CD4+ T Cell Marker Enrichment for Tumor-Specific TIL Retention
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
The phenotypic states and functional roles of tumor antigen-specific CD4+ T cells in human cancers are poorly understood, and these cells are often lost during conventional tumor infiltrating lymphocyte (TIL) culture, hindering their therapeutic and prognostic applications.
Innovation Solution
Identify CD4+ T cells in solid tumors by specific markers such as CXCL13 expression, and use CXCL13+CD200+ or CXCL13+PD-1+CD200+ as surrogates to isolate and enrich for tumor antigen-specific CD4+ T cells, which can be expanded and used in T cell therapy, and their TCRs can be sequenced for therapeutic applications.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If conventional TIL culture methods are used, then general T cell expansion is achieved, but tumor antigen-specific CD4+ T cells are lost
Solution Approach 1:
The patent extracts and isolates tumor antigen-specific CD4+ T cells from the heterogeneous TIL population using specific phenotypic markers (CXCL13+, PD-1+, CD200+) before expansion. This extraction step separates the desired cells from the bulk population that would be lost in conventional culture, enabling selective enrichment and expansion of the specific subset.
Solution Approach 2:
The patent uses phenotypic markers (CXCL13, PD-1, CD200) as intermediaries to identify and select tumor antigen-specific CD4+ T cells. These markers serve as mediators that enable the separation and enrichment of the specific cell subset without requiring prior knowledge of antigen specificity, solving the problem of losing these cells in conventional culture.
2Measurement precision
If phenotypic markers are used to identify tumor antigen-specific CD4+ T cells, then detection accuracy is improved, but measurement complexity increases
Solution Approach 1:
The patent employs a universal phenotypic marker profile (CXCL13+, PD-1+, CD200+) that can identify tumor antigen-specific CD4+ T cells across multiple cancer types and antigen specificities. This multi-functional marker set eliminates the need for antigen-specific assays for each individual cell, simplifying the overall identification process while maintaining high precision.
Solution Approach 2:
The patent changes the detection parameters from functional assays (which measure antigen-specific activity) to phenotypic marker expression (which measures surface protein levels). This parameter change enables high-throughput flow cytometry-based identification, improving detection accuracy while managing complexity through standardized marker panels.
3Reliability
If tumor antigen-specific CD4+ T cells are enriched and expanded, then therapeutic efficacy is improved, but production time increases
Solution Approach 1:
The patent performs preliminary enrichment of tumor antigen-specific CD4+ T cells using phenotypic markers before expansion. By pre-selecting the specific cell subset beforehand, the subsequent expansion phase can proceed more efficiently with shorter timescales, as only the relevant cells are being cultured rather than attempting to expand the entire TIL population.
Solution Approach 2:
The patent segments the TIL processing into distinct phases: (1) phenotypic enrichment of CD4+ T cells, (2) separation from CD8+ T cells, and (3) targeted expansion. This segmentation allows each phase to be optimized independently, reducing overall production time while maintaining therapeutic efficacy through focused expansion of the specific cell subset.
Data Source
AI summary
The present disclosure relates, in part, to the discovery that CD4+ T cells characterized by certain markers are over-represented in solid tumors. Markers include, for example, CD200, CXCL13, and PD-1, as well as combinations of these, and other markers. The markers can identify CD4+ T cells having certain phenotypes. Identifying such cells enables a variety of therapeutic and prognostic applications. Such CD4+ T cells will typically include those specific for a tumor neoantigen or antigen, and may be enriched for and/or expanded and used in a T cell therapy. TCRs from such CD4+ T cells may be used in therapy (e.g., comprising T cells recombinantly expressing such a TCR or an engineered TCR or binding domain derived therefrom). The relative presence or absence of such CD4+ T cells in a tumor sample can provide information regarding the immune state of the subject and the likely prognosis of disease.