Study
αCT Project N°1-B
February 2017

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

February 2017

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