bluuebunny/crossref_metadata_2025 huggingface dataset:
Lymphoproliferative disease in human peripheral‐blood‐mononuclear‐cell‐injected scid mice. II. Role of host and donor factors in tumor generation
Authors: Veronesi Arianna , Coppola Vincenzo , Veronese Maria Luisa , Menin Chiara , Bruni Laura , D'Andrea1 Emma , Mion Marta , Amadori Alberto , Chieco‐Bianchi Luigi
Published: February 2007

Intraperitoneal injection of lymphoid cells from EBV+ donors into SCID mice might provide a useful tool for studying the pathways of B‐cell lymphomagenesis in man. Since previous studies showed that donor T cells greatly favor B‐cell proliferation and tumor generation in this model, we addressed the host and donor factors involved in limiting or promoting lymphoma development. The number of EBV‐infected B‐cell precursors was crucial, since purified B lymphocytes, which alone were unable to generate tumors, underwent expansion and established tumor masses when the animals were inoculated with an EBV‐containing supernatant. Host factors were critical in limiting tumor development; in vivo NK‐cell removal allowed purified B cells to expand and proceed to tumors in the absence of T lymphocytes, whereas potentiation of mouse NK‐cell activity prevented tumor generation in PBMC‐ and LCL‐injected animals. The T‐cell‐derived factors that favor lymphomagenesis could not be identified; IL‐2, IL‐4, IL‐6, and soluble CD23 were not able to promote B‐cell expansion, and treatment of PBMC‐injected mice with the relevant anti‐cytokine anti‐sera did not counteract lymphoma development. These experiments also showed that IL‐6 plays a minor role, if any, in B‐cell lymphoproliferation in this model. Our data indicate that reconstitution of SCID mice with PBMC from EBV+ donors may constitute a useful model for determining the events involved in lymphomagenesis in humans, provided that strict control of all the experimental variables is guaranteed.
Differential effects of protein kinase a sub‐units on chinese‐hamster‐ovary cell cycle and proliferation
Authors: Tortora Giampaolo , Pepe Stefano , Bianco Caterina , Damiano Vincenzo , Ruggiero Angela , Baldassarre Gustavo , Corbo Claudia , Cho‐Chung Yoon Sang , Bianco A. Raffaele , Ciardiello Fortunato
Published: February 2007

It has been shown that a marked increase in the levels of Rlα sub‐units and a decrease in RIIβ sub‐unit levels correlate with neoplastic transformation or with the mitogenic response of normal cells to hormones and growth factors. The selective down‐regulation of RIα and the following induction of RIIβ determine cell‐growth arrest and differentiation of several cancer cells. To directly address the question whether the 2 protein‐kinase‐A(PKA) isoforms play different roles in the control of proliferation and cell‐cycle distribution, we introduced and over‐expressed the different PKA sub‐units in Chinese‐hamster‐ovary (CHO) cells via retroviral‐vector‐mediated gene transfer. Whereas CHO cells treated with RIα anti‐sense oligodeoxynucleotides were growth arrested and accumulated in the G0/G1 phases of the cell cycle, infection of CHO cells with a retroviral vector in order to over‐express RIα determined growth advantages in monolayer conditions and substantially increased their cloning efficiency in soft agar. These events correlated with a sustained percentage of cells in S phase induced by RIα over‐expression in the infected cells. In contrast, CHO cells infected with retroviral vectors over‐expressing either a RIIβ sub‐unit or a Cα catalytic sub‐unit of PKA exhibited growth arrest within a few days of culture and accumulated in the G2‐M phase of the cell cycle. The results of our study demonstrate that the different PKA sub‐units play different and specific roles in the control of cell growth and cell‐cycle distribution.
Brain cancer incidence, mortality and case survival: Observations from two australian cancer registries
Authors: Shugg Dace , Allen Barry J. , Blizzard Leigh , Dwyer Terence , Roder David
Published: February 2007

Data from 2 Australian cancer registries covering a population of 1.7 million people were combined for the purposes of analysing brain cancer incidence, mortality and survival patterns for the time period 1978 through 1992. A total of 1,752 cases of primary brain cancer were registered, representing age‐standardised incidence rates of 6.7 per 100,000 in men and 4.6 in women. Histological confirmation was available for 94% of cases. The incidence rate among persons aged 75 or over was higher during 1986–1992 than during 1978–1985, the rate for men increasing from 16.3 to 26.2 and that for women increasing from 9.7 to 18.0. The largest increases in this age group occurred for cases of glioblastoma multiforme. During the study period, 1,411 brain cancer deaths were notified to the 2 registries at age‐standardised rates of 5.3 in men and 3.4 in women. Mortality rates among persons aged 75 years or older were higher during 1986–1992 than 1978–1985, increasing from 15.7 to 28.4 in men and from 10.1 to 15.3 in women. Only among men aged 15–49 years was a decline in mortality rates observed, from 3.3 to 2.4. Survival analyses indicated that age and histological type were the most powerful prognostic indicators. There was no improvement in 5‐year survival for any of the age groups or histological types. An improvement in 36‐month survival was noted for the 15–49 year age group diagnosed with gliomas other than glioblastoma multiforme.
Immunization with anti‐idiotype monoclonal antibodies bearing the internal image of the renal‐cell carcinoma‐associated antigen G250 induces specific cellular immune responses
Authors: Uemura Hirotsugu , Okajima Eigoro , Debruyne Frans M. J. , Oosterwijk Egbert
Published: February 2007

