skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Thymidine secretion by hybridoma and myeloma cells

Abstract

Secretion of thymidine appeared to be a common property of hybridoma and myeloma cells, but not of other cell types, which were tested. Of three hybridoma cell lines tested, all secreted thymidine in amounts resulting in the accumulation of thymidine to concentrations of 10-20 {mu}M in the culture medium. Also three of five myeloma cell lines that were analyzed secrete thymidine, but none of the other cell types that were studied. Thymidine was purified to homogeneity (4 mg purified from 3 l of culture medium) and identified as such by nuclear magnetic resonance spectroscopy. The cells that secreted thymidine showed high resistance to the growth inhibitory effect of thymidine.

Authors:
 [1];  [2];  [2];  [3]
  1. Department of Molecular Biosciences, University of Oslo, Oslo (Norway). E-mail: bjorn.spilsberg@biokjemi.uio.no
  2. Department of Chemistry, University of Oslo, Oslo (Norway)
  3. Department of Molecular Biosciences, University of Oslo, Oslo (Norway)
Publication Date:
OSTI Identifier:
20798872
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 342; Journal Issue: 1; Other Information: DOI: 10.1016/j.bbrc.2006.01.120; PII: S0006-291X(06)00215-4; Copyright (c) 2006 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ANIMAL GROWTH; CULTURE MEDIA; NUCLEAR MAGNETIC RESONANCE; SECRETION; SPECTROSCOPY; THYMIDINE

