Skip to main content

01.12.2017 | Research article | Ausgabe 1/2017 Open Access

Arthritis Research & Therapy 1/2017

Multiplex serum protein analysis reveals potential mechanisms and markers of response to hyperimmune caprine serum in systemic sclerosis

Arthritis Research & Therapy > Ausgabe 1/2017
Niamh Quillinan, Kristina E. N. Clark, Bryan Youl, Jeffrey Vernes, Deirdre McIntosh, Syed Haq, Christopher P. Denton
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1186/​s13075-017-1252-x) contains supplementary material, which is available to authorized users.



Hyperimmune caprine serum (HICS) is a novel biological therapy with potential benefit for skin in established diffuse cutaneous systemic sclerosis. Here we report multiplex protein analysis of blood samples from a placebo-controlled phase II clinical trial and explore mechanisms of action and markers of response.


Patients were treated with HICS (n = 10) or placebo (n = 10) over 26 weeks, with follow-up open-label treatment to 52 weeks in 14 patients. Serum or plasma samples at baseline, 26 and 52 weeks were analysed using multiplex or individual immunoassays for 41 proteins. Patterns of change were analysed by clustering using Netwalker 1.0, Pearson coefficient and significance analysis of microarrays (SAM) correction.


Cluster analysis, SAM multiplex testing and paired comparison of individual analytes identified proteins that were upregulated or downregulated during treatment with HICS. There was upregulation of the hypothalamo-pituitary-adrenal axis after HICS treatment evidenced by increases in α-MSH and ACTH in cases treated with HICS. Interestingly, significant increase in PIIINP was associated with HICS treatment and improved MRSS suggesting that this may be a marker of extracellular matrix turnover. Other relevant factors reduced in HICS-treated patients compared with controls, although not reaching statistical significance included COMP, CCL2, IL6, TIMP2, Fractalkine and TGFβ1 levels.


Our results suggest mechanisms of action for HICS, including upregulation of α-MSH, that has been shown to be anti-fibrotic in preclinical models, and possible markers to be included in future trials targeting skin in diffuse cutaneous systemic sclerosis.

Trial registration

Eudract, No. 2007-003122-24., No. NCT00769028. Registered 7 October 2008.
Additional file 1: Figure S1. Subject progression in the clinical trials with serum sample time points. Schematic indicates how 20 subjects were randomly allocated to active treatment with hyperimmune caprine serum (HICS) or placebo. One patient withdrew from the HICS arm and was not available for follow-up. Other subjects were all followed to 52 weeks and at 26 weeks were offered open-label compassionate HICS. The study blind for 0–26 week treatment was maintained until after 52-week assessment. Serum and plasma samples were available for weeks 0, 26 and 52. Additional screening blood samples (pre-randomisation) were available for quality control purposes. (TIF 132 kb)
Additional file 2: Figure S2. Unsupervised hierarchical cluster analysis for multiplex serum proteins at baseline and 26 weeks for the extended dataset of 26 weeks treatment with HICS. Unsupervised cluster analysis was undertaken to identify any subgroups within the extended dataset of the study cohort at baseline, the end of placebo treatment period, or at 52 weeks based upon the serum levels of multiple protein analytes as described in text. This provided a larger sample size by including 17 subjects treated with HICS over 26 weeks and 13 subjects with 26 weeks of observation on placebo or no active treatment. The same analysis was repeated for serum samples after 26 weeks of treatment with HICS or placebo. The randomly assigned treatment allocation is shown for each subject. Data for baseline samples are shown in panel A together with treatment allocation. After 26 weeks of treatment there were clear changes in the patterns of protein analytes that were spread between the two treatment arms as shown in panel B. (TIF 1373 kb)
Additional file 3: Figure S3. Unsupervised cluster analysis for change in serum proteins from baseline to 26 weeks comparing treatment with hyperimmune caprine serum (HICS) with placebo over 26 weeks and in extended dataset at 52 weeks. (A) Unsupervised cluster analysis of change in protein level during 26-week treatment phase of placebo-controlled trial reveals patterns of change that are reflected in the supervised analysis shown in Fig.  3. Thus, subjects receiving placebo show generally less treatment effect and those treated with HICS show the patterns consistent with the summary changes shown above. (B) Unsupervised cluster analysis is also performed for the extended 52-week dataset that includes subjects moving from placebo to active treatment ( n = 7) in the second 26 weeks and three cases that have no active treatment and were previously on placebo. This complements the presentation of data for baseline and 26 weeks presented in Additional file 1: Figure S1 and shows close congruity for the two 26-week unsupervised heat maps in the extended dataset. (TIF 1444 kb)
Über diesen Artikel

Weitere Artikel der Ausgabe 1/2017

Arthritis Research & Therapy 1/2017 Zur Ausgabe

Neu im Fachgebiet Innere Medizin

Meistgelesene Bücher aus der Inneren Medizin

2017 | Buch

Rheumatologie aus der Praxis

Entzündliche Gelenkerkrankungen – mit Fallbeispielen

Dieses Fachbuch macht mit den wichtigsten chronisch entzündlichen Gelenk- und Wirbelsäulenerkrankungen vertraut. Anhand von über 40 instruktiven Fallbeispielen werden anschaulich diagnostisches Vorgehen, therapeutisches Ansprechen und der Verlauf …

Rudolf Puchner

2016 | Buch

Ambulant erworbene Pneumonie

Was, wann, warum – Dieses Buch bietet differenzierte Diagnostik und Therapie der ambulant erworbenen Pneumonie zur sofortigen sicheren Anwendung. Entsprechend der neuesten Studien und Leitlinien aller wichtigen Fachgesellschaften.

Santiago Ewig

Mail Icon II Newsletter

Bestellen Sie unseren kostenlosen Newsletter Update Innere Medizin und bleiben Sie gut informiert – ganz bequem per eMail.

© Springer Medizin