Showing papers in "International Journal of Cancer in 1969"

Studies on cellular immunity and its serum mediated inhibition in Moloney-virus-induced mouse sarcomas.

Abstract: A colony-inhibition technique was used to demonstrate lymph-node cell (LNC) mediated immune reactions against tumor-specific transplantation antigens of primary, Moloney sarcoma virus induced mouse sarcomas. Lymph-node cells from mice in which Moloney sarcomas had regressed spontaneously (regressors), as well as LNC from mice carrying progressively growing tumors (progressors), induced by virus inoculation at an age of 14 days or older, reduced the plating efficiency of Moloney sarcoma target cells. Serum from mice with progressively growing Moloney sarcomas, but not from mice with spontaneous mammary carcinomas or methylcholanthrene-induced sarcomas, abrogated the inhibitory effect of regressor LNC on Moloney sarcoma cells. Additional evidence for the specificity of the serum effect was obtained in experiments showing that the protective effect of progressor sera could be specifically removed by absorption with Moloney sarcoma cells. Regressor sera gave no protection against target cell inhibition by regressor LNC. It is suggested that progressor sera contain antibodies which mediate an efferent form of immunological enhancement. Inhibition Serique de l'Immunite a la Tumeur de Moloney On a utilse une technique d'inhibition des colonies pour mettre en evidence les reactions immunitaires des cellules des ganglions lymphatiques (CGL) contre les antigenes spcifiques de transplnatation de sarcomes primitifs de souris provoques par le virus du sarcome de Moloney. Les CGL des souris chez lesquelles le sarcome de Moloney avait spontanement regresse (“regresseurs”) et les CGL des souris porteuses de tumeurs a croissance progressive (“progresseurs”) induites par inoculation de virus a l'ǎge de 14 jours ou plus ont abaisse la vitalite des colonies de cellulescibles du sarcome de Moloney. Du serum de souris presentatnt des sarcomes de Moloney croissant progressivement — a la difference du serum de souris atteintes d'epitheliomas mammaires spontanes ou de sarcomes provoques par le methylcholanthrene — a supprime l'effet inhibiteur des CGL “regesseurs” sur les cellules du sarcome de Moloney. On a obteny une preuve supplementaire de la specificite de l'effet du serum par des experiences montrant que l'effet du serum par des experiences montrant que l'effet protecteur des serums “progresseurs” pouvait ětre specifiquement supprime par absorption au moyen de cellules du sarcome de Moloney. Les serums “regresseurs” n'ont donne aucune protection contre l'inhibition des cellules-cibles par les CGL “regresseurs”. On peut avancer l'hypothese que les srums “progresseurs” contiennent des anticorps (ou des fragments d'anticorps) mediateurs d'une forme efferente de facilitation immunologique ou eventuellement de modulation antigenique.

A mathematical model for the age distribution of cancer in man

Abstract: Incidence data that have been collected by cancer registries in different parts of the world provide an opportunity for examining the relationship between cancer incidence and age on a wider scale and more accurately than has been possible before. In the present paper the relationship has been examined in a large group of cancers whose incidence increases progressively from young adult life into old age. Five-year age-specific incidence rates between 35 and 74 years of age were recorded for 31 types of cancer (22 in men and 9 in women) in 11 populations, and the median age of each age group was used to characterize the group. The data were used to find the best fitting constants for each type of cancer in each population, in the equation: where I is the incidence at age t, and b and k are constants. With this equation a straight line is obtained, when incidence and age are plotted on a double logarithmic scale. On visual examination, 21% (72 out of 338) of the sets of data appeared to fit the relationship closely. In 54% (181 out of 338) the lines showed downward curvature; that is, the rate of increase with age was less than predicted by the equation; and in 25% (85 out of 338) the lines showed upward curvature. In one-third of the total the differences were too great to be readily attributable to chance. An alternative hypothesis was examined which postulated that the risk of cancer was determined, not by the age of the subject, but by the prevalence of carcinogenic agents and the length of time the subject had been exposed to them. The equation was then written: where w is a constant and t-w represents the ‘effective exposure’ between first exposure and the first appearance of cancer as a pathological entity. The data for cancer of the prostate were found to fit the equation adequately in all 11 populations, when w was 32.5 years. No evidence was found to suggest that the shape of the observed relationship could be attributed to attenuation of a limited pool of susceptibles. Changes in prevalence of carcinogenic agents could account for the variation in the pattern of the observed relationship, and the conditions under which this might occur are discussed. Differences in the estimated value of k (the power of the effective exposure time in the equation), were examined between (i) types of cancer and (ii) populations. The results were not wholly consistent but suggested that the value of k might be a biological constant characteristic of the tissue in which the cancer is produced. Un Modele Mathematique de la Distribution par Ages du Cancer chez l'Homme Les donntes recueillies sur I'incidence dans Ies Jichiers du cancer de diffrentes rdgions du monde permettent d'examiner, plus amplement et plus exactement qu'on nepouvait Ie faire auparavant, Ia relation entre I'incidence du cancer et I'rige des malades. Dans le prksent article, cette relation a ete ittudite pour un grand nombre de cancers dont I'incidence augmente progressivement du dtbut de I'rige adulte jusqu' ´ la vieillesse. On a releve les taux d'incidence par tranches de cinq ans entre les âges; de 35 ´ 74 ans pour 31 types de cancer (22 pour le sexe masculin et 9 pour le sexe feminin) dans 11 populations, et I'on a utilise pour caracteriser chaque groupe d'âges I'âge median du groupe. On a cherche d'apres les donnees a determiner les constantes s'adaptant le mieux a chaque type de cancer dans chaque population, dans I'equation: Dans cette equation, I est l'incidence a I'âge t, et b et k sont des constantes. Avec cette equation, on obtient une droite si I'on trace la courbe de l'incidence en fonction de I'âge avec des coordonnees logarithmiques. D'aprh I'examen des courbes, 21% (72 sur 338) des series de donnees paraissaient correspondre etroitement a cette relation. Dans 54% des stries (181 sur 338) les courbes s'inclinaient vers le bas, autrement dit le taux d'accroissement en fonction de I'âge etait inferieur a celui que prevoyait I'equation; et dam 257; des series (85 sur 338), Ies courbes s'inclinaient vers le haut. Dans un tiers des series 1es differences etaient trop grandes pour qu'on puisse avec vraisemblance les imputer au hasard. On a examine une autre hypothese postulant que le risque de cancer etait determine non par I'âge du sujet mais par la prevalence des agents carcinogenes et par la duree prealable d'exposition du sujet a ceux-ci. L'equation devenait alors: Dans cette equation, w est une constante et t-w represente ‘ I'exposition eflertive ’ entre le debut de I'exposition et I'apparition du cancer en tant qu'entite pathologique.

