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Biomedical subjects

E Lundgren

Publications and source records attributed to E Lundgren.

At least 145 records · Page 8Linked to original sources

The augmentation of human natural killer cell activity by interferon-gamma is not associated with the induction of the interferon-alpha-inducible proteins.

Treatment of partly purified large granular lymphocytes (LGL) with either IFN-alpha or IFN-gamma for 2 hr augmented their NK cell activity. This augmentation was completely inhibited by the addition of 10 micrograms/ml of cycloheximide. In contrast, when the effects of IFN-gamma on the synthesis of specific proteins in these cells was directly studied by use of two-dimensional gel electrophoresis, we found that IFN-gamma was unable to induce any of the earlier detected, IFN-alpha/IFN-beta-inducible proteins within 18 hr of incubation. No additional, IFN-gamma-induced proteins were detected in either the partly purified LGL or purified T cells. In contrast, the effects of the two factors were comparable in the glioma cell line 251 MG. This shows i) that the effects of IFN-alpha and IFN-gamma are dependent on the responder cell type, ii) that there exists at least one mechanism that can augment NK cell activity that is not dependent on the increased synthesis of the IFN-alpha-inducible proteins, and iii) that either the nine IFN-alpha-inducible proteins are not involved in any leukocyte function that is augmentable by both IFN-alpha and IFN-gamma, or that the two factors exert their actions in leukocyte through different mechanisms.

Adjuvants, Immunologic↗

Enhancement of immunoglobulin secretion by the lymphokine-like activity of a Mycoplasma arginini strain.

B lymphocytes, preactivated by lipopolysaccharide (LPS), could be triggered to growth by a strain of Mycoplasma arginini, while the level of immunoglobulin (Ig) secretion, quantitated as the number of plaque-forming cells (PFC), was low. The PFC response could be increased by the addition of conditioned media (CM) from lectin-activated spleen cells or T cell tumour EL-4 to the culture of preactivated B cells. CM did not by itself induce a significant amount of PFC in the cultures of LPS-preactivated B cells. The maturation enhancing activity was distinct from IL-2 and B cell growth factor as judged by gel exclusion chromatography.

Animals↗

Kinetics of styrene in workers from a plastics industry after controlled exposure: a comparison with subjects not previously exposed.

Eight male workers from a glass reinforced plastics industry were experimentally exposed for 2 hours to 2.84 mmol/m3 (296 mg/m3) styrene during light physical exercise (50 W). About 63% of the amount supplied (4.6 mmol styrene) was taken up in the body. The arterial blood concentration of styrene reached a relatively stable level of 15 mumol/l at the end of exposure which was about 70% of the blood concentration in a group of volunteers with no previous exposure to solvents. The apparent blood clearance was significantly higher in the occupationally exposed subjects 2.01/h X kg compared with 1.51/h X kg. Contrary to the relatively stable level of styrene at the end of exposure the concentration of non-conjugated styrene glycol increased throughout the exposure and reached about 3 mumol/l in both groups. Like styrene, the non-conjugated styrene glycol seemed to be eliminated faster from the occupationally exposed workers. The blood concentration of styrene-7,8-oxide was low and seldom exceeded the detection limit of 0.02 mumol/l. The results show that long term exposure in a glass reinforced plastics industry may facilitate the metabolism of styrene.

Adult↗

Biological monitoring of styrene metabolites in blood.

Ten men occupationally exposed to styrene in two glass-fiber reinforced plastics factories were studied during three consecutive workdays. The mean external exposure level was 99 mg/m3. The total pulmonary uptake of styrene was estimated from measurements of the styrene concentration in inspired air, the pulmonary ventilation, and the relative uptake. A gas chromatographic method based on electron capture detection was used to quantify styrene glycol, as well as styrene-7,8-oxide, in blood. The concentration of styrene glycol appeared to be linearly related to the preceding uptake of styrene. When the uptake during 5 h immediately before the blood sampling was considered, the correlation coefficient (r) obtained the value of 0.90. The concentration of styrene-7,8-oxide was at the detection limit of 0.02 mumol/l in most samples. A weaker correlation between the concentration of styrene in blood and the uptake during the hour immediately preceding the blood sampling was obtained (r = 0.71).

Adult↗

Adhesion of cardiac myocytes to extracellular matrix components.

The interaction of ECM components with adult cardiac myocytes is not well understood, but is of physiological importance. Most physiological studies are conducted on myocytes in suspension yet in vivo the cells are attached to each other and to the ECM. In this paper, we further define the interaction of isolated adult myocytes with the ECM substrates. Of interest is not only the short-term attachment of cells to ECM substrates but also the ability of ECM components to support the long-term maintenance of cardiac myocytes in cultures.

