Gene transfer using recombinant simian virus 40 viral vectors into mice bone marrow progenitor cells depressed the immunogenicity of keratinocyte stem cells.

Abstract:

:Hematopoietic stem cell (HSC) gene transfer has been attempted almost entirely ex vivo and has been limited by loss of self-renewal capacity and transplantation-related defects in homing and engraftment. Herein we have attempted to overcome these limitations by injecting vectors directly into the bone marrow (BM) to transduce HSCs in their native environment. Simian virus 40 (SV40)-derived gene delivery vectors were used because they efficiently transduce resting CD34+ cells. Neonatal C57BL/6 (H-2b) mice (3 days old) received SV(Nef-FLAG), carrying FLAG marker epitope directly into both femoral marrow cavities. Keratinocyte stem cells (KSCs) were purified at 7 and 14 days after SV40 injection. The KSCs from 10-day-old C57BL/6 mice were designated as controls. Flow cytometric (FCM) analyses indicated that KSCs from transgenic mice showed strong down-regulation of surface immunological molecules CD40, CD80, CD86, and human major histocompatibility complex class I chain-related antigen A (MICA). Mixed lymphocyte reaction (MLR) assays showed that transgenic KSCs depressed allogeneic T-cell proliferation. Immunofluorescence showed transgenic KSCs expressed FLAG for the entire study as well as high levels of transforming growth factor (TGF)-beta and BCL-2. Thus, direct intramarrow administration of recombinant SV40 yielded efficient gene transfer to mice BM progenitor cells. KSCs with low immunogenicity may be obtained for further investigations of skin transplantation immunity.

journal_name

Transplant Proc

authors

Li W,Xu T,Wang JF,Wu XF,Li M,Lu PY

doi

10.1016/j.transproceed.2008.06.090

subject

Has Abstract

pub_date

2008-12-01 00:00:00

pages

3656-9

issue

10

eissn

0041-1345

issn

1873-2623

pii

S0041-1345(08)01040-3

journal_volume

40

pub_type

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