EP0289949A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Intercellular adhesion molecules, and their binding ligands
Assignee: Dana Farber Cancer Institute Inc
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Filing: 1988-04-29
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Granted: —
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Expires: 2008-04-29
Abstract
Abstract The present invention relates to intercellular adhesion molecules (ICAM-1) which are involved in the process through which lymphocytes recognize and migrate to sites of inflammation as well as attach to cellular substrates during inflammation. The invention is directed toward such molecules, screening assays for identifying such molecules and antibodies capable of binding such molecules. The invention also includes uses for adhesion molecules and for the antibodies that are capable of binding them.
Inventors
- 👤 Timothy Alan Springer
- 👤 Robert Rothlein
- 👤 Steven Dean Marlin
- 👤 Michael Loran Dustin
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-26 06:00
CPC Classifications
C; C07; C07K; C07K16/00; C07K16/18; C07K16/28; C07K16/2803; C07K16/2821; A; A61; A61P; A61P29/00; A61P35/00; C07K14/00; C07K14/435; C07K14/705; C07K14/70503; C07K14/70525; C07K16/2839; C07K16/2845; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/575; G01N33/5751; A61K; A61K38/00; C07K2317/00; C07K2317/70; C07K2317/73
Claims Analysis
Total Claims: 36
Claim 1
Independent Claim
11 words
ICAM-1, or a functional derivative thereof, substantially free of natural contaminants.
Claim 2
Dependent Claim
23 words
The ICAM-1 of claim 1, wherein said ICAM-1 is additionally capable of binding to a molecule present on the surface of a lymphocyte.
Claim 3
Dependent Claim
56 words
The ICAM-1 molecule of claim 2, wherein said molecule additionally contains at least one polypeptide selected from the group consisting of: (a) -V-T-C-S-T-S-C-D-Q-P-K; (b) -X-G-S-V-L-V-T-C-S-T-S-C-D-Q-P-K; (c) -L-L-G-I-E-T-P-L; (d) -F-L-T-V-Y-X-T; (e) -V-E-L-A-P-L-P; (f) -E-L-D-L-R-P-Q-G-L-E-L-F-E; (g) -L-N-P-T-V-T-Y-G-X-D-S-F-S-A-K; (h) -S-F-P-A-P-N-V; (i) -L-R-G-E-K-E-L; (j) -R-G-E-K-E-L-K-R-E-P; (k) -L-R-G-E-K-E-L-K-R-E-P-A-V-G-E-P-A-E; (l) -P-R-G-G-S; (m) -P-G-N-N-R-K; (n) -Q-E-D-S-Q-P-M; (o) -T-P-E-R-V-E-L-A-P-L-P-S; (p) -R-R-D-H-H-G-A-N-F-S; and (q) -D-L-R-P-Q-G-L-E.
Claim 4
Dependent Claim
33 words
A functional derivative of the ICAM-1 of claim 2 wherein said functional derivative is a fragment of ICAM-1 which is capable of binding to a molecule present on the surface of a lymphocyte.
Claim 5
Dependent Claim
33 words
A functional derivative of the ICAM-1 of claim 2 wherein said functional derivative is a variant of ICAM-1 which is capable of binding to a molecule present on the surface of a lymphocyte.
Claim 6
Dependent Claim
33 words
A functional derivative of the ICAM-1 of claim 2 wherein said functional derivative is an analog of ICAM-1 which is capable of binding to a molecule present on the surface of a lymphocyte.
Claim 7
Dependent Claim
34 words
A functional derivative of the ICAM-1 of claim 2 wherein said functional derivative is a chemical derivative of ICAM-1 which is capable of binding to a molecule present on the surface of a lymphocyte.
Claim 8
Dependent Claim
13 words
A recombinant DNA molecule capable of expressing ICAM-1 or a functional derivative thereof.
Claim 9
Dependent Claim
63 words
The DNA molecule of claim 8, wherein said ICAM-1 or said functional derivative thereof is capable of encoding at least one polypeptide selected from the group consisting of: (a) -V-T-C-S-T-S-C-D-Q-P-K; (b) -X-G-S-V-L-V-T-C-S-T-S-C-D-Q-P-K; (c) -L-L-G-I-E-T-P-L; (d) -F-L-T-V-Y-X-T; (e) -V-E-L-A-P-L-P; (f) -E-L-D-L-R-P-Q-G-L-E-L-F-E; (g) -L-N-P-T-V-T-Y-G-X-D-S-F-S-A-K; (h) -S-F-P-A-P-N-V; (i) -L-R-G-E-K-E-L; (j) -R-G-E-K-E-L-K-R-E-P; (k) -L-R-G-E-K-E-L-K-R-E-P-A-V-G-E-P-A-E; (l) -P-R-G-G-S; (m) -P-G-N-N-R-K; (n) -Q-E-D-S-Q-P-M; (o) -T-P-E-R-V-E-L-A-P-L-P-S; (p) -R-R-D-H-H-G-A-N-F-S; and (q) -D-L-R-P-Q-G-L-E.
