bluuebunny/crossref_metadata_2025 huggingface dataset:
Malignant melanoma: Cross‐reacting (common) tumor rejection antigens
Authors: Hearing Vincent J. , Vieira Wilfred D. , Law Lloyd W.
Published: February 2007

The expression of tumor‐associated transplantation antigens (TATA) by 3 different murine melanomas was examined. A comparison was made between different modes of inducing tumor‐rejection activity, including immunization with irradiated cells from tissue culture lines, with irradiated cells from solid tumor lines, and with viable cells growing in footpads (followed by amputation). Melanoma cell lines examined included the spontaneous B 16 melanoma, the ultra violet‐light‐induced K 1735 melanoma, and the dimethyl benzanthracene‐induced JB/RH melanoma. The data presented demonstrate that not only do all 3 melanoma lines studied express cell surface antigens sufficient to elicit immune response which result in tumor rejection activity, but that these antigens show cross reactivity among the 3 melanoma lines studied. The specificity of the TATA appears to be restricted to the melanomas, since crossreactivity was not observed with 2 different fibrosarcoma cell lines, or with 2 sarcoma cell lines. In addition, it was found that both the JB/RH and K 1735 melanoma cells release (or shed) cell surface antigens which can elicit tumor rejection activity, and that these antigens can be extracted with aqueous butanol, as has been demonstrated with B 16 melanoma.
Procoagulant activity of mouse and human cultured cells following various types of transformation
Authors: Curatolo Laura , Alessio Grazia , Passerini Carlo Gambacorti , Casali Bruno , Morasca Luciano , Semeraro Nicola , Donati Maria Benedetta
Published: February 2007

The presence of fibrin deposits in the micro environment of tumor cells has been reported repeatedly and considered to play an important role in tumor biology. Among the mechanisms by which fibrin may be deposited in tumors, procoagulant activities (PCA) of different types have been described in cancer cells. The present study was aimed at establishing whether the nature of cellular PCA was a characteristic associated with malignant transformation. PCA of normal and transformed cells was investigated on pairs of murine and human origin. The transformed counterparts were obtained after treatment with low‐dose radiation, chemical carcinogen, viral infection or after in vitro spontaneous immortalization. Both before and after any type of transformation cell PCA was of the tissue thromboplastin type, identified on the basis of biological criteria: requirement of factor VII for its expression and lack of inhibition by the serine protease inhibitor diisopropylfluorophosphate (DFP). Transformed cells of murine origin showed significantly lower activity than their normal counterparts, whereas all the transformed human cell lines expressed significantly higher activity than normal. An inverse correlation between the levels of PCA and the cell density in culture was observed in all but one of the lines tested. These findings suggest that the factor X activating property described in some tumors or in transformed cells cannot be considered as a general marker of transformation.
A dual expression of cytokeratin and neurofilaments in bronchial carcinoid cells
Authors: Lehto Veli‐Pekka , Miettinen Markku , Virtanen Ismo
Published: February 2007

Intermediate filaments (IF) are ubiquitous cytoplasmic structures which, by virtue of their cell‐ and tissue‐type‐specific characteristics, are widely used as markers of tissue derivation and as differential diagnostic aids in surgical pathology. In contradistinction to other IFs, vimentin filaments, characteristic of mesenchymal cells, may be coexpressed with other cell‐type specific IFs—cytokeratin filaments, desmin filaments, glial filaments and neurofilaments—in some tumor cells, embryonic cells, and cells in vitro. In this study we describe a novel type of IF coexpression which does not involve vimentin‐filaments, viz. the presence of both cytokeratin filaments and neurofilaments in human bronchial carcinoid tumor cells.
Distinctions between endemic and sporadic forms of epstein‐barr virus‐positive burkitt's lymphoma
Authors: Rowe M. , Rooney C. M. , Rickinson A. B. , Lenoir G. M. , Rupani H. , Moss D. J. , Stein H. , Epstein M. A.
Published: February 2007

Tumour cell lines were established in vitro from 16 cases of Epstein‐Barr (EB) virus genome‐positive Burkitt's lymphoma (BL), 7 of “endemic” origin (i.e. from holoendemic malarial areas of Africa and of New Guinea) and 9 of “sporadic” origin (i.e. from outside such high‐incidence areas). All the BL cell lines thus established were monoclonal by immunoglobulin iso‐type expression and displayed a characteristic chromosomal translocation, t(8:14) or t(8:22), confirming their malignant origin. Clear differences observed between the individual BL cell lines appeared to be related to their endemic or sporadic status. All 7 endemic cell lines began growth as a carpet of single cells, often with small, loose cumps appearing in later passage. Whilst 3 lines of sporadic origin displayed a similar pattern to the above, the majority of sporadic lines grew as large, tight clumps of cells from the first passage onwards. These differences in growth pattern were reflected by differences in cell surface phenotype, as defined in indirect immunofluorescence tests using a panel of monoclonal antibodies (MAbs) specific for B‐lineage‐associated antigens. BL cell lines could be classified into 3 separate groups on the basis of their reactivity with 6 particular antibodies (MHM6, AC2, Ki‐1, Ki‐24, J5 and 38.13). All 7 endemic BL cell lines and 2 of the 3 sporadic BL cell lines which began growth as single cells showed a group‐I cell‐surface phenotype (MHM6, AC2, Ki‐1, Ki‐24 negative; J5, 38.13 positive) in early passage. In contrast, all 6 sporadic BL cell lines which began growth in large clumps displayed a distinct group‐11 phenotype (MHM6, AC2, Ki‐1 positive/negative; Ki‐24, J5, 38.13 positive); in later passage most of these sporadic lines progressed to a group‐III phenotype (MHM6, AC2, Ki‐1, Ki‐24 positive; J5, 38.13 negative) without loss of those immunoglobulin and chromosomal markers identifying the cells' malignant origin. These clear differences between endemic BL cell lines on the one hand and the majority of sporadic BL cell lines on the other suggest that endemic BL arises from a more restricted range of progenitor B cells than does the sporadic form of the disease.
Papillomavirus infection of the lower genital tract: Detection of viral DNA in gynecological swabs
Authors: Schneider Achim , Kraus Herbert , Schuhmann Roland , Gissmann Lutz
Published: February 2007

A total of 311 smears from the lower genital tract were examined by the filter in situ hybridization method to identify human papillomavirus (HPV) DNA.

