Methods of detecting and treating vaso-occlusive crisis in sickle cell disease

• US 5,669,396 A
• Filed: 01/16/1996
• Issued: 09/23/1997
• Est. Priority Date: 01/16/1996
• Status: Expired due to Fees

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Bageac, Alexandru Cristian, Thatte, Hemant Sadashiv, Golan, David Eric
• Primary Examiner(s): Millin, Vincent
• Assistant Examiner(s): O'Hara, Kelly R.
• Application Number:
  US08/585,698
• Time in Patent Office: 616 Days
• Field of Search: 128/898, 424/529, 436/66, 436/547, 436/548, 436/811, 514/814, 514/815
• US Class Current: 128/898
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07K 16/2806 against CD2
• CPC Class Codes: C07K 16/2824 against CD58
• CPC Class Codes: G01N 33/491 by separating the blood components G01N15/05 takes precedence
• CPC Class Codes: G01N 33/56972 White blood cells
• CPC Class Codes: G01N 33/80 involving blood groups or blood types or red blood cells white blood cells G01N33/56972

Abstract

Methods are described for treating sickle cell disease, and in particular, detection and treatment of vaso-occlusive crisis in sickle cell disease. Antibodies are employed which bind competitively and suppress adhesion of sickle erythrocytes to autologous lymphocytes.

Claims

1. A method of testing blood, comprising:

a) providing blood from a patient with sickle cell disease; and

b) examining said blood to determined the incidence of preformed lymphocyte/erythrocyte conjugates as a test for sickle cell crisis.

2. The method of claim 1, wherein prior to said examining, said blood is subjected to density centrifugation under conditions such that a cell layer is generated comprising lymphocytes.

3. The method of claim 2, wherein after said centrifugation and prior to said examining, at least a portion of said lymphocytes is removed from said cell layer.

4. The method of claim 3, wherein said portion is placed on a microscope slide so that said examining is performed microscopically.

5. The method of claim 4, wherein the number of lymphocytes with at least one adherent erythrocyte are counted.

6. A method of testing blood, comprising:

a) providing i) erythrocytes from a patient with sickle cell disease; and

ii) autologous lymphocytes from said patient with sickle cell disease;

b) mixing said autologous lymphocytes and said erythrocytes in vitro to create a cell suspension under conditions such that lymphocyte/erythrocyte conjugates can form; and

c) determining the incidence of lymphocyte/erythrocyte conjugates as a test for sickle cell crisis.

7. The method of claim 6, wherein, prior to said mixing, said lymphocytes are exposed in vitro to antibody.

8. The method of claim 7, wherein said antibody comprises anti-CD2 antibody.

9. The method of claim 6, wherein, prior to said mixing, said erythrocytes are exposed in vitro to antibody.

10. The method of claim 9, wherein said antibody comprises anti-CD58 antibody.

11. The method of claim 6, wherein said autologous lymphocytes and said erythrocytes are mixed in cell culture media at the ratio of approximately 1 lymphocyte to 100 erythrocytes.

12. The method of claim 11, wherein said conditions comprise incubation of said cell suspension at approximately 37°

C.

13. The method of claim 12, wherein said incubation is carried out for between approximately one minute and twenty-four hours.

14. The method of claim 13, wherein said incubation is carried out for approximately one hour.

15. The method of claim 12, wherein after said incubation, said cell suspension is examined for lymphocyte/erythrocyte conjugates.

16. The method of claim 15, wherein, prior to said examining, at least a portion of said cell suspension is removed and placed on a microscope slide.

17. The method of claim 16, wherein said examining is performed microscopically and the number of lymphocytes with at least one adherent erythrocyte are counted.