Showing papers in "International Journal of Cancer in 1983"

Production of a mouse monoclonal antibody reactive with a human nuclear antigen associated with cell proliferation

Abstract: The production of a mouse monoclonal antibody, Ki-67, is described. The Ki-67 antibody recognized a nuclear antigen present in proliferating cells, but absent in resting cells. Immunostainings with Ki-67 revealed nuclear reactivity in cells of germinal centres of cortical follicles, cortical thymocytes, neck cells of gastrointestinal mucosa, undifferentiated spermatogonia and cells of a number of human cell lines. The Ki-67 antibody did not react with cells known to be in a resting stage, such as lymphocytes, monocytes, parietal cells and Paneth's cells of gastrointestinal mucosa, hepatocytes, renal cells, mature sperm cells, brain cells, etc. Expression of the antigen recognized by Ki-67 could be induced in peripheral blood lymphocytes after stimulation with phytohaemagglutinin, whereas it disappeared from HL-60 cells stimulated with phorbol esters to differentiate into mature macrophages in a resting stage. These findings suggest that Ki-67 is directed against a nuclear antigen associated with cell proliferation. A first series of immunostainings of tumour biopsies indicated that Ki-67 may be a potent tool for easy and quick evaluation of the proportion of proliferating cells in a tumour.

Human papillomavirus type‐16‐related DNA in genital Bowen's disease and in bowenoid papulosis

Abstract: 32P-labelled DNA of HPV 16 which has been isolated and molecularly cloned from a cervical carcinoma (Durst et al, 1983) was used to screen the cellular DNAs obtained from 20 different biopsies of Morbus Bowen or Bowenoid papulosis, respectively, by Southern blot analysis Under conditions of differing stringency for the hybridization, HPV 16 DNA or related sequences were identified in 6 out of 10 cases of Morbus Bowen (4 out of 5 from a genital localization) and in 8 out of 10 biopsies from Bowenoid papulosis One additional case of the latter disease contained DNA sequences of an HPV type not yet classified There is evidence for the presence of another HPV DNA in two of the HPV-16-positive tumors A large number of normal genital tissue samples were negative for HPV DNA

Diet and colorectal cancer: A case-control study in Greece

Abstract: A case-control study probing the role of diet on the incidence of colorectal cancer was undertaken in Athens, Greece, in a population characterized by ethnic homogeneity but substantial heterogeneity with respect to dietary habits. The case series consisted of 100 consecutive patients with histologically confirmed colorectal cancer admitted to two large hospitals of Athens during a 16-month period; the control series consisted of orthopaedic patients, admitted to the same hospitals during the same time period, individually matched to the index cases by age and sex. Dietary histories concerning the frequency of consumption (per month or per week) of about 80 food items were obtained by the same interviewer. Cases reported significantly less frequent consumption of vegetables (particularly beets, spinach, lettuce and cabbage) and, independently, significantly more frequent consumption of meat (notably lamb and beef). Between the two extremes (high-vegetable, low-meat diet versus high-meat, low-vegetable diet) a risk ratio of about 8 appears to exist, sufficient (in size and direction) to explain a substantial part of the international variation in the incidence of colorectal cancer. Significant associations were not found with beer or other alcoholic beverages, and significant interactions were not noted with respect to age, sex and anatomic localization (colon vs. rectum).

Dietary habits and lung cancer risk.

Abstract: A previously reported negative association between a high index of dietary vitamin A and lung cancer incidence was confirmed in an extended follow-up, covering 11 1/2 years, of 13,785 men and 2,928 women, Responses to a postal questionnaire provided the dietary information. Relationships between the major dietary items and lung cancer were explored for various diagnostic subsets of the 168 lung cancer cases diagnosed among the study subjects. Analyses were stratified for sex, age, residence characteristics, cigarette smoking and, at times, socioeconomic group. Although the data do not permit a firm interpretation in terms of risk enhancement by a marginal retinoid deficiency, we found that the apparent protection afforded by higher intakes of vitamin A or its provitamins was particularly strong for lung cancer appearing as squamous-cell carcinoma and among those with higher alcohol intakes. The individual food items which showed the strongest negative association with lung cancer were carrots and milk. These two items made a major contribution to the vitamin A index and its variation among the respondents.

