IBD Studies
αCT Project N°1-B
Evaluation of Anti-Inflammatory Properties of AFP (ACT-101) In Vivo
In a Validated Model of Colitis Induced by TNBS (Trinitrobenzenesulfonic Acid) in Mice
Testing Facility: Intestinal Biotech Development (IBD)
Study αCT Project N°1-B
Wallace’s Score
Evaluation of Inflammation at the Macroscopic Level
| |
Mean |
sem |
p |
| TNBS + Vehicle |
6.16 |
0.23 |
|
| TNBS + AFP 200µg/day iv |
3.67 |
0.44 |
< 0.0001 |
| TNBS + mouse anti-TNFα 0.1mg ip |
3.93 |
0.34 |
< 0.0001 |
In our control treatment group the mouse anti-TNFα induces a significant decrease of inflammation compared to colitic mice receiving the vehicle, validating the model.
A significant decrease of the inflammation at the macroscopic level was observed with AFP at 200µg/mouse/day administered by iv injection.
% of Inflammation Score Improvement:
- 36% with Anti-TNFα
- 41% with AFP 200µg/day by IV administration (similar effect to anti-TNFα)
Ameho’s Score
Evaluation of Inflammation at the Histological Level
| |
Mean |
sem |
p |
| TNBS + Vehicle |
5.26 |
0.21 |
|
| TNBS + AFP 200µg/day iv |
2.94 |
0.46 |
0.00006 |
| TNBS + mouse anti-TNFα 0.1mg ip |
4.33 |
0.44 |
0.036 |
Good correlation between the macroscopic and histological scores showing a significant improvement of inflammation. Greater anti-inflammatory effect is observed with AFP compared to anti-TNFα.
Quantification of MPO
| |
Mean |
sem |
p |
| TNBS + Vehicle |
40.72 |
0.64 |
|
| TNBS + AFP 200µg/day iv |
25.42 |
4.22 |
0.00062223 |
| TNBS + mouse anti-TNFα 0.1mg ip |
29.59 |
3.11 |
1.631E-005 |
Significant decrease of MPO level in the colon, in correlation with macroscopic score and histological score indicating a reduced number of pro-inflammatory cells in the colon.
Evaluation of Gene Expression of Pro- and Anti-Inflammatory Cytokines in the Colon
Significant decrease of pro-inflammatory cytokines with AFP. Similar decrease compared to mouse anti-TNFα. Significant increase of anti-inflammatory cytokine IL10 at mRNA level.
| TNFα |
Mean |
sem |
p |
| TNBS + PBS |
22.11 |
3.77 |
|
| TNBS + AFP 200µg/day iv |
7.63 |
1.17 |
0.0009265 |
| TNBS + mouse anti-TNFα 0.1mg ip |
6.34 |
1.13 |
0.0009829 |
| IL-1b |
Mean |
sem |
p |
| TNBS + PBS |
10.97 |
3.2 |
|
| TNBS + AFP 200µg/day iv |
1.63 |
0.58 |
0.0009715 |
| TNBS + mouse anti-TNFα 0.1mg ip |
1.3 |
0.73 |
0.00382 |
| IL-6 |
Mean |
sem |
p |
| TNBS + PBS |
30.23 |
8.78 |
|
| TNBS + AFP 200µg/day iv |
6.48 |
3.3 |
0.01806 |
| TNBS + mouse anti-TNFα 0.1mg ip |
0.97 |
0.43 |
0.002747 |
| IFNγ |
Mean |
sem |
p |
| TNBS + PBS |
39.17 |
11.38 |
|
| TNBS + AFP 200µg/day iv |
7.95 |
4.18 |
0.01536 |
| TNBS + mouse anti-TNFα 0.1mg ip |
1.26 |
0.55 |
0.003788 |
| IL-10 |
Mean |
sem |
p |
| TNBS + PBS |
9.03 |
2.93 |
|
| TNBS + AFP 200µg/day iv |
68.09 |
28.87 |
0.008679 |
| TNBS + mouse anti-TNFα 0.1mg ip |
75.47 |
54.58 |
0.04176 |
