bluuebunny/crossref_metadata_2025 huggingface dataset:
Radioimmunoassay for infectious primate retrovirus reverse transcriptase: Characterization, comparison with conventional immunologic assays and applicability to cellular extracts
Authors: Robert‐Guroff Marjorie , Kalyanaraman V. S. , Sarngadharan M. G.
Published: February 2007

SSV reverse transcriptase (RT) was purified to homogeneity and used in a radioimmunoassay. Following iodination, the homogeneity of the protein and its identity with RT were confirmed by several criteria: (I) its molecular weight on an SDS‐polyacrylamide gel; (2) its precipitation by anti‐SSV RT but not by antisera to other SSV proteins; (3) its cross‐reactivity in RIA with antisera to other retroviral polymerases; (4) its competition in RIA by active homogeneous SSV RT but not by other purified SSV proteins; and (5) its competition in RIA by only those fractions from a poly(U)‐Sepharose column possessing SSV RT activity. Competition of the labelled probe with disrupted retroviruses of the infectious primate group showed that, while a homologous RIA detected only type‐specific enzyme determinants, it did not distinguish the various woolly‐gibbon retroviral DNA polymerases. A more broadly reactive heterologous assay utilizing an antiserum to R‐MuLV RT detected groupbut not interspecies‐specific enzyme determinants. A comparison of immunologic assays for RT showed that: (I)highly purified RT is not essential for reliable results in enzyme neutralization or enzyme binding assays; (2) the greater sensitivity of enzyme binding compared to enzyme neutralization assays is a function of the antibody, not of the antigen. Competition RIAs using extracts of virus‐infected cells showed that infectious primate retrovirus RT could be measured in a crude system and that cellular DNA polymerares α,β and γ did not compete with the labelled probe.
The binding of benzo(a)pyrene to DNA components of differing sequence complexity
Authors: Jack P. , Brookes P.
Published: February 2007

An examination has been made of the binding, both in vitro and in vivo of the benzo(a)pyrene (BP) adduct to DNA components of differing sequence complexity. Annealing was performed at low renaturation temperatures in the presence of high concentrations of formamide to minimize hydrocarbon‐induced depurination. BHK‐DNA was modified in vitro using a tritiated derivative of the ultimate carcinogenic metabolite of BP, 7α,8β‐di‐hydroxy‐9β,10β‐epoxy‐7,8,9, 10‐tetrahydrobenzo(a)pyrene (BPDE). Co‐renaturation of this modified DNA with [14C]‐thymidine‐labelled BHK‐DNA demonstrated that the hydrocarbon adduct did not interfere with strand annealing and showed that the BP adduct was distributed randomly throughout all DNA sequence classes. However, when the DNA of cells in culture was modified by [3H]BP, following metabolic activation, and mixed with [14C]‐thymidine‐labelled DNA, a small but reproducible difference in the renaturation of the two labels was found. This difference in renaturation profiles was not due to base‐compositional effects since a similar result was found when the alternate 14C‐label was present in guanine bases, the principal site of BP modification. The small difference in the renaturation of the two radioactive labels indicated an enrichment of the hydrocarbon on the most rapidly‐renaturing sequence components (the palindromic and highly repetitive sequences) where it amounted to between 19 and 64% increased modification.
Specific glycolipid antigen(s) in SV40‐transformed cell membranes
Authors: Ansel Sandra , Huet Christian
Published: February 2007

A glycolipid extract was prepared from an SV40‐transformed1 hamster cell line (EH‐SV) according to the Folch partition procedure. The glycolipids from the aqueous layer were incorporated in liposomal membranes composed of lecithin/sphingomyelin/cholesterol (1:1:2 by weight). This liposomal preparation was inoculated in Syrian hamsters to raise immune sera. The sera were absorbed with trypsinized «normal» hamster cells (EH‐N) and tested on various cell lines by the indirect immunofluorescence technique. When used for staining living cells, the immune serum produced a distinct cellsurface fluorescence with SV40‐transformed cell lines regardless of the cell origin (e.g., rat or hamster). No reaction was observed with heterologous Py‐transformed cell lines, spontaneously transformed cells, or sera from non‐immunized hamsters. When used for staining acetone‐fixed cells, the antiglycolipid serum reacted specifically with a thermostable antigen in the nuclear envelope and the cytoplasm of SV40‐transformed cells. The sera lack interfering SV40 T reactivity. The results indicate the presence of related SV40‐specific glycolipid antigen(s) in the plasma membrane, the nuclear membrane and probably other endomembranes of SV40‐transformed cells.
Role of elevated temperature in malignant transformation of mammalian cells in vztro
Authors: Marczynska Barbara , Bergholz Carolyn M. , Wolfe Lauren G.
Published: February 2007

Rat embryo cells subjected in vitro to transient incubation at an elevated temperature (39° C) became transformed and induced fibrosarcomas in both homologous and heterologous hosts. Malignant transformation correlated with the occurrence of karyotypic changes which appeared long after incubation at 39° C and subsequent return to 37° C. Control cultures incubated at only 37° C did not show similar chromosomal changes or induce tumors and remained predominantly diploid during the same observation period. In contrast to rat embryo cells, marmoset monkey cell cultures incubated at 39° C did not develop characteristics of transformed cells.
Dissociation of anchorage independence from tumorigenicity in human cell hybrids
Authors: Stanbridge Eric J. , Wilkinson Joyce
Published: February 2007

