bluuebunny/crossref_metadata_2025 huggingface dataset:
In vitro reaction of cancer patients and others to Bacillus Calmette‐Guérin
Authors: T Gaye , Chan Chi Wing , Cochran Alistair J. , Kennedy Robin , MacKie Rona M. , Morrison Lindsay J. A.
Published: February 2007

We examined the in vitro reaction of leukocytes from 71 cancer patients and 121 control donors (disease‐free individuals or patients with non‐malignant non‐debilitating conditions) against Bacillus Calmette‐Guérin using a one‐stage capillary leukocyte migration assay. The proportion of reactive cancer patients and control donors increased with increasing BCG concentration, but the proportion of patients who reacted was less than that of control donors at all concentrations of BCG. As cancer stage advanced and the volume of clinically detectable tumor increased, the proportion of reactive patients decreased. There was a small age‐related decline in reactivity in the control population. The results of simultaneous in vitro tests and Mantoux reactions were concurrent in a majority of normal individuals so tested.
Benzo(a)pyrene metabolism and DNA adduct formation in serially cultivated strains of human epidermal keratinocytes
Authors: Parkinson E. Kenneth , Newbold Robert F.
Published: February 2007

Six strains of human epidermal keratinocytes were shown to be capable of metabolising benzo(a)pyrene (BP)for at least 40 population doublings in culture. Metabolism was independent of human or mouse fibroblast products, and also the growth rate of the epidermal cultures, at least when cholera toxin and epidermal growth factor were included in the medium. Epidermal strains metabolized and bound to their DNA more BP than human fibroblasts, and themselves exhibited a 2‐to 3‐fold range of inter‐strain variability. Chromatog‐raphic analysis of epidermal DNA containing bound BP showed that the major metabolite which had reacted with the DNA was a 7,8 dihydrodiol ‐ 9,10 oxide of BP (the proposed ultimate carcinogenic metabolite of BP).
Organization of actin‐containing cables in cultured skin fibroblasts from individuals at high risk of colon cancer
Authors: Kopelovich Levy , Lipkin Martin , Blattner William A. , Fraumeni Joseph F. , Lynch Henry T. , Pollack Robert E.
Published: February 2007

Actin‐containing cables were examined by immuno‐fluorescence in cultured skin fibroblasts from individuals genetically prone to colon cancer. The study confirmed our earlier finding of an altered distribution of actincontaining cables in skin fibroblasts of patients with hereditary adenomatosis of the colon and rectum (ACR) (Kopelovich et al., 1977). Abnormalities were also found in about one‐half of the asymptomatic offspring at risk for ACR, while a polyposis‐free branch of one ACR family showed a normal pattern of actin‐containing cables. Persons from colon cancer‐prone (CCP) families without polyposis, and normal controls, showed no disturbance in the actin patterns. The results suggest that this phenotypic marker may be useful in identifying ACR gene carriers and in probing cellular controls of carcinogenesis.
Possibly specific immune complexes in sera of patients with untreated acute myelogenous leukemia
Authors: Fäldt R. , Ankerst J.
Published: February 2007

Eleven patients with acute myelogenous leukemia (AML) in the acute stage of the disease were studied. Sera of seven patients were assayed for complement‐dependent cytotoxicity to autochthonous AML cells and sera of four patients were tested with allogeneic target cells before and after ultrafiltration at low pH. No specific cytotoxic activity could be detected in any untreated sera. Ultrafiltration at low pH led, however, to the appearance of a significant cytotoxic activity in all of seven autochthonous and in three of four allogeneic combinations. Identical treatment of control AB sera did not result in any similar activity. The results show that potentially cytotoxic antibodies to leukemia‐associated antigens are present in the acute stage of acute myelogenous leukemia. However, they do not express complement‐dependent cytotoxicity in untreated sera. After ultrafiltration at low pH significant complement‐dependent cytotoxicity was detectable in most of the sera. One possible explanation is that cytotoxic antibodies to leukemia‐associated antigens are present in the form of circulating immune complexes. Splitting of them at low pH and elimination of the antigen component might result in an increased amount of free cytotoxic antibodies.
Monoclonal antibody against a naturally occurring rat mammary carcinoma
Authors: Gunn Barbara , Embleton M. J. , Middle J. G. , Baldwin R. W.
Published: February 2007

Inbred rats were hyper‐immunised against a syngeneic transplanted mammary carcinoma, Sp4, which originally arose spontaneously in a breeding female. Spleen cells from Sp4‐immune rats were fused with P3‐NSl‐Ag 4 mouse myeloma cells, and supernat‐ants from the hybrid progeny were screened for antibody reacting with Sp4 cells using an isotopic antiglobu‐lin assay. One hybrid culture was reactive with Sp4 cells but not against other target cells, and this hybrid was cloned in soft agar. Many Sp4‐positive clones were isolated and one (Sp4/A4) was chosen for detailed study. Clone Sp4/A4 was positive only with Sp4 cells and failed to react with cells of other tumours or nomal tissues. It contained much higher titres of antibody than detected in immune sera, and it is suggested that this approach offers a practical method for producing highly active tumour‐specific antibodies.
The effect of age on the histopathogenesis of 7,12‐dimethylbenz(a)‐anthracene‐induced mammary tumors in the lewis rat
Authors: Haslam Sandra Z.
Published: February 2007

