Chemistry: natural resins or derivatives; peptides or proteins; – Peptides of 3 to 100 amino acid residues
Reexamination Certificate
1995-06-05
2001-04-10
Romeo, David S. (Department: 1647)
Chemistry: natural resins or derivatives; peptides or proteins;
Peptides of 3 to 100 amino acid residues
C530S350000, C530S351000, C435S069100, C435S069500, C514S002600, C424S085100
Reexamination Certificate
active
06214968
ABSTRACT:
BACKGROUND OF THE INVENTION
The field of the invention is lymphokines and fibrosis.
Schistosomiasis is one of the most important helminthic diseases, estimated to afflict 200 million people in the tropics (Walsh et al., 1979). Two of the schistosome species that infect humans (
Schistosoma mansoni
and
S. japonicum
) can cause serious morbidity (including portal hypertension and gastrointestinal hemorrhage) as a result of a form of hepatic fibrosis (Cheever et al., 1967). However, only a relatively small subpopulation (3-6%) of infected individuals develop this scarring; the others remain generally healthy (Chen et al., 1988).
Traditional forms of antihelminthic therapy have a number of undesirable side effects, and treatment with the relatively new drug, praziquantel, is very expensive, thus making antihelminthic treatment of all infected individuals impractical. In addition, while early antihelminthic therapy may aid in preventing liver scarring, it has not been established that this is an invariable outcome of treatment (Homeida et al., 1988).
Alternatively, early aggressive treatment of infected individuals with anti-inflammatory or immunosuppressive drugs including methotrexate, cytotoxins and various corticosteriods may aid the prevention of scarring. However, given that these drugs are known to produce a number of relatively severe side effects, treatment of all infected individuals, 94-97% of which would never develop the progressive fibrotic form of the disease, is both undesirable and impractical.
Several other chronic inflammatory diseases, including pulmonary fibrosis, scleroderma/progressive systemic sclerosis, sarcoidosis, sclerosing cholangitis, primary biliary cirrhosis and inflammatory bowel disease also can result in organ dysfunction due to pathological fibrosis. As in schistosomiasis, only a subpopulation of individuals with these diseases develop debilitating organ scarring. Thus, methods are needed that would make it possible to predict which patients will develop the progressive fibrotic forms of these inflammatory diseases so that more aggressive, antifibrotic courses of treatment can be limited to only those individuals who would benefit from these treatments.
SUMMARY OF THE INVENTION
In general, the invention features a method for identifying individuals with a propensity for pathological fibrosis. The method comprises providing a sample from an individual with a chronic inflammatory disease, contacting the sample with an antibody specific for FsF-1 under conditions which permit immunocomplex formation, and detecting an increase in the relative level of the immunocomplex as an indication of a propensity for pathological fibrosis. By “relative level” is meant the relative amount of immunocomplex detected when compared to the level in a sample from a normal individual.
In an individual at a chronic stage of the disease, an increased level of the immunocomplex in a single sample is indicative that they are at risk of serious fibrosis. In an individual at an early stage of the disease, the method further involves providing serial samples from the individual over a period of time (e.g., every 3 to 6 months over a period of 1 to 3 years), and detecting a persistent increase in the relative level of the immunocomplex as an indication of a propensity for pathological fibrosis.
The sample may be any biological sample. Preferably, the sample is a blood, serum or plasma sample, but may also be a urine sample; a tissue sample (e.g., biopsy); an effusion obtained from a joint, the abdominal cavity (e.g., ascites), pleural fluid, cerebral spinal fluid, and the aqueous humor; or from the supernatant of cultured peripheral blood mononuclear cells. Also preferably, the sample is obtained from a mammal, and even more preferably, the mammal is a human.
In one preferred embodiment, the pathological scarring results from hepatic fibrosis. In another related embodiment, hepatic fibrosis is the result of the disease schistosomiasis. In other embodiments, the pathological fibrosis is a result of various chronic inflammatory diseases including sarcoidosis, scleroderma, sclerosing cholangitis, rheumatoid arthritis, pulmonary fibrosis, and interstitial pneumonitis.
