Showing papers by "Fred Hutchinson Cancer Research Center published in 1980"
TL;DR: Two new monoclonal antibodies that react with surface antigens of human lymphocytes are described that were identified on the framework region of a T-cell antigen and designated HuLyt-1 and 9.3.
Abstract: We describe here two new monoclonal antibodies that react with surface antigens of human lymphocytes. Antibody 7.2 identified a determinant on the framework region of the human Ia antigen. It was cytotoxic for all cultured B-cell lines, normal B cells, and monocytes. The antibody was not cytotoxic for normal T cells or for established T leukemic cell lines. In immune precipitation assays, the 7.2 antibody reacted with a bimolecular complex of two chains that resolved in polyacrylamide gels as polypeptides with molecular weights of 29000 and 34000 daltons. These precipitation results were analogous to those achieved with a rabbit antiserum prepared against human Ia antigens. Antibody 9.3 identified a determinant on the framework region of a T-cell antigen. It was cytotoxic for 50–80% of peripheral T cells and for 20–50% of thymocytes. It was not cytotoxic for cultured B-cell lines, normal B cells, or monocytes. In immune precipitation assays, the 9.3 antibody reacted with a single polypeptide with a molecular weight of 44000 daltons. Due to the expression of this antigen on a limited subpopulation of human T cells, we have designated the antigen HuLyt-1.
360 citations
TL;DR: Analysis of coded liver histology from 204 consecutive autopsied recipients transplanted for leukemia showed a significantly higher prevalence of venocclusive disease associated with transplantation for leukemia, pretransplant conditioning with more rigorous chemoradiotherapy regimens, and statistical analyses showed.
265 citations
TL;DR: Immunoprecipitation and sodium dodecyl sulfate gel electrophoresis were used to detect the target antigen in 125I-labeled cell membranes of both cultivated cells and tumor biopsy samples, and a protein with a molecular weight of 97,000 was identified.
Abstract: BALB/c mice were immunized with a human melanoma cell line, SK-MEL 28, and their spleen cells were fused with mouse NS-1 myeloma cells. Hybrid cells were tested in an indirect 125I-labeled protein A assay for production of antibodies that bound to surface antigens of SK-MEL 28 melanoma cells but not to autologous skin fibroblasts. One hybridoma, designated 4.1, had the required specificity. It was cloned and grown in mice as an ascites tumor. The monoclonal IgG1 antibody produced by the hybridoma was purified from the ascites fluid and labeled with 125I. The labeled antibody bound, at significant levels, to approximately 90% of the melanomas tested and to approximately 55% of other tumor cells, but not to three B-lymphoblastoid cell lines or to cultivated fibroblasts from 15 donors. Immunoprecipitation and sodium dodecyl sulfate gel electrophoresis were used to detect the target antigen in 125I-labeled cell membranes of both cultivated cells and tumor biopsy samples. A protein with a molecular weight of 97,000 was identified. This protein, designated p97, was present in both cultured cells and biopsy material from melanomas and certain other tumors, but it was not detected in eight different samples of normal adult epithelial or mesenchymal tissues obtained from five donors.
251 citations
TL;DR: Direct evidence is reported here direct evidence that immunoglobulin-synthesizing B lymphocytes can arise from the CML stem cell clone.
Abstract: Studies with glucose-6-phosphate dehydrogenase (G6PD) iso-enzymes have demonstrated that chronic myelogenous leukaemia (CML) is a clonal disorder of pluripotent haematopoietic stem cells which are capable of differentiation to myeloid cells, monocytes, erthrocytes and platelets1. It has been observed recently in G6PD heterozygous patients with chronic phase CML that the non-E-rosetting lymphocytes were restricted to a single enzyme type, indicating that some lymphoid cells must also arise from the leukaemic clone2. Surface or cytoplas-mic immunoglobulin could be detected in up to 46% of the cells of these isolated non-T-lymphocyte populations, which suggested that cells from the CML clone were capable of differentiating into B lymphocytes. To investigate this further, we established Epstein–Barr virus (EBV)-transformed B-lymphoblastoid cell lines derived from patients with CML and studied chromosomes and G6PD to determine whether progenitor B lymphocytes for any of the cell lines had originated from the CML clone. We report here direct evidence that immunoglobulin-synthesizing B lymphocytes can arise from the CML stem cell clone.
