Anti-CD3 antibody-aminodextran conjugates for induction of T-cell activation and proliferation

• US 5,658,741 A
• Filed: 07/19/1995
• Issued: 08/19/1997
• Est. Priority Date: 06/11/1993
• Status: Expired due to Fees

• Current Assignee: Coulter International Corp.
• Original Assignee: Coulter Corporation (Danaher Corporation)
• Inventors: Kenyon, Norma Sue, Healy, Cynthia G., Bolton, Wade E., Maples, John A., Siiman, Olavi
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US08/504,095
• Time in Patent Office: 762 Days
• Field of Search: 435/6, 435/7.2, 435/72.4, 436/528, 436/529, 436/547, 436/548, 530/388.1, 530/391.1, 530/391.5, 530/388.73, 530/388.75
• US Class Current: 435/7.2
• CPC Class Codes: A61K 47/6849 the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
• CPC Class Codes: A61K 47/6883 Polymer-drug antibody conjugates, e.g. mitomycin-dextran-Ab; DNA-polylysine-antibody complex or conjugate used for therapy
• CPC Class Codes: G01N 33/531 Production of immunochemical test materials
• CPC Class Codes: G01N 33/56972 White blood cells
• CPC Class Codes: G01N 33/56988 HIV or HTLV

Abstract

The invention describes the use of novel aminodextran compounds containing about 5-20% by weight amine groups to bind a plurality of monoclonal antibodies. The resulting antibody-aminodextran compounds may be used to induce the activation and proliferation of selected mammalian cells. Specific examples are given using an anti-CD3 monoclonal antibody conjugated two aminodextrans containing about 5% and 16%, respectively, by weight amine groups as an agent for inducing the activation and proliferation of T cells.

Claims

1. A method of inducing the activation and proliferation of T-cells, said method comprising:

(a) conjugating a plurality of T-cell specific monoclonal antibodies to an aminodextran molecule having 7-20% by weight amine groups and a molecular weight of at least 100,000 daltons, wherein the molar ratio of said antibodies to said aminodextran is greater than or equal to two; and

(b) reacting said conjugate with a sample containing said T-cells to effect the binding of said conjugated antibodies to said T-cells to induce activation and proliferation of said T-cells.

2. The method according to claim 1 wherein said monoclonal antibody is specific to CD3 sites on T cells.

3. The method according to claim 2 wherein said aminodextran contain 15-17% by weight amine groups.

4. The method according to claim 1 wherein said aminodextran contains 10-17% by weight amine groups.

5. A method of analyzing mammalian T-cell function, said method comprising:

(a) reacting a sample containing mammalian T-cells with an aminodextran/anti-T-cell monoclonal antibody conjugate, said aminodextran having 7-20% by weight amine group and a molecular weight of at least 100,000 daltons, wherein the molar ratio of said antibodies to said aminodextran is greater than or equal to two; and

(b) analyzing product from step (a) to evaluate T-cell function by the use of at least one test selected from the group consisting of changes in nucleic acid synthesis, protein or glycoprotein synthesis, cellular size and morphology, membrane integrity, expression of cellular constituents, cell function, cell growth, cell differentiation and release of cellular components to the sample containing the cells.

6. The method according to claim 5 wherein said analysis is directed towards at least one disease or disorder selected from the group consisting of AIDS, non-AIDS immunodificiencies, infectious diseases, cancer, autoimmunity disorders and atopic diseases.

7. The method according to claim 6 wherein prior to step (b), non-T leukocyte cells and their immature precursors may be stimulated with selected agents to facilitate additional testing for said disease or disorder.

8. The method of claim 7 wherein said non-T leukocytes are selected from the group consisting of B cells, macrophages/monocytes, dendritic cells, neutrophils, eosinophils, basophils, cytotoxic effector cells, hematopoietic stem cells and immature precursor cells of each of these cells.

9. The method according to claim 5 wherein prior to step (b), non-T leukocyte cells and their immature precursors may be stimulated with selected agents to facilitate additional testing relating to said transplants.

10. The method according to claim 5 where the T-cells in said sample are obtained from a recipient of transplanted cells, tissues and organs.

11. The method of claim 5 wherein said aminodextran contains 10-17% by weight amine groups.

12. The method of claim 11 wherein said aminodextran contain 15-17% by weight amine groups.