期刊
MICROBIOLOGY AND IMMUNOLOGY
卷 54, 期 2, 页码 112-121出版社
WILEY
DOI: 10.1111/j.1348-0421.2009.00190.x
关键词
anti-prion effect; monoclonal antibody; single-chain fragment variable region
资金
- Ministry of Agriculture, Forestry and Fisheries of Japan
mAbs T1 and T2 were established by immunizing PrP gene ablated mice with recombinant MoPrP of residues 121-231. Both mAbs were cross-reactive with PrP from hamster, sheep, cattle and deer. A linear epitope of mAb T1 was identified at residues 137-143 of MoPrP and buried in PrPC expressed on the cell surface. mAb T1 showed no inhibitory effect on accumulation of PrPSc in cultured scrapie-infected neuroblastoma (ScN2a) cells. In contrast, mAb T2 recognized a discontinuous epitope ranged on, or structured by, residues 132-217 and this epitope was exposed on the cell surface PrPC. mAb T2 showed an excellent inhibitory effect on PrPSc accumulation in vitro at a 50% inhibitory concentration of 0.02 mu g/ml (0.14 nM). The scFv form of mAb T2 (scFv T2) was secreted in neuroblastoma (N2a58) cell cultures by transfection through eukaryotic secretion vector. Coculturing of ScN2a cells with scFv T2-producing N2a58 cells induced a clear inhibitory effect on PrPSc accumulation, suggesting that scFv T2 could potentially be an immunotherapeutic tool for prion diseases by inhibition of PrPSc accumulation.
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