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Multiplex cytokine analyses in patients with rheumatoid arthritis require use of agents blocking heterophilic antibody activity

OLSSON, PETER LU ; Theander, E. LU ; Bergström, U. LU ; Jovinge, S. LU ; Jacobsson, LTH LU and Turesson, C. LU (2016) In Scandinavian Journal of Rheumatology
Abstract

Objectives: Heterophilic antibodies, such as rheumatoid factor (RF), are known to interfere with enzyme-linked immunosorbent assays (ELISAs). Treatment of rheumatoid arthritis (RA) with tumour necrosis factor (TNF)-α blockers is well established. The aims of this study were to develop a protocol for blocking the interaction of present heterophilic antibodies and to validate this procedure by evaluating the effect on correlations of cytokine levels to clinical response in RA patients treated with adalimumab. Method: Fourteen patients with active RA were evaluated at baseline and 3 months after starting adalimumab treatment. Cytokines were analysed with a commercial 12-plex bead ELISA. To block interference by RF, a commercial blocker... (More)

Objectives: Heterophilic antibodies, such as rheumatoid factor (RF), are known to interfere with enzyme-linked immunosorbent assays (ELISAs). Treatment of rheumatoid arthritis (RA) with tumour necrosis factor (TNF)-α blockers is well established. The aims of this study were to develop a protocol for blocking the interaction of present heterophilic antibodies and to validate this procedure by evaluating the effect on correlations of cytokine levels to clinical response in RA patients treated with adalimumab. Method: Fourteen patients with active RA were evaluated at baseline and 3 months after starting adalimumab treatment. Cytokines were analysed with a commercial 12-plex bead ELISA. To block interference by RF, a commercial blocker (HeteroBlock) was used. To determine the optimal concentration of HeteroBlock, patient sera were analysed with different concentrations of HeteroBlock. Subsequently, baseline and follow-up sera from the 14 patients were analysed and correlated with clinical outcome. Results: Measured cytokine levels were reduced in the majority of samples when adding the blocker. The optimal concentration of HeteroBlock was 1600 μg/mL of serum. Sera with high RF levels were more prone to produce false positive values, although some RF-negative sera also demonstrated evidence of interference. HeteroBlock did not interfere with the analysis. In RA patients treated with adalimumab, changes in interleukin (IL)-6 levels between baseline and follow-up correlated with changes in erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) in sera with added HeteroBlock. Conclusions: When analysing sera from patients with RA with multiplex bead ELISA, the assay should be evaluated for interference by heterophilic antibodies, and if present corrected with, for example, HeteroBlock.

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author
organization
publishing date
type
Contribution to journal
publication status
epub
subject
in
Scandinavian Journal of Rheumatology
pages
10 pages
publisher
Taylor & Francis
external identifiers
  • Scopus:84966710361
ISSN
0300-9742
DOI
10.3109/03009742.2016.1161070
language
English
LU publication?
yes
id
f8408d0b-1483-466c-a428-b904b9c30791
date added to LUP
2016-06-01 12:20:16
date last changed
2016-10-05 12:28:24
@misc{f8408d0b-1483-466c-a428-b904b9c30791,
  abstract     = {<p>Objectives: Heterophilic antibodies, such as rheumatoid factor (RF), are known to interfere with enzyme-linked immunosorbent assays (ELISAs). Treatment of rheumatoid arthritis (RA) with tumour necrosis factor (TNF)-α blockers is well established. The aims of this study were to develop a protocol for blocking the interaction of present heterophilic antibodies and to validate this procedure by evaluating the effect on correlations of cytokine levels to clinical response in RA patients treated with adalimumab. Method: Fourteen patients with active RA were evaluated at baseline and 3 months after starting adalimumab treatment. Cytokines were analysed with a commercial 12-plex bead ELISA. To block interference by RF, a commercial blocker (HeteroBlock) was used. To determine the optimal concentration of HeteroBlock, patient sera were analysed with different concentrations of HeteroBlock. Subsequently, baseline and follow-up sera from the 14 patients were analysed and correlated with clinical outcome. Results: Measured cytokine levels were reduced in the majority of samples when adding the blocker. The optimal concentration of HeteroBlock was 1600 μg/mL of serum. Sera with high RF levels were more prone to produce false positive values, although some RF-negative sera also demonstrated evidence of interference. HeteroBlock did not interfere with the analysis. In RA patients treated with adalimumab, changes in interleukin (IL)-6 levels between baseline and follow-up correlated with changes in erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) in sera with added HeteroBlock. Conclusions: When analysing sera from patients with RA with multiplex bead ELISA, the assay should be evaluated for interference by heterophilic antibodies, and if present corrected with, for example, HeteroBlock.</p>},
  author       = {OLSSON, PETER and Theander, E. and Bergström, U. and Jovinge, S. and Jacobsson, LTH and Turesson, C.},
  issn         = {0300-9742},
  language     = {eng},
  month        = {05},
  pages        = {10},
  publisher    = {ARRAY(0x9272f98)},
  series       = {Scandinavian Journal of Rheumatology},
  title        = {Multiplex cytokine analyses in patients with rheumatoid arthritis require use of agents blocking heterophilic antibody activity},
  url          = {http://dx.doi.org/10.3109/03009742.2016.1161070},
  year         = {2016},
}