bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterization and activation of cyclic adenosine 3′:5′‐monophosphate‐dependent protein kinase in C1300 murine neuroblastoma clones growing in vivo
Authors: Nakajima Fumiaki , Imashuku Shinsaku , Green Alexander A.
Published: February 2007

Cyclic adenosine 3′:5′‐monophosphate (cAMP)‐dependent protein kinases from three clones of C1300 neuroblastoma, established as growing tumors in A/jax mice, were identified and characterized. Mean (± SD) intratumor concentrations of cAMP ranged from 5.0±2.4 to 6.8±1.9 pmol/mg protein; cytosolic protein kinase activity ratios, calculated from the ‐cAMP/+cAMP value, ranged from 0.12±0.02 to 0.18±0.02. The total amount of cytosolic cAMP‐binding activity was highest in the NBA2 clone and nearly equal in N‐18 and NBP2. By DEAE chromatography, two peaks of cAMP‐binding activity eluting at ionic strengths of 0.09 and 0.15 M of NaCl were resolved from each clone. The latter peak was associated with catalytic activity (type II cAMP‐dependent protein kinase), whereas the former was not (free type I cAMP‐binding protein). Photoaffinity labeling with 8‐azido (N3)‐[32P]cAMP, followed by sodium‐dodecyl‐sulfate:polyacrylamide gel electrophoresis, resolved four major cAMP‐binding proteins with molecular weights ranging from 39,000 to 56,000 in each clone. The mol. wt 47,000 protein was judged to be a free regulatory subunit of type I kinase, and the mol. wt 39,000 protein a cleavage product. The mol. wt 54,000 and 56,000 proteins were both endogenously phosphorylated and both had a lower affinity for 8‐N3‐[32P]cAMP, characteristic of type II kinase. Selected properties of the cAMP‐binding proteins (kinetic and autophosphorylating features, response to tryptic hydrolysis, specificity for cAMP and temperature sensitivity) did not differ appreciably among clones. Despite initially low intratumor concentrations of cAMP, it was possible to activate the protein kinase system by simultaneous injection of N6, O‐dibutyryl‐cAMP and papaverine. This indicates that mouse C1300 neuroblastoma can be used profitably to study cAMP‐induced neuroblast differentiation.
Production of a mouse monoclonal antibody reactive with a human nuclear antigen associated with cell proliferation
Authors: Gerdes Johannes , Schwab Ulrich , Lemke Hilmar , Stein Harald
Published: February 2007

The production of a mouse monoclonal antibody, Ki‐67, is described. The Ki‐67 antibody recognized a nuclear antigen present in proliferating cells, but absent in resting cells. Immunostainings with Ki‐67 revealed nuclear reactivity in cells of germinal centres of cortical follicles, cortical thymocytes, neck cells of gastrointestinal mucosa, un‐differentiated spermatogonia and cells of a number of human cell lines. The Ki‐67 antibody did not react with cells known to be in a resting stage, such as lymphocytes, monocytes, parietal cells and Paneth's cells of gastrointestinal mucosa, hepatocytes, renal cells, mature sperm cells, brain cells, etc. Expression of the antigen recognized by Ki‐67 could be induced in peripheral blood lymphocytes after stimulation with phytohaemagglutinin, whereas it disappeared from HL‐60 cells stimulated with phorbol esters to differentiate into mature macrophages in a resting stage. These findings suggest that Ki‐67 is directed against a nuclear antigen associated with cell proliferation. A first series of immunostainings of tumour biopsies indicated that Ki‐67 may be a potent tool for easy and quick evaluation of the proportion of proliferating cells in a tumour.
Epstein‐barr virus (EBV)‐induced lymphoproliferative disease in cotton‐topped marmosets
Authors: Johnson Donald R. , Wolfe Lauren G. , Levan Göran , Klein George , Ernberg Ingemar , Åaman Pierre
Published: February 2007

Six cotton‐topped marmoset monkeys (Sangiunus oedipus) were inoculated with 105 tranforming units of B95‐8 virus, and two of them developed fatal lymphoproliferative disease. The EBV‐carrying tumor cells from these marmosets had the following characteristics: (I) they were polyclonal by surface immunoglobulin and immunoglobulin production in vitro; (2) they had no specific chromosome abnormalities, and (3) they failed to form colonies in large percentages in agarose. It is proposed that a spectrum of phenotypes of EBV‐induced lympho‐proliferative diseases in the cotton‐topped marmosets may be identified and are more akin to fatal infectious mononucleosis or X‐linked lymphoproliferative syndrome than to Burkitt's lymphoma.
Immunogenic variants obtained by mutagenesis of mouse mastocytoma P815. VI. Occasional escape from host rejection due to antigen‐loss secondary variants
Authors: Maryanski J. L. , Marchand M. , Uyttenhove C. , Boon T.
Published: February 2007

After mutagenesis of mouse mastocytoma P815, it is possible to obtain variant (tum−) clones that are almost always rejected by syngeneic DBA/2 mice. Most tum− clones express new variant‐specific antigens that can be detected by cytolytic T cells (CTL). Occasionally, mice injected with tum− variants eventually develop progressive tumors. Cells from these tumors were analyzed for antigen expression with variant‐specific and P815‐specific CTL clones. Out of 13 tumors examined, 11 were composed of cells that had clearly lost a variant‐specific antigenic determinant. These results indicate that tum− variants induce a specific host rejection response which usually results in complete elimination of the variant cells, but that occasional antigen‐loss constitutes an important mechanism of escape. The loss of a variant‐specific antigenic determinant from one tum− clone that had escaped rejection allowed the detection of a residual variant‐specific determinant. This was demonstrated by isolating a new CTL clone that lysed both the original tum− clone and its antigen‐loss variant, but not other P815 targets. Thus, some complex transplantation antigens can be separated into independent determinants by using antigen‐loss variants.
Persistent in vitro infection by human T‐cell leukemia‐lymphoma virus (HTLV) of normal human T‐lymphocytes from blood relatives of patients with HTLV‐associated mature T‐cell neoplasms
Authors: Ruscetti Francis W. , Robert‐Guroff Marjorie , Ceccherini‐Nelli Luca , Minowada Jun , Popovic Mikulas , Gallo Robert C.
Published: February 2007

