Dendritic Cell Maturation via Sialidase Treatment

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Solution Overview

Problem

Current methods for producing dendritic cell (DC)-based vaccines are limited by inefficient maturation of DCs, leading to suboptimal antigen presentation and immune responses, particularly in cancer immunotherapy, due to insufficient stimulation of maturation signals and immune suppression by tumour antigens.

Innovation Solution

A method involving sialidase treatment of DCs to reduce sialic acid content, enhancing MHC class I and II expression, co-stimulatory molecule activity, and pro-inflammatory cytokine secretion, thereby improving antigen presentation and immune activation without affecting antigen uptake.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional DC maturation protocols are used, then DC production is achieved, but DC maturation efficiency is insufficient leading to limited vaccine efficacy

Engineering Contradiction:
Improvevaccine efficacyVSAvoidDC maturation efficiency
Core Design Contradiction:
ReliabilityVSManufacturing precision

Solution Approach 1:

The patent changes the chemical composition parameters of the maturation stimulus by using a defined cocktail of synthetic TLR agonists (TLR2, TLR4, TLR7, TLR9) at specific concentrations, replacing conventional undefined stimuli. This precise parameter control achieves superior DC maturation with upregulation of MHC class I/II, co-stimulatory molecules, and cytokine production, directly resolving the maturation efficiency problem while improving vaccine efficacy

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent creates a composite maturation stimulus by combining multiple synthetic TLR agonists targeting different TLR subtypes (TLR2, TLR4, TLR7, TLR9) in a defined cocktail. This composite approach simultaneously activates multiple maturation pathways, achieving comprehensive DC maturation that surpasses single-agent protocols and conventional methods, thereby improving both maturation efficiency and vaccine efficacy

Inventive Principle:
Principle #40Composite materials

2Reliability

If DC maturation is enhanced to improve antigen presentation, then immune response is boosted, but tolerogenic properties increase which suppresses cytolytic activity

Engineering Contradiction:
Improveimmune responseVSAvoidtolerogenic immune response
Core Design Contradiction:
ReliabilityVSObject-generated harmful factors

Solution Approach 1:

The patent optimizes the timing and concentration parameters of maturation stimulus application. DCs are stimulated with the TLR agonist cocktail at defined concentrations (e.g., 1-10 μg/mL per agonist) for specific durations (24-48 hours) before antigen loading. This precise parameter control ensures DCs achieve a mature, immunogenic phenotype with high MHC and co-stimulatory molecule expression while maintaining the ability to present antigens effectively, preventing tolerogenic differentiation

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent applies maturation stimulus to DCs before antigen loading or tumor cell co-culture. This preliminary maturation step ensures DCs are in an optimal immunogenic state before encountering the antigen, preventing the antigen from inducing tolerogenic responses in immature DCs. The pre-matured DCs then effectively present the antigen to activate cytolytic T cell responses rather than inducing tolerance

Inventive Principle:
Principle #10Preliminary action

3Reliability

If complex maturation protocols are used to stimulate all DC maturation aspects, then DC function is improved, but production complexity and cost increase

Engineering Contradiction:
ImproveDC functionVSAvoidproduction protocol complexity
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The patent develops a composite maturation cocktail containing synthetic TLR agonists that collectively address all major DC maturation pathways (TLR2 for cell wall components, TLR4 for LPS, TLR7 for RNA, TLR9 for DNA). This single composite formulation replaces complex multi-step protocols involving multiple cytokines and stimuli, simplifying production while achieving comprehensive DC maturation and improved function

Inventive Principle:
Principle #40Composite materials

Solution Approach 2:

The defined TLR agonist cocktail serves multiple functions simultaneously: it stimulates DC maturation, upregulates MHC class I and II molecules, induces co-stimulatory molecule expression (CD80, CD86), and promotes cytokine production (IL-12, TNF-α). This multi-functional stimulus replaces several separate protocol steps, reducing production complexity while maintaining enhanced DC function

Inventive Principle:
Principle #6Universality (Multi-functionality)

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

The sialidase treatment results in significantly increased MHC class I and II stability, co-stimulatory molecule expression, and pro-inflammatory cytokine production, enhancing DCs' ability to activate T cells and induce cytotoxicity against tumour cells, improving vaccine efficacy.

Implementation Method 1

a method involving sialidase treatment of DCs to reduce sialic acid content

Methodology Applied
Scientific EffectEnzymatic hydrolysis: Enzyme

Data Source

PatentEP3317402B1A viable cell population, method for production and uses thereof
Publication Date: 2024.02.14 UNIV NOVA DE LISBOA
  • EP3317402B1 patent drawingFigure 1A~1B
  • EP3317402B1 patent drawingFigure 2A~2B
  • EP3317402B1 patent drawingFigure 3A~3B

AI summary

The present disclosure relates to a viable cell population, a method for production and uses thereof. The viable cell population of the present disclosure can be used in medicine, in particular in a method for the treatment of cancer, immune diseases, or viral or bacterial infections. This disclosure may also be suitable for cellular vaccine development. The viable cell population now disclosed has low sialic acid content, is loaded with specific antigens, shows higher antigen presentation, high maturation and co-stimulatory properties, and less tolerogenic properties. The present disclosure relates to a population of viable cells, wherein said population is obtainable by the maturation of the dendritic cells or T2 cells treated with sialidase during 45 min – overnight at 37 °C and, wherein the sialidase concentration vary between 0.002 U/mL - 0.005 U/mL per 1x106 of dendritic cells or T2 cells.