Abrogation of viral interleukin-6 (vIL-6)-induced signaling by intracellular retention and neutralization of vIL-6 with an anti-vIL-6 single-chain antibody selected by phage display

Marina Kovaleva, Ingo Bussmeyer, Björn Rabe, Joachim Grötzinger, Enge Sudarman, Jutta Eichler, Udo Conrad, Stefan Rose-John, Jürgen Scheller, Marina Kovaleva

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Human herpesvirus 8 (HHV-8) encodes several putative oncogenes, which are homologues to cellular host genes known to function in cell cycle regulation, control of apoptosis, and cytokine signaling. Viral interleukin (vIL-6) is believed to play an important role in the pathogenesis of Kaposi's sarcoma as well as primary effusion lymphoma and multicentric Castleman's disease. Therefore, vIL-6 is a promising target for novel therapies directed against HHV-8-associated diseases. By phage display screening of human synthetic antibody libraries, we have selected a specific recombinant antibody, called monoclonal anti-vIL-6 (MAV), binding to vIL-6. The epitope recognized by MAV was localized on the top of the D helix of the vIL-6 protein, which is a part of receptor binding site III. Consequently, MAV specifically inhibits vIL-6-mediated growth of the primary effusion lymphoma-derived cell line BCBL-1 and blocks STAT3 phosphorylation in the human hepatoma cell line HepG2. Since it was previously found that vIL-6 can also induce signals from within the cell, presumably within the endoplasmic reticulum, we fused the recombinant antibody MAV with the endoplasmic retention sequence KDEL (MAV-KDEL). As a result, COS-7 cells expressing MAV-KDEL and synthesizing vIL-6 ceased to secrete the cytokine. Moreover, we observed that vIL-6 that was bound to MAV-KDEL and retained in the endoplasmic reticulum did not induce STAT3 phosphorylation in HepG2 cells. We conclude that the activity of the intracellularly retained vIL-6 protein is neutralized by MAV-KDEL. Our results might represent a novel therapeutic strategy to neutralize virally encoded growth factors or oncogenes.
Original languageEnglish
Pages (from-to)8510-20
Number of pages11
JournalJournal of Virology
Volume80
Issue number17
DOIs
Publication statusPublished - Sep 2006

Fingerprint

Single-Chain Antibodies
interleukin-6
neutralization
bacteriophages
Bacteriophages
Interleukin-6
antibodies
Human herpesvirus 8
Primary Effusion Lymphoma
recombinant antibodies
Human Herpesvirus 8
oncogenes
lymphoma
Oncogenes
Endoplasmic Reticulum
endoplasmic reticulum
lysyl-aspartyl-glutamyl-leucine
phosphorylation
cytokines
Monoclonal Antibodies

Keywords

  • Animals
  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Antibody Specificity
  • COS Cells
  • Cell Line
  • Cercopithecus aethiops
  • Endoplasmic Reticulum
  • Escherichia coli
  • Humans
  • Immunoglobulin Variable Region
  • Interleukin-6
  • Neutralization Tests
  • Peptide Library
  • Recombinant Proteins
  • Signal Transduction
  • Viral Proteins

Cite this

Abrogation of viral interleukin-6 (vIL-6)-induced signaling by intracellular retention and neutralization of vIL-6 with an anti-vIL-6 single-chain antibody selected by phage display. / Kovaleva, Marina; Bussmeyer, Ingo; Rabe, Björn; Grötzinger, Joachim; Sudarman, Enge; Eichler, Jutta; Conrad, Udo; Rose-John, Stefan; Scheller, Jürgen; Kovaleva, Marina.

In: Journal of Virology, Vol. 80, No. 17, 09.2006, p. 8510-20.

