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. 2023 Mar 14;11(3):e006292. doi: 10.1136/jitc-2022-006292

Figure 6.

Figure 6

Biodistribution of surface-engineered lentiviral vectors in a canine. (A) Canines were treated with either 4E+08 TU CD3-cocal-GFP via ultrasound-guided bilateral inguinal lymph node injection or 4E+9 TU CD3-cocal-GFP via intraperitoneal injection. Necropsy was performed after either 1 or 4 weeks to assess lentiviral integration biodistribution profiles. (B) Biodistribution of transduced cells was evaluated by detection of the viral element PSI in genomic DNA blood and tissue samples using qPCR. Only organs with transduction events detected over the LLOQ are shown. (C) RNA ISH was performed to characterize the identity of cells expressing EGFP RNA transcripts using colocalization analysis. Representative images depict canine T cells transduced by CD3-cocal-GFP in the inguinal lymph node, medial iliac lymph node, and spleen 1 week after intranodal injection. Original magnification ×40; scale bars indicate 5 µM. White denotes DAPI nuclear stain; green denotes eGFP of transduced cells; yellow denotes canine CD3; red denotes canine CD68; blue denotes canine CD45. eGFP, enhanced green fluorescent; ISH, in situ hybridization; IP, intraperitoneal; LLOQ, lower limit of quantification; LN, lymph node;TU, transducing unit.