A monoclonal rat anti-mouse EMAP II antibody that functionally neutralizes pro- and mature-EMAP II in vitro.
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.
Rajashekhar
Gangaraju
G
Department of Cellular and Integrative Physiology, Indiana Center for Vascular Biology & Medicine, IU School of Medicine, Indianapolis, IN 46202, USA.
Mitnacht-Kraus
Rita
R
Ispe
Ute
U
Garrison
Jana
J
Hou
Yonghao
Y
Taylor
Brian
B
Petrache
Irina
I
Vestweber
Dietmar
D
Clauss
Matthias
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000276:immunology / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000276:immunology; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects / Q000276:immunology; D016207:Cytokines / Q000276:immunology*; D004730:Endothelium, Vascular / Q000276:immunology; D006801:Humans; D051379:Mice; D009000:Monocytes / Q000276:immunology; D009363:Neoplasm Proteins / Q000276:immunology*; D011498:Protein Precursors / Q000276:immunology*; D016601:RNA-Binding Proteins / Q000276:immunology*; D051381:Rats; D011917:Rats, Inbred Lew; D023041:Xenograft Model Antitumor Assays / Q000379:methods
D000911:Antibodies, Monoclonal; D000970:Antineoplastic Agents; D016207:Cytokines; D009363:Neoplasm Proteins; D011498:Protein Precursors; D016601:RNA-Binding Proteins; C077219:small inducible cytokine subfamily E, member 1
J Biol Chem. 1994 Oct 7;269(40):25106-19
7929199
Am J Pathol. 2003 Jan;162(1):93-103
12507893
J Biol Chem. 2001 Jun 29;276(26):23769-76
11306575
Proc Natl Acad Sci U S A. 1998 Oct 13;95(21):12322-7
9770485
J Biol Chem. 2001 Jun 22;276(25):23028-33
11292833
J Biol Chem. 1997 Dec 19;272(51):32573-9
9405472
J Clin Invest. 2001 Jan;107(1):53-63
11134180
J Immunol. 2006 Jan 1;176(1):256-64
16365417
Trends Immunol. 2004 Apr;25(4):201-9
15039047
J Biol Chem. 1999 Aug 13;274(33):23155-9
10438485
Semin Thromb Hemost. 2000;26(5):561-9
11129413
J Biol Chem. 1992 Oct 5;267(28):20239-47
1400342
Exp Hematol. 2006 Aug;34(8):1125-32
16863920
Science. 1999 Apr 2;284(5411):147-51
10102815
Microvasc Res. 2000 Jul;60(1):70-80
10873516
Lab Invest. 2009 Jan;89(1):38-46
19002109
FASEB J. 1994 Oct;8(13):1055-60
7926371
Exp Cell Res. 2006 Jul 15;312(12):2231-7
16674941
Nat Med. 2005 May;11(5):491-8
15852018
Cancer Res. 2003 Oct 1;63(19):6405-12
14559830
Apoptosis. 2000 Apr;5(2):141-51
11232242
Proc Natl Acad Sci U S A. 2006 Oct 3;103(40):14913-8
17001013
Inflamm Res. 2007 May;56(5):175-81
17393072
FEBS Lett. 2000 Jan 21;466(1):143-7
10648830
Cancer Res. 2000 Jun 1;60(11):2850-7
10850427
Lab Invest. 2000 Oct;80(10):1533-40
11045570
J Exp Med. 1999 Aug 2;190(3):341-54
10430623
Am J Physiol Lung Cell Mol Physiol. 2002 Jun;282(6):L1239-44
12003779