Food items and food groups as risk factors in a case-control study of diet and colo-rectal cancer.

Abstract: The contributions of food items and food groups as risk factors in a previously reported case-control study of diet and colo-rectal cancer have been analyzed. The study included 348 patients with colon cancer, 194 with rectal cancer, 542 neighbourhood controls individually matched to the cases on the basis of age and sex and a second control series of 535 surgical hospital controls frequency matched to the cases. For colon cancer, as in the previous analysis, the major risk factor was saturated fat, individual food items or groups failing to make a significant contribution to the risk. In particular there was no protective effect of dietary fibre and, for cruciferous vegetables, only a minor protective effect in females. No individual cruciferous vegetable made an important contribution to this effect. For rectal cancer, on the other hand, a significant effect of saturated fat, independent of other food items or groups, was only found for females in the highest consumption category. For males, consumption of eggs, beef and veal significantly increased risk but not consumption of pork, while for females, there was a non-significant increase in risk with consumption of eggs, no increased risk with consumption of beef or veal and a significantly increased risk with consumption of pork. There was no protective effect of dietary fibre or of cruciferous vegetables for rectal cancer, but in females, there was a significantly increased risk for consumption of beer, though this was somewhat reduced when controlled for consumption of saturated fat. There was no indication of an effect of alcohol in either sex or of beer in males. Thus, these results confirm the previous report in showing a significant effect of saturated fat in increasing risk of colon cancer but suggest a contribution of meats to risk of rectal cancer.

Age at any birth and breast cancer risk

Abstract: In an effort to assess the relative importance of age at first birth, age at subsequent births, and total parity to the occurrence of breast cancer, reproductive data from 4,225 women with breast cancer and 12,307 hospitalized women without breast cancer were analyzed by a multiple logistic regression model. Age at first birth was confirmed to be the most important reproductive risk indicator; it was associated with a 3.5 % increase of relative risk for every year of increase in age at first birth (the 95 % confidence interval of this estimate was 2.3 to 4.7 % increase per year). However, age at any birth after the first was also an independent and statistically significant risk indicator; it was associated with a 0.9% increase of relative risk for every year of increase in age at any (and every) birth (the 95 % confidence interval of this estimate was 0.4 to 1.5 % increase per year). There is evidence that the age of approximately 35 years represents for every birth a critical point; before this age any full-term pregnancy confers some degree of protection; after this age any full-term pregnancy appears to be associated with increase in breast cancer risk. The effect of parity is determined by the age of occurrence of the component pregnancies. While most pregnancies occur under the age of 35, the distribution varies from population to population, and this may account for the differences between populations in whether or not a protective effect is seen for births after the first, and if it is seen, its extent.

Quantitating the synergistic effect of smoking and alcohol consumption with the micronucleus test on human buccal mucosa cells.

Abstract: The micronucleus test was applied to exfoliated cells of the buccal mucosa of four population groups: (A) non-smokers and non-drinkers of alcoholic beverages, (B) non-smokers but alcohol drinkers, (C) smokers but non-drinkers, and (D) smokers and drinkers. An elevated frequency of micronucleated buccal mucosa cells was observed only in group D (smokers and alcohol drinkers). When group D was subdivided according to the number of cigarettes smoked, the frequency of micronucleated buccal cells and the average number of micronuclei per cell appeared to depend upon cigarette consumption. An approximately eight-fold increase of micronucleated mucosa cells was seen among alcohol drinkers who smoked three or more packs of cigarettes per day, and an approximately 4.2-fold elevation was observed when one to two packs were consumed. Neither smoking alone of up to and over 60 cigarettes per day nor ethanol drinking alone of up to 1.21 per day led to a detectable elevation of micronucleated buccal mucosa cells. Whether the strong synergistic effect between smoking and alcohol consumption, as seen by the frequency of micronucleated buccal mucosa cells, is related to their synergistic effect in the induction of oral cancers is an intriguing but open question.