The influence of age at time of DMBA administration on (l) the histopathogenesis and site of mammary tumor origin and (2) the precancerous nature of DMBA‐induced mammary dysplasias was examined in Lewis rats between 25 and 200 days of age. Rats were killed at various times after carcinogen treatment and the numbers of tumors and dysplasias were recorded from microscopic examination of mammary gland wholemounts. We found that, in all age‐groups tested, mammary tumors always appeared to originate within mammary end‐buds and terminal ductules and that terminal due‐tule hyperplasia appeared to be an early stage in tumor formation. Age‐related differences in susceptibility to tumor development were paralleled by similar differences in susceptibility to the development of terminal ductule hyperplasias. In contrast, although age‐related differences in susceptibility to the development of hyperplastic alveolar nodules were also noted, they did not correspond to those observed for tumor development. Thus, hyperplastic alveolar nodules did not appear to be a site of tumor origin in situ or directly related to tumor development.
Multiplicity‐dependent biological and biochemical properties of epstein‐barr virus (EBV) rescued from non‐producer lines after superinfection with P3HR‐1 EBV
Authors: Cho Myung‐Sam , Fresen Karl‐Otto , Hausen Harald Zur
Published: February 2007

Superinfection of lymphoblastoid cells of EBV non‐producer lines with non‐transforming P3HR‐l EBV leads to the rescue of transforming virus. At least part of the recovered molecules represent recombinant DNA between superinfecting P3HR‐l EBV and resident viral genomes (Fresen et al., 1979, 1980). With high titer stocks of superinfecting P3HR‐l EBV, viral particles with early antigen (EA)‐inducing properties can be rescued, indicating that under these conditions of infection input viral genomes may become replicated. Sequential blot analysis with 32P‐P3HR‐l EBV DNA of intracellular viral DNA synthesized following infection reveals a multiplicity‐dependent pattern. High particle inputs lead to preferential synthesis of certain fragments, independent of infection of either EBV genome‐free or EBV genome carrier cells. This accumulation of specific viral sequences indicates defective replication of P3HR‐l EBV DNA.
Virus infections and recurrence of osteosarcoma in patients receiving human leukocyte interferon
Authors: Ingimarsson Snorri , Carlström Gun , Cantell Kari , Strander Hans
Published: February 2007

Human leukocyte interferon was given as adjuvant treatment for osteosarcoma in 26 patients. Blood from these patients was studied at regular intervals for antibodies to various micro‐organisms. A continuous record of clinically manifest infections during the course of interferon therapy was made in 23 of the 26 patients. Distant metastases arose in 14 patients ‐ the metastasis group. The report presents comparisons between these patients and the non‐metastasis group. Seroconversion in tests with any of the studied micro‐organisms was not common. Most of the seroconversions were not associated with clinical symptoms. Seroconversion during interferon therapy was confined to patients in the metastasis group. In some cases there was a chronologic link with early evidence of metastasis. The results of the serologic investigations are discussed and are correlated to the course of interferon therapy and of the neoplastic disease.
Reduction of suppressor cells in cancer patients treated with OK‐432 immunotherapy
Authors: Uchida Atsushi , Hoshino Takashi
Published: February 2007

Ok‐432, a streptococcal preparation, was intradermally injected daily into patients with advanced cancer of the stomach or lung for 4 weeks and the effects of OK‐432 on the mitogenic responses of cancer patients were followed. The cells involved in the depression of the response of untreated cancer patients were characterized. The cells responsible for impaired responses were nylon wool non‐adherent and suppressed the mitogen responses of autologous and allogeneic lymphocytes. These cells lost their suppressive activity during 7 days' culture in vitro. Following OK‐432 immunotherapy, mononuclear cells from cancer patients showed increased responses to PHA and Con A, and nylon wool non‐adherent cells did not inhibit the mitogen responses. These results suggest that the cells suppressing non‐specific mitogen responses are sensitive to in vitro culture, belong to nylon wool non‐adherent cells and are lost during 4 weeks of OK‐432 therapy.
Human tumor‐lymphocyte interaction in vitro. VII. Blastogenesis and generation of cytotoxicity against autologous tumor biopsy cells are inhibited by interferon
Authors: Vánky F. , Argov S.
Published: February 2007

In a high proportion of cases blood lymphocytes from cancer patients cultured with autologous tumor biopsy cells undergo blastogenesis. In addition, cytotoxicity against autologous tumor cells is generated. These auto‐activated lymphocytes also kill the highly NK‐sensitive K‐562 and Molt‐4 cells. Addition of interferon (IF) at the initiation of the cultures inhibited blastogenesis and the generation of autologous cytotoxicity. On the other hand, lymphocytes already activated for autologous killing in the mixed cultures were not affected by short‐term IF treatment prior to the cytotoxic assay. Lymphocytes cultured alone lose their cytotoxicity against K‐562. However, when IF was present in these cultures the lymphocytes were cytotoxic for K‐562. In addition, lymphocytes cultured for 6 days under conditions which did not result in activation killing of Molt‐4 cells could be induced by short‐term incubation of the effectors with IF.
Chromosomal alterations of rat cell lines transformed by human adenovirus type‐12 virion, whole DNA and left‐end DNA fragments
Authors: Yamashita Toshiharu , Yamamoto Kohtaro , Shiroki Kazuko , Kimura Genki , Matsui Katsuaki , Osato Toyoro , Shimojo Hiroto , Fujinaga Kei
Published: February 2007