246 citations
TL;DR: Recovery of responsiveness to HSV antigen after marrow transplant is primarily related to recurrent virus infection and not to the pretransplant immune status of the donor.
Abstract: The relationship between herpes simplex virus (HSV) infection and specific cell-mediated immunity was investigated in 141 patients before and for the first four months after marrow transplant. Sixty-two (82%) of 76 seropositive patients but only one of 65 seronegative patients developed HSV infection. Lymphocyte responses to HSV antigen were suppressed immediately after transplant and subsequently became reactive in those patients with HSV infection. The presence or absence of antibody to HSV in the donor before transplant did not influence the response. Seventy long-term survivors of marrow transplant were also studied. Among 60 patients who had pretransplant serum available for study, 26 (68%) of 38 who had been seropositive before transplant had positive responses compared with none of 22 who had been seronegative. Recovery of responsiveness to HSV antigen after marrow transplant is primarily related to recurrent virus infection and not to the pretransplant immune status of the donor.
202 citations
TL;DR: Treatment patterns revealed that surgery remains the most common treatment modality, and review of patterns of survival suggested that children in the other nonwhites category with unilateral disease had poorest survival rates.
Abstract: • Data from the population-based Surveillance, Epidemiology, and End Results Program of the National Cancer Institute were used to calculate the incidence of retinoblastoma for the years 1974 through 1976. Each year 3.58 cases occurred for each million children under the age of 15 years. Incidence was markedly age related, with over 90% of the cases being diagnosed before the age of 5 years. Although no difference in incidence was found for whites and blacks, other nonwhites had rates greater than four times those of whites. Twenty percent of patients had bilateral disease. Treatment patterns revealed that surgery remains the most common treatment modality. Review of patterns of survival suggested that children in the other nonwhite category with unilateral disease had poorest survival rates.
189 citations
TL;DR: Monoclonal antibodies against a thymus cell differentiation antigen (Thy-1.1) were effective in the therapy of a transplanted mouse leukemia and resulted in the cure of leukemia in a significant proportion of the treated animals.
Abstract: Monoclonal antibodies against a thymus cell differentiation antigen (Thy-1.1) were effective in the therapy of a transplanted mouse leukemia. Passive immunization resulted in high titers of cytotoxic antibody in the serum of treated mice and the suppression of metastatic tumor cells. The tumor-suppressive effects of the monoclonal antibodies were amplified by the administration of exogenous complement. This combined antibody and complement therapy resulted in the cure of leukemia in a significant proportion of the treated animals.
188 citations
TL;DR: Spleen cells from a mouse immunized with human melanoma cells were fused with mouse myeloma cells, and somatic cell hybrids were grown in selective medium, and seven of the eight melanoma proteins identified were present at the cell surface.
163 citations
TL;DR: Immune precipitates formed with 10.2 antibody and detergent lysates of radiolabeled T-cells contained three polypeptides with molecular weights of 65 000, 55 000, and 50000 daltons, but it has not been determined whether these proteins represent a multimeric antigen complex.
Abstract: Monoclonal antibody 10.2 reacts with a monomorphic antigen expressed on the surface of virtually all thymocytes, as well as thymus-dependent lymphocytes in the peripheral blood and bone marrow. In contrast, antibody 10.2 did not react with normal peripheral blood B cells, monocytes, or the non-T-cell fraction of bone marrow. This complement fixing IgG2a antibody also reacted with extablished leukemic T-cell lines, but not with cell lines of either normal or malignant B-cell origin. Similarly, when tested against acute leukemia blasts, the 10.2 antibody reacted with those from patients with T-cell acute leukemia, but not with those from patients with acute null cell or non-lymphocytic leukemia. An unexpected exception to this pattern was the reaction of 10.2 antibody with leukemic cells from patients with B-cell type chronic lymphocytic leukemia. Immune precipitates formed with 10.2 antibody and detergent lysates of radiolabeled T-cells contained three polypeptides with molecular weights of 65 000, 55 000, and 50000 daltons. It has not been determined whether all three of these polypeptides contain the 10.2 antigenic determinant, or whether these proteins represent a multimeric antigen complex.
