bluuebunny/crossref_metadata_2025 huggingface dataset:
Combination tumor‐immunotherapy with recombinant tumor necrosis factor and recombinant interleukin 2 in mice
Authors: Nishimura Takashi , Ohta So , Sato Naoko , Togashi Yuji , Goto Makiko , Hashimoto Yoshiyuki
Published: February 2007

Recombinant human tumor necrosis factor (r‐TNF) inhibits growth of various mouse tumor cell lines both in vitro and in vivo. Treatment of established tumor nodules with intratumoral (i.t.) injection of r‐TNF caused hemorrhagic necrosis of tumor and temporary disappearance of tumor mass. However, a small number of tumor cells remained and later formed fresh nodules. In striking contrast, combination therapy with r‐TNF and recombinant human interleukin‐2 (r‐IL‐2) resulted in a marked inhibition of regrowth of tumor cells. More than 60% of MBL‐2‐bearing mice were completely cured of tumor by treatment with r‐TNF and r‐IL‐2. Cured mice could also reject rechallenged MBL‐2 lymphoma cells, indicating the generation of anti‐tumor effector cells in vivo. However, lymphocytes obtained from mice cured of MBL‐2 showed no significant in vitro cytotoxic activity against MBL‐2 lymphoma cells. In contrast, in vitro sensitization of spleen cells from cured mice with mitomycin‐C‐treated MBL‐2 lymphoma cells resulted in the generation of cytotoxic cells against MBL‐2 lymphoma cells. Moreover, spleen cells from mice cured of MBL‐2 by treatment with r‐TNF and r‐IL‐2 revealed a strong anti‐tumor activity upon in vivo neutralization tests. These results strongly suggest that tumor‐bearing mice can acquire systemic immunological memory after combination therapy with r‐TNF and r‐IL‐2.
Phenotype versus immunoglobulin and T‐cell receptor genotype of Hodgkin‐derived cell lines: Activation of immature lymphoid cells in Hodgkin's disease
Authors: Falk Martin H. , Tesch Hans , Stein Harald , Diehl Volker , Jones David B. , Fonatsch Christa , Bornkamm Georg W.
Published: February 2007

Three Hodgkin‐derived cell lines (L428, L540, and CO) were studied for rearrangements and expression of immunoglobulin and T‐cell receptor genes, and their genotype was compared to the phenotype. As far as the genotype is concerned, all 3 cell lines have characteristics of lymphoid cells; L428 of B, and L540 and CO of T‐cell origin. L428 cells have one Ig heavy chain allele rearranged to Cγ and transcribed into RNA, while the second is deleted. Furthermore, L428 cells show an unusual immunoglobulin kappa light chain gene rearrangement involving deletion of the kappa constant gene in one allele, while the remaining kappa and lambda loci are in germ‐line configuration. L540 and CO have, in contrast to L428 cells, the immunoglobulin genes in germline and T‐cell receptor genes rearranged. The T‐cell receptor β and γ genes are rearranged in both L540 and CO, whereas a rearrangement in the a locus was detected in L540 cells only. RNA of the size of functional β chain transcripts was found in CO cells and of the size of functional α chain transcripts in L540 cells. All 3 cell lines are classified as immature lymphoid cells with respect to the limited expression of B‐and T‐cell antigens, respectively, and to the incomplete expression of their antigen receptor. The immaturity of lymphoid differentiation contrasts with the expression of activation antigens, i.e. Ki‐1, Ki‐24, HLA‐DR, and IL‐2 receptor. The immaturity of the cells excludes the possibility that the cells were activated along the physiological pathway, i.e. by interaction of the cell with antigen. The results obtained on the cell lines are in accordance with in vivo studies and suggest that Hodgkin and Sternberg‐Reed cells are immature lymphoid cells which are activated by a still unknown mechanism.
Epidemiology of respiratory cancers related to nickel mining an refining in New Caledonia (1978–1984)
Authors: Goldberg M. , Goldberg P. , Leclerc A. , Chastang J. F. , Fuhrer R. , Brodeur J. M. , Segnan N. , Floch J. J. , Michel G.
Published: February 2007

The incidence rate of respiratory (lung and upper respiratory tract) cancer related to the nickel industry was sudied in the male population of New Caledonia over a 7‐year period (1978–1984). The findings show no excess incidence of lung or upper respiratory tract (larynx, pharynx, nasal cavities) cancer cases in the population of nickel workers compared with the rest of the male population in New Caledonia. This result was corroborated by a case‐control study which does not indicate any particular role of hazards specific to the nickel mining and refining industry. Our findings, which may be specific 1 or the type of ore processed in New Caledonia, must nevertheless be confirmed by extending the study to a total of 10 years (1978–1987). The incidence rate of male respiratory cancer in New Caledonia was also compared to that of cancers of this type reported in certain regions of industrialized countries (Australia, France, U.K., USA). Such findings are very similar to those observed in New Caledonia, thereby confirming a predominant role of tobacco and alcohol consumption in a territcry which, despite certain characteristics of a developing country, has life‐style habits similar to those of industrialized countries.
Specific uptake of alpha‐fetoprotein by malignant human lymphoid cells
Authors: Laboida Jorge , Naval Javier , Allouche Michèle , Calvo Miguel , Georgoulias Vassilis , Mishal Zohair , Urkl José
Published: February 2007