Human T‐cell lymphoma‐leukemia virus (HTLV), a novel type‐C retrovirus, has been identified in tumor cells of patients with mature T‐cell neoplasms. Attempts to transmit the first isolate of HTLV designated HTLV‐I(CR) to secondary cultures of animal cells and several attempts to transmit HTLV to normal human T cells, activated with PHA and grown with T‐cell growth factor (TCGF), obtained from randomly selected normal adult donors were unsuccessful. Peripheral blood leukocytes were obtained from blood relatives of patients with HTLV‐positive T‐cell neoplasms. These normal adult T cells were exposed to HTLV, and in 5 of 10 samples tested from three separate families, persistent infection of cultured T‐lymphocytes by HTLV occurred. HTLV infection was monitored by three methods: an indirect membrane immunofluorescence assay for HTLV p19; an in situ hybridization assay for intracellular viral RNA; and electron microscopic analysis for extracellular viral particles. Two of the five infected T‐cell lines have been continuously positive for HTLV for over 24 months in continuous culture. B‐cell lines from the same individuals could not be infected by HTLV‐I(CR). Despite the release of viral particles from the infected cultures detected by electron microscopy, no extracellular reverse transcriptase was found. HTLV infection stimulated increased long‐term growth of the T cells although all HTLV‐I(CR)‐infected cultured cells always remained dependent on the addition of TCGF for growth. Phenotypic characterization of the infected cells by monoclonal antibodies, HLA typing, cytochemical and cell surface markers showed that all the infected cells were mature T cells and were easily distinguished from the cells used as a source of HTLV for infection. The infected cells were either of a predominantly helper or of a suppressor phenotype showing that both mature T‐cell subsets can be infected. Whether these data indicate that there is a genetic basis for susceptibility of human T cells to HTLV infection or reflect an increased sensitivity of some cells to infection by low titers of HTLV remains to be determined. These results provide further evidence for a predominant T‐cell tropism of HTLV.
Monocytic origin of the human hematopoietic cell line U‐937 and its convertibility to macrophages evidenced by isoenzyme mapping
Authors: Radzun Heinz J. , Parwaresch Mohammad R. , Sundström Christer , Nilsson Kenneth , Eissner Michael
Published: February 2007

U‐937 represents a well established permanent human hematopoietic cell line. Electron microscopical and enzyme cytochemical studies as well as the analysis of surface glycoproteins have provided ample evidence for the monocytic origin of U‐937. Upon stimulation with the tumour promotor 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA), U‐937 cells evolve into macrophage‐like cells with phagocytic capacities. Since human blood monocytes (BM) are characterized by five acid esterase (AcE; EC 3.1.1.6) isoenzymes which are cell‐specific in terms of isoelectric points (pi) and antigenicity, attempts were made in the present study to identify identical isoenzyme patterns in non‐ and TPA‐stimulated U‐937 cells. BM, cultured BM, non‐ and TPA‐stimulated U‐937 cells, as well as samples of resident human peritoneal macrophages (PM) as clearly‐defined functional derivatives of BM, were subjected to isoenzyme analysis using isoelectric focusing (IEF). The five monocyte specific isoenzymes of AcE were identified in both populations of U‐937. TPA‐stimulated samples showed two additional bands, identical to those appearing in cultured BM after 4 days of glass‐adherence and characteristic of resident human PM. Antisera raised against AcE of BM immunoprecipitated the two additional isoenzymes of TPA‐stimulated U‐937. It is concluded (1) the isoenzyme mapping of AcE documents the monocytic origin of U‐937. (2) TPA‐stimulation caricatures transformation of BM into resident tissue macrophages as far as pure morphology and AcE isoenzyme patterns are concerned. Thus, AcE isoenzyme mapping is apt for establishing reproducible and standardized criteria of different activation/differentiation states within the monocyte‐macrophage lineage.
Monoclonal antibody UJ 127:11 detects a 220,000‐240,000 kdal. Glycoprotein present on a sub‐set of neuroectodermally derived cells
Authors: Kemshead J. T. , Fritschy J. , Garson J. A. , Allan P. , Coakham H. , Brown S. , Asser U.
Published: February 2007

The monoclonal antibody UJ 127‐11 was raised following immunization of mice with human foetal brain and subsequent somatic cell hybridization of spleen cells with the mouse myeloma cell line P3‐X63‐Ag8‐653. Studies on normal foetal and adult tissues show that, by indirect immunofluorescence, the antigen recognized by UJ 127:11 is restricted in its expression to cells of neural rather than glial origin. Neural tumours such as neuroblastoma, medulloblastoma and ganglioglioma (neural component) bind the monoclonal antibody whereas malignancies originating from glial cells do not bind UJ 127:11. Biochemically the monoclonal antibody has been shown to bind to a glycoprotein of 220,000‐240,000 mol. wt. under reducing and non‐reducing conditions. Despite similarities in the molecular weight between human fibronectin and the antigen recognized by UJ 127:11, they have different serological and biochemical characteristics, suggesting that the monoclonal antibody is not binding to either cell or plasma fibronectin.
Clinical significance of circulating immune complexes in patients with metastatic breast cancer
Authors: Krieger G. , Kehl A. , Wander H.‐E. , Salo A.‐M. , Rauschecker H. F. , Nagel G. A.
Published: February 2007