152 citations
TL;DR: It was concluded that recovery of the response to CMV antigen is related primarily to active virus infection and not to patient or donor pretransplant immunity.
Abstract: Patients were studied prospectively after marrow transplant to correlate cytomegalovirus (CMV) infection with the in vitro lymphocyte transformation response to CMV antigen. Ninety-two (58%) of 158 patients developed CMV infection. The lymphocyte response to CMV antigen of patients who were seropositive before transplant was significantly suppressed immediately after transplant. Isolation of CMV was associated with further suppression of responses; seroconversion to CMV was associated with a significant increase. The lymphocyte response of 73 long-term survivors was similar to that of normal persons. The presence of antibody to CMV in the donor before transplant had little effect on the lymphocyte response of patients after transplant even though the patients' lymphocyte s were of donor origin. As in previously reported studies of immunity to other herpesviruses after marrow transplant, it was concluded that recovery of the response to CMV antigen is related primarily to active virus infection and not to patient or donor pretransplant immunity.
135 citations
TL;DR: Under simplifying assumptions the quantitative effect of migration on risk estimates is shown as a function of cancer site, latent period, and the type of geographic units for which rates are calculated--states, counties, or places.
Abstract: Cancer rates are often compared between counties or other geographic units as a method of testing for risk from environmental exposures. Migration between geographic areas greatly reduces the sensitivity of this method. Under simplifying assumptions the quantitative effect of migration on risk estimates is shown using migration and cancer incidence data for the United States. For example, 40--50% of the relative excess risk, defined as the relative risk minus one, is not reflected in the estimated risk for most cancers, when rates are compared between exposed and unexposed counties and migration has taken place during a 30-year latent period. More extreme losses of sensitivity also occur. Under the simplifying assumptions, the quantitative effect of migration on risk estimates is shown as a function of cancer site, latent period, and the type of geographic units for which rates are calculated--states, counties, or places. Also discussed are some implications of these findings for geographically-based studies and additional data needs for assessing the effect of migration.
TL;DR: Findings point to the potential use of human-mouse hybrid cells as a source of human monoclonal antibodies for therapeutic and diagnostic purposes.
Abstract: Hybrid cells formed between human lymphocytes and mouse myeloma cells produce human immunoglobulin in culture. Stable antibody-producing cell lines can be isolated after multiple cycles of low-density passage, cloning, and continued selection for immunoglobulin production. The origin and characteristics of a hybrid of human and mouse cells is described. This hybrid produces high concentrations (8.3 micrograms per milliliter) of human immunoglobulin M reactive with the terminal disaccharide of the Forssman glycolipid. These findings point to the potential use of human-mouse hybrid cells as a source of human monoclonal antibodies for therapeutic and diagnostic purposes.
TL;DR: The present experiments indicate that LLV-induced tumours contain transforming genes that can be detected by transfection of NIH 3T3 mouse cells, suggesting that oncogenesis by LLVs may result from indirect activation of cellular transforming genes.
Abstract: Oncogenic avian retroviruses can be classified into three groups: sarcoma viruses, acute leukaemia viruses and lymphoid leukosis viruses (LLVs)1. Sarcoma and acute leukaemia viruses transform fibroblasts and/or haematopoietic cells in culture and induce tumours with short latent periods in infected birds. In contrast, LLVs do not transform cells in vitro and require long latent periods before formation of neoplasms in vivo. The most frequent neoplasm induced by LLVs is malignant lymphoma of the bursa of Fabricius, but LLVs also induce other neoplasms, including sarcomas, nephroblastomas and erythroblastosis2. The genomes of both sarcoma and acute leukaemia viruses contain specific genes responsible for viral oncogenicity1,3–5, whereas the genome of LLVs apparently includes only genes required for virus replication. The genetic basis for the low oncogenic potential of LLVs is therefore obscure. The present experiments indicate that LLV-induced tumours contain transforming genes that can be detected by transfection of NIH 3T3 mouse cells. These transforming genes are not linked to LLV DNA sequences, suggesting that oncogenesis by LLVs may result from indirect activation of cellular transforming genes.