We have previously isolated and characterized 6 anti‐idiotype antibodies (Ab2s) directed against monoclonal antibody G250 (MAbG250) which reacts with a tumor‐associated antigen (TAA) expressed in a large proportion of human renal‐cell carcinomas (RCC). These 6 Ab2s (NUH31, 51, 71, 82, 91: IgG1, NUH44: IgG2a) showed MAbG250 binding site specificity and induction of anti‐TAA antibody resembling MAbG250 (so‐called Ab l') in rabbits, indicating that they are internal image antibodies. To investigate whether these Ab2s could induce G250‐TAA‐specific cell‐mediated immunity, delayed‐type hypersensitivity (DTH) tests were carried out with G250 antigen‐positive and/or ‐negative cells in the ears of BALB/c mice. Mice primed with Ab2 showed antigen‐specific DTH responses, whereas no significant DTH response was observed with G250‐negative cells. This antigen‐specific DTH could be transferred to naive mice by lymphocytes harvested from Ab2‐sensitized mice. In addition to the classical DTH responses observed 24 and 48 hr after tumor challenge, an early‐phase antigen‐specific hypersensitivity response was seen 2 hr after challenge. This early component of the specific hypersensitivity reaction but not the classical DTH could be transferred to naive mice by serum from Ab2‐sensitized mice, indicating that the early reaction was due to serum factors. These findings demonstrate that all Ab2s induced tumor‐specific cellular immune responses directed against human RCC, and suggest that they may be useful as RCC‐TAA surrogates, i.e., as tumor vaccines for RCC patients.
Effects of type‐I and ‐II interferons on 90K antigen expression in ovarian carcinoma cells
Authors: Marth Christian , Dreps Andreas , Natoli Clara , Zeimet Alain G. , Lang Thomas , Widschwendter Martin , Daxenbichler Günter , Ullrich Axel , Iacobelli Stefano
Published: February 2007

Antigen 90K is produced by several tumor‐cell lines and by patients with cancer. Its function has not yet been clarified, although recent reports suggest that it plays a role in the tumor—host relationship—for example by stimulation of natural killer and lymphokine‐activated killer‐cell activity. Previous studies have indicated that 90K expression may be under the influence of interferon‐α. Here, we provide evidence that both interferon‐α and ‐γ can enhance the secretion of 90K and augment the level of specific mRNA expression in 3 ovarian carcinoma cell lines (OVCAR‐3, HTB‐77 and SKOV‐6). However, interferon‐γ leads to depletion of cellular 90K whereas interferon‐α increases both secreted and cellular 90K levels. In equimolar concentrations, Interferon‐α was always superior to interferon‐γ in augmenting 90K protein or mRNA levels. Combinations of TNF with interferon‐γ were highly synergistic both in reducing cell proliferation and in increasing 90K secretion and mRNA expression. This synergism was seen to a lesser extent with interferon‐α.
Intraperitoneal photodynamic therapy in the rat: Comparison of toxicity profiles for photofrin and mTHPC
Authors: Veenhuizen Ruth B. , Ruevekamp‐Helmers Marjan C. , Helmerhorst Theo J. M. , Kenemans Peter , Mooi Wolter J. , Marijnissen Johannes P. A. , Stewart Fiona A.
Published: February 2007

Toxicity studies for intraperitoneal photodynamic therapy (IPPDT) were performed in Wag/RijA rats, using specially designed light delivery blocks for proper light distribution and light dosimetry. A recently developed photosensitizer mesotetrahydroxyphenylchlorin (mTHPC), excited at 652‐nm wavelength, was compared with Photofrin (630 nm). Toxicity profiles for various sensitizer doses, light fluences and time intervals were investigated. A light fluence of 15 J ‐ cm−2delivered to the entire peritoneum 24 hr after 5 mg Photofrin per kg i.v. induced reversible impairment of intestinal, liver and kidney function. A dose of 0.2 mg mTHPC per kg i.v. followed by 6 J · cm−2at 72 hr appeared to be equitoxic to the intestines; however, functional tests revealed little effect for this mTHPC‐mediated IPPDT regime on liver or kidney. Histology demonstrated focal irreversible damage to the kidneys for both photosensitizers, not reflected in functional impairment. Light doses of 25 to 30 J. cm−2at 24 hr after Photofrin or 8–12 J · cm−2, 72 hr after mTHPC caused lethal toxicity in the first 2 weeks due to intestinal damage. Higher light doses caused a shock syndrome and rhabdomyolysis resulting in death within 20 hr for both photosensitizers. In conclusion, maximum tolerable schedules for whole‐abdomen IPPDT were defined for Photofrin and mTHPC. Both photosensitizers cause similar toxicity profiles depending on drug dose, light fluence and time interval.
Modifications of the anti‐oxidant metabolism during proliferation and differentiation of colon tumor cell lines
Authors: Bravard A. , Beaumatin J. , Dussaulx E. , Lesuffleur T. , Zweibaum A. , Luccioni C.
Published: February 2007

The anti‐oxidant metabolism was studied at different times after sub‐culture in 2 colon cell lines previously characterized for their growth and differentiation properties. The HT29 cell line is mainly composed of proliferative and undifferentiative cells, while the derived 5‐fluorouracil (FUra)‐adapted cells undergo growth‐dependent differentiation, which is complete at post‐confluence. In the 2 cell lines, all the anti‐oxidant parameters studied appeared to be related to proliferation, with increased activity of superoxide dismutase (SOD) 1 and 2, catalase (CAT), glutathione peroxidase (GPX), glutathione reductase (GSR), and glutathione transferase (GST), and decreased glucose‐6‐phosphate dehydrogenase (G6PD) activity and glutathione content, in parallel with slowing down of proliferation. At post‐confluence, these metabolic parameters remained stable, except for GPX activity, which continued to increase, and CAT activity, which decreased. The amounts of SOD1, SOD2 and CAT immunoreactive proteins, estimated by Western blotting, appeared to be correlated to their respective enzymatic activities, SOD1, CAT and GST activity and glutathione content, which remained at similar levels in the 2 cell lines for all times studied, appeared unrelated to the differentiation process. GSR and GPX activity, which was lower in FUraadapted than in parental cells only at post‐confluence, could be considered as markers of differentiated cells. The higher SOD2 and lower G6PD activity observed in FUra‐resistant cell in comparison with parental cells at all times after sub‐culture could be characteristic both of differentiative and of differentiated cells. Interestingly, cytogenetics have previously indicated that deletions of the long arm of chromosome 6, which carry the gene for SOD2, were frequently observed in parental but not in FUra‐adapted cells. These results demonstrate that modifications of the anti‐oxidant metabolism occur in relation with proliferation and differentiation, and suggest a particular role for SOD2 in these cellular processes.
Role of tumor‐derived fibroblasts in the growth of primary cultures of human breast‐cancer cells: Effects of epidermal growth factor and the somatostatin analogue octreotide
Authors: Hofland Leo J. , van der Burg Bart , van Eijck Casper H. J. , Sprij Diana M. , van Koetsveld Peter M. , Lamberts Steven W. J.
Published: February 2007