The association between anchorage independence and tumorigenicity was examined using a series of intraspecific human cell hybrids. The cell lines represented non‐tumorigenic HeLa/fibroblast hybrids and tumorigenic segregants derived from them. These segregants had lost no more than 5% of the original chromosome complement. Both non‐tumorigenic and tumorigenic cell populations formed colonies in methyl cellulose. The relative size of the colonies seemed to be inherited as a stable trait. Serial cloning of nontumorigenic hybrids in methyl cellulose led to an enhanced efficiency of colony formation but no selection for tumoregenic segregants. Thus, the property of anchorage independence is clearly dissociated from tumorigenicity in this human cell system. An ancillary observation was the chromosomal stability of the hybrids over many population doublings. This intraspecific human cell model therefore provides a genotypically and phenotypically stable system for examination of the genetic control of transformation and neoplasia.
Female sex steroid receptor status in primary and metastatic breast carcinoma and its relationship to serum steroid and peptide hormone levels
Authors: Vihko R. , Jänne O. , Kontula K. , Syrjälä P.
Published: February 2007

In order to obtain more information on the interrelationships between cytosol estrogen (ER) and progestin (PR) receptors in breast carcinoma, and their distribution according to age, menopausal status and endocrine parameters of the patients, these receptors were measured in 605 primary and 150 metastatic lesions, and correlated with serum levels of estradiol, progesterone, FSH, LH and prolactin in some of these patients. Measurable estrogen receptor (> 3 fmol/mg cytosol protein) was found in 78.0% and progestin receptor (> 10 fmol/mg cytosol protein) in 60.5% of all the samples studied. The receptors were simultaneously present in 57.2%, estrogen receptor only in 20.8%, progestin receptor only in 3.3%, while both receptors were absent in 18.7% of the whole material. In samples from 253 premenopausal patients, measurable ER was found less frequently (71.1% of cases) and its concentration was lower (39.9 ± 5.1 fmol/mg cytosol protein, mean ± SEM) than in 502 postmenopausal patients (82%; 148.2 ± 11.6 fmol/mg). The frequencies of ER‐positive samples and ER concentration were rather similar in primary and metastatic lesions, whereas PR was more often present (64 versus 47%) and its concentrations significantly higher (151.2 ± 12.5 versus 102.6 ± 21.1 fmol/mg) in primary than in metastatic tumors. When present simultaneously, there was a significant correlation between ER and PR concentrations in both primary and metastatic lesions independent of the menopausal status of the patient. ER concentration correlated significantly with age in both pre‐ and postmenopausal patients, while PR concentration correlated with age only in postmenopausal patients. The group with the highest ER values (above 100 fmol/mg cytosol protein) had a significantly lower serum estradiol concentration that the other patients. Serum estradiol values had a significantly positive correlation with cytosol PR content in the samples with a measurable PR. Serum progesterone, FSH, LH and prolactin did not correlate with tumor ER or PR concentrations. We conclude that concomitant assays of ER and PR from a breast carcinoma specimen provide a correct picture of the endocrine characteristics of the tumor independently of the serum concentrations of estradiol, progesterone, FSH, LH and prolactin.
The significance of 5′‐nucleotide phosphodiesterase isozymes in the diagnosis of liver carcinoma
Authors: Lu H. M. , Chen C. , Sze P. C. , Ming T. H. , Chiang K. L. , Ting C. R. , Pan L. X. , Liu T. Y. , Yiu C.
Published: February 2007

The results of determination of the serum 5′‐NPDase isozymes in 95 cases of primary liver carcinoma and other kinds of liver disase are presented. The 5′‐NPDase‐V was positive in 83.2% of primary liver cancer cases. This test might be a useful supplement to AFP determination, especially in AFP‐negative liver cancer patients. In most patients who had undergone successful liver resection for primary carconoma, the test became negative. A positive 5′‐NPDase‐V test in patients with cancers elsewhere in the body may suggest liver metastasis. In addition, this test may be of some help in the differentiation of primary liver cancer from other kinds of liver disease. The problem of “false‐positive” results of this test is discussed.
Increase in S‐adenosyl‐L‐methionine decarboxylase activity during the transformation of chick embryo fibroblasts by rous sarcoma virus
Authors: Bachrach Uriel , Wiener Haviva
Published: February 2007

The increase in S‐adenosyl‐L‐methionine decarboxylase activity in chick embryo fibroblasts after infection with Rous sarcoma virus has been studied. It has been shown that enzyme levels in transformed cells were two or three times higher than those of the non‐infected controls. The activity of this enzyme was not elevated in chick embryo fibroblasts infected with a temperaturesensitive mutant of Rous sarcoma virus (RSV‐T5) at 42°C, the non‐permissive temperature. When the temperature of these infected cultures was shifted from 42°C to 37°C a two‐ or three‐fold increase in decarboxylase activity was detected after 10 to 12 h. The half‐live of Sadenosyl‐L‐methionine decarboxylase was practically identical in normal and RSV‐transformed fibroblasts.
Adenovirus type‐12 tumor antigen. III. Tumorigenicity and immune response to syngeneic rat cells transformed with virions and isolated transforming fragment of adenovirus 12 DNA
Authors: Raška Karel , Morrongiello Mary Pat , Föring Bernd
Published: February 2007

