Details

Autor(en) / Beteiligte

• Yusubalieva, G. M.
• Dashinimaev, E. B.
• Gorchakov, A. A.
• Kulemzin, S. V.
• Brovkina, O. A.
• Kalinkin, A. A.
• Vinokurov, A. G.
• Shirmanova, M. V.
• Taranin, A. V.
• Baklaushev, V. P.

Titel

Enhanced Natural Killers with CISH and B2M Gene Knockouts Reveal Increased Cytotoxicity in Glioblastoma Primary Cultures

Ist Teil von

• Molecular biology (New York), 2022-10, Vol.56 (5), p.770-779

Ort / Verlag

Moscow: Pleiades Publishing

Erscheinungsjahr

2022

Quelle

Alma/SFX Local Collection

Beschreibungen/Notizen

• In an experimental study using the CRISPR/Cas9 system, “enhanced” NK cell lines with knockout of CISH , the gene for the CIS protein (a negative regulator of NK cytotoxicity), as well as two lines with a knocked-out β2-microglobulin gene, which provides membrane exposure of MHC class I, were obtained from two parental lines of human natural killers (YT wild type and YT-VAV1 + overexpressing the VAV1 cytotoxicity enhancing protein). The knockout efficiency was determined by real-time PCR as well as by flow cytometry with specific antibodies. The resulting CISH –/– or B2M –/– knockout lines were tested for cytotoxicity in primary monolayer cultures of human glioblastoma multiforme. The cytotoxicity of the lines was assessed using a cell analyzer that records the cell index based on cell impedance. YT-CISH –/– has been shown to be significantly more effective than wild-type YT in eliminating primary glioblastoma cells in an in vitro cell monolayer experiment. The cytotoxicity of the YT-VAV1 + -CISH –/– and YT-VAV1 + B2M –/– lines against glioblastoma cells was the highest, but overall, it did not significantly differ from the initially increased cytotoxicity of the YT-VAV1 + line. The lines of NK-like cells obtained may serve as a prototype for the creation of “enhanced” allogeneic and autologous NK- and CAR-NK cells for the immunotherapy of glioblastoma multiforme.