A highly compact epitope based marker suicide gene for easier ashpublications org blood article 124 8 1277 33527 A highly compact epitope based marker suicide gene
Springer article 10 Functional Validation of the RQR8 Suicide Marker Gene in Successful Construction of Rqr8 CAR T Cells Rqr8 Tag Did Not Affect The Phenotype of CAR T Cells Rituximab Induced ADCC Can Eliminate Rqr8 CAR T Cells Rituximab Induced CDC Can Eliminate Rqr8 CD19 CAR T Cells Rqr8 CD19 CAR T Cells Can Be Eliminated by Rituximab in Vivo A CAR sequence was composed of a scFv derived from FMC63 CD137 41BB co stimulatory and CD3zeta signaling domains and then combined with the RQR8 sequence linked by the T2A peptide The whole sequence was cloned into a lentiviral expression vector and a myeloproliferative sarcoma virus enhancer dl587Rev primer binding site substituted MND promoter was used to ensure the stable expression of RQR8 T2A CAR sequences in T cells Fig 1a We also constructed a CAR vector Fig 1b targeting CLL1 which is highly expressed on leukemia stem cells and acute myeloid leukemia primitive cells We then tested the CAR expression of prepared CAR T cells with the QBEND 10 monoclonal CD34 specific antibody Fig 1c d To confirm the consistency of CAR and RQR8 expression we also stained CD19 CAR T cells with an anti FMC63 antibody and before staining CLL1 CAR T cells we incubated the CLL1 protein carrying a HIS tag with an anti HIS antibody The results showed that there was no signific See full list on link springer com Previous studies have reported that CAR T cells with a high proportion of naïve T cells CD62L CD45RO have a better therapeutic effect than those with a low proportion while the proportion of naïve T cells in virus transduced T cells is significantly lower than that in nontransduced T cells 22 23 24 To explore the influence of the RQR8 tags on the CAR T cell phenotypes we detected the proportion of naïve T cells on UCAR T cells and RQR8 CAR T cells by flow cytometry The results showed that there were no significant differences in T cell phenotype between the UCAR T cells and RQR8 CAR T cells Fig 2a b Subsequently we detected the apoptosis of nontransduced T cells UCAR T cells and RQR8 CAR T cells on days 4 7 and 12 of cell culture Fig 2c d These results suggested that the influence on the phenotype effect of CAR T cells was negligible and that the addition of the RQR8 tag did not increase CAR T cell apoptosis See full list on link springer com NK cells from different sources also kill T cells Thus we used peripheral blood from the same healthy volunteers to prepare NK cells and CAR T cells On the 14th day of expansion we obtained highly purified NK cells 97 75 using a commercial NK cell amplification kit Fig 4a We coincubated homologous NK cells and RQR8 CAR T cells at an E T ratio of 1 1 with or without 50 μg ml rituximab Fig 4b which is a concentration below the effective clinical dose Twenty four hours later the numbers of live RQR8 CAR T cells were detected by flow cytometry The results showed that the number of RQR8 CAR T cells was significantly reduced in the rituximab group Fig 4c d To further verify our conclusions we incubated NK cells RQR8 CD19 CAR T cells and tumor cells at an equal ratio and used coincubation controls of NK cells T cells and tumor cells with or without rituximab 50 μg ml After 24 h of incubation we used flow cytometry to detect the percentage of remaining tumor See full list on link springer com We further validated the depletion of RQR8 CAR T cells caused by the rituximab induced CDC effect The classical complement pathway was activated when rituximab was combined with the CD20 epitopes of RQR8 CAR T cells and ultimately eliminated the RQR8 CAR T cells Fig 5a We added 12 baby rabbit complement to RQR8 CAR T cells with or without rituximab Twelve hours later the survival rate of RQR8 CAR T cells was detected and analyzed Fig 5b We performed the RQR8 CD19 CAR T cell coincubations in the presence of 12 rabbit complement with or without rituximab Any remaining RQR8 CD19 CAR T cells were detected at 30 min 1 h 2 h 4 h 6 h and 12 h The results showed that a large number of RQR8 CD19 CAR T cells were lost after the addition of rituximab and the loss trend became increasingly obvious over time with almost no RQR8 CAR T cells detected at 12 h Fig 5c To further verify the accuracy of this conclusion RQR8 CAR T cells were incubated with tumor cells at an See full list on link springer com We transplanted Nalm 6 cells expressing luciferase and green fluorescent protein into irradiated immunodeficient mice and injected the mice with T cells or RQR8 CD19 CAR T cells on day 3 after tumor cell injection In vivo imaging was performed to observe tumor growth on day 5 after injection and the RQR8 CAR T cell group was divided into a rituximab positive group and a rituximab negative group according to whether rituximab was injected Fig 6a b The results showed that RQR8 CD19 CAR T cells had an antitumor effect in vivo and extended the survival time of mice compared to nontransduced T cells In addition we observed that the antitumor effect of the CAR T cells was reduced when CAR T cells were eliminated followed by the injection of rituximab Fig 6c d See full list on link springer com Author Xia Xiong
Nature articles s41434 021 00220 6 Targeted multi epitope switching enables straightforward Feb 1 2021 The epitope spectrum of esCAR T cells provided by RQR8 facilitates enrichment of gene edited T cells using commercially available selection systems
PubMed 24970931 A highly compact epitope based marker suicide gene for easier Aug 21 2014 We have created a highly compact marker suicide gene for T cells combining target epitopes from both CD34 and CD20 antigens RQR8 This construct allows selection with
UCL Discovery 1470296 RQR8 A universal safety switch for cellular therapies We sought to generate a compact marker suicide gene that enables clinical grade sorting and effective deletion using CD34 cliniMACS and rituximab respectively By using Author BM Philip Publish Year 2015 Event University College London Type Thesis Doctoral
ResearchGate publication 363543762 Combinatorial Combinatorial suicide gene strategies for the safety of cell combining the DiC9 with the RQR8 compact suicide gene RQR8 incorporates a CD20 mimotope targeted by the anti CD20 monoclonal antibody rituxan and the QBend10 a DCD34 People also search for
Rqr8 Gene
pmc ncbi nlm nih gov articles PMC9515659 Combinatorial suicide gene strategies for the safety of cell A second approach was employed in parallel combining the ΔiC9 with the RQR8 compact suicide gene RQR8 incorporates a CD20 mimotope targeted by the anti CD20 monoclonal
Refine this search rqr8 gene therapy rqr8 gene mutation rqr8 gene editing rqr8 gene testing rqr8 gene transfer rqr8 gene expression
Rqr8 Gene
Image researchgate net 136 amino acid epitope based marker suicide gene RQR8 is a 136 amino acid epitope based marker suicide gene that enables clinical selection cell tracking and deletion in case of toxicity
Cell Press molecular therapy family molecular therapy 86 A Highly Compact Epitope Based Marker Suicide Gene for We demonstrated almost complete in vivo depletion of RQR8 expressing T cells within 7 days of antibody administration and subsequent resolution of GvHD Conclusions RQR8
Springer content pdf Functional Validation of the RQR8 Suicide Marker Gene in RQR8 is a gene that integrates CD34 and CD20 epitopes In our study we incorporated the suicide gene RQR8 into CAR T cells so it enabled rituximab to eliminate
PubMed 36189285 Combinatorial suicide gene strategies for the safety of cell Sep 14 2022 RQR8 incorporates a CD20 mimotope targeted by the anti CD20 monoclonal antibody rituxan and the QBend10 a ΔCD34 selectable marker Likewise enhanced cell elimination