Campbell, BJ

Duckworth, CA

Elramli, AH

Pritchard, DM

AH Elramli

CA Duckworth

BJ Campbell

DM Pritchard

Crossref

PTU-047 Anti-CD3 Antibody Induces T-Cell Mediated Apoptosis and Shedding of Murine Small Intestinal Epithelial Cells

University of Liverpool Repository

BSG 2016 - Abstract SubmissionInflammatory Bowel DiseaseBSG16-ABS-1253ANTI-CD3 ANTIBODY INDUCES T-CELL MEDIATED APOPTOSIS AND SHEDDING OF MURINE SMALLINTESTINAL EPITHELIAL CELLSA. H. Elramli* 1, C. A. Duckworth1, B. J. Campbell1, D. M. Pritchard11Gastroenterology, Institute of Translational medicine, Liverpool, United KingdomDoes this abstract contain original data?: YesWill this abstract be published/presented prior to June 2016?: NoThis abstract is: A basic science submissionDoes your Endoscopy abstract include a video?: NoPreferred presentation type: Oral or PosterIntroduction: Anti-CD3 antibody binds to the CD3/TCR complex on the wall of T lymphocytes and results in theiractivation. This leads to the release of various cytokines including tumour necrosis factor (TNF). TNF has previously beenshown to bind to the TNFR1 receptor on the basolateral wall of small intestinal epithelial cells (IECs), triggering theirapoptosis and shedding (Williams JM et al, DMM, 2013). We aimed to determine the time course and dose response ofanti-CD3 antibody-induced apoptosis in murine small IECs and investigate whether this is regulated by the expression ofmembers of the NFκB family of proteins.Methods: Groups of 3 wild-type female C57BL/6J mice were injected intraperitoneally (i.p.) with 1mg/kg anti-CD3antibody and were euthanased at different time-points from 1-6 hours. Groups of 3 wild-type female C57BL/6J weresubsequently injected i.p. with 0.5-4 mg/kg anti-CD3 antibody and were killed after 1.5 hours. The responses of 6 NFκB1-/-, 6 NFκB2-/- and 6 c-Rel-/- female mice were compared with 6 C57BL/6 mice 1.5 hours after i.p. administration of 2mg/kganti-CD3 antibody. After euthanasia, the small intestine was dissected, fixed in formalin and paraffin embedded toproduce histological slides. Immunohistochemistry was performed using a rabbit anti-mouse active caspase 3 primaryantibody. Positively stained cells reflecting the percentage of shedding and apoptotic cells were scored on 20 villi permouse on a cell positional basis. Data are expressed as mean ± SEM. Statistical analysis was performed using one-wayANOVA (with Tukey post-hoc test for multiple comparisons). p<0.05 was considered significant.Results: 1mg/kg anti-CD3 antibody caused a significant increase in the percentage of IECs undergoing apoptosis andshedding from 1±0.1% in untreated mice to a peak of 2.3±0.1% at 1.5 hours then started to decline to reach 1.3±0.1% at6 hours. The percentage of IECs undergoing apoptosis and shedding increased with increasing doses of anti-CD3antibody with a maximum of 5.6±0.2% observed 1.5 hours after administration of 4mg/kg anti-CD3 antibody. AlthoughNFκB1-/- mice (5.1±0.3%) and c-Rel-/- mice (4.8±0.6%) showed more IEC apoptosis and shedding than wild-type(3.6±0.2%) these differences were not significant. However NFκB2-/- mice were much more resistant (0.8±0.1%)(p<0.001). In all cases the effects of anti-CD3 antibody were most pronounced in the apical portion of small intestinal villi.Conclusion: Systemic administration of anti-CD3 antibody induces apoptosis and shedding of murine IECs. The timecourse and cell positional distribution of apoptotic cells are very similar to those observed following administration ofeither lipopolysaccharide or TNF. The responses to anti-CD3 antibody are significantly affected by the expression ofNFκB2.Disclosure of Interest: None Declared

ANTI-CD3 ANTIBODY INDUCES T-CELL MEDIATED APOPTOSIS AND SHEDDING OF MURINE SMALL INTESTINAL EPITHELIAL CELLS

ANTI-CD3 ANTIBODY INDUCES T-CELL MEDIATED APOPTOSIS AND SHEDDING OF MURINE SMALL INTESTINAL EPITHELIAL CELLS

Abstract

Similar works

Full text

Available Versions

Crossref

University of Liverpool Repository