Monoclonal antibodies and their use in a method for monitoring subsets of activated T cells

Claim 1

A method for monitoring subpopulations of activated T lymphocytes in a sample comprising the steps of: (a) isolating a cell sample containing activated T lymphocytes; (b) combining said cell sample with a first monoclonal antibody selected from the group consisting of anti L-35 (ATCC HB 9050) and anti L-36 (ATCC HB 9051), said first monoclonal antibody being conjugated to a first fluorescent dye, and with a second monoclonal antibody selected from the group consisting of anti-CD5, anti-CD8, anti-CD4 and anti-CD3 monoclonal antibodies, said second monoclonal antibody being conjugated to a second fluorescent dye, said first and second fluorescent dyes having emission wavelengths which differ a sufficient amount to produce separately detectable signals, wherein said first and second monoclonal antibodies bind to said activated T lymphocytes; (c) detecting fluorescence emission from said first and second fluorescent dyes; and (d) monitoring subpopulations of activated T lymphocytes in said sample as a function of said detected fluorescence emission.

Claim 2

A method for monitoring subpopulations of activated T lymphocytes in a sample comprising the steps of: (a) isolating a cell sample containing activated T lymphocytes; (b) combining said cell sample with a first monoclonal antibody selected from the group consisting of anti L-35 (ATCC HB 9050) and anti L-36 (ATCC HB 9051), said first monoclonal antibody being conjugated to a first fluorescent dye, and with a second monoclonal antibody selected from the group consisting of anti-CD5, anti-CD8, anti-CD4 and anti-CD3 monoclonal antibodies, said second monoclonal antibody being conjugated to a second fluorescent dye, said first and second fluorescent dyes having emission wavelengths which differ a sufficient amount to produce separately detectable signals, wherein said first and second monoclonal antibodies bind to said activated T lymphocytes; (c) detecting fluorescence emission from said first and second fluorescent dyes; and (d) monitoring subpopulations of activated T lymphocytes in said sample as a function of said detected fluorescence emission.

Claim 8

A method for monitoring subpopulations of activated leukocytes in a cell sample comprising the steps of: (a) combining a cell sample containing activated leukocytes with a monoclonal antibody conjugated to a detectable label, said antibody being selected from the group consisting of anti L-35 (ATCC HB 9050) and anti L-36 (ATTCC HB 9051), where said monoclonal antibody binds to said activated leukocytes; (b) causing said label to provide a detectable signal; (c) detecting said signal; and (d) monitoring subpopulations of activated leukocytes in said cell sample by characteristics of said signal.

Claim 9

A method for monitoring subpopulations of activated leukocytes in a cell sample comprising the steps of: (a) combining a cell sample containing activated leukocytes with a monoclonal antibody conjugated to a detectable label, said antibody being selected from the group consisting of anti L-35 (ATCC HB 9050) and anti L-36 (ATTCC HB 9051), where said monoclonal antibody binds to said activated leukocytes; (b) causing said label to provide a detectable signal; (c) detecting said signal; and (d) monitoring subpopulations of activated leukocytes in said cell sample by characteristics of said signal.

Claim 14

A method for monitoring subpopulations of activated human T lymphocytes in a sample comprising the steps of: (a) isolating a cell sample containing activated T lymphocytes from a human blood sample; (b) combining said cell sample with a monoclonal antibody selected from the group consisting of anti L-35 (ATTCC HA 9050) and anti L-36 (ATCC HB 9051), said antibody being conjugated to fluorescein, and with an anti-CD3 monoclonal antibody, said anti-CD3 being conjugated to phycoerythrin B, whereby said antibodies bind to said activated T lymphocytes to give labeled T lymphocytes; (c) detecting two-color fluorescence emission from said labeled T lymphocytes; and (d) monitoring subpopulations of activated T lymphocytes in said blood sample as a function of said detected two-color fluorescence emission.

Claim 15

A method for monitoring subpopulations of activated human T lymphocytes in a sample comprising the steps of: (a) isolating a cell sample containing activated T lymphocytes from a human blood sample; (b) combining said cell sample with a monoclonal antibody selected from the group consisting of anti L-35 (ATTCC HA 9050) and anti L-36 (ATCC HB 9051), said antibody being conjugated to fluorescein, and with an anti-CD3 monoclonal antibody, said anti-CD3 being conjugated to phycoerythrin B, whereby said antibodies bind to said activated T lymphocytes to give labeled T lymphocytes; (c) detecting two-color fluorescence emission from said labeled T lymphocytes; and (d) monitoring subpopulations of activated T lymphocytes in said blood sample as a function of said detected two-color fluorescence emission.

Claim 16

A method for monitoring subpopulations of activated T lymphocytes in a body tissue comprising the steps of: (a) isolating a sample of a body tissue containing activated T lymphoctyes having antigens expressed on the surfaces thereof; (b) adding to said tissue a first monoclonal antibody selected from the group consisting of anti L-35 (ATCC HB 9050) and anti L-36 (ATCC HB 9051), said first antibody being conjugated to a first fluorescent dye, and a second monoclonal antibody selected from the group consisting of anti-CD5, anti-CD8, anti-CD4 and anti-CD3 monoclonal antibodies, said second antibody being conjugated to a second fluorescent dye, said first and second dyes having emission wavelengths which differ by at least 25 nm, wherein said antigens on said activated T lymphocytes bind to said first and second antibodies to give labeled T lymphocytes; (c) subjecting said labeled T lymphocytes to excitation light to cause fluorescence from said labeled T lymphocytes; (d) detecting fluorescence emission from said labeled T lymphocytes; and (e) monitoring subpopulations of activated T lymphocytes in said tissue as a function of said detected fluorescence emission.

Claim 17

A method for monitoring subpopulations of activated T lymphocytes in a body tissue comprising the steps of: (a) isolating a sample of a body tissue containing activated T lymphoctyes having antigens expressed on the surfaces thereof; (b) adding to said tissue a first monoclonal antibody selected from the group consisting of anti L-35 (ATCC HB 9050) and anti L-36 (ATCC HB 9051), said first antibody being conjugated to a first fluorescent dye, and a second monoclonal antibody selected from the group consisting of anti-CD5, anti-CD8, anti-CD4 and anti-CD3 monoclonal antibodies, said second antibody being conjugated to a second fluorescent dye, said first and second dyes having emission wavelengths which differ by at least 25 nm, wherein said antigens on said activated T lymphocytes bind to said first and second antibodies to give labeled T lymphocytes; (c) subjecting said labeled T lymphocytes to excitation light to cause fluorescence from said labeled T lymphocytes; (d) detecting fluorescence emission from said labeled T lymphocytes; and (e) monitoring subpopulations of activated T lymphocytes in said tissue as a function of said detected fluorescence emission.