Animals↗

Evidence for IFN-beta heterogeneity in a substrain of Namalwa cells.

A substrain of Namalwa cells, denoted substrain B, was grown in fermentors up to the 100-L scale, and was induced with Sendai virus to produce interferon (IFN). The titer of the crude IFN varied extensively between different batches; part of the variation was caused by a differential expression of IFN-alpha and IFN-beta. More than 80% of the IFN activity was IFN-beta by several criteria. A two-step purification procedure was developed and the resulting preparation had a specific activity of approximately 10(6) U/mg protein. The IFN-beta type was found to be heterogeneous, and could be separated into several components, which probably represented post-translational modifications of one molecule.

Adsorption↗

Extracellular matrix components influence the survival of adult cardiac myocytes in vitro.

Calcium-tolerant myocytes were isolated from adult rat hearts by collagenase perfusion and plated on various substrates in serum-free medium and their adhesion to various extracellular matrix (ECM) components was determined. The myocytes attached readily to dishes coated with collagen type IV (C-IV), laminin (LN), and to fetal bovine serum (FBS) in a manner dependent on the concentration of the components. Substantially fewer myocytes adhered to dishes coated with fibronectin (FN) or to uncoated plastic dishes. Cells adhered equally well to dishes coated with C-IV, LN and FBS within 1-4 h. However, when examined after 2 weeks in culture it was found that only C-IV and LN could support survival of the attached myocytes, and when cultured on C-IV or LN the myocytes were spread and had formed a dense monolayer. The actin filaments had at this time reorganized linearly along the long axis of the cell and the myocytes contracted spontaneously. Rabbit antibodies were raised against myocyte membranes and their ability to inhibit attachment to ECM components was studied. Purified IgG inhibited attachment to C-IV, while having only a minor effect on attachment to LN. These data are compatible with the presence of a specific cell surface component(s) that interacts with ECM substrates and influences cell shape and possibly thereby influences cellular functions.

Animals↗

Differential expression of interferon genes in a substrain of Namalwa cells.

A substrain of Namalwa cells producing a high ratio of beta-interferon (IFN-beta) versus alpha-interferon (IFN-alpha) was investigated by constructing a cDNA library after induction with Sendai virus. The library was screened by two synthetic oligonucleotides, one specific for IFN-alpha and one complementary to both IFN-alpha and IFN-beta. Rescreening the library with two full-length cDNAs encoding IFN-alpha and IFN-beta, respectively, revealed that the frequency of the IFN-alpha and IFN-beta clones reflected the activities of IFN-alpha and IFN-beta obtained by functional assays. On the cDNA level, the dominating species was identical with the type of IFN-alpha A or IFN-alpha 2; however, one new type of cDNA also was found that was similar to the previously described IFN-alpha C. Only one type of cDNA was found encoding IFN-beta, although several IFN-beta proteins have been detected in the analyzed cell line.

Base Sequence↗

Mycoplasma mimicry of lymphokine activity in T-cell lines.

During screening of T-T hybridomas for B-cell-stimulating factors (BSF) we found in the conditioned medium of such a line (TUH-14) an activity that turned out to cosediment with a contaminating strain of Mycoplasma arginini on centrifugation. Since it only affected B blasts and only to a minor extent small cells and since it did not induce immunoglobulin secretion, the mycoplasma strain seemed to mimic a BSF. M. arginini was also found in two other T-T hybridoma producer lines but not in a clone of the EL-4 line. Three other M. arginini isolates were not stimulatory for B cells. The presented results point towards the importance of careful control for mycoplasma contamination in established cell lines.

Animals↗

Similar effects of treatment with alpha interferon on the protein synthesis of human large granular lymphocytes, T cells, and monocytes.

Preparations of human large granular lymphocytes (LGL), T cells, and monocytes (MC) were obtained through centrifugation on Percoll gradients and preparative E-rosetting. The different preparations contained more than 80% of the appropriate cell type, as judged by their ability to lyse 51Cr-labelled K562 cells, cell morphology, and the presence of cell surface structures recognized by the OKT3, OKT10, Leu 7 and OKM1 monoclonal antibodies. The protein synthesis is unstimulated and alpha interferon (IFN-alpha)-treated cells of the different types was studied by subjecting 35S-methionine-labelled cell extracts to two-dimensional gel electrophoresis. The general pattern of protein synthesis in LGL and T cells was virtually identical, whereas at least 7 major proteins were synthesized at a higher rate in monocytes. The effects of IFN-alpha on the protein synthesis of LGL and T cells were identical, IFN-alpha increasing the rate of synthesis of 9 proteins. These proteins were also expressed, but not always IFN-augmentable, in monocytes. No additional, cell-type associated, IFN-inducible proteins were found. This suggests that the augmenting effect of IFN-alpha on the cytotoxic capacity of LGL, T cells, and monocytes may be to affect common steps in their lytic machineries.

