Method for detecting CD4+CD25+glycoprotein A33(GPA33)high and/or CD4+CD127-GPA33high T cells
Inventors
AMSEN, Derk • CUADRADO GODIA, Eloy • Borst, Jannetje Geertruida
Assignees
Stichting Het Nederlands Kanker Institmit Antoni Van Leeuwenhoek Ziekenhuis • Netherlands Cancer Institute • Stichting Sanquin Bloedvoorziening
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Publication Number
US-12180458-B2
Publication Date
2024-12-31
Expiration Date
Abstract
The invention relates to isolated populations of CD4+GPA33high regulatory T cells, to methods for detecting and/or isolating CD4+GPA33high cells, methods for enriching for CD4+GPA33high cells and to uses of CD4+GPA33high cells and populations of such cells. The invention further relates to methods for classifying an individual suffering from cancer and typing a tumor sample as well as for determining the etiology of autoimmune diseases.
Core Innovation
The invention identifies glycoprotein A33(GPA33) as a cell-surface marker for stable CD4+ regulatory T cells (tTregs). It relates GPA33 expression to CD4+ regulatory T cell stability and supports using GPA33 to detect, enrich, and isolate CD4+GPA33high Tregs. The described detection and selection also can incorporate CD25 and CD127 characteristics to define the relevant CD4+ Treg subpopulations.
The invention provides methods to detect CD4+CD25+GPA33high and/or CD4+CD127−GPA33high T cells in a sample by contacting cells with an anti-CD4 antibody or antigen-binding fragment thereof and an anti-GPA33 antibody or antigen-binding fragment thereof to detect binding, and further contacting the sample with an anti-CD25 antibody and/or an anti-CD127 antibody to detect binding or the lack thereof for defining the T cell subset. GPA33high is defined by GPA33 expression that is higher than an average level of GPA33 expression on CD4+ T cells in the cell sample, and is further described with quantitative alternative definitions in the claim set.
The invention further supports use of CD4+GPA33high Tregs for adoptive immunotherapy, including improved stability and suppressive function after expansion without rapamycin. It also links GPA33-based CD4+ Treg detection to clinical use cases in autoimmune disease and cancer by quantifying CD4+GPA33high cells, optionally with CD25/CD127 patterns, to classify or prognose disease and to predict therapy response by comparing tumor samples to reference samples.
Claims Coverage
The provided claim set includes two independent methods. Both independent claims cover antibody-based detection of CD4+ regulatory T-cell subsets defined by GPA33high and, optionally, CD25 positivity and/or CD127− status, with GPA33high defined by an expression threshold criterion.
Antibody-based multi-marker detection of CD4+ and GPA33high T cells
Contacting a cell sample with an anti-CD4 antibody or antigen-binding fragment thereof and an anti-GPA33 antibody or antigen-binding fragment thereof and detecting binding between cells in the sample and the anti-CD4 and anti-GPA33 antibodies or binding fragments to detect CD4+CD25+GPA33high and/or CD4+CD127−GPA33high T cells.
CD25 binding and CD127 binding-or-lack detection for subset discrimination
Contacting the cell sample with an anti-CD25 antibody or antigen-binding fragment thereof and detecting binding between cells in the sample and the anti-CD25 antibody to detect CD4+CD25+GPA33high T cells, and/or contacting the cell sample with an anti-CD127 antibody or antigen-binding fragment thereof and detecting binding or the lack thereof between cells in the sample and the anti-CD127 antibody to detect CD4+CD127−GPA33high T cells.
Relative threshold definition of GPA33high above the average on CD4+ T cells
Defining GPA33high as the expression level of GPA33 that is higher than the average level of expression of GPA33 on CD4+ T cells in the cell sample.
Proportional expression criterion for GPA33high based on 75% of a specified T-cell population
Defining GPA33high as an expression level at which 75% of CD4+CD25+CD45RA+ T cells are GPA33+.
Across the independent claims, the core claimed coverage centers on detecting CD4+ regulatory T-cell subsets using anti-CD4 and anti-GPA33 binding plus anti-CD25 and/or anti-CD127 binding or lack of binding, while GPA33high is defined either relative to the average GPA33 expression on CD4+ T cells in the sample or using a proportional criterion tied to 75% of CD4+CD25+CD45RA+ T cells being GPA33+.
Stated Advantages
Improved stability and suppressive function after expansion without rapamycin for adoptive immunotherapy using CD4+GPA33high Tregs.
Documented Applications
Diagnosing or classifying autoimmune disease associated with Treg insufficiency by quantifying CD4+GPA33high cells, optionally with CD25 patterns.
Cancer prognosis, typing, and therapy response prediction by comparing CD4+CD25+GPA33high T-cell levels in tumor samples versus reference samples, where higher tumor levels indicate poor prognosis or poor therapy response.
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