Compositions and methods for detecting and treating pathological fibroblast cells
a technology of pathological fibroblasts and compositions, applied in the field of compositions and methods for detecting and treating pathological fibroblasts, can solve the problems of lack of current existence, loss of function, organ failure, etc., and achieve the effect of reducing the number of pathologic myofibroblast subpopulations, rapid elimination of this pathologic myofibroblast subpopulation, and inducing cell death
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[0237]The following examples are illustrative, but not limiting, of the compounds, compositions, and methods of the present invention. Other suitable modifications and adaptations of the variety of conditions and parameters normally encountered in clinical therapy and which are obvious to those skilled in the art are within the spirit and scope of the invention.
example i
[0238]This example describes the expression of Pro-N-cadherin on the cell surface of fibroblasts isolated from tissues pathological in origin but not expressed on the cell surface of fibroblasts isolated from non-diseased tissues.
[0239]A murine monoclonal antibody (mAb) was obtained from the University of Nebraska Medical School. This antibody is highly specific for the precursor (pro) domain of Pro-N-cadherin (see, Wahl, J. K., et al Journal of Biological Chemistry, 2003. 278(19): p. 17269-17276). Western blot analysis shows that this mAb recognizes Pro-N-cadherin protein from human pathological fibroblasts from lung (LL97A), heart (CF-DCM) and liver (LX2) with no protein expression detected in normal fibroblasts NHLF and CCD-16lu (FIG. 1B) due to normal N-cadherin protein processing. Aberrant cell surface localization of Pro-N-cadherin is demonstrated by flow cytometry analyzed pathological fibroblasts from lung (LL97A), heart (CF-DCM) and liver (LX2) with no cell surface localiza...
example ii
[0240]This example demonstrates that mAb treatment targeting the pro-domain of the precursor N-cadherin results in reduction of proliferation, activation and viability of pathological fibroblasts. This example also demonstrates the phenomenon defined as the hook effect (Also known as the prozone effect) related to monoclonal antibodies.
[0241]To assess whether administering an antibody against the Pro-N-cadherin protein can result in cell death, LL97A diseased fibroblasts were treated monoclonal antibody (10A10), that specifically targets the pro domain of the precursor N-cadherin molecule. Administration of the 10A10 antibody resulted in cell death of approximately seventy-five percent of the fibroblast population overnight, confirmed by trypan blue staining (FIG. 2). FIG. 2 demonstrates reduced viability of pathological LL97A fibroblasts with Pro-N-cadherin treatment. Pathological fibroblasts from heart, lung and liver were challenged with Pro-N-cadherin mAb and effects were measur...
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