Journal: bioRxiv

Article Title: Development of a recombinant adeno-associated virus vector for human T lymphocyte- and natural killer cell-targeted gene therapy

doi: 10.64898/2026.02.26.707014

Figure Lengend Snippet: Transduction of SupT1 (A) and Jurkat (B) cells with wild-type (wt) and CD7-engineered AAVs of different serotypes (10,000 gc/cell). Shown are relative % GFP-positive cells at 3 dpi compared to corresponding wt AAV serotype (mean ± SD, n=3). Absolute percentage GFP-positive cells are indicated inside the bars. C ) Representative flow cytometry plots of activated human PBMCs transduced with indicated rAAVs (100,000 gc/cell). D) Absolute % GFP-positivity at 3 dpi (mean ± SD, n≥4). Individual values are presented as dots. E) CD7-specificity factor for transduction (10,000 gc/cell) of activated PBMCs at 3 dpi (mean ± SD, n=3). F) Transduction of activated human PBMCs with indicated rAAVs (100,000 gc/cell) and stratification of GFP-positive cells at 3 dpi in CD4 + T, CD8 + T and NK (CD56 + ) cell types (mean ± SD, n≥3; dot colors represent biologically independent human cell samples). G) Transduction of hepatocyte (HuH-7, HepG2) and kidney (HEK293T) cell lines with indicated rAAVs (100.000 gc/cell). Absolute % GFP-positive cells at 3 dpi (mean ± SD, n=3). rAAV Vectors contained the following sequences: AAV6 & AAV2 = self-complementary genome, CAG promoter; AAV9 & AAV6/9 = single stranded genome, SFFV promoter. Mean ± SD, n=3. For all panels statistical analysis utilized unpaired t-test, p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), and p < 0.0001 (****).

Article Snippet: Suspension cell lines SupT1 (ATCC: CRL-1942) and Jurkat (RRID: CVCL_C831) were maintained in RPMI 1640 (Biozym Scientific) with 10% FCS, 50 U/mL Penicillin and 50 μg/mL Streptomycin (Pan Biotech).

Techniques: Transduction, Flow Cytometry

Journal: bioRxiv

Article Title: The central pore of HIV-1 capsomers promotes sustained stability of the viral capsid

doi: 10.1101/2025.05.19.654868

Figure Lengend Snippet: (A-C) Early RT products (A), late RT products (B), and 2-LTR circles (C) were quantified for WT, SK, LSK, SKT, SKI, and NSK at 1, 2, 3, 6, 9, and 12 hours post-infection (hpi) of SupT1 cells by qPCR. Each individual sample was quantified relative to CCR5 copy number to control for the cell number, and all data for each viral DNA species were normalized to copy number/CCR5 for WT at 12 hpi. Error bars depict the mean ± s.e.m. from 3 independent experiments. (D-F) minus-strand strong stop (D, MSSS), first-strand transfer (E, FST), and second-strand transfer (F, SST) DNA copies were quantified by qPCR in endogenous reverse transcription (ERT) reactions performed for 6 hours at 37°C with Streptolysin O (SLO) and in the presence or absence of DNase. Error bars depict the mean ± s.e.m. from at least 2 independent experiments. (G-H) Comparison of mean LSK and NSK early (G) and late (H) RT products at 6 hours post infection in SupT1 cells relative to WT to MSSS (G) and SST (H) RT products in ERT reactions performed in the presence and absence of DNase. Abbreviations: R18S/N21K (SK); S16L/R18S/N21K (LSK); R18S/N21K/A31T (SKT); R18S/N21K/T216I (SKI); S16N/R18S/N21K (NSK).

Article Snippet: MT4 and SupT1 T-cell lines were acquired from American Type Culture Collection (ATTC) and cultured in Roswell Park Memorial Institute (RPMI) 1640 medium supplemented with 10% fetal bovine serum, 2mM L-glutamine, penicillin (100U/mL), and streptomycin (100 μg/mL).

Techniques: Infection, Control, Reverse Transcription, Comparison

Journal: Nature Communications

Article Title: HIV-1 adapts to lost IP6 coordination through second-site mutations that restore conical capsid assembly

doi: 10.1038/s41467-024-51971-w

Figure Lengend Snippet: A, B MT4 cells were transfected with infectious molecular clones harboring mutations at either R18 ( A ) or K25 ( B ) and replication kinetics were assessed by quantifying supernatant RT activity. C Supernatants from ( B ) and similar repeat experiments were used to infect MT4, C8166, or SupT1 T cell lines to assess replication kinetics after acquisition of compensatory mutations. Infectious molecular clones of N21S ( D ), T216I ( E ), G208R ( F ), A105T ( G ) and G225S ( H ) variants were used to transfect MT4 cells to assess replication kinetics.

Article Snippet: MT4, C8166, and SupT1 human T cell lines were acquired from American Type Culture Collection (ATCC) and cultured in RPMI 1640 medium (Corning) supplemented with 10% fetal bovine serum (GenClone), 2 mM L-glutamine, penicillin (100U/ml; Gibco), and streptomycin (100μg/mL; Gibco).

Techniques: Transfection, Clone Assay, Activity Assay