Serum circulating immune complexes (CIC) were repeatedly measured by means of the Cl binding assay (Cba) and the Raji cell assay (Rca) in 158 patients with metastatic breast cancer (mbc). Frequency of occurrence and levels of CIC were only slightly increased in mbc when compared to age‐matched healthy women. They were identical to those of patients with localized breast cancer prior to mastectomy and to those of post mastectomy patients without evidence of recurrent disease. In mbc the results of the Cba and the Rca showed a poor correlation, whereas in a control group of patients with rheumatoid arthritis both tests showed significantly elevated levels of CIC. Patients with mbc were followed up clinically and biochemically by serially measuring CIC for an average of 10 months. Patients with positive CIC did not prove to be an unfavorable group regarding progression of disease and response to chemotherapy. When CEA and CIC levels were compared, CIC, unlike CEA, was a poor tumor marker. In conclusion, CIC as measured by Cl binding assay and Raji cell assay are not clinically significant tumor markers or prognostic indicators in patients with metastatic breast cancer.
Production of colony‐stimulating factors by Hodgkin cell lines
Authors: Burrichter Heinz , Heit Wolfgang , Schaadt Michael , Kirchner Hartmut , Diehl Volker
Published: February 2007

This work was performed with L 428, a cell line established in 1978 from a patient with Hodgkin's disease. These in vitro cells represent counterparts of in vivo Hodgkin and Sternberg‐Reed cells. L 428 and its derived sublines produce a significant amount of colony‐stimulatling factor (CSF) compared to standard preparations of CSF (fetal liver conditioned medium). Hodgkin‐cell‐derived conditioned medium, tested in cord‐blood assays and in semi‐solid agar systems, induced myeloproliferation predominantly. Production of CSF is independent of fetal calf serum concentration in the cultures. L 428‐conditioined medium could provide an excellent source for characterization and purification of granulocyte CSF.
Evidence for involvement of the immunoglobulin heavy‐chain gene locus in the 8:14 translocation of human B lymphomas
Authors: McIntosh R. V. , Cohen B. B. , Steel C. M. , Read Helene , Moxley Marion , Evans H. J.
Published: February 2007

Abnormalities of immunoglobulin (Ig) heavy chain synthesis occurring in human B lymphoid cell lines of malignant origin suggest that the Ig heavy‐chain gene complex is involved in the 8:14 chromosome translocation which these cells carry.
Changes in chromatin composition associated with hormone‐dependent mammary tumor regression
Authors: Goya Rodolfo G. , Sosa Yolanda E.
Published: February 2007

The chemical composition of chromatin from dimethylbenz(a)anthracene (DMBA)‐induced mammary tumors was compared in growing and regressing neoplasms. Tumor regression was induced by 2‐bromo‐α‐ergocryptine (CB‐154) administration to the tumor‐bearing rats. Seventy percent of the neoplasms showed a sharp fall in the levels of 3H‐thymidine incorporation into DNA after 7‐8 days of treatment (I mg CB‐154/day). These CB‐154‐responsive tumors showed a significant increase in non‐histone chromosomal proteins (NHCP) relative to DNA, while the histone fraction remained constant. Non‐responsive tumors showed no change in their chromatin composition. Low doses of ovine prolactin (18 IU) given together with CB‐154 (I mg) were able to prevent the effects of the latter on both DNA synthesis and chromatin composition of DMBA‐ induced mammary tumors. High doses of hormone (40 IU) produced a two‐fold increase in the tumors' DNA synthesis irrespective of the simultaneous administration of 1‐ or 2‐mg doses of CB‐154. This high dose of hormone was also able to prevent the effects of CB‐154 on chromatin composition of the tumors. We conclude that the above effects of CB‐154 are exerted mainly, if not exclusively, through the inhibition of prolactin secretion. On the other hand, the regression of these neoplasms after prolactin deprivation seems to be associated with an increase in the NHCP/DNA ratio. The possibility of an involvement of tumor cell membrane in the mechanism of regression of prolactin‐dependent mammary tumors is discussed.
Induction of epstein‐barr virus early antigens by corticosteroids: Inhibition by TPA and retinoic acid
Authors: Bauer G.
Published: February 2007

Corticosteroids can induce the synthesis of EBV antigens in the Burkitt lymphoma line Daudi. As early as 12 h after application of the drug, an increase of EA‐positive cells can be seen, the maximum induction being reached after 2 days. Nanogram amounts per ml of hormone are sufficient for measurable effects. Early antigen induction by corticosteroids does not require replication of viral DNA. Induction by corticosteroid differs from induction by other systems in two major respects: (1) it does not cooperate with other inducers, and (2) it is specifically inhibited by 12‐O‐tetradecanoyl‐phorbol‐13‐acetate (TPA). Induction by corticosteroids, however, shares at least one retinoic acid‐sensitive step with induction by chemicals such as TPA, 5‐iodo‐2‐deoxyuridine (IdUrd), n‐butyric acid (n‐BA) or inducing serum factor. This study defines three qualitatively different effects of TPA in Daudi cells: an inhibitory effect on EBV induction by corticosteroids and two differential types of synergistic effects with serum factor or n‐BA, respectively. In this particular cell line, TPA exhibits no inducing capacity when applied alone.
Progression of rat mammary development with age and its relationship to carcinogenesis by a chemical carcinogen
Authors: Sinha D. K. , Pazik J. E. , Dao T. L.
Published: February 2007

