A monoclonal rat anti-mouse EMAP II antibody that functionally neutralizes pro- and mature-EMAP II in vitro

Raja Shekhar Gangaraju, Rita Mitnacht-Kraus, Ute Ispe, Jana Garrison, Yonghao Hou, Brian Taylor, Irina Petrache, Dietmar Vestweber, Matthias Clauss

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

EMAP II is an endothelial cell and monocyte activating proinflammatory cytokine, which has been demonstrated to induce endothelial cell apoptosis. In order to analyze its role in disease models linked to inflammation and endothelial cell death, we aimed to develop a neutralizing antibody against mouse EMAP II. Therefore, we generated rat monoclonal anti-mouse EMAP II antibodies by immunization with recombinant full length, mouse pro-EMAP II protein. We could identify by ELISA, hybridoma clones from fusion with mouse myeloma SP2/0 cells which produced antibodies recognizing both full length and mature EMAP II. We further characterized one antibody, M7/1 and demonstrated its ability to detect both EMAP II forms in Western blotting and to neutralize EMAP II directed migration of human peripheral blood monocytes as well as EMAP II induced apoptosis of tumor and endothelial cells. We conclude that this antibody can be useful to both target and analyze murine disease models, in which EMAP II may be involved.

Original languageEnglish (US)
Pages (from-to)22-28
Number of pages7
JournalJournal of Immunological Methods
Volume350
Issue number1-2
DOIs
StatePublished - Oct 31 2009

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Antibodies
Endothelial Cells
Monocytes
Apoptosis
In Vitro Techniques
small inducible cytokine subfamily E, member 1
Hybridomas
Neutralizing Antibodies
Immunization
Cell Death
Clone Cells
Western Blotting
Enzyme-Linked Immunosorbent Assay
Cytokines
Inflammation
Neoplasms
Proteins

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

Cite this

A monoclonal rat anti-mouse EMAP II antibody that functionally neutralizes pro- and mature-EMAP II in vitro. / Gangaraju, Raja Shekhar; Mitnacht-Kraus, Rita; Ispe, Ute; Garrison, Jana; Hou, Yonghao; Taylor, Brian; Petrache, Irina; Vestweber, Dietmar; Clauss, Matthias.

In: Journal of Immunological Methods, Vol. 350, No. 1-2, 31.10.2009, p. 22-28.

Research output: Contribution to journalArticle

Gangaraju, RS, Mitnacht-Kraus, R, Ispe, U, Garrison, J, Hou, Y, Taylor, B, Petrache, I, Vestweber, D & Clauss, M 2009, 'A monoclonal rat anti-mouse EMAP II antibody that functionally neutralizes pro- and mature-EMAP II in vitro', Journal of Immunological Methods, vol. 350, no. 1-2, pp. 22-28. https://doi.org/10.1016/j.jim.2009.08.003
Gangaraju, Raja Shekhar ; Mitnacht-Kraus, Rita ; Ispe, Ute ; Garrison, Jana ; Hou, Yonghao ; Taylor, Brian ; Petrache, Irina ; Vestweber, Dietmar ; Clauss, Matthias. / A monoclonal rat anti-mouse EMAP II antibody that functionally neutralizes pro- and mature-EMAP II in vitro. In: Journal of Immunological Methods. 2009 ; Vol. 350, No. 1-2. pp. 22-28.
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AU - Gangaraju, Raja Shekhar

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AU - Garrison, Jana

AU - Hou, Yonghao

AU - Taylor, Brian

AU - Petrache, Irina

AU - Vestweber, Dietmar

AU - Clauss, Matthias

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AB - EMAP II is an endothelial cell and monocyte activating proinflammatory cytokine, which has been demonstrated to induce endothelial cell apoptosis. In order to analyze its role in disease models linked to inflammation and endothelial cell death, we aimed to develop a neutralizing antibody against mouse EMAP II. Therefore, we generated rat monoclonal anti-mouse EMAP II antibodies by immunization with recombinant full length, mouse pro-EMAP II protein. We could identify by ELISA, hybridoma clones from fusion with mouse myeloma SP2/0 cells which produced antibodies recognizing both full length and mature EMAP II. We further characterized one antibody, M7/1 and demonstrated its ability to detect both EMAP II forms in Western blotting and to neutralize EMAP II directed migration of human peripheral blood monocytes as well as EMAP II induced apoptosis of tumor and endothelial cells. We conclude that this antibody can be useful to both target and analyze murine disease models, in which EMAP II may be involved.

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