Tumorigenicity and immunogenicity of two adenovirus 12 (Ad 12)‐transformed cell lines was studied in syngeneic LIS rats. One cell line (RT2), transformed with Ad 12 virions, contains most of the Ad 12 genome and transcribes its E1, E3 and E4 regions. The other line (RFC1) was transformed with isolated EcoRI‐C fragment of Ad 12 DNA. Both cell lines are equally tumorigenic in newborn rats; in weanling and mature animals the virion‐transformed cells are more tumorigenic as determined by the number of cells required to induce tumors in 50% of animals (TPD50) and by the proportion of “takes” after transplantation of tumor fragments. Serological analysis indicates that antibodies reactive with Ad 12 T‐antigen (product of the E1 region of Ad 12) appear later than those reactive to other Ad 12 antigen(s) present in RT2 cells. The “early” RT2 antibodies are non‐reactive with RFC1 cells. Sera of RT2 tumorbearing rats become reactive with RFC1, cells and isolated Ad 12 T‐antigen after a period of 1–4 weeks. The sera of RFC1 tumor‐bearing rats become reactive with transformed cells after a similar period. The RFC1 sera and the “late” RT2 sera are cytotoxic to both RFC1 and RT2 cells. None of the immunoreactive sera of tumor‐bearing rats are active in antibody‐dependent cellular cytotoxicity (ADCC). The spleen cells of rats bearing RFC1, tumors were cytotoxic to both RFC1 and RT2 cells. The spleen cells of animals with RT2 cell tumors, however, did not exert any cytotoxicity against either cell line. When the spleen cells of tumor‐bearing rats were further educated by incubation with transformed cells in vitro, significant cytotoxicity was observed. The spleen cells of rats bearing RFC1 tumors were equally stimulated in vitro with RFC1 and RT2 cells. The cells of rats bearing RT2 tumors were cytolytic after in vitro exposure to RFC1, cells; much less cytotoxicity was observed with cells restimulated in vitro with RT2 cells. The RFC1 and “late” RT2 sera effectively blocked the cytolysis of both RFC1 and RT2 cells. The “early” RT2 antibodies blocked the cellular cytotoxicity against RT2 cells but not RFC1 cells. Interestingly, the blocking effect was observed even with the effector cells primed in vivo and educated in vitro with only RFC1 cells. These results demonstrate that the E1 gene block of Ad 12 codes for a membrane‐associated antigen active in both humoral and cellular immunity.
Induction of immune reaction against rat embryonal carcinoma by activation of viral genome
Authors: Sobis H. , Van Hove L. , Heremans H. , De Ley M. , Billiau A. , Vandeputte M.
Published: February 2007

A cell line (F3/I) derived from a rat embryonal carcinoma, originally induced in fetectomized rats injected with MSV, was found to be essentially non‐immunogenic in a syngeneic host; the cells did not produce C‐type virions. The MSV genome could be rescued by superinfection of F3/I cells with endogenous C‐type mouse virus and the cells were converted in a producer line‐F3/l‐P. The produced virus was shown to be MSV by its ability to induce focus formation in mouse fibroblasts. In contrast to its non‐producer ancestor, the F3/ I‐P line proved to be strongly immunogenic in the syngeneic host. These results indicate the possibility of converting certain non‐immunogenic tumors into immunogenic ones by the activation of dormant viral genome using xenogeneic non‐oncogenic helper virus.
Time course of C‐type retrovirus expression in mice submitted to osteosarcomagenic doses of 224radium
Authors: Erfle V. , Hehlmann R. , Schetters H. , Meier A. , Luz A.
Published: February 2007

Virus particles with the biochemical properties of C‐type retroviruses appeared transiently in bone tissues of (C3H X IOI)F1 hybrid mice early after treatment with 224Radium; such particles were then again detected in the bones of the irradiated animals at the onset of osteosarcoma formation and in the osteosarcomas. Antibodies against a murine retrovirus isolated from a 224Ratinduced osteosarcoma were produced and detected in the serum of the 224Ra‐treated animals within a month after treatment began. The antibody levels plateaued to a maximum after about 2 months and remained elevated until the tumors started to develop. The antibody concentration in the serum of these irradiated animals decreased then progressively to reach levels similar to those observed in untreated controls. It thus appears that the imminence of osteosarcoma development can be predicted by monitoring the anti‐C‐type virus antibody levels in the serum of the irradiated mice. These experiments also strongly suggest that treatment with 224Radium induces expression of endogenous viruses in the animals.
Epstein‐Barr virus‐specific antibody titers in seven Alaskan Natives before and after diagnosis of nasopharyngeal carcinoma
Authors: Lanier Anne P. , Henle Werner , Bender Thomas R. , Henle Gertrude , Talbot Marion L.
Published: February 2007

Results are presented of Epstein‐Barr virus‐specific serologic tests for seven Alaskan Native patients with nasopharyngeal carcinoma (NPC) from whom serum had been collected 2–10 years before diagnosis and up to 6 years after diagnosis. The pre‐illness antibody spectra and titers of the NPC patients did not differ from those of controls. However, broadened antibody spectra and elevated titers were associated with the emergence of NPC disease, which in one case was present as early as 22 months before actual diagnosis. Increasing or continuously high antibody titers were associated with progression of the disease and death, whereas patients who maintained relatively low, stable antibody titers after treatment have remained well.
Glucocorticoid receptors and glucocorticoid sensitivity of human leukemic cells
Authors: Kontula K. , Andersson L. C. , Paavonen T. , Myllylä G. , Teerenhovi L. , Vuopio P.
Published: February 2007

