Showing papers by "Memorial Sloan Kettering Cancer Center published in 1975"

New Human Tumor Cell Lines

Abstract: There has been a feeling of frustration among many investigators trying to establish cell lines of human tumor cells. Numerous unsuccessful attempts, many of which have remained unpublished, have led to the conclusion that cell line establishment is controlled by the rare occurrence of a tumor with a built-in potential for long-term in vitro growth and by the application of intriguing culture techniques. The results given in the present chapter may, at least in part, contradict these conclusions. With techniques which in no particular way stand out as unusual, and by using medium and serum combinations, and in some cases added hormones, which are generally known to everyone involved in tissue culture, we have, during a rather short period of time, established a high number of new cell lines. These have originated from primary or metastatic human tumors, solid or effusions. In all fairness, many of our attempts to establish continuous lines have failed. However, the total number of attempts over successes is not truly representative of the frequency with which lines might be established. Many variations must be considered in this respect, including the choice of material, collection procedures, lapse of time between the clinical procedure and preparation for tissue culture, technical competence of assistants, and incidental factors known to everyone working in tissue culture. One thing, however, stands out as a conclusion: the careful attention to all the minute details provided by the dedicated tissue culturist seems to be the most important of all factors and cannot well be substituted for by special recipes or fancy equipment.

Possible role for H--Y antigen in the primary determination of sex.

Abstract: IN mammals, the genetic basis of sex determination and differentiation seems to be simple1,2. A gene or set of genes on the Y chromosome causes the indifferent embryonic gonad to develop as a testis3. Thus an ovary develops in the absence of the Y and a testis in its presence. The testis then secretes testosterone which induces male development of the accessory glands and ducts4,5. The masculinising action of testosterone on its target cells is mediated by the product of a gene on the X chromosome. This product activates all the genes required for manifestation of the male phenotype in response to circulating testosterone1. Evidence for this crucial involvement of the X chromosome in male sexual differentiation comes from a mutation of the relevant gene in the mouse (Tfm), resulting in failure to respond to testosterone. A chromosomally XY animal with this mutant gene develops testes, because the Y chromosome is present, but shows no further male differentiation, thus exhibiting the syndrome known as ‘testicular feminisation’6. This insensitivity to androgen is caused by a mutational deficiency of the nuclear–cytosol androgen-receptor protein not only in mice7–9 but also in man10. Thus the part that the X chromosome plays in male sexual differentiation has been clarified at the level of an individual gene situated on this chromosome.

Immunodepression and malignancy.

Abstract: Publisher Summary This chapter reviews some of the experimental and clinical findings linking immunodepression with malignant development. This material has accumulated for the past two decades and sufficient information, whether contradictory or not, is now available to permit an appraisal of the role of immune functions as the regulators of tumor development in animals and man. The idea that immunodepression enhances malignant development has become an integral part of the experimental support of the immune surveillance theory. In its more formal presentations, the theory of immunological surveillance against neoplasia included some basic points, with variations in terminology, but which have appeared with regularity as an integral part of the theory. The degree of immunodepression obtained in many experiments is quite profound and is clearly of a different magnitude than the immune defects occasionally observed in tumor-bearing humans. If immune mechanisms are important in host resistance to antigenic malignancies, any interference with the normal expression of host immunity should facilitate malignant development. Thus, both clinical and experimental evidence indicates that the diseases or procedures that lower the immune functions in mammals are associated with a higher incidence of malignancies.