Quantitative studies on the malignant transformation of mouse prostate cells by carcinogenic hydrocarbons in vitro.

Abstract: A quantitative system for producing malignant transformation of adult C3H mouse ventral prostate cells in culture with carcinogenic hydrocarbons has been developed. Aneuploid cell lines derived from mouse prostate are plated at a low density on a feeder layer and treated with various hydrocarbons for different lengths of time; other dishes are treated with 0.5% dimethyl sulfoxide (the solvent for the hydrocarbons) as controls. After 8 days some dishes are fixed and stained and the colonies are counted for total plating efficiency. Two weeks later other dishes are also fixed and stained. In the DMSO controls, the cells remain as a monolayer. In the carcinogen-treated dishes, piled-up multilayered colonies are seen and their frequency is scored. It has been shown that each of such piled-up colonies gives rise to fibrosarcomas in mice, whereas the cells obtained from the monolayer areas of treated dishes do not cause tumors. Thus, these piled-up colonies represent malignant transformation, under conditions where no spontaneous malignant transformation occurs. There is an excellent quantitative correlation between the carcinogenic activities of eight hydrocarbons and the transformation frequencies they produce in this system. Malignant transformation has been produced by treatment with methylcholanthrene for only one day. The acquisition of heat sensitivity is an early consequence of malignant transformation in this system.

Lactation and reproductive histories of breast cancer patients in greater Athens, 1965-67.

Abstract: As part of an international collaborative study, an attempt was made to identify all cases of breast cancer among residents of the Cities of Athens and Piraeus newly diagnosed in a period of 2 1/2 years. A total of 956 cases was identified, giving an annual average incidence rate of 34.9 per 100,000 female population. Rates by age and marital status are reported. Of these cases 799 (84%) were interviewed, along with 2,470 controls. The controls were residents of Athens and Piraeus under treatment in the same hospitals as the breast cancer cases but for conditions other than breast cancer. In relation to the controls, the breast cancer cases were of low fertility. After adjustment for this difference there was no difference between cases and controls in any of a variety of measures of lactation experience. The cases differed significantly from the controls in having higher socio-economic status, earlier menarche, later first pregnancy, later menopause and greater height and weight. The ranges of these variables were associated with variations in breast cancer risk of the order of two or three fold.

Transplantable mouse tumor line induced by injection of SV40‐transformed mouse kidney cells

Abstract: A transplantable tumor of inbred mice was obtained by inoculating BALB/c mice subcutaneously with SV40-transformed mouse kidney (mKS-A) cells. Tumors were produced by mKS-A cells in the 71st cell culture passage, but not by cells in the 26th passage. The tumor line has been serially passed in BALB/c mice 14 times. In vitro cell culture lines were derived from tumors after 1, 2, 8, 10 and 12 passages in mice. The tumors, as well as the In vitro tumor cell lines, contained SV40 T-antigen, and sera from the tumor-bearing mice contained antibodies to the SV40 T-antigen. SV40 was rescued from the In vitro tumor cell lines after fusion with green monkey kidney (CV-1) cells in the presence of UV-irradiated Sendai virus. The In vitro tumor cell lines derived from mouse passages 8, 10 and 12 were used as SV40 virus; 2) SV40-transformed cell lines; 3) primary mouse (BALB/c or Yale Swiss) kidney cells, or 4) primary mouse (BALB/c or Yale Swiss) embryo cells. These results showed that the tumor line and the In vitro tumor cell lines have the transplantation antigen.

Metabolism and binding to cellular macromolecules of a series of hydrocarbons by mouse embryo cells in culture.

Abstract: A series of eight tritium-labelled polycyclic hydrocarbons were incubated in the presence of monolayer cultures of primary mouse embryo cells, and the time-course of their metabolism to water-soluble derivatives was studied. All the hydrocarbons, both carcinogenic and non-carcinogenic, were metabolized at approximately the same rate when a low concentration of the order of 0.05 μM was used. The binding of the various hydrocarbons to cellular DNA, RNA and protein was determined and a “binding index” calculated, i.e., the extent of reaction of the hydrocarbon with the particular macromolecule resulting from the metabolism of 1 mμmole of hydrocarbon per ml of medium to which the cells were exposed. While no very significant difference between the various hydrocarbons was found for the protein binding index, the DNA and RNA binding indices divided the compounds into two groups. One group with a high binding index consisted of potent carcinogens while the other group had much lower (one tenth) values for the binding index. With the exception of dibenz[a, h]anthracene, this latter group consisted of non-carcinogens.

Cellular immunity and its serum-mediated inhibition in shope-virus-induced rabbit papillomas

Abstract: A colony-inhibition technique was used to demonstrate lymph-node-cell (LNC) mediated immune reactions against tumor-specific transplantation antigens of Shope-virus-induced papilloma and carcinoma cells. Lymph-node cells from rabbits in which Shope papillomas had regressed spontaneously, as well as LNC from rabbits with persistent Shope papillomas, reduced the plating efficiency of Shope-virus-induced papilloma and carcinoma cells but did not affect normal skin fibroblasts from the same rabbits as the tumor cells. Serum from rabbits with persistent Shope papillomas or Shope carcinomas abrogated the inhibitory effect of regressor LNC on Shope tumor cells, while serum from regressor rabbits had no such effect. It is postulated that sera from “persistor” and carcinoma rabbits contain antibodies which mediate an efferent form of immunological enhancement. Inhibition Serique de l'Immunite. A la Tumeur de Shope On a utilise une technique d'inhibition cles colonies pour mettre en evidence les reactions inimunitaires des cellules dr ganglions lytiiphatiques (CGL) contre les antigenes specifiques de transplantation de cellules d'epithkliotiius et de papillotnes provoques par Ie virus de Shope. Les CGL dc lapins chez lesquels les pupillomes de Shope avaient regresse spotitanement, ainsi que les CGL de lapins presentant des papillomes de Shope persistants, ont abaisse la vitalite des cellules des epithelionias et des papillomes induits par le virus de Shope, mais n'ont pas affecte les fibroplastes cutanes normaux des lapins qui presentaient ces cellules tumorales. Du serum de lapins chez lesquels les papillomes ou les epitheliomas de Shope persistaient a annule l'effet d'inhibition des CGL “regresseurs” sur les cellules des tumeurs de Shope, alors que le serum de lapins “regresseurs” n'avait pas cet effet. On estime que le serum des lapins “persisteurs” et porteurs d'epitheliomas contient des anticorps mediateurs d'une forme efferente de facilitation immunologique.