In the present study we have investigated the role of human breast‐cancer‐derived fibroblasts in the proliferation of primary cultures of epithelial cells derived from the same tumor. For this purpose, a co‐culture system, using Transwell tissue‐culture inserts with microporous membranes was employed. Fibroblasts and epithelial cells were enriched according to differences in their density on Percoll density gradients. The co‐culture system was first established using MCF‐7 breast cancer cells and a human fibroblast line (HF cells). Insulin, 17β‐estradiol, EGF and HF cells all significantly stimulated the growth of MCF‐7 breast cancer cells. The stimulatory effects of insulin, E2 and EGF were additive to the stimulatory effect of HF cells. These data suggest that (unique) factor(s), other than the above‐mentioned growth‐promoting compounds, are responsible for the growth‐promoting effects of fibroblasts. In half of the human breast cancers investigated, tumor‐derived fibroblasts stimulated tumor‐derived epithelial cell proliferation. EGF significantly stimulated epithelial cell proliferation in 4 out of 6 cultures. The stimulatory effects of fibroblasts and EGF were additive or synergistic, and were observed in the additional presence of FCS, again suggesting production of unique factor(s) by the fibroblasts, in one culture the fibroblasts significantly inhibited epithelial tumor‐cell proliferation. Conversely, the epithelial cells significantly stimulated proliferation of fibroblasts in 3 out of 3 cultures. The somatostatin analogue octreotide significantly inhibited epithelial cell proliferation by 46% in one tumor‐cell culture in the absence, but not in the presence, of fibroblasts. In one culture, octreotide significantly inhibited the proliferation of fibroblasts co‐cultured with epithelial cells. © 1995 Wiley‐Liss, Inc.
Nuclear DNA content and chromatin texture in multidrug‐resistant human leukemic cell lines
Authors: Dufer Jean , Millot‐Broguo Christine , Hamed Zohra Oum , Lialtaud‐Roger Francloise , Joly Philippe , Desplaces Andrée , Jardillier Jean‐Claude
Published: February 2007

Nuclear morphological alterations associated with multidrug resistance (MDR) were evaluated by image cytometry in various human leukemic cell sub‐lines: 3 cell lines with P‐gp‐mediated resistance (CEM‐VLB, HL60/Vinc, K562‐Dox), the non‐Pgp‐mediated MDR HL60/AR leukemic cell line with over‐expres‐sion of MRP, and the at‐MDR CEM‐VMI leukemic cell line with alteration of topoisomerase II. All these MDR cell sub‐lines were obtained by drug selection and were compared with their sensitive counterparts and with the hamster LR73‐R cell line obtained by transfection of mouse mdrl cDNA. All MDR cell sub‐lines obtained by drug selection displayed decreased DNA Feulgen stainability as compared with their respective sensitive parental cell line, a phenomenon not observed in the trans‐fected LR73‐R cells. Nuclear texture analysis on G0/G1‐selected cell nuclei revealed 2 types of textural phenotype. The first phenotype was characterized by chromatin decondensation with small but compact chromatin clumps, and was observed in drug‐selected P‐gp‐mediated MDR cells (CEM‐VLB, HL60‐Vinc, K562‐Dox) and in the non‐P‐gp‐mediated MDR HL60/AR cell line. The second phenotype was characterized by a condensed and homogeneous chromatin pattern, and was observed in the at‐MDR CEM‐VMI cell line. LR73‐R cells transfected with mdrl cDNA did not display any significant changes in textural pheno‐type as compared with sensitive LR73 cells, suggesting that P‐gp over‐expression alone cannot account for the cytological modi‐fications observed in MDR cells. These data suggest that multidrug resistance could be associated with specific nuclear morphological changes which appeared to be a consequence of alterations occurring during selection by cytotoxic drugs rather than of P‐gp over‐expression. © 1995 Wiley‐Liss, Inc.
High expression of udp‐n‐acetylglucosamine: β‐d mannoside β‐1, 4‐n‐acetylglucosaminyltransferase III (GnT‐III) in chronic myelogenous leukemia in blast crisis
Authors: Yoshimura Masafumi , Nishikawa Atsushi , Ihara Yoshito , Nishiura Tetsuo , Nakao Hirohisa , Kanayama Yoshio , Matuzawa Yuji , Taniguchi Naoyuki
Published: February 2007

The activity and mRNA expression of UDP‐N‐acetylglucosamine: β‐D mannoside β‐1,4‐N‐acetylglucosaminyl transferase III (GnT‐III: EC 2.4.1.144) were investigated in hematological malignancies. GnT‐III activity was elevated in patients with chronic myelogenous leukemia (CML) in blast crisis and patients with multiple myeloma (MM), as compared to normal healthy subjects and patients with other hematological malignancies including CML in chronic phase. The GnT‐III transcript was the same size in leukemic cells from various hematological diseases and cell lines, while expression of the transcript was not found to correlate significantly with enzyme activity, implying that posttranslational modification might regulate the activity of GnT‐III. Southernblot analysis showed no significant variation in the structure and position of the GnT‐III genome, indicating that the gene is present as a single copy without isoforms. Furthermore, analyses by immunoprecipitation and Western blot revealed that high GnT‐III activity in KU812 cell, a CML cell line, resulted in an increase in E4‐PHA binding to CD45, a major surface glycoprotein of the leukocyte, indicating that more bisecting GlcNAc was added to CD45 catalyzed by elevated GnT‐III.
Inhibition of bispecific monoclonal antibody (bsAb)‐targeted cytolysis by human anti‐mouse antibodies in ovarian carcinoma patients treated with bsAb‐targeted activated T‐lymphocytes
Authors: Lamers Cor H. J. , Gratama Jan W. , Warnaar Sven O. , Stoter Gerrit , Bolhuis Reinder L. H.
Published: February 2007