The progression of mammary gland development in Sprague‐Dawley rats between 5 and 440 days of age was studied. Tumorigenesis of the mammary gland after administration of 7,12‐dimethylbenz(a)anthracene (DMBA) was also investigated during different prepubertal and postpubertal stages. The mammary gland starts as a single primary duct on day 5 and with age branches and grows into a complete gland. The growth of the gland was observed to start from the nipple area and progress into the empty fat pad. This growth was in the form of club‐shaped end‐buds which, after the onset of puberty, became gradually transformed into alveolar buds. [3H]‐thy‐midine labelling index (LI.) was high (19%) in prepubertal glands, but it eventually declined to 8% at 60‐65 days. The postpubertal gland at the age of 150 days was determined to be practically mitotically static, but at 440 days, there was another increase in LI. There was a direct relationship between mammary tumorigenesis by DMBA and LI. in the mammary gland at the time of treatment with the carcinogen. Up to 55 days of age, tumorigenesis was 90‐100%, and LI. was also high during this period. The carcinogen was practically ineffective at 150 days, when the mammary gland was non‐proliferative. Though the total tumor induction was high in the prepubertal rats, the majority of the tumors (up to 84%) were benign fibro‐adenomas, while almost all the tumors in postpubertal rats were adenocarcinomas. Thus, a relationship was observed between the presence of ovarian hormones and the induction of malignant tumors. It was concluded that cell proliferation index at the time of carcinogen treatment is important for tumor induction, but whether or not the tumor is malignant is determined by the presence or absence of ovarian hormones.
Cloned lines of SJL/J spleen cells with cytotoxic reactivity
Authors: Riccardi Carlo , Allavena Paola , Ortaldo John , Herberman Ronald B.
Published: February 2007

Cultures of spleen cells from normal SJL/J mice were initiated and maintained in the presence of lectin‐free interleukin‐2 (IL‐2). After about 8 weeks in culture, the frequency of progenitors of proliferative cells became quite high (1/8‐1/10) and it was possible to obtain many clones by the limiting dilution technique. Some of the clones had high levels of cytotoxic reactivity against tumor target cells, even at low effectontarget ratios, indicating that natural effector cells from this low‐NK‐reactive strain have the capability of expressing potent lytic effects. There was considerable heterogeneity among the clones in their patterns of cytotoxic activity, morphology and expression of T‐cell‐associated antigens. Although the overall pattern of reactivity resembled that of NK cells, some clones reacted against an anchorage‐dependent lung cancer cell line, M109, without detectable effects against the other target cells tested, including the highly NK‐susceptible lymphoma cell line, YAC‐I. The pattern of cytotoxic reactivity by the clones did not correlate with the morphological appearance of large granular lymphocytes (LGL), which has been highly associated with NK activity of fresh, uncultured lymphoid cells.
Infection and transformation of fresh human umbilical cord blood cells by multiple sources of human T‐cell leukemia‐lymphoma virus (HTLV)
Authors: Markham Phillip D. , Salahuddin Syed Z. , Kalyanaraman V. S. , Popovic Mikulas , Sarin Prem , Gallo Robert C.
Published: February 2007

Human T‐cell leukemia‐lymphoma virus (HTLV) was first isolated from sporadic patients with adult T‐cell malignancies in the United States and subsequently from T‐lymphocytes established in culture from additional T‐cell leukemia‐lymphoma patients living in different geographical areas of the world. Co‐cultivation of normal unbilical cord blood with lethally irradiated, HTLV‐positive lymphocytes established in culture from many of these patients resulted in the productive infection of the cord blood T‐lymphocytes which grew in suspension culture in the absence of exogenous TCGF. These transformed cord blood cells have morphological and cytochemical properties similar to HTLV‐positive fresh and cultured tumor T‐cells and are distinguishable from virus donor cells by HLA haplotype and chromosomal markers. These cells express HTLV proteins, release type‐C virus particles and contain surface receptors for TCGF. These results demonstrate that HTLV isolated from T‐cell leukemic donors from different parts of the world can productively infect and transform fresh human cord blood T‐lymphocytes, and that the transformed cells share many similarities with fresh or cultured leukemic cells.
Resistance of fibroblasts and hematopoietic cells to ecotropic murine leukemia virus infection; an Akvr‐1R gene effect
Authors: Rasheed Suraiya , Gardner Murray B.
Published: February 2007

A gene named Akvr‐1segregates as a dominant allele in California wild mice (LC). Unlike Fv‐1 and Fv‐2 restriction alleles, the Akvr‐1confers resistance to replication of all ecotropic (N‐, B‐, and NB‐tropic) murine leukemia viruses (MuLV‐E) and prevents endogenous AKR virus‐induced lymphoma/leukemia in F1 hybrids of AKR × LC. We have tested fibroblasts and hematopoietic cells from mice of defined Akvr‐1 genotypes for in vitro susceptibility to various MuLV infections. Our results indicated a dominant in vitro resistance of exogenously MuLV‐E infected cells carrying Akvr‐1R allele although resistance was stronger in the homozygous cells than in the heterozygous cells. Moreover, resistance of cells to virus replication was not abrogated by high multiplicities of infection or by growth stimulation of hematopoietic cultures by various T‐ or B‐cell‐responsive mitogenic agents.
Expression of the bla antigen, defined by the monoclonal 38.13 antibody, on burkitt lymphoma lines, lymphoblastoid cell lines, their hybrids and other b‐cell lymphomas and leukemias
Authors: Klein George , Manneborg‐Sandlund Agneta , Ehlin‐Henriksson Barbro , Godal Tore , Wiels Joëlle , Tursz Thomas
Published: February 2007

