 2007

Ten immortalized cell lines were established by transfection of human cervical keratinocytes (CK) with HPV‐33 DNA and some of their characteristics were investigated. The following observations were made: (a) several cell lines have reached over 100 population doublings in vitro; (b) 3 transcripts were observed, 2 being encoded by the E6/E7 open reading frames (ORFs); (c) cytogenetic analyses showed important genetic modifications such as aneuploidy and isochromosome formation of the q arm of chromosome 8; (d) 2 of the 10 cell lines developed colonies in soft agar but none was able to form tumors when injected S.C. into nude mice; (e) Southern analysis suggested that a single copy of HPV‐33 is integrated at a single common site within the genome of the 10 cell lines. These immortalized cell lines should be useful for studying mechanisms involved in proliferation, differentiation and neoplastic transformation of CK by HPV‐33.
In vitro anti‐tumor activity of eosinophils from cancer patients treated with subcutaneous administration of interleukin 2. Role of interleukin 5
Authors: Rivoltini Licia , Colombo Mario Paolo , Parmiani Giorgio , Viggiano Vincenzo , Spinazzè Silvia , Santoro Armando , Takatsu Kiyoshi
Published: February 2007

Interleukin 2 (IL‐2) administration is known to induce marked eosinophilia. To evaluate the potential role of eosinophils as anti‐tumor effectors and to understand the direct or indirect effects of IL‐2 on eosinophils, the physical and functional characteristics of eosinophils obtained during IL‐2 therapy were compared with those of eosinophils obtained from the same patients before IL‐2 administration, or from healthy donors. The treatment schedule consisted of subcutaneous (s.c.) injections of IL‐2, and was performed in 7 patients with small‐cell lung cancer (SCLC) in advanced stage. A marked increase of hypodense cells in peripheral blood was found to correlate with eosinophil activation in patients undergoing IL‐2 therapy. Cyto‐toxic activity of eosinophils against allogeneic tumor cells (SCLC, K562 and melanoma lines), as assessed by direct and antibody(Ab)‐dependent cellular cytotoxicity (ADCC), was markedly increased during IL‐2 therapy. Conversely, eosinophils obtained before treatment, like those of healthy donors, lacked any activity against tumor cells. Sera from IL‐2‐treated, but not from untreated, patients, significantly improved the in vitro survival and anti‐tumor cytotoxicity of eosinophils from healthy donors. Comparable effects were obtained with eosinophils cultured with interleukin 5 (IL‐5), granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) and, to a lesser extent, by tumor necrosis factor‐α (TNFα), while no direct activity was mediated by IL‐2. A 91% inhibition of eosinophil ADCC was found after pre‐incubation of the sera of IL‐2‐treated patients with anti‐IL‐5 but not with anti‐GM‐CSF or anti‐TNFα Ab. IL‐5 mRNA expression was detected in peripheral‐blood lymphocytes (PBL) obtained 4 hr after IL‐2 injection during the second and third week of IL‐2 therapy. Phenotypic analysis of eosinophils from IL‐2‐treated patients showed enhanced expression of activation markers, including Fcγ RII (CD32), HLA‐DR, CR3 (CD I lb) and CRI (CD35). These findings suggest that a significant cytotoxicity against tumor cells can be mediated by eosinophils after indirect, IL‐5‐mediated in vivo activation by IL‐2, and that eosinophils may be involved in the anti‐tumor response(s) induced in vivo by IL‐2.
Absence of HTLV‐I infection among seronegative subjects in an endemic area of Japan
Authors: Kinoshita Tomohiro , Imamura Jun , Nagai Hirokazu , Shimotohno Kunitada , Ito Mizuko , Ito Shin‐Ichiro , Ikeda Shuichi , Nagatomo Masazumi , Tajima Kazuo
Published: February 2007

