PubMed Health⌕ Search

Biomedical subjects

J F Eliason

Publications and source records attributed to J F Eliason.

At least 19 recordsLinked to original sources
PubMed1992

Responsiveness of three newly established human colorectal cancer cell lines to transforming growth factors beta 1 and beta 2.

We have established 3 new human colorectal cancer cell lines (LS411N, LS513, and LS1034) from clinical biopsy samples. These lines are tumorigenic and grow s.c. as adenocarcinomas in nude mouse xenografts. Specific marker chromosomes are observed in each line. Carcinoembryonic antigen is expressed at the surface of all 3 lines, but with marked quantitative differences. Indeed, less than 10% of the cells from the HT-29 line used as a reference express carcinoembryonic antigen while more than 90% of the LS1034 cells do so. LS513 and LS1034 consistently express HLA class I antigens and intercellular adhesion molecule 1 which are not detected at the surface of the LS411N cells. No expression of HLA class II antigens DR, DQ, and DP has been measured on any of the lines. All three lines grow well in 5% fetal calf serum medium without addition of exogenous growth factors. The LS1034 line has been adapted to growth in serum-free conditions and exhibits increased clonogenicity when cells are seeded in serum-free methylcellulose medium, as compared with medium containing 5% fetal calf serum. The LS513 and LS1034 lines have proved to be of particular interest since they respond to the growth-inhibitory action of TGF-beta 1 and TGF-beta 2 in both liquid and semisolid medium. Both factors were, at pM concentrations, equipotent inhibitors of LS1034 cell proliferation. In contrast, higher concentrations of TGF-beta 1 are inhibitory for proliferation of LS513 cells, whereas TGF-beta 2 has no effect on the growth of these cells in liquid assay. On this basis, using appropriate anti-TGF-beta 1 and anti-TGF-beta 1 IgY, we developed a bioassay for TGF-beta 1 and TGF-beta 2. Two of the three lines have indeed been shown to produce latent-TGF-beta 1 activity.

Adult↗
PubMed1992

Interactions of interferon-alpha 2a with 5'-deoxy-5-fluorouridine in colorectal cancer cells in vitro.

The biological activity of 5'-deoxy-5-fluorouridine (5'-dFUrd) depends upon intracellular enzymatic cleavage by pyrimidine phosphorylase to form 5-fluorouracil (5-FU). Interferon-alpha 2a (IFN-alpha) effect was analysed alone and combined with 5-FU or 5'-dFUrd, on proliferation inhibition of eight human colorectal cancer cell lines. The toxicity of 5-FU was enhanced by IFN-alpha in only one line (SW-480). In contrast, interactive enhancement of IFN-alpha was observed with 5'-dFUrd in five lines (WiDr, HT-29, 513, SW-480 and Co-115). In each of the lines showing potentiation by IFN/5'dFUrd but not by IFN/5-FU, cytoplasmic pyrimidine phosphorylase activity was increased after 5 days' incubation with IFN-alpha in a dose-dependent manner. Two lines (LISP-1 and SW-620) showed no potentiation of either 5-FU or 5'-dFUrd toxicity by IFN-alpha, and no change in pyrimidine phosphorylase activity. Potentiation of 5'-dFUrd effect by IFN-alpha may thus be explained by an enhancement of its conversion to 5-FU through stimulation of pyrimidine phosphorylase activity.

Antineoplastic Agents↗
PubMed1990

Binding, internalization and degradation of radio-iodinated interleukin 3 and granulocyte-macrophage colony stimulating factor by various hemopoietic cells.

The proliferation and differentiation of hemopoietic committed progenitor cells depend on colony stimulating factors (CSF). However, isolated mouse granulocyte-macrophage progenitor cells can still undergo limited proliferation in serum-free cultures after CSF deprivation. To test whether this is due to an accumulated pool of internalized factor, we examined the binding, internalization and degradation of radiolabelled interleukin 3 (IL-3) and granulocyte-macrophage colony stimulating factor (GM-CSF) in various hemopoietic cells. We found 20,000 high affinity IL-3 receptors on cells of two IL-3-dependent hemopoietic cell lines, FDC-P1 and FDC-P2 (Kd = 85 and 129 pM). FDC-P1 cells, which also respond to GM-CSF, possess 600 high-affinity GM-CSF receptors (Kd = 64 pM). Cells of both lines internalize IL-3, but only FDC-P1 cells release degraded IL-3 at a rapid rate. Both cell lines have similar dose-response curves for IL-3 and survival kinetics after factor removal. All other cells tested behave like FDC-P1, suggesting that the metabolism of IL-3 by FDC-P2 is exceptional. Our study indicates that transient proliferation of committed progenitor cells in the absence of added factors is apparently not due to a stable pool of internalized CSF but merely represents an intrinsic capability of these cells.