Role of stem cell migration in initiation of mouse foetal liver haemopoiesis

Abstract: THEORIES of the development of mammalian hepatic haemopoiesis have polarised around two viewpoints. One hypothesis suggests that multipotential haemopoietic stem cells (HSC) migrate from the yolk sac and colonise the developing hepatic primordia1. According to the other view, hepatic haemopoietic tissue arises from the transformation of liver mesenchyme cells and thus has no direct relationship with vitelline haemopoiesis2. To solve this developmental problem, it is necessary to establish techniques, either in vivo or in vitro, to maintain hepatic tissue in a foetal state. In vitro studies have demonstrated that hepatic haemopoietic tissue can develop if tissues are explanted after the 28-somite stage3. Thus, these experiments did not verify the inductive model2 of hepatic haemopoiesis, although the in vitro conditions may have curtailed haemopoietic induction without altering hepatic differentiation. With an exogenous source of yolk-sac-derived HSC attempts have been made to colonise pre-28-somite hepatic explants in vitro, using the appearance of granulopoiesis as an indication of successful recombination. These experiments were not conclusive since granulopoiesis was observed with yolk sac cells in vitro in the absence of hepatic tissue (G.R.J., unpublished).

Effect of tumour necrosis factor on cultured human melanoma cells

Abstract: CARSWELL et al.1 described a tumour necrosis factor (TNF) in the sera of mice sensitised with BCG and injected with endotoxin. After intravenous injection of this serum into mice bearing a 7-d-old transplant of a tumour originally induced by methylcholanthrene, necrosis of the tumour began within 12–24 h. A similar response was found in mice bearing various other tumours. We have now found that both whole TNF serum and TNF partially purified from mouse serum2 inhibit the growth of a human melanoma cell line.

Antibody patterns to herpesviruses in Kaposi's sarcoma. II. Serological association of American Kaposi's sarcoma with cytomegalovirus.

Abstract: The prominent finding of this extended serologic analysis on American and African Kaposi's sarcoma (KS) patients and appropriately matched control groups is the detection of a specific serologic association of cytomegalovirus (CMV) with American KS patients. All American KS sera contained CMV antibodies and their geometric mean titers (GMT) were significantly higher than those in sera of melanoma patients (GMT ratio k = 5.3 to 7.7 by complement fixation [CF], k = 8.9 by indirect hemagglutination [IHA]) or in sera of age- and sex-matched healthy controls (k = 12.6 to 16.0 by CF, k = 12.6 by IHA). The result is strongly reminiscent of the data obtained previously for European KS. Although the GMT to CMV of African KS patients were similar to the GMT of the American KS groups, their significance cannot be demonstrated due to the high background of CMV infections in the control groups. Complex mechanisms are hypothesized, by analogy with the Epstein-Barr virus (EBV) involvement in Burkitt's lymphoma (BL), for a CMV involvement in the development of KS.

Treatment of acute leukemia in adults

Abstract: Improvement in the management of acute leukemia in adults has not progressed nearly so rapidly as has the treatment of childhood leukemia. One important difference is that most adults have myeloblastic or related forms of the disease (AML), whereas the majority of children have lymphoblastic leukemia (ALL). However, even adults with ALL fail to respond as well to a similar regimen as do children with the same type of leukemia. In a recent series of patients with ALL who were treated with the complex multiple drug "L-2" protocol, the incidence of complete remission in adults was 78% vs. 99% in children, and the median duration of remission was only 24 months in the adults, whereas it has not yet been reached in the children and is projected to be over 4 years. In AML and the related nonlymphoblastic forms of acute leukemia, therapy is still unsatisfactory in both adults and children. With the best current drug treatment schedules, the incidence of complete remission is now better than 50%, but it is often difficult to compare the exact remission rates in different series because of differences in reporting results. In adults treated with the multiple drug "L-6" protocol, the incidence of remission in previously untreated patients was 56% and the median duration of remission was 10 months. The median survival of all patients (responders and non-responders) was 1 year whereas that of responders only was 2 years. It is encouraging that a significant proportion of those patients with AML who have complete remissions now remain in remission for extended periods; about 45% of patients responding to the "L-6" protocol remained in remission over 1 year, and 18% have been in continuous remission for 2 to over 4 years. Even after discontinuing treatment, some patients with AML stay in remission for long periods, and it is possible that some of them may have been cured. If this proves to be true, it becomes of great importance to determine what is different about the patients who do exceptionally well as compared to the majority who continue to die within a year. However, no consistent nor distinctive favorable prognostic features have yet been identified.