Studies on serum-mediated inhibition of cellular immunity to spontaneous mouse mammary tumors.

Abstract: Sera from mice which have either spontaneously developed a mammary tumor, or which have been immunized against such a tumor, have been shown capable of abrogating the ability of lymph-node cells from these mice to specifically inhibit colony formation by mammary tumor cells in vitro. Sera from mice sensitized to another, independently arising, syngeneic mammary tumor or to a syngeneic Moloney sarcoma virus-induced tumor are less active in this respect, indicating that this reaction is specific. It is suggested that these observations are related to the in vivo phenomenon called immunological enhancement. Inhibition Serique de l'Immunite des Tumeurs Mammaires On a observe que des serums de souris chez lesquelles une tumeur niamniaire est apparue spontanement ou qui ont ete immunisees contre ce type de tumeur ont pu empecher Ies cellules de ganglions lymphatiques de ces souris d'inhiber specifquenient la formation de colonies par des cellules de tumeurs mammuires in vitro. Les serums de souris sensibilisees a une autre tumeur mammaire syngeneique apparaissant independamment, ou a une tumeur syngeneique provoquee par le virus du sarcome de Moloney, sont moins actifs a cet egard, ce qui indique qu'il s'agit d'une reaction specifique. II est possible que ces observations soient en rdation avcc le phenomene in vivo appele „facilitation immunologique”.

Relationship between previous reproductive history and chemically induced mammary cancer in rats

Abstract: The relationship between a previous history of multiple pregnancies and/or lactations and 7,12-dimethylbenz(a)anthracene-induced mammary carcinogenesis was studied in groups of Sprague-Dawley rats. All rats received the carcinogen at approximately 6 1/2 months of age. Virgin control rats which did not receive the carcinogen did not develop mammary cancer within 13 months although carcinogen-fed virgin rats exhibited a mammary cancer incidence of 38.5%. A single pregnancy prior to feeding the carcinogen resulted in a decrease in the incidence and an increase in the mean time of appearance of the first palpable tumor. A single pregnancy followed by a subsequent nursing period reduced the percentage incidence and increased the mean time of appearance of the first palpable tumor when compared to the carcinogen-fed virgin rats. A further increase in the number of pregnancies and/or lactations did not greatly influence the incidence of mammary cancer nor the number of cancers per rat. The mean time of tumor appearance was, however, increased. These data indicated that the relationship between party and carcinogen-induced mammary cancer in the rat is similar to that existing in the human female but opposite to that found for the mouse.

Identification of the filtrable leukocyte-transforming factor of QIMR-WIL cells as herpes-like virus.

Abstract: The QIMR-WIL human leukocyte cell line contains a filtrable factor which transforms foetal human leukocytes and allows establishment of cell lines. The factor resembled members of the Herpesvirus group in its size as determined by filtration and in its sensitivity to high temperature and ether. The transforming activity of the factor was completely neutralized by human sera containing antibody to the herpes-like virus of the QIMR-WIL line but not by control human sera without this antibody. It was concluded that the QIMR-WIL transforming factor was the herpes-like virus. Similar transformation was effected by filtrates of the Burkitt lymphoma line QIMR-GOR which also carries herpes-like virus. The delayed onset of leukocyte proliferation occurring with virus from QIMR-GOR cells (49 days) compared with virus from QIMR-WIL cells (23 days) suggests that strain differences exist.

The epidemiology of breast cancer: review and prospects.

Abstract: In the epidemiology of a disease, which is multifactorial to such an extent as we believe breast cancer to be, attention should be paid even to “weak” factors and an attempt should be made to assign a place to them in an aetiologic system which is biologically sound. Medical knowledge will help us in doing this, and the exercises in “metabolic epidemiology” will be as useful as statistical manipulations and computer programming. Two developments seem to be taking place, one of them being the teamwork between analytic epidemiology and biochemistry; the other is that we are begining to realize that most so-called chronic diseases of relatively high prevalence are like icebergs in the sea with a vast amount of subclinical abnormalities in the “healthy” population (and therefore in our control groups). Epidemilogie du Cancer du sein: Analyse et Perspectives Dans l'epidemiologie d'une maladie a multiples facteurs, comme l'est a notre avis le cancer du sein, il faudrait s'interesser aussi aux facteurs secondaires et essayer de les integrer a un systeme etiologique qui soit biologiquement valable. Nos connaissances medicales nous aideront a le faire et les exercices d' “epidemiologie metabolique” seront aussi utiles que les manipulations statistiques et l'utilisation d'ordinateurs. Une evolution semble en cours, dont l'un des elements est l'esprit d'equipe qui caracterise les travaux de biochimie et d'epidemiologie analytique; en outre, nous commencons a prendre conscience du fait que la plupart des maladies que l'on dit chroniques et dont l'incidence est relativement elevee ressemblent a des icebergs flottant sur la mer et s'accompagnent d'une grande quantite d'anomalies sub-cliniques dans la population “saine” (et par consequent dans nos groupes-temoins).