We have studied the ability of human B‐and T‐lymphoblastcid cell lines, as well as of peripheral lymphoid cells from leukemia patients, to take up alpha‐fetoprotein (AFP) and other serum proteins. Two technical approaches have been employed, both using fluorescent protein derivatives (FITC‐proteins): microscopic examination of labelled cells using epifluorescent illumination and quantitation of endocytosed proteins by fluorescence‐activated cell sorting (FACS). Compared to human resting T‐lymphocytes, all T‐and B‐cell lines tested exhibited positive staining for fluoresceinated AFP and transferrin (Tf) and a significant increase, up to 100‐fold, of the number of AFP and Tf molecules endocytosed per cell. Labeling was prevented or strongly diminished by a 100‐fold excess of unlabelled protein. Preliminary results with peripheral lymphoid cells harvested from leukemia patients showed the presence of AFP‐and Tf‐positive cells in the blood of all patients examined. Intensity of labelling was related to the type of leukemia cells and/or the degree of cell maturation. Host cell lines exhibited positive staining for alpha2‐macroglobulin (α2M) and also, to a lesser extent, for serum Vitamin D3 binding protein (DBP). In contrast, no labelling was observed with FITC‐serum albumin (FITC‐Alb) or FITC‐ovalbumin (FITC‐OVA). Comparative uptake of several FITC‐protcins by a single cell population revealed significant quantitative and qualitative differences.
Immunological quantitation of phospholipid/CA2+‐dependent protein kinase of human mammary carcinoma cells: Inverse relationship to estrogen receptors
Authors: Borner Christoph , Wyss Rudolf , Regazzi Romano , Eppenberger Urs , Fabbro Doriano
Published: February 2007

The amounts of phospholipid‐and Ca2+‐dependent protein kinase (PKC) of various human mammary tumor cells containing (ER+) or lacking (ER‐) estrogen receptors were estimated by quantitative immunoblotting. According to several criteria the polyclonal anti‐PKC antibody raised in rabbits against porcine brain PKC specifically recognizes an 80‐kDa polypeptide on immunoblots. This 80‐kDa PKC presumably represents the autophosphorylated form of the holoenzyme. Immunological quantitation of PKC revealed that the levels of immunodetectable PKC varied widely among the various human mammary carcinoma cell lines but closely matched the amounts determined by enzyme activity and phorbol ester binding in the respective cell line. The largest amounts of immunodetectable PKC were found in the ER‐human mammary tumor cells (0.5 to 1.5 μg PKC/mg of cytosolic protein). These data indicate that ER‐human mammary carcinoma cell lines express significantly higher levels of PKC than their estrogen‐receptor‐containing counterparts.
Deferent behaviour of normal and neoplastic cells cultured in vitro in the presence of catalase and superoxide dismutase
Authors: Liotti F. S. , Bodo M. , Menghini A. R. , Guerrieri P. , Mariucci G. , Bruschelli G.
Published: February 2007

Chicken embryo fibroblasts and hepatocytes were studied in the presence of catalase and superoxide dismutase in order to establish whether these enzymes had the capacity to favour cell multiplication as previously shown for in vitro tumour ascites cells (ATP C+). The results indicate that, unlike ATP C+ cells, both fibroblasts and hepatocytes are inhibited in their multiplication by superoxide dismutase. Similar effects are exerted on hepatocytes by catalase, whereas the multiplication of fibroblasts is favoured by high doses of this enzyme. Enzyme determinations revealed high levels of catalase and superoxide dismutase in hepatocytes, whereas both enzymes were poor in fibroblasts and ATP C+.
Lysis of tumor cells by the retargeting of murine cytolytic T lymphocytes with bispecific antibodies
Authors: Barr I. G. , Macdonald H. R. , Buchegger F. , Von Fliedner V.
Published: February 2007

The specificity and efficiency of tumor‐cell lysis by the retargeting of murine cytolytic lymphocytes (CTL) with bispecific antibodies was examined. Bispecific antibodies (also known as heteroaggregated or hybrid antibodies) were produced by the chemical coupling of monoclonal antibodies (MAbs) against H‐2 antigens and the murine T‐cell receptor (TCR). Murine tumor cell lines which expressed on their plasma membrane an antigen reactive with one component of the bispecific antibody were efficiently lysed in the presence of polyclonal murine CTL. CTL capable of lysis were generated by stimulating spleen or lymph‐node cells with ConA and IL‐2, while unstimulated cells or cells incubated only with IL‐2 showed no lysis of target cells with bispecific antibodies. Furthermore, the lysis of target cells by bispecific antibodies and CTL did not cause the lysis of bystander cells (cells not expressing an antigen recognized by the antibody which are mixed with the target cells). The efficient CTL‐mediated lysis observed with these antibodies makes this a promising approach for the immunotherapy of human cancer.
Interleukin‐2 activated human killer lymphocytes: Lack of involvement of interferon in the development of IL‐2‐activated killer lymphocytes
Authors: Damle Nitin K. , Doyle Laura V.
Published: February 2007

When cultured with native or recombinant interleukin‐2 (IL‐2) human small agranular lymphocytes acquire the ability to kill various tumor targets. The development of these IL‐2 activated killer (IAK) cells, also known as LAK, is observed in the absence of antigen or mitogen. Interferons are known to augment the lytic effects of natural killer cells and cytolytic T lymphocytes. Our study was undertaken to examine the effect of human α, β, and γ interferons on the induction and the effector phase of IAK function. When cultured with small lymphoid cells IFN alone did not induce anti‐tumor cytolytic activity in those lymphocytes. Despite their known anti‐proliferative effects, none of the 3 IFN species at any concentrations tested inhibited the development of IAK activity when present during the entire culture period. IFN neither inhibited nor augmented the development of IAK cells under suboptimal conditions. Furthermore, activation of IAK cells was not affected by the presence of neutralizing antibodies to either α or γ IFN. Post‐activation exposure of IAK cells to IFN also failed to either augment or inhibit their lytic activity. Thus, neither endogenously generated nor exogenously added IFN had any effect on the IAK system, in contrast to their augmenting effects on NK cells and CTL.
Lymphogenous and hematogenous metastasis of Lewis lung carcinoma in the mouse
Authors: Weiss L. , Ward P. M.
Published: February 2007