T lymphocytes of 8 patients with ovarian cancer were targeted to the tumor cells using F(ab')2 fragments of a bispecific monoclonal antibody (bsAb), specific for CD3 (a component of the T lymphocyte receptor for antigen) and for the folate receptor MOv 18 (overexpressed by ovarian carcinoma cells) as part of a phase l/II study. Phase I (days 0 to 3) consisted of increasing intraperitoneal (i.p.) numbers (106−109) of bsAbtargeted T lymphocytes plus lowdose interleukin‐2 (IL‐2). Phase II (days 6 to 13, and 27 to 33) consisted of daily i.p. infusions of 109 targeted T lymphocytes, 2 mg soluble bsAb, and lowdose IL‐2. Using enzymelinked immunosorbent assays (ELISA), human antimouse antibodies (HAMA) were detected in all patients: in the serum from day 13 onwards and in the peritoneal fluid from day 20 onwards. A significant proportion of the HAMA appeared to be directed against the idiotypes of the bsAb specific for CD3 and MOv18, as suggested by (I) the clearly higher ELISA titers against OC/TR bsAb as compared to those against a monoclonal antibody (MAb) with unrelated specificity, and (2) failure to abrogate the capacity of peritoneal fluid containing HAMA to block the binding of OC/TR bsAb to MOv18+ or CD3+ cells by absorption of human antimouse IgG‐framework antibodies in peritoneal fluid to immobilized mouse IgG. The OC/TR‐targeted cytolysis of the MOv18+ ovarian carcinoma cell line lgrov‐1 by autologous T lymphocytes was inhibited by peritoneal fluid samples containing relatively high HAMA titers. Such inhibitory activity was never detected at the start of phase II, but coincided with the last series of i.p. infusions of targeted T lymphocytes in 2 patients.
A low incidence of p53 mutations in pre‐malignant lesions of the oral cavity from non‐tobacco users
Authors: Lazarus P. , Garewal H. S. , Sciuuba J. , Zwiebel N. , Calcagnotto A. , Fair A. , Schaefer S. , Richie J. P.
Published: February 2007

To determine the incidence of pS3 mutations in pre‐malignant lesions of the oral cavity from individuals without prior history of tobacco use, we have analyzed the conserved regions of the pS3 gene (exons 5‐9) in archival oral cavity lesion specimens obtained from patients with varied tobacco use histories, by polymerase chain reaction/single strand conformational polymorphism (PCR/SSCP) and DNA sequencing analysis. Twenty‐six lesions were analyzed from 14 patients, with multiple lesions obtained from 8 patients. Six of these patients used tobacco, (3 being cigarette smokers, 1 ex‐cigarette smoker, 1 moderate cigar smoker and 1 snuff chewer). The remaining 8 patients had no prior history of tobacco use. Thirteen of the pre‐malignant lesions exhibited severe dysplasia, 9 exhibited moderate dysplasia and 4 exhibited mild dysplasia. Four of the 26 lesions exhibited pS3 mutations, each being from a tobacco user. None of the 13 lesions from never‐tobacco users exhibited pS3 mutations. There was a significantly higher p53 mutation incidence in pre‐malignant lesions from tobacco users (including ex‐smokers) than in non‐tobacco users as well as in cigarette smokers plus snuff chewers than in non‐tobacco users. Two of the mutations were observed in lesions exhibiting severe dysplasia: I in a lesion exhibiting moderate dysplasia and I in a lesion exhibiting mild dysplasia. These data suggest that pS3 mutation may be a very early event in oral cavity tumor progression and demonstrate that pre‐malignant lesions obtained from non‐tobacco users do not exhibit pS3 mutations.
Risk of subsequent malignant neoplasms among 470,000 cancer patients in Finland, 1953‐1991
Authors: Sankila Risto , Pukkala Eero , Teppo Lyly
Published: February 2007

Because of increasing numbers of incident cancers and improving survival rates, more and more cancer patients are at risk of subsequent primary malignant neoplasms. The potential carcinogenicity of cancer therapies raises further concern regarding this unwanted long‐term side effect. The objective of this cancer‐registry‐based study was to assess the trends in the risk of subsequent primary malignancies among cancer patients on a nationwide public‐health level. The 470,000 cancer patients registered between 1953 and 1991 in Finland were followed up for subsequent primary neoplasms, and 19,800 subsequent malignant neoplasms were observed, yielding an overall standardized incidence ratio (SIR) of 1.00 for men and 1.25 for women. Substantial variation was observed in the SIR by the site of subsequent malignancy. The adjusted relative risk derived from multiple regression analyses showed only a slight increase by follow‐up interval. Overall, the excess relative risk of subsequent malignancies was low, since the majority of cancer patients are old and at low relative risk. Patients under the age of 50 years, however, were at 70% increased risk, and the overall relative risk increased by 50% from the 1950s to the 1980s. Subsequent neoplasms among cancer patients do not pose a major public‐health problem, but the possibility of subsequent cancers must be borne in mind in decisions concerning therapy and in clinical follow‐up.
Analysis of the NF2 tumor‐suppressor gene and of chromosome 22 deletions in gliomas
Authors: Hoano‐Xuan Khě , Meree Philippe , Vega Felipe , Hugot Jean‐Pierre , Cornu Philippe , Delattre Jean‐Yves , Poisson Michel , Thomas Gilles , Delattre Olivier
Published: February 2007

