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M P Scheid

Publications and source records attributed to M P Scheid.

At least 19 recordsLinked to original sources
PubMed1995

Role of phosphatidylinositol 3-OH-kinase activity in the inhibition of apoptosis in haemopoietic cells: phosphatidylinositol 3-OH-kinase inhibitors reveal a difference in signalling between interleukin-3 and granulocyte-macrophage colony stimulating factor.

Apoptosis of haemopoietic cells can be inhibited by various cytokines, but the specific signalling pathways involved are not well defined. Interleukin (IL)-4 has unique actions since it is unable to activate p21ras or mitogen-activated protein kinase, but can activate PtdIns 3-OH-kinase (PtdIns 3-kinase), the latter effect being shared with most other cytokines. In many cases, IL-4 is able to maintain cell viability by inhibiting apoptosis, but is unable to stimulate continuous proliferation of cells. This led us to investigate the role of PtdIns 3-kinase in inhibition of apoptosis. Two potent inhibitors of PtdIns 3-kinase, wortmannin and LY294002, rapidly induced apoptosis in cells incubated in the presence of IL-4, at concentrations consistent with their ability to inhibit PtdIns 3-kinase activity in whole cells. Interestingly, the same effect was seen in cells in the presence of IL-3 and Steel factor, while cells incubated in the presence of granulocyte-macrophage colony stimulating factor, and to a lesser extent, IL-5 could bypass the effect of wortmannin or LY294002. Therefore, this study suggests that PtdIns 3-kinase activity provides an important, although not a unique signal, required to inhibit apoptosis in haemopoietic cells.

Androstadienes↗
PubMed1995

A single-step purification of biologically active recombinant human interleukin-5 from a baculovirus expression system.

Recombinant human interleukin-5 (rhIL-5) was expressed in baculovirus-infected insect cells and purified to homogeneity from the culture medium in a single chromatographic step. Beginning with a cDNA encoding the full-length precursor form of human IL-5, including the authentic secretory leader sequence, recombinant baculovirus-infected insect cells expressed high levels of rhIL-5 (5-15 mg/liter culture) of which > 90% was processed to the mature form and secreted into the culture medium. After removing cells by centrifugation, rhIL-5 was purified by first adjusting the culture medium to the calculated pI value of mature IL-5 (pI 7.44) and then passing the conditioned medium through tandem linked anion- and cation-exchange columns. The resulting pass-through fraction contained the rhIL-5 and was devoid of contaminating proteins. An optional hydrophobic-interaction chromatography step effectively concentrated the pure homodimeric N-glycosylated rhIL-5 with a high overall yield (> 90%). N-terminal amino acid sequence determination indicated that cleavage of the human IL-5 leader sequence in insect cells occurred between Ala19 and Ile20. Recombinant human IL-5 prepared by this procedure bound to the high-affinity IL-5 receptor present on an eosinophilic leukemia cell line and elicited a proliferative response in the IL-5-dependent murine B-cell line BCL1. This rapid and simple procedure for the expression and purification of mature rhIL-5 should therefore enable studies requiring large amounts of this cytokine.

Amino Acid Sequence↗
PubMed1994

Induction, characterization, and functional coupling of the high affinity chemokine receptor for RANTES and macrophage inflammatory protein-1 alpha upon differentiation of an eosinophilic HL-60 cell line.

Eosinophilic differentiation of a pro-eosinophilic HL-60 cell line resulted in the induction of a high affinity RANTES/macrophage inflammatory protein-1 alpha receptor. The induced receptor is biochemically indistinguishable in RANTES equilibrium-binding studies from the monocytic receptor expressed on THP-1 cell membranes. Continued expression of the receptor requires the continuous presence of the inducing stimulus, and receptor site number declines without a loss of binding affinity with a t1/2 of 11.5 h on withdrawal of the inducing stimulus. The induced receptor is capable of three physiologic measures of receptor coupling, namely, ligand-induced Ca2+ fluxes, priming of the respiratory burst, and chemotaxis. Dose-dependent Ca2+ fluxes were elicited upon increasing concentrations of RANTES and MIP-1 alpha whereas no response was measured upon addition of MIP-1 beta or MCP-1. In addition, desensitization studies demonstrated that previous exposure to either RANTES or MIP-1 alpha almost completely inhibits a Ca2+ flux upon subsequent exposure to either ligand. Priming of the respiratory burst to PMA in differentiated cells by human rRANTES was more effective than priming by IL-5 or granulocyte-macrophage-CSF, whereas undifferentiated cells failed to secrete superoxide anion. In addition, differentiated cells underwent chemotaxis in response to RANTES. This provides the first evidence for the induction of a C-C chemokine receptor upon eosinophilic differentiation of a leukocyte cell line, and is in keeping with the demonstrated ability of human RANTES to induce the rapid formation of eosinophilic inflammatory sites.

