期刊
BIOMEDICINES
卷 10, 期 3, 页码 -出版社
MDPI
DOI: 10.3390/biomedicines10030630
关键词
SARS-CoV-2; T-lymphocytes; cytotoxicity; M protein
资金
- MutuaMadrilena Foundation [2020/0056]
- Plan Nacional de I+D+I
- Instituto de Salud Carlos III [COV20/00181]
- Spanish Ministry of Science and Innovation, Spanish Network for Research in Infectious Diseases [REIPI RD16/0016]
- European Development Regional Fund (EDRF)
- European Social Fund (ESF)
- REACT-EU grant from the Comunidad de Madrid to the ANTICIPA (Complutense University, Madrid)
- Sara Borrell Program [CD18CIII/00007]
- Instituto de Salud Carlos III, Ministerio de Ciencia, Innovacion y Universidades
- PFIS Program [F18III/00013]
The study demonstrates the ability to efficiently stimulate SARS-CoV-2-specific T-cells from convalescent donors and the increased percentage of these cells in vaccinated convalescent donors. These findings suggest the potential clinical usefulness of isolated SARS-CoV-2-specific T-cells.
In order to demonstrate the feasibility of preparing clinical-grade SARS-CoV-2-specific T-cells from convalescent donors and the ability of these cells to neutralize the virus in vitro, we used blood collected from two COVID-19 convalescent donors (before and after vaccination) that was stimulated with specific SARS-CoV-2 peptides followed by automated T-cell isolation using the CliniMacs Prodigy medical device. To determine cytotoxic activity, HEK 293T cells were transfected to express the SARS-CoV-2 M protein, mimicking SARS-CoV-2 infection. We were able to quickly and efficiently isolate SARS-CoV-2-specific T lymphocytes from both donors before and after they received the Pfizer-BioNTech vaccine. Althoughbefore vaccination, the final product contained up to 7.42% and 30.19% of IFN-gamma+ CD3+ T-cells from donor 1 and donor 2, respectively, we observed an enrichment of the IFN-gamma+ CD3+ T-cells after vaccination, reaching 70.47% and 42.59%, respectively. At pre-vaccination, the isolated SARS-CoV-2-specific T-cells exhibited cytotoxic activity that was significantly higher than that of unstimulated controls (donor 2: 15.41%, p-value 3.27 x 10(-3)). The cytotoxic activity of the isolated SARS-CoV-2-specific T-cells also significantly increased after vaccination (donor 1: 32.71%, p-value 1.44 x 10(-5); donor 2: 33.38%, p-value 3.13 x 10(-6)). In conclusion, we demonstrated that SARS-CoV-2-specific T-cells can quickly and efficiently be stimulated from the blood of convalescent donors using SARS-CoV-2-specific peptides followed by automated isolation. Vaccinated convalescent donors have a higher percentage of SARS-CoV-2-specific T-cells and may be more suitable as donors. Although further studies are needed to assess the clinical utility of the functional isolated SARS-CoV-2-specific T-cells in patients, previous studies using the same stimulation and isolation methods applied to other pathologies support this idea.
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