Animals↗
PubMed1990

Independent prognostic value of ploidy in colorectal cancer. A prospective study using image cytometry.

In a prospective study, the DNA content of Feulgen-stained nuclei obtained from fresh samples of 211 colorectal adenocarcinomas was evaluated by means of image analysis. The DNA histogram classification took into account aneuploidy and S-phase fraction for diploid cases. No significant relationship was found between ploidy and sex, age, preoperative carcinoembryonic antigen (CEA), size of the tumor, histologic differentiation, or Dukes' stage. Aneuploidy was more frequently encountered in distal tumors. Preoperative CEA, histologic differentiation, Dukes' stage, and ploidy were individually associated with overall survival. In Dukes' A, B, and C tumors, patients with normal and elevated CEA had no significant difference in overall survival. A relationship was apparent between disease-free survival and site, histologic differentiation, Dukes' stage, and ploidy. Multivariate overall survival analysis did not reveal independent prognostic significance of ploidy when all Dukes' stages were considered. In contrast, Dukes' stage, differentiation, and ploidy were good indicators of higher risk of colorectal cancer-related death in patients undergoing curative surgery. Dukes' stage and ploidy were also indicators for recurrence. Thus, routine histopathologic characteristics should be used in combination with quantitative cytologic features for the definition of a relevant prognostic index in colorectal cancer.

Adenocarcinoma↗
PubMed1990

Human multi-drug-resistant cancer cells exhibit a high degree of selectivity for stereoisomers of verapamil and quinidine.

An in vitro cell proliferation assay was developed to measure the capacity of substances to overcome multi-drug resistance (MDR). The assay is a modification of the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) method. The inclusion of cell titration curves for each concentration of the resistance modifier (RM) allows the IC50 of the RM to be calculated and provides empirical correction of the cell survival curves for the effect of the RM when it is combined with a standard cytotoxic drug, vincristine. The resistance modification index (RMI) is defined as the ratio of the IC50 of vincristine obtained in control cultures divided by that measured in the presence of RM and is linearly related to the dose of RM. The RMI0.1, the RMI at a one-tenth the IC50 of the RM, provides a relative comparison between the activities of different RMs at non-toxic doses. The results obtained using the MDR cell line, KB-8-5, show that l-(-)-verapamil is approximately 4 times more active than d-(+)-verapamil in modifying MDR. The racemic mixture has an intermediate activity. A similar comparison between the epimers quinidine and quinine shows that, at equimolar doses, quinine has a higher RMI but, because it is more toxic, the RMI0.1 is about one-half of that of quinidine. These results demonstrate the importance of comparing the resistance-modifying activities of different compounds at doses relative to their own toxicity.

Cell Line↗
PubMed1989

Epidermal growth factor responsiveness of a new human neuroblastoma cell line.

A human neuroblastoma cell line, CA-2E, has been established from a bone-marrow aspirate of a 16-month-old boy with progressive disease. The karyotype and antigen phenotype of the cells correspond to those of a neuroblastoma. This cell line grows well in liquid cultures supplemented with 5% fetal calf serum; conversely, colony formation in semi-solid medium by cells from early passages is dependent upon exogenous EGF. With time in continuous culture, the cloning efficiency in the absence of EGF increases, but the line remains sensitive to EGF, as evidenced by an enhancement of the number and size of colonies. A relative dependence upon EGF in liquid cultures has also been clearly demonstrated by limiting the concentration of serum. Long-term (over 2 weeks) treatment with EGF results in a decreased rate of proliferation, a decreased proportion of clonogenic cells, and the appearance of flat, epithelial-type cells. In some experiments, EGF also has a remarkable effect in inducing neurite outgrowth and process branching. Our results suggest that EGF may have both proliferation- and differentiation-inducing effects on this neuroblastoma cell line. We have also shown that EGF induces increased proliferation in 7 out of 8 other human neuroblastoma cell lines. Functional response of neuroblastoma cells to EGF appears to be a general phenomenon which may be related to a block in the normal maturation pathway of the neural crest cells from which this tumor originates.