TL;DR: 3H‐HSV‐2 DNA has been hybridized in situ to frozen sectioned tissue from human cervical biopsies and RNA complementary to the virus‐specific probe was detected in cells undergoing pre‐malignant changes, but not in the cells of the fully developed squamous‐cell cancer.
Abstract: 3H-HSV-2 DNA has been hybridized in situ to frozen sectioned tissue from human cervical biopsies. RNA complementary to the virus-specific probe was detected in cells undergoing pre-malignant changes, but not in the cells of the fully developed squamous-cell cancer.
TL;DR: Eight recipients of marrow transplants with cytomegalovirus pneumonia diagnosed by open lung biopsy were treated with doses of human leukocyte interferon to evaluate its toxicity after marrow transplant and its effectiveness against CMV infection, and there was less hematologic toxicity than was anticipated.
Abstract: Eight recipients of marrow transplants with cytomegalovirus (CMV) pneumonia diagnosed by open lung biopsy were treated with doses of human leukocyte interferon of 2 X 10(4)--6.4 X 10(5) units/kg per day to evaluate its toxicity after marrow transplant and its effectiveness against CMV infection. All eight patients died from pneumonia, and virus was still present in lung tissue from seven patients cultured after death. Interferon doses of less than or equal to 1.6 X 10(5) units/kg per day did not affect the circulating granulocyte count. Patients treated with higher doses had a decrease in circulating granulocyte count, but study of granulocyte-macrophage colony-forming cells in culture showed no evidence of toxicity to granulocyte progenitor cells. The effect on in vitro lymphocyte function was variable. Antibody production was not impaired. Interferon was not effective against established CMV infection. However, there was less hematologic toxicity than was anticipated, and the prophylactic use of interferon after marrow transplant is feasible.
TL;DR: Methodologic comparisons between the major studies and suggestions for future research were made to clarify the inconsistent evidence of the benefits from breast self-exam on extent of disease.
TL;DR: The lymphocyte response to VzV antigen parelleled, and thus may predict, periods of increased susceptibility to VZV infection.
Abstract: The cellular immune response of normal persons and marrow transplant recipients to varicella-zoster virus (VZV) antigen was measured with use of the lymphocyte transformation response. Ninety-nine of 100 normal persons with previous VZV infection had a stimulation index of greater than or equal to 4.8, while 10 susceptible persons had responses of less than or equal to 3.0. Before transplant, responses were lower than normal in patients with leukemia in relapse (P = 0.001), but not in patients with leukemia in remission or with aplastic anemia. Throughout the first 100 days after transplant, lymphocyte response was depressed (P less than 0.0005), especially among recipients of antithymocyte globulin during days 41-80 (P less than 0.05). Patients with aplastic anemia had higher responses than those with leukemia during days 20-60 (P less than 0.02). By one year, most responses were normal. Long-term survivors who had recurrent VZV infection had positive responses more often than those without recurrent infection (P = 0.01). The lymphocyte response to VZV antigen parelleled, and thus may predict, periods of increased susceptibility to VZV infection.
TL;DR: Results of these competition binding assays demonstrate the utility of this type of serological analysis for the study of the tertiary structure of individual viral proteins.
TL;DR: Better survival rates have been achieved, and therapy has been refined so that treatment can be reduced in early stage disease without jeopardizing tumor control, which results in fewer short‐ and long‐term complications.
Abstract: Major advances have been made against Wilms' tumor as a result of treatment methods developed by single institutions that then have been confirmed and extended by national cooperating groups. Better survival rates have been achieved, and therapy has been refined so that treatment can be reduced in early stage disease without jeopardizing tumor control. This results in fewer short- and long-term complications, an especially important consideration in children. Their organs and tissues are vulnerable to anti-mitotic treatments such as chemo- and radiotherapy, that can produce disabling if not lethal dysfunctions. This progress has been the result of the cooperative efforts by multiple specialists, and provides evidence of the value of such integrated studies. They have changed the outlook from a 90% death rate in the early years of this century to the 90% survival rate now possible with modern management.