The formation of variants with a reversion of properties of transformed cells. III. Reversion of the structure of the cell surface membrane

Abstract: The cell surface structure of variants from polyoma-transformed cells grown in animals and then cultured in vitro, which show a reversion of in vitro properties of transformation without losing the ability to synthesize the polyoma-specific nuclear tumor (T) antigen, has been studied by using the carbohydrate-binding protein concanavalin A (Con. A) which interacts with sites on the cell surface membrane. Transformed cells are agglutinated by this protein. Normal cells, whose sites for Con. A are mainly in a cryptic form, are agglutinated only after they have been treated with trypsin. It has been shown that some variants showed a partial, and others a complete, loss of agglutinability by Con. A, and that the agglutinability of these variants was restored by treatment with trypsin. Compared to the parental transformed cells, all the variants showed the same decrease in cloning efficiency in fluid medium and soft agar, and in saturation density. The results indicate that the surface structure of transformed cells can completely revert to the structure found in normal cells, as measured by reversion to the cryptic form of sites for Con. A; that the synthesis of T antigen, presumably due to integrated virus DNA, is by itself not sufficient to prevent this reversion; that variants with this reversion in cells grown in vitro can have a high degree of tumorigenicity in animals; and that a threshold for the number of exposed surface sites for Con. A has to be exceeded before the cells express the in vitro properties of transformation. Reversion de la Surface des Cellules Transformees La structure de la surface cellulaire de mutants de cellules de polyome transformees prelevees sur des animaux puis cultivees in vitro — ou l'on observe une reversion des proprietes de transformation in vitro ainsi que la synthese de l'antigene (T) des tumeurs nucleaires specifiques du polyome — a ete etudiee au moyen de la concanavaline A (Con. A) — proteine de liaison glucidique — qui agit sur les sites de la membrane de la surface cellulaire. Les cellules transformees s'agglutinent en presence de cette proteine. Les cellules normales, dont les sites reagissant a la Con. A sont generalement de forme cryptique, ne s'agglutinent qu'apres un traitement a la trypsine. Il a ete demontre que certains mutants presentent une perte partielle et parfois totale d'agglutinabilite par la Con. A, et que l'agglutinabilite de ces mutants est retablie par traitement a la trypsine. Par rapport aux cellules meres transformees, tous les mutants presentent la měme diminution de l'efficacite de clonage dans un milieu liquide et dans la gelose molle, ainsi que de la densite de saturation. Les resultats montrent 1) que la structure de la surface des cellules transformees peut redevenir exactement la měme que celle des cellules normales, comme on l'a mesure par retour a la forme cryptique des sites reagissant a la Con. A; 2) que la syntheese de l'antigene T, sans doute due a l'ADN a virus integres, ne suffit pas en elle-měme a empěcher cette reversion; 3) que les mutants presentant cette reversion dans des cellules cultivees in vitro peuvent ětre fortement tumorigenes chez l'animal; et 4) qu'il faut depasser un seuil — quant au nombre de sites de surface exposes a la Con. A — avant que les cellules acquierent les proprietes de transformation in vitro.

EBV-associated serological patterns in a Burkitt lymphoma patient during regression and recurrence.

Abstract: Three serological reactions have been followed in a Burkitt lymphoma patient during the clinical course of the disease, including long‐term regression, followed by recurrence and progressive growth, in spite of chemotherapy. Antibodies directed against surface antigens characteristic for EBV‐carrying lymphoblastoid cell lines of Burkitt lymphoma origin were assessed by blocking of direct membrane immunofluorescence. The level of surface reactive antibodies was high while the patient was in long‐term regression. It decreased to an insignificant level approximately half a year before tumor recurrence was clinically diagnosed. Immediately after the recurrence, it remained at a low level but increased subsequently and remained at a high level until the death of the patient. The anti‐EBV antibody level itself showed only minor fluctuations. Precipitating antibodies, directed against soluble antigen extracted from the EBV‐carrying Jijoye cell line, were absent during long‐term regression. They appeared soon after recurrence and remained at a high level until the patient died. Repeated biopsy specimens were taken after the recurrence. In the course of progressive growth, a coating of the IgG type was seen to accumulate on the surface of the tumor cells. Possible implications of these findings are discussed.

Serum α1 foeto-protein (fetuin) in children with malignant ovarian or testicular teratomas. Preliminary results

Abstract: In a study of sera of 93 children bearing various malignant tumours, the authors were able to detect by an immunological test an enthryonal protein, α1 foeto-protein or fetuin, in one testicular and four ovarian embryonal carcinomas. Children bearing tumours other than malignant teratomas, and control children, gave negative results (with one exception in a normal 3-day-old infant). Such an immunological test, therefore, appears to have some diagnostic and prog nostic value which deserves further investigation. Une Foeto-Proteine (Fetuine) Serique chez des Enfants Atteints de Teratome Ovarien ou Testiculaire Malin – Resultats Preliminaires Au cours d'une etude sur le serum de 93 enfants atteints de diverses tumeurs malignes, les auteurs ont pu mettre en evidence par une epreuve immunologique une proteine embryonnaire (foeto-proteine α1 ou fetuine) dans quatre epitheliomas embryonnaires de l'ovaire et un du testicule. Les resultats etaient negatifs chez les enfants atteints de tumeurs autres que des teratomes malins et chez les temoins (a l'exception d'un enfant normal ǎge de 3 jours). Cette epreuve immunologique parait donc avoir une certaine valeur diagnostique et pronostique qui merite une etude plus approfondie.

Localisation cellulaire d'un antigène embryonnaire de tumeurs coliques humaines

Abstract: Une etude microscopique en immunofluorescence a ete entreprise pour localiser l'antigene carcino-embryonnaire (ACE), isole auparavant de tumeurs coliques humaines. Des coupes a congelation de tissu peritumoral, non tumoral, de muqueuse colique normale et d'intestins fœtaux ont ete examinees en utilisant la methode indirecte de Coons. Des images caracteristiques de fluorescence ont ete observees lorsque l'immun-serum anti-antigene carcino-embryonnaire (ACE) a ete employe. Dans les tumeurs et dans l'intestin fœtal, les membranes libres des cellules bordant la lumiere des glandes sont devenues fortement fluorescentes revelant ainsi la presence de l'ACE. Quand l'antiserum est absorbe par l'ACE, cette reaction disparaǐt. Des contrǒles avec des antiserums differents ne donnent pas les měmes resultats. Ceci est en faveur de la specificite des images obtenues. Cellular localization of an embryonic antigen in human colonic tumors A study by immunofluorescence microscopy was undertaken to localize a carcinoembryonic antigen (CEA), which was previously isolated from human colonic tumors and which has been found to be identical to that described first by Gold and Freedman in 1965. Frozen sections of tumors, peritumoral, non tumoral, and normal colonic mucosa, as well as fetal intestines, were examined by the indirect method of Coons. Distinct immunofluorescent patterns were found if specific anti-CEA antiserum was employed. The free membranes of the cells bordering the lumina of the glands became strongly fluorescent in sections of tumors as well as in sections of fetal intestine, thus indicating the presence of CEA. When the antiserum was absorbed with CEA, this reaction was abolished. The specificity of the reaction for CEA was further substantiated by the observation that control antisera produced a distinctly different pattern of immunofluorescent staining.

