Efficient transduction of murine B lymphocytes and B lymphoma lines by modified adenoviral vectors: enhancement via targeting to FcR and heparan-containing proteins

被引:13
作者
Li, L
Wickham, TJ
Keegan, AD
机构
[1] Amer Red Cross, Jerome H Holland Lab, Dept Immunol, Rockville, MD 20855 USA
[2] GenVec Inc, Gaithersburg, MD USA
关键词
B lymphocytes; adenovirus; transduction;
D O I
10.1038/sj.gt.3301487
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Murine lymphocytes are relatively refractory to efficient transfection or retroviral gene transduction. Adenovirus has been used as a vector to transduce a wide variety of cell types. Several advantages of adenoviruses are their ability to transduce non-cycling cells and to transduce the majority of cells in a population. Unfortunately, lymphocytes are not susceptible to infection with conventional adenovirus. Therefore, to express genes efficiently in murine B cells, we tested the ability of genetically modified adenovirus to transduce the p-galactosidase gene. We found that adenovirus containing polylysine in the fiber knob was able to efficiently transduce lipopolysaccharide (LPS)-activated splenic B cells and the B lymphoma line M12.4.1; greater than 80% of the cells expressed beta -galactosidase activity However, small resting S cells did not express activity unless treated with LPS after infection. This transduction was mediated by interaction with charged molecules since heparan-sulfate, and to a lesser degree chondroitan sulfate, inhibited the transduction. In addition, adenovirus containing a FLAG epitope in the fiber protein was used to target the FcR expressed on B cells using an anti-FLAG antibody. In the presence of anti-FLAG, the modified adenovirus was able to efficiently transduce LPS-activated B cells and several B cell lymphoma lines. interestingly in the absence of anti-FLAG, there was low level transduction in the LPS-blasts and in M12.4.1 that was not inhibited by soluble adenovirus fiber protein or agents that block RGD-integrin interactions. These results demonstrate that modified adenovirus efficiently transduce B lymphoctyes which will be critical for targeting genes to normal or malignant B cells.
引用
收藏
页码:938 / 945
页数:8
相关论文
共 34 条
[1]   CYTOPLASMIC DOMAIN HETEROGENEITY AND FUNCTIONS OF IGG FC-RECEPTORS IN LYMPHOCYTES-B [J].
AMIGORENA, S ;
BONNEROT, C ;
DRAKE, JR ;
CHOQUET, D ;
HUNZIKER, W ;
GUILLET, JG ;
WEBSTER, P ;
SAUTES, C ;
MELLMAN, I ;
FRIDMAN, WH .
SCIENCE, 1992, 256 (5065) :1808-1812
[2]   Isolation of a common receptor for coxsackie B viruses and adenoviruses 2 and 5 [J].
Bergelson, JM ;
Cunningham, JA ;
Droguett, G ;
KurtJones, EA ;
Krithivas, A ;
Hong, JS ;
Horwitz, MS ;
Crowell, RL ;
Finberg, RW .
SCIENCE, 1997, 275 (5304) :1320-1323
[3]   The murine CAR homolog is a receptor for coxsackie B viruses and adenoviruses [J].
Bergelson, JM ;
Krithivas, A ;
Celi, L ;
Droguett, G ;
Horwitz, MS ;
Wickham, T ;
Crowell, RL ;
Finberg, RW .
JOURNAL OF VIROLOGY, 1998, 72 (01) :415-419
[4]   Polylysine modification of adenoviral fiber protein enhances muscle cell transduction [J].
Bouri, K ;
Feero, WG ;
Myerburg, MM ;
Wickham, TJ ;
Kovesdi, I ;
Hoffman, EP ;
Clemens, PR .
HUMAN GENE THERAPY, 1999, 10 (10) :1633-1640
[5]   Adenovirus vector infection of chronic lymphocytic leukemia B cells [J].
Cantwell, MJ ;
Sharma, S ;
Friedmann, T ;
Kipps, TJ .
BLOOD, 1996, 88 (12) :4676-4683
[6]   The Mi15 monoclonal antibody (anti-Syndecan-1) is a reliable marker for quantifying plasma cells in paraffin-embedded bone marrow biopsy specimens [J].
Costes, V ;
Magen, V ;
Legouffe, E ;
Durand, L ;
Baldet, P ;
Rossi, JF ;
Klein, B ;
Brochier, J .
HUMAN PATHOLOGY, 1999, 30 (12) :1405-1411
[7]   The human HLA-A*0201 allele, expressed in hamster cells, is not a high-affinity receptor for adenovirus type 5 fiber [J].
Davison, E ;
Kirby, I ;
Elliott, T ;
Santis, G .
JOURNAL OF VIROLOGY, 1999, 73 (05) :4513-4517
[8]   Targeted gene delivery by tropism-modified adenoviral vectors [J].
Douglas, JT ;
Rogers, BE ;
Rosenfeld, ME ;
Michael, SI ;
Feng, MZ ;
Curiel, DT .
NATURE BIOTECHNOLOGY, 1996, 14 (11) :1574-1578
[9]   High-efficiency gene transfer into ex vivo expanded human hematopoietic progenitors and precursor cells by adenovirus vectors [J].
Frey, BM ;
Hackett, NR ;
Bergelson, JM ;
Finberg, R ;
Crystal, RG ;
Moore, MAS ;
Rafii, S .
BLOOD, 1998, 91 (08) :2781-2792
[10]   Expression of alpha v and beta 3 integrin chains on murine lymphocytes [J].
Gerber, DJ ;
Pereira, P ;
Huang, SY ;
Pelletier, C ;
Tonegawa, S .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (25) :14698-14703