Research output: Contribution to journalArticle

Kovaleva, M, Bussmeyer, I, Rabe, B, Grötzinger, J, Sudarman, E, Eichler, J, Conrad, U, Rose-John, S, Scheller, J & Kovaleva, M 2006, 'Abrogation of viral interleukin-6 (vIL-6)-induced signaling by intracellular retention and neutralization of vIL-6 with an anti-vIL-6 single-chain antibody selected by phage display', Journal of Virology, vol. 80, no. 17, pp. 8510-20. https://doi.org/10.1128/JVI.00420-06
Kovaleva, Marina ; Bussmeyer, Ingo ; Rabe, Björn ; Grötzinger, Joachim ; Sudarman, Enge ; Eichler, Jutta ; Conrad, Udo ; Rose-John, Stefan ; Scheller, Jürgen ; Kovaleva, Marina. / Abrogation of viral interleukin-6 (vIL-6)-induced signaling by intracellular retention and neutralization of vIL-6 with an anti-vIL-6 single-chain antibody selected by phage display. In: Journal of Virology. 2006 ; Vol. 80, No. 17. pp. 8510-20.
@article{50f1cd67bbaa4adc89fcf67c72cf193c,
title = "Abrogation of viral interleukin-6 (vIL-6)-induced signaling by intracellular retention and neutralization of vIL-6 with an anti-vIL-6 single-chain antibody selected by phage display",
abstract = "Human herpesvirus 8 (HHV-8) encodes several putative oncogenes, which are homologues to cellular host genes known to function in cell cycle regulation, control of apoptosis, and cytokine signaling. Viral interleukin (vIL-6) is believed to play an important role in the pathogenesis of Kaposi's sarcoma as well as primary effusion lymphoma and multicentric Castleman's disease. Therefore, vIL-6 is a promising target for novel therapies directed against HHV-8-associated diseases. By phage display screening of human synthetic antibody libraries, we have selected a specific recombinant antibody, called monoclonal anti-vIL-6 (MAV), binding to vIL-6. The epitope recognized by MAV was localized on the top of the D helix of the vIL-6 protein, which is a part of receptor binding site III. Consequently, MAV specifically inhibits vIL-6-mediated growth of the primary effusion lymphoma-derived cell line BCBL-1 and blocks STAT3 phosphorylation in the human hepatoma cell line HepG2. Since it was previously found that vIL-6 can also induce signals from within the cell, presumably within the endoplasmic reticulum, we fused the recombinant antibody MAV with the endoplasmic retention sequence KDEL (MAV-KDEL). As a result, COS-7 cells expressing MAV-KDEL and synthesizing vIL-6 ceased to secrete the cytokine. Moreover, we observed that vIL-6 that was bound to MAV-KDEL and retained in the endoplasmic reticulum did not induce STAT3 phosphorylation in HepG2 cells. We conclude that the activity of the intracellularly retained vIL-6 protein is neutralized by MAV-KDEL. Our results might represent a novel therapeutic strategy to neutralize virally encoded growth factors or oncogenes.",
keywords = "Animals, Antibodies, Monoclonal, Antibodies, Viral, Antibody Specificity, COS Cells, Cell Line, Cercopithecus aethiops, Endoplasmic Reticulum, Escherichia coli, Humans, Immunoglobulin Variable Region, Interleukin-6, Neutralization Tests, Peptide Library, Recombinant Proteins, Signal Transduction, Viral Proteins",
author = "Marina Kovaleva and Ingo Bussmeyer and Bj{\"o}rn Rabe and Joachim Gr{\"o}tzinger and Enge Sudarman and Jutta Eichler and Udo Conrad and Stefan Rose-John and J{\"u}rgen Scheller and Marina Kovaleva",
year = "2006",
month = "9",
doi = "10.1128/JVI.00420-06",
language = "English",
volume = "80",
pages = "8510--20",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "17",

}

TY - JOUR

T1 - Abrogation of viral interleukin-6 (vIL-6)-induced signaling by intracellular retention and neutralization of vIL-6 with an anti-vIL-6 single-chain antibody selected by phage display