To examine the prevalence of infection by human T‐cell leukemia virus type I (HTLV‐I) among seronegative subjects, healthy subjects on Tsushima Island, Japan, where the infection is endemic, were evaluated. A total of 209 healthy adults were examined for HTLV‐I provirus in peripheral blood mononuclear cells by the polymerase chain reaction (PCR), as well as for anti‐HTLV‐I antibodies by the particle agglutination (PA) method, the enzyme‐linked immunosorbent assay (ELISA) and by immunofluorescence analysis (IF). A total of 76 subjects were positive and 133 were negative for the provirus, showing a close correlation with the results of 3 assays for anti‐HTLV‐I serum antibodies. None of the seronegative subjects reacted positively on PCR analysis. These observations indicate that seronegative HTLV‐I carriers are rare in an area of Japan in which this viral infection is endemic.
High incidence of monoclonal EBV episomes in Hodgkin's disease and anaplastic large‐cell ki‐1‐positive lymphomas in HIV‐1‐positive patients
Authors: Boiocchi Mauro , De Re Valli , Doixetti Riccardo , Marzorro Alessandra , Gloghini Annunziata , Carbone Antonino , Vaccher Emanucla , Bertola Giulio
Published: February 2007

A series of selected lymphoid malignancies (LMs) occurring in Italian HIV‐1‐infected (HIV+) patients, principally intravenous drug users, was investigated. In addition to small non‐cleaved‐cell (SNCC) and large‐cell immunoblastic (LCI) non‐Hodgkin's lymphomas (NHLs), a relatively high occurrence of anaplastic large‐cell Ki‐1‐positive (ALC Ki‐1+) lymphomas and Hodgkin's disease (HD) was observed, at variance with other reported series of HIV+ patients. Combined results of in situ hybridization and Southern‐blot analyses, in conjunction with immunohisto‐chemical detection of Epstein‐Barr virus (EBV)‐encoded latent membrane protein‐1 (LMP‐1), showed an almost complete association of ALC Ki‐1+ lymphomas and HD cases with EBV. The neoplastic cells of both these LMs also showed common immunophenotypic features such as frequent absence of B‐ and T‐cell differentiation markers and expression of the Ki‐1 activation marker, while SNCC and LCI lymphomas were mainly of mature B‐cell origin and Ki‐1‐. The concomitant high incidence of ALC Ki‐1+ lymphomas and HD in a specific group of HIV+ patients, their almost complete association with EBV in clonal and episomal form and the great similarity in differentiation, activation and virological markers which they display suggest that these LMs are pathological variants of a continuous spectrum of HIV‐1‐associated disorders etiopathologically linked to EBV.
Analysis of T‐cell‐receptor variable gene segment usage in peripheral‐blood lymphocytes of advanced cancer patients
Authors: Angevin Eric , Farace Françoise , Genevée Catherine , Caignard Anne , Dietrich Pierre‐Yves , Hercend Thierry , Triebel Frédéric , Henry‐Amar Michel , Escudier Bernard
Published: February 2007

Advanced cancer patients generally display impaired T‐cell immune functions. The underlying mechanisms are not well understood. The aim of this study was to analyze whether major alterations of TCR variable gene segment usage could be detected in the blood of these patients. Seventeen individuals with various malignancies were tested using PCR and a panel of V‐gene‐segment‐sub‐family‐specific (Vα1‐w29/Vβ1‐w24) oligonucleotide primers. The results indicate that these cancer patient lymphocytes expressed most Vα and Vβ sub‐family specificities, similarly to the lymphocytes of healthy donors (n = 10). This suggests that immunodepression in advanced cancer patients is not related to major deletions in their T‐cell repertoires. We also compared the mean relative expression of each V‐sub‐family specificity of patients and healthy donors by quantitative densitometric analysis. We found significant differences in 4 Vβ specificities (and no Vα). Our analysis identified unique T‐cell sub‐sets putatively involved in the mechanisms leading to immunodepression in advanced cancer patients. Alternatively, the observed differences in terms of Vβ specificity expression may reflect the host response against the tumor.
Integrin expression in cutaneous malignant melanoma: Association of the α3/β1 heterodimer with tumor progression
Authors: Natali P. G. , Bartolazzi A. , Cavaliere R. , Bigotti A. , Nicotra M. R.
Published: February 2007

