CHEMOREPULSION OF CELLS
The present invention provides compositions and methods of controlling the direction and/or movement of migratory cells. Specifically, the invention is directed to the identification of novel chemorepellents and unimodal fugetaxins, their agonists and antagonists which alter or affect the movement of cells involved in immune, inflammatory or cancerous phenotypes.
1 . A method of inducing negative chemotaxis in human migratory cells comprising contacting said human migratory cells with a validated chemorepellent.
2 . The method of claim 1 wherein the human migratory cells are immune cells selected from the group consisting of neutrophils, CD4+ and CD8+ T cells, B cells, monocytes or dendritic cells.
3 . The method of claim 2 wherein the validated chemorepellent is selected from the group consisting of carbohydrate binding proteins, serpins, bacterial cell wall components, heat shock proteins, natural products, Toll-like receptor ligands, viral factors, semaphorins, elastase inhibitors, antibiotics, muscle cell proteins, plant cell wall components and chemokines.
4 . The method of claim 3 wherein the immune cells are neutrophils, the validated chemorepellent is a carbohydrate binding protein and wherein said carbohydrate binding protein is galectin-1.
5 . The method of claim 3 wherein the immune cells are neutrophils, the validated chemorepellent is a serpin and wherein said serpin is antithrombin III.
6 . The method of claim 3 wherein the immune cells are neutrophils, the validated chemorepellent is a bacterial cell wall component and wherein said bacterial cell wall component is peptidoglycan from S. aureus.
7 . The method of claim 3 wherein the immune cells are neutrophils, the validated chemorepellent is a heat shock protein and wherein said heat shock protein is heat shock protein 25.
8 . The method of claim 3 wherein the immune cells are neutrophils, the validated chemorepellent is a natural product and wherein said natural product is reservatrol.
9 . The method of claim 3 wherein the immune cells are neutrophils, the validated chemorepellent is a Toll-like receptor ligands and wherein said Toll-like receptor ligand is E. coli K12 LPS.
10 . The method of claim 3 wherein the immune cells are neutrophils, the validated chemorepellent is a viral factor and wherein said viral factor is rvCMVUL146.
11 . The method of claim 3 wherein the immune cells are neutrophils, the validated chemorepellent is a semaphorin and wherein said semaphorin is semaphorin 3A.
12 . The method of claim 3 wherein the immune cells are neutrophils, the validated chemorepellent is an elastase inhibitor and wherein said elastase inhibitor is elafin.
13 . The method of claim 3 wherein the immune cells are neutrophils, the validated chemorepellent is a muscle cell proteins and wherein said muscle cell proteins is tropomyosin.
14 . The method of claim 3 wherein the wherein the immune cells are neutrophils, the validated chemorepellent is a plant cell wall component and wherein said plant cell wall component is citrus pectin.
15 . The method of claim 3 wherein the wherein the immune cells are neutrophils, the validated chemorepellent is an antibiotic and wherein said antibiotic is erythromycin.
16 . The method of claim 2 wherein the validated chemorepellent has a repellent index greater than 2.
17 . The method of claim 16 wherein the validated chemorepellent has a repellent index greater than 5.
18 . A method of screening for validated chemorepellents comprising:
a. obtaining a population of migratory cells,
b. contacting said population of migratory cells with a putative chemorepellent, wherein said putative chemorepellent is an agent suspected of having chemorepulsive activity for said migratory cells,
c. evaluating the migratory response of the population of migratory cells to the putative chemorepellent,
d. calculating the repellent index of the putative chemorepellent based on the migratory response in step (c), and
e. correlating those putative chemorepellents having a repellent index of greater than 1.2 with validated chemorepellents.
19 . The method of claim 18 wherein the migratory cells are neutrophils, T cells, B cells, monocytes or dendritic cells.
20 . The method of claim 19 wherein the neutrophils are obtained from a sample of anti-coagulated whole blood.
21 . A method of identifying cells which secrete at least one agent having chemorepulsive activity comprising:
a. preparing a conditioned media containing cells suspected of secreting at least one agent having chemorepulsive activity,
b. performing timed aliquot selection, wherein said timed aliquot selection comprises harvesting the supernatant from the cells of (a) at one or more timepoints between 1 and 360 days,
c. evaluating the migratory response of migratory cells to the supernatant harvested in (b) at each timepoint selected, wherein the supernatant harvested remains undiluted,
d. calculating the migration index of the undiluted supernatant from the migratory response in step (c) for each timepoint selected, and
e. correlating those aliquots having a migration index of greater than 1.2 with cells which secrete at least one agent having chemorepulsive activity.
22 . The method of claim 21 wherein cells suspected of secreting at least one agent having chemorepulsive activity is derived from a diseased tissue or organ.
23 . A method of inducing negative chemotaxis in human eosinophils comprising contacting said human eosinophils with a chemorepellent, said chemorepellent selected from the group consisting of galectin-1, galectin-2, heat shock protein 27, heat shock protein 40, heat shock protein 47, heat shock protein 70, heat shock protein 90, Pam3CSK4, HKLM (heat killed L. monocytogenes ), Poly(I:C), FLS-1, imiquimod, paclitaxel, Lipid A Diphos, P. gingavalis LPS, Lipomannan M. smegmatis , LTA Staph, Standard LTA from S. aureus , N-acetylmuramyl (MDP), cathespin G, resveratrol and quercetin dehydrate.
24 . A method of ameliorating an inflammatory response in a subject comprising:
a. identifying a subject having undergone an inflammatory insult or stimulus, and
b. contacting said subject with a validated chemorepellent at a site of inflammatory response to said inflammatory insult or stimulus in an amount effective to induce negative chemotaxis of at least a portion of a population of immune cells which comprise said inflammatory response.
25 . The method of claim 24 wherein said population of immune cells are neutrophils.