We have established optimal conditions for the measurement of glucocorticoid receptors (GR) in human white cells using a whole‐cell binding assay with [3H]dexamethasone as the ligand, and the subsequent determination of the GR content in normal human lymphocytes and in leukemic cells of patients with various forms of acute and chronic leukemia. A number of leukemic cell lines in continuous culture were also subjected to the GR assay, and the results were correlated with the sensitivity of these cell lines to glucocorticoid steroids in vitro. The GR content of normal human lymphocytes amounted to 4,850 ± 1,340 (mean ± sd) receptors/cell. The mean equilibrium dissociation constant (KD) of the interaction of [3H]dexamethasone with the GR was 1.2 × 10−8M. Steroidal compounds with a known glucocorticoid potency effectively competed for the binding, whereas steroids devoid of glucocorticoid activity (e.g. estradiol‐17β and testosterone) were ineffective. The GR content of the blast cells obtained from eight patients suffering from acute leukemia and four patients with a blast crisis of chronic myelocytic leukemia was found to be highly variable (3,230–29,900 receptors/cell), while the lymphocytes of six patients with chronic lymphatic leukemia contained a rather stable GR content (2,930–5,120 receptors/cell), which was comparable with that of normal lymphocytes. GR was identified in all the 12 malignant continuous white cell lines studied. Large cells contained more GR than the smaller ones. There was no apparent correlation between the GR concentration and the sensitivity of the cells in vitro to glucocorticoids as judged by [3H]thymidine incorporation studies. Distribution of the surface markers in the leukemic cell lines did not relate to the GR concentration. We conclude that the presence of GR is probably a universal feature of the leukemic cells, and, from a clinical standpoint, probably does not alone imply steroid responsiveness.
Determination of incidence and partial characterization of tumor‐associated antigens found in the urine of patients bearing solid tumors
Authors: Rote N. S. , Gupta R. K. , Morton D. L.
Published: February 2007

Urine samples from patients with solid tumors and from donors without malignant disease were concentrated and tested for the presence of tumor‐associated antigens. In the complement‐fixation assay using serum from a source autologous with the source of the urine, 87.4% of cancer patients were positive, while only 6.9% of control donors were positive. When serum from an allogeneic source was used, 94.7% of cancer patients and 35.1% of control donors were positive. Absorption of a cancer patient's serum with autologous tumor cells removed antibody activity to autologous and allogeneic urine samples. Normal lymphocytes, skin, or muscle‐cell suspensions were ineffective as absorbants. The excretion of antigen into urine is dependent upon the presence of tumor. Surgical removal of tumor resulted in cessation of antigen excretion. The urine remained antigen‐negative as long as the patient remained disease‐free. The antigenic activity was heat‐stable and comprised molecules of > 1,000,000 daltons which could be dissociated into smaller molecular weight active fractions by treatment with 6 m urea.
Induction and specificity of syngeneic antiserum against rauscher murine leukemia virus‐induced leukemia, RBL‐5
Authors: Alaba Olusola , Law L. W.
Published: February 2007

Plasma membranes purified from RBL‐5 leukemia cells and solubilized with Na deoxycholate were fractionated on an Ultrogel AcA34 column. Fractions containing most of the tumor‐rejection activity but low amounts of gp70 and p30 were consolidated and used to hyperimmunize semisyngeneic, CBF1, hybrid mice. Using the complement‐dependent cytotoxicity assay, we determined the antibody titers against RBL‐5 and several other leukemic cells. Only RBL‐5 and EL‐4 tumor cells were lysed. Similar results were obtained when the same tumor cells were used to absorb the syngeneic antiserum and the residual cytotoxic titer determined on RBL‐5 target cells. Coupling of the gamma‐globulins from this antiserum to Sepharose 4B facilitated the isolation of an antigen which inhibited the C'‐dependent lysis of RBL‐5 tumor cells and, when used for immunization, protected the recipient mice against a subcutaneous challenge of RBL‐5 leukemia cells. By means of discgel electrophoresis, in the presence of SDS and 2‐mer‐captoethanol, two polypeptide chains with molecular weights of 46,000 and 21,000 were resolved. The binding of the antigen to a lens culinaris lentil lectin column and its subsequent elution with α‐methyl mannoside suggests that the specific tumor antigen is a glycoprotein.
Macrophage accumulation in primary and transplanted tumors growing in C5‐deficient B10.D2/oSn mice
Authors: Evans Robert
Published: February 2007

The hypothesis was tested that the fifth component of complement (C5) was required for the accumulation of macrophages at the site of tumor growth. This was based on the assumption that the cleavage product of C5, C5a, attracts peripheral blood monocytes along a chemotactic gradient. Methylcholanthrene‐induced primary and transplanted tumors were grown in the genetically C5‐deficient B10.D2/oSn strain and the C5‐positive B10.D2/nSn strain and their macrophage content assessed after enzymic disaggregation of the tumors. The overall data indicated that there were no significant differences in the two strains since macrophages accumulated in the tumors irrespective of the presence of C5 or its cleavage product C5a.
Observations on the EB virus envelope and virus‐determined membrane antigen (MA) polypeptides
Authors: North J. R. , Morgan A. J. , Epstein M. A.
Published: February 2007