Of these 311 smears, 229 came from clinically and cytologically negative patients and served as a control group. In this group HPV‐DNA was detected in 5 cases (2.2%).

Of 82 cytologically positive cases (25 confirmed by histology) 56 (68%) contained HPV‐DNA. A high prevalence of HPV 6/11 and absence of HPV 16/18 was found in cases with cytological signs of permissive HPV infection. In mild and moderate dysplasia all viruses occurred at almost the same frequency. In severe dysplasia/carcinoma in situ HPV 16/18 was found 5 times more frequently than HPV 6/11. HPV 16/18 was identified in all 4 invasive cancer cases.

Cervical irrigation of colposcopically suspect areas was performed in 15 cytologically and HPV‐DNA positive cases using the hydrodynamic filtration method. In 12 cases only the cells obtained from the colposcopically positive areas contained HPV‐DNA.

The sensitivity and reproducibility of the filter in situ hybridization was shown by: (1) comparing the results obtained by HPV‐DNA hybridization using Southern blot analysis of tumor biopsies; (2) analysing the correlation of cytologic diagnosis and presence of HPV‐DNA in follow‐up examinations, and (3) diagnosing presence or absence of HPV‐DNA in parallel filters from the same patients.
Detection of nk activity and antibody‐dependent cellular cytotoxicity of lymphocytes by human tumor clonogenic assay‐its correlation with the 51cr‐release assay
Authors: Nomori Hiroaki , Saljo Nagahiro , Fujita Jiro , Hyoi Megumi , Sasaki Yasutsuna , Shimizu Eiji , Kanzawa Fumihiko , Inomata Motoko , Hoshi Akio
Published: February 2007

NK activity of human peripheral blood lymphocytes (PBL) for cells of the human myeloid line K562, and antibody‐dependent cellular cytotoxicity (ADCC) of PBL for cells of human lung adenocarcinoma line PC‐9 were determined by the human tumor clonogenic assay (HTCA). Incubation of K562 cells or anti‐PC‐9 serum treated PC‐9 cells with PBL before plating inhibited the formation of colonies of these tumor cells. The percent inhibition of tumor cell colony formation was dependent on the effector/target ratio, the incubation time before plating and, in the case of PC‐9 cells, on the dilution of anti‐PC‐9 serum. PBL activated with human T‐cell growth factor (TCGF), lympho‐kine‐activated killer (LAK) cells, significantly augmented the inhibition of colony formation of K562 cells, compared to the control lymphocytes. The increase in colony inhibition was dependent on the concentration of TCGF and the time of incubation of PBL with TCGF. The HTCA determining the colony inhibition of K562 cells incubated with LAK or PBL correlated with the 51Cr‐release assay (p < 0.001). The HTCA determining the colony inhibition of anti‐PC‐9 serum‐treated PC‐9 cells incubated with PBL also correlated with the 51Cr‐release assay (p < 0.001). We found that the NK activity and ADCC of lymphocytes on K562 and PC9 tumor lines could be detected with HTCA.
Distribution of laminin and collagen type‐IV in benign and malignant lesions of melanocytic origin
Authors: Natali P. G. , Nicotra M. R. , Bellocci M. , Cavaliere R. , Bigotti A.
Published: February 2007

Using antisera for two specific basement membrane (b.m.) antigens such as laminin and collagen type‐IV, together with efectron microscopy, we have shown that fully antigenic b.m. and morphologically typical basal lamina (b.l.) are associated with normal and transformed cells of the melanocyte lineage in different ways. Thus, while b.m. and b.l. surround individual choroidal melanocytes, intradermal nevic cells and cells of blue nevi are not detectable at the periphery of resting and proliferating epidermal melanocytes. They have a low degree of expression with a heterogeneous pattern of distribution in primary and meta‐static melanoma. This heterogeneity is present within single metastases and among autologous metastases. These findings indicate that the presence of b.m. can be an additional marker for cells of the melanocyte lineage and should be considered when applying ser‐ological means for the detection and control of neoplasms of melanocytic origin.
Inhibition of growth of certain human tumour cell lines by a factor derived from human fibroblast‐like cell lines. I. Demonstration by mixed culture and by use of cell washings
Authors: Wu Ke‐Fu , Pope John H. , Ellem Kay A. O.
Published: February 2007

The growth of 7/9 Burkitt's lymphoma (BL) cell lines and 5/6 human melanoma cell lines was inhibited by certain human fibroblast‐like cell lines, especially human myofibroblast cell lines. In contrast to BL lines, 3/5 lymphoblastoid cell lines were less susceptible and 3/4 human leukaemic lines were resistant. The effect was time‐ and concentration‐dependent. Co‐cultivation led to inhibition of tumour‐cell growth within 3 days, but after removal from the fibroblasts, washing, and resuspending in fresh medium some tumour cells recovered and growth improved. The inhibitory activity was also demonstrated with washings of fibroblast‐like cell lines, and was sensitive to pronase. The biological significance of this factor remains to be defined.
Sequential degradation of heparan sulfate in the subendothelial extracellular matrix by highly metastatic lymphoma cells
Authors: Bar‐Ner M. , Kramer M. D. , Schirrmacher V. , Ishai‐Michaeli R. , Fuks Z. , Vlodavsky I.
Published: February 2007