Antibodies, Monoclonal↗

A purge-and-trap method for the analysis of acetone in biological tissues.

A method for the detection and quantitation of acetone in biological tissues is described. The solvent was extracted with nitrogen gas from the biological specimen and adsorbed on a porous polymer (Porapak Q). The acetone content of the adsorbent was determined by conventional gas chromatographic analysis (FID) after heat desorption. The reliability of the method was tested both with in vitro- and in vivo-exposed specimens. A complete recovery was obtained after in vitro additions of acetone. The method error was about 9% as calculated from both in vitro and in vivo experiments. Acetone concentrations ranging between 17 nmol/g tissue in nonexposed animals and 1.8 mumol/g tissue in mice exposed to acetone vapor were determined.

Acetone↗

Recognition of extracellular matrix components by neonatal and adult cardiac myocytes.

Recognition of extracellular matrix (ECM) components by isolated cardiac myocytes from neonatal (4-5 days postpartum) and adult rats was determined by measuring cell attachment to substrates made of ECM components. The substrates were petri dishes coated with either fibronectin, laminin, native monomers of collagen types I, II, III, IV, and V, denatured collagen, or gels containing reconstituted collagen fibers. Adult myocytes attached efficiently to laminin and type IV collagen, weakly to fibronectin, but not at all to the other types of collagen. Neonatal myocytes attached well to all types of collagen and to fibronectin and laminin. Antibodies raised against surface membranes of neonatal myocytes, adult myocytes, or adult hepatocytes were assayed for their ability to inhibit cell attachment to the various ECM substrates. Antibodies against the surface of neonatal myocytes as well as antibodies against the hepatocyte cell surface inhibited the attachment of neonatal myocytes and hepatocytes to collagen but not to fibronectin. Antibodies against the adult myocyte cell surface did not inhibit the attachment of neonatal myocytes or hepatocytes to ECM components. These results indicate the presence of binding molecules on the surface of neonatal myocytes that are involved in the recognition of collagen at a time when collagen is being secreted and formed into a three-dimensional network that attaches to the cell surface of the myocytes. This recognition and adhesion to collagen occurs by a mechanism independent of fibronectin. The binding molecules for collagen could not be detected on normal adult myocytes isolated at a time when the formation of the collagen network has already been completed.

Aging↗

Isolation, characterization and adhesion of calcium-tolerant myocytes from the adult rat heart.

High yields of Ca2+ - stable myocytes were obtained by perfusion of adult rat heart with a buffered collagenase medium followed by mincing and three additional digestion periods. Release of lactate dehydrogenase, respiratory control, content of ATP and creatine phosphate, electrical stimulation and attachment to extracellular matrix components indicated that the sarcolemma of the isolated myocytes remained intact and that the cells maintained some of the most basic physiological functions. The myocytes maintained their rod-shape in a medium containing 2.5 mM of Ca2+ and their release of LDH was slow. Some of the myocytes were contracting spontaneously, at a low rate, in an abrupt end-to-end contraction. Other cells appeared quiescent but they were all able to respond to external electrical stimulus. The oxygen consumption was measured by a perifusion method. In different preparations the basal consumption was 14-26 nmol O2/min X 10(5) rod-shaped myocytes. Freshly isolated rod-shaped heart cells attached in 30 minutes to dishes coated with collagen type IV, laminin or fibronectin but did not attach to dishes coated with collagen type I or III or to collagen gels. Attachment occurred at the ends of the cells.

Adenosine Triphosphate↗

Novel cluster of alpha-interferon gene sequences in a placental cosmid DNA library.

A human cosmid library was constructed and probed with a human alpha interferon (IFN-alpha) cDNA clone. One clone giving a strong hybridizing signal was isolated and characterized. The cosmid DNA insert represents a section of the human genome containing three regions of IFN-alpha-like sequences. The DNA was characterized with restriction endonuclease mapping, thereby allowing comparison to similar linkage groups reported recently and determination of homologous regions on the known physical map. The three IFN-alpha-like sequences were analyzed by a partial sequence analysis. Mapping and sequence data establish this section as a not-yet-described cluster of IFN-alpha sequences in the human genome; however, a part of the section matches to some degree to a previously described genomic region. The region described here could represent genetic polymorphism or a duplicated segment.

Base Sequence↗

Interferon therapy in juvenile laryngeal papillomatosis.