The BLA expression of eight Burkitt lymphoma lines was high, whereas it was negative in four, including the two IgG producers tested. Most lymphoblastoid cell lines (LCL) of normal origin had only a low percentage of positive cells, not significantly above background, although a few had up to 30% positives. EBV conversion of the EBV‐negative Burkitt lymphoma line Ramos destabilized the high BLA expression, leading to a decrease in the average number of positive cells in the majority of the converted sublines in parallel with considerable fluctuation in antigen expression within each subline. Our group has previously shown that EBV‐conversion of Ramos cells can induce certain differentiation steps (Spira et al., 1981 a). EBV‐converted sublines of another EBV‐negative Burkitt lymphoma, BJAB, showed a much greater stability previously and remained unchanged with regard to BLA expression in our present experiments. Eight T‐cell leukemias, three myeloid leukemia lines and two diffuse histiocytic lymphomas were negative for BLA, whereas two myeloma lines were 30‐40% positive. A histiocytic tumor had marginal reactions. Hybrids derived from the fusion of high with low BLA‐reactive parental lines showed all three possible patterns (high, intermediate and low), provided that B‐cell lines were fused with each other. Fusion of two Burkitt lymphoma lines with the K562 erythroleukemia line led to the extinction of BLA expression, as well as to the eclipse of other B‐cell markers. B‐lymphoma and leukemia (CLL) cells harvested directly from the patient showed a heterogeneous reactivity pattern. Strong to intermediate BLA expression was found among CLL cells and in most histological groups of B‐CLL lymphomas except the centroblastic group (3/3 negatives). IgG‐expressing follicular lymphomas were less reactive than IgM ± IgD lymphomas of the same group. Immunocytomas were also low‐reactive. BLA can be thus expressed on a variety of B‐cell neoplasms; the degree of its expression appears to be related to the stage of differentiation.
Melanoma‐associated antigen p97 continues to be expressed after prolonged exposure of cells to specific antibody
Authors: Hellström Ingegerd , Brown Joseph P. , Hellström Karl Erik
Published: February 2007

p97, a 97,000 molecular weight cell surface glycoprotein associated with human melanoma, has attracted attention as a possible target for tumor therapy. Melanoma cells were grown overnight in the presence of IgG2a or IgGI monoclonal antibodies specific for p97, and then exposed to complement and also tested for binding of monoclonal antibodies specific for other epitopes of p97. The results indicated that melanoma cells continued to express p97 after prolonged exposure to specific antibody, a prerequisite for many therapeutic applications.
Serological responses of melanoma patients to vaccines derived from allogeneic cultured melanoma cells
Authors: Livingston Philip O. , Takeyama Hideo , Pollack Marilyn S. , Houghton Alan N. , Albino Anthony , Pinsky Carl M. , Oettgen Herbert F. , Old Lloyd J.
Published: February 2007

Twenty melanoma patients were immunized with a whole‐cell vaccine prepared from irradiated cells of the melanoma cell line SK‐MEL‐13. This line, derived from the melanoma of patient AH, expresses a differentiation antigen (initially defined by autologous antibody) which is restricted to melanomas and other cells of neural crest origin. The patients' sera were tested in resetting assays before and after vaccination for antibodies against cell surface antigens of autologous cultured melanoma cells and SK‐MEL‐13, and the specificity of observed reactions was defined by absorption tests. Nine patients developed antibodies against autologous cultured melanoma cells. In only one case, patient DM, were these antibodies directed against the AH antigen. In the remaining patients, the antibodies were related to fetal bovine serum in the vaccine growth medium. All patients developed antibodies against cell surface HLA alloantigens of the immunizing cell, SK‐MEL‐13, as demonstrated by absorption analysis with B cells from the donor of SK‐MEL‐13 and by serological typing with a panel of HLA typed peripheral blood T cells and B‐cell CLL cells. No specificity other than the AH antibody found in patient DM was detected after removal of alloantigens with B cells. We conclude that vaccines prepared from irradiated allogeneic melanoma cells expressing AH antigen, a demonstrably immunogenic differentiation antigen of the melanocyte lineage, are not effective in inducing AH antibody.
Cytogenetic studies on an epithelial cell line derived from poorly differentiated nasopharyngeal carcinoma
Authors: Sizhong Zhang , Xiukung Gao , Yi Zeng
Published: February 2007

An epithelial cell line, CNE‐2, has been recently established from a poorly differentiated nasopharyngeal carcinoma, and it represents the first of its kind. Using chromosome banding techniques, cytogenetic analysis of the cell line was carried out. It was demonstrated that the chromosome numbers of the CNE‐2 cells varied from 87 to 107 and the modal number was 104‐103. All cells contained a series of structurally abnormal chromosomes, and most of them were either consistent or frequently found. Among these chromosomes there were two giant markers which, by banding pattern analysis, proved to be distinct from the so‐called giant group A marker chromosomes previously found in many lymphoblastoid cell lines from NPC. Comparison between the CNE‐2 and CNE, another epithelial cell line, which was established from well‐differentiated squamous NPC, showed that while they were quite different in many cytogenetic aspects, they had three marker chromosomes in common, namely, an iso8q, a t(?;3q) and a small acrocentric one. The question of whether chromosome markers specific for NPC exist is discussed in the light of the data presented.
Detection of epstein‐barr virus IgA/EA antibody for diagnosis of nasopharyngeal carcinoma by immunoautoradiography
Authors: Zeng J. , Gong C. H. , Jan M. G. , Fun Z. , Zhang L. G. , Li H. Y.
Published: February 2007

An immunoautoradiographic method was used for the detection of EB virus lgA/EA antibody in sera from NPC patients and other control groups. Ninety‐six percent of NPC patients had IgA/EA antibody with a high titer of GMT. The positive rates of IgA/EA antibody in patients with malignant tumours other than NPC and in normal individuals were only 4% and 0%, respectively. Eleven patients histologically diagnosed as having a chronic inflammation and who showed positive for IgA/EA antibody by immunoautoradiography were rebiopsied; six of them were discovered to have squamous cell carcinoma. Fourteen NPC patients had no IgA/EA antibody detected by immunofluorescence and immunoenzymatic testing, but 11 and six of them had IgA/VCA and IgA/EA antibodies detected by immunoautoradiography, respectively. These data indicate that the immunoautoradiographic method is more sensitive than either the immunofluorescence or immunoenzymatic test for the detection of IgA/EA antibody, and can be used for the detection of NPC in the early stages of development.
3H‐estradiol and 3H‐R5020 binding in cytosols of normal and neoplastic human ovarian tissue
Authors: Pollow K. , Schmidt‐Matthiesen A. , Hoffmann G. , Schweikhart G. , Kreienberg R. , Manz B. , Grill H.‐J.
Published: February 2007