Experiments have been carried out to identify the polypeptide components of the Epstein‐Barr (EB) virusdetermined membrane antigen (MA) complex. Cells were radioiodinated using lactoperoxidase and the 125Ilabelled surface antigens, released by Triton X100, were analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS‐PAGE) after complexing either with human sera having anti‐MA activity or with a rabbit antiserum to EB virus. Using cells carrying EB virus isolated from four different conditions, including for the first time nasopharyngeal carcinoma (NPC), we have demonstrated four major polypeptides. One, with a molecular weight of 85,000, was remarkably constant irrespective of the cell line from which it came; two, with molecular weights varying from 240,000 to 270,000 and from 320,000 to 340,000, showed minor differences in mobility apparently depending on the species of origin of the cells (human or marmoset), rather than disparity between strains of virus. A fourth component, of 160,000 daltons, was found on only two of the cell lines studied. In addition, it has been shown for the first time that the same polypeptides composing the MA complex are present on the viral envelope itself. The fact that the rabbit antiserum to EB virus recognized only the two highest molecular weight MA components, yet showed virus‐neutralizing activity, indicates the importance of these two polypeptides for use in a vaccine designed to induce virus‐neutralizing antibodies.
The influence of the lipid composition of the feed given to mice on the immunocompetence and tumour resistance of the progeny
Authors: Boeryd Bernt , Hallgren Bo
Published: February 2007

In inbred CBA mice, the immunocompetence of adult progeny from breeding pairs fed three different diets was compared. Substitution of soy oil for animal fat in the feed of the mice during gestation or lactation significantly decreased the PFC response to SRBC in the adult offspring. Addition of 2‐methoxy‐substituted glycerol ethers to the feed of mothers deprived of animal fat during lactation partly restored the PFC response of the male offspring. In the adult mice fed differently pre‐and perinatally the resistance to a transplanted syngeneic sarcoma was similar. The growth of offspring from mice fed the three diets was similar. In mice deprived of animal fat at weaning and for the following 21 days the immune reactivity to SRBC, tested about 3 months after stopping the diet, was not influenced. However, the resistance to a transplanted tumour in similarly fed mice was increased and this resistance was brought approximately to the control level by methoxy‐substituted glycerol ethers.
Identification of luteinizing hormone‐releasing factor and alpha subunit of glycoprotein hormones in ductal carcinoma of the mammary gland
Authors: Seppälä Markku , Wahlström Torsten
Published: February 2007

The immunoperoxidase technique was used to identify luteinizing hormone‐releasing factor (LRF) and the alpha subunit of glycoprotein hormones in ductal carcinoma of the mammary gland. Eleven tumors were studied, using antisera against LRF and alpha subunit, and against the beta subunits of hCG, LH, FSH and TSH. Six of the tumors were both LRF‐ and alpha‐positive and three were negative. One tumour was LRF‐positive/alpha‐negative, and one was LRF‐negative/alpha‐positive. None of the anti‐beta subunit antisera gave positive staining, and normal breast tissue as well as chronic cystic mastopathy (five cases) were negative with all antisera tested. The results with the anti‐alpha and anti‐beta subunit antisera suggest that the material stained is free alpha subunit rather than any of the native glycoprotein hormones which share the common alpha subunit.
A comparative study of eight cell lines derived from human testicular teratocarcinoma
Authors: Andrews P. W. , Bronson D. L. , Benham F. , Strickland S. , Knowles B. B.
Published: February 2007

Eight independent cell lines, derived from human testicular germ‐cell tumors, were examined for the expression of various markers. These included major histocompatibility and embryonic antigens, chorionic gonadotropin, alpha fetoprotein, alkaline phosphatase, plasminogen activator, and infectivity by SV40. No line consisted primarily of choriocarcinoma or yolk sac cells, but several contained cells resembling murine embryonal carcinoma; some of these lines formed tumors with the distinctive features of embryonal carcinoma when injected into immunosuppressed animals. It is proposed that human embryonal carcinoma cells, unlike those of the mouse, correspond to a preblastocyst stage of development.
Re‐evaluation of a transforming strain of epstein‐barr virus from the burkitt lymphoma cell line, Jijoye
Authors: Sairenji Takeshi , Hinuma Yorio
Published: February 2007

The biological properties of Epstein‐Barr virus (EBV) from a Burkitt lymphoma cell line, Jijoye, were examined. The synthesis of virus capsid antigen (VGA) and early antigen (EA) in Jijoye cells was markedly enhanced by shift‐down of the temperature of incubation from 37°C to 33°C. Cultures of Jijoye cells at 33°C released a large amount of transforming EBV (105.2 of 50% transforming doses/ml) into the culture fluid. However, no infectious virus was produced in all cultures at 37°C during the course of this study. The EBV (Jijoye EBV) from Jijoye line was found to possess only transforming activity, but not EA‐inducing activity. Jijoye EBV lacks adsorbing capacity to Jijoye cells, in contrast to P3HR‐l EBV which can adsorb to Jijoye cells. The Jijoye cells were highly susceptible to superinfection with P3HR‐l EBV as demonstrated by the induction of EA, VGA and infectious EBV. The EBV induced by the P3HR‐I EBV superinfection of Jijoye cells has also transforming activity but neither EA‐inducing activity nor adsorbing capacity to Jijoye cells.
Enhancement of Carcinogenesis in Cultured Respiratory Tract Epithelium by 12‐O‐Tetradecanoylphorbol‐13‐Acetate
Authors: Steele Vernon E. , Marchok Ann C. , Nettesheim Paul
Published: February 2007

