B12A-4G4，A11-4G4 NTCC® TCR-negative 4G4 T cell line小鼠杂交瘤细胞株-BioVector NTCC Inc.

B12A-4G4，A11-4G4 NTCC® TCR-negative 4G4 T cell line小鼠杂交瘤细胞株-BioVector NTCC Inc.

Full-length TCR cDNA for TCRα and TCRβ were cloned with PCR primers specific to sequences 5′ of the start and 3′ of the stop codon of rearranged TCR chains. TCR expression constructs were cloned into murine stem cell virus–based retroviral plasmids with pMX-IRES-GFP (provided by R. Bosselut, National Cancer Institute) for TCRβ and MSCV-IRES-tNGFR (provided by W. Pear, University of Pennsylvania) for TCRα, and they were transfected into PlatE cells separately to produce retrovirus containing supernatants. TCR-negative 4G4 T cells were retrovirally transduced with both TCRα- and TCRβ-containing retroviruses in the presence of 2 μg/ml of polybrene. The resulting B12A-4G4 or A11-4G4 cells (5 × 10⁴ cells per well) were incubated overnight at 37°C with B6 or MHC^KO stimulator spleen cells that had been treated overnight with LPS (10 μg/ml), irradiated with 3000R, and T depleted. Purified B10.A lymph node T cells that were alloreactive against H-2^b stimulators were used as control responder T cells. MHC blocking Abs used were a mixture of mAbs specific for MHC class I anti–H-2K^b/H-2D^b (28-8-6) and MHC class II Y3P (I-A), M5/114 (I-A/I-E) at 10 μg/ml each. Culture supernatants of stimulated B12A- and A11-4G4 cells were collected, and IL-2 concentrations were measured by ELISA (R&D Systems).

The 4G4 cell line is a mouse hybridoma cell line that produces a monoclonal antibody against the influenza D virus hemagglutinin protein. This antibody is used in research to study the influenza D virus and to develop antiviral drugs.

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