Variability in cytokine and cell adhesion molecule staining in arthroscopic synovial biopsies: quantification using color video image analysis.
OBJECTIVE
To investigate the variability in immunostaining for cytokines and cell adhesion molecules using multiple arthroscopically directed synovial biopsies from within a rheumatoid knee joint, quantitated by color video image analysis.


METHODS
Needle arthroscopic biopsies were taken from multiple sites (4-7 sites) around a knee joint in 8 patients with rheumatoid arthritis (RA). In 5 patients, immunoperoxidase staining for the cytokines tumor necrosis factor alpha (TNF-alpha), interleukin 8 (IL-8), and IL-1beta as well as the IL-1 receptor antagonist protein (IL-1ra) was performed. In 3 patients, immunoperoxidase staining for the cell adhesion molecules E-selectin (CD62E), P-selectin (CD62P), intercellular adhesion molecule 1 (ICAM-1, CD54), and platelet endothelial cell adhesion molecule (PECAM, CD31) was performed. Immunostaining was quantified using color video image analysis.


RESULTS
The overall probability of paired biopsies from the same RA knee joint being significantly different from each other due to sampling variation was at most 22% for cytokine staining (usually less than 10%). There were no significant differences between intrabiopsy and interbiopsy variability for cell adhesion molecule staining of the sublining and vessels.


CONCLUSIONS
The variability in cytokine and cell adhesion molecule staining within any single biopsy usually reflects the variability between biopsies taken from different sites in the same rheumatoid joint when the immunostaining is quantified using color video image analysis. Therefore, only a small number of synovial biopsies are required to accurately determine the cytokine and cell adhesion molecule expression in a single joint.
Youssef
P P
PP
Department of Pathology, Repatriation General Hospital, Daw Park, SA, Australia.
Triantafillou
S
S
Parker
A
A
Coleman
M
M
Roberts-Thomson
P J
PJ
Ahern
M J
MJ
Smith
M D
MD
D000368:Aged; D000369:Aged, 80 and over; D001707:Biopsy, Needle; D015815:Cell Adhesion Molecules / Q000032:analysis*; D003116:Color; D016207:Cytokines / Q000032:analysis*; D019040:E-Selectin / Q000032:analysis; D005260:Female; D006801:Humans; D019044:Image Cytometry / Q000379:methods / Q000592:standards*; D018799:Intercellular Adhesion Molecule-1 / Q000032:analysis; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1 / Q000032:analysis; D016209:Interleukin-8 / Q000032:analysis; D008297:Male; D008875:Middle Aged; D019007:P-Selectin / Q000032:analysis; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000032:analysis; D017472:Receptors, Interleukin-1 / Q000037:antagonists & inhibitors; D015203:Reproducibility of Results; D012795:Sialoglycoproteins / Q000032:analysis; D013583:Synovial Membrane / Q000737:chemistry* / Q000473:pathology*; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis
D015815:Cell Adhesion Molecules; D016207:Cytokines; D019040:E-Selectin; C507763:IL1RN protein, human; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1; D016209:Interleukin-8; D019007:P-Selectin; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D017472:Receptors, Interleukin-1; D012795:Sialoglycoproteins; D014409:Tumor Necrosis Factor-alpha; D018799:Intercellular Adhesion Molecule-1