Animals↗
PubMed1989

Proliferation of single hemopoietic progenitor cells in the absence of colony-stimulating factors and serum.

The proliferative capacity of partially purified populations of mouse bone marrow progenitor cells has been examined in serum-free cultures in the presence and absence of colony-stimulating factors (CSFs). The half-life of progenitor cells was approximately 25 h in the absence of factor, regardless of whether the factor used to rescue the cells was granulocyte-macrophage CSF (GM-CSF), granulocyte CSF (G-CSF), or interleukin 3 (Il-3). During 4 days of culture without stimulator, the cells proliferated with a doubling time of about 24 h, in contrast to 8-12 h in response to exogenous CSF. To determine if this apparent CSF-independent proliferation was due to production of factors in situ, colony and single-cell transfer experiments were performed in serum-free media. The results showed that isolated hemopoietic progenitor cells can respond to GM-CSF, G-CSF, or Il-3 to give clones of greater than 100 mature granulocytes and macrophages after 4 days. Some transferred cells divided up to four times in the absence of factor, giving rise primarily to neutrophils. Even after an acidic wash, single cells were able to divide up to three times without stimulator, indicating that surface-bound factor can account for at most one division.

Animals↗
PubMed1988

Aggravation of experimental cutaneous leishmaniasis in mice by administration of interleukin 3.

Previous studies from our laboratory have shown that some in vitro maintained Leishmania major-specific L3T4+ T cells were capable of exacerbating cutaneous leishmaniasis after adoptive transfer to normal syngeneic mice. Results presented in this report show that these cells released substantial amounts of interleukin 3 (IL 3) and granulocyte-macrophage colony-stimulating factors after specific stimulation in vitro. In order to assess the involvement of such lymphokines in the exacerbation of cutaneous leishmaniasis by these L3T4+ T cells, the effect of the administration of important doses of IL 3 on the course of infection with L. major was investigated. The treatment of genetically susceptible BALB/c mice with IL 3 resulted in an enhancement of the size of lesions and favored the multiplication of parasites at anatomical sites distant from the primary lesion. Although IL 3 did not modify the development of lesions in genetically resistant CBA mice, this lymphokine promoted the growth of Leishmania in lymph node draining the lesion. Finally, the addition of IL 3 to macrophages parasitized in vitro enhanced the survival of intracellular Leishmania major.

Animals↗
PubMed1988

Inhibition of hemopoiesis in murine marrow cell cultures by recombinant murine tumor necrosis factor alpha: evidence for long-term effects on stromal cells.

The addition of recombinant murine tumor necrosis factor alpha (rmTNF-alpha) to serum-free methylcellulose cultures inhibited macrophage colony formation stimulated by purified colony stimulating factor-1 (CSF-1), recombinant granulocyte-macrophage-CSF (rmGM-CSF), and recombinant interleukin 3 (rmIl-3). The concentration of rmTNF-alpha inhibiting colony formation by 50% (IC50) was between 2 and 20 ng/ml. Erythroid colony formation in cultures with erythropoietin (EPO) alone or EPO, rmIl-3, and rmGM-CSF in combination were reduced to a much lesser extent. In established long-term marrow cultures (LTMC), addition of 20 and 200 ng/ml of rmTNF-alpha resulted in release of cells from the adherent layer during the first week. Treatment of cultures with rmTNF-alpha for 4 consecutive weeks led to prolonged inhibition of cell production lasting up to 8 weeks after cessation of treatment. One day after addition of a low dose of TNF (2 ng/ml), "fat" cells were no longer observed in the adherent layer. Our results indicate that TNF inhibition of hemopoiesis occurs both at the progenitor cell and stromal cell levels.