Recurrent deletions of chromosome fragments observed in neoplasms are thought to participate in tumor development through the inactivation of tumor‐suppressor genes. In gliomas, the most frequent deletions involve chromosome arms 9p, 10q, 17p, 19q and 22q. We have analysed deletions of chromosome 22 in gliomas by studying loss of heterozygosity (LOH) at 8 microsatellite loci. LOH for this chromosome fragment was observed in 17/70 (24%) cases, most of them encompassing the region which encodes the gene altered in neurofibromatosis 2 (NF2), an inherited disease which predisposes to tumors of the nervous system. To investigate the possible involvement of the NF2 tumor‐suppressor gene in the tumorigenesis of gliomas, we searched for alterations in its genomic structure and in its mature transcript. Northern‐blot and reverse transcriptase‐PCR experiments showed that the NF2 transcript is expressed and does not demonstrate obvious structural alterations. Moreover, analysis, at the genomic level, of the 16 coding exons of the NF2 gene by denaturing gradient gel electrophoresis failed to detect any somatically acquired point mutations. Altogether, these data strongly suggest that, although gliomas demonstrate recurrent chromosome 22 deletions most frequently encompassing the NF2 region, the NF2 gene is not altered in these tumors.
Proteolytic enzymes at various stages of oncogenic transformation of rat fibroblasts. I. Aspartyl and cysteine proteinases
Authors: Solovyeva N. I. , Balayevskaya T. O. , Dilakyan E. A. , Zakamaldina‐Zama T. A. , Pozdnev V. F. , Topol L. Z. , Kisseljov F. L.
Published: February 2007

Aspartyl and cysteine proteinases at distinct stages of carcinogenesis were analyzed in rat embryo fibroblasts, sequentially immortalized and transformed by 2 different genes: the early region of simian adenovirus SA7 and c‐Ha‐ros oncogene. The dynamics of expression and distribution of proteinases throughout the transformation process were examined. It was shown that in immortalized and transformed cells the activities of the aspartyl and cysteine proteinases were expressed to a variable degree and that the expression was dependent on cell‐propagation time in vitro. The increase in activity both of cathepsin‐D‐like aspartyl proteinase and of cathepsin‐L‐ and ‐B‐like cysteine proteinases in cell lysates was correlated with the stages of fibroblast transformation (immortalization and tumorigenic transformation). In all cell types the major part of cysteine proteinases was localized inside the cell, while the cathepsin‐D‐like proteinase was apparently predominant among secreted proteinases. The cathepsin‐L‐like proteinase accounts for the major part of the cysteine‐proteinase activity as measured by Z‐Phe‐Arg‐MCA hydrolysis. We suggest that considerable portions of the cathepsin‐D‐ and ‐L‐like proteinases in all cell lines studied are secreted as a complex with inhibitor(s) and that inhibitor expression plays an important role in regulating the activity of cathepsin‐D‐like proteinase at different stages of transformation. Cathepsin‐L‐like proteinase is probably secreted in the precursor form.
Effect of VK framework‐1 glycosylation on the binding affinity of lymphoma‐specific murine and chimeric LL2 antibodies and its potential use as a novel conjugation site
Authors: Leung Shui‐On , Dion Arnold S. , Pellegrini Matthew C. , Losman Michele J. , Grebenau Ruth C. , Goldenberg David M. , Hansen Hans J.
Published: February 2007

A potential asparagine (Asn)‐linked glycosylation site was identified in the VK FRI sequence of an anti‐B lymphoma monoclonal antibody (MAb), LL2. SDS‐PAGE analysis and endo‐F treatment of both murine and chimeric LL2 antibodies indicated that this site was glycosylated; however, no differences in the binding affinity to Raji cells were observed between the native murine LL2 and the endo‐F‐deglycosylated murine LL2 antibodies. Elimination of the glycosylation site from the chimeric LL2 antibody was accomplished by an Asn to Gin mutation in the tri‐acceptor site found in the light chain. The resultant aglycosylated chimeric LL2 exhibited a similar Raji cell binding affinity to that of the glycosylated form. The results are in agreement with computer modeling studies which suggested the lack of interactions between the oligosaccharide moiety and the CDRs. The finding is interesting because it enables a wider choice of human framework sequences, which in most cases do not have a corresponding glycosylation site, for the humaniza‐tion of the LL2 VK domain, as well as a greater latitude of host expression systems. Most importantly, the LL2 VK carbohydrate moiety might be used as a novel conjugation site for drugs and radionuclides without compromising the immunoreactivity of the antibody.
Shedding of the soluble form of CD30 from the Hodgkin‐analogous cell line L540 is strongly inhibited by a new CD30‐specific antibody (Ki‐4)
Authors: Horn‐Lohrens Ottmar , Tiemann Markus , Lange Hans , Kobarg Jörg , Hafner Martin , Hansen Hinrich , Sterry Wolfram , Parwaresch Reza M. , Lemke Hilmar
Published: February 2007

The CD30‐activation marker was detected as the Hodgkin‐associated Ki‐I antigen and is regarded as a target for the treatment of Hodgkin patients with immunotoxins. The CD30 is released from tumor cells and this soluble CD30 (sCD30) is an indicator of the disease activity. Since the shedding of sCD30 may be influenced by antibodies, we produced 6 new CD30‐specific antibodies (Ki‐2 to Ki‐7) for the purpose of finding antibodies that might inhibit the formation of sCD30. Ki‐2 to Ki‐7 and the other anti‐CD30 antibodies Ki‐I, Ber‐H2, HeFi‐l, M44, M67, HRS‐I, HRS‐4 and CIO were employed for epitope mapping. The binding of a particular radio‐labeled anti‐CD30 antibody to Hodgkin's‐disease‐derived L540 cells was completed by addition of the various non‐labeled anti‐CD30 antibodies. Three non‐overlapping regions, expressing different antigen‐specific determinants, could be defined on the extracellular part of the CO30 molecule. Cluster A of determinants was recognized by Ki‐2, Ki‐4, Ki‐6 and Ki‐7, Ber‐H2, HRS‐I and HRS‐4, while cluster B was detected by Ki‐I, Ki‐S and M67. Cluster C, which probably contains the binding site for the CD30 ligand, was defined by Ki‐3, M44, HeFi‐l and CIO. Co‐culture experiments of LS40 cells with the various antibodies followed by the isolation of sCD30 from culture supernatant fluids revealed that the release of sCD30 was most strongly increased by Ki‐I and weakly enhanced by Ki‐2, Ki‐3, Ki‐5 and HeFi‐l, whereas it was almost completely inhibited by Ki‐4 and to a slightly lesser extent by Ber‐H2.
Expression of a B‐cell marker, CD24, on nasopharyngeal carcinoma cells
Authors: Karran Lorainc , Jones Michael , Morley Gary , Van Noorden Susan , Smith Paul , Lampert Irvin , Griffin Beverly E.
Published: February 2007