A normal rat cell line, 3Y1‐B clone 1‐6 (3Y1) and its adenovirus (Ad) type‐12‐transformed derivatives, W4 (transformed by Ad12 Virion), WY3 (transformed by Ad12 whole DNA), CY1‐1 (transformed by the Ad12 EcoR1‐C fragment, left 16.5%), GY1‐1 (transformed by the Ad12 Hind11‐G fragment, left 6.8%) and HY1 (transformed by the Ad12 Acd‐H fragment, left 4.7%) were studied cytogenetically. 3Y1 and some of the transformed cell lines (W4, WY3 and GY1‐1) were diploid or pseudodiploid, while others (CY1‐1 and HY1) were hypotetraploid. A metacentric marker M1 was detected in GY1‐1 cells and another marker M2 in W4 and CY1‐1 cells at a high frequency. By the Giemsa banding technique, the metacentric markers M1 and M2 from these fully transformed cell lines were identified as iso‐chromosomes derived from 1q (M1) and 3q (M2), respectively. On the other hand, the markers were detected only at a low frequency in incompletely transformed HY1 cells. However, hypersomy in chromosome No. I was observed at a high frequency in this cell line. It can be concluded that hypersomy of chromosomes No. 1 or 3 found in transformants and metacentric markers found in complete transformants are characteristic features in rat cells transformed by Ad12.
Specificity and biological activity of extracted murine tumor‐specific transplantation antigens
Authors: Pellis Neal R. , Yamagishi Hisakazu , Kahan Barry D. , MacEka Catherine M.
Published: February 2007

Crude 3m KCI extracts of the chemically induced C3H/ HeJ fibrosarcomas, MCA‐F and MCA‐D, possess two immunobiologically active components upon fractionation by preparative isoelectric focusing. Strongly acidic (pl 2.5‐3.6) antigens induced potent tumor facilitation while mildly acidic (pl 5.8‐6.0) proteins engendered resistance to neoplastic cell challenge. The possibility that these proteins were tumor‐specific transplantation antigens (TSTA) was investigated in specificity tests using fractions prepared from the MCA‐F and MCA‐D. Pre‐treatment of individual groups of mice with the tumor‐facilitating and immunizing fractions from extracts of two antigenically different fibrosarcomas MCA‐F and MCA‐D followed 10 days later by challenge of all groups with both neoplasms revealed both responses to be specific for the tumor of origin. The immunizing fractions from MCA‐F and MCA‐D engendered 50% (p<0.01) and 60% (p<0.01) reduction in the growth of their respective tumor cell challenge. Facilitating antigens of both tumors specifically potentiated only the growth of their parent neoplasms by more than 75% (p<0.01). Non‐specific protection was not observed, demonstrating that both fractions bore TSTA activity. Both the resistant state and the condition of facilitated tumor growth engendered by the isolated tumor cell fractions could be adoptively transferred to normal syngeneic recipients via lymphoid cells, and the im‐munoprotective fraction evoked specific delayed‐type hypersensitivity in immunized hosts. Assessment of the activity of the partially purified TSTA revealed that the immunizing fraction of MCA‐F exhibited narrow range dose‐response properties. Immunoprotection was demonstrable within a restricted range whether the antigen was administered as whole cells, crude extract, or pIEF purified material. These results indicate that (1) separate tumor‐facilitating and immunizing activities are common features of carcinogen‐induced tumors; (2) the antigen‐induced facilitation of, and resistance to, tumor cell challenge is mediated by lymphoid cells; (3) both moieties display distinct antigenic specificity in vivo; and (4) the restricted range of immunoprotection is a property intrinsic to at least some carcinogen‐induced tumors and their antigenic subcellular fractions.
Alterations in the extracellular matrix organization associated with the reexpression of tumorigenicity in human cell hybrids
Authors: Der Channing J. , Stanbridge Eric J.
Published: February 2007

The expression of fibronectin on the cell surface was evaluated on a series of intraspecific human cell hybrids formed between HeLa and normal fibroblast strains. Although these hybrids continued to express many of the in vitro transformation properties of their corresponding tumorigenic HeLa parent, they were now unable to form tumors when inoculated into athymic nude mice. From these suppressed hybrid populations, rare tumorigenic segregant subpopulations arose which had regained their tumorigenic capacity. A comparison of the expression of fibronectin on the cell surface was made between these tumorigenic segregant cell lines and their corresponding non‐tumorigenic HeLa/fibroblast hybrid. Following specific immunofluorescent staining for fibronectin, a striking alteration in the cell surface organization was observed to correspond with the reexpression of tumorigenicity in these hybrids. Tumorigenic HeLa/fibroblast hybrids were also significantly altered in both their cellular and colonial morphology. Double immunofluorescent staining to simultaneously visualize both surface fibronectin and collagen revealed that these two extracellular matrix proteins displayed an extensive degree of codistribution and expressed a coordinate shift in organization which correlated with the appearance of tumorigenic segregant hybrid populations. These observations are in agreement with the apparently close structural association between fibronectin and collagen and suggest that the organization of these two components in the extracellular matrix may be an important determinant for in vivo growth potential.
Expression of alien minor histocompatibility antigens distinct from tumor‐specific transplantation antigen on a murine fibrosarcoma
Authors: Parmiani Giorgio , Colombo Mario , Ballinari Dario
Published: February 2007