In vitro demonstration of tumor‐specific antigens in spontaneous mammary tumors of mice

Abstract: The colony-inhibition test has been used to demonstrate cellular immunity against spontaneous murine mammary tumors, both in autochthonous, mammary tumor virus (MTV)-carrying BALB/cfC3H and in immunized, MTV-free, BALB/c mice. Immunity could be demonstrated in nearly all cases against a mammary tumor with lymph-node cells from mice which had experienced that tumor. One of eight tumors tested with lymph-node cells from BALB/cfC3H mice which had spontaneously developed mammary tumors showed evidence of immunological cross-reactivity, while seven of thirteen tumors did so when tested with immune cells from MTV-free BALB/c mice. The data, therefore, indicate the presence of at least two types of tumor-specific antigen (s) in mouse mammary tumors — a common one and an antigen specific for each neoplasm.

Studies on plasma membranes. IX. A survey of enzyme activities displayed by plasma membranes isolated from normal and preneoplastic livers and primary and transplanted hepatomas of the rat

Abstract: Plasma membranes were isolated from normal rat livers, livers of rats fed for various periods of time with 4-dimethylaminoazobenzene (4-DAB), primary hepatomas, and transplants of hepatoma 484. The tissues were homogenized in three media, i.e. 1 mM HCO3 of pH 7.5, 1 mM NaHCO3 containing 2 mM CaCl2, and 2.8 mM citric acid. The isolated plasma membranes were assayed for enzymes previously demonstrated in plasma membranes isolated from rat-liver homogenates prepared in 1 mM NaHCO3. Enzyme levels in liver membranes obtained from dilute citric acid homogenates differed markedly from those obtained following tissue homogenization in the bicarbonate media. The hepatoma-484 membranes obtained from citric acid homogenates resembled enzymatically those isolated from bicarbonate-CaCl2 medium. Of the three homogenization media employed, 1 mM NaHCO3-2 mM CaCl2 (CaCl2 being required for the isolation of hepatoma-484 membranes, but being optional for liver) yielded membranes most suitable for a comparative investigation. None of the enzymes present in liver membranes was absent from the hepatoma-484 membranes. Hexokinase was present in the latter but absent from liver and primary-hepatoma membranes. No difference in specific enzyme activities between liver and hepatoma-484 membranes was observed for acid nitrophenylphosphatase, acetylphosphatase, K+-acetylphosphatase, glucose-6-phosphatase and, as formerly shown, leucyl-β-naphthylamidase. Alkaline and acid glycerolphosphatases, alkaline nitrophenylphosphatase and alkaline K+-nitrophenylphosphatase, alkaline and acid phosphodiesterases, and (Na±K+) ATPase were from slightly to markedly increased in the hepatoma as compared with liver membranes, The ATPase, 5′-mononucleotidase, “neutral” nitrophenylphosphatase and “neutral” K+-nitrophenylphosphatase, inosine diphosphatase and non-specific esterase activities were lower in hepatoma than in liver membranes. Alkaline glycerolphosphatase and acid phosphodiesterase increased and ATPase and non-specific esterase decreased in activity with the development of the neoplastic process to reach, respectively, the highest and the lowest levels in the membranes of the transplanted hepatoma. Enzymes des Membranes de Plasma d'hepatomes de rat Des membranes de plasma ont ete isolees sur des foies normaux de rats, des foies de rats auxquels a ete administre pendant certaines periodes du dimethyl-4-aminoazobenzene (4-DAB), des hepatomes primitifs et des transplants d'hepatomes-484. Les tissus ont ete homogeneises dans trois milieux, soit I mM de HC03 de pH 7.5, I mM de NaHCO3 contenant 2 mM de CaCl2, et 2.8 mM d'acide citrique. Les membranes de plasma isolees ont ete analysees pour la recherche d'enzymes deja observes dans des membranes de plasma isolees sur des homogenats de foies de rat prepares dans I mM de NaHC03 Les niveaux d'enzymes dans les membranes hepatiques obtenus a partir d'homogenats dans I'acide citrique dilue etaient tres diferents de ceux que I'on avait obtenus apres homogenetisation tissulaire en milieu bicarbonate. Les membranes d'hepatome-484 obtenues a partir d'homogenats dans I'acide citrique ressemblaient, du point de vue enzymatique, a celles qui avaient ete isolees dans un milieu bicarbonate du CaCl2. Des trois milieux d'homogeneisation utilises, I mM de NaHCO3 et 2 mM de CaCl2, (CaCl2, etant necessaire pour isoler les membranes d'hepatome-484, mais facultatif pour le foie) ont donne les membranes les mieux utilisables pour une recherche comparee. Aucun des enzymes presents dans les membranes hepatiques n'etait absent des membranes d'hepatome-484. L'hexokinase etait presente dans ce dernier mais n'apparaissait pas dans les membranes du foie et des hepatomes primitifs. On n'a observe aucune difference dans l'activite enzymatique specifique des membranes de foie ou d'hepatome-484 en ce qui concerne la nitrophenylphosphatase acide, l'acetylphosphatase, l'acetylphosphatase-K+, la glucose-6-phosphatase et, comme on l'a montre precedemment, la leucyl-β-naphtylamidase. Les glycerolphosphatases acide et alcaline, la nitrophenylphosphatase alcaline et la nitrophenylphosphatase-K+ alcaline, les phosphodiesterases alcaline et acide, et la (Na+-K+) ATPase apparaissaient en quantite fortement ou legerement accrue dans l'hepatome, par comparaison avec les membranes hepatiques. L'activite de l'ATPase, de la mononucleotidase-5′, de la nitrophenylphosphatase “neutre” et de la nitrophenylphosphatase-K+ “neutre”, de la diphosphatase inosine et de l'esterase non specifique etaient plus faibles dans les membranes de l'hepatome que dans celles du foie. L'activite de la glycerolphosphatase alcaline et de la phosphodiesterase acide a augmente alors que celle de l'esterase non specifique a diminue avec l'evolution du processus neoplasique, pour atteindre respectivement les niveaux maximaux et minimaux dans les membranes de l'hepatome transplante.

