Effects of anti-β2-GPI antibodies on cytokine production in normal first-trimester trophoblast cells.
OBJECTIVE
The anti-β2-GPI antibody (aβ2-GPIAb) has been detected in recurrent fetal loss with strong pathogenic activity. The effects of aβ2-GPIAb on cytokine production and aβ2-GPIAb binding sites in first-trimester trophoblast cells were evaluated.


METHODS
First-trimester trophoblast cells were cultured in 24-well tissue culture plates with immunoglobulin G (IgG) obtained from aβ2-GPIAb-positive and aβ2-GPIAb-negative serum. Cytokines in the cultured supernatant were measured using the suspension array system and enzyme-linked immunosorbent assays. To identify potential binding sites for aβ2-GPIAb, such as toll-like receptors (TLR) 2 or TLR4, we used mouse monoclonal anti-TLR2 and/or anti-TLR4 antibodies to inhibit TLR and then measured cytokine production.


RESULTS
The production of cytokines, such as interleukin-6 and interleukin-8, increased more in response to aβ2-GPIAb-positive IgG than to aβ2-GPIAb-negative IgG in trophoblast cells. The secretion of cytokines from trophoblast cells decreased when the TLR were blocked with mouse monoclonal anti-TLR2 and anti-TLR4 antibodies.


CONCLUSIONS
We suspect that aβ2-GPIAb might increase cytokine production by binding to TLR2 or TLR4. The increased cytokine production in response to aβ2-GPIAb might play a role in the increased inflammatory response in the placenta.
Department of Obstetrics and Gynecology, Nihon University School of Medicine, Tokyo, Japan.
Hiromitsu
H
Azuma
Department of Obstetrics and Gynecology, Nihon University School of Medicine, Tokyo, Japan.
Tatsuo
T
Yamamoto
Department of Obstetrics and Gynecology, Nihon University School of Medicine, Tokyo, Japan.
Fumihisa
F
Chishima
D000818:Animals; D000906:Antibodies / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000378:metabolism; D002470:Cell Survival; D002478:Cells, Cultured; D016207:Cytokines / Q000276:immunology* / Q000378:metabolism; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000008:administration & dosage / Q000378:metabolism; D015850:Interleukin-6 / Q000276:immunology / Q000378:metabolism; D016209:Interleukin-8 / Q000276:immunology / Q000378:metabolism; D051379:Mice; D011247:Pregnancy; D011261:Pregnancy Trimester, First / Q000276:immunology*; D011485:Protein Binding; D051195:Toll-Like Receptor 2 / Q000276:immunology / Q000378:metabolism; D051197:Toll-Like Receptor 4 / Q000276:immunology / Q000378:metabolism; D014327:Trophoblasts / Q000276:immunology* / Q000378:metabolism; D053482:beta 2-Glycoprotein I / Q000276:immunology* / Q000378:metabolism
D000906:Antibodies; D000911:Antibodies, Monoclonal; D016207:Cytokines; C508600:IL6 protein, human; D007074:Immunoglobulin G; D015850:Interleukin-6; D016209:Interleukin-8; C495342:TLR2 protein, human; C495345:TLR4 protein, human; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D053482:beta 2-Glycoprotein I
anti-β2-GPI antibodies; antiphospholipid syndrome; cytokine; toll-like receptor; trophoblast