The cell‐surface heterodimers of the integrin family of molecules, which mediate cell‐cell and cell‐substratum interactions, are likely to be functionally relevant in local and metastatic tumor growth. In the present study we have analyzed whether the α3/β1 receptor for collagen, laminin and fibronectin undergoes changes in expression during tumor progression in cutaneous malignant melanoma (CMM). The results of this study have demonstrated that, while low levels of VLA3 expression are detectable in benign lesions, in primary melanomas the heterodimer undergoes progressive increase in expression which correlates with the degree of dermal invasiveness. Metastatic lesions were found VLA3 positive in 82% of cases. Furthermore, the heterodimer is homogeneously expressed in multiple autologous metastases. The presence of VLA3 correlates with detection of at least one of the ligands in 45% of the cases studied. These findings provide additional evidence that tumor progression in CMM is associated with changes in integrin phenotypes which include the α3/β1 heterodimer.
Regulation of urokinase‐type plasminogen activator expression in squamous‐cell carcinoma of the oral cavity
Authors: Clayman G. , Goepfert H. , Boyd D. D. , Wang S. W. , Nicolson G. L. , El‐Naggar A. , Mazar A. , Henkin J. , Blasi F.
Published: February 2007

We undertook a study to determine the role and regulation of expression of urokinase‐type plasminogen activator in squamous‐cell carcinoma of the oral cavity. The contribution of urokinase to the invasive process was clearly demonstrated in experiments in which in vitro invasion by a cultured squamous‐cell carcinoma cell line was substantially reduced by a monoclonal antibody directed at the catalytic site of urokinase. The conditioned medium from 2 invasive cell lines contained high levels of urokinase. Examination of resected tumors for urokinase revealed (a) localization of the antigen to the tumor cells and (b) higher levels of the plasminogen activator in tumor tissue than in adjacent non‐malignant tissue. These results suggested that elevated expression of urokinase is a common feature of this malignancy. The mRNA half‐lives of cell lines expressing high and low levels of urokinase were similar, indicating that elevated levels of mRNA for the plasminogen activator were not a consequence of a more stable transcript. No evidence of u‐PA gene amplification was obtained by Southern blotting of DNA derived from the cell lines expressing high urokinase levels. Transfection of squamous‐cell carcinoma cells, which express high levels of urokinase, with a construct bearing the chloram‐phenicol acetyl transferase gene driven by the full‐length (2345 bp) urokinase promoter indicated activation of the urokinase promoter in trans. In conclusion, our results suggest that transcriptional activation of the urokinase gene, in squamous‐cell carcinomas of the oral cavity, leads to elevated levels of urokinase mRNA/protein which, in turn, can promote the invasive phenotype.
Allelic loss on chromosome 17 in human ovarian cancer
Authors: Phillips Nancy , Ziegler Michelle , Saha Bratin , Xynos Francisco
Published: February 2007

In order to identify a common region of deletion on chromosome 17 potentially containing a tumor‐suppressor gene, 27 ovarian carcinomas and 3 ovarian tumors of low malignant potential (LMP) were examined for loss of heterozygosity (LOH) at 6 p arm and 10 q arm loci. Ninety percent of all tumors had deletions at one or more loci. On the p arm, there was a single near‐common region of deletion on 17p 13.3 (D/7S30/ pYNZ22.1; 86% LOH), an intervening locus with a low LOH rate, and a more proximal locus on 17p11.2 (D/7S58/pEW301; 82% LOH) with a high LOH rate. In less aggressive tumors, LOH at Df 7S30 was not accompanied by LOH at p53. The q arm had a common region of deletion for high‐stage carcinoma at D/7S579 (Mfd 188; 74% LOH) on q21, a locus tightly linked to the familial breast‐ovarian‐cancer syndrome (BRCAI) locus. D/7S579 was lost in all informative high‐stage carcinomas and retained in all low‐stage carcinomas and tumors of LMP. There may be at least 2 tumor‐suppressor genes, an early‐acting gene on the p arm and a gene on the q arm involved in tumor progression and metastasis.
Proliferation, migration and invasion of human glioma cells exposed to antifolate drugs
Authors: Terzis A. J. A. , Arnold H. , Fiskerstrand T. , Refsum H. , Ueland P. M. , Bjerkvig R.
Published: February 2007

