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

Annika Pettersson

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

Annika Pettersson

Department of Experimental Medical Science

Research output: Thesis › Doctoral Thesis (compilation)

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 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.

Original language	English
Qualification	Doctor
Awarding Institution
Supervisors/Advisors	Owman, Christer, Supervisor
Award date	2005 Apr 22
Publisher	Department of Physiological Sciences, Lund University
ISBN (Print)	91-85439-19-3
Publication status	Published - 2005

Bibliographical note

Defence details

Date: 2005-04-22
Time: 09:00
Place: Segerfalksalen, WNC, Lund

External reviewer(s)

Name: Claesson, Hans-Erik
Title: Professor
Affiliation: Karolinska Institutet, Stockholm

---

<div class="article_info">Annika Pettersson, Åke Boketoft, Alan Sabirsh, Niclas E. Nilsson, Knut Kotarsky, Björn Olde and Christer Owman. 2000. First-Generation Monoclonal Antibodies Identifying the Human Leukotriene B4 Receptor-1 Biochemical and Biophysical Research Communications, vol 279 pp 520-525. Academic Press</div>
<div class="article_info">Annika Pettersson, Johan Richter and Christer Owman. 2003. Flow cytometric mapping of the leukotriene B4 receptor, BLT1, in human bone marrow and peripheral blood using specific monoclonal antibodies International Immunopharmacology, vol 3 pp 1467-1475. Elsevier</div>
<div class="article_info">Alan Sabirsh, Annika Pettersson, Åke Boketoft, Knut Kotarsky and Christer Owman. 2003. Differential inhibition of receptor activation by two mouse monoclonal antibodies specific for the human leukotriene B4 receptor, BLT1 International Immunopharmacology, vol 3 pp 1829-1839. Elsevier</div>
<div class="article_info">Annika Pettersson, Alan Sabirsh, Jesper Bristulf, Karin Kidd-Ljunggren, Bengt Ljungberg, Christer Owman and Ulf Karlsson. 2005. Pro- and anti-inflammatory substances modulate expression of the leukotriene B4 receptor, BLT1, in human monocytes Journal of leukocyte biology, vol 77 The society for Leukocyte Biology (inpress)</div>

Subject classification (UKÄ)

Pharmacology and Toxicology

Free keywords

serologi
transplantation
Immunologi
Immunology
serology
antibodies
BLT1
LTB4

Cite this

@phdthesis{6303429f2ff24b0aac86bf822b713435,

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

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 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.",

keywords = "serologi, transplantation, Immunologi, Immunology, serology, antibodies, BLT1, LTB4",

author = "Annika Pettersson",

note = "Defence details Date: 2005-04-22 Time: 09:00 Place: Segerfalksalen, WNC, Lund External reviewer(s) Name: Claesson, Hans-Erik Title: Professor Affiliation: Karolinska Institutet, Stockholm --- <div class={"}article_info{"}>Annika Pettersson, {\AA}ke Boketoft, Alan Sabirsh, Niclas E. Nilsson, Knut Kotarsky, Bj{\"o}rn Olde and Christer Owman. 2000. First-Generation Monoclonal Antibodies Identifying the Human Leukotriene B4 Receptor-1 Biochemical and Biophysical Research Communications, vol 279 pp 520-525. Academic Press</div> <div class={"}article_info{"}>Annika Pettersson, Johan Richter and Christer Owman. 2003. Flow cytometric mapping of the leukotriene B4 receptor, BLT1, in human bone marrow and peripheral blood using specific monoclonal antibodies International Immunopharmacology, vol 3 pp 1467-1475. Elsevier</div> <div class={"}article_info{"}>Alan Sabirsh, Annika Pettersson, {\AA}ke Boketoft, Knut Kotarsky and Christer Owman. 2003. Differential inhibition of receptor activation by two mouse monoclonal antibodies specific for the human leukotriene B4 receptor, BLT1 International Immunopharmacology, vol 3 pp 1829-1839. Elsevier</div> <div class={"}article_info{"}>Annika Pettersson, Alan Sabirsh, Jesper Bristulf, Karin Kidd-Ljunggren, Bengt Ljungberg, Christer Owman and Ulf Karlsson. 2005. Pro- and anti-inflammatory substances modulate expression of the leukotriene B4 receptor, BLT1, in human monocytes Journal of leukocyte biology, vol 77 The society for Leukocyte Biology (inpress)</div>",

year = "2005",

language = "English",

isbn = "91-85439-19-3",

publisher = "Department of Physiological Sciences, Lund University",

type = "Doctoral Thesis (compilation)",

}

TY - THES

T1 - Development and application of monoclonal antibodies in studies of the human leukotriene B4 receptor, BLT1

AU - Pettersson, Annika

N1 - Defence details Date: 2005-04-22 Time: 09:00 Place: Segerfalksalen, WNC, Lund External reviewer(s) Name: Claesson, Hans-Erik Title: Professor Affiliation: Karolinska Institutet, Stockholm --- <div class="article_info">Annika Pettersson, Åke Boketoft, Alan Sabirsh, Niclas E. Nilsson, Knut Kotarsky, Björn Olde and Christer Owman. 2000. First-Generation Monoclonal Antibodies Identifying the Human Leukotriene B4 Receptor-1 Biochemical and Biophysical Research Communications, vol 279 pp 520-525. Academic Press</div> <div class="article_info">Annika Pettersson, Johan Richter and Christer Owman. 2003. Flow cytometric mapping of the leukotriene B4 receptor, BLT1, in human bone marrow and peripheral blood using specific monoclonal antibodies International Immunopharmacology, vol 3 pp 1467-1475. Elsevier</div> <div class="article_info">Alan Sabirsh, Annika Pettersson, Åke Boketoft, Knut Kotarsky and Christer Owman. 2003. Differential inhibition of receptor activation by two mouse monoclonal antibodies specific for the human leukotriene B4 receptor, BLT1 International Immunopharmacology, vol 3 pp 1829-1839. Elsevier</div> <div class="article_info">Annika Pettersson, Alan Sabirsh, Jesper Bristulf, Karin Kidd-Ljunggren, Bengt Ljungberg, Christer Owman and Ulf Karlsson. 2005. Pro- and anti-inflammatory substances modulate expression of the leukotriene B4 receptor, BLT1, in human monocytes Journal of leukocyte biology, vol 77 The society for Leukocyte Biology (inpress)</div>

PY - 2005

Y1 - 2005

N2 - 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.

AB - 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.

KW - serologi

KW - transplantation

KW - Immunologi

KW - Immunology

KW - serology

KW - antibodies

KW - BLT1

KW - LTB4

M3 - Doctoral Thesis (compilation)

SN - 91-85439-19-3

PB - Department of Physiological Sciences, Lund University

ER -

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

Abstract

Bibliographical note

Subject classification (UKÄ)

Free keywords

Fingerprint

Cite this