Evolutionary conservation of H-Y ('male') antigen.

Abstract: THE male specific (H-Y) antigen of mice was discovered with the observation that within certain inbred strains, females reject male skin grafts, whereas skin grafts exchanged between all other sex combinations are accepted1 (reviewed in ref. 2). It is now established that females sensitised with male skin grafts (or immunised with male spleen cells) produce antibody which is cytotoxic for sperm3 and dissociated male epidermal cells4. Using the sperm cytotoxicity test and the mixed haemadsorption-hybrid antibody (MHA˙HA) test, we demonstrated earlier5 that the H-Y antigen of mice is cross reactive or identical with antigen found in male rats, guinea pigs, rabbits and humans. Since then we have extended our survey to classes other than mammals, and we give evidence here for the occurrence of murine H-Y (or a cross reactive) antigen in the white leghorn chicken (Gallus domesticus) and in two amphibian species, the leopard frog (Rana pipiens) and the South African clawed frog (Xenopus laevis).

Proliferative patterns in colonic mucosa in familial polyposis.

Abstract: Microautoradiographic measurements of TdR3H incorporation into epithelial cells of colonic biopsies were compared in normal and eight index cases with familial polyposis, an inherited trait characterized by the development of colonic polyps and carcinomas. Colonic epithelial cells normally synthesized DNA synthesis was proliferated in the deeper two-thirds of the crypts, while DNA synthesis was repressed in the upper one-third. Patients with familial polyposis and patients with isolated single polyps incorporated TdR into epithelial cells lining the surfaces of polyps and occasionally into surface cells of intervening flat mucosa. Symptom-free members of polyposis families also demonstrated TdR3H incorporation into surface epithelial cells in biopsies of morphologically flat mucosa. The findings indicate similar proliferative behavior of epithelial cells in the formation of all adenomas, be they single isolated polyps or multiple familial adenomatous polyps. A sequence of changes occurs involving a loss of the cells' ability to repress DNA synthesis and the appearance of cells with persistent DNA synthesis at the surface and along the upper portion of colonic crypts. These defects are focal and widespread in the colonic mucosa of those with familial polyposis and are expressed at early age.

Indications for pleurectomy in malignant effusion.

Abstract: One hundred six patients with malignant pleural effusion were treated by pleurectomy; 83 of these were available for a 2-year followup. Sixteen of 83 patients were alive 2 years following the pleurectomy, with a survival range of 2-6 years. The most common neoplasms associated with the development of effusion were carcinoma of the lung in 41 cases, carcinoma of the breast in 33 cases, and mesothelioma in 14. Indications for pleurectomy were: A) failure to control the effusion by tube drainage and instillation of chemical or radioactive agents; B) presence of a trapped lung; and C) presence of malignant effusion at the time of thoracotomy for resection of an intrathoracic tumor.

Non-Hodgkin's lymphoma in children.

Abstract: In a study of non-Hodgkin's lymphoma in children, 104 children were treated and followed at Memorial Sloan-Kettering Cancer Center from 1964 throughout June 1974. Forty-three patients, previously treated and untreated, received a nonspecific group of various chemotherapeutic agents and attained an 11% disease-free survival rate. A second group of 18 previously untreated patients, who received a chemotherapeutic regimen consisting of cyclophosphamide alone, achieved a 33% disease-free survival rate. The last group, 43 previously untreated patients (77% of whom had far advanced disease and 86% of whom had diffuse histological types) who received a new and intensive multiple-drug regimen (the LSA2-L2 protocol) consisting of induction, consolidation, and maintenance phases, has maintained an 81% disease-free survival rate after a median observation time of 21+ months. Although nervous system involvement and recurrence or metastases at any time are poor prognostic factors, initial marrow involvement and the amount of bulky disease are no longer considered negative prognosticators when intensive treatment is initiated immediately after diagnosis, is continued for 2--3 years, and includes radiation therapy to sites of bulky disease and CNS prophylaxis. The LS2-L2 treatment is effective in accomplishing the dual aims of not only increasing the numbers of disease-free patients but also prolonging their survival.

