Double Negative T Cell Expansion via Mitogen Depletion
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Solution Overview
Problem
Current cancer immunotherapy methods, such as adoptive transfer of CD8+ cytotoxic T lymphocytes, face limitations including short survival of infused cells, risk of graft-versus-host disease, and difficulty in identifying relevant tumor antigens, which restrict their effectiveness and safety.
Innovation Solution
A method for expanding double negative (DN) T cells ex vivo by purifying them from peripheral blood, depleting CD8+ and CD4+ T cells, and culturing with immobilized T cell mitogens and cytokines, allowing for their potent anti-tumor effects without causing graft-versus-host disease.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If CD8+ cytotoxic T lymphocytes are used for adoptive cancer immunotherapy, then anti-tumor response is achieved, but graft-versus-host disease occurs and cell survival is short
Solution Approach 1:
The patent extracts and isolates a specific subset of T cells (CD4-CD8- double negative T cells) from the broader T cell population. These cells are purified through flow cytometry-based separation, removing conventional CD4+ and CD8+ T cells that cause GVHD, while retaining the anti-tumor active DN T cells. This extraction resolves the contradiction by eliminating the harmful GVHD component while preserving the beneficial anti-tumor response.
Solution Approach 2:
The patent applies local quality by identifying and utilizing a specific subset of T cells with unique characteristics (CD4-CD8- phenotype) that have different functional properties from conventional T cells. These double negative T cells exhibit localized functionality - they provide anti-tumor activity without the harmful GVHD effect of conventional T cells, thus resolving the contradiction through selective targeting of a specific cell population with specialized properties.
2Reliability
If CD8+ cytotoxic T lymphocytes are expanded ex vivo, then anti-tumor activity is enhanced, but cell survival duration is limited to a few days
Solution Approach 1:
The patent changes the fundamental parameter of T cell subset selection from conventional CD8+ cells to CD4-CD8- double negative T cells. This parameter change results in cells with fundamentally different survival characteristics - the DN T cells demonstrate prolonged survival and persistence in vivo compared to the short-lived CD8+ cells, thereby resolving the contradiction between enhanced anti-tumor activity and limited survival duration.
3Reliability
If tumor specific CD8+ CTLs are identified and expanded, then effective anti-tumor response is achieved, but identification of relevant tumor antigens becomes complex
Solution Approach 1:
The patent applies universality by using double negative T cells that can recognize and respond to multiple types of tumor antigens through their unique T cell receptor characteristics. These cells exhibit broad reactivity against various tumor types without requiring identification of specific tumor antigens, thereby resolving the contradiction between achieving effective anti-tumor response and the complexity of antigen identification.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
Ex vivo expanded DN T cells demonstrate greater anti-tumor efficacy and longer survival compared to CD8+ T cells, both in vitro and in vivo, without inducing graft-versus-host disease, making them a promising novel approach for cancer immunotherapy.
Implementation Method 1
culturing the sample from step (b) with an immobilized T cell mitogen in a culture medium comprising an agent that can stimulate the growth of DN T cells
Implementation Method 2
washing the cells obtained in step (d) and resuspending in a culture medium comprising the agent and a soluble T cell mitogen
Data Source
AI summary
A method of expanding double negative T cells in culture is described. The method comprises (a) providing a starting sample comprising DN T cells or precursors thereof; (b) substantially depleting CD8+ and CD4+ T cells from the starting sample; (c) culturing the sample from step (b) with an immobilized T cell mitogen in a culture medium comprising an agent that can stimulate DN T cell growth (d) washing the cells obtained in step (c) and resuspending in a culture medium comprising the agent without the T cell mitogen; and (e) washing the cells obtained in step (d) and resuspending in a culture medium comprising the agent and a soluble T cell mitogen. The DN T cells obtained by the method are useful in a variety of applications including the treatment of cancer, infectious diseases, graft versus host disease and autoimmune disease.