Human T‐cell leukemia virus type I: Induction of syncytia and inhibition by patients' sera

Abstract: HTLV-producing T-cell lines induce cell fusion when co-cultivated with a wide variety of indicator cells, suggesting that HTLV envelope antigens interact with the membranes of many cell types. Serum antibodies from adult T-cell lymphoma-leukemia (ATL) patients inhibited the formation of syncytia, and sera from British, American and Japanese ATL patients inhibited cell fusion induced by American and Japanese HTLV isolates equally well. No serological cross-inhibition of syncytium induction was found between HTLV and bovine leukosis virus, Moloney murine leukemia virus and simian sarcoma-associated virus. A simple syncytium inhibition test in microtiter plates has been developed to provide a rapid screen for antibodies presumed to be specific to HTLV envelope glycoproteins.

Differential reactivity of monoclonal antibodies with human colon adenocarcinomas and adenomas.

Abstract: Monoclonal antibodies have been generated using membrane-enriched extracts of human metastatic mammary carcinoma lesions (Colcher et al., 1981), some of which demonstrated binding to the surface of human colon carcinoma cell lines. We report here an analysis of the reactivity of three of these monoclonal antibodies with formalin fixed tissue sections of human colon adenocarcinomas and adenomas. The three monoclonals employed were B72.3, which is reactive with a 220-400 kdal high molecular weight glycoprotein complex; B6.2, reactive with a 90 kdal glycoprotein, and B1.1, which is reactive with the 180 kdal glycoprotein CEA. B1.1 was least selective in its reactivity to colon carcinoma versus adenoma lesions. When 10 micrograms/ml of purified B1.1 IgG were used per slide, 94% (15 of 16) of carcinomas and 83% (15 of 18) adenomas showed reactivity. Monoclonal B72.3 demonstrated the most selective reactivity for carcinomas. Eighty-two percent (14 of 17) of carcinomas were positive while none of 18 adenomas examined showed reactivity with more than a few percent of adenoma cells positive. When a low concentration of purified B72.3 immunoglobulin was used per slide, 8 of 16 carcinomas and none of 46 adenomas or normal colon epithelium samples scored positive. Monoclonal B72.3 also reacted with cells in areas of "atypia" within adenomas. The reactivity of monoclonal B6.2 was intermediate as compared to B1.1 and B72.3 in its selectivity of reactivity for carcinoma cells. A heterogeneity in the populations of tumor cells showing reactivity with the three monoclonals was observed within many of the tumor masses. Both colon adenocarcinomas and adenomas can now be placed in several distinct groups based on their expression of antigens reactive with the three monoclonal antibodies employed.

Selection by trypsin of two sublines of rat colon cancer cells forming progressive or regressive tumors

Abstract: From an established cell culture line obtained from a chemically-induced rat colon carcinoma, two sublines have been selected and isolated according to their susceptibility to trypsin-mediated detachment from plastic surfaces. Subline TR, the most resistant to the detaching effect of trypsin, gave progressive tumors in most of the syngeneic rats in which it was inoculated. Subline TS, which was easily detached by trypsin, also gave tumors in the syngeneic rats, but all these tumors disappeared within 3 or 4 weeks. Both sublines gave progressive tumors when injected into nude mice. This suggests that the parent cell line is heterogeneous and contains cell variants differing in their susceptibility to trypsin-mediated detachment from substrate and their sensitivity to host factors leading to acceptance or rejection of the inoculated tumor cells.

Detection of serum antibodies to adult T‐cell leukemia virus in non‐human primates and in people from Africa

Abstract: The distribution of serum antibodies to adult T-cell leukemia virus (ATLV) was examined as a marker for virus infection among non-human primates as well as people from Africa and Germany. The virus is present in Africa in certain primate species including man. Altogether, 468 sera from 27 monkey species were examined. Only African green monkeys, less frequently also chimpanzees and crab-eating monkeys, were found to be infected. About 1-2% of people from Kenya have antibodies, while ATLV-antibodies may be present in well below 0.1% of the German population.