Ly antigens as markers for functionally distinct subpopulations of thymus-derived lymphocytes of the mouse.

Abstract: THYMUS-DERIVED lymphocytes (T cells) have several functions1–3, and in some instances it appears that two sub-populations of T cells interact to amplify their response4,5. But recognition of the diverse functions and interactions of T cells has not been matched by the development of techniques for identifying and separating the different T-cell subpopulations involved. The ability to do so would greatly aid study of the many roles of T cells. We report here experiments with mice that show that antisera to different Ly alloantigens can identify functionally-distinct subpopulations of T cells.

Malignant melanoma of the extremities: a clinicopathologic study using levels of invasion (microstage).

Abstract: A clinicopathologic study was done in 151 patients with malignant melanoma of the extremities who were surgically treated in Memorial Hospital and had 5– to 9-year followup. Microstaging was done according to the depth of invasion, as determined by Clark's levels as well as by direct measurement. This was related to treatment and to clinical course. There was a correlation between the depth of invasion by Clark's levels and the incidence of lymph node metastases in patients with Stage I melanoma who had elective node dissection. The incidence of nodal metastases was 4% for Level II, 7% for Level III, 25% for Level IV, and 70% for Level V. There was a correlation between Clark's level of invasion and survival after surgery. The 5-year cure rate was 100% for Level II, 88% for Level III, 60% for Level IV, and 15% for Level V melanoma. The presence of nodal metastases augured a much worse prognosis than Clark's level per se. In patients with Level IV melanoma, the 5-year cure rate was 82% in patients with negative nodes and 27% in those with nodal metastases after elective node dissection. Microstaging primary melanoma according to Clark's levels serves as a useful standard with which to compare surgical results. In this series of extremity melanomas there was no difference between local recurrence and lymphadenectomy for Level II melanoma. For Level III and Level IV melanoma, wide excision and lymphadenectomy gave higher cure rates than wide excision only, both at 5 and 9 years after surgery. The results were significant only for patients with Level III, however. Use of the measured depth of invasion added significant clinicopathologic information. The incidence of nodal metastases at elective node dissection was 5 to 9% for melanoma showing 0.6 to 2.0 mm of invasion, 22% for melanoma measuring 2.1 to 3.0 mm, and 39% for melanoma invading beyond 3.0 mm. The 5-year cure rate was 100% for melanoma measuring less than 1.0 mm, 83% for melanoma invading 1.1 to 2.0 mm, 58% for lesions measuring 2.1 to 3.0 mm, and 55% for melanoma invading over 3.0 mm. The microstage technique combining Clark's levels and the measured depth of invasion has an important use as a prognostic index and as a standard upon which to select treatment for primary melanoma of the extremities.

Multidisciplinary treatment of embryonal rhabdomyosarcoma in children.

Abstract: Twenty-nine children under 15 years of age with embryonal rhabdomyosarcoma were treated according to a multidisciplinary protocol (T-2). The protocol consisted of surgical removal of the tumor if possible, followed by chemotherapy, and also with radiation therapy in patients with gross or microscopic residual disease. Radiation therapy was given in the 4500-7000 rads range. The chemotherapy consisted of cycles of sequential administration of dactinomycin, Adriamycin, vincristine, and cyclophosphamide, with obligatory periods of rest. The drug therapy was continued for 2 years. Following surgery, clinicopathologic staging of the disease revealed 10 patients with no residual disease (I-A), 5 with microscopic residual disease (I-B), 5 with unresectable tumors (II), 6 with unresectable tumors plus regional lymph node involvement (III), and 3 with disseminated tumors (IV). Twenty-four (82%) of the patients (20 Stages I-II, 4 Stage III) are alive with no evidence of disease for 4 plus to 42 plus months. These results are superior to those achieved between 1960-1970 among 108 children treated at Memorial Sloan-Kettering Cancer Center.