A highly metastatic variant (ESb) of a methylchol‐anthrene‐induced T lymphoma elaborates a heparan sulfate (HS) degrading endoglycosidase (heparanase) to a much higher extent than its non‐metastatic parental subline (Eb). Whereas a serum‐free medium conditioned by either subline contained a trypsin‐like serine protease, heparanase activity was detected only in the ESb‐conditioned medium (CM). ESb CM was incubated with a naturally produced, sulfate‐labelled subendothelial extracellular matrix (ECM) or with a soluble, high‐MW labelled proteoglycan first released from the ECM by incubation with Eb CM or with the partially purified ESb protease. Sulfate labelled degradation products were analyzed by gel filtration on Sephrose 6B. The optimal pH for degradation of ECM‐bound HS was 6.2 as compared to pH 5.2 for degradation of the soluble proteoglycan. Heparanase‐me‐diated degradation of both ECM‐bound and soluble HS was inhibited by heparin. Addition of either tryp‐sin, plasmin or to a lower extent, the purified ESb protease, stimulated between 5‐ and 20‐fold the ESb CM‐mediated degradation of ECM‐bound HS but had no effect on heparanase‐mediated degradation of the soluble proteoglycan. This stimulation was inhibited in the presence of heparin or protease inhibitors. These results indicate that both a protease and heparanase are involved in the ESb‐mediated degradation of ECM‐bound HS and that one enzyme produces a more accessible substrate for the next enzyme. This sequential cleavage is characteristic of degradation of a multimolecular structure such as the subendothelial ECM and hence cannot be detected in studies with its isolated constituents.
Muscarinic cholinergic receptors in pancreatic acinar carcinoma of rat
Authors: Taton Gérard , Delhaye Myriam , Swillens StéPhane , Morisset Jean , Larose Louise , Longnecker Daniel S. , Poirier Guy G.
Published: February 2007

The active enantiomer of tritiated quinuclidinyl benzilate (3H(‐)QNB) was used as a ligand to evaluate the muscarinic receptors. The 3H(‐)QNB binding characteristics of muscarinic cholinergic receptors obtained from normal and neoplastic tissues were studied to determine changes in receptor properties during neoplastic transformation. Saturable and stereospecific binding sites for 3H(‐)QNB are present in homogenates of rat pancreatic adenocarcinoma. The proportions of high‐ and low‐affinity agonist binding sites are similar for neoplastic and normal tissues. The density of muscarinic receptors is higher in neoplastic (200 femtomoles/mg protein) than in normal pancreatic homogenates (80 femtomoles/mg protein). The muscarinic binding sites of the neoplastic and fetal pancreas show similar KD values which are higher than those observed for normal pancreas.
Transforming potential of a retrovirus isolated from lung carcinoma of sheep
Authors: Safran N. , Zimber A. , Irving S. G. , Perk K.
Published: February 2007

A retrovirus isolated from experimentally induced sheep lung carcinoma (SPCTV) was propagated in chronically infected Himalayan tahr ovarian cells and in normal sheep lung cells. Follow‐up of infection of the cells with SPCTV showed the appearance of syncytium, plaque formation, partial recovery and the establishment of a chronic infection. Virus‐associated reverse transcriptase activity in the medium fluctuated but remained at a constantly high level at the stage of chronic infection. Stages of type‐C virus morphogenesis were demonstrated by electron microscopy. The viral genome was detected in both the nucleus and cytoplasm by in situ hybridization. Chronically infected cells formed colonies when plated in soft agar. Following subcutaneous inoculation of chronically infected cells (of fibroblast origin) into nude mice, lymphoid tumors developed at the site of inoculation and in vital organs.
Spontaneous and induced primary oncogenesis in natural killer (nk)‐cell‐deficient beige mutant mice
Authors: Haliotis Tina , Ball Judith K. , Dexter David , Roder John C.
Published: February 2007

Spontaneous tumor development and primary oncogenesis were compared in a large number of NK

Abbreviations: NK, Natural killer; s.c, subcutaneously; i.u., infectious units; d.f., degree(s) of freedom; DMBA‐LV, dime‐thylbenzanthracene‐induced murine leukemia virus; PVM, pneumonia virus of mice; Reo‐3, Reo virus 3; MHV, murine hepatitis virus; LCM, lymphocytic choriomeningitis virus; MEV, murine encephalomyelitis virus; MVM, minute virus of mice; BP, benzo[alpha]pyrene.

