CRISPR/Cas9-mediated demethylation of FOXP3-TSDR toward Treg-characteristic programming of Jurkat T cells

2021 ◽  
pp. 104471
Author(s):  
Camilla Wilk ◽  
Laura Effenberg ◽  
Hanna Abberger ◽  
Laura Steenpass ◽  
Wiebke Hansen ◽  
...  
Keyword(s):  
T Cells ◽  
2009 ◽  
pp. 1-8
Author(s):  
Jing-Lei Qu ◽  
Xiu-Juan Qu ◽  
Ming-Fang Zhao ◽  
Yue-E Teng ◽  
Ye Zhang ◽  
...  

Materials ◽  
2021 ◽  
Vol 14 (13) ◽  
pp. 3693
Author(s):  
Yurii P. Sharkeev ◽  
Ekaterina G. Komarova ◽  
Valentina V. Chebodaeva ◽  
Mariya B. Sedelnikova ◽  
Aleksandr M. Zakharenko ◽  
...  

A modern trend in traumatology, orthopedics, and implantology is the development of materials and coatings with an amorphous–crystalline structure that exhibits excellent biocopatibility. The structure and physico–chemical and biological properties of calcium phosphate (CaP) coatings deposited on Ti plates using the micro-arc oxidation (MAO) method under different voltages (200, 250, and 300 V) were studied. Amorphous, nanocrystalline, and microcrystalline statesof CaHPO4 and β-Ca2P2O7were observed in the coatings using TEM and XRD. The increase in MAO voltage resulted in augmentation of the surface roughness Ra from 2.5 to 6.5 µm, mass from 10 to 25 mg, thickness from 50 to 105 µm, and Ca/P ratio from 0.3 to 0.6. The electrical potential (EP) of the CaP coatings changed from −456 to −535 mV, while the zeta potential (ZP) decreased from −53 to −40 mV following an increase in the values of the MAO voltage. Numerous correlations of physical and chemical indices of CaP coatings were estimated. A decrease in the ZP magnitudes of CaP coatings deposited at 200–250 V was strongly associated with elevated hTERT expression in tumor-derived Jurkat T cells preliminarily activated with anti-CD2/CD3/CD28 antibodies and then contacted in vitro with CaP-coated samples for 14 days. In turn, in vitro survival of CD4+ subsets was enhanced, with proinflammatory cytokine secretion of activated Jurkat T cells. Thus, the applied MAO voltage allowed the regulation of the physicochemical properties of amorphous–crystalline CaP-coatings on Ti substrates to a certain extent. This method may be used as a technological mechanism to trigger the behavior of cells through contact with micro-arc CaP coatings. The possible role of negative ZP and Ca2+ as effectors of the biological effects of amorphous–crystalline CaP coatings is discussed. Micro-arc CaP coatings should be carefully tested to determine their suitability for use in patients with chronic lymphoid malignancies.


Author(s):  
Kgomotso Welheminah Lebogo ◽  
Matlou Phineas Mokgotho ◽  
Victor Patrick Bagla ◽  
Thabe Moses Matsebatlela ◽  
Vusi Mbazima ◽  
...  

2012 ◽  
Vol 227 (6) ◽  
pp. 2556-2566 ◽  
Author(s):  
Chiung-Wen Tsao ◽  
Chiou-Feng Lin ◽  
Hung-Tsung Wu ◽  
Ching-Ting Ma ◽  
Wei-Ching Huang ◽  
...  

2007 ◽  
Vol 7 (2) ◽  
pp. 198-204 ◽  
Author(s):  
Radosław Januchowski ◽  
Paweł P. Jagodzinski

2021 ◽  
Vol 23 (Supplement_6) ◽  
pp. vi172-vi172
Author(s):  
Lei Ding ◽  
Lawrence Lamb

Abstract While recent advances in immunotherapies have shown promise in extracranial tumors, Glioblastoma Multiforme (GBM) has remained challenging. We have previously developed a novel approach to the treatment of primary GBM by combining simultaneous intracranial administration of gene-modified γδ T cells expressing O-6-Methylguanine-DNA Methyltransferase (MGMT) and standard temozolomide (TMZ) chemotherapy. To further improve tumor targeting and the efficacy of the γδ T cell against GBM, we modified γδ T cells with a CAR using chlorotoxin (CLTX), a small peptide derived from scorpion venom that specifically binds the glioma-specific chloride channel, as binding domains (CLTX-CARs). MGMTp140k was co-expressed within the same CLTX-CAR vectors to confer TMZ resistance to the CAR-T cells. We optimized the CAR constructs as CLTX-CD8a-CD28TM-CD28co-CD3z-P2A-MGMT and the activation of CD69 was verified in CLTX-CAR transduced Jurkat T cells with GBM cell co-culture. We further tested dual-CLTX-CAR constructs with 2x CLTX peptide as binding domains (dCLTX-CAR) and demonstrated dCLTX-CAR-Jurkat T cells can activate T cells efficiently with improved persistence compared to the mono-CLTX-CAR-Jurkat T cells 3 weeks after transduction (84% vs 37%). In order to preserve the powerful innate-recognition properties of γδ T cells, we developed CLTX-CARs without CD3z activation domain (NoZ-CLTX-CAR) to mitigate activation induced cell death (AICD) and tonic signaling issues. NoZ-CLTX-CAR-Jurkat T cells showed no CD69 activation with GBM cells co-culture and superior CAR-T persistence compared to CLTX-CARs with a CD3z signaling domain. Furthermore, we were able to efficiently transduce (up to 70% transduction efficiency) γδ T cells with NoZ-dCLTX-CAR lentiviral which demonstrated enhanced cytotoxicity against GBM cells compare to non-transduced control γδ T cells. Our approach of combining the dCLTX-CAR and TMZ resistance will be further validated in animal model experiments and could be a potential candidate for clinical development for GBM.


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