Quantitative and temporal comparisons were made between lymphogenous and hematogenous metastasis in the non‐immunogenic model system of mice bearing transplanted 3LL tumors in their hind feet. The experimental evidence indicates that cancer cells disseminate from clinically detectable primary cancers by non‐exclusive routes in the bloodstream and in the lymphatics. Following a delay of approximately 2 weeks after injection of 3LL cancer cells into the foot, local lymph‐node micrometastases occur, together with the first appearance of overt hematogenous metastases in the lungs. The anatomic extent of lymph‐node involvement, determined by bioassays of orthotopic grafts, of ipsilateral popliteal, inguinal and lumbar nodes, provides an accurate indicator of hematogenous metastasis, even though lymphogenous and hematogenous metastasis are operationally independent in this tumor/host system.
B‐cell lymphomas of high‐grade malignancy frequently lack HLA‐DR, ‐DP and ‐DQ antigens and associated invariant chain
Authors: Momburg Frank , Herrmann Barbara , Moldenhauer Gerhard , Möller Peter
Published: February 2007

The expression of HLA‐DR, HLA‐DP and HLA‐DQ antigens was studied in an unselected series of 66 B‐cell lymphomas by means of immunohistology using monoclonal antibodies against non‐polymorphic determinants in a sensitive immunoperoxidase technique. In addition, the expression of the MHC class‐II antigen‐associated invariant chain (li) was examined, The tumors were classified according to the Kiel classification, 30 being of high‐grade and 36 being of low‐grade malignrancy. Only 6 lymphomas of high‐grade and 16 lymphomas of low‐grade malignancy showed coordinate strong expression of all HLA class‐II antigens and invariant chain as observed in the normal peripheral B cell. Six further tumors of high‐grade and 8 tumors of low‐grade malignancy contained tumor‐cell subsets with reduced expression of one or several of the antigens. Eighteen lymphomas of high‐grade and 12 lymphomas of low‐grade malignancy contained varying tumor‐cell subsets that were negative for HLA‐DR, ‐DP, ‐DQ and li in a selective or combined manner. Three highly malignant tumors were devoid of all class‐II antigens and li; 2 highly malignant tumors expressed invariant chain only. The presence of high‐grade malignancy was significantly correlated with the occurrence of tumor cells lacking HLA‐DR (p = 0.004), HLA‐DP (p = 0.013), HLA‐DQ (p = 0.007) or 11 (p = 0.024).
Interferon treatment of human genital papillomavirus infection: Importance of viral type
Authors: Schneider Achim , Papendick Uwe , Gissmann Lutz , De Villiers Ethel‐Michele
Published: February 2007

Eighteen women and 6 of their male sexual partners with lower genital tract infections caused by various human papil‐iomavirus (HPV) types were treated with systemic or topical interferon (IFN) application. All patients with vulvar or penile lesions had a history of podophyllin or surgical treatment. In the female group, 9 patients showed complete response, 8 patients partial response and 1 patient no response. In the male patients complete response was seen in all patients. The response rate appears to depend on the HPV type present. Women with an HPV 16/18 infection showed a lower response rate to IFN treatment (complete response in 5 out of 14 patients), whereas lesions caused by HPV 6(11) showed com‐ plete response in all 4 cases. During follow‐up examinations (mean 7.5 months) no recurrence of disease was observed. Systemic treatment showed tolerable and temporary side‐ effects. Topical treatment yielded identical efficacy and no side‐effects.
Patterns of seminoma tissue markers and deletions
Authors: Yamamoto H. , Rudén U. , Ljungdahl‐Ståhle E. , Brehmer‐Andersson E. , Hirano K. , Hisazumi H. , Stigbrand T. , Wahren B.
Published: February 2007

Seminomas and control tissues were analyzed for several tumor markers. Very high levels of placental alkaline phosphatase (PLAP)‐like enzyme levels were found in all 18 seminomas studied. The majority of the seminomas were of phenotype I, thus differing from placental FLAP. The mean amount of enzyme protein as measured by monoclonal antibodies, was 100 times higher than in non‐malignant tissues and 10 times lower than in placental tissue. The specific enzymatic activity in seminomas was about half of that observed in placenta. Similarly, the specific activity of PLAP‐like enzymes in sera of patients with seminoma was only about half of that found in pregnancy sera. HCG was strongly elevated in 3 seminomas, but not obviously related to FLAP. Thirteen of the 17 pure seminomas had HCG over 100 IU/g, which was not seen in normal testes. Liver alkaline phosphatase (LAP) and intestinal alkaline phosphatase (IAP) were high in seminomatous tissues, the mean increases being 60‐fold and 20‐fold, respectively. The highest IAP levels were found in 2 yolk‐sac tumors. Ferritin was moderately elevated in seminomas, but high in several control tissues. Carcinoembryonic antigen (CEA) was not elevated and alpha‐fetoprotein (AFP) was not detected at all in pure seminomas. A decrease in carbohydrate antigen 50 (CA‐50) content was noted in seminomas as compared to normal testes, yolk‐sac tumors and choriocarcinomas. Defects in tumor‐related enzymes may account for increase of PLAP and decrease of CA‐SO.
Altered HLA antigens expressed on T and B lymphocytes of adult T‐cell leukemia/lymphoma patients and their relatives
Authors: Sonoda Shunro , Yashiki Shinji , Takahashi Kazuo , Arima Naomichi , Daitoku Yasuhisa , Matsumoto Makoto , Matsumoto Tadashi , Tara Mitsutoshi , Shinmyozu Koichi , Sato Kaoru , Inoko Hidetoshi , Ando Asako , Tsiji Kimiyoshi
Published: February 2007