‐deficient, homozygous C57B1/6‐bg/bg mice and their NK normal, heterozygous + /bg littermate controls. In a group of 167 retired breeders followed for spontaneous tumors, the probability of survival for mice eventually dying with a tumor was greater for the NK‐competent, +/bg than for the NK‐deficient, homozygous C57BL/6‐bg/bg mice (p = 0.0019), although the higher overall incidence of tumors in the bg/bg group (48%) was not significantly different from that in the +/bg group (37%). In the bg/bg group the incidence of tumor death appeared to increase relatively sharply in the 25‐ to 29‐month age bracket compared to the fairly regular increase in incidence observed in the +/bg group. All the spontaneous tumors except 2 (discovered in +/bg mice) were classified histologically as widely disseminated malignant lymphomas. The other two were one squamous‐cell carcinoma and one sarcoma. A total of 73 bg/bg mice injected s.c. with benzo[alpha]pyrene (BP) had a higher overall incidence of tumors (81%) (rhabdomyosarcomas) than 138 +/bg mice (64%) and in the largest group (0.3 mg, n = 85) the bg/bg group developed tumors, at a higher incidence (p = 0.01) and with a shorter latency (p = 0.025) than the +/bg group. On the other hand, mice injected with dimethylbenzanthracene or given 4 weekly doses of 160 rads of gamma irradiation showed no difference in overall tumor incidence. In addition, mice injected with various doses of DMBA‐induced murine leukemia virus (DMBA‐LV) also showed no significant difference in tumor incidence. Others have reported that some of these treatments (DMBA, split‐dose irradiation) cause profound NK suppression, thereby reducing NK differences between the two groups of mice. These results suggest that a partial NK impairment in beige mutant mice early in life may lead to significantly greater rates of death with spontaneous malignant tumors late in life. Some primary oncogenesis treatments (BP) but not others (DMBA, split‐dose irradiation, leukemia viruses) cause tumors with a greater incidence and shorter latency in beige mice. The results suggest, but do not prove, that NK cells play a role in surveillance against spontaneously arising, and possibly some types of carcinogen‐induced, tumors.
The effect of murine interferon‐alpha/beta on an established rauscher murine leukemia virus‐induced erythroleukemia in balb/c mice
Authors: Herman R. A. C. P. , Trapman J.
Published: February 2007

Rauscher murine leukemia virus (R‐MuLV) induces a rapidly developing erythroleukemia in BALB/c mice. Previously, we have shown that mouse interferon‐α/β (Mu IFN‐α/β) applied shortly after virus inoculation efficiently inhibits the leukemic process (Hekman et al., 1981). Here we describe the effect of Mu IFN‐α/β on an established leukemia. Varying doses of Mu IFN‐α/β were injected over 3 days, starting 8 to 12 days after virus inoculation. The effect of Mu IFN‐α/β on the leukemic process was monitored by measuring the spleen weight, reverse transcriptase activity in the serum and, in selected experiments, by microscopic examination of sections of the spleen using standard histological and immunological staining techniques. Depending on the spleen weight at the start of its application (maximal about 450 mg), Mu IFN‐α/β caused a dramatic reduction in the number of virus‐infected erythroleukemic cells in the spleen. Also, R‐MuLV disappeared from the serum within 3 days. If Mu IFN‐α/β was injected into R‐MuLV‐infected mice with an already 10‐fold enlarged spleen, it could only stop further development of leukemia. Results obtained with crude Mu IFN‐α/β preparations were confirmed with absolutely pure Mu IFN‐β.
Hsv‐ and chemical carcinogen‐induced amplification of sv40 dna sequences in transformed cells is cell‐line‐dependent
Authors: Matz Bertfried , Schlehofer Jörg R. , Zur Hausen Harald , Huber Bernd , Fanning Ellen
Published: February 2007

Eleven simian virus 40‐transformed cell lines from 5 different species were tested for their ability to amplify integrated simian virus 40 DNA upon infection with herpes simplex virus type I or treatment with various chemical carcinogens. Four cell lines were positive only for virus‐induced gene amplification and two lines were positive for both carcinogen‐ and virus‐induced gene amplification. Individual cell lines were assayed for (1) the presence of an intact SV40 origin of replication, (2) the expression of a functional SV40 T‐antigen, and (3) permissivity to herpes simplex virus replication. These parameters were found to be positive in all 6 amplification‐competent cell lines. The ability of herpes simplex virus to amplify SV40 DNA sequences in transformed cells is greater than that of chemical carcinogens and can be suppressed by specific inhibitors of the herpes virus‐encoded DNA polymerase.
Tumor‐associated desmoplasia in the rabbit mesentery characterized by morphological, biochemical and cytophotometric methods
Authors: Haemmerli G. , Müller‐Glauser W. , Bruckner P. , Hauser‐Urfer I. , Sträuli P.
Published: February 2007

Intraperitoneal implantation of V2 carcinoma cells in the rabbit leads to invasion of the mesentery and to structural tissue alterations which are concomitantly of a destructive and a desmoplastic type. In this report, we describe the desmoplastic changes which are characterized by the increased formation of collagen and of proteoglycans resulting in an increased thickness of the membrane. Biochemical data indicate that the total amount of collagen increases with time after implantation, whereas the relative amount per unit of dry weight, as well as the contributions of type I (15–25%) and type III (6–8%), stay within the same range. The increased synthesis of extracellular matrix is accompanied by a change in the appearance of the fibroblasts which now show the morphologic features of synthesizing cells. Also, an appreciable number have entered the S‐phase. We propose that the desmoplastic changes are tumor‐associated, since implantation of epithelial cells from normal rabbit liver does not result in similar alteration. Our findings are discussed in view of the role played by tumor and/or host cells in the increased production of extracellular matrix, of possible factor(s) elaborated by the tumor cells, and of the general significance of desmoplastia for tumor spread.
Monoclonal antibody production to a B16 melanoma associated antigen
Authors: Tomita Yasushi , Montague Paul M. , Hearing Vincent J.
Published: February 2007

Eight different rat hybridoma cell lines have been produced which secrete MAbs

Abbreviations: BSA, bovine serum albumin; ELISA, enzymelinked immunosorbent assay; MSA, murine serum albumin; PBS, phosphate‐buffered saline; MAb, monoclonal antibody; s.c., subcutaneous.