Sucrase-isomaltase: A marker of foetal and malignant epithelial cells of the human colon

Abstract: The presence of sucrase-isomaltase (SI), a glycoprotein hydrolase normally restricted to the brush border membrane of the enterocytes of the small intestine, was investigated in tumours which developed in nude mice inoculated with six human colon carcinoma cell lines (HT-29, Caco-2, HRT-18, HCT-8R, SW-480, and CO-115). Foetal and normal adult human small intestines and colons were used as controls. SI was studied by (1) immunofluorescence with rabbit antibodies raised against purified human small intestine SI; (2) polyacrylamide gel electrophoresis and immunoblotting; and (3) determination of the enzyme activity. SI was antigenically present, and enzymatically active, in all the tumours derived from Caco-2 and HT-29 cells. The presence of the enzyme was associated with that of typical brush borders at transmission electron microscopy examination. SI was absent from the tumours developed with the other four cell lines, as well as from the normal adult colon mucosa. SI was also present and active in the colons of mid-gestation foetuses, ranging in ages between 20 and 28 weeks; it was absent from the colons of late-gestation foetuses. The presence of SI in tumours derived from two cell lines suggests that this enzyme is a marker, so far unsuspected, of certain human colon cancers, and that the differentiation pattern of these particular cancers closely resembles that of the foetal colon.

Formation and growth of multicellular spheroids of human origin.

Abstract: Different types of human cells which normally grow as monolayers or suspension cultures were tested for their capacity to form and grow as spheroids. Sixteen out of the 27 tested tumour cell lines formed spheroids. Nearly all of these spheroids also grew. With only two exceptions the doubling times were longer when the tumour cells grew as spheroids than when they grew in conventional mass culture. Eleven out of 13 tested human non-tumour cells formed small spheroids but of these only the spheroids of lymphoid origin could grow. These lymphoid cells grew faster when aggregated to spheroids than when in single-cell suspension culture. None of the other non-tumour cells, which normally grew as monolayers, could grow as spheroids. The normally monolayer-cultured tumour cells formed symmetrical spheroids with smooth surfaces while the normally suspension-cultured cells formed irregular spheroids with rough surfaces. All large spheroids had a necrotic centre surrounded by a shell of viable cells. The thickness of the viable cell layer varied depending on cell type. The shape and organization of cells within the spheroids also varied largely. The results show that many types of human cells can be cultured as spheroids and that a wide spectrum of morphological appearances and growth rates can be obtained.

Biological characterization and clinical applications of a monoclonal antibody recognizing an antigen restricted to neuroectodermal tissues.

Abstract: The monoclonal antibody UJ13A was raised following immunization of mice with human foetal brain and subsequent somatic cell hyridization of spleen cells with the mouse myeloma cell line P3-X63-AG8-653. The antibody is of the IgG1 subclass and has been shown by indirect immunofluorescence studies on normal foetal, paediatric and adult tissues to selectively bind to most tissues of neuroectodermal origin. Many tumours of neural origin also express the UJ13A antigen and the reagent can be used to distinguish primary intracranial neural tumours from secondary carcinomas and lymphomas. UJ13A is also useful as one of a panel of reagents employed for the identification of metastatic spread of neuroblastoma cells to bone marrow and cerebrospinal fluid. Knowledge of the full spectrum of normal and malignant tissues binding UJ13A suggests that the antibody may have a role in the radioimmunolocalization of neuronal tumours such as neuroblastoma.