Butyrates↗
PubMed1989

Reticulum cell sarcomas of SJL mice have rearranged immunoglobulin heavy and light chain genes.

The immunoglobulin (Ig) heavy (H) and light (L) chain gene rearrangements of the high incidence SJL lymphomas (reticulum cell sarcoma, RCS) have been analyzed. Both primary and transplanted RCS show rearrangements of H and kappa L chains, demonstrating that these tumors are of B cell origin. These data are consistent with previous results indicating that these tumors are a mouse model for follicular lymphoma. A long-term transplanted line and the in vitro line derived from it, cRCS-X, have a single rearranged JH-C gamma 2a fragment and one rearranged C alpha gene fragment which does not hybridize with a probe for the JH gene segments. These cell lines also have two rearranged J kappa-C kappa fragments. Primary tumors and early passages are more heterogeneous with respect to Ig gene rearrangements, possibly because more than one B cell clone is present. Although no synthesis of IgG2a, or of any Ig, could be detected by the in vitro cRCS-X cells, these cells contain abundant poly(A)+ RNA that hybridize with gamma 2a and kappa probes as well as lesser amounts of alpha and epsilon RNA. None of these H chain RNA hybridized with probes for the JH gene segments. The epsilon and alpha RNA are the same size as transcripts of germ-line CH genes which have been identified in other systems. However, the gamma 2a RNA are smaller than previously described germ-line C gamma 2a RNA and appear to be transcribed from aberrantly rearranged JH-C gamma 2a genes.

Animals↗
PubMed1986

Development of T-cell function in relation to T-cell set diversification in nu/nu mice.

The differentiation pattern of splenic T-cell populations in germ- and pathogen-free nu/nu mice, as compared to nu/+ littermates, is characterized by two abnormal features: the expression of TL determinants on peripheral T cells and the delayed onset of their differentiation from the predominant Lyt-123:TL+ set into TL- cells of Lyt-1+ and Lyt-123+ phenotype, which, in these mice, does not occur until 10 weeks of age. We report here that the delayed onset of mitogen- or alloantigen-induced interleukin-2 synthesis and T-cell proliferation as well as the development of cytotoxic T-lymphocyte activity of enriched T-cell populations is strictly correlated with the time point of T-cell subset diversification in nu/nu mice and depends in particular on the presence of the Lyt-1 (TL-:Lyt-2-) T-cell set which is lacking in splenic T-cell populations of germ-free young nu/nu mice.

Age Factors↗
PubMed1985

Regulatory influence of thymopentin on splenic T cell sets of thymectomized and aged mice.

Thymectomy of mice aged 6-8 weeks causes a disproportion of splenic T cell sets, the Ly123 set being relatively decreased and the Ly23 set relatively increased (Ly123 decrease: Ly23 increase). A similar disproportion of splenic T cell sets was found to occur spontaneously with advancing age (12-18 months). By PA-SRBC assay, the total number of splenic Lyt+ cells is not appreciably reduced by thymectomy or by aging, but the Thy-1+ cell count falls by about 40% according to both PA-SRBC and cytotoxicity assays. Thus there is an increase in the number of Lyt+ cells expressing sub-threshold amounts of Thy-1. The following observations show that thymopentin (TP-5), a synthetic pentapeptide analogue of thymopoietin, counteracts these changes in thymectomized and aged mice. As reported previously, the capacity of C3H/HeJ female mice to reject C3H/HeJ male skin was raised by thymectomy, or with age, and treatment with TP-5 substantially normalized the rejection response. Here we correlate these findings with changes in the profile of splenic T cell sets. The splenic T cell set profile of thymectomized B6-Tlaa male and female mice was essentially restored by TP-5. The Ly123 decrease: Ly123 increase change caused by thymectomy was not associated with obviously altered proportions of Qa-1+ and Qa-1- subsets. Treatment of aged mice with TP-5 also prevented the onset of changes in splenic T cell sets that occur spontaneously with age. Thus thymectomy and aging give rise to disproportions of splenic T cell sets, and in C3H female mice to a heightened capacity for male skin rejection, both effects being largely abrogated by the TP-5 derivative of thymopoietin.

Aging↗
PubMed1985

Phenotypic variation in clonal Abelson virus lymphoma cells.

