Yamamura K, Kudo A, Ebihara T, Kamino K, Araki K, Kumahara Y, Watanabe T
Proc Natl Acad Sci U S A. 1986 Apr;83(7):2152-6. doi: 10.1073/pnas.83.7.2152.
A functionally rearranged human gamma 1 heavy-chain immunoglobulin gene was cloned from a human plasma cell leukemia cell line, ARH-77, into the phage lambda Charon 4A. The recombinant phage DNA was introduced into fertilized mouse eggs (about 200 copies of the human gene per egg). A total of 30 mice were born and were screened for the presence of the human gamma 1 gene by dot hybridization. Two of these 30 mice had integrated one or two copies of the gene. The gamma 1 mRNAs were detected only in spleen. Levels of gamma 1 mRNA and the percentage of spleen cells producing human gamma chain increased up to 50-fold after treatment with bacterial lipopolysaccharide (a B-cell mitogen) but not with concanavalin A (a T-cell mitogen), suggesting B-cell-specific and regulated expression of the human gamma 1 heavy-chain gene. Human gamma chain-producing cells were found only in the periphery of the germinal center of the white pulp in histological sections of the spleen but not in sections of other tissues. Human gamma chains appeared to be coupled with mouse light chains to form a complete IgG molecule and were secreted into the cell supernatant. The production and secretion of endogenous immunoglobulin heavy and light chains in transgenic mice appeared to be the same as in normal mice. About one-seventh of the spleen cells that produced endogenous mouse heavy chains also produced human gamma chains, but no cells that produced only human gamma chain were observed.
从人浆细胞白血病细胞系ARH - 77中克隆出功能重排的人γ1重链免疫球蛋白基因，并将其插入噬菌体λCharon 4A中。将重组噬菌体DNA导入受精的小鼠卵（每个卵约含200份人基因拷贝）。共出生30只小鼠，通过点杂交筛选人γ1基因的存在情况。这30只小鼠中有2只整合了一或两份该基因。γ1 mRNA仅在脾脏中检测到。用细菌脂多糖（一种B细胞有丝分裂原）处理后，γ1 mRNA水平和产生人γ链的脾细胞百分比增加了50倍，而用伴刀豆球蛋白A（一种T细胞有丝分裂原）处理则无此现象，这表明人γ1重链基因具有B细胞特异性且受调控表达。在脾脏组织切片中，仅在白髓生发中心的周边发现产生人γ链的细胞，而在其他组织切片中未发现。人γ链似乎与小鼠轻链结合形成完整的IgG分子，并分泌到细胞上清液中。转基因小鼠内源性免疫球蛋白重链和轻链的产生和分泌似乎与正常小鼠相同。约七分之一产生内源性小鼠重链的脾细胞也产生人γ链，但未观察到仅产生人γ链的细胞。
