Virus-Specific T Cell Profiling Using Peptide Pools
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Solution Overview
Problem
Current methods for quantifying virus-specific T cells are complex and cumbersome, making it difficult to measure cellular immunity effectively, particularly in the context of viral infections like COVID-19, where antibody detection may not suffice to gauge antiviral immunity.
Innovation Solution
A method involving the use of peptide pools derived from virus antigenic proteins to stimulate T cells, allowing for the discrimination between infected and uninfected individuals, assessment of vaccine efficacy, and evaluation of the impact of viral variants on T cell responses through ELISPOT, qPCR, and other techniques.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional methods (ELISPOT, intracellular cytokine staining, qPCR) are used to quantify virus-specific T cells, then measurement precision is improved, but device complexity and ease of operation deteriorate due to requiring specialized skills, equipment, and complex procedures including PBMC separation
Solution Approach 1:
The patent segments the complex T cell detection process into a simplified format by using pre-separated whole blood samples and standardized peptide pools, dividing the complexity from the user while maintaining detection accuracy through structured assay protocols
Solution Approach 2:
The patent introduces an intermediary standardized assay format that mediates between complex research-grade T cell detection methods and simple clinical applications, using pre-prepared reagents and standardized protocols to bridge the gap between precision and ease of use
2Measurement precision
If conventional methods are used to quantify virus-specific T cells, then measurement precision is improved, but ease of operation worsens due to requiring specialized technical personnel and complex experimental procedures
Solution Approach 1:
The patent performs preliminary actions by pre-separating and preparing whole blood samples, pre-formulating peptide pools, and establishing standardized protocols before the actual T cell detection, thereby eliminating complex steps from the user's workflow while maintaining measurement precision
Solution Approach 2:
The patent employs disposable pre-prepared reagents and standardized assay kits that can be used once and discarded, eliminating the need for specialized equipment and technical expertise while maintaining detection accuracy through manufacturer-optimized protocols
3Ease of operation
If antibody detection methods are used to gauge antiviral immunity, then ease of operation is improved, but measurement precision worsens because antibody titers may not fully reflect cellular immunity levels
Solution Approach 1:
The patent creates a universal assay platform that can detect both humoral (antibody) and cellular (T cell) immunity responses using the same basic workflow and sample type, allowing comprehensive immunity assessment without requiring separate specialized tests for each immune component
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
Enables accurate discrimination between infected and uninfected individuals, evaluates vaccine-induced T cell responses, and quantifies the impact of viral variants on T cell immunity, providing a more comprehensive understanding of antiviral immunity beyond antibody detection.
Implementation Method 1
assaying a sample comprising or derived from blood, broncholavage (BAL fluid), nasal swabs, or nasopharyngeal aspirate from a subject to determine whether it comprises T cells reactive to one or more virus peptide pools, wherein said peptide pools are separately derived from virus antigenic structural and/or non-structural proteins
Implementation Method 2
The profiling is typically performed using ELISPOT, but may also be performed using other techniques such as qPCR, more particularly direct qPCR
Data Source
AI summary
The present invention relates to a method of diagnosing and/or monitoring of virus infection and/or response to vaccination by generating a profile of a virus-specific T cell response that can (i) discriminate between virus infected and uninfected individuals, (ii) determine the effect of vaccination on T cell response, and (iii) determine the effect of viral variants on T cell response. More particularly, the described virus-specific T cell profiling is based on the detection of activated antigen-specific T lymphocytes responding to pools of selected short peptides from virus proteins. These peptide sequences have been selected for their immunogenicity. The profiling is typically performed using ELISPOT, but may also be performed using other techniques such as qPCR, more particularly direct qPCR. The present invention also includes kits for use in the methods of the invention.