Random sequencing of clones from a λgt1O cDNA library, made from mRNA expressed in an Epstein‐Barr virus (EBV)‐associated nasopharyngeal carcinoma (NPC) has revealed the gene transcript of human CD24. The CD24 antigen, a glyco‐sylphosphatidylinositol‐anchored cell surface molecule, has been identified as a B‐cell marker that is lost during cell maturation. We show here that it is expressed on 3 NPC xenografts, previously defined as consisting of poorly differentiated epithelial cells, and on an NPC biopsy. In the case of the former, the level of expression of CD24 corresponds to the EBV load. A B‐lymphoblastoid cell line carrying the same EBV genome as one of the tumours, C15, and an EBV‐negative Burkitt's lymphoma cell line do not display the antigen, but epithelial‐like cells of a laryngeal tumour cell line (Hep2) do express it. Our data suggest that CD24 may be a marker of cell differentiation not only for B cells but also for epithelial cells and may have an indirect association with EBV gene expression.
Vaccination of rabbits with recombinant vaccinia virus carrying the envelope gene of human T‐cell lymphotropic virus type I
Authors: Hakoda E. , Machida H. , Tanaka Y. , Morishita N. , Sawada T. , Shida H. , Hoshino H. , Miyoshi I.
Published: February 2007

Two groups of 3 rabbits each were immunized with either recombinant vaccinia virus, WR‐SFB5env, carrying the human T‐cell lymphotropic virus type I (HTLV‐1) env gene at the site of the hemagglutinin gene of the WR strain, or control vaccinia virus, HA‐WR, lacking the functional hemagglutinin gene. All 6 rabbits responded with anti‐vaccinia virus antibodies. WR‐SFB5env elicited anti‐HTLV‐1 env antibodies but no vesicular stomatitis virus (HTLV‐1) pseudotype neutralizing antibodies in all 3 rabbits. After 10 weeks, the animals were challenged by transfusion of blood from an HTLV‐1‐infected rabbit. Two of the 3 vaccinated rabbits and all 3 control rabbits became infected with HTLV‐1, as indicated by seroconversion and detection of HTLV‐1 proviral sequences by polymerase chain reaction. The rabbit that had been protected from initial challenge became infected with HTLV‐1 upon rechallenge 12 weeks after the first challenge. In view of the proven prophylactic effect of passive immunization against HTLV‐1, our vaccine trial failed because WR‐SFB5env was incapable of inducing neutralizing antibodies against HTLV‐1 in the immunized animals. It remains to be studied whether cell‐mediated immunity such as antibody‐dependent cellular cytotoxicity was involved in the temporary protection of I vaccinated rabbit.
Risk of ocular melanoma in relation to cutaneous and IRIS naevi
Authors: Bataille Veronique , Sasieni Peter , Cuzick Jack , Hungerford John L. , Swerdlow Anthony , Bishop Julia A. Newton
Published: February 2007

A case‐control study was set up to assess the risk of eye melanoma in relation to the number and type of cutaneous melanocytic naevi and pigmented lesions of the iris. Cases comprised 211 unselected ocular melanoma patients attending the Ocular Oncology Clinic at Moorfields Eye Hospital, London, during November 1990 to October 1991 and diagnosed after August 1986. Hospital and general practice controls (416) were recruited in the North East Thames Region of the UK. Cutaneous naevi greater than or equal to 2 mm in diameter were counted on the skin. Clinically atypical and congenital naevi were recorded separately. Pigmented lesions of the iris were counted. The relative risk for ocular melanoma increased with numbers of atypical naevi and numbers of common naevi. Ten percent of cases but 3% of controls had at least 100 naevi of 2 mm or greater diameter. Seven percent of cases and 0.4% of controls had 4 or more atypical naevi. Pigmented lesions of the iris were significantly more common in cases than controls. Nine percent of cases had the Atypical Mole syndrome (AMS) phenotype compared with 1% of controls. Six cases had concurrent cutaneous melanoma primaries. We conclude that atypical and iris naevi are important risk factors for eye melanoma and that patients with eye melanoma are at increased risk of cutaneous melanoma. Dermatological examination for the AMS phenotype and cutaneous melanoma should be recommended in eye melanoma patients with large numbers of pigmented lesions of the skin or family history of melanoma.
A case‐control study of hepatitis B and C virus infections in the etiology of hepatocellular carcinoma
Authors: Hadziyannis Stephanos , Tabor Edward , Kaklamani Evangclia , Tzonou Anastasia , Stuver Sherri , Tassopoulos Nicolaos , Mueller Nancy , Trichopoulos Dimitrios
Published: February 2007

During a 16‐month period in 1991‐1992, blood samples and questionnaire data were obtained from 65 incident cases of hepatocellular carcinoma (HCC) as well as from 2 control groups of hospitalized patients matched on gender and age, which included 65 metastatic liver cancer (MLC) patients and 65 patients hospitalized for eye, ear, nose or throat conditions. Coded sera were tested for hepatitis B surface antigen (HBsAg), antibody to hepatitis B core antigen, antibody to HBsAg and antibody to hepatitis C virus (anti‐HCV) by enzyme immunoassay. The odds ratios (with 95% confidence intervals) in logistic regression modeling comparing the HCC cases to the combined control series were 18.8 (8.2–43.2) for the presence of HBsAg and 7.7 (1.7–35.1) for anti‐HCV. In the present hospital‐based case‐control study anti‐HCV testing was conducted on recently collected sera, using a second‐generation enzyme immunoassay with confirmation by immunoblot assay. Comparisons with previous work in a similar population demonstrated that, when second‐generation anti‐HCV assays are applied to sera stored for 7–15 years, confirmatory assays or a higher diagnostic cut‐off point may be necessary to ensure that the testing is specific.
Expression of transforming growth factor alpha and its receptor in human neuroendocrine tumours
Authors: Nilsson Ola , Wängberg Bo , Kölby Lars , Schultz Gregory S. , Ahlman Håkan
Published: February 2007