The present study describes the effects of 2 folate antagonists, methotrexate (MTX) and the lipophilic antifolate trimetrexate (TMX) on 2 permanent human glioma cell lines (GaMg and D‐54Mg) grown as monolayers and as multicellular tumor spheroids. In addition, the effects of drug exposure on tumor cell invasion was studied using a three‐dimensional organ co‐culture system. In monolayer cultures, TMX was a more potent inhibitor of cell growth than MTX, especially towards the GaMg cell line. The 2 drugs, however, showed similar cytotoxicity as assessed by the plating efficiency assay. Reduced ability of directional migration of cells on a plastic surface was seen by either antifolate usually at concentrations to 10‐fold higher than those exerting a cytotoxic effect in the plating efficiency assay. TMX was somewhat more potent than MTX as an inhibitor of spheroid growth. When tumor spheroids were exposed to MTX or TMX at concentrations that caused 65 to 70% inhibition of cell migration, there was a latent period of 4 to 5 days before inhibition of spheroid growth ensued. Invasion was investigated in a co‐culture system, where tumor spheroids were confronted with fetal rat brain cell aggregates. Neither drug reduced tumor cell invasion, although histological examination revealed toxic effects both in GaMg and in D‐54Mg spheroids. We conclude that spheroids from human glioma cells were less sensitive to the antifolates than monolayers. For both drugs a latency period was observed before inhibition of spheroid growth. The spheroids retained their ability to invade normal brain tissue when exposed to levels of folate antagonists inhibiting spheroid growth.
TGF‐β induces an inhibitory effect of normal cells directed against transformed cells
Authors: Höfler P. , Wehrle I. , Bauer G.
Published: February 2007

Co‐cultivation of normal and transformed fibroblasts in the presence of exogenously added transforming growth factor type beta 1 (TGF‐β1) leads to a drastic reduction of colony‐forming transformed cells. This inhibitory effect is dependent both on the dose of TGF‐β applied and the density of normal cells. It does not reflect a direct action of TGF‐β on transformed cells, but is rather induced by TGF‐β in normal cells. In the absence of exogenously added TGF‐β, normal cells exhibit a basal inhibitory activity, which can be abrogated by anti‐TGF‐β. TGF‐β‐mediated inhibition of transformed cells by normal cells may be considered as a novel potent controlling step in carcinogenesis.
Abrogation of tumor‐inhibitory MRC‐OX8+ (CD8+) effector T‐cell generation in rats by selective depletion of neutrophils In vivo using a monoclonal antibody
Authors: Tanaka Eiichi , Sendo Fujiro
Published: February 2007

These studies were designed to examine the role of neutrophils in transplantation immunity to syngeneic rat tumors. We have earlier reported that specific transplantation immunity to syngeneic transplanted tumors is abrogated by selective depletion of peripheral‐blood neutrophils by administration of a monoclonal antibody (MAb) (RP‐3) at the time of immunization with X‐irradiated tumor cells. In order to elucidate the mechanisms of this phenomenon, we have now examined whether induction of the sensitized spleen cells that inhibit the growth of tumor cells is inhibited by RP‐3 treatment at the time of in vivo priming with tumor‐associated antigen (TAA). Neutrophils were required to induce the sensitized T cells responsible for tumor inhibition with Winn's assay. In addition, CD8+ T cells are proved to be effector cells in an assay system of tumor‐growth inhibition using a diffusion chamber. Our results indicate that neutrophils are required for priming rats with TAA to induce CD8+ effector T cells in tumor inhibition.
Expression of myc family oncoproteins in small‐cell lung‐cancer cell lines and xenografts
Authors: Rygaard Kåre , Spang‐Thomsen Mogens , Vindelø Lars L.
Published: February 2007