The HLA types of peripheral blood lymphocytes (PBL) of 36 adult T‐cell leukemia/lymphoma (ATLL) patients were examined and compared with those of 45 healthy relatives of these patients, and with those of 10 non‐ATLL families including 80 healthy members. Thirty‐one percent of ATLL patients showed either a gain or a loss of HLA antigens determined by the presence of alien HLA antigens or the absence of inherent HLA antigens deduced from familial haplotype analysis. The antigen specificity of HLA gained or lost was variable and differed from case to case among ATLL patients. Although the gain of HLA was detected only in ATLL patients, the loss of HLA was found both in ATLL patients and in asymptomatic healthy relatives. The rate of HLA loss in ATLL patients (8.4%) and their relatives (17.8%) was much higher than in relatives of non‐ATLL patients (1.1 %). The HLA gain and loss revealed in the PBL of ATLL patients were confirmed by altered HLA phenotypes in the cloned T and B cells established from ATLL patients. Our results suggest that latently infecting HTLV‐1 may induce altered HLA phenotypes in T and B cells, primarily with loss of HLA antigens in a population of asymptomatic virus carriers, and secondarily with a gain of HLA antigens after the development of ATLL.
Alternative methods of selecting rat hepatocellular noduli resistant to 2‐acetylaminofluorene
Authors: Semple‐Roberts Esther , Hayes M. Anthony , Armstrong Dianna , Becker Richard A. , Racz William J. , Farser Emmanuel
Published: February 2007

Dietary 2‐acetylaminofluorene (2‐AAF) coupled with a stimulus for cell proliferation such as a 2/3 partial hepatectomy (PH) or a necrotizing dose of carbon tetrachloride is frequently employed to generate nodules of resistant (“initiated”) rat hepatocytes. This regimen is a useful model for experimental analysis of alterations in hepatocytes during carcinogenesis, and also as an assay for initiation by various carcinogens. Because of the decreasing availability of carcinogen‐containing diets from commercial sources, we have developed alternative methods of 2‐AAF administration to generate nodules in rats initiated with N‐nitrosodiethylamine. This study compared the nodule‐selecting and cancer‐promoting efficacy of 2‐AAF administered by the Solt‐Farber procedure (0.02% in diet for 2 weeks) with 2‐AAF administered by gavage, as a suspension in 1% aqueous carboxymethyl‐ cellulose (CMC). Three or 4 daily administrations of 2‐AAF by gavage (20 mg/kg/day) followed by PH on day 4 were equivalent to the dietary regimen in generating early resistant nodules, late persistent nodules and hepatocellular carcinomas. These regimens were similar to the dietary regimen of 2‐AAF in inhibiting virtually all normal hepatocyte proliferation. These regimens permit control over the duration and level of 2‐AAF exposure and the resulting size of selected nodules.
Effects of ouabain on NIH/3T3 cells transformed with retrovirai oncogenes and on human tumor cell lines
Authors: Tagliaferri Pierosandro , Yanagihara Kazuyoshi , Ciardiello Fortunate , Talbot Neil , Flatow Ursula , Benade Leonard , Bassin Robert H.
Published: February 2007

Both murine and human cell lines transformed by the v‐Ki‐ras gene have been shown to be much more sensitive to the toxic effects of the cardiac glycoside ouabain than their respective controls. This differential toxicity has previously been used in the isolation of flat revertant clones from populations of Kirsten murine sarcoma virus transformed NIH/3T3 cells. Here, we have undertaken a further characterization of this phenomenon in murine and human tumor cells. Two different techniques, a 51Cr‐release assay and a quantitative Crystal violet elution assay, have been employed to compare the sensitivities to ouabain of normal and v‐Ki‐ras‐transformed NIH/3T3 cells. In each assay, ras‐transformed NIH/3T3 cell lines displayed an increased sensitivity to ouabain as compared to the parental NIH/3T3 cell line, both in dose‐response and in time‐course experiments. In a separate study, ouabain was also able to inhibit the growth in semi‐solid medium of 2 v‐Ki‐ras‐transformed NIH/3T3 cell lines (DT and K‐NIH) in a dose‐dependent fashion. The same concentrations of ouabain were effective in both the 51Cr‐release and Crystal violet assays. To address the question of whether increased sensitivity to ouabain is a specific result of transformation with the ras oncogene or is a common event which accompanies transformation by other oncogenes, we have screened a variety of transformed NIH/3T3 derivatives. All of these lines displayed an increased sensitivity to ouabain when compared to the parental NIH/3T3 cell line.
Interferon‐induced alterations in metastatic capacity, class‐1 antigen expression and natural killer cell sensitivity of melanoma cells
Authors: McMillan T. J. , Rao J. , Everett C. A. , Hart I. R.
Published: February 2007

Pre‐treatment of B16 melanoma cells with recombinant interferon‐γ (IFN‐γ) markedly increased their lung‐colonising capacity following i.v. injection into syngeneic mice as compared with control cells. A similar enhancement was observed following the injection of treated cells into athymic nude mice but not in athymic mice carrying the beige mutation. Pre‐treatment of syngeneic mice with anti‐asialo GM1 antibody effectively abrogated any interferon‐induced increase in experimental metastatic activity. The same IFN‐γ treatment significantly increased resistance of B16 cells to splenic natural killer (NK) cell activity as determined by in vitro assays. IFN‐α/β pre‐treatment of B16 cells decreased sensitivity to NK‐cell‐mediated lysis to a lesser extent than IFN‐γ and had no detectable effect upon the subsequent metastatic activity of the tumour cells. Class‐1 antigen expression was altered by these IFN treatments, with IFN‐γ causing dramatic increases in expression of H‐2Db antigen, in a pattern consistent with the possibility that increased H‐2 antigen expression on B16 cells led to decreased NK‐cell sensitivity which was reflected by an increase in experimental metastatic capacity.
Metastatic but not primary melanoma cell lines grow in vitro independently of exogenous growth factors
Authors: Rodeck Ulrich , Herlyn Meenhard , Menssen Hans D. , Furlanetto Richard W. , Koprowski Hilary
Published: February 2007