High‐affinity cytoplasmic estrogen and progesterone receptors in normal and abnormal ovarian tissues were studied. Estradiol receptor was detectable in 65% and progesterone receptor in 36% of the malignant tumors; 39% of all malignant ovarian tissues were estradiol‐ as well as progesterone‐receptor‐positive. Tumors were said to be receptor‐positive when the receptors bound >5 fM steroid/mg cytosol protein. No correlations were found between receptor status and histopathological diagnosis. In normal ovarian tissues collected at various phases of the menstrual cycle no changes in [<3>H]‐estradiol and [<3>H]‐R5020 binding could be detected. Analysis of the receptor concentration for both steroid hormones with regard to the menopausal status demonstrated highest levels in postmenopausal women. No significant difference could be found when two groups of patients with advanced ovarian carcinoma associated with the cytosol estrogen receptor status were compared in terms of two different therapeutic schemes (cytosolic chemotherapy with and without tamoxifen).
H‐2 control of the cytotoxic antibody response against a newly defined MuLV‐related cell‐surface antigen: G(B10.A)
Authors: Vlug Arjen , Zijlstra Maarten , DeGoede Ruud E. Y. , Hesselink Wim G. , Schoenmakers Harrie J. , Melief Cornelis J. M.
Published: February 2007

H‐2‐congeneic C57BL mice with milk transmission of B‐tropic murine leukemia virus (V+ mice) have a much higher lymphoma incidence than the same strains without milk‐transmitted virus (V—mice). Gene(s) within the major histocompatibility complex (H‐2) influence virus titers, lymphoma incidence, lymphoma type and the anti‐MuLV envelope antibody response. In this paper, we report that the prevalence of cytotoxic antibodies to virus‐induced lymphomas is also regulated by the H‐2 complex. Milk transmission of MuLV resulted in the formation of cytotoxic antibodies against primary virus‐induced C57BL lymphomas. These antibodies detect an antigen that is also present on the RADAI tumor‐cell line, and on normal spleen cells of young adult B10.A (H‐2a) mice of both V+ and V‐ sublines, but not on spleen cells of young adult B10 (H‐2b) mice of either subline. These cytotoxic antibodies were detected in the sera of B10V+ and B10.A(5R)V+ animals, but not in the sera of B10.AV+ mice. This indicates that the prevalence of these antibodies is controlled by a gene in the K‐and/or I‐A region of the H‐2 complex. The presence of these cytotoxic antibodies in serum is recessively inherited. The specificity of the cytotoxic antibodies was investigated with a standard panel of transplantable tumor‐cell lines. Of these, only the RADAI cells expressed the target antigen in direct cytotoxicity tests and by absorption. The ability of B10V+ sera to lyse the B10.AV+ and RADAI tumor cells is ascribed to antibody activity against a new MuLV‐related cell‐surface protein: G(B10.A). Immunochemical analysis and absorption experiments with different types of purified MuLV and MuLV‐infected cell lines indicate that the cytotoxic antibodies belong to low‐avidity IgM antibodies that are directed to MuLV.
Characterization of the surface proteins of SV40‐transformed mouse and human cells: Absence of SV40‐specific proteins
Authors: Rose Timothy M. , Weil Roger
Published: February 2007

The proteins of a number of SV40‐ and spontaneously transformed mouse and human cell lines were compared in an effort to identify a surface protein which would correspond to the SV40 tumor‐specific transplantation antigen (TSTA). Analysis of the one‐ and two‐dimensional electrophoretic patterns of 35S‐methionine‐labelled total proteins and 125I‐labelled surface proteins of several of these cell lines failed to reveal the presence of proteins specific to transformation by SV40. Antisera were prepared against SV40‐ and spontaneously transformed mouse cells in syngeneic mice. In serological assays, these antisera reacted with surface antigens common to both SV40‐ and spontaneously transformed mouse cell lines. Electrophoretic analysis of the 125I‐surface‐labelled proteins which these antisera immunoprecipitated from extracts of SV40‐ and spontaneously transformed mouse and human cells identified a set of common surface proteins with apparent molecular weights of 15, 44, 50, 72, 77, 105, 150 and 230 kdal. No SV40‐specific surface proteins were detected. Two of the transformed cell surface proteins (105 and 150kdal) were present as well in membrane fractions of 35S‐methionine‐labelled primary mouse kidney cultures. The proteins of the primary cultures could not be iodinated by lactoperoxidase suggesting that these proteins were present at a “cryptic” location at the surface of normal cells. We were not able to obtain serological or immunochemical evidence for the presence of SV40 large T‐antigen at the surface of any of the SV40‐trans‐formed cell lines tested using either hamster anti‐SV40 tumor sera, a rabbit antiserum against SDS‐denatured gel‐purified large T‐antigen or antisera against SV40‐transformed mouse cells. In conjunction with the report that large T‐antigen released from disrupted SV40‐transformed cells will bind to cell surfaces (Lange‐Mutschler and Henning, 1982), we consider the possibility that the specific rejection of SV40‐induced tumors by sensitized animals is the result of immunological reactions against both common transformation‐related surface antigens and SV40 T‐antigen from disrupted cells that has bound to the surface of other tumor cells.
Antitumor effects of interferon in mice injected with interferon‐sensitive and interferon‐resistant friend leukemia cells. III. Inhibition of growth and necrosis of tumors implanted subcutaneously
Authors: Belardelli F. , Gresser I. , Maury C. , Duvillard P. , Prade M. , Maunoury M.‐T.
Published: February 2007

