转化生长因子β1基因修饰的小鼠树突状细胞在移植体内的迁移？…

目的 观察β型转化生长因子（ＴＧＦβ１）基因修饰的Ｂ１０小鼠骨髓树突关细胞（ＤＣ）在Ｃ３Ｈ小鼠体内的迁移和存活,以探讨ＤＣ生物学特性与移植耐受性的关系。方法 从Ｂ１０小鼠骨髓分离ＤＣ前体细胞,在粒细胞－巨噬细胞集落刺激因子＋β型转化生长因子（ＧＭ－ＣＳＦ＋ＴＧＦβ）条件下培养增殖（ＧＣ１）并转导Ａｄ－ＬａｃＺ（Ｄｃ２）或Ａｄ－ＴＧＦβ１（ＤＣ３）,每组５×１０＾５个细胞注入Ｇ３Ｈ小鼠后掌皮下,分别

In vivo migration and survival of donor-derived dendritic cell progenitors genetically modified using an adenoviral vector encoding cDNA for TGF beta 1

OBJECTIVE: To investigate the migration and survival of B10 mouse bone marrow (BM)-derived DCs in C3H mice and if genetic modification of these DCs to overexpress TGF beta 1 may potentiate their tolerogenicity. METHOD: B10 mouse BM-derived DCs were propagated in GM-CSF and TGF beta(DC1), transduced DC1 with replication-deficient Ad vectors encoding genes for LacZ (DC2) or TGF beta 1(DC3). Cells of different groups were injected into one footpad of C3H mice. The mice were sacrificed on days 1, 2, 7, and 14, and spleens, thymuses, popliteal and mesenteric lymph nodes removed and stained with anti-IAb mAb. The incidences of B10 DC were determined by the mean number of IAb positive cells with dendriform morphology per low power field (x 100). RESULTS: Transduction with Ad-LacZ or Ad-TGF beta 1 did not affect DC migration or distribution in C3H recipients, i.e. IAb+ cells were first observed under the capsule of popliteal LN (peak at d1), then migrated into the marginal T dependent area of spleens (peak at d7), and were found occasionally in the thymus. Transduction of Ad-LacZ reduced the numbers of IAb+ cells identified in both LN and spleens at all time points post injection, compared with injection of unmodified control DC, suggesting that Ad transduction itself can affect DC life span in allogeneic recipients. Overexpression of TGF beta 1 by transduction of Ad-TGF beta 1 not only fully reversed the reduction of DC numbers induced by Ad transduction, but also prolonged the life span of DC in spleen, as shown by the 2-fold increase in number of IAb+ cells in spleen at d14 compared with control DCs. CONCLUSION: Mouse BM-derived TGF beta DCs can be transduced to express TGF beta 1 using an adenoviral vector, and exhibit the same migration characteristics as unmodified DC. The survival of TGF beta gene transduced DCs appears to be enhanced compared with unmodified or LacZ gene-transduced DCs.