Enhanced transduction of antigen-stimulated T lymphocytes with recombinant retroviruses concentrated by centrifugal filtration

T‐helper type 2‐dependent early recruitment of antigen non‐specific CD4+ T cells in experimental asthma

“…1a). As we have shown previously [8, 25], when total splenocytes are cultured with antigen, CD4 + T cells are preferentially transduced (Fig. 1b).…”

“…Following culture with antigen, the T cells that originally encountered antigen in vivo during sensitization become reactivated and undergo proliferation, allowing them to be transduced with recombinant retroviruses [25,26]. As we and others have shown previously, a subset of the transduced cells are OVAspecific [8,25,26]. Consistent with this, upon sequential in vitro exposure to OVA and recombinant retroviruses, CD4 1 T cells are selectively transduced ( Fig.…”

“…Moreover, these CD4 1 T cell-dependent allergen-driven inflammatory responses in the lung and draining lymph nodes were augmented in recipients of CD4 1 T cells overexpressing the Th2-specific GATA-3 transcription factor. By stimulating splenocytes from OVA-sensitized BN rat donors with antigen in vitro, we are able to obtain a population of T cells that can be efficiently retrovirally transduced in numbers sufficient for adoptive transfer experiments [8,25]. Following culture with antigen, the T cells that originally encountered antigen in vivo during sensitization become reactivated and undergo proliferation, allowing them to be transduced with recombinant retroviruses [25,26].…”

“…Retroviruses were harvested from Phoenix‐Eco packaging cells stably transfected with the pAP2 retroviral vector, which encodes enhanced GFP (EGFP) only [24], or its derivative containing the full‐length GATA‐3 cDNA isolated from rat TIB‐236 cells and cloned upstream of the IRES‐EGFP cassette in pAP2. Recombinant retroviruses, concentrated 10‐fold by centrifugal filtration, had titres on NIH3T3 fibroblasts in the range of 5–10 × 10 6 CFU/mL [25]. Stimulated splenocytes were transduced with the control retrovector encoding EGFP only (control) or the GATA‐3 retrovector encoding both GATA‐3 and EGFP (GATA‐3).…”

Cell-Specific Transduction of Prdm1 -Expressing Lineages Mediated by a Receptor for Avian Leukosis Virus Subgroup B

“…To our knowledge, this is the first ALV-based system, and one of the few retroviral systems, allowing efficient retroviral transduction of in vivo-activated mature lymphocytes (21,24). Because only a minority of TVBexpressing cells are in cell cycle (e.g., most CD138 ϩ cells are quiescent plasma cells), the fraction of susceptible target cells transduced under the conditions delineated in this paper is likely to be significantly higher than the proportion (5%) of total cells transduced (Fig.…”