Childhood cancer incidence: Geographical and temporal variations

Abstract: Data from the first four volumes of Cancer Incidence in Five Continents (CI-5) and from the first 5 years of the US Surveillance, Epidemiology and End Results (SEER) program were analyzed for evidence of geographical and temporal variations in the incidence of selected childhood tumors. Only lymphoid leukemia and glial neoplasms are common enough for the observed differences between US registries to be distinguished from sampling variation. Internationally, kidney and eye tumors and leukemia show less geographical variation than do lymphomas and brain tumors, but for none of the tumors examined is the incidence constant. Wilms' tumor rates among Japanese, Singapore Chinese and Indians (Bombay) are approximately 60% of the rates in North America and Britain, whereas in Scandinavia the rates are up to 30% higher. This lessens the status of Wilms' tumor as an "index tumor" of childhood. Areas or countries with especially high or low rates of other tumors are identified. Rates for glial neoplasms (SEER data) and Hodgkin's disease (CI-5) are increasing with time in the US, while brain tumors are being diagnosed more frequently worldwide. However, the results for brain tumors may largely reflect changes in pathology diagnosis or reporting practices, and those for Hodgkin's disease may reflect improvements in case ascertainment. Otherwise, there is a remarkable stability in the incidence of selected childhood cancers over time.

Effects of several species of human leukocyte interferon on cytotoxic activity of NK cells and monocytes

Abstract: Ten species of purified human leukocyte interferon were tested for their ability to modulate the cytolytic activity of natural killer (NK) cells and the cytolytic and cytostatic activities of monocytes. The interferon species were tested at several antiviral titers and examined for quantitative differences in their ability to modulate immunological function. At the higher doses of interferon (i.e., greater than 500 units) all of the interferon species demonstrated significant augmentation of cytolysis and cytostasis. However, when low levels (i.e., 10-50 units) of interferon were employed, appreciable differences between the various interferon species were seen. A similar pattern of relative potency among the various species of pure leukocyte interferon was seen for augmentation of cytolysis by monocytes and NK cells. In contrast, a different pattern of relative potency was observed for augmentation of cytostasis. These results demonstrated substantial quantitative differences (as much as 100-fold) in the ability of the various species of human leukocyte interferon to induce significant levels of augmentation of these cell-mediated functions. Such results should have significant impact in choosing a specific interferon species for appropriate clinical trials.

MHC imbalance and metastatic spread in Lewis lung carcinoma clones

Abstract: Imbalance in the Kb and Db region encoded molecules is observed in Lewis lung carcinoma clones. The uncloned metastatic population and the D122 high-metastatic clone show no expression of H-2Kb products, while the nonmetastatic A9 clone expresses Kb products. Twenty-nine new subclones of 3LL and A9 were analyzed for D-end and K-end membrane expression, primary growth rate and metastatic spread. We show that the imbalance in H-2Kb to H-2Db is correlated with metastatic properties of a given clone, but local tumor growth is not. A "low Kb/low Db" phenotype is nonmetastatic as is a "high Kb/high Db" phenotype; a "low Kb/high Db" is highly metastatic and a "medium Kb/high Db" is moderately metastatic. We find support for this notion of imbalance in experiments on MHC modulation by interferon and retinoic acid. Interferon increases both Kb and Db expression of A9 and D122 clones yet the net increase of Db was greater than Kb. This was associated with an increase in metastasis formation. Retinoic acid increases the expression of the Db gene product on the nonmetastatic A9, clone, without apparent changes in Kb expression. This treatment shifts the A9 to a high-metastatic phenotype. The significance of this imbalance to the tumor--host relationship is discussed.

HLA and nasopharyngeal carcinoma in Chinese--a further study.

Abstract: This paper reports the results of 8 years of HLA typing of newly diagnosed nasopharyngeal carcinoma (NPC) patients among Chinese in Singapore, and compares the HLA profile of NPC cases with that of the normal Chinese population. Earlier results had indicated an association with A2 and BW46, but the heterogeneity of the earlier case series, which contained a proportion of long-term survivors, obscured other aspects of the relationship. In this paper, the risk associated with A2 (relative risk = 1.5) and BW46 (relative risk = 1.9) is confirmed, showing a much higher relative risk in the presence of both antigens, and a risk is also demonstrated for B17 (relative risk = 2.1). Relative risks of less than unity are observed for AII (relative risk = 0.5) and B13 (relative risk = 0.5). Data from family studies indicate the importance of the haplotype A2.BW46 in determining risk for NPC.

Prospective study of "early" gastric cancer.