AU - Kovaleva, Marina

AU - Bussmeyer, Ingo

AU - Rabe, Björn

AU - Grötzinger, Joachim

AU - Sudarman, Enge

AU - Eichler, Jutta

AU - Conrad, Udo

AU - Rose-John, Stefan

AU - Scheller, Jürgen

AU - Kovaleva, Marina

PY - 2006/9

Y1 - 2006/9

N2 - Human herpesvirus 8 (HHV-8) encodes several putative oncogenes, which are homologues to cellular host genes known to function in cell cycle regulation, control of apoptosis, and cytokine signaling. Viral interleukin (vIL-6) is believed to play an important role in the pathogenesis of Kaposi's sarcoma as well as primary effusion lymphoma and multicentric Castleman's disease. Therefore, vIL-6 is a promising target for novel therapies directed against HHV-8-associated diseases. By phage display screening of human synthetic antibody libraries, we have selected a specific recombinant antibody, called monoclonal anti-vIL-6 (MAV), binding to vIL-6. The epitope recognized by MAV was localized on the top of the D helix of the vIL-6 protein, which is a part of receptor binding site III. Consequently, MAV specifically inhibits vIL-6-mediated growth of the primary effusion lymphoma-derived cell line BCBL-1 and blocks STAT3 phosphorylation in the human hepatoma cell line HepG2. Since it was previously found that vIL-6 can also induce signals from within the cell, presumably within the endoplasmic reticulum, we fused the recombinant antibody MAV with the endoplasmic retention sequence KDEL (MAV-KDEL). As a result, COS-7 cells expressing MAV-KDEL and synthesizing vIL-6 ceased to secrete the cytokine. Moreover, we observed that vIL-6 that was bound to MAV-KDEL and retained in the endoplasmic reticulum did not induce STAT3 phosphorylation in HepG2 cells. We conclude that the activity of the intracellularly retained vIL-6 protein is neutralized by MAV-KDEL. Our results might represent a novel therapeutic strategy to neutralize virally encoded growth factors or oncogenes.

AB - Human herpesvirus 8 (HHV-8) encodes several putative oncogenes, which are homologues to cellular host genes known to function in cell cycle regulation, control of apoptosis, and cytokine signaling. Viral interleukin (vIL-6) is believed to play an important role in the pathogenesis of Kaposi's sarcoma as well as primary effusion lymphoma and multicentric Castleman's disease. Therefore, vIL-6 is a promising target for novel therapies directed against HHV-8-associated diseases. By phage display screening of human synthetic antibody libraries, we have selected a specific recombinant antibody, called monoclonal anti-vIL-6 (MAV), binding to vIL-6. The epitope recognized by MAV was localized on the top of the D helix of the vIL-6 protein, which is a part of receptor binding site III. Consequently, MAV specifically inhibits vIL-6-mediated growth of the primary effusion lymphoma-derived cell line BCBL-1 and blocks STAT3 phosphorylation in the human hepatoma cell line HepG2. Since it was previously found that vIL-6 can also induce signals from within the cell, presumably within the endoplasmic reticulum, we fused the recombinant antibody MAV with the endoplasmic retention sequence KDEL (MAV-KDEL). As a result, COS-7 cells expressing MAV-KDEL and synthesizing vIL-6 ceased to secrete the cytokine. Moreover, we observed that vIL-6 that was bound to MAV-KDEL and retained in the endoplasmic reticulum did not induce STAT3 phosphorylation in HepG2 cells. We conclude that the activity of the intracellularly retained vIL-6 protein is neutralized by MAV-KDEL. Our results might represent a novel therapeutic strategy to neutralize virally encoded growth factors or oncogenes.

KW - Animals

KW - Antibodies, Monoclonal

KW - Antibodies, Viral

KW - Antibody Specificity

KW - COS Cells

KW - Cell Line

KW - Cercopithecus aethiops

KW - Endoplasmic Reticulum

KW - Escherichia coli

KW - Humans

KW - Immunoglobulin Variable Region

KW - Interleukin-6

KW - Neutralization Tests

KW - Peptide Library

KW - Recombinant Proteins

KW - Signal Transduction

KW - Viral Proteins

U2 - 10.1128/JVI.00420-06

DO - 10.1128/JVI.00420-06

M3 - Article

C2 - 16912301

VL - 80

SP - 8510

EP - 8520

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 17

ER -