Claim 10
Independent Claim
91 words
A method for recovering ICAM-1 in substantially pure form which comprises the steps: (a) solubilizing ICAM-1 from the membranes of cells expressing ICAM-1, to form a solubilized ICAM-1 preparation, (b) introducing said solubilized ICAM-1 preparation to an affinity matrix, said matrix containing immobilized antibody capable of binding to ICAM-1, (c) permitting said ICAM-1 to bind to said antibody of said affinity matrix, (d) removing from said matrix any compound incapable of binding to said antibody and (e) recovering said ICAM-1 in substantially pure form by eluting said ICAM-1 from said matrix.
Claim 11
Dependent Claim
37 words
The method of claim 10 wherein said method additionally comprises the steps: (f) purifying said recovered ICAM-1 of step (e) by preparative gel electrophoresis, and (g) eluting said recovered ICAM-1 from a gel employed in step (f).
Claim 12
Dependent Claim
12 words
The ICAM-1 produced by the method of any one of claims 10-11.
Claim 13
Dependent Claim
21 words
An antibody capable of binding to a molecule selected from the group consisting of ICAM-1, and a functional derivative of ICAM-1.
Claim 14
Dependent Claim
12 words
The antibody of claim 13, wherein said antibody is a monoclonal antibody.
Claim 15
Dependent Claim
9 words
The monoclonal antibody of claim 14, which is R6-5-D6.
Claim 16
Dependent Claim
47 words
The antibody of claim 13 wherein said molecule is capable of binding to a receptor present on the surface of a lymphocyte, and wherein the binding of said antibody to said molecule impairs the ability of said molecule to bind to said receptor molecule of said lymphocyte.
Claim 17
Dependent Claim
12 words
The antibody of claim 16, wherein said antibody is a monoclonal antibody.
Claim 18
Dependent Claim
11 words
The antibody of any one of claims 13-17, in labeled form.
Claim 19
Dependent Claim
18 words
A hybridoma cell capable of producing the monoclonal antibody of any one of claims 14, 15, and 17.
Claim 20
Dependent Claim
16 words
The hybridoma cell capable of producing the monoclonal antibody R6-5-D6, said cell being ATCC HB 9580.
Claim 21
Dependent Claim
19 words
A fragment of the antibody of any one of claims 13-17, said fragment being capable of binding said molecule.
Claim 22
Independent Claim
86 words
A method for producing a desired hybridoma cell that produces an antibody which is capable of binding to ICAM-1, or its functional derivative, which comprises the steps: (A) immunizing an animal with a cell expressing ICAM-1, (B) fusing the spleen cells of said animal with a myeloma cell line, (C) permitting the fused spleen and myeloma cells to form antibody secreting hybridoma cells, and (D) screening said hybridoma cells for said desired hybridoma cell that is capable of producing an antibody capable of binding to ICAM-1.
Claim 23
Dependent Claim
103 words
The method of claim 22, wherein in step (A) said animal is immunized with a cell expressing ICAM-1 but not expressing LFA-1, and wherein said screening step (D) comprises the steps: (1) incubating the antibody secreted from any of said hybridoma cells with a lymphocyte preparation, said lymphocyte preparation containing a plurality of cells capable of aggregating, (2) examining said secreted antibody for the capacity to inhibit the aggregation of said cells of said lymphocyte preparation, and (3) selecting as said desired hybridoma cell a hybridoma cell that produces an antibody capable of inhibiting said aggregation of said cells of said lymphocyte preparation.
Claim 24
Dependent Claim
130 words
The method of claim 22 wherein said screening step (D) comprises the steps: (1) incubating the antibody secreted from any of said hybridoma cells with: a lymphocyte preparation containing a plurality of cells having the characteristics of: (a) being capable of aggregating, and (b) being incapable of aggregating in the presence of an antibody capable of binding ICAM-1, (2) verifying that said antibody secreted from said hybridoma cell does not bind to a member of the LFA-1 family of molecules, (3) selecting as said desired hybridoma cell a hybridoma cell that produces an antibody capable of inhibiting the spontaneous aggregation of the cells of said lymphocyte preparation, and (4) verifying that said antibody selected from said hybridoma cell does not bind to a member of the LFA-1 family of molecules.
