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Development and application of monoclonal antibodies in studies of the human leukotriene B4 receptor, BLT1

Pettersson, Annika LU (2005)
Abstract
Infection and inflammation are characterized by the release of chemotactic substances that activate cells of the immune system. One of these important mediators is LTB4, an arachidonic acid derivate that attracts e.g. monocytes and granulocytes to inflammatory sites and induces phagocytosis and degranulation. LTB4 binds to BLT1 which is a 7-transmembrane receptor that belongs to the superfamily of G protein-coupled receptors (GPCRs). BLT1 and LTB4 are not only important in physiological processes but also in several pathological conditions such as psoriasis, arthritis and atherosclerosis.



Monoclonal antibodies recognizing BLT1 were developed using the hybridoma technique and they were extensively characterized with... (More)
Infection and inflammation are characterized by the release of chemotactic substances that activate cells of the immune system. One of these important mediators is LTB4, an arachidonic acid derivate that attracts e.g. monocytes and granulocytes to inflammatory sites and induces phagocytosis and degranulation. LTB4 binds to BLT1 which is a 7-transmembrane receptor that belongs to the superfamily of G protein-coupled receptors (GPCRs). BLT1 and LTB4 are not only important in physiological processes but also in several pathological conditions such as psoriasis, arthritis and atherosclerosis.



Monoclonal antibodies recognizing BLT1 were developed using the hybridoma technique and they were extensively characterized with regard to specificity and applicability. The resulting two antibodies were then applied in a flow cytometric mapping of human peripheral blood and bone marrow where the highest expression of BLT1 was seen in myeloid cells.



One of the antibodies was found to inhibit LTB4 binding to BLT1 while both antibodies could inhibit receptor activation and chemotaxis towards LTB4. These findings demonstrated that one antibody was a competitive antagonist, and the other a non-competitive antagonist.



Myeloid expression of BLT1 was further investigated in monocytes using real-time PCR and flow cytomtery. Pro- and anti-inflammatory mediators were shown to modulate both mRNA and protein levels. A closer analysis of IFN-g-mediated effects was conducted and this mediator down-modulated BLT1 mRNA in a time and concentration dependent manner as well as inhibited LTB4 mediated chemotaxis. (Less)
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author
supervisor
opponent
  • Professor Claesson, Hans-Erik, Karolinska Institutet, Stockholm
organization
publishing date
type
Thesis
publication status
published
subject
keywords
serologi, transplantation, Immunologi, Immunology, serology, antibodies, BLT1, LTB4
pages
110 pages
publisher
Department of Physiological Sciences, Lund University
defense location
Segerfalksalen, WNC, Lund
defense date
2005-04-22 09:00
ISSN
1652-8220
ISBN
91-85439-19-3
language
English
LU publication?
yes
id
6303429f-2ff2-4b0a-ac86-bf822b713435 (old id 544584)
date added to LUP
2007-10-13 13:05:34
date last changed
2016-09-19 08:44:55
@phdthesis{6303429f-2ff2-4b0a-ac86-bf822b713435,
  abstract     = {Infection and inflammation are characterized by the release of chemotactic substances that activate cells of the immune system. One of these important mediators is LTB4, an arachidonic acid derivate that attracts e.g. monocytes and granulocytes to inflammatory sites and induces phagocytosis and degranulation. LTB4 binds to BLT1 which is a 7-transmembrane receptor that belongs to the superfamily of G protein-coupled receptors (GPCRs). BLT1 and LTB4 are not only important in physiological processes but also in several pathological conditions such as psoriasis, arthritis and atherosclerosis.<br/><br>
<br/><br>
Monoclonal antibodies recognizing BLT1 were developed using the hybridoma technique and they were extensively characterized with regard to specificity and applicability. The resulting two antibodies were then applied in a flow cytometric mapping of human peripheral blood and bone marrow where the highest expression of BLT1 was seen in myeloid cells.<br/><br>
<br/><br>
One of the antibodies was found to inhibit LTB4 binding to BLT1 while both antibodies could inhibit receptor activation and chemotaxis towards LTB4. These findings demonstrated that one antibody was a competitive antagonist, and the other a non-competitive antagonist.<br/><br>
<br/><br>
Myeloid expression of BLT1 was further investigated in monocytes using real-time PCR and flow cytomtery. Pro- and anti-inflammatory mediators were shown to modulate both mRNA and protein levels. A closer analysis of IFN-g-mediated effects was conducted and this mediator down-modulated BLT1 mRNA in a time and concentration dependent manner as well as inhibited LTB4 mediated chemotaxis.},
  author       = {Pettersson, Annika},
  isbn         = {91-85439-19-3},
  issn         = {1652-8220},
  keyword      = {serologi,transplantation,Immunologi,Immunology,serology,antibodies,BLT1,LTB4},
  language     = {eng},
  pages        = {110},
  publisher    = {Department of Physiological Sciences, Lund University},
  school       = {Lund University},
  title        = {Development and application of monoclonal antibodies in studies of the human leukotriene B4 receptor, BLT1},
  year         = {2005},
}