Analysis and Purification of Human Lymphoblastoid (Namalwa) Interferon Using a Monoclonal Antibody

Allen, Geoffrey; Fantes, K. H.; Burke, D. C.; Morser, John

doi:10.1099/0022-1317-63-1-207

Cited by 61 publications

(32 citation statements)

References 22 publications

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“…48) provided by Wellcome Research Laboratories, Beckenham, Kent, United Kingdom. Recombinant human and murine IFN-␥ (4 ϫ 10 7 and 1 ϫ 10 7 IU/mg of protein, respectively) was from Dr. G. Adolf, Ernst Boehringer Institut fü r Arzneimittelforschung, Vienna, Austria.…”

Section: Methodsmentioning

confidence: 99%

Janus Kinase-dependent Activation of Insulin Receptor Substrate 1 in Response to Interleukin-4, Oncostatin M, and the Interferons

Burfoot

Rogers

Watling

et al. 1997

Journal of Biological Chemistry

115

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In addition to a role in response to insulin and insulinlike growth factors, insulin receptor substrate 1 (IRS-1) is phosphorylated in response to IL-4, the interferons (IFNs) and oncostatin M (OSM). Here mutant cell lines lacking JAK1, JAK2, or Tyk2 were used to determine the role(s) of the Janus kinase (JAK) family of protein-tyrosine kinases in IRS-1 phophorylation. 32D cells, which do not express IRS proteins, were analyzed for any requirement for these proteins in response to the IFNs. For the mutant human fibrosarcoma cell lines, phosphorylation of IRS-1 through the insulin-like growth factor receptor is independent of JAK1, JAK2, or Tyk2. In contrast, phosphorylation of IRS-1 mediated by the Type I IFNs, IL-4, and OSM is JAK-dependent. For the ␣␤-IFNs, activation of IRS-1 is dependent on JAK1 and Tyk2, consistent with the interdependence of these kinases in the IFN-␣␤ response. Neither IRS-1 nor IRS-2 was detectably activated by IFN-␥. Consistent with this, activation of neither IRS proteins appears to be an absolute requirement for an antiproliferative or an antiviral response to the IFNs. For IL-4 and OSM phosphorylation of IRS-1 in the human fibrosarcoma cells is largely dependent on JAK1 but can also be mediated through Tyk2 or JAK2. Activation of phosphatidylinositol 3 -kinase in response to IL-4 and OSM, at least, was also JAK-dependent. The JAKs are, therefore, required not only for STAT activation but also for the activation, through a variety of different types of cytokine receptor, of an additional signaling pathway(s) through IRS-1 and phosphatidylinositol 3 -kinase.Receptors that have an intrinsic tyrosine kinase domain recruit and activate a variety of signal transducers. Insulin receptor substrate 1 (IRS-1) 1 is a cytosolic protein of ϳ180 kDa in mass that is tyrosyl-phosphorylated at multiple sites through activated insulin and insulin-like growth factor 1 (IGF-1) receptors (1, 2). An L(X) 4 NPXY(p)XSXP motif has been identified in the insulin receptor as being important for the recruitment of the IRS proteins. IRS-1 contains multiple YMXM and YXXM tyrosine motifs, which when phosphorylated can recruit Src homology 2 domain-containing proteins, including the p85␣ regulatory subunit of PI 3Ј-kinase, SHP-2, Grb-2, Crk, and Nck (3, 4). Phosphorylated IRS-1, therefore, can mediate a variety of responses including a mitogenic response and activation of PI 3Ј-kinase.Common themes involving the role of tyrosine kinases and proteins with Src homology 2 domains have emerged to describe broadly the mechanism of signal transduction by many growth factors and cytokines. Signal transduction pathways utilizing the JAK (Janus kinase) family of protein-tyrosine kinases and the STATs (signal transducers and activators of transcription) are activated in response to polypeptide ligands including many cytokines, some growth factors and the interferons (IFNs). STAT activation mediated through the JAKs occurs in receptor complexes at the cell membrane. There are four known mammalian JAKs: JAK1, JAK2, JAK3, and T...

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Section: Methodsmentioning

confidence: 99%

Janus Kinase-dependent Activation of Insulin Receptor Substrate 1 in Response to Interleukin-4, Oncostatin M, and the Interferons

Burfoot

Rogers

Watling

et al. 1997

Journal of Biological Chemistry

115

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“…Ceils, virus and conditions for induction of Namalwa cells were as described elsewhere (Allen et aL, 1982). Tunicamycin, a kind gift from Dr R. L. HamiU of Eli Lilley, U.S.A., was dissolved in dimethyl sulphoxide at 1 mg/ml and stored at -20 °C.…”

Section: The Effect Of Tunicamycin On Interferon-a Production In a Humanmentioning

confidence: 99%

“…RNA was extracted by the guanidinium isothiocyanate technique (Morser & Shuttleworth, 1981) and assayed by microinjection into oocytes (Colman & Morser, 1979). The procedures for purification and analysis of IFN-a were described by Allen et al (1982).…”

Section: The Effect Of Tunicamycin On Interferon-a Production In a Humanmentioning

confidence: 99%

“…Under these conditions we detected no inhibition of protein synthesis. IFN samples were collected and assayed as described by Allen et al (1982). RNA was extracted by the guanidinium isothiocyanate technique (Morser & Shuttleworth, 1981) and assayed by microinjection into oocytes (Colman & Morser, 1979).…”

Section: The Effect Of Tunicamycin On Interferon-a Production In a Humanmentioning

confidence: 99%

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The Effect of Tunicamycin on Interferon- Production in a Human Lymphoblastoid Cell Line (Namalwa)

Morser

1982

Journal of General Virology

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“…Purified Hu IFN-a derived from Namalwa cells induced by Sendai virus was used as a control. This IFN is known to contain at least eight subtypes of cMFN (12) and had a specific activity of 2.8 X l0 s U/mg protein.…”

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confidence: 99%

Immune human lymphocytes produce an acid-labile α-interferon

Balkwill¹,

Griffin²,

Band³

et al. 1983

The Journal of Experimental Medicine

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We have described in this paper a novel human interferon (IFN) with antigenic and cross-species reactivity of α-IFN and physicochemical properties of γ-IFN. This IFN is produced by normal peripheral blood mononuclear cells during an immune response but has also been associated with autoimmune disease (10). The system described here will be useful in elucidating the biological significance and cell of origin of this IFN.

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Analysis and Purification of Human Lymphoblastoid (Namalwa) Interferon Using a Monoclonal Antibody

Cited by 61 publications

References 22 publications

Janus Kinase-dependent Activation of Insulin Receptor Substrate 1 in Response to Interleukin-4, Oncostatin M, and the Interferons

Janus Kinase-dependent Activation of Insulin Receptor Substrate 1 in Response to Interleukin-4, Oncostatin M, and the Interferons

The Effect of Tunicamycin on Interferon- Production in a Human Lymphoblastoid Cell Line (Namalwa)

Immune human lymphocytes produce an acid-labile α-interferon

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