Transforming growth‐factor‐α (TGF‐α) is a 50‐amino‐acid polypeptide that binds to the epidermal growth factor (EGF) receptor and stimulates cell growth. It has been suggested that enhanced production of TGF‐α and EGF receptors by tumour cells promote tumour‐cell growth by autocrine mechanisms. In the present study we have investigated the expression of TGF‐α and EGF receptors in human neuroendocrine tumours, including midgut carcinoid tumours, phaeochromocytomas and medullary thyroid carcinomas. TGF‐α expression was demonstrated in biopsies of all tumours examined (n = 30) and EGF receptors in a majority of tumours by Northern analysis and/or immunocy‐tochemistry. Expression of TGF‐α and EGF receptors was also demonstrated in primary cultures of tumour cells. Carcinoid tumours and phaeochromocytomas in culture secreted detectable amounts of TGF‐α into the culture medium (400–700 pM). The amount of secreted TGF‐α could be suppressed by octreotide treatment in individual tumours. Administration of exogenous TGF‐α stimulated carcinoid tumour growth in vitro as determined by the DNA contents of cell cultures. The growth‐stimulatory effect of TGF‐α could be partially blocked by the use of neutralizing anti‐EGF receptor monoclonal antibodies (MAbs). In conclusion, several human neuroendocrine tumours express both TGF‐α and EGF receptors in in vivo and in vitro, suggesting that TGF‐α may regulate tumour‐cell growth by autocrine mechanisms.
Characterization of selected strongly and weakly invasive sublines of a primary human melanoma cell line and isolation of subtractive cDNA clones
Authors: Jacob Karin , Bosserhoff Anja K. , Wach Franz , Knüchel Ruth , Klein Eberhard C. , Hein Rüdiger , Buettner Rcinhard
Published: February 2007

Invasion of basement membranes is a key step in systemic spread of tumour cells. To analyze genetic mechanisms involved in this process, we have selected strongly and weakly invasive sublines with stable phenotypes from a primary human melanoma cell line by repeated passage through a reconstituted basement membrane in vitro. The sublines differed approximately 5‐fold in their invasive potential. Invasiveness correlated with better attachment and overexpression of the integrin αv/β3 (vitronectin/laminin‐receptor). Treatment with retinoic acid inhibited proliferation in both sublines and invasion in the weakly invasive cells but stimulated invasion in the strongly invasive subline. Northern‐blot analyses revealed equal levels of mRNA expression regarding collagenase type‐IV and retinoic‐acid receptors but enhanced expression of TIMP‐2 mRNA in weakly invasive cells. The 2 sublines differed significantly in their respective DNA ploidy when compared to the wild‐type Mel Im cell line, suggesting that they represent heterogeneous clones present in the primary tumour. We have started to exploit this in vitro system for tumour heterogeneity to clone genes involved in invasion. By a subtractive cDNA cloning strategy, 12 partial cDNA clones were obtained that are specifically overexpressed in the strongly or weakly invasive subline. These results illustrate that stable genetic alterations lead to heterogeneous subpopulations within primary melanomas which differ in their ability to invade basement membranes and interact with components of the extracellular matrix.
Treatment of Philadelphia‐chromosome‐positive human leukemia in scid mouse model with herbimycin A, bcr‐abl tyrosine kinase activity inhibitor
Authors: Honma Yoshio , Matsuo Yoshinobu , Hayashi Yasuhide , Omura Satoshi
Published: February 2007

The molecular basis of the Philadelphia chromosome (Ph) is a structurally altered c‐abl (bcr‐abl) gene which encodes an abnormally large protein with protein tyrosine kinase activity. Herbimycin A, which effectively reduced intracellular phosphorylation by bcr‐abl tyrosine kinase, preferentially inhibited the growth of Ph‐positive leukemia cell lines. Injection of Ph‐positive and ‐negative leukemia cell lines into mice with severe combined immunodeficiency (SCID) resulted in the death of all mice due to leukemia, although the severity of illness varied according to the cell lines used. Administration of herbimycin A significantly enhanced the survival of mice inoculated with the Ph'‐positive leukemia cell lines tested but barely affected the survival of mice inoculated with the Ph‐negative leukemia cell lines tested. These results suggest that herbimycin A and related compounds may be useful for the treatment of Ph‐positive leukemia. The disease that developed using the Ph‐positive leukemia cell line NALM‐20 resembled human Ph‐positive acute lymphoid leukemia. There was an inverse relationship between the survival time of mice and the number of cells inoculated. The SCID mouse‐NALM‐20 human leukemia chimera would be a good experimental model for screening tyrosine kinase inhibitors as therapeutic agents against Ph‐positive leukemia.
Somatostatin analog RC‐160 inhibits growth of CFPAC‐1 human pancreatic cancer cells in vitro and intracellular production of cyclic adenosine monophosphate
Authors: Qin Yunfcng , Ertl Tibor , Groot Kate , Horvath Judit , Cai Ren‐Zhi , Schally Andrew V.
Published: February 2007