Citation Formats

Spilsberg, Bjorn, Rise, Frode, Petersen, Dirk, and Nissen-Meyer, Jon. Thymidine secretion by hybridoma and myeloma cells. United States: N. p., 2006. Web. doi:10.1016/j.bbrc.2006.01.120.
Spilsberg, Bjorn, Rise, Frode, Petersen, Dirk, & Nissen-Meyer, Jon. Thymidine secretion by hybridoma and myeloma cells. United States. doi:10.1016/j.bbrc.2006.01.120.
Spilsberg, Bjorn, Rise, Frode, Petersen, Dirk, and Nissen-Meyer, Jon. Fri . "Thymidine secretion by hybridoma and myeloma cells". United States. doi:10.1016/j.bbrc.2006.01.120.
@article{osti_20798872,
title = {Thymidine secretion by hybridoma and myeloma cells},
author = {Spilsberg, Bjorn and Rise, Frode and Petersen, Dirk and Nissen-Meyer, Jon},
abstractNote = {Secretion of thymidine appeared to be a common property of hybridoma and myeloma cells, but not of other cell types, which were tested. Of three hybridoma cell lines tested, all secreted thymidine in amounts resulting in the accumulation of thymidine to concentrations of 10-20 {mu}M in the culture medium. Also three of five myeloma cell lines that were analyzed secrete thymidine, but none of the other cell types that were studied. Thymidine was purified to homogeneity (4 mg purified from 3 l of culture medium) and identified as such by nuclear magnetic resonance spectroscopy. The cells that secreted thymidine showed high resistance to the growth inhibitory effect of thymidine.},
doi = {10.1016/j.bbrc.2006.01.120},
journal = {Biochemical and Biophysical Research Communications},
number = 1,
volume = 342,
place = {United States},
year = {Fri Mar 31 00:00:00 EST 2006},
month = {Fri Mar 31 00:00:00 EST 2006}
}
  • Highlights: • Levels of EEN expression paralleled with the rate of cell proliferation. • EEN was involved in the proliferation and survival of multiple myeloma (MM) cells. • EEN regulated the activity of IGF-1-Akt/mTOR pathway. • EEN regulated proliferation and survival of MM cells by enhancing IGF-1 secretion. - Abstract: The molecular mechanisms of multiple myeloma are not well defined. EEN is an endocytosis-regulating molecule. Here we report that EEN regulates the proliferation and survival of multiple myeloma cells, by regulating IGF-1 secretion. In the present study, we observed that EEN expression paralleled with cell proliferation, EEN accelerated cell proliferation,more » facilitated cell cycle transition from G1 to S phase by regulating cyclin-dependent kinases (CDKs) pathway, and delayed cell apoptosis via Bcl2/Bax-mitochondrial pathway. Mechanistically, we found that EEN was indispensable for insulin-like growth factor-1 (IGF-1) secretion and the activation of protein kinase B-mammalian target of rapamycin (Akt-mTOR) pathway. Exogenous IGF-1 overcame the phenotype of EEN depletion, while IGF-1 neutralization overcame that of EEN over-expression. Collectively, these data suggest that EEN may play a pivotal role in excessive cell proliferation and insufficient cell apoptosis of bone marrow plasma cells in multiple myeloma. Therefore, EEN may represent a potential diagnostic marker or therapeutic target for multiple myeloma.« less
  • Human T lymphocytes are capable of secreting many different lymphokines which regulate Ig production. The authors produced a human T-T fusion between T cells from a normal donor and the T lymphoma line CEM-6. From this fusion 3 of 180 clones produced a factor which suppressed polyclonal Ig production by PWM-stimulated mononuclear cells. All 3 clones expressed both T4 and T8 surface markers; CEM-6 is T4/sup +//T8/sup +/. Suppression of Ig production did not involve cytotoxicity and was most effective when the hybridoma supernatant was added during the first 3 d of culture. Production of the suppressive factor could bemore » inhibited by corticosteroids but not indomethacin. Analysis of the hybridoma supernatant by reverse phase chromatography revealed a peak of suppressive activity in the fraction where purified LTC/sub 4/ could be eluted. Crude hybridoma supernatant contained 10-50 ng/ml of LTC/sub 4/, but no LTB/sub 4/ by radioimmunoassay (RIA). In addition, LTC/sub 4/ produced by the hybridomas co-eluted with purified LTC/sub 4/ as assessed by RIA. No LTC/sub 4/ was found in nonsuppressive hybridoma supernatants. Furthermore, antibody to LTC/sub 4/ could absorb suppressive activity from hybridoma supernatants. Thus, human T lymphocytes produce LTC/sub 4/ which suppresses polyclonal Ig production by human mononuclear cells.« less
  • Although effects of nickel(II) on the immune system have long been recognized, little is known about the effects of nickel(II) on the induction of apoptosis and related signaling events in T cells. In the present study, we investigated the roles and signaling pathways of nickel(II) in the induction of apoptosis in a human T cell line jurkat. The results showed that the cytotoxic effects of Ni involved significant morphological changes and chromosomal condensation (Hoechst 33258 staining). Analyses of hypodiploid cells and FITC-Annexin V and PI double staining showed significant increase of apoptosis in jurkat cells 6, 12 and 24 hmore » after nickel(II) treatment. Flow cytometry analysis also revealed that the loss of mitochondrial membrane potential (MMP) occurred concomitantly with the onset of NiCl{sub 2}-induced apoptosis. Induction of apoptotic cell death by nickel was mediated by reduction of bcl-2 expression. Furthermore, nickel stimulated the generation of nitric oxide (NO). These results suggest that nickel(II) chloride induces jurkat cells apoptosis via nitric oxide generation, mitochondrial depolarization and bcl-2 suppression.« less
  • In vivo S-phase cell labeling with iododeoxyuridine (IdUrd) was performed in six multiple myeloma (MM) patients. Myeloma cells from four patients were hyperploid. In three out of four patients, DNA/IdUrd flow cytometry revealed that most of the labeled cells, which had divided during the period, elapsed between flash labeling and sampling, had returned to the diploid G0/G1 compartment and not to the hyperdiploid peak. To eliminate contaminating cells belonging to the normal hematopoiesis, plasmocytic and lymphocytic cells were fractionated and analyzed separately. Cell enrichment was performed with use of murine monoclonal antibodies (MoAbs) against plasmocytic and lymphocytic cell markers andmore » subsequent magnetic activated cell sorting with immunobeads, i.e., polystyrene magnetic particles coated with sheep anti-mouse IgG. The IdUrd-labeled cells were predominantly lymphocytic cells, returning after mitosis to the diploid G0/G1 peak. Although this pattern of S-phase cells in hyperdiploid MM, belonging to the diploid cell compartment, was observed in three out of four hyperploid cases and although the number of observations is small, S-phase cells may demonstrate an aspect of tumor cell kinetics in hyperploid MM, which has been debated for many years and which indicates the existence of a non-plasmocytic stem cell compartment that feeds the plasmocytoma. The behavior of the labeled cells as observed in a few cases of MM provides another, hitherto undescribed, argument that, at least in some MM patients, a part of the proliferating tumor cells may be diploid lymphocytic (precursor) cells. These findings should be considered when targeting and monitoring treatment of MM and also in purging procedures of bone marrow in patients to be treated by ablative cytotoxic therapy and autologous bone marrow transplantation. 57 refs., 3 figs., 1 tab.« less