Primary carcinoma of the ureter: a prognostic study.

Abstract: Forty-one patients with primary invasive carcinomas of the ureter were seen at Memorial Hospital from 1947 to 1972. Overall survival patterns were similar in 19 patients with and 22 patients without prior or concomitant urothelial cancers elsewhere in the urinary tract, with 5-year survival rates, as estimated by the product-limit method, of 41% for both groups. Prognosis was determined primarily by anatomical stage of ureteral cancer. In 11 Stage A (submucosal) patients, 7 Stage B (muscular), 12 Stage C (periureteric fat), and 9 Stage D (extraureteral), the similarly estimated 5-year survival rates were 91%, 43%, 23%, and nil, respectively. None of Stage A cases had metastases for periods ranging from 5 to 11 years after surgery alone. Seventy-eight percent of patients with more advanced stages died within 3 years of treatment, with metastases mainly in pelvic and para-aortic lymph nodes.

Biochemical evidence linking the GIX thymocyte surface antigen to the gp69/71 envelope glycoprotein of murine leukemia virus.

Abstract: It is known that the thymocyte surface antigen GIX is found in some strains of mice and not others, and that its expression in mice of strain 129, in which most extensive genetic studies have been made, is controlled by two unlinked cellular chromosomal loci. We have now isolated a protein with a mol wt of approximately 70,000 daltons from the surface of thymocytes from 129 mice, which have antigenic and biochemical properties characteristic of the gp69/71 envelope component of murine leukemia virus. Our evidence is compatible with the conclusion that it carries the GIX antigen.

The rationale for multiple drug chemotherapy in the treatment of osteogenic sarcoma.

Abstract: Based on our prior experience in treating children with metastatic osteogenic sarcoma, a multidrug regimen was developed. Nine children with evaluable osteogenic sarcoma were treated with vincristine 1.5 mg/m2 on day 1, highdose methotrexate 200-300 mg/kg i.v. on day 2, with p.o. citrovorum factor "rescue" 9 mg every 6 hours x 12, followed in 2 weeks by cyclophosphamide 40 mg/kg i.v., then 2 weeks later Adriamycin 1.5 mg/kg/day x 2; in 2 weeks cyclophosphamide was repeated. After a 2-week rest, the 56-day cycle was repeated for a total period of 1 year. Oropharyngeal mucositis was the most frequent severe manifestation of gastrointestinal toxicity. Hematologic depression was mild to severe. Nine patients with clinically evaluable osteogenic sarcoma and no previous chemotherapeutic treatment were treated with this regimen. One patient had only a transient shrinkage in tumor mass, and one patient had no progression of multiple pulmonary and bone metastases for 16 months while on therapy. Of the remaining seven patients, all had clinically significant responses with tumor regression demonstrated for from 5 to 20+ months. Four of these patients (three presenting with primary tumor and pulmonary metastases) demonstrated regression of their primary tumor. In an attempt to increase the cure rate in osteogenic sarcoma, chemotherapy that has proven to be effective against metastatic osteogenic sarcoma should now be employed as prophylactic therapy, after amputation, at cancer treatment centers where it can be safely and effectively administered.

Molecular conservation of 74 amino acid sequence of ubiquitin between cattle and man

Abstract: UBIQUITIN (originally ‘ubiquitous immunopoietic polypeptide’ or ‘UBIP’)1 induces lymphocyte differentiation and was first found in bovine thymus during isolation of the thymic polypeptide hormone thymopoietin (originally ‘thymin’)2 which induces T-cell differentiation3 and has secondary effects on neuromuscular transmission2. Ubiquitin, which has a molecular weight of 8,451, is interesting for several reasons. First, it induces differentiation of T cells and, unlike thymopoietin, B cells1. Second, both properties are inhibited by the β-adrenergic blocking agent propranolol, and so the capacity of ubiquitin to induce lymphocyte differentiation seems to depend on reaction with a β-adrenergic receptor. Third, ubiquitin activates adenyl cyclase in lymphocyte precursors and in other tissues, again through a, β-adrenergic receptor (M. Bitensky, and G. G., unpublished). Finally, ubiquitin, as the name implies, is found not only in thymus but in all other animal cells and in yeasts, bacteria and higher plants1. Although its physiological function remains unknown, ubiquitin must be vital to the living cell to have been conserved over such a long evolutionary time span. We have reported the complete primary structure of bovine ubiquitin4—a single polypeptide chain of 74 amino acids—and now report that human ubiquitin has an identical sequence.