Five out of 6 cell lines derived from metastatic melanoma lesions grew in a chemically defined base medium consisting of a mixture of calcium‐supplemented MCDB 153 and L 15 media in the absence of any polypeptide growth factors. In contrast, under these conditions no growth was seen in any of 5 primary melanoma cell lines tested, including 2 cell lines from patients whose metastatic cells proliferated well in base medium. Growth stimulation of all 11 melanoma cell lines by epidermal growth factor (EGF), transferrin, insulin, and insulin‐like growth factor (IGF)‐I alone and in various combinations was studied. Insulin represented the strongest single growth factor for primary and metastatic melanoma cell lines. The metastatic cell lines remained growth‐responsive to EGF, insulin and transferrin and responded more vigorously to these exogenously provided mitogens than the primary cell lines. No synergistic or additive growth effects of insulin, transferrin, or EGF for primary and metastatic cell lines were observed. Cross‐linking studies with 125I‐IGF‐I demonstrate surface expression of the type‐1 IGF receptor on melanoma cells. Growth stimulation by insulin and IGF‐I was inhibited by adding to the culture medium a monoclonal antibody to the type‐1 IGF receptor. Our studies indicate that IGF‐I and Insulin are major growth factors for melanoma cells and act via the type‐1 IGF receptor.
Paired Epstein‐Barr virus (EBV)‐negative and EBV‐converted Burkitt lymphoma lines: Stimulatory capacity in allogeneic mixed lymphocyte cultures
Authors: Avila‐Cariño J. , Torsteinsdottir S. , Ehlin‐Henriksson B. , Lenoir G. , Klein G. , Klein E. , Masucci M. G.
Published: February 2007

Epstein‐Barr virus (EBV)‐negative Burkitt lymphoma lines (BLE‐) and their in vitro EBV‐converted sublines (BLEc), obtained by infection with the P3HR1 and B95‐8 strains of EBV, were compared for their capacity to induce T‐lymphocyte proliferation in allogeneic mixed lymphocyte cultures (MLC). Regardless of the virus strain used for conversion, the BLEc lines induced a considerably stronger primary MLC response than their EBV‐negative parentals. Only the BLEc lines were able to maintain T‐lymphocyte proliferation in repeated stimulations. The low proliferative response observed in cultures stimulated with BLE−cells was not due to the generation of suppressor cells or to the release of inhibitory factors. The increased stimulatory capacity of BLEc lines was unrelated to changes in expression of MHC class‐I and class‐II antigen, or of B‐cell activation markers, and was not due to the reactivation of EBV‐specific memory T cells, since lymphocytes from EBV‐seropositive and seronegative donors responded similarly. The results indicate that the capacity of BL cells to elicit cellular immune responses may be influenced by their EBV‐carrying status.
Complementation of anti‐CEA and anti‐TAG‐72 monoclonal antibodies in reactivity to human gastric adenocarcinomas
Authors: Ohuchi Noriaki , Simpson Jean F. , Colcher David , Schlom Jeffrey
Published: February 2007

Monoclonal antibody (MAb) B72.3 and MAb COL‐4 are reactive with the high‐molecular‐weight (Mr > 106) tumor‐as‐sociated glycoprotein (TAG)‐72, and the Mr 180,000 carcino‐ embryonic antigen (CEA), respectively. Antibody competition radioimmunoassays (RIAs) using 1251‐MAb B72.3 or 1251‐COL‐4 have demonstrated that each MAb also recognizes a distinct antigenic determinant. Solid‐phase RIAs using MAbs B72.3 and COL‐4, however, demonstrated similar reactivity for each MAb with gastric carcinomas versus normal gastric mucosa. Tissue sections from ail of 17 gastric adenocarcinomas also reacted with both MAb B72.3 and MAb COL‐4 when immunoperoxidase techniques were used. Double‐staining techniques using both MAbs on the same section were performed on formalin‐fixed, paraffin‐embedded gastric tissue sections using the combination of avidin‐biotin peroxidase complex and avidin‐biotin alkaline phosphatase complex immunohis‐tochemical methods. The double‐staining technique revealed that some carcinoma cells react with MAb B72.3, some react with MAb COL‐4, and others react with both MAbs. This technique has demonstrated that more carcinoma cells can be detected by both MAbs as compared to the number of stomach carcinoma cells shown to be reactive with either one or the other MAb. These studies thus define the rationale for the use of combinations of MAbs which recognize different tumor‐associated antigens as immunological adjuncts for detection and perhaps therapy of gastric carcinoma.
Inhibition of homotypic aggregation of a human Burkitt‐lymphoma cell line
Authors: Wolf Michael F. , Koerner Ulrike , Klumpp Britta , Schumacher Kurt
Published: February 2007

Thi molecules involved in homotypic aggregation of the human Burkitt‐lymphoma cell line Raji were investigated by inhibition of reaggregation with carbohydrates and glycoconjugates, by inhibition of glycosylation, and enzyme treatment of the cell surface. Complete inhibition of reaggregation was achieved with bovine submaxillary mucin. Asialomucin, on the other hand, was not effective in this assay. Another potent inhibitor of reaggregation was the ganglioside GMI. The common carbohydrate structure of these molecules is NeuNAc‐(gal)‐galNAc. Lactosamine, fucosyllactosamine, sialyllactosamine, complex mannose type, or Thomsen‐Friedenreich antigen sequences are not involved in aggregation. Neuraminidase and chloroquine also abolished agglutination of cells. The finding that mucin, but not asialomucin, inhibits the reaction, demonstrates the importance of sialic acid in this process. Homotypic aggregation was shown to be resistant to trypsin. Using the glycosylation inhibitor tunicamycin we show that N‐glycosidically linked carbohydrate chains are involved in aggregation. Swainsonine or castanospermine, which inhibit processing of terminal sialyllactosamines to the mannose core, did not interfere with the reaction supporting the results of the inhibition assay. The data presented suggest the involvement of 2 molecules in homotypic aggregation of human Burkitt‐lymphoma cells. One component is a lectin‐like molecule containing N‐linked carbohydrate chains. The other component carries the neuraminidase‐sensitive and trypsin‐resistant determinant NeuNAc‐(gal)‐galNAc and, therefore, appears to be a glycolipid. This proposed lectin‐carbohydrate interaction in homotypic aggregation is further supported by the frequently observed dependence of lectins on divalent cations as indicated by inhibition of aggregation with EDTA and EGTA.
Cytogenetic studies on human myeloma cell lines
Authors: Jernberg H. , Zech L. , Nilsson K.
Published: February 2007