identifying the melanoma‐specific B700 antigen. This antigen has been previously shown to have significant sequence homology to the mammalian serum albumins, but antibodies specifically raised against B700 cross‐react only with murine albumin. Each of these MAbs has been shown by Western immunoblotting to recognize an independent epitope on the B700 protein; it is concluded not only that the B700 protein is a highly immunogenic antigen, but also that the sequence homologies between the B700 antigen and murine albumin must occur extensively throughout the molecule.
Effects of cortisone with and without heparin on angiogenesis induced by prostaglandin e1 and by s180 cells, and on growth of murine transplantable tumours
Authors: Ziche Marina , Ruggiero Marco , Pasquali Franca , Chiarugi Vincenzo P.
Published: February 2007

Cortisone acetate, locally applied in sustained‐release pellets, is effective in inhibiting angiogenesis induced by prostaglandin EI in the rabbit cornea. The inhibitory effect of cortisone is not increased by addition of heparin. Similar results were obtained with angiogenesis induced by S180 cells. The effects of cortisone with and without heparin were also studied on 5 transplantable murine tumours: 3 variants of B16 melanoma, Lewis lung carcinoma and fibrosarcoma M4. The effect of cortisone in slowing the growth rate of tumours was modestly potentiated by heparin, but no regression of the tumour mass occurred.
Selective cytotoxicity of murine monoclonal antibody LAM2 against human small‐cell carcinoma in the presence of human complement: Possible use for in vitro elimination of tumor cells from bone marrow
Authors: Stahel Rolf A. , Mabry Mack , Sabbath Kert , Speak Jonathan A. , Bernal Samuel D.
Published: February 2007

LAM2 is a murine IgM monoclonal antibody (MAb) which binds strongly to the cell membrane of human lung small‐cell carcinoma (SCC) and squamous‐cell carcinoma but not to normal bone‐marrow cells. The cytotoxicity of this antibody in the presence of human complement was investigated in vitro by chromium release and clonogenic assays. The optimal treatment conditions included incubation with antibody for 30 min at 37°C followed by 3 additions of human complement 30 min apart. Cell lysis ranged from 94 to 98% in 4 SCC cell lines at antibody dilutions of 1: 100: a lower level of lysis (60%) occurred in a lung squamous cell carcinoma cell line. The cytotoxic effect was strictly complement‐dependent. No cytotoxic effect was seen with other human cell lines including lung adenocarcinoma, lung large‐cell carcinoma, myeloid leukemia, and lymphoblastic leukemia. No lysis was seen with nucleated marrow cells from healthy volunteers. Normal marrow cells in excess did not inhibit SCC cell lysis. Incubation with antibody and complement resulted in a 100‐fold reduction of colony formation of SCC cells, but did not reduce the number of colonies of marrow‐cell precursors, including CFU‐GEMM, BFU‐E, and CFU‐C. The selective cytotoxicity of LAM2 antibody to SCC, but not to normal bonemarrow cells, suggests that this antibody may be useful for in vitro elimination of SCC cells from the bone marrow.
Oesophageal cancer studies in the caspian littoral of Iran: Some residual results, including opium use as a risk factor
Authors: Ghadirian P. , Stein G. F. , Gorodetzky C. , Roberfroid M. B. , Mahon G. A. T. , Bartsch H. , Day N. E.
Published: February 2007

A study was conducted in northern Iran in areas of very high, high and moderately low incidence of oesophageal cancer. Morphine metabolites in urine as an indicator of opium use, and a variety of nutritional and biochemical measures, including salivary antipyrine half‐life, were determined in households with a case of oesophageal cancer and in control households. Results on 1,590 individuals showed that the prevalence of appreciable levels (≤ 1μ/ml) of urinary morphine metabolites was much higher in areas of high and very high incidence of oesophageal cancer than in low‐incidence areas, particularly for those under age 50 years in both sexes, where a 6‐fold difference was seen. Members of households with a case of oesophageal cancer had a higher prevalence of positive urinary morphine metabolite findings than members of control households from the same village. Salivary antipyrine half‐life (AP‐T 1/2), as a measure of hepatic drug metabolizing capacity, was estimated in 120 subjects. No relationship was seen with use of opium, region of residence, case‐control status of household, age or sex. Experimental studies have meanwhile identified mutagenic substances with carcinogenic potential in opium pipe scrapings, used widely in the study region, opium pyrolysates and morphine pyrolysates. Our data give additional support to the hypothesis that opium use, in the form of its pyrolysates, is one of the factors involved in the aetiology of oesophageal cancer in the region.
Use of novel chemical supplements in the establishment of three human malignant lymphoma cell lines (NU‐DHL‐1, NUDUL‐1, and NU‐AMB‐1) with chromosome 14 translocations
Authors: Epstein Alan L. , Variakojis Daina , Berger Carol , Hecht Barbara K.
Published: February 2007