Synthesis and fate of immunological surface receptors on cultured burkitt lymphoma cells

Abstract: The distribution and rate of appearance of newly synthesized receptors on the surface of Burkitt lymphoma cells were determined by membrane immunofluorescence tests on trypsinized cell lines growing as continuous cultures in a medium containing gamma globulins with specificity for the receptors. An attempt was made to determine the fate of the „surface receptor-serum globulin” complex by examining serial samples of cultures of stained positive cells for membrane immunofluorescence. There is partial evidence to suggest that observed fluctuations in the numbers of receptor-producing cells are partly due to active shedding of receptors from the cell surfaces.

Oocyte destruction and ovarian tumorigenesis after direct application of a chemical carcinogen (9:10‐dimethyl‐1:2‐benzanthrene) to the mouse ovary

Abstract: Ovaries of immature mice were painted with 9:10-dimethyl-1:2-benzanthracene (DMBA), and the effect of the carcinogen on the number of oocytes as well as on the subsequent development of ovarian tumours was studied. DMBA had an immediate effect on the small oocytes. Their number was rapidly reduced within the first 4 weeks after treatment, followed by a more gradual elimination, comparable to the elimination rate in the control ovaries. The growing and large oocytes were not primarily affected by the carcinogen; however, after the first 4 weeks post treatment their number was also decreased. This was considered to be secondary to the reduction in the number of small oocytes. No more oocytes remained in the ovaries of 6- to 9-month-old treated animals, an age at which considerable numbers were still present in the ovaries of control mice. Simultaneously with the elimination of oocytes there occurred a pathological development in the ovaries. There seemed to be a positive correlation between the rate of oocyte disappearance and the pathological development: the faster the oocytes disappeared the earlier the abnormal development began. This ultimately led to ovarian tumours, only seen in ovaries totally depleted of oocytes. It is concluded that DMBA applied to the ovaries acts directly on the small oocytes and destroys many of them. The neoplastic development is considered secondary to the reduction of the oocyte population and is apparently not caused by the carcinogen itself. This is supported by the fact that the development of ovarian tumours in mice is always preceded by a rapid disappearance of oocytes, regardless of whether the method of tumour induction in X-irradiation, intrasplenic transplantation of an ovary after gonadectomy, treatment with a chemical carcinogen, or genetic deletion of oocytes. Destruction des Oocytes et Tumorigenese Ovarienne apres Application Directe d'un Carcinogene Chimique (dimethyl-9,10 Benzo-1,2 Anthracene) sur l'ovaire de la Souris On a badigeonne les ovaires de souris immatures avec du dimethyl-benzanthracene (DMBA) et l'on a etudie l'effet de ce carcinogene sur le nombre des oocytes et sur la formation ulterieure de tumeurs ovariennes. Le DMBA a eu un effet immediat sur les petits oocytes. Leur nombre a rapidement diminue pendant les quatre semaines qui ont suivi le traitement; on a observe ensuite une elimination plus graduelle, comparable au taux d'elimination dans les ovaires temoins. Les grands oocytes en croissance n'ont pas ete, au debut, influences par le DMBA, mais apres les quatre premieres semaines consecutives au traitement leur nombre a egalement diminue. On a pense que ce phenomene etait secondaire a la diminution du nombre des petits oocytes. Six a neuf mois apres le traitement, il ne restait plus d'oocytes dans les ovaires des animaux, alors qu'il y en avait encore un nombre important dans les ovaires des souris temoins. En měme temps que l'elimination des oocytes on a observe une evolution pathologique dans les ovaires. Il semblait y avoir une correlation positive entre le taux de disparition des oocytes et l'evolution pathologique: plus les oocytes disparaissaient vite, plus l'evolution anormale commencait tǒt. Celle-ci aboutissait a des tumeurs ovariennes que l'on n'a constatees que dans les ovaires ou les oocytes avaient totalement disparu. On en conclut que le DMBA applique sur les ovaires agit directement sur les petits oocytes et en detruit la plupart. On estime que l'evolution neoplasique est secondaire a la diminution de la population d'oocytes et ne semble pas ětre causee par le carcinogene lui-měme. Cette hypothese est renforcee par le fait que la formation de tumeurs ovariennes chez la souris est toujours precedee par une disparition rapide des oocytes, quel que soit le mode d'induction de la tumeur: irradiation par les rayons X, transplantation intrasplenique d'un ovaire apres gonadectomie, traitement par un carcinogene chimique ou destruction genetique des oocytes.

Aflatoxin carcinogenesis: inhibition of liver cancer induction in hypophysectomized rats.

Abstract: Intact and hypophysectomized male rats were fed aflatoxin at a dose (4 μg/g in diet) sufficient to produce liver cancers in all of the 14 intact rats surviving 49 weeks Fourteen hypophysectomized rats also survived this latent period, but no liver tumors were induced although the animals received aflatoxin at a higher rate than the intact control animals However, tumors of extrahepatic tissues—particularly of the retro-orbital lacrimal glands—were induced in the hypophysectomized rats The selective blockade of hepatocarcinogenesis by hypophysectomy previously known with aminoazo dyes and fluorenamines is therefore extended to a new class of chemicals, the aflatoxins It is suggested that aflatoxins may require metabolic activation before becoming effective carcinogens for rat liver

Immunology of spontaneously arising rat mammary adenocarcinomas.