The invention also features a substantially pure fibroblast stimulating factor-1 (FsF-1) polypeptide. By “FsF-1 polypeptide” is meant all or part of a novel lymphokine, also referred to as fibrosin, that is a heparin-binding growth factor which stimulates fibroblast proliferation, collagen and hyaluronan synthesis and fibroblast chemotaxis, and which is distinct from other previously characterized heparin-binding growth factors. Preferably, FsF-1 is produced in CD4
+
lymphocytes, and the naturally occurring intact polypeptide is characterized as being of molecular weight of about 60 kD as measured by SDS-PAGE, and being of isoelectric point of about 6.2. By “polypeptide” is meant any chain of amino acids, regardless of length or post-translational modification (e.g., glycosylation).
A further feature of the invention is a substantially pure antibody which specifically binds FsF-1. By “specifically binds” is meant an antibody which binds to FsF-1 and which does not substantially recognize and bind to other antigenically-unrelated molecules. Antibodies according to the invention may be prepared by a variety of methods. For example, the FsF-1 protein or antigenic fragments thereof can be administered to an animal in order to induce the production of polyclonal antibodies. Alternatively, antibodies according to the invention may be monoclonal antibodies. Such monoclonal antibodies can be prepared using hybridoma technology (see, e.g., Kohler et al.,
Nature
256:495, 1975; Kohler et al.,
Eur. J. Immunol.
6:511, 1976; Kohler et al.,
Eur J. Immunol.
6:292, 1976; Hammerling et al.,
In Monoclonal Antibodies and T Cell Hybridomas,
Elsevier, N.Y., 1981).
As used herein, the term “substantially pure” describes a compound, e.g., a protein, polypeptide, or antibody, that is substantially free from the components that naturally accompany it. Typically, a compound is substantially pure when at least 60%, more preferably at least 75%, more preferably at least 90%, and most preferably at least 99%, of the total material (by weight) in a sample is the compound of interest. Purity can be measured by any appropriate method, e.g., in the case of polypeptides by column chromatography, polyacrylamide gel electrophoresis, or HPLC analysis.
The FsF-1 polypeptide, according to the invention, may be used as the active ingredient of therapeutic compositions. In such therapeutic compositions, the active ingredient may be formulated with a physiologically-acceptable carrier or anchored in the membrane of a cell. Such therapeutic compositions are used to stimulate fibroblast proliferation and extracellular matrix synthesis, e.g., to promote wound healing. The method involves applying the therapeutic composition, preferably topically, to a wound of a mammal in a dosage effective to stimulate fibroblast proliferation and thereby accelerate wound closure.
In another aspect, the invention also features isolated DNA consisting essentially of a DNA sequence encoding an FsF-1 polypeptide of a vertebrate animal, preferably a mammal, which polypeptide has an amino acid sequence with at least 50% (preferably 60%, more preferably at least 70%, and most preferably at least 85%) homology to the amino acid sequence encoded by the nucleotide sequence shown in
FIG. 18
(SEQ ID NO.: 1),
FIG. 25
(SEQ ID NO.: 2) or
FIG. 26
(SEQ ID NO.: 3).
By “isolated”, as used herein in reference to DNA, is meant a DNA that is not immediately contiguous with (i.e., covalently linked to) both of the coding sequences with which it is immediately contiguous in the naturally occurring genome of the organism from which the DNA of the invention is derived. The term therefore includes, for example, a recombinant DNA which is incorporated into a vector (e.g., an autonomously replicating virus or plasmid), or into the genomic DNA of a prokaryote or eukaryote; DNA which e
Prakash Sadhana
Wyler David J.
Zhang Xiaoping
Fish & Richardson P.C.
New England Medical Center Hospitals Inc.
Romeo David S.
LandOfFree
Fibroblast stimulating growth factor 1 (FsF-1) and the early... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Fibroblast stimulating growth factor 1 (FsF-1) and the early..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Fibroblast stimulating growth factor 1 (FsF-1) and the early... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2443383