Three new cell lines have been established from patients with malignant lymphoma utilizing a human diploid feeder layer, pooled human serum, and the chemical supplements L‐cysteine, iron‐saturated transferrin, and bathocuproine disulfonate, a copper chelator. After a short period of growth, the 3 cell lines were successfully weaned from the feeder layers but continued to require human serum and the chemical supplements for up to 9 months of culture. The cell lines are currently grown in RPMI–1640 medium and fetal calf serum without further supplementation. The NU‐DHL‐I cell line was established from the involved lymph node of a 73‐year‐old White male with diffuse large‐cell lymphoma. The cell line expresses cytoplasmic IgM/lambda heavy and light chains, is Epstein‐Barr virus (EBV)‐negative, and is positive for several B‐cell markers, indicating that it is derived from a mature‐B‐cell neoplasm. The NU‐DUL‐1 cell line was established from the cerebrospinal fluid of a 42‐year old White male with undifferentiated lymphoma, non‐Burkitt's type, who initially presented with a mediastinal mass and had subsequent involvement of the central nervous system. The cell line is EBV‐negative, but surprisingly it is positive for early B‐cell markers. The NU‐AmB‐1 cell line was established from the abdominal mass of a 12‐year‐old Hispanic male with undifferentiated lymphoma, Burkitt's type. The cell line is EBV‐positive and expresses early B‐cell markers. All 3 cell lines are aneuploid or pseudodiploid and contain chromosome 14q+ abnormalities including a newly described complex translocation t(?; 1; 8; 14) in the NU‐AmB‐1 cell line. The establishment of these cell lines was made possible by refinements in the cell culture of the human malignant lymphoma. The availability of well‐characterized lymphoma cell lines with specific chromosomal translocations will aid molecular and cellular studies designed to identify the biological significance of genomic rearrangements.
Gangliosides in human chronic lymphocytic leukaemia
Authors: Anh‐Tuan Nguyen , Pick Judit , Vajda János , Harsányi Veronika , Hollán Susan R.
Published: February 2007

Gangliosides were analysed by elaborated overpressured thin‐layer chromatography (OPTLC) in 6 cell samples from 5 patients with chronic lymphocytic leukaemia (CLL) of B‐cell origin, and 4 individual lymphocyte preparations from normal blood donors. The principal difference in the ganglioside profile between these two counterparts appears to be the presence of GD3 and the predominance of the less polar compounds (GM3, GM2, GM1) in CLL cells. Qualitatively, GD3, accounted for about 5. 5% of the total CLL gangliosides, whereas it was not detectable in normal lymphocytes. Quantitatively, GM3 constituted more than 81% of the total CLL gangliosides, a proportion more than twice as high as that found in normal lymphocytes. Three other minor gangliosides were isolated from CLL cells; these were shown to be GM2 (trace amounts), GM1 (7. 7%), and GD1 (4%). The expression of individual gangliosides varied greatly among the various CLL samples obtained from different patients, and even from the same patient, if examined at different times. No gangliosides were found in the supernatant collected from CLL cells subjected to a temperature shift (0°C to 37°C) or in the cell‐free medium harvested from the CLL cells kept in overnight culture.
N‐ras gene activation in the rd human rhabdomyosarcoma cell line
Authors: Chardin Pierre , Yeramian Patrick , Madaule Pascal , Tavitian Armand
Published: February 2007

A transforming N‐ras allele was molecularly cloned from the RD human rhabdomyosarcoma cell line, and the nature of its activation studied. Construction of chimeric recombinants between the RD‐transforming allele and a normal human allele enabled us to localize the alteration responsible for the activation to the second exon of the N‐ras gene. The nucleotide sequence of this exon, when compared to that of the normal allele, revealed a single difference at the 61st amino acid position of the encoded protein; the CAA codon for glutamine in the normal allele was mutated to a CAT codon for histidine in the RD‐transforming allele. This result is the first description of a histidine replacing glutamine in the 61st position and provides further evidence that the 61st amino acid is one of the preferential sites for N‐ras activation.
Cell DNA content — Correlation with clonogenicity in the human tumour cloning system (HTCS)
Authors: Verheuen R. H. M. , Feitz W. F. J. , Beck J. L. M. , Debruyne F. M. J. , Vooys G. P. , Kenemans P. , Herman C. J.
Published: February 2007

Thirty‐six ovarian and renal‐cell tumours were analyzed by flow cytometry (FCM) for DNA content and in parallel were assayed for colony formation in a human tumour cloning system (HTCS). While 15/19 (79%) tumours with an abnormal (aneuploid) DNA stemline formed colonies in the HTCS, only 2/17 (12%) of diploid tumours formed colonies. All samples contained tumour cells as assessed by routine cytological examination. The capacity to form colonies in the HTCS was not correlated in these tumours with grade or stage of disease or tumour type. The level of aneuploidy expressed as the FCM DNA index did not correlate with the cloning efficiency in HTCS. These findings suggest that tumour growth in the HTCS reflects a biologically important potential, related in at least some turnours to an abnormal DNA stemline.
Phenotypic interconversion of B16 melanoma clonal cell populations: Relationship between metastasis and tumor growth rate
Authors: Stackpole Christopher W. , Fornabaio Donna M. , Alterman Andrea L.
Published: February 2007

Three distinct dissemination‐related phenotypes have been recognized in clones of the mouse B 16 melanoma based on in vivo behavior: metastatic (spontaneously disseminating to the lungs from solid tumors), colonizing (capable of forming tumor colonies in the lungs following intravenous injection), and null (tumorigenic but non‐metastatic and non‐colonizing). From a progenitor null clone, G3, subclones that became phenotypically diversified in vitro (metastatic G3.5 and null G3.15) and in vivo (metastatic G3.12 and colonizing G3.26) were derived. During long‐term culturing, G3 cells became metastatic and then lost that activity, G3.5 and G3.12 cells gradually lost metastatic activity, and G3.26 cells became slightly metastatic and non‐colonizing. Subclone G3. I5 became highly metastatic after a single subcutaneous (s. c.) tumor passage. In aged mice, and in young mice injected with incompletely‐tumorigenic cell doses, G3 and G3.26 s. c. tumors were metastatic, but cells cultured from those tumors or metastases were non‐metastatic when tested in young mice at standard highly‐tumorigenic cell doses. The behavior of G3.5 and G3.12 tumors was not altered in aged mice or when tumors were initiated with small cell inocula. Analysis of growth characteristics associated with these phenotypic interconversions indicated that lung‐colonizing potential was directly related to the ability of the cells to grow as multicell colonies in 0.3% agar, and that metastatic activity was expressed by tumors that grew at moderate rates. In young mice receiving standard cell doses, G3.5 and G3.12 tumors inherently grew at that rate, whereas G3 and G3.26 tumors grew more rapidly and G3.15 tumors grew more slowly. Regardless of inherent phenotype, all clones were capable of expressing metastatic activity, at least transiently, as tumor growth was altered to moderate rates. Expression of metastatic behavior might, therefore, be regulated to some extent by tumor growth characteristics.
Clonal analysis of cytotoxic T‐lymphocyte response to autologous human metastatic melanoma
Authors: Anichini Andrea , Fossati Giuseppe , Parmiani Giorgio
Published: February 2007