Administration of highly purified interferon to DA/2 mice inhibited the growth of interferon‐sensitive or interferon‐resistant Friend erythroleukemia cells implanted subcutaneously. Injection of interferon at the site of tumor inoculation was more effective than injection of interferon intraperitoneally. Histologic examination of tumors in interferon‐treated mice showed extensive areas of tumor‐cell necrosis in the absence of an obvious host‐cell infiltrate. Interferon inhibited the growth of established subcutaneous tumors and induced complete tumor regression in some mice. Interferon treatment also resulted in an inhibition of tumor metastases in the liver and spleen.
Characterization of “H‐2K,D‐like structures” on MM2 × mouse L cell hybrids. II. An “H‐2Kd‐like” structure on C3H‐derived tumor
Authors: Kubota Koichi , Manson Lionel A.
Published: February 2007

Anti‐MM2 serum, which had been prepared by immunizing C3H/He mice with syngeneic MM2 mouse mammary ascites tumor, immunoprecipitated “H‐2K,D‐like” molecules on DBA/2 and C3H.H‐2° lymph node cells as well as on somatic cell hybrids between MM2 tumor and mouse L cells. Preclearing of lysates from C3H.H‐2° lymph node cells with anti‐H‐2.31 serum removed all “H‐2K,D‐like” molecules reactive with anti‐MM2 serum, indicating that the molecules detected by anti‐MM2 serum are H‐2Kd antigens. The anti‐H‐2.31 serum detected an “H‐2K,D‐like structure” on the hybrid cells and absorption of the anti‐H‐2.31 serum with the hybrid cells deprived the serum of anti‐H‐2Kd reactivity. The hybid cells could induce antibodies against the H‐2Kd antigen in C3H/He mice. These results indicate that on the hybrid cells, whose parental cells were both derived from C3H mice, there is an “H‐2K,D‐like structure” that has the H‐2Kd private specificity. Absorption of anti‐MM2 serum with EL4 cells did not affect the capacity of the serum to detect the H‐2Kd antigen on C3H.H‐2° lymph node cells, indicating that the “H‐2Kd‐like structure” is distinct from “H‐2K,D‐like structure A” which was previously reported. Nine isozymes were examined and MM2 cells, mouse L cells, and the hybrids were found to have the same isozyme markers as those of the C3H/He mouse.
Emergence of low‐mobility t lymphocytes in cancer patients and its clinical significance
Authors: Nakajima Toshifusa , Mori Takeo , Iwaguchi Takao
Published: February 2007

Electrophoretic mobility of lymphocytes from healthy individuals and cancer patients was analyzed by means of a fully automated cell electrophoresis apparatus (Parmo‐quant). Electrophoretic mobility histograms showing two distinct peaks were obtained with peripheral blood lymphocytes from normal subjects. A border line was placed at the mobility of 0.95 μm/sec/V/cm, and cells slower than this were designated as low‐mobility cells (LMC) and faster cells as high‐mobility cells (HMC). LMC, constituting the slower peak, consisted mainly of B cells and a small number of T cells (LMT). HMC, the constituent of the faster peak consisted of two types of T cells (HMT). In patients with malignant tumors, analysis revealed lowered electrophoretic mobility of T cells, with T/B ratios comparable to those in normal subjects. An increase in the percentage of LMT seemed characteristic of suppressed immunity in cancer patients since it showed a strongly positive correlation with the percentage population of IgG‐FcR‐positive T lymphocytes and with the serum level of immunosuppressive substance. The incidence of abnormal LMT/HMT ratios was highest in patients with recurrent cancer, and it was also high in the advanced stages of gastrointestinal cancer. This index could provide useful information for the early detection of cancer relapse during postoperative follow‐up. A fully automated electrophoresis apparatus greatly facilitated the measurement (5 min/100 cells) and permitted quantitative and statistical analysis of mobility histograms of cell electrophoresis.
Excretion of a tumor‐associated trypsin inhibitor (TATI) in urine of patients with gynecological malignancy
Authors: Huhtala Marja‐Liisa , Kahanpää Kari , Seppää Markku , Halila Hannu , Stenman Ulf‐Håkan
Published: February 2007

In earlier studies we reported the finding of a tumor‐associated peptide that also occurred at high concentrations in early amniotic fluid. Determination of the N‐terminal sequence of this peptide revealed that it is closely related or identical to the pancreatic secretory trypsin inhibitor. Therefore, the peptide is called tumor‐associated trypsin inhibitor (TATI). The concentration of TATI was determined by radioimmunoassay in the urine of 148 patients with various forms of gynecologic malignancy and in a reference population consisting of 98 patients with non‐malignant gynecologic disease, and also in 40 patients with severe infections or inflammatory disease. In the reference population, the median urinary concentration of TATI was 22 μg/g creatinine and the central 95 % reference interval was 7‐50 μg/g creatinine. Elevated urinary levels were observed in 53 % of all patients with gynecologic cancer, in 63 % of those with active disease and 26% of those in clinical remission. The highest urinary TATI level (11,000 μg/g creatinine) was over 200 times the upper limit of the reference range. Patients with cervical cancer had the highest frequency of elevated values. Increased excretion of TATI was also observed in patients with severe bronchopulmonary infections and pancreatitis. Although increased excretion of TATI is not cancer‐specific, the distinction by elevated levels of TATI between malignant and nonmalignant gynecologic disease is better than by most other putative tumor markers, and the increased excretion of TATI in patients with active disease can be important for the understanding of tumor biology.
Site‐directed chemotherapy with a drug bound to anti‐idiotypic antibody to a lymphoma cell‐surface IgM
Authors: Hurwitz Esther , Kashi Rina , Burowsky Daphna , Arnon Ruth , Haimovich Joseph
Published: February 2007