The fibrosarcoma ST2, induced by 3‐methylcholan‐threne in BALB/c (H‐2d) mice, also expressed alien histocompatibility antigens of the C3Hf and B10 background not encoded by the MHC. To examine the relationship between these alien, minor antigens and the tumor‐specific transplantation antigen (TSTA) of the tumor, in vivo immunogenicity test were performed in BALB/c mice and in hybrids between BALB/c and C3Hf (H‐2k), C3H.OH (H‐202), C3H.SW (H‐2b), BALB.K (H‐2k), B10.BR (H‐2k), and B10.D2 (H‐2d) mice. A significant loss of TSTA immunogenicity was found in (BALB/c X C3Hf) and in (BALB/c X C3H.OH)F1 animals and, to a lesser extent, in (BALB/c X C3H.SW)F1 mice as compared to the immunogenicity of the tumor in BALB/c mice. Immunogenicity tests with ST2 in BALB/c X (BALB/c X C3Hf) or in BALB/c X (BALB/c X B10.D2) backcross mice, respectively, revealed that half of the BALB/c X (BALB/c X C3Hf) and 97% of the BALB/c X (BALB/c X B10.D2) animals were able to mount an immune response to ST2. To see whether the loss of TSTA immunogenicity in (BALB/c X C3Hf) was due to common determinants shared between TSTA and alien non‐H‐2 C3Hf antigens or to a genetically linked low responsiveness to TSTA introduced by C3Hf and C3H.OH strains, BALB/c mice were immunized with normal normal tissues of some BALB/c X (BALB/c X C3Hf) back‐cross, anti‐ST2 resistant mice. Normal tissues of anti‐ST2 resistant, dd and dk typed backcrosses were able to immunize BALB/c mice against a challenge of an otherwise lethal dose of ST2 cells. Some but not all BALB/c X (BALB/c X B10.D2) anti‐ST2 resistant donors had tissues able to immunize BALB/c hosts against the ST2 growth. Since resistance to tumor growth and expression of minor “alien” antigens shared with the tumor segregate independently, we concluded that alien, minor C3Hf and B10 antigens of the BALB/c sarcoma ST2 are distinct from the TSTA of this tumor.
Herpesvirus saimiri: Protective effect of attenuated strain against lymphoma induction
Authors: Wright John , Falk Lawrence A. , Wolfe Lauren G. , Deinhardt Friedrich
Published: February 2007

An attenuated strain of Herpesvirus saimiri (A‐HVS) induced a persistent, latent infection in common marmosets. Virus was recovered from circulating lymphocytes of infected animals and antibody response to virus‐specified antigens was demonstrated over 3‐12 months PI. A‐HVS‐infected common marmosets and animals without previous A‐HVS exposure were challenged with oncogenic HVS (125‐825 PFU); marmosets without A‐HVS infection developed malignant lymphoproliferative disease and died 3 weeks PI whereas all persistently‐infected animals have survived 38‐43 months post challenge.
The role of gp70 in the target antigen recognized by murine leukemia virus immune cytotoxic T‐lymphocytes
Authors: Watson Andrew , Bach Fritz H.
Published: February 2007

The role of the murine leukemia virus (MLV) envelope glycoprotein, gp70, in the formation of the target antigen recognized by in vivo generated MLV‐immune cytotoxic T‐lymphocytes (FMR‐MLV immune T) was studied. Treatment of the Rauscher MLV‐transformed C57BL/6 target cell, RBL‐5, for 4 h with the protein glycosylation inhibitor tunicamycin, was found to render the cells resistant to lysis by FMR‐MLV immune Tc while lysis of the tunicamycin‐treated RBL‐5 cells by allogeneic H‐2b immune Tc was not decreased. This failure of FMR‐MLV‐immune Tc to lyse tunicamycintreated RBL‐5 cells correlated with an inhibitory effect of tunicamycin on the synthesis of gp70, resulting from impaired glycosylation of the gp70 precursor, gpPr90. Yet, examination of the tunicamycin‐treated RBL‐5 cell surface by lactoperoxidase 125I iodination and immunoprecipitation techniques, revealed (I) that neither unglycosylated gp70 nor gpPr90 antigens were detectable and (2) that the gp70 antigens from tunicamycin‐treated and untreated RBL‐5 cells were identical in their (I) migration pattern in SDS polyacrylamide gel electrophoresis (SDS PAGE) (2) isoelectric focusing profiles on a pH 3.5 ‐ pH 10 gradient and (3) external, lactoperoxidase accessible, 125I peptides. Further, tunicamycin‐treated RBL‐5 cells retained 33% ‐ 50% of their Rauscher MLV gp70 expression, as determined by quantitative absorption of a goat anti‐Rauscher MLV gp70 serum. When this latter result was interpreted with reference to the study by Lesley et al. (1974), who observed a linear relationship between the amount of alloantigen expressed and susceptibility to immune Tc killing, it was concluded that this quantitative change in gp70 expression could not adequately account for the failure of FMR‐MLV immune Tc to lyse tunicamycin‐treated RBL‐5 cells. It is therefore proposed that an MLV‐induced antigen other than gp70 may be recognized on RBL‐5 cells by FMR‐MLV immune Tc.
Destruction of erythroleukemia, myelocytic leukemia and burkitt lymphoma cells by photoactivated protoporphyrin
Authors: Malik Zvi , Djaldetti Meir
Published: February 2007