The enhancement of chemical carcinogenesis was demonstrated in vitro in rat tracheal epithelium exposed in organ culture to N′‐methyl‐N′‐nitro‐N‐nitroso‐guanidine (MNNG) followed by multiple exposures to 12‐O‐tetradecanoyl‐phorbol‐13‐acetate (TPA). The explants were exposed to 0 or 0.0001 μg MNNG per ml for 6 h on days 3 and 6 of culture, and then to 0 or 1.0 μg TPA per ml for l h on days 9, 15, 21, and 27 of culture. Primary cell cultures were then established from epithelial outgrowths of these explants. Morphologically altered cells were seen in the primary cell cultures derived from MNNG + TPA‐exposed explants on an average of 130 days after MNNG exposure. In primary cultures derived from explants exposed to either TPA or MNNG alone, morphologically altered cells were seen at 190 days and 210 days, respectively. No morphologically altered cells were seen in solvent control primary cell cultures. Only cultures containing morphologically altered cells could be repeatedly subcultured. Cell lines established from these cultures were inoculated i.m. into immunosuppressed isogeneic hosts at 150, 200, 250, 310, and 365 days following carcinogen exposure. All five explants exposed to both MNNG and TPA produced at least one cell line which formed carcinomas when inoculated 365 days following exposure, while only two of the five explants produced tumorigenic cell lines in the MNNG‐only group. Cell lines from explants exposed to both MNNG and TPA were tumorigenic an average of 90 days earlier than cell lines from explants exposed to MNNG alone. None of the cell lines obtained from the explants exposed only to TPA were tumorigenic. These studies show that, following carcinogen exposure, TPA can enhance the oncogenesis of cultured respiratory tract epithelium in terms of increasing the frequency of transformation and decreasing the time at which transformation could first be observed.
Detection of natural killer cells in Japanese quails
Authors: Yamada Akio , Hayami Masanori , Yamanouchi Kazuya , Fujiwara Kosaku
Published: February 2007

Spleen cells of normal quails exhibited cytotoxicity on normal embryo cells as well as on virus‐ and chemically transformed cells. Spleen cells responsible for the destruction of target cells were shown to be non‐adherent lymphoid cells which lack surface immunoglobulin and are resistant to the treatment with anti‐thymocyte serum and complement. Spleen cells of quails which were injected with chicken amniotic fluid (ChAmF) showed no cytotoxic activity. Furthermore, the addition of ChAmF‐treated spleen cells abolished the cytotoxic activity of untreated spleen cells. These results indicated the presence of a spleen‐cell population in normal quails compatible with mammalian natural killer (NK) cells and the induction of suppressor cells directed to the NK cells by administration of ChAmF.
Interferon and spontaneous cytotoxicity in man. V. Enhancement of spontaneous cytotoxicity in patients receiving human leukocyte interferon
Authors: Einhorn Stefan , Blomgren Henric , Strander Hans
Published: February 2007

The effect of daily injections of 3 × 106 units of human leukocyte IF on the spontaneous cytotoxicity of peripheral lymphocytes was studied in 43 patients with a variety of diseases. An increase in spontaneous cytotoxicity was observed 12 to 24 h after the first IF injection with the peak of activity occurring, in most cases, at 24 h. In some patients a decrease in spontaneous cytotoxicity, observed 6 h after the injection, preceded this increase. The increment in spontaneous cytotoxicity was not significantly correlated to preinjection level of cytotoxicity, disease, age, sex, previous therapy, leukocyte counts or proportions of lymphocytes and monocytes. A second injection of IF maintained the spontaneous cytotoxicity at an increased level and during prolonged IF therapy, up to 9 months, the cytotoxicity remained elevated. IF treatment in vitro caused an enhancement of spontaneous cytotoxicity in lymphocytes drawn prior to IF injection. This IF‐induced increase in cytotoxicity in vitro was correlated to the increase in cytototoxicity induced by IF in vivo. Six hours after the first IF injection the ability of IF in vitro to enhance spontaneous cytotoxicity was decreased and during prolonged therapy the ability of IF in vitro to enhance spontaneous cytotoxicity remained decreased.
Estrogen receptor level and other factors in early recurrence of breast cancer
Authors: Westerberg H. , Gustafson S. A. , Nordenskjöld B. , Silfverswärd C. , Wallgren A.
Published: February 2007

At the time of treatment in 1977 for primary breast cancer of all stages, the tumors of 270 women were examined for estrogen‐receptor (ER) content by iso‐electric focusing on polyacrylamide gel. The procedures for the initial and follow‐up examinations were standardized. Postoperative adjuvant radiotherapy or chemotherapy was given to pre‐ and postmenopausal women aged up to 70 years with nodal involvement. Adjuvant tamoxifen treatment (20 mg twice daily for 2 years) was given to one‐half of the postmenopausal patients aged 70 years or less, selected at random, and irrespective of any other adjuvant therapy. These forms of treatment appeared not to introduce bias into the results of the study. There was no correlation between the ER content and tumor spread. Symptom‐free survival during the follow‐up period was best for the patients with tumors with a high ER content, poorer for the patients with tumors of intermediate ER content and least favorable for patients with tumors having low ER content. (Ranges for high, intermediate and low levels: 2.18‐9.08, 0.13‐2.17 and 0‐0.12 fmoles/μg DNA). The differences between the survival curves were significant. For the 120 operable patients with nodal involvement there were also significant differences in recurrence‐free survival. The premenopausal patients in this group likewise showed a significant difference in survival with respect to ER content. For the operable patients with nodal involvement and poorly differentiated tumors there was also a difference in the survival rate depending on the ER content. In this study the ER content of breast cancer was shown to be a quantitative rather than a qualitative entity. The study confirms the independent prognostic importance of the ER content and points to its potential value as a stratification criterion in adjuvant therapy trials.
Polycyclic hydrocarbon activation and metabolism in epithelial cell aggregates prepared from human mammary tissue
Authors: Grover P. L. , MacNicoll A. D. , Sims P. , Easty G. C. , Neville A. M.
Published: February 2007