Journal Article•
Journal Article•
TL;DR: A significant discordance was showed between NK, ADCC, and LDCC activities in these immunologically perturbed individuals, indicating that either different cell populations or different cellular mechanisms are involved in these cytotoxic functions.
Abstract: Cytotoxic effector cells mediating natural killing (NK), antibody-dependent cellular cytotoxicity (ADCC), and lectin-dependent cellular cytotoxicity (LDCC) were studied in patients with leukemia or aplastic anemia before and/or after marrow transplantation. Before transplantation, about one-third to one-half of the patients were deficient in cytotoxic activity. In patients with leukemia, this was most likely due to large numbers of circulating blast cells diluting or replacing the effector cells. In patients with aplastic anemia there was an apparent absence of the effector cells in a proportion of the patients. After marrow transplantation, cytotoxic activity in all three systems returned to normal rapidly, by 30 days, and remained so through 100 days. However, about 20% of patients studied beyond 1 year were deficient in these functions. There were no significant associations between cytotoxic activity and important clinical parameters including infections, graft-vs-host disease, and recurrence of leukemia. Our findings do not support an immunosurveillance role for NK against leukemia after marrow transplantation. Furthermore, they point out the need for new in vitro approaches for meaningful monitoring of marrow transplant patients. Finally, our results showed a significant discordance between NK, ADCC, and LDCC activities in these immunologically perturbed individuals, indicating that either different cell populations or different cellular mechanisms are involved in these cytotoxic functions.
TL;DR: It is demonstrated that the globin gene in chicken embryo fibroblasts is activated by infection with two other oncogenic retroviruses, avian erythroblastosis virus and strain MC-29 of avian myeloblastosis virus, which contain transforming genes unrelated in nucleotide sequence content to each other or to the Rous sarcoma virus src gene.
Abstract: We demonstrated previously that chicken embryo fibroblasts accumulate approximately 100 copies of embryonic globin RNA after transformation by Rous sarcoma virus. Here we demonstrate that the globin gene in chicken embryo fibroblasts is activated by infection with two other oncogenic retroviruses, avian erythroblastosis virus and strain MC-29 of avian myeloblastosis virus, which contain transforming genes unrelated in nucleotide sequence content to each other or to the Rous sarcoma virus src gene. In addition, we have measured the genetic complexity of transformation by using established techniques for determining the number of different RNA sequences in specific populations of cells. Our results indicate that transformation of chicken embryo fibroblasts by Rous sarcoma virus results in the accumulation of RNA from approximately 1000 average-sized new transcription units.
TL;DR: A processing scheme for the generation of reverse transcriptase subunits is presented and it is suggested that the alpha subunit is generated by a COOH-terminal cleavage of the beta subunit.