Abstract: The immunogenicities of nine spontaneously arising mammary adenocarcinomas were assessed following pre-treatment of syngeneic female rats with heavily irradiated tumour or excision of subcutaneously developing tumour grafts. One tumour (SP4) was moderately immunogenic and a weaker level of resistance could be evoked against two others. With these three examples, induced resistance was specific to the immunizing tumour and no instances of antigenic cross-reaction were detected. No resistance could be induced against the remaining tumours, even though challenge inocula of as few as 102 or 103 cells were tested in pre-irradiated hosts. There was no correlation in this series between tumour immunogenicity and latent period of development. Peritoneal exudate cells from rats immune to tumour Sp4 were shown to suppress the growth of this tumour in normal pre-irradiated recipients, but relatively large numbers of immune cells were necessary and their effectiveness was not consistent, factors which reflect the low immunogenicity of the tumour. These peritoneal cells had no influence on the growth of any other of the spontaneous rat mammary adenocarcinomas, further confirming the individuality of the TSTAs on tumour SP4.

Some factors affecting membrane immunofluorescence reactivity of burkitt lymphoma tissue culture cell lines

Abstract: Membrane immunofluorescence (MIF) reactivity of Epstein-Barr virus (EBV) carrying cultured Burkitt's lymphoma (BL) cells with BL sera shows certain fluctuations from time to time, even with standard reference sera and apparently constant tissue culture conditions. In order to determine the culture conditions that are critical for this variation and to establish the optimal situation favoring the production of highly reactive cells, we studied two established BL-derived cell lines under different conditions of initial cell number, cell density per unit volume or bottom unit area in the culture vessel, depth of the fluid phase, proportion of conditioned medium, etc. Although the two lines behaved somewhat differently, they showed better reactivity when maintained at a high concentration, as long as crowding did not entirely prevent cell multiplication. Under identical conditions of medium supply and depth of the fluid phase, more closely packed cells within a smaller bottom area of the culture vessel exhibited a better MIF reactivity than more dispersed cells. Under different conditions of growth, there was an inverse relationship between the rate of cell multiplication and the percentage of viable MIF-positive cells. Optimal conditions with steady, high-level reactivity have been worked out for both lines, but were different with regard to quantitative detail. Immunofluorescence de la Membrane des Cellules de Burkitt La reactivite a l'immunofluorescence de la membrane (IFM) des cultures de cellules du lymphome de Burkitt (LB) contenant des virus d'Epstein-Barr (EB), en presence de serum de LB, montre que crrtaines fluctuations se produisent de temps a autre, meme aver des serums de reference standard et dans des conditions appareniment constantes de culture tissdaire. Afin de definir les conditions de culture qui sont critiques pour ces variations et de determiner la situation optimafe qui favorisr la production de cellules forterrrent reactives, deux lignees cellufaires etahlies, derivees du L B, ont ete etudiees dans differentes conditions de nombre initial de cellules, de densite callulaire par volume unitaire ou sur la surface du fond du recipient de culture, de hauteur de la phase liquide, de proportion de milieu conditionne, etc. Bien que Ies deux lignees se soient comportees assez differemment, elles ont fait preuve d'une meilleure reactivite lorsqu'elles sont restees fortement concentrees, tant que I'agglomeration n'a pas empeche totalement la multiplication cellulaire. Dans des conditions identiqrres de preparation du milieu et de hauteur de la phase liquide, des cellules plus etroitement tassees sur une plus petite surface du fond du recipient de culture ont fait preuve d'une meilleure reactivite a l'IFM que des cellules plus dispersees. Dans des conditions de proliferation differentes, il existe une relation inverse entre le rythme de multiplication cellulaire et le pourcentage de cellules viables reagissant positivement a l'IFM. Des conditions optimales, avec une reactivite constante et elevee, ont ete obtenues pour les deux lignees, mais elles etaient differentes dans le detail au point de vue quantitatif.

The lipids of normal diploid (WI‐38) and sv40‐transformed human cells

Abstract: The lipids of human diploid cell strain WI-38 and of a line derived from it by SV40 transformation have been studied. The lipid composition and fatty acid spectra of both resemble those reported for other tissue culture cells and human tissues. The lipids of the normal and transformed cells are similar except that there are decreases in the content of phospholipids and of arachidonic acid in the transformed cells. The ability of the cells to synthesize lipids was assayed in a „resting” cell system. Both cell types were able to incorporate labelled acetate into their component neutral lipids and phospholipids, but the rates observed were lower than that required for cell growth. Elevation in the rate of cholesterol synthesis was observed in the transformed cells. The data are discussed in relation to other reports in the literature on lipids and neoplasms.

Studies on Murine Sarcoma Virus: antigenic characterization of Murine Sarcoma Virus induced tumor cells.

Abstract: The antigenicity of Murine Sarcoma Virus (MSV) transformed mouse, rat and hamster cells has been studied by several techniques designed to detect surface antigens. Antigens were readily demonstrated on neoplastic mouse and rat cells. There was complete cross-reactivity between cells induced by the Harvey (MSV-H) or Moloney (MSV-M) strains of MSV and by Moloney Leukemia Virus (MLV). No evidence of a distinct or separate „MSV-non-MLV” antigen could be obtained by cross absorptions of MSV/MLV antisera and cells, or by tests designed to break MLV tolerance. Such results favor the view that MSV and MLV are closely related entities. With the 8303 line of MSV-M transformed hamster cells serological tests revealed little if any reactivity with strongly positive mouse anti-MSV/MLV antisera. In one test, however, immunization of mice with these cells gave some degree of protection against syngeneic MSV sarcoma.

The occurrence of tumours in F1, F2, and F3 descendants of pregnant mice injected with 7,12-dimethylbenz(a)anthracene.

Abstract: Pregnant MA mice were injected intraperitoneally with 400 micrograms of 7,12-Dimethylbenz(a)anthracene in olive oil during the last stages of pregnancy. The genetic design of the experiment included the follow-up of the descendants for three consecutive generations and the cross-mating between the F1 descendants of treated mothers with descendants of untreated controls. An increased incidence of tumours was observed in the female F1 and F2 descendants and the female F2f descendants (cross-mating of F1 female descendants from a treated mother and male descendants from an untreated control). The tumours were observed in animals dying at a late age which approximately corresponded to the age when spontaneous tumours were observed in control animals. The most frequent types of tumours were the same as observed in the control animals, together with a handful of rarely observed tumours. In the F1 and F2 male descendants the tumour incidence was higher than in control animals but the difference was statistically not significant (binomial test). No significant increase in tumour incidence was observed in the F3 and F2m descendants (cross-mating of F1 male descendants from treated mothers with female descendants from untreated controls). While the mice of F1 generation were directly exposed to small amounts of the chemical carcinogen via the mothers' milk and excreta, and possibly through the placenta, it seems unlikely that DMBA as such was transmitted to the F2 descendants.