Abstract: In order to elucidate the natural history of early gastric cancer, we followed up non-concurrently certain patients who had been diagnosed endoscopically as having "early' gastric cancer and who had histological evidence of cancer by biopsy, but on whom surgical resection had been delayed or had not been conducted. At the Center for Adult Diseases, Osaka, 56 cases were eligible for this study. Out of these, 27 cases progressed to advanced cancer and 16 remained at the early stage during the follow-up period (6-88 months, mean: 29 months). The remaining 13 cases had had neither surgical resection nor examinations. The survivorship function for the duration of "early" gastric cancer was estimated by the life-table method of Kaplan and Meier. The median of the duration was estimated as 37 months. The 5-year survival rate of the 34 cases on whom surgical resection had not been conducted was estimated as 64.5%. These results suggest that early diagnosis and early treatment may lead to a reduction of mortality from stomach cancer.

Preparation and properties of a drug-carrier-antibody conjugate showing selective antibody-directed cytotoxicity in vitro.

Abstract: The preparation and properties of a drug-carrier-antibody preparation are reported. The antifolate chemotherapeutic agent methotrexate was covalently coupled to human serum albumin as a carrier. The carrier-drug preparation was then chemically linked to a monoclonal antibody, raised originally against a human osteogenic sarcoma cell line, 791T, in a manner permitting retention of antibody-binding activity. The cytotoxic properties of the conjugate were tested in vitro in comparison with carrier-methotrexate and free methotrexate against a panel of tumour cell lines containing both antigenically cross-reactive cell lines and cell lines having low antigenic cross-reactivity with the monoclonal antibody. The cytotoxicity tests demonstrated that coupling of methotrexate to carrier caused a loss of some drug activity but that coupling of the antibody to the carrier-drug preparation permitted full expression of drug cytotoxicity against antibody-reactive cell lines. It was further demonstrated that the conjugate was selective in its action and was preferentially cytotoxic towards antibody-reactive cell types. The cytotoxicity against antibody-reactive cell lines was shown by competitive inhibition by free antibody to be entirely dependent on antibody binding. A clonogenic assay showed that the conjugate was capable of killing greater than 99% of 791T target cells. These results indicate that a drug-carrier antibody conjugate can be synthesized which has all the in vitro properties theoretically necessary for a successful antibody-targeted cytotoxic agent.

Cytogenetic studies on an epithelial cell line derived from poorly differentiated nasopharyngeal carcinoma

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

Mortality patterns among embalmers.

Abstract: In view of recent findings of nasal cancer in rats exposed to formaldehyde vapors, we investigated the proportionate mortality experience of embalmers licensed to practice in New York State. Mortality was significantly elevated for cancers of the skin and colon and for arteriosclerotic heart disease, whereas significant deficits were seen in mortality from respiratory diseases and accidents. Respiratory cancer mortality was not excessive and no deaths were attributed to nasal cancer. Mortality was significantly elevated for cancers of the skin, kidney, and brain among those licensed only as embalmers, whereas mortality patterns were unremarkable among those licensed also as funeral directors (and presumably less exposed to formaldehyde) These preliminary results indicate the need for occupational cohort studies to clarify the carcinogenic potential of formaldehyde.

Oesophageal cancer in non-smoking drinkers and in non-drinking smokers

Abstract: Alcohol and tobacco are the two major risk factors for cancer of the oesophagus; they usually combine their action. The action of each factor is studied here in non-users of the other. Among non-drinking male smokers the relative risk is higher than 5; among non-smoking drinkers, risks are much higher and increase with average daily intake; this effect is also observed in females, with risk values of the same order of magnitude as for males. Thus the high sex ratio of oesophageal cancer in France can be entirely ascribed to different drinking levels in females and in males.

