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MR Vol.11 No.2 indexに戻る
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MODERN RHEUMATOLOGY
Vol.11 No.2 |
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"Inverse wrap": an improved
implantation technique for virus-transduced synovial fibroblasts
in the SCID mouse
model for rheumatoid arthritis |
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| M. Judex1, E. Neumann1, M. Fleck1, T.
Pap2, J. D. Mountz3, R. E. Gay2, J. Scholmerich1, K. Nishioka4,
S. Gay2, U. Muller-Ladner1 |
(1)Department of Internal Medicine I, University
of Regensburg, FJS-Allee 11, D-93042 Regensburg, Germany
(2)Department of Rheumatology, Center for Experimental Rheumatology,
University Hospital, Zurich, Switzerland
(3)Department of Rheumatology, University of Alabama at Birmingham, Birmingham,
AL, USA
(4)Institute of Medical Science, St. Marianna University, Kawasaki, Japan |
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| Abstract |
| Abstract The SCID mouse model for rheumatoid
arthritis (RA) is an established and reliable approach to examining
the distinct mechanisms operative in RA synovium, and evaluating
novel gene therapy strategies. However, serum concentrations of
circulating gene therapy products following gene transfer are frequently
too low to allow detection. This problem stimulated us to develop
a novel implantation technique to improve the yield of these soluble
gene products. Synovial fibroblasts from patients with RA were
cultured, passaged, and transduced with Ad5 sTNFRp55:Ig. sTNFRp55:Ig
production was confirmed by ELISA, and then cells were implanted
into SCID mice using a novel implantation strategy in which pieces
of human cartilage were engrafted into a fibroblast-saturated inert
sponge. Thereafter, the sponges were implanted under the skin of
the mice instead of under the kidney capsule, as in the original
approach, allowing co-implantation of larger pieces of cartilage
together with higher numbers of adenovirus-transduced RA synovial
fibroblasts. The improved implantation technique not only resulted
in a reduction in the number of mice needed in each experiment
by approximately 60%, and a reduction of the time taken for surgery
by about 50%, but also considerably enhanced the serum concentrations
of the gene product sTNFRp55-Ig, allowing detection of the soluble
TNF receptor p55 by standard ELISA. In summary, the improved implantation
technique for the SCID mouse model for RA results in more economic
animal treatment, and facilitates the detection and quantification
of circulating gene products following adenovirus-based gene transfer
into synovial fibroblasts. |
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| Key words |
| Key words Gene transfer ・ Inverse wrap ・ Rheumatoid
arthritis ・ SCID mouse ・ TNF receptor |
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