Cell lines (U‐266, U‐1957, U‐1996 and U‐2030) established from 4 patients with multiple myeloma (MM) were analyzed cytogenetically. The cell lines represent different stages in B‐cell differentiation as evidenced by ultrastructural and functional characteristics. The karyotypic pattern in 3 newly established myeloma lines was studied after a few months in culture and compared to the old myeloma cell line U‐266, which was examined after 6, 7 and 8 years of continuous cultivation. Frequency of progressive numerical and structural aberrations during long‐term cultivation and their correlation with alterations in growth properties were addressed. We describe the presence of a high frequency of both numerical and structural chromosomal abnormalities in the cells of all 4 myeloma lines studied. Chromosomes often associated with structural abnormalities were 1,3,6,12 and 14. A 14q + marker chromosome was detected in 2 of the 4 cell lines. The breakpoints on the chromosomes participating in structural aberrations in myeloma exhibit some correlation to chromosome sites at or close to locations of mapped oncogenes. No translocations of c‐myc were found. These data were further supported by Southern blot analysis (unpublished data). The extent of numerical, but not structural, aberrations correlates with the differentiation stage of the myeloma lines in that the 2 mature lines U‐266 and U‐1957 were both near‐diploid. Multiple progressive chromosomal changes have emerged in U‐266 during a period of 8 years with development of independence of feeder cells and increased growth rate. However, capacity for production of complete Ig molecules has remained stable.
Transplacental induction of a specific mutation in fetal Ha‐ras and its critical role in post‐natal carcinogenesis
Authors: Yamasaki H. , Hollstein M. , Martel N. , Cabral J. R. P. , Galendo D. , Tomatis L.
Published: February 2007

Mouse skin tumors were produced after transpiacental initiation [with 7, 12‐dimethylbenz(a)anthracene], only when the skin was treated post‐natally with a tumor‐promoting agent (12‐O‐tetradecanoyl phorbol 13‐acetate). DNA analysis of tumors showed that all carcinomas analyzed contained a specific mutation (A to T trans version) at the 61st codon of c‐Ha‐ras. Fifty per cent of the papillomas analyzed also had this same mutation. The A to T trans version at the 61st codon of Ha ras was heterozygous in all positive papillomas and carcinomas No such mutation was found when benzo(a) pyrene was used as an initiating agent. These results indicate that fetal c‐Ha‐ras can be trans placentally activated through a specific point mutation by a carcinogen, but a cell harboring such a mutation may remain dormant until it encounters a tumor promoting stimulus.
Enhanced metastases of a mouse carcinoma after in vitro treatment with murine interferon gamma
Authors: Ramani Pijamila , Balkwill Frances R.
Published: February 2007

We have studied the influence of inter ferons (IFNs) on the metastatic potential of mouse colon adenocarcinoma, COLON 26, cells. Pre‐treatment of the cells in vitro for 24 hr with recombinant murine IFN‐γ (rMuIFN‐γ) significantly in‐creased the number of lung tumour nodules when cells were injected i.v. into immunocompetent BALB/c mice and BALB/ c nude mice. However when MulFN‐γ‐pre‐treated cells were injected into beige (NK‐deficient) nude mice or anti‐asialoGM I (as GMI)‐serum‐treated BALB/c mice (NK‐depleted) no enhancement of metastatic potential was seen. Pre‐treatment of COLON 26 cells with recombinant human IFN‐α A/D (Bgl I), an IFN with equal activity on human and mouse cells, did net significantly enhance their subsequent metastases in immunocompetent or immunodeficient mice. In fact, there was a small but significant decrease in the number of tumour nodules in the lungs of beige nude and asGM I ‐treated mice. The e effects of rMuIFN‐γ on COLON 26 cells did not appear to be related to an alteration in MHC expression. COLON 26 eel Is constitutively express H‐2D and H‐2K antigens and both IFNs had equal enhancing (approx. 2‐fold) activity on the expression of these antigens at the doses used in this experiment (103 U/ml). We conclude that pre‐treatment with rMuIFN‐γ renders COLON 26 cells resistant to in vivo NK‐cell lysis via a mechanism that does not involve changes in MHC expression.
TPA induction of Epstein‐Barr virus early antigens in Raji cells is blocked by selective protein kinase‐C inhibitors
Authors: Lazdins Janis , Zompetta Claudia , Grimaldi Settimio , Barile Giuseppe , Venanzoni Marco , Frati Luigi , Faggioni Alberto
Published: February 2007

We have shown that modulation of intracellular calcium in EBV latently infected cells could induce the expression of viral an :igens, and suggested that a protein kinase‐C (PKC) may play a major role in the EBV genome activation. We now report further investigations on the role of PKC using 2 selective enzymatic inhibitors [l‐(5‐lsoquinolinylsulfonyl)‐2‐meth‐ylpiperazine] (H‐7) and Staurosporine. We show that these inhibitors can abrogate the inductive effect of TPA or the combination of TPA plus n‐butyrate. The inhibitors have no effisct on induction by calcium ionophores or by viral super‐infection. In this context the effect of verapamil (a specific calcium channel blocker) and of several calmodulin antagonists was investigated. No inhibitory effect of these agents could be demonstrated on any of the induction systems examined. These observations strengthen the idea that in some instances cellular PKC plays a role in the expression of viral antigens; however, alternative regulatory mechanisms cannot be excluded.
Correlation patterns of cancer relative frequencies with some socioeconomic and demographic indicators in Brazil: An ecologic study
Authors: Franco Eduardo L. , Filho Nelson Campos , Villa Luisa L. , Torloni Humberto
Published: February 2007