Peripheral blood lymphocytes (PBL) from a melanoma (Me) patient, previously shown to be unable to react against the autologous tumor (Me 28) after mixed lymphocyte‐tumor culture (MLTC), were cultured in vitro with the autologous tumor in MLTC and/or with IL‐2‐containing supernatants. T‐cell clones were then obtained by limiting dilution and by micromanipulation. Eleven clones, selected for autologous tumor (Auto‐Tu) cytotoxicity, were tested for specificity on a panel of 17 cell cultures of normal and neoplastic origin, revealing a complex spectrum of lytic activities. Three groups of clones could be identified depending on the patterns of cytotoxicity. One clone (B11.12) lysed Me28 and expressed a borderline reactivity against one allogeneic Me. A second group of clones (A4, A4.18, H10, E12, C9) lysed the Auto‐Tu and allogeneic Me. The last group of clones (A4.2, A4.3, A4.4, A7, B7) expressed a broader pattern of reactivity with significant cytotoxicity against targets of different histologic origin. Furthermore, the second and third groups of clones lysed K562 while B11.12 did not. The Auto‐Tu‐restricted reactivity of clone B11.12, confirmed by a further test on 13 allogeneic Me and on autologous IL‐2 cultured lymphocytes, suggests the recognition of antigenic structures preferentially expressed on Me28. Blocking studies, performed with monoclonal antibodies (MAb), revealed that an anti‐HLA class 1 MAb (w6/32), but not two anti‐DR MAbs (L243, D1.12), could inhibit the cytotoxic activity of clones B11.12 on Me28. A significant blocking effect by w6/32 on Me28 was achieved also with clones A4.4 and H10 but not with clones A4.2, A4.3 and A7. The phenotype of all clones was T3+, T4−, T8+, HNK‐1−, suggesting that the anti‐tumor effectors were of the T‐cell lineage. Taken together, these data indicate that it is possible to isolate anti‐tumor CTL‐clones after MLTC from a PBL population of a metastatic melanoma patient. Furthermore, we present evidence suggesting a role of class‐1 antigens in the interaction of some cloned effectors with the autologous tumor target.
Familial and environmental interactions in bladder cancer risk
Authors: Kantor Arlene F. , Hartge Patricia , Hoover Robert N. , Fraumeni Joseph F.
Published: February 2007

In a population‐based study of 2,982 bladder cancer patients and 5,782 controls in 10 geographic areas of the United States which was designed to assess the role of environmental risk factors, information was also obtained on the history of urinary tract cancer in first‐degree relatives. A family history of urinary tract cancer significantly elevated the risk of bladder cancer [relative risk (RR) = 1.45], with higher risks observed among patients under age 45. The risks of bladder cancer associated with positive family history were generally higher among persons with suspected environmental exposures, particularly heavy cigarette smoking (RR = 10.7 among those who smoked 3 or more packs per day). Further studies of bladder cancer should incorporate biochemical and genetic probes to assess mechanisms of familial susceptibility and interactions with environmental factors.
Combined recombinant human interferon alpha2 and cytotoxic agents studied in a clonogenic assay
Authors: Welander Charles E. , Morgan Timothy M. , Homesley Howard D. , Trotta Paul P. , Spiegel Robert J.
Published: February 2007

A human tumor clonogenic assay has been used to test the antiproliferative effect of recombinant human leukocyte interferon alpha2 alone and in combination with each of 8 cytotoxic agents. Cell lines derived from 6 human tumors and primary tumor cells from 13 patients have been used in these clonogenic assay studies. Results show that interferon as a single agent causes insignificant reduction in tumor cell colony survival if the short‐term 1‐hr cell exposure method is used; only high concentrations of interferon used in continuous cell exposure in the clonogenic assay can demonstrate a reduction in colony survival to below 50% of control values. Combinations of interferon with either doxorubicin or cisplatin frequently show additive and occasionally synergistic antiproliferative effects on tumor cell colony formation. Variations in drug concentrations and sequencing of drugs have been tested, showing that optimal antiproliferative effects of combined interferon and doxorubicin are realized when maximal concentrations of interferon and prolonged cell exposure time of both interferon and doxorubicin are employed. Combinations of interferon and doxorubicin tested in the clonogenic assay demonstrate cytotoxicity superior to that of either agent tested alone.
Oncogene expression in human tumors
Authors: Tatosyan A. G. , Galetzki S. A. , Kisseljova N. P. , Asanova A. A. , Zborovskaya I. B. , Spitkovsky D. D. , Revasova E. S. , Martin P. , Kisseljov F. L.
Published: February 2007

The expression of 9 oncogenes in primary tumors and in human tumors passaged in nude mice was tested: a total of 28 tumor types was analyzed. Oncogenes src, fps, and mos were not expressed in any of the tumors tested but oncogene myc was transcribed in most of the tumors and myc was overexpressed in 3 tumors passaged in nude mice (Ewing sarcoma, large intestine carcinoma and kidney carcinoma) and in primary fibrous histiocytoma. Enhanced transcription of ras and fos genes was observed nonspecifically in different tumors. Oncogene sis was activated specifically in metastases of different tumors in lymph nodes.
Decreased calcium dependence of lymphoblastoid cell lines compared with burkitt lymphoma cell lines
Authors: Parsons P. G. , Moss D. J. , Morris C. , Musk P. , Maynard K. , Partridge R.
Published: February 2007

