Methods for determining T-cell profiles of immunocompromised subjects

• US 5,968,755 A
• Filed: 09/26/1996
• Issued: 10/19/1999
• Est. Priority Date: 09/27/1995
• Status: Expired due to Term

• Current Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford Management Co.)
• Original Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford Management Co.)
• Inventors: Rabin, Ronald, Herzenberg, Leonore A., Herzenberg, Leonard A., Roederer, Mario
• Primary Examiner(s): Huff, Sheela
• Application Number:
  US08/721,260
• Time in Patent Office: 1,118 Days
• Field of Search: 435/5, 435/7.1, 435/7.24, 530/387.1
• US Class Current: 435/7.24
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/505 involving T-cells
• CPC Class Codes: G01N 33/5094 for blood cell populations red blood cells G01N33/80
• CPC Class Codes: G01N 33/56972 White blood cells

Abstract

A method of identifying an abnormal T-cell profile of an immunocompromised subject is disclosed. The method involves detecting the immunoreactivity of T-cells with at least two antibodies selectively reactive with naive T-cell surface proteins and comparing data from cells isolated from an immunocompromised subject with data from cells isolated form a non-immunocompromised subject. Also disclosed are methods of screening drugs or compounds for efficacy to stimulate the production of naive T cells in a subject.

Claims

1. A method of measuring the T-cell profile of an immunocompromised subject relative to the T-cell profile of a non-immunocompromised subject, comprising

sorting a sample containing peripheral blood mononuclear cells (PBMC) obtained from said immunocompromised subject into one or more sets of T-cells types selected from the group consisting of CD4+, CD8+, and CD4+ plus CD8+,

determining in each set the ratio of naive cells relative to the total number of cells (naive:

total) by detecting immunoreactivity of T-cells in each set with at least two different antibodies selectively reactive with T-cell surface proteins, said antibodies comprising

a) a first antibody selectively reactive with a CD45RA T-cell surface protein, and

b) a second antibody selectively reactive with either a CD62L or a CD11a T-cell surface protein,

and comparing the naive;

(otal ratios of sets from said immunocompromised subject with the naive;

total ratios of analogous sets from a non-immunocompromised subject.

2. The method of claim 1, wherein the T-cell profile of the immunocompromised subject includes the preferential loss of naive T-cells in the immunocompromised subject.

3. The method of claim 1 applied to an HIV-infected subject.

4. The method of claim 1 applied to a subject undergoing immunotherapy.

5. The method of claim 1, wherein said determining is accomplished using a fluorescence-activated cell sorter (FACS) and three-color immunophenotyping.

6. The method of claim 1, wherein said first antibody is anti-CD45RA and said second antibody is anti-CD62L.

7. The method of claim 1, wherein said first antibody is anti-CD45RA and said second antibody is anti-CD11a.

8. A method of evaluating the efficacy of a drug to stimulate the production of naive T cells in a subject, comprising

obtaining a first sample containing peripheral blood mononuclear cells (PBMC) from said subject,

administering a suitable dose of said drug to said subject,

obtaining a second sample containing peripheral blood mononuclear cells (PBMC) from said subject,

isolating from said first and second samples, first and second populations of T cells, respectively,

determining the number of naive T cells in each population, said determining including the detection of immunoreactivity of T-cells in the populations with at least two different antibodies selectively reactive with T-cell surface proteins, said antibodies comprising

a) a first antibody selectively reactive with a CD45RA T-cell surface protein, and

b) a second antibody selectively reactive with either a CD62L or a CD11a T-cell surface protein, and

identifying the drug as effective if the number of naive T-cells in the second population is significantly greater than the number of naive T-cells in the first population.

9. The method of claim 8, wherein said isolating includes sorting each population into one or more sets of T-cell types selected from the group consisting of CD4+, CD8+, and CD4+ plus CD8+, and said determining includes determining the number of naive T cells in each set of each population.

10. The method of claim 8, wherein said first antibody is anti-CD45RA and said second antibody is anti-CD62L.

11. The method of claim 8, wherein said first antibody is anti-CD45RA and said second antibody is anti-CD11a.