Ribonuclease-susceptible charged groups at the surface of ehrlich ascites tumour cells.

Abstract: The electrophoretic mobilities of Ehrlich ascites tumour cells are significantly reduced following treatment with bovine pancreatic ribonuclease. This loss of net surface negativity has been intensively investigated, and has been shown to be due to the enzymatic activity of the ribonuclease, as distinct from non-specific adsorption of this basic protein. The experimental data are consistent with the concept that RNA is a structural component of the cell periphery. Groupements Charges a la Surface des Cellules Les mobilites electrophoretiques des cellules tumorales des ascites d'Ehrlich diminuent notablement apres un traitement a la ribonuclease pancreatique bovine. Cette baisse de la charge negative nette de la surface a fait l'objet de recherches approfondies qui ont montre qu'elle est due a l'activite enzymatique de la ribonuclease, qu'il ne faut pas confondre avec l'adsorption non specifique de cette proteine de base. Les donnees experimentales confirment la theorie selon laquelle l'ARN est un element structurel de la peripherie cellulaire.

Effective antiviral therapy of two murine leukemias with an interferon-inducing synthetic carboxylate copolymer

Abstract: A synthetic anionic copolymer was employed to induce interferon in vivo. The ability of the induced interferon to protect mice inoculated with lethal doses of Friend or Rauscher leukemia virus was examined. Four parameters were employed to assess the antiviral activity of the induced interferon; inhibition of splenomegaly; viral-induced spleen foci; viral replication; and increase in survival time. Daily treatment with a low dose of copolymer was more effective than a single treatment with a high dose. A higher percentage of treated mice was completely protected from a lethal dose of Friend or Rauscher leukemia virus when compared with the controls. A relationship occurred between the effect of copolymer-induced interferon and the infecting dose of leukemia virus. Protection developed only when treatment was initiated prior to virus challenge.

Immunology of 2-acetylaminofluorene-induced rat mammary adenocarcinomas.

Abstract: The immunogenicities of 11 rat mammary adenocarcinomas induced with 2-acetylaminofluorene (AAF) have been studied. Attempts were made to induce tumour-specific immunity in syngeneic female rats, following treatment with heavily irradiated tumour or surgical removal of developing subcutaneous tumour grafts. Resistance was evoked against only two tumours and this was extremely weak. With the others, no resistance against the immunizing tumour could be demonstrated, even though the tests were made highly sensitive by the use of inocula of as few as 103 cells in pre-irradiated hosts. Sera from rats immunized against six of the tumours were tested against viable cells of the immunizing tumour in an immunofluorescence test. In no case did tumour antisera give a positive reaction with the tumour cells, although allogeneic isoantisera consistently gave strongly positive reactions. The results are interpreted as showing that AAF-induced rat mammary adenocarcinomas are deficient in tumour-specific transplantation antigens. Immunologie de l'Adenocarcinome Mammaire du rat Provoque par l'Acetylamino-2 Fluorene On a etudie l'immunogenicite de 11 adenocarcinomes mammaires du rat provoques par l'acetylamino-2 fluorene (AAF). On a essaye de provoquer une immunite tumori-specifique chez des rates syngeneiques apres traitement par une tumeur fortement irradiee ou exerese chirurgicale de greffons sous-cutanes. Une resistance n'a ete suscitee que vis-a-vis de deux tumeurs; elle etait d'ailleurs extrěmement faible. Avec les autres, il a ete impossible de mettre en evidence une resistance contre la tumeur immunisante, bien que les epreuves aient ete rendues hautement sensibles par l'emploi d'inoculums tres faibles (103 cellules) chez les hǒtes pre-irradies. Les serums de rates immunisees contre six des tumeurs ont ete mis en presence de cellules viables de la tumeur immunisante dans une epreuve d'immunofluorescence. Les serums anti-tumeurs n'ont en aucun cas donne une reaction positive avec les cellules tumorales, alors que des iso-antiserums allogeneiques ont regulierement donne des reactions fortement positives. Les auteurs en concluent que l'adenocarcinome mammaire du rat induit par l'AAF manque d'antigenes de transplantation specifiques.

The sensitivity of a yoshida sarcoma to methylene dimethane sulphonate

Abstract: The action of methylene dimethane sulphonate on the growth rate of the Yoshida sarcoma in rats is described. 1) The tumour, even when established for 12 to 14 days, is unusually sensitive to this alkylating agent and approximately 90% complete cures can be obtained. 2) About 10% of the treated tumours recur and, on retransplantation to new animals, show a marked resistance to the drug. The growth curves of the original and resistant lines, both unilaterally and bilaterally transplanted, following treatment by the drug, are described. 3) Animals in which the original tumour had completely regressed failed to accept a reimplantation of the original or of a resistant tumour. Comparable doses of myleran failed to produce complete regression of the tumour when administered 10 days after transplantation and the level of inhibition was much less than with methylene dimethane sulphonate. Cross-resistance with triethylene melamine was observed. Sensibilite d'un Sarcome de Yoshida au dimethanosulfonate de Methylene L'article decrit l'action du dimethanosulfonate de methylene sur le taux de croissance du sarcome de Yoshida chez des rats. 1) La tumeur, měme quand elle est installee depuis 12 a 14 jours, est exceptionnellement sensible a cet alcoylant et l'on peut obtenir environ 90% de guerisons completes. 2) Environ 10% des tumeurs traitees recidivent; lorsqu'elles sont regreffees a des animaux neufs, elles presentent une resistance marquee au medicament. On analyse les courbes de croissance des tumeurs initiales et des tumeurs resistantes, transplantees unilateralement et bilateralement, apres traitement par ce produit. 3) Les animaux chez lesquels la tumeur initiale avait completement regresse n'ont accepte ni une reimplantation de la tumeur initiale ni celle d'une tumeur resistante. Des doses comparables de myleran n'ont pas permis d'obtenir une regression complete lorsqu'elles etaient administrees 10 jours apres la greffe, et le degre d'inhibition etait bien moindre qu'avec le dimethanosulfonate de methylene. On a observe une resistance croisee avec le triethylene melamine.