A CD3-specific antibody reduces cytokine production and alters phosphoprotein profiles in intestinal tissues from patients with inflammatory bowel disease.
OBJECTIVE
T cells mediate the development of inflammation in inflammatory bowel disease (IBD). We investigated the effects of an antibody against CD3 called otelixizumab, which induces immune tolerance, in intestinal mucosa samples from patients.


METHODS
Intestinal tissues were isolated from patients undergoing routine endoscopy or from patients undergoing intestinal surgery for colon cancer or IBD; healthy surrounding tissues were collected as controls. Isolated lamina propria mononuclear cells (LPMCs) and mucosal tissue explants were incubated with otelixizumab for 24 or 48 hours. Production of inflammatory cytokines was determined by enzyme-linked immunosorbent assay. Levels of 36 cytokines and chemokines and phosphorylation of 39 receptor tyrosine kinases and signaling molecules were measured using protein arrays. Immunoblot analysis was used to analyze T-cell transcription factors.


RESULTS
Incubation of intestinal tissues or LPMCs with otelixizumab reduced production of interferon gamma, interleukin (IL)-17A, and other inflammatory cytokines and chemokines, simultaneously increasing production of IL-10. Mucosal biopsy specimens from patients with IBD retained inflammation-associated tyrosine phosphoprotein profiles ex vivo. Incubation of the inflamed tissue with otelixizumab reduced phosphorylation of these proteins to levels observed in control tissues. Otelixizumab also markedly reduced phosphorylation of proteins associated with T-cell receptor activation. Neutralization of IL-10 blocked the anti-inflammatory effects of otelixizumab.


CONCLUSIONS
We observed anti-inflammatory effects of anti-CD3 in inflamed intestinal tissues from patients with IBD. The antibody appears to down-regulate T-cell activation via IL-10.
Centre for Immunology and Infectious Disease, Barts and The London School of Medicine and Dentistry, Blizard Institute, London, England. Electronic address: a.vossenkaemper@qmul.ac.uk.
Anna
A
Vossenkämper
Institute for Functional Genomics, Computational Diagnostics Group, University of Regensburg, Regensburg, Germany.
Christian
C
Hundsrucker
GlaxoSmithKline, Pharmaceuticals R&D Facility, Stevenage, Hertfordshire, England.
Kevin
K
Page
GlaxoSmithKline, Pharmaceuticals R&D Facility, Stevenage, Hertfordshire, England.
André
A
van Maurik
Centre for Immunology and Infectious Disease, Barts and The London School of Medicine and Dentistry, Blizard Institute, London, England.
Theodore J
TJ
Sanders
Centre for Immunology and Infectious Disease, Barts and The London School of Medicine and Dentistry, Blizard Institute, London, England.
Andrew J
AJ
Stagg
Centre for Digestive Diseases, Barts and The London School of Medicine and Dentistry, Blizard Institute, London, England.
Lisa
L
Das
Centre for Immunology and Infectious Disease, Barts and The London School of Medicine and Dentistry, Blizard Institute, London, England.
Thomas T
TT
MacDonald
D000293:Adolescent; D000328:Adult; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D061067:Antibodies, Monoclonal, Humanized / Q000494:pharmacology*; D001706:Biopsy; D017252:CD3 Complex / Q000276:immunology*; D016022:Case-Control Studies; D003106:Colon / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D016207:Cytokines / Q000378:metabolism*; D005260:Female; D006801:Humans; D007108:Immune Tolerance / Q000187:drug effects; D015212:Inflammatory Bowel Diseases / Q000378:metabolism* / Q000473:pathology; D007371:Interferon-gamma / Q000378:metabolism; D016753:Interleukin-10 / Q000378:metabolism; D020381:Interleukin-17 / Q000378:metabolism; D007413:Intestinal Mucosa / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008297:Male; D008875:Middle Aged; D010750:Phosphoproteins / Q000378:metabolism*; D055815:Young Adult
D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D017252:CD3 Complex; D016207:Cytokines; D020381:Interleukin-17; D010750:Phosphoproteins; D016753:Interleukin-10; D007371:Interferon-gamma; C550701:otelixizumab
Crohn's Disease; IFN; Immune Regulation; Ulcerative Colitis