Accessing Disease End-Points in IBD Models

High quality histopathology analysis is standard for all our IBD service models. In addition, Epistem offer a wide range of analytical services that allow Sponsors to acquire quantitative experimental data for a variety of disease-relevant end-points. Such data may help in optimising dosing regimens for therapeutic agents and can potentially identify earlier end-points in colitis development that can be used to index efficacy and reduce study time.

ibd-fig-1

Figure 1: Endpoints in IBD studies.

Profiling changes in peripheral blood leukocytes

Whole blood can be obtained at regular intervals during chronic colitis studies and can be useful for monitoring disease/treatment-related changes in number and proportion of the different leukocyte sub-populations in the peripheral circulation.

ibd-fig-2

Figure 2: Typical changes in peripheral blood leukocyte sub-populations in Prkdcscid mice (NEUT, neutrophils; LYMPH, lymphocytes; MONO, monocytes), at times following adoptive transfer of CD4+CD62L+ T cells (mean ± SD are shown).

FACS analysis of leukocytes isolated from the intestine, spleen and lymph nodes

FACS analysis of the differential expression of surface markers and both intracellular and secreted proteins can be used to define the phenotypes of isolated cell populations, which can be useful for target validation, optimisation of dosing regimens for therapeutic agents and monitoring disease development and the efficacy of experimental therapeutics.

ibd-fig-3

Figure 3: The density plots in panel A demonstrate the accumulation of CD45+CD3+ lymphocytes within the large bowel lamina propria of Prkdcscid mice, at various times following the adoptive transfer of CD4+CD62L+ T cells. The histogram plots (panel B) show that the vast majority of these lymphocytes are CD4+ T cells.

ibd-fig-4

Figure 4: The cell population shown in panel A, has distinct CD11b+ and CD4+ cell populations, which can be further differentiated on the basis of intracellular cytokine staining, with the CD11b+ cells demonstrating specific expression of cytokine “a”.

Multiplex cytokine analysis in intestinal lysates and serum/plasma

Serum/plasma, whole-tissue lysates and culture medium from ex vivo cultures can be used for multiplex cytokine analysis. In IBD models, levels of cytokine measureable in tissue lysates may be up to x10 higher than those observed in serum/plasma samples. The up-regulation of specific cytokines is associated with colitis development in the models, and treatment effects with therapeutic agents can be demonstrated.

ibd-fig-5

Figure 5: Up-regulation of TNF-α in CD4+CD62L+ T cell-recipient Prkdcscid mice is inhibited by anti‑IL‑12/IL‑23 p40 IgG.

Gene expression services

Whole tissues and isolated cells can also be processed using Epistem’s proprietary methods for analysis of gene expression. Epistem offers a wide range of services for this purpose including microarray analysis, NGS and qPCR; a full data analysis package can also be included. Laser Capture Microdissection is available for more specific analysis of whole tissues, which can allow the user to contrast the expression of inflammatory markers in adjacent “normal” and “diseased” areas.

ibd-fig-6

Figure 6: Principal component analysis of data from four experimental groups (shown by specific coloured icons) in a DSS-induced colitis study, demonstrating overlapping and non-overlapping patterns of gene expression.

For more information click here to view our website or download our recent webinar.

Get in touch

Find out how Epistem can assist with your research…

* Required Fields

Speak to one of our scientists

About Epistem

Epistem's contract research service is committed to providing reliable, innovative and transferable pre-clinical models and services to support decision making throughout the drug discovery and development pipeline.

Tel: +44 (0)161 850 7600  Email: info@epistem.co.uk