The effect of protoporphyrin on erythroid, myeloid and lymphoid leukemic cells and their destruction induced by the photoactivated porphyrin was studied. Friend erythroleukemic cells (FL) and myelocytic leukemic cells (ML) accumulated protoporphyrin in a cap or patch‐like pattern observed by fluorescence microscopy. Photoactivated protoporphyrin induced the appearance of “holes” on the cell membrane demonstrated by scanning electron microscopy. On the other hand, Burkitt lymphoma (BL) and mastocytoma (MS) cells accumulated porphyrin intracellularly around the nuclear envelope and as circular profiles, respectively. Photoactivated protoporphyrin induced development of multiple blebs on the cell membrane, and even complete cell destruction. Cytotoxicity of protoporphyrin at short‐term incubation periods was determined by [3H]thymidine and [3H]uridine incorporation. Protoporphyrin, unexposed to light, reduced the incorporation of both precursors only to a moderate extent. On the other hand, porphyrin‐treated cells exposed to light showed complete inhibition of RNA and DNA synthesis. Long‐term exposure of ML and BL cells to porphyrin in the dark induced a nearly 50 % inhibition of RNA and DNA synthesis. Although the cytotoxic effect of protoporphyrin in the dark was lower than that of photoactivated porphyrin, this may possess a potential activity in vivo even without illumination.
Colony morphology, cell growth and viral radiosensitivity as parameters for detecting strain differences in epstein‐barr virus (EBV)
Authors: Levitt Mark , Shungu Daniel , Henderson Earl
Published: February 2007

Several parameters of two lymphocyte‐transforming Epstein‐Barr viruses (EBVs), B95‐8 and Olare, derived from infectious mononucleosis (IM) and African Bur‐kitt's lymphoma (BL), respectively, were compared to determine if there were biological differences between Epstein‐Barr viral strains. Morphology of transformed colonies, growth of transformed cell lines, sensitivity to UV light and X‐irradiation, as well as EBV‐related, antigens were investigated in autochthonous cell lines. The morphology of transformed colonies was found to differ consistently both in new transformants and in lines which had been continuously growing in culture for almost I year. The BL transformants grew to greater density than did the IM transformants, and BL‐derived virus was more sensitive to both UV light and X‐irradiation. No immunologic differences were noted in the strains in assays performed for EBV‐related antigens.
Establishment of a lymphoid cell line from leukemic cells of a patient with chronic lymphocytic leukemia
Authors: Karande A. , Fialkow P. J. , Nilsson K. , Povey S. , Klein G. , Najfeld V. , Penfold G.
Published: February 2007

Two lymphoid cell lines were established from a patient with chronic lymphocytic leukemia by infecting blood cells with Epstein‐Barr virus (EBV). Studies of morphology, glucose‐6‐phosphate dehydrogenase, malic enzyme, immunoglobulin, and chromosomes of the two lines indicated that one of them originated from leukemic cells while the other arose from residual normal blood cells. The morphology and capacity for immunoglobulin secretion in the line that arose from leukemic cells were similar to those found in EVB‐carrying lymphoblastoid cell lines grown from patients without neoplasia and differed from those seen in fresh chronic lymphocytic leukemia cells. These observations suggest that the introduction of EBV into the leukemic cells may have caused them to differentiate in a fashion similar to that noted in normal B cells after exposure to EBV.
Chemical transformation of cultured human skin fibroblasts derived from individuals with hereditary adenomatosis of the colon and rectum
Authors: Rhim Johng S. , Huebner Robert J. , Arnstein Paul , Kopelovich Levy
Published: February 2007

Chemical transformation of cultured human skin fibroblasts (PF) derived from individuals with hereditary adenomatosis of the colon and rectum is reported. Cells treated only with various levels of N‐methyl‐N′‐nitro‐N‐nitrosoguanidine (MNNG) underwent morphological alteration. The morphologically altered cells formed large aggregates when suspended in liquid growth medium above an agar base and grew to high saturation densities. One altered (MNNG, 1.0 μg/ml) cell culture formed colonies in soft agar. Transformed cells were resistant to rechallenge of MNNG (I μg/ml) and showed a more prolonged life‐span compared to the untreated cells. Altered cells became heteroploid cells. However, no progressively growing tumors were produced when cells were inoculated subcutaneously into nude mice. The data suggest that chemical carcinogens alone may not induce neoplastic transformation of fibroblasts from humans genetically predisposed to cancer and that neoplastic transformation of these skin cells by chemical carcinogens might require the presence of a tumor promotor and the use of an immuno‐privileged site in the nude mouse system.
Quantitation of methylglyoxal bis(guanylhydrazone) in blood plasma and leukemia cells of patients receiving the drug
Authors: Seppänen P. , Alhonen‐Hongisto L. , Siimes M. , Jänne J.
Published: February 2007

Methylglyoxal bis(guanylhydrazone), a cytostatic compound which apparently interferes with the metabolism and/or functions of the natural polyamines (spermidine and spermine), was effectively taken up by cultured human lymphocytic leukemia cells, rapidly resulting in the formation of a concentration gradient of up to 1,000‐fold across the cell membrane in cells grown in the presence of micromolar concentrations of the drug. For an anti‐proliferative effect on the leukemia cells, an intracellular concentration of more than 0.5 mm was required. The uptake of methylglyoxal bis‐(guanylhydrazone) was critically dependent on the growth rate of the leukemia cells. Low intracellular concentrations of the drug were present in cells growing slowly, whereas in rapidly dividing cells the intracellular concentration of the drug approached 5 mm. When given as repeated intravenous infusions to two leukemic children, methylglyoxal bis(guanylhydrazone) exhibited sharp and transient peaks of plasma concentration, the drug having an apparent half‐life in plasma of only 1‐2 h. However, as in cultured cells, the drug was rapidly concentrated in the leukemia cells, reaching concentrations that were distinctly anti‐proliferative. In contrast to the rapid disappearance of methylglyoxal bis(guanylhydrazone) from plasma, the circulation leukemia cells retained the drug for a period of several days with only minimal decrease in the initial concentrations. Methylglyoxal bis(guanylhydrazone) was given to the patients for 1 to 2 months as intravenous infusions, the timing of which was determined by regular assays of the drug concentrations in the leukemia cells. In agreement with the results obtained with the cultured cells, an intracellular concentration of about 0.5 to I mm was apparently required for growth‐inhibitory action to occur. Regular determination of the cellular drug concentrations indicated that methylglyoxal bis(guanylhydrazone) could be given as weekly infusions. This treatment schedule represents much lower dosing of the drug than the earlier daily regimens which were commonly associated with unacceptable toxicity.
Preparation and identification of human breast epithelial cells in culture
Authors: Easty G. C. , Easty D. M. , Monaghan P. , Ormerod M. G. , Neville A. M.
Published: February 2007

