Analysis of apoptosis in peripheral blood and synovial tissue very early after initiation of infliximab treatment in rheumatoid arthritis patients.
OBJECTIVE
Infliximab treatment results in a decrease in synovial cellularity as early as 48 hours after initiation of therapy in patients with rheumatoid arthritis (RA). This study was undertaken to investigate whether infliximab induces apoptosis within the first 24 hours after infusion.


METHODS
The percentage of apoptotic cells was determined by flow cytometry in blood drawn from 21 patients directly before, 1 hour after, and 24 hours after infliximab infusion. Synovial tissue samples obtained before, 1 hour after (n = 5), or 24 hours after (n = 5) initiation of therapy were subjected to immunohistochemistry to detect active caspase 3 and to TUNEL assay and electron microscopy to detect apoptosis. In addition, plasma levels of nucleosomes (generated during apoptosis) and C4b/c (an indicator of complement activation) were measured.


RESULTS
There were no signs of apoptosis induction in peripheral blood monocytes or lymphocytes after infliximab treatment. Circulating lymphocyte counts were increased within 1 hour after infusion (P < 0.05). There was no definite evidence of apoptosis induction in the synovium, except in 1 patient 24 hours after the infliximab infusion. Consistent with these results, there was no increase in nucleosome levels nor were there signs of complement activation.


CONCLUSIONS
Our findings indicate that the rapid decrease in synovial cellularity observed after initiation of anti-tumor necrosis factor antibody therapy cannot be explained by apoptosis induction at the site of inflammation. It is tempting to speculate that the striking effects on synovial inflammation may be explained by other mechanisms, such as decreased migration toward the synovial compartment and reduced retention in the inflamed synovium.
Wijbrandts
Carla A
CA
Academic Medical Center/University of Amsterdam, Amsterdam, The Netherlands.
Remans
Philip H
PH
Klarenbeek
Paul L
PL
Wouters
Diana
D
van den Bergh Weerman
Marius A
MA
Smeets
Tom J
TJ
Vervoordeldonk
Margriet J
MJ
Baeten
Dominique
D
Tak
Paul P
PP
D000911:Antibodies, Monoclonal / Q000494:pharmacology* / Q000627:therapeutic use; D018501:Antirheumatic Agents / Q000494:pharmacology* / Q000627:therapeutic use; D017209:Apoptosis / Q000187:drug effects*; D001172:Arthritis, Rheumatoid / Q000188:drug therapy*; D001773:Blood Cells / Q000166:cytology / Q000187:drug effects*; D053148:Caspase 3 / Q000032:analysis; D015935:Complement C4b; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D000069285:Infliximab; D018655:Lymphocyte Count; D008214:Lymphocytes / Q000166:cytology; D008297:Male; D008854:Microscopy, Electron; D008875:Middle Aged; D009000:Monocytes / Q000166:cytology; D009707:Nucleosomes / Q000187:drug effects; D013583:Synovial Membrane / Q000166:cytology / Q000187:drug effects*
D000911:Antibodies, Monoclonal; D018501:Antirheumatic Agents; D009707:Nucleosomes; D015935:Complement C4b; D000069285:Infliximab; D053148:Caspase 3