A number of genes have altered activity in small‐cell lung cancer (SCLC), but especially genes of the myc family (c‐myc, L‐myc and N‐myc) are expressed at high levels in SCLC. Most studies have explored expression at the mRNA level, whereas studies of myc family oncoprotein expression are sparse. We examined the expression of myc proto‐oncogenes at the mRNA and protein level in 23 cell lines or xenografts. In the cell lines, the doubling time and the cell‐cycle distribution, as determined by flow‐cytometric DNA analysis, were examined to establish whether the level of myc‐gene‐family expression correlated with proliferative parameters. All tumours expressed at least one myc family member at the mRNA level. Exclusive c‐myc mRNA expression was demonstrated in 8 tumours, L‐myc in 7 and N‐myc in I. Five tumours expressed both c‐myc and L‐myc, and 2 tumours expressed both c‐myc and N‐myc. In general, the level of expression of c‐myc and N‐myc was similar at the mRNA and the protein level. Expression of c‐myc was positively correlated with the proliferative index (sum of S and G2 + M phases) of cell lines, but not with the population doubling time. In general, L‐myc‐expressing cell lines had a low proliferative index. There was no systematic difference in myc expression between cell lines and xenografts of individual tumours.
Relationship between the anti‐HTLV‐1 antibody level, the number of abnormal lymphocytes and the viral‐genome dose in HTLV‐1‐infected individuals
Authors: Shinzato Osamu , Shiku Hiroshi , Nagata Yasuhiko , Nakayama Eiichi , Kamihira Shimeru , Ikeda Shuichi , Kondo Hisayoshi , Kanda Takcnao
Published: February 2007

Two distinct diseases, adult T‐cell leukemia (ATL) and HTLV‐1‐associated myelopathy/tropical spastic paraparesis (HAM/TSP), develop in a minor population of HTLV‐1 carriers. We examined the relationship between the viral genome dose in the peripheral‐blood mononuclear cells and the serological response in HTLV‐1 carriers and patients with HAM/TSP. The antibody titer to HTLV‐1 gag and env proteins, as well as the frequency of an antibody response to viral protein p40tax and the titer, increased with increasing viral genome dose. However, the number of abnormal lymphocytes was not directly related to the host viral load. Patients with HAM/TSP generally showed a higher genome dose than healthy carriers and also had higher antibody titers than healthy carriers with the same HTLV‐1 load, supporting the existence of an augmented immune response in these patients. These findings suggest that the antibody titer to HTLV‐1 genome products, and not the number of abnormal lymphocytes, intimately reflects the approximate viral load in HTLV‐I ‐infected individuals.
Cell proliferation as a long‐term prognostic factor in diffuse large‐cell lymphomas
Authors: Silvestrini Rosella , Costa Aurora , Boracchi Patrizia , Giardini Roberto , Rilke Franco
Published: February 2007

The relevance of cell proliferation rate—defined as the 3H‐thymidine labeling index (3H‐dT LI)—in predicting response to treatment (complete remission, CR), freedom from progression (FFP) and overall survival (OS) was evaluated in 86 patients with diffuse large‐cell lymphoma (DLCL). The biologic variable was not associated with most of the established clinical factors, such as gender and age of the patient, performance status, B symptoms, tumor bulk, or extranodal disease, but was directly related to stage. 3H‐dT LI significantly predicted short‐ and long‐term clinical outcome. In fact, more patients with slowly proliferating DLCL reached CR and had longer median FFP and OS than patients with rapidly proliferating DLCL. Multiple‐regression analysis to evaluate the relative contribution of the different biologic and clinical variables in predicting CR, FFP and OS showed that 3H‐dT LI and Ann Arbor stage were the only 2 stable factors, which retained their prognostic significance even in the presence of other conventional factors, and that 3H‐dT LI was the most powerful as an indicator of risk of death in DLCL patients.
Oral contraceptives and endometrial cancer: Do other risk factors modify the association?
Authors: Stanford Janet L. , Brinton Louise A. , Hoover Robert N. , Berman Michael L. , Mortel Rodrigue , Twiggs Leo B. , Barrett Rolland J. , Wilbanks George D.
Published: February 2007