Differentiation of T cells in nude mice.

Abstract: Cells bearing the T-cell differentiation alloantigens TL and Thy-1 were enumerated in preparations of spleen and lymph node cells of nu/nu mice. Healthy nu/nu mice had few or no demonstrable TL+ or Thy-1+ cells whereas mice with viral hepatitis, including some born of nu/nu X nu/nu matings, had many. Healthy nu/nu mice were treated daily with the thymic hormone thymopoietin or with ubiquitin, polypeptides that induce the differentiation of TL+Thy-1" cells from TL-Thy-1- precursors in vitro. After 14 days, 20 to 40 percent of their spleen cells were of the TL+Thy-1+ phenotype typical of thymocytes and 10 to 25 percent of their lymph node cells were TL-Thy-1+, typical of later T-cell differentiation. Similar frequencies of such cells were found in untreated nu/nu mice suffering from severe viral hepatitis. These data conform to the current view that prothymocytes are preprogrammed cells whose maturation to thymocytes, normally induced in the thymus by thymopoietin, can be triggered by other agents under abnormal circumstances. Tests of T-cell function were not included in this study.

Lymphocyte differentiation from precursor cells in vitro

Abstract: Much of the recent work in our laboratory has centered about an induction assay developed by Komuro and Boyse in which one of the early maturational steps in the T-cell development, the differentiation from prothymocyte to early thymocyte, can be induced in vitro within two hours. This assay is based on a decade of work concerned with the characterization of cell-surface markers, especially the cytotypic differentiation alloantigens of murine T-cells. As a result of this work on the cell-surface composition of murine lymphocytes, we can summarize the antigenic phenotype of the cortical thymocyte as shown in FIGURE 1. The maturational step in the pathway of T-cell differentiation that can be induced in vitro involves the appearance of this characteristic antigen profile on the surface of the precursor cell; it can be seen that in this differentiation event the products of at least six unlinked genes are expressed. This rapid in vitro T-cell induction assay clearly had potential as a possible bioassay for the identification of the thymic hormone responsible for inducing T-cell differentiation in vivo. Our further studies were designed to answer pertinent questions with respect to this possible application of the assay.

Temperature-sensitive mutants of influenza WSN virus defective in virus-specific RNA synthesis.

Abstract: Influenza WSN virus temperature-sensitive (ts) mutants were examined for defects in viral complementary RNA (cRNA) synthesis. The synthesis of viral cRNA was determined by hybridizing RNA from infected cells to radiolabeled virion RNA of known specific activity. Mutants in complementation groups I and III synthesized little, or no, cRNA at the nonpermissive temperature (39.5 C). When cells infected by these mutants were incubated for 5 h at the permissive temperature (33 C) and were then shifted to 39.5 C, net synthesis of cRNA ceased. This strongly suggests that mutants in these two complementation groups possess a ts defect in the transciptase complex. Mutants in group II and group V synthesize reduced amounts of cRNA at 39.5 C. In contrast to the group I and group III mutants, cRNA synthesis in cells infected by a group II or a group V mutant continues after a shift-up. This indicated that these mutants do not possess a ts transcriptase complex and that these mutants are most probably defective in some step in the amplification of cRNA synthesis. As will be discussed, the most likely defect in these mutants is in the synthesis of virion-type RNA. These results suggest that there are two influenza viral gene functions required for transcription and most likely two additional gene functions required for RNA replication.