A comparison of epstein‐barr virus‐specific T‐cell immunity in malaria‐endemic and ‐nonendemic regions of Papua New Guinea

Abstract: Epstein-Barr virus genome-positive Burkitt's lymphoma is endemic in Africa and Papua New Guinea and in both countries the tumour is restricted to regions with holoendemic malaria. The present work has compared groups of healthy indigenous individuals living in malarious and non-malarious regions of Papua New Guinea for Epstein-Barr virus-specific T-cell-mediated immunity using the in vitro regression assay. Residents of the malarious region (55 tested), when compared with either residents of the non-malarious area (35 tested) or Caucasian controls (27 tested) showed a significant (p less than 0.0001) impairment of virus-specific T-cell immunity but no obvious disturbance (p greater than 0.05) of anti-viral antibody titres. These results may be important in explaining the postulated role of malarial infection as a co-factor in the pathogenesis of Burkitt's lymphoma.

Forskolin: A potential antimetastatic agent

Abstract: Forskolin, a diterpene from the roots of an Indian plant, Coleus forskohlii, is a potent platelet aggregation inhibitor and has been examined for its effects on (a) tumor-induced human platelet aggregation and (b) pulmonary tumor colonization in mice. These studies employed a subline of B16 murine melanoma, B16-F10 (highly metastatic to lungs). Forskolin (2 microM) strongly inhibits the melanoma cell-induced human platelet aggregation. A single dose of forskolin (82 micrograms/mouse) administered intraperitoneally 30 or 60 min prior to tail vein injection of cultured B16-F10 cells (2 or 3 X 10(5) cells/mouse) reduced tumor colonization in the lungs by more than 70%. Similar results were obtained in three separate experiments. These findings raise the possibility that forskolin could prove of value in the clinic for the prevention of cancer metastasis.

Infection and transformation of fresh human umbilical cord blood cells by multiple sources of human T-cell leukemia-lymphoma virus (HTLV).

Abstract: Human T-cell leukemia-lymphoma virus (HTLV) was first isolated from sporadic patients with adult T-cell malignancies in the United States and subsequently from T-lymphocytes established in culture from additional T-cell leukemia-lymphoma patients living in different geographical areas of the world. Co-cultivation of normal umbilical cord blood with lethally irradiated, HTLV-positive lymphocytes established in culture from many of these patients resulted in the productive infection of the cord blood T-lymphocytes which grew in suspension culture in the absence of exogenous TCGF. These transformed cord blood cells have morphological and cytochemical properties similar to HTLV-positive fresh and cultured tumor T-cells and are distinguishable from virus donor cells by HLA haplotype and chromosomal markers. These cells express HTLV proteins, release type-C virus particles and contain surface receptors for TCGF. These results demonstrate that HTLV isolated from T-cell leukemic donors from different parts of the world can productively infect and transform fresh human cord blood T-lymphocytes, and that the transformed cells share many similarities with fresh or cultured leukemic cells.

A pigmentation-associated, differentiation antigen of human melanoma defined by a precipitating antibody in human serum.

Abstract: Antibodies in the serum of melanoma patient AU precipitate an antigen from 125I-labelled extracts of cultured autologous melanoma cells. The antigen, which is probably not a cell surface component, is present in other pigmented melanomas but not in non-pigmented melanomas or other tumor cell types, and the amount of antigen is correlated with the degree of pigmentation. These conclusions are based on absorption experiments with 11 pigmented melanomas, 8 non-pigmented melanomas, 3 astrocytomas, 12 carcinomas of various histological types, 1 leukemia, 2 EB-virus-transformed B lymphocyte lines, and human erythrocytes. The antigen was also detected in cultured human melanocytes. It has a molecular weight of 70,000, an isoelectric point of pH 5.3, and it binds to concanavalin A-Sepharose. Ninety-six sera from other melanoma patients were examined and none of them precipitated this antigen. As described previously, the serum from patient AU also has antibodies to a unique (Class 1) tumor antigen found only on AU melanoma cells. The pigmentation-associated, differentiation antigen and the unique antigen are clearly different in their distribution, but some relationship between these unusual antibody responses is possible.