Two clonal A-MuLV lymphoma cell lines have the capacity to generate phenotypic variants when grown in vivo as ascites tumors. Variant lines differed from parental lymphoma cells in their expression of enzymatic or cell surface differentiation markers. Parental lines expressed the B220 and Lyb-2 glycoproteins characteristic of pre-B cells and bound B220-specific monoclonal antibodies such as 14.8. The parental cells expressed low levels of TdT activity but did not synthesize detectable mu-heavy chain, a cellular phenotype that may correspond to lymphoid progenitor cells. Three classes of phenotypic variants were recovered from the Thy-1- parental lines: 1) 14.8+, Lyt-1+, Thy-1- cells; 2) 14.8 +/-, Lyt-1+, Thy-1+ cells, and 3) 14.8-, Lyt-1+, Thy-1+ cells. Cell cloning experiments indicated that Thy-1+ variant cells can be recovered within 14 days of in vivo inoculation as a minor proportion (1/10(6] of the tumor cell population and subsequently become the predominant tumor cell population. These clonal tumor lines provide a model for the study of cellular and molecular alterations that occur during neoplastic differentiation and progression in the lymphoid system.

Abelson murine leukemia virus↗
PubMed1984

Contrasting biological activities of thymopoietin and splenin, two closely related polypeptide products of thymus and spleen.

Thymopoietin, a 49 amino acid polypeptide hormone of the thymus discovered by its effect on neuromuscular transmission, was later shown to induce T-cell differentiation and to affect immunoregulatory balance. A radioimmunoassay for thymopoietin revealed a crossreaction with a product found in spleen and lymph node but not other tissues. This product, named splenin, differs from thymopoietin only in position 34, aspartic acid for bovine thymopoietin and glutamic acid for bovine splenin. Synthetic pentapeptides corresponding to residues 32-36, called thymopentin and splenopentin, reproduce biological activities of thymopoietin and splenin, respectively. Thus thymopoietin and thymopentin affect neuromuscular transmission and induce the phenotypic differentiation of T precursor cells in vitro while inhibiting phenotypic differentiation of B cells. Splenin and splenopentin, in contrast, do not affect neuromuscular transmission, and they induce both T- and B-cell precursors.

Amino Acid Sequence↗
PubMed1983

Characterization of interleukin 2-dependent cytotoxic T-cell clones. IV. Production of alpha, beta and gamma interferons and interleukin 2 by Lyt-2+ T cells.

The production of alpha, beta and gamma interferons (IFN) and interleukin 2 (IL-2) by Lyt-2+-dependent cytotoxic T-cell lines/clones was investigated. Cloned and uncloned T-cell lines specific for H-2Dd or the unique RL male 1 leukemia antigen were studied. After infection with Sendai virus (SV) or Newcastle disease virus (NDV) all cell lines produced IFN-alpha and -beta. Induction of IFN-gamma was attempted with the mitogens Con A, PHA, PWM, SEA, and SEB, with poly(I:C), with antibodies Lyt-1.2, -2.2, and Thy-1.2, or with the target cells Meth A (H-2Dd+) and RL male 1. All mitogens were effective inducers. However, the antibodies and poly(I:C) were not. One uncloned RL male 1-specific cell line CTLL-RP, produced IFN-gamma after induction with RL male 1. Production of IFN-alpha, beta depended on IL-2, whereas production of IFN-gamma did not, although addition of highly purified IL-2 increased IFN-gamma production even in the absence of other inducers. Crude IL-2 inhibited the production of IFN-gamma but not IFN-alpha, beta. In response to mitogens, some T-cell clones also produced IL-2. The results demonstrate that Lyt-2+ cells can produce a broad spectrum of lymphokine activities after appropriate stimulation. Their availability now affords us the opportunity to study the regulation of lymphokine production at the clonal level.

Animals↗
PubMed1983

Characterization of IL-2-dependent cytotoxic T-cell clones. II. Cell-surface phenotypes, histochemical and ultrastructural properties.

This report examines the histochemical staining patterns, ultrastructure, and cell-surface phenotypes of six antigen-specific T-cell clones. Histochemical analyses indicated that all cell lines expressed alpha-napthyl butyrate esterase characteristic of the monocytic isoenzyme, intense napthol AS-D chloroacetate reactivity characteristic of granulocytes, and were negative for leucoperoxidase, alkaline phosphatase, and Sudan black. Only one clone stained weakly for acid phosphatase. The esterase staining patterns became evident in newly established cell lines after growth for only 1 week in interleukin 2-conditioned medium. Ultrastructurally, the outstanding feature was numerous membrane-bound granules containing a complex-appearing globular material. The cell-surface phenotypes of the lines as determined by protein A-sheep red blood cell rosetting and indirect immunofluorescence was Ly-5+, T200+, Qa-5-, MAC-1-, and Lyt-1+,2,3- for the helper line and Lyt-1-2,3+ for the five cytotoxic lines. By quantitative absorption analyses, low levels of Lyt-1 antigens were detected on all examined cytotoxic lines. The results strengthen the view that long-term T-cell lines can retain normal T-cell characteristics while also expressing markers that are either absent or in undetectable levels on uncultured T lymphocytes. The presence of the esterases may be associated with an expanded functional role in the T-cell lines.