The extracellular calcium level required for proliferation was compared in B lymphoid cell lines from various sources by determining the calcium concentration at which long‐term proliferation was inhibited by 50% (CaPD50). Fourteen Burkitt lymphoma (BL) lines had a mean CaPD50 of 44 ± 28 μM whereas 45 lymphoblastoid cell lines (LCLs) obtained by in vitro transformation of B lymphocytes with Epstein‐Barr virus (EBV) had a mean CaPD50 of 3.6 ± 1.8 μM. This difference applied also to autologous BL lines and LCLs established from the same patient. The decreased calcium requirement of virally‐transformed compared with tumour‐derived cell lines therefore appears to be a universal phenomenon in mamalian cells. Within the BL group, no correlation was found between the calcium requirement for proliferation and presence or absence of the EBV genome. Arrest of BL lines and LCLs occurred in the G1 phase of the cell cycle and was readily reversed by additional of calcium to the medium. One anomalous LCL was found which showed a high CaPD50 (43 ± 6 μM) and accumulated in both G1 and G2. These results, in combination with a previous study of EBV transformation in vitro, indicate that the calcium dependence of B lymphocytes generally decreases in the following order: normal cells > BL cells = early stage transformation > LCL. The 2 transformed phenotypes thus distinguished in human lymphoid cells may offer unique opportunities for defining the status and expression of EBV in vitro and in vivo.
Mononuclear cells infiltrating human mammary carcinomas: Immunohistochemical analysis with monoclonal antibodies
Authors: Hurlimann Jean , Saraga Puica
Published: February 2007

Breast carcinomas were examined by the immunoperoxidase technique using antisera specific for lymphocyte subsets, monocytes, NK cells and major histocompatibility antigens (HLA‐A,‐B,‐C; la‐like). Sixty‐four per cent of the patients had a moderate or strong mononuclear cell infiltration, 77% of the patients without mononuclear cell infiltration had receptors for estrogens as compared to 51% of the patients with infiltration. The majority of the infiltrating mononuclear cells were T cells; generally the OKT8, cells were predominant. The Leu 3A/OKT8 cell ratio was not related to histological type, tumor size, age of the patient or presence of metastases. Some of the T cells had the la antigen and were thus probably activated. The B cells were either absent or less numerous than the T cells. There was no relation between their distribution and the various parameters studied. A few monocytes were heterogeneous according to their markers (OKM I and acid phosphatase). In 6 cases only there was a strong infiltration of mononuclear cells positive for acid phosphatase. The number of the natural killer cells was also low. Only a few mononuclear infiltrating cells had receptors for transferrin. There was a positive correlation between the inflammatory infiltration and the presence of HLA class‐1 antigens on tumor cells. Some of the antisera specific for lymphocyte subsets also stained the breast carcinoma cells. The great variations in the subsets of mononuclear cells in breast carcinomas may correspond to various systems of defense against neoplasm.
Monoclonal‐antibody‐defined human lung tumour cell‐surface antigens
Authors: Reeve Julie G. , Wulfrank Denis A. , Stewart Jonathan , Twentyman Peter R. , Baillie‐Johnson Hugo , Bleehen Norman M.
Published: February 2007

A panel of 3 monoclonal antibodies (MAbs) directed against human lung tumour cell‐surface antigens has been produced following immunizations with the established small‐cell lung cancer (SCLC) cell line, NCl‐H69, and with another SCLC cell line, COR‐L32, recently derived from clinical material. One MAb, B10/12, reacted strongly with SCLC, immunoprecipitated a protein having an MW of 100kd and failed to react significantly with non‐small‐cell lung cancer (NSCLC) in radioimmunoassay and in an immunohistochemical assay. MAbs E10/5 and 2G3 reacted extensively with SCLC but also showed significant reactivity with NSCLC. MAb E10/5 immunoprecipitated a protein with an MW of 80kd but no appreciable protein was specifically precipitated by MAb 2G3. Unlike MAb 2G3, both MAbs B10/12 and E10/5 reacted strongly with selected neuroblastomas whereas only MAbs 2G3 and E10/5 reacted significantly with melanoma. All 3 MAbs reacted with breast carcinomas. Other non‐pulmonary tumours thus far examined failed to react with the MAbs in radioimmunoassay or immunohistochemical assay. Immunocytochemistry and the use of viable cells in radioimmunoassay confirmed that the antigenic determinants recognized by these MAbs were surface located.
In vitro cisplatin effects on phagocyte functions
Authors: Fumarulo Ruggiero , Riccardi Silvana , Pantaleo Romeo , Coluccia Mauro , Giordano Domenico , Aresta Michele
Published: February 2007

The effects of cis‐diamminedichloroplatinum (11) (cisplatin), a drug used in cancer chemotherapy, on the oxidative metabolism, endocytosis, chemotaxis and exocytosis of guinea‐pig polymorphonuclear leukocytes were studied. All these functions were negatively influenced, but the same effect (50% inhibition) was observed at different drug concentrations (3 × 10−5 M for chemotaxis, 10−4 M for O2 consumption by FMLP and β‐glucuronidase release, 10−3 M for O2 consumption by PMA and for zymosan engulfment). The effects of the drug can be explained by its ability to bind to membrane proteins, essentially to ‐SH groups.