Sex‐specific relative frequencies (RF) of oral, esophageal, stomach, colon, rectal, laryngeal, lung, female breast, cervical and penile cancers obtained from a government‐sponsored, nation‐wide data base of histopathological diagnoses were evaluated with respect to all possible inter‐site correlations and with 12 socioeconomic and demographic variables for 23 States in Brazil. Use of bivariate and multivariate methods detected a high positive intercorrelation among RFs of lung, laryngeal and colon cancers regardless of sex. RFs for these 3 sites were also positively correlated with many markers of State development and affluence. Cervical and penile cancers emerged as a distinct subset with respect to their correlation patterns. RFs for these neoplasms were highly (positively) correlated (r = 0.8606, p < 0.001) with each other and exhibited intense negative associations with many of the affluence markers and the former cancer sites. Bivariate correlations generally exhibited a better fit with female‐specific RFs than with data from males which was reflected in the number of strong correlations detected by sex.
Up‐ and down‐regulation of human lymphokine (IL‐2)‐activated killer cell induction by monocytes, depending on their functional state
Authors: Nil Akihiko , Sone Saburo , Utsugi Teruhiro , Yanagawa Hiroaki , Ogura Takeshi
Published: February 2007

Studies were made to determine whether freshly isolated monocytes from healthy donors could influence the induction of lymphokine (IL‐2)‐activated killer (LAK) activity. Highly purified lymphocytes ( > 99%) and monocytes (>90%) were isolated by counter‐flaw centrifugal elutriation from peripheral blood. Lymphocytes incubated for 4 days with IL‐2 showed significant LAK activity against natural killer (NK) cell‐resistant target (Daudi) cells, whereas monocytes treated for 4 days with IL‐2 and/or IFN‐γ were not cytotoxic. Under the experimental conditions used, addition of monocytes to the lymphocyte cultures resulted in significant augmentation of the LAK activity, depending on the density of monocytes added. In contrast, monocytes stimulated by lipopolysaccha‐ride (LPS) markedly suppressed LAK activity induced by IL‐2, depending on the dose of LPS added. Similar up‐ and down‐regulations of LAK cell induction by monocytes were observed with 4 lines of human lung cancer cells as targets for LAK activity. Although supernatants from untreated monocytes did not increase LAK induction, supernatants from LPS‐stimulated monocytes suppressed LAK induction. The regulatory role of monocytes could not be replaced by the addition of exogenous interleukin I (IL‐1) or tumor necrosis factor (TNF). Prostaglandin E did not seem to play a regulatory role, since addition of indomethacin did not affect the regulation of LAK cell induction by monocytes‐ These results clearly indicate that human monocytes may cause up‐ or down‐regulation of the expression of IL‐2 induced LAK activity, depending on their functional state.
Human papillomavirus DNA in normal, metaplastic, preneoplastic and neoplastic epithelia of the cervix uteri
Authors: Fuchs P. G. , Girardi F. , Pfister H.
Published: February 2007

Colposcopically directed cervical punch biopsies from 362 patients were screened by Southern blot hybridization for the presence of DNA of human papillomavirus (HPV) 6, 10, 11, 16, 18, 31 and 33. The biopsies represented original squamous epithelium, Epithelium of metaplastic origin, different stages of cervical intraepithelial neoplasia (CIN) and invasive carcinomas. HPV6/11, 16, 18 and 31 were detected in 2.9% to 13.7% of histologically normal epithelia. HPV6/11 prevailed in CIN I. HPV16 was clearly more abundant than other HPV types in high‐grade CIN and invasive cancers (50%–60%), compared with healthy epithelium. Restriction enzyme cleavage analysis of DNA from primary cancers and corresponding metastases proved the stable association of HPVI6 DNA with invasive tumor cells. Preliminary follow‐up studies of CIN II patients suggested that HPV16‐associated lesions are relatively more likely to persist or to progress. Taken together, the data support the notion of a higher oncogenic potential of HPV16.
Trends in breast cancer incidence and mortality in Denmark, 1943–1982
Authors: Ewertz Marianne , Carstensen Bendix
Published: February 2007

The trend in breast cancer incidence and mortality was examined using data from the Danish Cancer Registry and the national mortality statistics respectively. The study population comprised £5,870 incident cases and 31,817 deaths from breast cancer in Denmark between 1943 and 1982. The incidence rate remained almost constant up to around 1960, whereafter it rose steadily. Comparatively little change was observed in mortality. Possible explanations for the differing trends in incidence and mortality include under‐reporting of breast cancer from death certificates and improvement in survival. A multivariate statistical analysis showed that the increase in incidence was due mostly to a cohort effect, though calendar time may have had a slight effect as well.
Decrease in tumor‐cell attachment and in a 140‐kDa fibronectin receptor correlate with greater expression of multiple 34‐kDa surface proteins and cytoplasmic 54‐kDa components
Authors: Rieber Manuel , Castillo Miguel A. , Ribbbr Mary S. , Irwin Juan C. , Urbina Cecilia
Published: February 2007