The routine preparation of gram quantities of lobuloalveolar and ductal structures from human reduction mammaplasties by treatment with collagenase is described. When cultured on plastic or glass surfaces these structures give rise, initially, to epithelial sheets consisting of cells which retain many morphological characteristics of myoepithelial cells and do not stain with an antiserum which reacts with the surfaces of the lining epithelium of breast ducts and lobulo‐alveoli. Subsequently, cells which resemble the lining epithelial cells migrate from these structures and react strongly with the antiserum. Cell proliferation in these cultures is minimal. Explants of major ducts dissected from mastectomies produce vigorously proliferating epithelial sheets which contain morphologically similar cells not identifiable as either lining epithelium or myoepithelium, although nearly all the cells in direct contact with the medium react with the lining epitheliumspecific antiserum.
Clonal origin and trisomy of chromosome 15 in murine B‐cell malignancies
Authors: Fialkow Philip J. , Reddy A. Lakshma , Bryant Jean I.
Published: February 2007

During the course of a multifaceted study of clonality in murine neoplasms we observed two B‐lymphoid malignancies. Results of studies with the X‐chromosome‐linked enzyme phosphoglycerate kinase strongly suggest that these tumors had a clonal origin. Each of them had trisomy 15. This chromosomal abnormality has been found consistently in many murine thymic neoplasms, and has been thought to be specific to tumors of thymic origin. However, the occurrence of trisomy 15 in each of the only two B‐cell malignancies thus far detected in our studies indicates that it may occur in Blymphoid progenitors as well.
The activation state of macrophage subpopulations from a murine fibrosarcoma
Authors: Moore K. , McBride W. H.
Published: February 2007

We have separated subpopulations of macrophages from an immunogenic fibrosarcoma by the technique of unit gravity velocity sedimentation. The activation state of these subpopulations was determined by measurement of the Fc receptor avidity of adherent cells, and by their 5′ nucleotidase and acid phosphatase activity. The subpopulations were compared to resident peritoneal macrophages and the peritoneal macrophage subpopulations elicited by injection of proteose peptone or C. parvum. The results show that two macrophage subpopulations exist within the tumour. The smaller, peroxidase‐positive population, with a sedimentation velocity of 1‐5 mm/h, is similar to proteose peptonestimulated macrophages with respect to Fc receptor expression, while the other, rapidly sedimenting population (5‐9 mm/h) is partially activated. However, neither population achieved the level of activation demonstrated by rapidly sedimenting, C. parvum‐activated macrophages. Analysis of the enzyme activity of rapidly adherent macrophages indicated that tumour, proteose peptone or C. parvum macrophages were all activated when compared to resident peritoneal macrophages. No significant differences were found with respect to the elevated levels of acid phosphatase in the three activated macrophage populations, but the 5′ nucleotidase activity of C. parvum‐elicited macrophages was significantly lower than either the proteose peptone or tumour macrophages. This again demonstrated that the tumour macrophages were less activated than C. parvum macrophages. These data show that tumour‐infiltrating macrophages are a heterogeneous population composed of at least two subpopulations existing in different activation states and that within the tumour microenvironment they are not capable of differentiating to the higher activation state, demonstrated by C. parvum macrophages.
Metastatic tumor cells adhere preferentially to the extracellular matrix underlying vascular endothelial cells
Authors: Kramer Randall H. , Gonzalez Robert , Nicolson Garth L.
Published: February 2007

Two metastatic cell lines, mouse B16‐Fl melanoma and human Hs939 melanoma, were examined for their abilities to adhere to confluent vascular endothelial cell monolayers and to the underlying endothelial extracellular matrix. Tumor cells attached slowly to the endothelial cell monolayers while they adhered rapidly to isolated extracellular matrix. When analyzed by poly‐acrylamide gel electrophoresis in sodium dodecylsulfate solutions, the extracellular matrix was shown to be primarily composed of a protein of identical migration and molecular weight to fibronectin. Tumor‐cell adhesion to fibronectin‐coated polyvinyl surfaces mimicked the rapid rate of attachment of tumor cells to extracellular matrix, and tumor cells adherent to either extracellular matrix or fibronectin‐coated polyvinyl dishes adopted an unusual, highly spread and flattened morphology with numerous small projections. These results suggest that fibronectin associated with the endothelial basement membrane may be, in part, responsible for establishing an adhesive gradient that could be important in malignant cell extravasation.
Enhanced cytocidal action of methotrexate by conjugation to concanavalin A
Authors: Tsuruo Takashi , Yamori Takao , Tsukagoshi Shigeru , Sakurai Yoshio
Published: February 2007

