Generation of an artificial human B cell line test system using Transpo-mAbTM technology to evaluate the therapeutic efficacy of novel antigen-specific fusion proteins.
Diana KloseMira WoitokJudith NiesenRoger R BeerliUlf GrawunderRainer FischerStefan BarthRolf FendelThomas NachreinerPublished in: PloS one (2017)
The antigen-specific targeting of autoreactive B cells via their unique B cell receptors (BCRs) is a novel and promising alternative to the systemic suppression of humoral immunity. We generated and characterized cytolytic fusion proteins based on an existing immunotoxin comprising tetanus toxoid fragment C (TTC) as the targeting component and the modified Pseudomonas aeruginosa exotoxin A (ETA') as the cytotoxic component. The immunotoxin was reconfigured to replace ETA' with either the granzyme B mutant R201K or MAPTau as human effector domains. The novel cytolytic fusion proteins were characterized with a recombinant human lymphocytic cell line developed using Transpo-mAb™ technology. Genes encoding a chimeric TTC-reactive immunoglobulin G were successfully integrated into the genome of the precursor B cell line REH so that the cells could present TTC-reactive BCRs on their surface. These cells were used to investigate the specific cytotoxicity of GrB(R201K)-TTC and TTC-MAPTau, revealing that the serpin proteinase inhibitor 9-resistant granzyme B R201K mutant induced apoptosis specifically in the lymphocytic cell line. Our data confirm that antigen-based fusion proteins containing granzyme B (R201K) are suitable candidates for the depletion of autoreactive B cells.
Keyphrases
- induced apoptosis
- endoplasmic reticulum stress
- signaling pathway
- oxidative stress
- endothelial cells
- pseudomonas aeruginosa
- recombinant human
- immune response
- cell cycle arrest
- cancer therapy
- genome wide
- induced pluripotent stem cells
- cell therapy
- cystic fibrosis
- dna methylation
- regulatory t cells
- gene expression
- electronic health record
- machine learning
- cell death
- big data
- cell proliferation
- dendritic cells
- artificial intelligence
- acinetobacter baumannii
- data analysis
- candida albicans