Animals↗
PubMed1988

The roles of granulocyte-macrophage colony-stimulating factor and interleukin 3 in stromal cell-mediated hemopoiesis in vivo.

Adherent cells from murine long-term marrow cultures (LTMC) were examined for presence of mRNA for granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin 3 (Il-3). Six hours after medium replacement, GM-CSF mRNA was detected but was no longer detectable 24 h after feeding; Il-3 mRNA was not detected at any time. Neutralizing antibodies against these factors had no effect on hemopoiesis. Exogenous Il-3 increased cell production, notably mature erythroid progenitors, whereas GM-CSF had little long-term effect even at high concentrations. Furthermore, GM-CSF appeared to be specifically removed from the medium, whereas virtually all of the Il-3 could be recovered under identical incubation conditions. These results show that Il-3 is not required for maintaining long-term hemopoiesis in vitro, whereas the precise role of GM-CSF in this system remains unclear.

Animals↗
PubMed1987

Colony formation in vitro as a prognostic indicator for primary breast cancer.

Cells from some, but not all, tumor biopsy samples form colonies when cultured in semi-solid media. The possibility that colony formation by progenitor cells in these tumors may reflect a more "aggressive" phenotype bearing clinical implications was examined in a series of 61 patients with primary breast cancer. Tumor cells from 32 samples formed colonies in vitro. There was no correlation between colony formation and any of the standard clinical parameters such as tumor size, nodal status, metastatic spread, or hormone receptor levels. Eighteen patients had inflammatory, locally advanced and/or detectable metastatic breast cancer at the time of surgery. Sixteen of these patients have progressed and 15 have died, with no relationship between colony formation and survival. For the 43 remaining patients, 23 had a tissue sample that gave rise to colonies in vitro; 14 of these have relapsed, with a median relapse-free survival (RFS) of 37.6 months, and eight have died with a median survival time of 46.8 months. This is compared with four relapses (median RFS not reached, P = .0043, Peto-Pike), and four deaths (median not reached, P = .1175) in the group without growth of the tumor specimen. These results indicate that colony formation is an independent prognostic parameter for breast cancer, which may be useful for selecting patients who would benefit from more intensive therapy.

Adult↗
PubMed1986

[Comparative prognostic study of the in vitro and in vivo development of colorectal tumors. Preliminary communication].

In vitro clonal growth of tumour cells may reflect biological properties of cancer and thus have prognostic value. This study seeks to establish correlations between the clinical outcome in patients after surgery for colorectal cancer and clonal growth of their tumours. History, status and follow-up data are collected. Tumour samples taken at operation under sterile conditions are plated immediately in our methylcellulose clonal assay system. Out of 65 consecutive samples, 3 did not yield sufficient cells for culture. Thirty-four (55%) grew more than 0.3 colonies/10(5) cells seeded; cloning efficiency was greater than 10 colonies/10(5) cells in 19. The 28 (45%) failures included 3 benign polyps cultured; 7 samples had visible bacterial or fungal contamination. The other 18 negative cultures may be due to cytotoxicity of the antibiotics or heterogeneity of tumour cells. These preliminary results show that colorectal cancers grow well in vitro in the absence of restricting factors, but they do not confirm the hypothesis that proliferative potential and differentiation are opposing processes. Location of the tumour may play a role, since best growth was seen in tumours of the caecum and terminal colon.

Cell Division↗
PubMed1986

Granulocyte-macrophage colony formation in serum-free culture: effects of purified colony-stimulating factors and modulation by hydrocortisone.

The effects of three purified colony-stimulating factors (CSFs) with different specificities for the granulocyte (G) and macrophage (M) lineages (G-CSF, CSF-1 and GM-CSF) were studied in a serum-free clonal assay system. The results were compared with those obtained in similar cultures containing fetal calf serum (FCS). Total clone (greater than or equal to 10 cells) and colony (greater than or equal to 50 cells) numbers were enhanced by FCS under most conditions. However, the extent of enhancement was highly dependent on the concentration and type of CSF. In some instances, FCS also altered the proportions of G, M, and mixed GM clones induced by the CSFs. In cultures stimulated with GM-CSF, enhancement by FCS was significant only at low CSF concentrations, primarily due to increased numbers of M clones. In contrast, clonal growth was increased by FCS only at high concentrations of CSF-1. Clone and colony numbers induced by G-CSF were greatly increased in cultures with FCS at all CSF concentrations tested. Virtually all clones developing in serum-free medium with G-CSF were pure G, whereas, M and GM clones were usually present in serum-containing cultures with high doses of G-CSF. The effects of hydrocortisone (HC) were also examined in these experiments. Like modulation by FCS, modulation of clonal growth by HC depended on the CSF used as stimulus, having no effect in cultures with G-CSF, inhibitory effects with CSF-1, and variable effects with GM-CSF related to CSF concentration.