A methotrexate/concanavalin‐A conjugate (MTX/Con‐A) was prepared by covalent cross‐linking. The extent of substitution was approximately 5 moles methotrexate (MTX) per mole concanavalin A (Con A). Inhibition of dihydrofolate reductase by the conjugate was only marginal when compared to MTX. However, MTX/Con‐A showed a 7‐ to 116‐times higher cytocidal activity than MTX against cultured KB cells and various established cell lines from mouse colon adenocar‐cinomas 26 and 38, and Lewis lung carcinoma. The effect of MTX/Con‐A was diminished when the conjugate was preincubated with α‐methyl‐D‐mannoside, a specific binding sugar to Con‐A. MTX/Con‐A efficiently incorporated into cells and retained in them for longer periods of time than was MTX. The strong cytocidal action of the conjugate could be explained by the higher incorporation rate and the longer retention time of the conjugate in the cells.
An analysis of trends in mortality from malignant melanoma of the skin in Australia
Authors: Holman C. D. J. , James I. R. , Gattey P. H. , Armstrong B. K.
Published: February 2007

Australian mortality rates from cutaneous malignant melanoma in successive periods from 1931 to 1977 have been examined with respect to geographic variation and trend with time and birth cohort. The age‐standardized rates rose from 0.8/100,000 in males and 0.6/100,000 in females in 1931–34 to 4.2/100,000 and 2.5/100,000 in 1975–77. Mortality rates were highest in Queensland in the north of Australia and diminished on a gradient from the north to south of the country. An analysis designed to separate effects due to calendar year, birth cohort and age showed that virtually all the secular trend in rates could be explained by increases in successive birth cohorts, beginning as early as 1865 and stabilizing with the cohorts born around 1925 in women and 1935 in men. It is suggested that the cohort‐based increase in mortality resulted from life‐style changes occurring with successive generations. Its stabilization in recent birth cohorts, if persistent, suggests that the secular trend towards increasing total mortality from melanoma will also stabilize over the next 40 years.
In vitro sensitivity of human melanoma xenografts to cytotoxic drugs. Correlation with in vivo chemosensitivity
Authors: Tveit Kjell Magne , Fodstad Øystein , Olsnes Sjur , Pihl Alexander
Published: February 2007

Single‐cell suspensions prepared from five human melanomas, grown serially as xenografts in athymic nude mice, were exposed in vitro to increasing concentrations of DTIC (Dacarbazine), CCNU (Lomustine), procarbazine, vinblastine, and the cancerostatic lectins abrin and ricin. The in vitro chemosensitivity of the cells, as measured by the drug concentrations required to inhibit colony formation in soft agar by 50%, was correlated with the growth delay of the xenografts in vivo, previously observed after treatment of the animals with maximum tolerable doses of the same drugs. It was found that for each drug the in vitro sensitivity of the different xenografts was strongly correlated with their response in vivo. The results provide evidence that the soft agar test, as carried out here, adequately reflects the relative sensitivity of the xenografts in vivo. The data indicate that human xenografts may be used to develop quantitative in vitro chemosensitivity tests.
Two neoplastic cell lines with unique features derived from Hodgkin's disease
Authors: Schaadt Michael , Diehlm Volker , Stein Harald , Fonatsch Christa , Kirchner H. Hartmut
Published: February 2007

Two in vitro cell lines (L428, L439) were established from pleural effusions of two patients with Hodgkin's disease. The histological diagnosis was ascertained in both cases by two independent pathologists. The neoplastic nature of the culture cells is indicated by the demonstration of several structural and numerical chromosome abnormalities associated with a monoclonal pattern of marker chromosomes. Heterotransplantation in nude mice was achieved by intracranial inoculation and by subcutaneous transplantation of cultured cells embedded in a plasma clot. EBV‐specific antigens (EBNA, VCA) were not detectable in either cell line. la‐like antigens, receptors for T cells, acid phosphatase and acid esterase were shown to be present in the cultured cells. The L428 and L439 cell line lacked surface‐ or cytoplasmic 1g, HTLA, receptors for C3b, C3d, 1gG‐Fc, mouse E or sheep E, and were devoid of lysozyme, peroxidase and chloracetate esterase. These features do not correspond to those of B cells, T cells, myeloid cells, monocytes or macrophages; the morphology and the marker pattern of the culture cells, however, is identical with that of freshly obtained Hodgkin (H)‐ and Sternberg‐reed (SR)‐ cells, except for the lack of C1g in the in vitro cells, which is explained by the culture conditions. These findings suggest that the L428 and L439 cell lines are indeed derived from H‐ and Sr‐cells and offer the possibility of gaining new information upon the nature of Hodgkin's disease.
Macrophage content and immunogenicity of C57BL/6J and BALB/cByJ methylcholanthrene‐induced sarcomas
Authors: Evans Robert , Lawler Ellen M.
Published: February 2007

A total of 33 methylcholanthrene‐induced sarcomas, growing in C57BL/6J or BALB/cByJ mice, were tested for their immunogenicity and for their relative mac‐rophage content. No relationship was demonstrated between these two characteristics. These results suggest that the direct correlation between macrophage content and immunogenicity demonstrated in rat tumor model systems may not hold for other systems.
Lung cancer and passive smoking
Authors: Trichopoulos Dimitrios , Kalandidi Anna , Sparros Loukas , Macmahon Brian
Published: February 2007