Xenogeneic antibodies against the cell‐surface IgM of a B‐cell lymphoma (38C‐13) were coupled through a dextran bridge to the anti‐neoplastic drug, daunomycin. The conjugate maintained both its antibody and its drug activity. The effectiveness of the conjugate was tested in vivo in mice challenged with the 38C‐13 lymphoma. Drug conjugates of the idiotypic antibodies injected intraperitoneally 2 days after tumor transplantation almost completely inhibited tumor development. The controls, daunomycin‐dextran‐goat anti‐DNP, free daunomycin, or the antibodies alone had no effect or only delayed the tumor development.
Characterization of T lymphocytes mediating in vivo protection against RSV‐induced murine sarcomas
Authors: Prat Maria , Di Renzo Maria Flavia , Comoglio Paolo M.
Published: February 2007

Tumors induced by Rous sarcoma virus (RSV) in different mouse strains share a common tumor‐associated transplantation antigen, whose expression is controlled both by the viral transforming srcgene and by cellular gene(s) (Prat et al., 1981). The mechanism(s) responsible for the in vivo protective immune response against two RSV‐induced sarcomas has now been investigated. In the “Winn assay” the growth of both tumors was specifically prevented or significantly reduced by the simultaneous administration of lymphocytes isolated from immunized donors. The effector cells were radiosensitive (2,500 rad), Lyt‐l+, Lyt‐2,3+ Thy.l+ cells. Better protection was afforded by transfer of immune spleen cells from mice pre‐treated with cyclophosphamide, which is known to abrogate T‐cell suppressor activity. In a 5‐day mixed lymphocyte/tumor cell culture specific anti‐RSV‐induced sarcoma cytotoxic activity was barely detected, while a good production of interferon‐gamma (IFN‐y) was observed. The cells involved showed the same functional and surface phenotype as displayed by the effectors of the “Winn” assay. It is concluded that in the immune rejection of RSV‐induced sarcomas, Lyt‐l+, Lyt‐2,3+ T cells, rather than cytotoxic T lymphocytes, and lymphokines such as IFN‐γ are involved.
In vitro induction of tumor‐specific immunity by highly purified vsv grown in SV40‐transformed cells and tumor glycolipids incorporated into liposomes
Authors: Ansel Sandra
Published: February 2007

Cytotoxic effector lymphocytes (CL) were induced by in vitro immunization of spleen cells from normal Syrian hamsters to syngeneic tumor cells, either SV40‐transformed (EH‐SV) or spontaneously transformed (EH‐N). The lymphocyte reactivity was measured in a direct 15Cr release cytotoxic effector lymphocytes with EH‐SV‐ and EH‐N‐labelled targets. A specific cytotoxic effect against tumor cells carrying the sensitizing antigens was observed. Cytotoxic effector lymphocytes were also induced by in vitro immunization of hamster spleen cells to highly purified vesicular stomatitis virus (VSV) grown either in syngeneic SV40‐transformed fibroblasts or in “normal” fibroblats. Purified virus possessing an intact envelope or virus subparticles devoid of their glycoprotein spikes stimulated the cellular immune responses against host tumor antigens present within the viral envelope. Cytotoxicity assays have revealed two tumor‐specific antigens (TSA), one induced by SV40 and present in SV40‐transformed cell lines and the other present in “normal” cells. CL were also induced by in vitro sensitization of spleen cells from normal hamsters to liposomes containing the polar glycolipid fraction from EH‐SV and/or EH‐N cells. A specific cytotoxic effect against tumor cells that have supplied the glycolipid extract was observed, suggesting specific recognition of glycolipid antigens characteristic for each tumor line. This study supports the view that surface glycolipids act as tumor‐specific antigens implicated in the destruction of SV40‐induced tumors in Syrian hamsters.
A leukocyte adherence inhibition (LAI) assay of anti‐tumor immunity in rats using selective radioimmunological assessment of adherence of T lymphocytes and monocytes
Authors: Morizane Toshio , Sjögren Hans Olov
Published: February 2007

A new, highly sensitive micro‐glass‐tube leukocyte adherence inhibition (LAI) assay has been developed to detect anti‐tumor sensitization in a rat colon carcinoma system. This technique requires fewer cells and smaller amounts of antigen preparations than the previous glass tube method. The microscopic enumeration of adherent cells is replaced by a cellular radioimmunoassay (CRIA) which utilizes antibodies binding to mononuclear cells (MNC) and 125I‐labelled protein A. Antigen preparations were shown to be adsorbed to glass. Precoating of glass vials with fetal calf serum or antigen preparations was shown to cause a major increase in nonspecific LAI. The new LAI technique is designed to minimize such nonspecific LAI. By applying an anti‐T‐cell monoclonal antibody (McAb), an anti‐lg antiserum and an anti‐monocyte (MC) antiserum it became possible to assess selectively the change of adherence of T cells, B cells and monocytes. The specificity of the assay was demonstrated with anti‐T‐cell and anti‐MC reagents in criss‐cross experiments using two defined antigens and different tissue extracts as test antigens. The LAI response of peripheral blood MNC was examined after the subcutaneous isografting of 1 × 107 X‐irradiated DMH‐W49 colon carcinoma cells. Six days after sensitization all the rats showed an LAI response detectable with anti‐T‐cell and anti‐MC reagents, which gave considerably higher LAI indices than anti‐MNC antiserum. After the subcutaneous isografting of 1 × 105 viable DMH‐W49 cells, 12 of 24 rats (50%) showed an LAI response detectable with the anti‐MNC antiserum during a 3‐week follow‐up period. On the other hand, an LAI response was detected in all the 14 tumor‐inoculated rats tested when the adherence of T lymphocytes and monocytes was assessed selectively.