Natural infection in non‐human primates with adult T‐cell leukemia virus or a closely related agent

Abstract: A total of 703 sera from 10 species of monkeys were examined for the presence of antibodies to adult T-cell leukemia (ATL)-associated antigens (ATLA). ATLA represent core protein(s) of ATL virus (ATLV) and ATLV-determined polypeptides. Anti-ATLA antibodies were found in all seven macaque species tested but not in three non-macaque species. The frequencies of seropositive macaques ranged from 10 to 50%. In three macaque species (Japanese monkeys, rhesus monkeys, and crab-eating monkeys) in which sufficient numbers of animals were studied, more females than males were anti-ATLA positive and the antibody-positive rate increased with age. In Japanese monkeys, over 70% were seropositive after the age of 10 years. A family study of Japanese monkeys suggested maternal transmission of ATLV or a closely related agent. These seroepidemiological features are consistent with those of Japanese in the ATL-endemic area. Three of 10 rhesus monkeys were found to be anti-ATLA-positive 13 days after arrival in Japan, suggesting that they had been infected with ATLV in Indonesia where they were captured. Thus, ATLV appears to be prevalent among several macaque species as well as in man, and its pathogenetic role in these non-human primates should be explored in relation to ATL in man.

Intrapleural administration of OK432 in cancer patients: activation of NK cells and reduction of suppressor cells.

Abstract: Twelve patients with carcinomatous pleural effusions were treated with single intrapleural (i.pl.) administration of OK432 on day 0 and the effects of i.pl. OK432 on natural killer (NK) cell activity were followed on day 7. Two patients showed no clinical evidence of therapeutic benefit from i.pl. OK432. In the other 10 patients, pleural effusions and/or tumor cells in the effusions had decreased or disappeared by day 7. NK cell activity was markedly low or absent in pleural effusions of untreated patients due to the presence of adherent effusion cells capable of suppressing the maintenance and interferon-induced augmentation of NK cell activity. I.pl. injection of OK432 resulted in enhancement of NK cell activity and abrogation of NK suppressor cell activity in the effusions. On the other hand, blood NK cell activity was not consistently altered by i.pl. OK432. In vitro treatment of effusion mononuclear cells from untreated patients with OK432, but not with interferon, augmented NK cell activity. In addition, adherent effusion cells of untreated patients lost their NK suppressor function following in vitro OK432 treatment. These results suggest that i.pl. administration of OK432 will result in augmentation of NK cell activity and reduction of NK suppressor cell activity in pleural effusions, which could be responsible for the antitumor activity of i.pl. OK432.

Expression of major histocompatibility antigens and nature of inflammatory cellular infiltrate in ovarian neoplasms.

Abstract: Expression of histocompatibility antigens and the intensity of inflammatory cellular infiltrate were evaluated in frozen tissue sections from 70 human ovarian tumors and six normal ovaries using monoclonal antibodies and an avidin-biotin immunoperoxidase technique. In the normal human ovary, surface epithelial cells, mature granulosa cells and lutein cells reacted with anti-HLA-A, B, C (HLA) and beta2-microglobulin antibodies but not with anti-la (la-like, HLA-DR). Stromal cells and granulosa cells of the primordial follicles did not react with any of the antibodies. Among the neoplasms examined, all benign epithelial tumors, 86% of borderline an 81% of malignant epithelial tumors reacted with anti-HLA and/or beta2-microglobulin antibodies. HLA-negative epithelial tumors were of serous or endometrioid types. Although la was not found in normal ovarian surface epithelium, the antigen could be detected in 44% of benign, and 43% of borderline and malignant epithelial ovarian tumors. Mono-nuclear cellular infiltrate was generally scarce in ovarian tumors and consisted mainly of T cells. Malignant epithelial tumors contained significantly more T cells than did benign tumors. More T cells were observed in HLA-positive ovarian tumors than in HLA-negative neoplasms, but the difference did not achieve statistical significance. No correlation could be found between la expression and the intensity of T-cell infiltrate. Significantly more T8 and Leu-3a-positive cells were found in the tumor stroma than amongst neoplastic cells. HNK-I-positive natural killer cells, OK-MI-positive macrophages and BI-positive B lymphocytes were rarely encountered either in the tumor stroma or between adjacent tumor cells.