PMID- 9322092
OWN - NLM
STAT- MEDLINE
DCOM- 19971105
LR  - 20171116
IS  - 1043-0342 (Print)
IS  - 1043-0342 (Linking)
VI  - 8
IP  - 13
DP  - 1997 Sep 1
TI  - Sustained high-level reconstitution of the hematopoietic system by preselected 
      hematopoietic cells expressing a transduced cell-surface antigen.
PG  - 1595-604
AB  - Despite improvements in retrovirus-mediated gene transfer to primitive murine 
      hematopoietic cells, high-level reconstitution with provirally marked cells with 
      continued expression of the transferred gene(s) remains a challenge in many 
      situations. We evaluated a physical preselection strategy for isolating 
      transduced cells after their infection with different vectors. The small (240-bp) 
      cDNA coding region for the human CD24 cell-surface antigen was inserted into 
      myeloproliferative sarcoma virus (MPSV) and murine stem cell virus (MSCV)-based 
      retroviral vectors such that expression of CD24 was under the control of the 
      viral long terminal repeat (LTR). After infection of (Ly-5.1) mouse bone marrow 
      (BM), those expressing CD24 were isolated by fluorescence-activated cell sorting 
      (FACS) and a transplant dose estimated to contain approximately 12 +/- 4 
      long-term competitive repopulating cells (CRU) injected into lethally irradiated 
      congenic Ly-5.2 recipients. Six months later, virtually all recipients showed 
      high-level (> 80%) reconstitution of their BM and thymus with Ly-5.1 
      (transplant-derived) cells, the majority of which were also transduced (mean of 
      2.5 or 2.6 proviral copies for the two vectors). All spleen colonies generated in 
      secondary recipients of cells obtained from the BM of the 6-month-old primary 
      mice contained the provirus. However, only in recipients of MSCVCD24-infected 
      marrow was a correspondingly high level of CD24 expression seen: a maximum of 88% 
      for whole BM (all mice positive), 58% for peripheral blood leukocytes (all mice 
      positive), and 21% for thymocytes (11 of 13 mice positive). CD24 was also readily 
      detected on the regenerated Sca-1+Lin- cells present in the primary and secondary 
      recipients when these were studied 6 months post-transplant, but again on more of 
      the Sca-1+Lin- cells in recipients of MSCVCD24-infected cells as compared to 
      recipients of MPSVCD24-infected cells. These results point to the utility of 
      preselection strategies and suggest an approach for the development of better 
      vectors for achieving regulated, lineage-specific or stage-specific gene 
      expression patterns in particular subsets of hematopoietic cells.
FAU - Pawliuk, R
AU  - Pawliuk R
AD  - Terry Fox Laboratory, British Columbia Cancer Agency, Vancouver, Canada.
FAU - Eaves, C J
AU  - Eaves CJ
FAU - Humphries, R K
AU  - Humphries RK
LA  - eng
GR  - P01 HI 55435-02/HI/NHLBI NIH HHS/United States
GR  - R01 HL48374-01/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Hum Gene Ther
JT  - Human gene therapy
JID - 9008950
RN  - 0 (Antigens, CD)
RN  - 0 (CD24 Antigen)
RN  - 0 (CD24 protein, human)
RN  - 0 (Cd24a protein, mouse)
RN  - 0 (Membrane Glycoproteins)
SB  - IM
MH  - Animals
MH  - Antigens, CD/*genetics
MH  - Bone Marrow Cells/*immunology
MH  - CD24 Antigen
MH  - Cell Lineage/genetics
MH  - Female
MH  - Genetic Vectors
MH  - Hematopoiesis/*genetics/immunology
MH  - Humans
MH  - Male
MH  - *Membrane Glycoproteins
MH  - Mice
MH  - Mice, Inbred C3H
MH  - Mice, Inbred C57BL
MH  - Retroviridae/genetics
MH  - *Transduction, Genetic
EDAT- 1997/10/10 00:00
MHDA- 1997/10/10 00:01
CRDT- 1997/10/10 00:00
PHST- 1997/10/10 00:00 [pubmed]
PHST- 1997/10/10 00:01 [medline]
PHST- 1997/10/10 00:00 [entrez]
AID - 10.1089/hum.1997.8.13-1595 [doi]
PST - ppublish
SO  - Hum Gene Ther. 1997 Sep 1;8(13):1595-604. doi: 10.1089/hum.1997.8.13-1595.