Claim 25
Dependent Claim
13 words
The hybridoma cell obtained from the method of any one of claims 22-24.
Claim 26
Dependent Claim
10 words
The antibody produced by the hybridoma cell of claim 25.
Claim 27
Independent Claim
77 words
A method of identifying a non-immunoglobulin antagonist of intercellular adhesion which comprises: (A) incubating a non-immunoglobulin agent capable of being an antagonist of intercellular adhesion with a lymphocyte preparation, said lymphocyte preparation containing a plurality of cells capable of aggregating, (B) examining said lymphocyte preparation to determine whether the presence of said agent inhibits the aggregation of said cells of said lymphocyte preparation; wherein inhibition of said aggregation identifies said agent as an antagonist of intercellular adhesion.
Claim 28
Independent Claim
74 words
A method of diagnosing the presence and location of an ICAM-1-expression tumor cell in a mammalian subject suspected of having such a cell, which comprises: a) administering to said subject a composition containing a detectably labeled binding ligand capable of binding to ICAM-1, said ligand being selected from the group consisting of an antibody and a fragment of an antibody, said fragment being capable of binding to ICAM-1, and b) detecting said binding ligand.
Claim 29
Independent Claim
62 words
A method of diagnosing the presence and location of inflammation resulting from a response of the specific defence system in a mammalian subject suspected of having said inflammation which comprises a) incubating a sample of tissue of said subject with a composition containing a detectably labeled binding ligand capable of identifying a cell which expresses ICAM-1, and b) detecting said binding ligand.
Claim 30
Independent Claim
24 words
The method of any one of claims 28 or 29, wherein said binding ligand is bound to ICAM-1 present in said sample of tissue.
Claim 31
Dependent Claim
45 words
The binding ligand of any one of claims 28 or 29, wherein said binding ligand is selected from the group consisting of: a monoclonal antibody capable of binding to ICAM-1; and a fragment of said monoclonal antibody, said fragment being capable of binding to ICAM-1.
Claim 32
Dependent Claim
15 words
The binding ligand of claim 31, wherein said monoclonal antibody is the monoclonal antibody R6-5-D6.
Claim 33
Independent Claim
80 words
A method of diagnosing the presence and location of a tumor cell which expresses a member of the LFA-1 family of molecules in a subject suspected of having such a cell, which comprises: a) administering to said subject a composition containing a detectably labeled binding ligand capable of binding to a member of the LFA-1 family of molecules, said ligand being selected from the group consisting of ICAM-1 and a functional derivative of ICAM-1 and b) detecting said binding ligand.
Claim 34
Independent Claim
106 words
A method of diagnosing the presence and location of a tumor cell which expresses a member of the LFA-1 family of molecules in a subject suspected of having such a cell, which comprises: a) incubating a sample of tissue of said subject in the presence of a composition containing a detectably labeled binding ligand capable of binding to a member of the LFA-1 family of molecules, said ligand being selected from the group consisting of ICAM-1 and a functional derivative of ICAM-1 and b) detecting said binding ligand which is bound to a member of the LFA-1 family of molecules present in said sample of tissue.
Claim 35
Dependent Claim
73 words
A pharmaceutical composition comprising ICAM-1, a functional derivative of ICAM-1, an antibody or toxin derivitized antibody capable of binding to a molecule selected from ICAM-1 and a functional derivative of ICAM-1, a fragment of an antibody or toxin derivitized fragment of an antibody, said fragment being capable of binding to a molecule selected from ICAM-1 and a functional derivative of ICAM-1, or a non-immunoglobulin antagonist of ICAM-1, together with an inert pharmaceutical carrier.
Claim 36
Dependent Claim
103 words
ICAM-1, a functional derivative of ICAM-1, an antibody or toxin derivitized antibody capable of binding to a molecule selected from ICAM-1 and a functional derivative of ICAM-1, a fragment of an antibody or toxin derivitized fragment of an antibody, said fragment being capable of binding to a molecule selected from ICAM-1 and a functional derivative of ICAM-1, or a non-immunoglobulin antagonist of ICAM-1, for use in the preparation of a pharmaceutical composition for treating inflammation resulting from a response of the specific defense system of a mammalian subject, or for supressing the growth of an ICAM-1 expressing tumor or an LFA-1 expressing tumor.
References Cited (Backward Citations)
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