Therapeutic potency of IL2-caspase 3 targeted treatment in a murine experimental model of inflammatory bowel disease.
BACKGROUND
Inflammatory bowel disease (IBD) comprises primarily the two disorders - ulcerative colitis and Crohn's disease - that involve deregulated T cell responses. The ever-increasing incidence rate of Crohn's disease and ulcerative colitis during recent decades, combined with the limited efficacy and potential adverse effects of current treatments, explain the real need for seeking more specific and selective methods for treating these diseases.


OBJECTIVE
To investigate the ability of interleukin 2 (IL2)-caspase 3 chimeric protein, designed to target activated T lymphocytes that express the high-affinity IL2 receptor, to ameliorate the clinical symptoms of acute murine experimental colitis, using a mouse model of dextran sodium sulfate (DSS)-induced colitis.


METHODS
Mice with DSS-induced colitis were treated with IL2-caspase 3 for 7 days and disease severity was assessed in parallel to control, non-treated mice, receiving only daily injections of phosphate-buffered saline. IL2-caspase 3 was tested both for its ability to prevent the development of colitis, and for its therapeutic potential to cure on-going, active acute disease. In addition, colon tissue samples were used for myeloperoxidase assays and RNA isolation followed by polymerase chain reaction to determine mRNA expression levels of specific genes.


RESULTS
Treatment with IL2-caspase 3 dose-dependently ameliorated the disease activity index (DAI) of mice colitis. We achieved up to 78% improvement in DAI with intravenous injections of 15 microg/mouse/day. Furthermore, IL2-caspase 3 decreased neutrophil and macrophage infiltration to the inflamed tissue by up to 57%. IL2-caspase 3 was proven as a therapeutic reagent in another model, where treatment begins only after disease onset. Here we demonstrated a 70% decrease in DAI when compared to non-treated sick mice. A reduction in mRNA expression levels of both IL1 beta and tumour necrosis factor alpha was found in lysates of total colon tissue of treated mice, as compared to sick, untreated mice. We also found that expression levels of Bcl2 were significantly decreased after treatment, while Bax was upregulated in comparison to non-treated mice. Moreover, the Bcl2/Bax ratio, which is elevated in both experimental colitis and in human Crohn's disease, decreased dramatically after treatment.


CONCLUSIONS
IL2-caspase 3 chimeric protein may provide a novel approach to the therapy of human IBD, and a possible suggested treatment for other pathological conditions that involve uncontrolled expansion of activated T cells.
Shteingart
S
S
Department of Cellular Biochemistry and Human Genetics, Hebrew University, Faculty of Medicine, Jerusalem 91120, Israel.
Rapoport
M
M
Grodzovski
I
I
Sabag
O
O
Lichtenstein
M
M
Eavri
R
R
Lorberboum-Galski
H
H
D000208:Acute Disease; D000818:Animals; D053148:Caspase 3 / Q000235:genetics* / Q000378:metabolism; D003092:Colitis / Q000188:drug therapy* / Q000201:enzymology / Q000276:immunology; D003106:Colon / Q000187:drug effects / Q000276:immunology; D016264:Dextran Sulfate; D004305:Dose-Response Relationship, Drug; D015870:Gene Expression / Q000187:drug effects; D019254:Genes, bcl-2; D005818:Genetic Engineering; D007376:Interleukin-2 / Q000235:genetics* / Q000378:metabolism; D008213:Lymphocyte Activation; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D023421:Models, Animal; D009504:Neutrophils / Q000276:immunology; D012333:RNA, Messenger / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000627:therapeutic use*; D013601:T-Lymphocytes / Q000276:immunology
D007376:Interleukin-2; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D016264:Dextran Sulfate; D053148:Caspase 3