Fifty‐one women with lung cancer and 163 other hospital patients were interviewed regarding the smoking habits of themselves and their husbands. Forty of the lung cancer cases and 149 of the other patients were non‐smokers. Among the non‐smoking women there was a statistically significant difference between the cancer cases and the other patients with respect to their husbands' smoking habits. Estimates of the relative risk of lung cancer associated with having a husband who smokes were 2.4 for a smoker of less than one pack and 3.4 for women whose husbands smoked more than one pack of cigarettes per day. The limitations of the data are examined; it is evident that further investigation of this issue is warranted.
Chromosome pattern, growth in agar and tumorigenicity in nude mice of four human bladder carcinoma cell lines
Authors: Hastings R. J. , Franks L. M.
Published: February 2007

Four human bladder carcinoma cell lines have been characterized by G‐banding, and the chromosomal patterns correlated to growth in agar and tumorigenicity in nude mice. Each cell line was shown to be chromosomally unique and although numerical and structural anomalies were present, none were common to all four cell lines. However, one or more copies of a structurally altered chromosome 8 were present in all four cell lines and may be associated with tumorigenicity in nude mice. A combination of three marker chromosomes was found in the more anaplastic cell lines, but not in the two well‐differentiated tumour cell lines. Growth in agar may be associated with the presence of the three marker chromosomes but was not correlated with tumorigenicity in nude mice.
Inhibitory effect of cholera toxin on human natural cell‐mediated cytotoxicity and its augmentation by interferon
Authors: Fuse Akira , Sato Takeyuki , Kuwata Tsuguo
Published: February 2007

Cholera toxin inhibits human natural cell‐mediated cytotoxicity in a dose‐ and time‐dependent manner. Pretreatment of lymphocytes with 10 ng/ml of cholera toxin for 2 h almost completely inhibited cytolysis. Interferon augmented human natural cell‐mediated cytolysis, but when lymphocytes were pretreated with cholera toxin before interferon treatment, no enhancement of cytolysis occurred. Cholera toxin could inhibit the enhancement of cytolysis by interferon even when lymphocytes were treated with cholera toxin after 2 h interferon treatment. Cholera toxin subunit B which binds cell surface ganglioside galactosyl‐N‐acetylgalactosaminyl ‐ [N‐ acetylneuraminyl]‐galactosylglucosyl ‐ ceramide (GMI) without activating adenyl cyclase had no effect either on natural cytolysis or on the enhancement of natural cytolysis by interferon, suggesting that mere binding of cholera toxin to the cellular receptor was not enough to inhibit natural cell‐mediated cytolysis. Cyclic adenosine 3′,5′‐monophosphate (cAMP) levels increased in cholera toxin‐treated lymphocytes and the time course of cAMP accumulation was similar to that of cytotoxicity inhibition. Exogenous dibutyryl‐cAMP (db‐cAMP) and theophylline inhibited cytolysis, while exogenous dibutyryl cyclic guanosine 3′,5′‐phosphate (db‐cGMP) enhanced cytolysis slightly, suggesting that the process of inhibition of human natural cell‐mediated cytolysis was at least partly modulated by intracellular cyclic nucleotides.
Characterization of the effects of asparaginase from escherichia coli and a glutaminase‐free asparaginase from vibrio succinogenes on specific cell‐mediated cytotoxicity
Authors: Durden Donald L. , Distasio John A.
Published: February 2007

Asparaginases isolated from Escherichia coli and Erwinia carotovora are effective in the treatment of acute lymphoblastic leukemia in man. During treatment with either of these enzymes, patients frequently experience pronounced toxicity, including liver and pancreatic dysfunction and immunosuppression. The capabilities of these enzymes to hydrolyze L‐glutamine, an important amino acid in mammalian intermediary metabolism, have led investigators to suggest that the glutaminase activity may be responsible for the observed toxicities. Unlike other asparaginases, an enzyme isolated in our laboratory from Vibrio succinogenes, has been shown to be a potent antilymphoma agent and to be highly specific for L‐asparagine. Our previous work has established that the glutaminase‐free asparaginase from V. succinogenes does not suppress the in vivo humoral or cell‐mediated immune responses of mice to sheep red blood cells, whereas the E. coli enzyme abolishes these responses. In this study, we describe the mechanisms by which E. coli asparaginase suppresses specific antibody‐dependent cell‐mediated cytotoxicity. An analysis of the kinetics of appearance of the specific cell‐mediated cytotoxic response of spleen cells after immunization with sheep red blood cells revealed that it paralleled the IgG plaque‐forming cell response. In addition, 18‐h culture supernatants from immune spleen cells conferred upon non‐immune cells the ability to specifically lyse sheep red blood cells. Pretreatment of immune supernatants with protein A‐Sepharose 4B absorbed out the soluble arming factor. An analysis of E. coli asparaginase‐induced suppression of specific antibody‐dependent cell‐mediated cytotoxicity revealed that a decreased synthesis of specific IgG was responsible for the observed reduction in cytotoxic reactivity. When culture supernatants from spleen cells of immunized animals not treated with asparaginase were added to spleen cells from E. coli asparaginase‐treated mice, however, there was an increased cytotoxic response. This suggested that there was an increase in the number or ratio of effector cells for the specific antibody‐dependent cell‐mediated cytotoxic response in spleens of enzyme‐treated animals. Through the characterization of the immunosuppressive effects of two asparaginases—one having the catalytic capability to hydrolyze L‐glutamine and the other lacking this catalytic activity—we now have evidence that the immunosuppressive effects of E. coli asparaginase on specific antibody‐dependent cell‐mediated cytotoxicity cannot be the result of asparagine depletion alone. Our data demonstrate that the glutaminase‐free asparaginase from V. succinogenes is not immunosuppressive and suggest that an asparaginase with high substrate specificity for L‐asparagine may be a safer treatment for leukemia patients.