The joint effect of use of combination‐type oral contraceptives and other exposure factors on risk of endometrial cancer was examined in data from a multicenter case‐control study conducted in 5 areas of the United States. Cases were 405 women with histologically confirmed invasive epithelial endometrial cancer first treated at one of 7 participating hospitals. A total of 297 population‐based controls of similar age, race, and geographic area were selected as a comparison group. Information on exposure factors was derived from in‐person interviews. Combination‐type oral contraceptive (COC) use was associated with a significant reduction in risk of endometrial cancer, with an adjusted odds ratio (OR) of 0.4 (95% confidence interval 0.3 to 0.7) for ever compared to never use. Long‐term (≥ 10 years) users experienced a markedly lower risk (OR = 0.2). Women who discontinued COC use ≥20 years earlier remained at reduced risk (OR = 0.7) compared with non‐users. The negative association with COC use was apparent regardless of the presence or level of several other risk factors for endometrial cancer, including age, menopausal status, parity, obesity, ever‐use of menopausal estrogens, smoking history, or history of infertility. The magnitude of the negative association observed in COC users, however, was considerably diminished in women with no full‐term births and in women who subsequently used replacement estrogens for 3 or more years. These results provide new evidence that the protective effect of COC use lasts for 20 or more years after use is discontinued, and highlight several sub‐groups of users in whom the level of protection is attenuated by the presence of other risk factors for this disease.
HTLV‐I and HTLV‐II world‐wide distribution: Reanalysis of 4,832 immunoblot results
Authors: Madeleine Margaret M. , Wiktor Stefan Z. , Goedert James J. , Manns Angela , Levine Paul H. , Biggar Robert J. , Blattner William A.
Published: February 2007

Mapping the geographic distribution of human T‐lymphotro‐phic virus types 1 and 2 (HTLV‐I and ‐II) has been complicated because conventional serologic approaches cannot distinguish between these 2 viruses. To more precisely define the epidemiology of HTLV‐I and HTLV‐II, we evaluated a convenience sample of 4,832 HTLV‐I immunoblot results from over 140,000 samples screened for HTLV‐I, collected in 16 countries for 35 separate studies. An algorithm that compares reactivity against p19 and p24, 2 gag (HTLV‐I core) proteins, was employed to characterize the immunoblots: type I, p19 stronger than p24 (presumptive HTLV‐I), type 2, p24 stronger than p 19 (presumptive HTLV‐II), or indeterminate (p 19 and p24 weakly positive or p19 weakly positive in the presence of p21e). Geographic areas could be grouped into 4 patterns. Patterns A (>75% type 1) and B (>75% type 2) were usually observed where the specific type of HTLV or its characteristic diseases had been found. Pattern C (mixed type 1 and 2 pattern) was observed predominantly in intravenous‐drug‐using and other populations in which both virus types have been reported. Pattern D (>10% indeterminate), suggests the presence of non‐specific reactivity, perhaps resulting from exposure to non‐virus‐related antigens or an HTLV‐related virus. HTLV‐I predominates in southern Japan, the South Pacific, parts of West Africa, and in Afro‐populations of the Western hemisphere. HTLV‐II clusters in Native American populations and among intravenous drug users. Pattern‐D areas in Africa and Venezuela might prove to be fertile in the search for new and variant HTLV virus types.