The metabolism of benz(a)anthracene (BA), 7,12‐di‐methylbenz(a)anthracene (DMBA) and benzo(a)pyrene (BP) by human mammary epithelial cell aggregates in culture has been investigated using non‐neoplastic tissues obtained from eight patients undergoing reduction mammoplasty. All three hydrocarbons were metabolized to water‐soluble and organic solvent‐soluble products and the latter included both K‐region and non‐K‐region dihydrodiols. The major dihydrodiols detected as metabolites of the parent hydrocarbons were the 8,9‐dihydrodiols of BA and DMBA and the 9,10‐dihydrodiol of BP. The 1,2‐dihydrodiols of BA and DMBA and the 11,12‐dihydrodiol of BP were not detected. The hydrocarbons also became bound to the proteins and DNA of the epithelial cells but there were wide differences in the extents of binding occurring with the different hydrocarbons and in the extents of metabolism and binding occurring with tissue preparations from different patients. Some of the hydrocarbon‐deoxyribonucleoside adducts formed from DMBA and BP appeared to have arisen through reactions of »bay‐region« diol‐epoxides with DNA, but only very low levels of reaction with DNA, but only very low levels of reaction with DNA were detected in tissue preparations treated with BA.
The effect of the tumour promoter 12‐O‐tetradecanoyl‐phorbol‐13‐acetate (TPA) on uv‐and MNNG‐induced sister chromatid exchanges in mammalian cells
Authors: Gentil A. , Renault G. , Margot A.
Published: February 2007

The induction of sister chromatid exchanges (SCE) by the tumour promoter 12‐O‐tetradecanoyl‐phorbol‐13‐acetate (TPA) has been studied in V79 Chinese hamster cells comparing control untreated cells with either UV‐irradiated or MNNG‐treated cells. In untreated cells TPA induced SCE at a very low concentration. An increase of approximately 50% over the level of spontaneous SCE formation was observed. In UV‐irradiated cells, TPA, when present after the UV‐irradiation, significantly enhanced UV‐induced SCE formation, more SCE being induced than the expected sum of SCE formed by the separate treatments. TPA also enhanced MNNG‐induced SCE but only when it was present for 28 h before, and not after, the MNNG‐treatment. The protease inhibitor antipain fully inhibited TPA‐induced SCE but did not inhibit UV‐induced SCE. When UV‐irradiated cells were treated with both TPA and antipain the enhanced UV‐induced SCE induction was only partly inhibited by antipain, showing that TPA is responsible for two independent effects, both leading to SCE formation. These results are discussed in relation to the hypothesis that tumour promotion by TPA may depend on the enhancement of mitotic recombination leading to the segregation of a mutational event.
General report on the second international workshop on chromosomes in leukemia
Authors: Rowley J. D.
Published: February 2007

The Workshop participants were able to establish that the t(8;21) and t(15;17) are each definitely associated with a particular type of ANLL, namely, AML‐M2 and APL‐M3 and M3 variant, respectively. Both translocations, particularly the t(15;17), have an uneven geographical distribution which cannot presently be explained. The data also show that patients with preleukemia and an abnormal karyotype have a very poor prognosis with a fatal outcome in 60%, compared with only 30% for patients with a normal karyotype.
Lymphocyte transformation in patients with cutaneous T‐cell lymphomas
Authors: Carney Desmond N. , Bunn Paul A. , Schechter Geraldine P. , Gazdar Adi F.
Published: February 2007

The proliferative responses of peripheral blood mononuclear cells from ten patients with cutaneous T‐cell lymphoma were tested after stimulation with a variety of mitogens, including phytohemagglutinin (PHA), concanavalin A (Con A), pokeweed mitogen (PWM), staphylococcal protein A (SPA), and lymphocyte‐conditioned medium (LyCM). Responses were measured by the incorporation of tritiated thymidine and by flow microfluorometry (FMF). In general, responses were depressed, although a broad range of responses were observed ranging from very poor to near normal responses. SPA and PWM were the most potent mitogens in the CTCL patients, unlike controls in whom PHA and SPA were the optimum mitogens and PWM was weakly mitogenic. FMF studies in two patients clearly demonstrated that aneuploid Sezary cells and not admixed normal lymphocytes were proliferating in response to SPA and PWM stimulation. The unusual pattern of mitogen responses found in these CTCL patients suggests Sezary cells may be derived from T‐cell subpopulations with special affinity for SPA and PWM. The ability of FMF to demonstrate that aneuploid cells respond to mitogen stimulation makes it a useful additional technique in the evaluation of lymphocyte blastogenesis in patients with malignant lymphoproliferative disorders.
Two siblings with acute T‐cell lymphocytic leukemia
Authors: Weber W. , Müller Hj. , Moroni C. , von Fliedner V. , Sartorius J. A. , Speck B. , Obrecht J. P. , Jeannet M.
Published: February 2007

The only two children of clinically healthy parents both developed an acute lymphocytic leukemia of the T‐cell type, one with a mediastinal mass, one without. Extensive laboratory studies revealed a combination of the following unusual circumstances. (I) The injection of leukemic bone marrow into BALB/c‐nu/numice led to an explosive simultaneous development of disseminated lymphomatous tumors with murine karyotype. (2) HLA typing and MLC testing of all four family members revealed sharing of HLA‐A,D and DRw determinants between the parents and pointed to the appearance of suppressor cell activity with the outbreak of the acute leukemia of one sibling. (3) Parental lymphocytes gave a low response to mitogen stimulation, suggesting a subclinical cellular immune defect. It is proposed that the siblings inherited from each parent a defective immune response factor, possibly related to HLA‐D/DRw antigens, that predisposed to acute T‐cell leukemia. The neoplastic process might have been triggered by a transferable agent.
Clonal regulation of the induction of macrophageand granulocyte‐inducing proteins for normal and leukemic myeloid cells
Authors: Falk Avshalom , Sachs Leo
Published: February 2007