Abstract: We have studied the biosynthesis of avian retrovirus proteins related to reverse transcriptase in permissive avian embryonic cells. Analysis of immune precipitates from avian sarcoma virus (ASV)-infected cells demonstrated the presence of the 180,000-dalton gag-pol "read-through" protein (Pr180gag-pol) and a 130,000-dalton polypeptide (Pr130gag-pol). Pr130gag-pol was found, in serological and peptide mapping studies, to consist primarily of sequences related to reverse transcriptase and the gag-encoded protein p15. Pr180gag-pol was found to be phosphorylated, whereas Pr130gag-pol was not. In addition, only Pr180gag-pol but not Pr130gag-pol was susceptible to cleavage with the virion protease p15. Although the structure of Pr130gag-pol would suggest that it is generated by removal of a portion of the gag region from Pr180gag-pol, an analysis of labeling kinetics has failed to demonstrate unequivocally whether Pr130gag-pol is a cleavage product of Pr180gag-pol or a primary translation product. We were repeatedly unable to detect either Pr180gag-pol or Pr130gag-pol in virus particles released from the cell, whereas both beta and alpha subunits were readily observed. Several presumed intermediates between Pr130gag-pol and the beta subunit of reverse transcriptase were also observed in virions. These studies indicate cleavage of polyemrase precursors at the time of virus budding. On the basis of these data, we present a processing scheme for the generation of reverse transcriptase subunits. We have also examined reverse transcriptase biosynthesis in cells producing two mutants that fail to package the enzyme. Previous work showed that integrated proviruses of both mutants are missing DNA sequences in pol: one mutant, PH9 (Mason et al., J. Virol. 30:132-140, 1979), contains a deletion near the 3' end of pol, whereas the other, SE52d (linial et al., Virology 87:130-141, 1978), may have inserted a host cell sequence near the 5' end of pol. Neither mutant synthesized Pr180gag-pol or Pr130gag-pol, but instead produced novel proteins comprised of sequences shared with gag proteins plus a region antigenically related to reverse transcriptase. Both proteins were defective as precursors to reverse transcriptase. Whereas Pr180gag-pol and Pr130gag-pol were precipitated by an antiserum raised against p32 (a virion protein derived from the portion of the beta subunit removed during processing of beta to alpha [Schiff and Grandgenett, J. Virol. 28:279-291, 1978]), the novel protein synthesized by PH9 ws not precipitated. This suggets that the alpha subunit is generated by a COOH-terminal cleavage of the beta subunit.
Journal Article•
TL;DR: It is demonstrated here that K562 cells express the fetal type (i) antigen on distinctly different glycoproteins from those of erythrocytes, by the use of cell-surface labelling followed by endo-β-galactosidase digestion or followed by immunoprecipitation with specific antibodies.
Abstract: During the ontogenic change from fetal to adult human erythrocytes, as well as fetal haemoglobin being replaced by adult haemoglobin, the cell-surface antigen i is converted to I (ref. 1). Recently it has been shown that this antigenic change is the conversion of the linear repeating Galβ1→4GlcNacβ1→3 Gal structure to branched Galβ1→4 GlcNacβ1→3(Galβ1→4GlcNacβ1→6)Gal structure2–4. We have shown that cell-surface labelling followed by endo-β-galactosidase digestion can distinguish these two forms on the cell surface, and that band 3 and band 4.5 are the major carriers for these antigens on mature erythrocytes5. Human leukaemic cell line K562, originally isolated from a patient at blast crisis of chronic myelocytic leukaemia6, has recently been shown to synthesize glycophorin A7, and to be capable of synthesizing haemoglobin upon induction8,9. I demonstrate here that K562 cells express the fetal type (i) antigen on distinctly different glycoproteins from those of erythrocytes, by the use of cell-surface labelling followed by endo-β-galactosidase digestion or followed by immunoprecipitation with specific antibodies.
TL;DR: Two amphipathic protein fractions soluble in organic solvents as well as in water have been isolated from the ganglioside fraction of bovine erythrocyte membranes by successive chromatography in chloroform-methanol mixture.
01 Jan 1980
TL;DR: Limitations in the effectiveness of serum treatment may have resulted from insufficient quantities of high-titered antibody of appropriate class, avidity, and specificity.
Abstract: Antibody treatment of neoplastic disease has long been of interest. Under certain limited conditions, significant inhibition of tumor growth has been achieved (reviewed in Wright and Bernstein, 1980). The therapeutic effects obtained with antisera have suggested a potential role for antibody therapy, but the success of this approach has so far not been impressive. Limitations in the effectiveness of serum treatment may have resulted from insufficient quantities of high-titered antibody of appropriate class, avidity, and specificity.
TL;DR: It is reported that the stage-specific embryonic antigen (SSEA-1), defined by a monoclonal antibody reacting with early mouse embryos and murine embryonal carcinoma cells, is a complex glycolipid with the branched lacto-N-glycosyl series.
TL;DR: Oligosaccharides released by reductive alkaline degradation of blood group Ii active sheep gastric mucins have been shown to contain two types of branched core structure: the novel structure was the core of one of the main oligosACcharide fractions and the other was identical to that reported previously in various human and animal mucins.