B 16 melanoma cells attach to matrix‐bound fibronectin but fail to adhere to albumin‐coated surfaces supplemented with soluble fibronectin. Attachment to substratum h also decreased in the presence of an adhesion‐disrupting antibody, or when cells are seeded on substrates poorly adhesive for these cells, such as collagen gels. We have now investigated some of the more general adhesion‐related alterations that occur between flattened and poorly attached cells. Immune blots of octylglucoside extracts with the adhesion‐disrupting IgG revealed a 140‐kDa component in flattened cells, in contrast to the increased detection of a 54‐kDa species in a comparable assay with rounded cells. Surface iodination also showed a decreased external exposure of a 140‐kDa fibronectin binding species and an increased labelling in multiple 34‐kDa protein species, in cells with decreased attachment to substratum. Analysis of 35S‐methionine‐labelled cell aggregates cultured on collagen gels also revealed a decrease in the 140‐kDa region and a greater labelling of multiple 54‐kDa components, compared to the same cells flattened on fibronectin. A change in 54‐ and 34‐kDa species was also teen in matrix‐associated components of rounded cells that failed to attach with soluble fibronectin. Since the 34‐kDa species increase in poorly adherent cells is mainly detected by inclination, and the 54‐kDa species increase in the same celts is partly associated with the corresponding detergent‐insoluble matrices, we propose that these I novel proteins may relate to cell rounding, through a trans membrane modulation involving both surface membrane and cytoskeletal structures.
Resistance to growth of the moloney lymphoma YAC in semi‐syngeneic graft recipients
Authors: Ährlund‐Richthr Lars , Klein Eva
Published: February 2007

The growth of the Motoney‐virus‐induced lymphoma YAC was examined in its strain of origin, A, as well ax in 6 AF1 hybrids, ALF1, AB6F1, ACBF1, AABYF1, AACAF1, and AASWF1, “Hybrid resistance”, i.e., lower frequency of incidence of tumor takes compared to simultaneous A controls, was observed in all of the tested hybrids. The results with the H‐2‐congeneic hybrids AABYF1, AACAF1 and AASWF1, indicated that H‐2 heterozygosity in itself was sufficient to exhibit hybrid resistance against YAC grafts. The A strain and its H‐2 congeneic AABYF1, AACAF1, and AASWF1, hybrids all had similarly low activity in NK assays. Thus, the elevated resistance detected in these H‐2 congeneic hybrids did nat correlate with NK activity. The rejection capacity of the ALF1, AB6F1, and ACBF1, hybrids correlated to their relatively higher NK activity, when compared to the NK low AABYF1, AACAF1, and AASWF1, hybrids. However‐, consideration of quantitative differences in between the hybrids which expressed elevated NK activity (i.e., ALF1, AB6F1, and ACBF1,) shows that the rejection capacity did nat correlate with NK status. The hybrid with the strongest NK effect (ACBF1) was the least resistant to YAC growth (27% palpable tumors), and the hybrid with the weakest NK effect (ALF1) was the most resistant to YAC growth (7.2% palpable tumors).
Radioimmunotherapy of human colon carcinoma by 131I‐labelled monoclonal anti‐cea antibodies in a nude mouse model
Authors: Buchegger F. , Vacca A. , Carrel S. , Schreyer M. , Mach J.‐P.
Published: February 2007

A mixture of 3 MAbs directed against 3 different CEA epitopes was radiolabelled with 131I and used for the treatment of a human colon carcinoma transplanted s.c. into nude mice. Intact MAbs and F(ab′)2 fragments were mixed because it had been shown by auto radiography that these 2 antibody forms can penetrate into different areas of the tumor nodule. Ten days after transplantation of colon tumor T380 a single dose of 600 μCi of 131I MAbs was injected i.v. The tumor grafts were well established (as evidenced by exponential growth in untreated mice) and their size continued to increase up to 6 days after radiofabelled antibody injection. Tumor shrinking was then observed lasting for 4–12 weeks. In a control group injected with 600 μCJ of 131I coupled to irrelevant monoclonal IgG, tumor growth was delayed, but no regression was observed. Tumors of mice injected with the corresponding amount of unlabelled antibodies grew like those of untreated mice. Bated on measurements of the effective whole‐body half‐life of injected 131I, the mean radiation dose received by the animals was calculated to be 382 rads for the antibody group and 478 rads for the normal IgG controls. The genetically immunodeficient animals exhibited no increase in mortality, and only limited bone‐marrow toxicity was observed. Direct measurement of radioactivity in mice dissected 1, 3 and 7 days after 131I‐MAb injection showed that 35, 7.2 and 7–2% of injected dose were recovered per gram of tumor, the mean radiation dose delivered to the tumor being thus more than 5,000 rads. These experiments show that therapeutic doses of radioactivity can be selectively directed to human colon carcinoma by i.v. injection of 131I‐labelled anti‐CEA MAbs.
Adhesion characteristics of human melanoma cell lines of varying metastatic potential
Authors: Ormerod E. Jane , Everett Christine A. , Hart Ian R.
Published: February 2007

The adhesive behaviour of a series of human melanoma cell lines, of varying metastatic potential, to basement membrane and stromal components was investigated in vitro. Experimental metastatic propensity was assessed from the number of pulmonary nodules formed after i.v. injection of cells into BALB/c nude mice. All cell lines showed similar kinetics of attachment when tested on plastic, type‐I collagen films, type‐I collagen hydrated gels, fibronectin, laminin type‐IV collagen substrates and bovine aortic endothelial monolayers. Fibro‐nectin‐coated plastic compared to plastic alone produced increased cell attachment and spreading to the same extent in all the cell lines. The melanoma lines attached preferentially to cryostat sections of lung compared to other organs reflecting the pattern of organ involvement of metastasis in vivo. However, no significant quantitative differences in attachment to lung sections were seen between melanoma variants of differing metastatic capacities. Cells labelled with [125I]iodo‐deoxyuridine to determine their initial organ distribution following i.v. injection showed that tumour‐cell arrest was not significantly changed enough to explain the differing metastatic capacities. Thus it appears that adhesive properties of these melanoma cells are not correlated with their capacity to form metastases in vivo.