Seventeen patients with severe juvenile laryngeal papillomatosis (12 in Stockholm and five in Umeå) were treated with exogenous leukocyte interferon (IFN-alpha) prepared in Helsinki and Umeå, respectively. Tumor progression occurred in all cases before treatment. During treatment tumor growth diminished in all cases, up to complete tumor disappearance. Of 17 patients, nine were cured and no longer are being treated, four exhibit no tumor growth but are still being treated, one has visible tumor but only slight growth, two still have active but diminished growth, and one, who has refused further treatment, is experiencing active tumor growth as before the start of interferon therapy. It is concluded that IFN-alpha therapy in a dosage of 3 X 10(6) units three times a week intramuscularly can arrest papilloma growth. Further trials are needed to optimize treatment.

Adolescent↗

Occurrence of styrene-7,8-oxide and styrene glycol in mouse after the administration of styrene.

Styrene-7,8-oxide and its hydrated product styrene glycol were determined in mouse tissues at different times (0.5-5 h) after the intraperitoneal administration of 7-[14C]-styrene (3.8 mmol/kg). In a study of the influence of dose on the metabolite pattern of styrene, mice were killed 2 h after a dose of 1.1, 2.3, 3.4, and 5.1 mmol/kg, respectively. The mouse tissues studied (blood, liver, kidney, lung, brain, subcutaneous adipose tissue) were isolated and extracted first with hexane to remove styrene and styrene-7,8-oxide and then with ethyl acetate to remove styrene glycol. beta-Glucuronidase was used to liberate conjugated styrene glycol. A gas-liquid chromatographic method based on the use of an electron capture detector (GLC-EC) was used to quantify styrene glycol, as well as styrene-7,8-oxide, after hydrolysis. In addition all homogenates and extracts were assayed by radioactivity counting. Styrene-7,8-oxide and styrene glycol reached maximum concentrations within 2 h. The highest levels of styrene-7,8-oxide were detected in the kidneys and subcutaneous adipose tissue, while the lungs showed the lowest levels. Styrene glycol was found in the highest concentrations in the kidneys, liver, blood, and lungs. The concentration of unmetabolized styrene increased exponentially at higher doses. There seemed to be a linear increase with the dose of styrene-7,8-oxide and styrene glycol in all the tissues studied. The more polar metabolites occurred at relatively lower levels in the liver and kidneys at higher doses. In a complementary study the epoxide hydratase inhibitor trichloropropene oxide was added to the removed tissues, and the hexane extracts were analyzed for styrene-7,8-oxide both by GLC-EC and mass spectrometry (GLC-MS).

Animals↗

Lymphokine-like activity of a strain of Mycoplasma arginini.

A Mycoplasma arginini strain, found to contaminate a T-T hybridoma designated TUH-14, was the source of a lymphokine-like activity with an ability to stimulate B-blasts to proliferate. Maturation to immunoglobulin secretion induced by the mycoplasma alone was low compared with induction by lipopolysaccharide (LPS), but could reach the same levels achieved with LPS by the addition of a B-cell maturation factor obtained from lectin-activated EL-4 thymoma cells. The mitogenic effect on B-cells was found only in the strain isolated from TUS-14; three other M. arginini strains were negative. Both mitogenic and nonmitogenic mycoplasma membrane preparations displayed higher affinity for B-cells than for T-blasts. However, membranes from the mitogenic strain, the TUH-14 isolate, bound better to activated blasts than to small resting cells, in contrast to the nonmitogenic strain G-230.

Adhesiveness↗

Interferon-induced cell cycle changes in human hematopoietic cell lines and fresh leukemic cells.

A panel of 26 human hematopoietic cell lines was tested for sensitivity to growth inhibition towards interferon-alpha (IFN-alpha) by estimating the effects on cell cycle phase distribution using flow cytometry analysis. The proportion of proliferating cells was assessed by studying the fractional increase of cells in mitosis during a 24-hr vinblastine block. Of 26 cell lines tested, 17 were sensitive to IFN-alpha, and the main cell cycle effect was accumulation in the G0-G1 phase. One Burkitt's lymphoma line, Namalwa, showed a decreased rate of progress through S without any G0/G1 accumulation. Three of the cell lines were also tested with IFN-beta and with IFN-alpha 2 produced by recombinant DNA technology. The latter IFN did not affect one of the cell lines; otherwise, the results were similar to those of IFN-alpha. Six of 16 clinical specimens from patients with hematopoietic neoplasias were IFN-sensitive, all displaying a G0-G1 block. Our results indicate that IFN sensitivity is an individually linked property unrelated to cell origin.

Cell Cycle↗