The effect of somatostatin analog RC‐160 on the growth of CFPAC‐I human pancreatic cancer cells in vitro was investigated. RC‐160 effectively inhibited the proliferation of CFPAC‐I cells in culture, inducing a time‐ and dose‐dependent decrease in the number of treated cells. A significant suppression of cell growth was observed after 48 and 72 hr of the exposure to (I μM) RC‐160, the cell number being decreased by 38% and 46%, respectively. RC‐160 was more potent than SS‐14 or SMS201–995 in inhibiting the growth of CFPAC‐I cells, and after 48‐hr treatment the cell number decreased by 49% for RC‐160 compared with 12% for SS‐14 and 27% for SMS201‐995. Binding experiments demonstrated that specific receptors for somatostatin were present on CFPAC‐I cells. SS‐14 showed a high binding affinity for [125I]‐Tyr11‐SS‐14 receptors on CFPAC‐I cells. Scatchard analysis indicated the presence of 2 classes of somatostatin binding sites on the cells, one with high binding affinity and low capacity and the other with low binding affinity and high capacity. RC‐160 could bind to somatostatin receptors on these cells with an affinity similar to SS‐14 but significantly higher than that of SMS201–995. Radioimmunoassay of intracellular cAMP showed that RC‐160 could powerfully inhibit forskolin‐stimulated cAMP production in CFPAC‐I cells. Addition of forskolin to the cultures increased cAMP concentrations in the cellular lysate of treated cells. RC‐160 attenuated or nullified in a dose‐dependent manner the cAMP production stimulated by forskolin. Our observations indicate that somatostatin analog RC‐160 inhibits the proliferation of CFPAC‐I human pancreatic cancer cells in vitro and that this effect may involve the intracellular cAMP pathway.
E‐selectin expression induced by pancreas‐carcinoma‐derived interleukin‐1α results in enhanced adhesion of pancreas‐carcinoma cells to endothelial cells
Authors: Kaji Mitsuhito , Ishikura Hiroshi , Kishimoto Takashi , Omi Makoto , Ishizu Akihiro , Kimura Chisa , Takahashi Toshiyuki , Kato Hiroyuki , Yoshiki Takashi
Published: February 2007

Cellular adhesion of sialyl‐Lewis‐a(SLea)‐positive pancreas carcinoma to endothelial cells (EC) is augmented by activation of EC via up‐regulated E‐selectin expression on EC. Co‐cultivation of pancreas‐carcinoma cells, PCI‐24, with human umbilical‐vein endothelial cells (HUVEC) for 5 hr at the PCI‐to‐HUVEC ratio of 1:10 induced E‐selectin expression on the endothelial‐cell surface, augmenting SLea‐positive pancreas‐carcinoma cell attachment with HUVEC. Culture supernatants of 6 tested pancreas‐carcinoma cell lines contained soluble, E‐selectin‐inducing factor(s). The E‐selectin‐inducing effect by the supernatants was blocked by the protein‐kinase‐C inhibitor, H7. Antibodies against SLea and E‐selectin but not SLex or ICAM‐I blocked the increased pancreas‐carcinoma‐to‐endothelial attachment. Paraformaldehyde(PFA)‐fixed PCI‐24 cells also induced E‐selectin on vascular endothelial cells upon direct contact with endothelial cells, indicating the presence of a membrane‐bound form. The 6 pancreas‐carcinoma lines all produced IL‐1α mRNA and protein but not IL‐1β or TNF‐α protein and/or mRNA Absorption of IL‐lα from the supernatants by IL‐lα‐specific antibody almost completely abolished E‐selectin‐inducing activity. Anti‐IL‐lα antibody also abolished the E‐selectin‐inducing activity of PFA‐fixed PCI. IL‐1α production by PCI cells was up‐regulated by TNF‐α. These observations suggest that substance(s) produced by pancreas‐carcinoma cells, in this case, IL‐1α, may contribute to pancreas‐carcinoma‐cell colonization in non‐inflamed, distant locations in vivo, by activating vascular endothelial cells.
A mutational hot spot in the p53 gene is associated with hepatoblastomas
Authors: Oda Hideaki , Nakatsuru Yoko , Imai Yasuo , Sugimura Haruhiko , Ishikawa Takatoshi
Published: February 2007

Hepatoblastomas generally appear in children aged 2 or 3 years old and arise from apparently normal, noncirrhotic liver. To elucidate any possible role of p53 mutations in their genesis, we amplified and sequenced exons 5 to 8 of the p53 gene in 10 cases of hepatoblastoma. Somatic mutations were detected in 9 cases, in eight of which a common point mutation at the firstbase position of codon 157 was found, resulting in an aminoacid substitution of phenylalanine for valine. Two missense mutations in codon 244, and one each in codons 273 and 279, were also found, with 3 hepatoblastomas having double missense mutations. Out of the total of 12 mutations, 11 were G‐to‐T transversions. One was a G‐to‐A transition and guanines were always present on the transcribed strand. Furthermore, p53 overexpression was immunohistochemically observed in 7 out of 9 cases with p53 gene mutations, although the staining pattern was focal and heterogeneous. The findings suggest that particular environmental mutagens may be involved in mutagenesis of the p53 gene in some cases of hepatoblastomas and that p53 mutations at a specific site may play an important role in the genesis of this disease. © 1995 Wiley‐Liss, Inc.
Generation of CD4+ cytotoxic T lymphocytes stimulated by immobilized anti‐CD3 monoclonal antibody and interleukin‐2 in cancer patients
Authors: Tani Masaji , Tammura Hiroshi , Yamaue Hiroki , Mizobata Shizuma , Iwahashi Makoto , Tsunoda Takuya , Noguchi Kohci , Tamai Mikiko , Hotta Tsukasa , Terasawa Hiroshi , Arii Kazuo
Published: February 2007

The proliferation of autologous tumor‐reactive cytotoxic T lymphocytes (CTL), induced by autologous mixed lymphocyte tumor‐cell culture, was remarkably enhanced by activation with immobilized anti‐CD3 monoclonal antibody (MAb) and interleukin‐2 (IL‐2), as compared with IL‐2 alone. The activated CTL exhibited high cytotoxicity against autologous tumor cells. Cytotoxicity against autologous tumor cells was inhibited by anti‐HLA‐DR MAb. In negative selection with immunomagnetic beads, cytotoxicity against autologous tumor cells was inhibited by the elimination of CD4+ cells. The major cell‐surface antigens of the activated CTL were CD3+, CD4+, CD25+, CD45RO+and CD45RA‐, suggesting helper T cells, and the activated CTL produced IL‐2. It is concluded that the CTL activated by immobilized anti‐CD3 MAb and IL‐2 were CD4 cells that had both killer and helper functions. Our findings indicate that adoptive immunotherapy using these activated CTL would be effective in cancer patients. © 1995 Wiley‐Liss, Inc.