Animals↗
PubMed1986

Stimulation of hematopoiesis in vivo by recombinant bacterial murine interleukin 3.

Mouse interleukin 3 (IL-3) cDNA was cloned into a plasmid construction, allowing the synthesis of very high quantities of IL-3 in Escherichia coli. The recombinant (r) IL-3, purified to homogeneity, was active in vitro on the proliferation and differentiation of various hematopoietic progenitor cells at 1 pM. To maintain detectable blood levels of IL-3, osmotic pumps containing rIL-3 or control solutions were placed under the skin of normal and irradiated C3H/HeJ and (BALB X B10) F1 mice. The effect of IL-3 on hematopoietic progenitor cell numbers in spleen and bone marrow was evaluated 3 and 7 days later by using an in vitro clonal assay. The results demonstrated the following: (i) Doses of IL-3 infused at the rate of 2.5-5 ng per g of body weight per hr were sufficient to increase the numbers of hematopoietic progenitors in normal mice by at least 2-fold within 3 days. (ii) In mice with progenitor cell levels depressed by sublethal irradiation, 7-day treatment with IL-3 resulted in a 10-fold increase to near normal levels. (iii) The erythroid and myeloid lineages appeared to be enhanced to the same extent. (iv) Enhancement of hematopoiesis occurred primarily in spleen, but hematopoietic foci were also evident in the liver; in contrast, total cell and progenitor cell numbers were decreased in the bone marrow.

Animals↗
PubMed1985

Recombinant murine GM-CSF from E. coli has biological activity and is neutralized by a specific antiserum.

We report the production and characterization of a mouse granulocyte-macrophage colony stimulating factor (mGM-CSF) made in Escherichia coli. The synthesis of mGM-CSF was directed by a plasmid containing a gene isolated from the EL-4 cell line. After induction of expression and accumulation of the protein in E. coli, mGM-CSF accounted for 10% of total cellular protein. This recombinant mGM-CSF was purified to 90% homogeneity by chaotrope extraction and gel filtration. Recombinant mGM-CSF, like the native molecule, stimulates the growth of granulocyte and macrophage colonies in serum-free cultures of mouse bone marrow cells. Antibodies raised against recombinant mGM-CSF not only reacted with the recombinant protein but also neutralized the biological activity of both native and recombinant mGM-CSF. These results indicate that the functional structure of the recombinant protein is similar to that of native mGM-CSF.

Amino Acid Sequence↗
PubMed1985

Non-linearity of colony formation by human tumour cells from biopsy samples.

The relationship between colony numbers and concentration of cells plated is an important parameter of clonogenic assay systems. The cloning efficiency for an ideal sample should be independent of cell concentration, thus giving a straight line through the origin when colony numbers are plotted against cell concentration. A simple statistical method has been developed to test if this is the case for individual tumour samples. Colony data from 51 freshly obtained tumour samples, which had sufficient cells to plate 3 or more dilutions and gave at least 20 colonies per plate at one or more of the dilutions, were tested. The results indicated that colony formation was linear for 27 (53%) of the samples. The remaining 24 samples could be classified into 2 groups: type I, in which cloning efficiencies increased with increasing cell concentration and type II, which had reduced cloning efficiencies at high cell concentrations. Fifteen (29%) of the samples had type I non-linearity and 9 (18%) exhibited non-linearity of type II. These findings indicate that the relationship between colonies and cells plated should be examined for each biopsy sample particularly in each experiment where the effects of cytotoxic drugs are tested.

Breast Neoplasms↗