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production, with the aim of characterizing the phenotype of populations of cells
expressing the transgene in more detail. We found that 61% of eGFP-positive
cells were CD11b-positive,5% of these cells appeared to be CD14+ monocytes,
14% were CD20+ B cells and 10% were CD3+ T cells, 23% of which expressed
CD8 and 77% expressed CD4 (data not shown).
Figure 6. Flow cytometry analysis of hematopoiesis reconstitution. Animal transplanted with autologous CD34 +
bone marrow cells transduced with an SIV-based vector. eGFP-positive cells present in P1 and P2 were analyzed
by immuno-staining to identify the subpopulations of eGFP-positive cells in peripheral blood. CD20-PerCP-
Cy5, CD14-PE, CD11b-APC and CD3-APC staining were used to identify the B-lymphocyte, monocyte,
granulocyte and T-lymphocyte subpopulations.
discussion
The aim of this work was to study reconstitution of the myeloid and lymphoid
compartments after the autologous transplantation of genetically modified CD34 +
bone marrow cells into cynomolgus macaques previously subjected to gamma ir-
radiation.
We first assessed, in vitro, the efficiency with which a SIVmac251-derived
vector transduced macaque CD34+ hematopoietic bone marrow cells. These vec-
tors are similar to those derived from HIV. However, SIV-derived vectors clearly
outperform HIV-derived vectors in simian cells. In fact, HIV-1 fails to replicate
in simian cells because of an early postentry block [43,44], and Kootstra et., al
showed that the viral determinant involved in postentry restriction of HIV-1
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