Animals↗
PubMed1983

Suppression of T cell cytotoxicity by nude mouse spleen cells: reversal by monosaccharides and interleukin 2.

The effects of monosaccharides on the suppression of cytotoxic T cell generation by spleen cells from nu/nu mice were examined. Suppression of the B6 anti-BALB/c response and the B6 anti-C3H response was reversed by alpha-methyl-D-galactoside (alpha MG) but not other sugars, including beta MG. Suppression was associated with a decrease in the level of IL 2, which suggests competition; this decrease was also reversed by alpha MG.

Animals↗
PubMed1981

Structural features and selective expression of three Ly-5+ cell-surface molecules.

Conventional Ly-5 alloantisera precipitate cell-surface molecules of three sizes: 200K, 205K, and 220K. In SDS-PAGE, the rat monoclonal antibody 74/8' precipitates the same three molecules from both Ly-5.1 and Ly-5.2 cells. Cleveland mapping of the three molecules, precipitated by reaction of conventional Ly-5 alloantisera or 74/8' monoclonal antibody with lysates of 125I-labeled cells, disclosed no differences among the three molecular forms, but markedly distinguished all three Ly-5.1 molecules from all three Ly-5.2 molecules. Each of the three molecular forms can be expressed independently of the other two by cloned culture lines of Ly-5+ cells of different hematopoietic lineage. All of the seven cloned lines tested expressed only one form. However, two of the seven uncloned culture lines tested, plasmacytoma MOPC-70A and the X.1 putative macrophage line which originated from an SJL tumor, yielded both 200K and 205K forms.

Animals↗
PubMed1981

Effect of the TP5 analogue of thymopoietin on the rejection of male skin by aged and thymectomized female mice.

Although young adult C3H/HeJ (C3H) females do not reject C3H male skin grafts, C3H females older than 1 year commonly do so, as also do many thymectomized, young adult C3H females. Therapy with TP5, a synthetic pentapeptide analogue of thymopoietin which has biological properties of the parent molecule, substantially reduced the capacity of aged C3H females and of thymectomized, young C3H females to reject C3H male skin.

Aging↗
PubMed1981

Immunological studies of mouse decidual cells. II. Studies of cells in artificially induced decidua.

Cells from artificially induced decidual tissue (deciduoma) in the mouse were examined for Thy-1 surface antigen and receptors for the Fc portion of immunoglobulin G (FcR) and compared with cells of the normal decidua from 6 to day 9 of pregnancy. It was shown that (1) Thy-1 antigen is present on the same proportion of cells in decidua and deciduoma on day 6 and day 7, (2) FcR-bearing cells can be detected in similar numbers on day 6 and day 7 but this does not increase on day 8 in deciduoma as it does in decidua, and (3) progesterone treatment after induction of decidualization allowed further increase of FcR-bearing cells in deciduoma. These results present further evidence of the similarity between deciduoma and decidua in the mouse. They indicate that these two membrane markers are present in the early decidua, regardless of the presence of an embryo, and suggest that progesterone may play a part in the increase of FcR-bearing cells in the decidua during pregnancy.

Animals↗
PubMed1981

Cellular interactions affecting the maturation of murine B lymphocyte precursors in vitro.

Clonable B cells arose in liquid cultures of slg+ cell-depleted bone marrow suspensions, and a majority of these derived from rapidly sedimenting (presumably large) precursors. Such immature (pre-B) cells were not demonstrable in adult lymph nodes. The development of functional B cells in vitro depended on adherent cells present in adult bone marrow or peritoneal exudates and in the presence of optimal numbers of these accessory cells; not FCS or 2-ME or LPS were essential for B cell maturation. In contrast, the ability of fetal liver cell suspensions to generate B cells in culture was not affected by the removal of adherent cells or addition of macrophages, and LPS consistently reduced numbers of colony-forming B cells recovered. A type of nonadherent cell present in the bone marrow of adult CBA/N mice dramatically enhanced B cell maturation in normal CBA/H-T6T6 fetal liver cell cultures. These results indicate that 2 types of accessory cells may augment transition of pre-B cells to functional B cells in culture. One is adherent and can be replaced by macrophages, whereas the other is nonadherent and is present in limiting numbers in embryos. When fetal liver cultures were incubated with optimal numbers of adult accessory cells, the rate of emergence of B cells was dependent on the gestational age of the donor embryo. This suggests that B cell formation in utero may be limited by the availability of B cell precursors as well as nonadherent accessory cells. The possibility exists that fetal pre-B cells typify intermediates in the B lineage which are infrequent in adult marrow. Alternatively, B cells may be produced in adult life through processes that do not precisely recapitulate embryonic events.

Animals↗