A cloned line of myeloid leukemic cells can be induced by the alkylating agent nitrosoguanidine for two macrophage‐ and granulocyte‐inducing (MGI) activities. One activity, MGI‐1, induced the formation of macrophage and granulocyte colonies from normal myeloblasts. Another activity, MGI‐2, induced differentiation of MGI+D+ myeloid leukemic cells to macrophages and granulocytes. Experiments on the time course of induction of the two activities have shown that MGI‐1 was induced before MGI‐2. MGI‐1 was first detected in cell extracts and this was followed by detection of both activities in culture supernatants (conditioned medium). After induction with bacterial lipopolysaccharide, another inducer of both MGI activities in this clone, MGI‐1 was also detected before MGI‐2 in cell extracts. The steroid dexamethasone, which is an effective inducer of some differentiation‐associated properties in this clone, did not induce either MGI‐1 or MGI‐2. Studies with different clones of myeloid leukemic cells have shown a clonal variation in the induction of MGI‐1 and MGI‐2. Different clones were induced by nitrosoguanidine either for MGI‐1 and MGI‐2, for MGI‐1 without MGI‐2, or for neither MGI‐1 nor MGI‐2. None of the clones were induced for MGI‐2 without MGI‐1. The results indicate that the induction of MGI‐1 and MGI‐2 is differently regulated in the same clone, and that there is a clonal and thus presumably genetic variation in inducibility for these two activities of MGI.
A fine fibrous silica contaminant of flour in the high oesophageal cancer area of North‐East Iran
Authors: O'Neill C. H. , Hodges G. M. , Riddle P. N. , Jordan P. W. , Newman R. H. , Flood R. J. , Toulson E. C.
Published: February 2007

We report here the discovery and characterization of a fibrous mineral contaminant of the diet in that area of north‐east Iran where oesophageal cancer has a very high incidence. This contaminant has a smoothly tapering shape and is between 50 and 150 μm long. The greatest diameter is between I and 10 μ and this decreases to a sharply pointed tip with a radius of curvature of between 0.25 and 0.60 μm. Electron microscope X‐ray analysis shows that this fibre consists almost entirely of silica. It is free from alkali metals, aluminium and iron, and therefore differs from other known natural or manmade mineral fibres. Examination of the seeds of more than sixty different species of weed known to contaminate the wheat in this area of the Middle East shows that the fibre originates from the seeds of the common Mediterranean grass Phalaris minor. This seed bears fibres of the same dimensions, composition and birefringence, borne upon the inflorescence bracts which envelop the pericarp of the seeds of this and other members of the Phalaris genus. They are broken off from the seed when the wheat is milled but persist in the flour, where up to 3,000 are found in each gram. Similar fibres can be isolated in quantity from the seeds of related species which are grown commercially, and they have a similar size and composition. When cells of the 3T3 mouse fibroblast line are exposed to these fibres in semi‐solid suspension culture, their proliferation is stimulated more than 100‐fold. We present an hypothesis for the involvement of these plant mineral fibres in the aetiology of oesophageal cancer in Iran and in other areas of high incidence.
The humoral antibody response of rats immunized with chemically modified syngeneic brain cells and glioma cells
Authors: Stavrou D. , Hultén Mariette , Anzil A. P. , Bilzer Th.
Published: February 2007

Six‐month‐old Fischer rats (F344) were given the carcinogen methylnitrosourea in their drinking water. Of the induced brain tumors, four were established in culture and propagated as 78FR‐G‐219 (pleomorphic glioma), 78FR‐G‐299 (astrocytoma), 78FR‐G‐284 and 78FR‐G‐344 (mixed glioma) permanent lines. All cell lines produced S‐100 protein and grew as tumors when inoculated s.c. or i.c. in syngeneic hosts. A comparative study of the antigenicity of these lines at different passage levels was carried out using native and chemically modified cells. Syngeneic rats were immunized with cells conjugated with dimethylsulfate and trinitrobenzene sulfonic acid. The immune response was characterized and quantified by an indirect immunofluorescence method and by a complement‐dependent microcy‐totoxicity test. Chemical modification of the tumor cells enhanced antigenicity of the treated cells. The best results were attained with trinitrobenzene sulfonic‐acidtreated cells and constituted a two‐fold increase in the cytotoxicity index. Cytotoxicity values varied in the different cell lines. Antisera raised with trinitrobenzene sulfonic‐acid‐modified cells of all lines cross‐reacted with cells of all lines. Cytotoxicity values were insignificantly reduced by absorbing the antisera with a variety of syngeneic tissues. Antisera raised against native syngeneic brain cells showed virtually no cytotoxicity for glioma cells. Antisera raised against syngeneic brain cells treated with trinitrobenzene sulfonic acid, however, were slightly cytotoxic for normal brain cells and glioma cells as well. The results of the present studies show that antigenicity of glioma cells can be definitely raised with trinitrobenzene sulfonic acid treatment. Furthermore, it would seem that haptenization of glioma‐associated antigens may be a promising approach to the study of glioma—host interactions.