Characterization of cells with T cell markers in athymic nude bone marrow and of their in vitro-derived clonal progeny. Comparison with euthymic bone marrow.
Characterization of cells with T cell markers in athymic nude bone marrow and of their in vitro-derived clonal progeny. Comparison with euthymic bone marrow.
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无胸腺裸骨髓中具有 T 细胞标记的细胞及其体外克隆后代的特征。
DOI：
10.4049/jimmunol.144.2.411
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发表时间：
1990
影响因子：
4.4
通讯作者：
Jörg Reimann
中科院分区：
文献类型：
--
作者：
P. Benveniste;B. Chadwick;Richard G. Miller;Jörg Reimann
有证据表明，T细胞分化的早期步骤，TCR表达的点，可以发生在无胸腺裸骨髓（BM），也许也在正常的BM。5 ~ 8周龄裸鼠和正常小鼠的低密度尼龙毛非贴壁BM中含有暗淡的CD 3 + CD 4-CD 8-细胞。这个亚群在scid/scid低密度尼龙羊毛非粘附BM中没有发现，但在裸BM和正常BM中以相似的频率存在，并且表达了相当的荧光强度。通过使用额外的标记物，发现裸和正常dim CD 3 + CD 4-CD 8-细胞之间的差异：裸中J11 d +/J11 d-细胞的比率比正常BM中高至少3倍。尽管60%的裸J11 d + CD 3+细胞表达TCR-α/β，40%表达γ/δ TCR，但在J11 d+或J11 d-亚群的正常小鼠的dim CD 3+细胞中几乎没有检测到TCR-γ/δ表达细胞。这些细胞中的大多数表达TCR-α/β。我们建立了裸暗CD 3 + CD 4-CD 8-细胞与成熟再循环T细胞的区别，并开发了一种支持这些细胞生长的体外克隆培养系统。当这些细胞在有限稀释系统中与灭活的抗CD 3 mAb分泌细胞体外共培养时，在rIL-2和rIL-4存在下，5 × 10（2）至1 × 10（3）个细胞中有1个细胞是生长诱导型的。大多数子代细胞表达CD 3 + CD 8 + CD 4-表面表型。培养物显示非特异性细胞溶解反应性。使用敏感的斑点印迹技术，在这些培养物中检测到编码C α、C β、VC γ和C δ TCR基因的转录本。再杂交实验表明，在大多数克隆中，C α或C δ mRNA转录物的相互排斥的表达被选择用于克隆性的高概率。大多数（大于95%）来源于正常BM的T细胞克隆表达C α转录物，其中约5%表达C δ转录物。相比之下，约40%的T细胞克隆来源于裸BM表达C δ转录本，而剩余的60%表达比正常BM来源的克隆低10倍的C α信使。
Evidence is presented that early steps in T cell differentiation, to the point of TCR expression, can occur in athymic nude bone marrow (BM) and perhaps also in normal BM. Low density nylon wool nonadherent BM from 5- to 8-wk-old nude and normal mice was found to contain dim CD3+ CD4- CD8- cells. This subpopulation was not found in scid/scid low density nylon wool nonadherent BM but was present at similar frequency in both nude and normal BM and expressed comparable fluorescence intensity. With the use of additional markers, a difference between nude and normal dim CD3+ CD4- CD8- cells was found: the ratio of J11d+/J11d- cells was at least threefold higher in nude than in normal BM. Whereas 60% of nude J11d+ CD3+ cells expressed a TCR-alpha/beta and 40% expressed a gamma/delta TCR, almost no TCR-gamma/delta expressing cells were detected in dim CD3+ cells from normal mice in either the J11d+ or J11d- subpopulation. The majority of those cells express a TCR-alpha/beta. We established that nude dim CD3+ CD4- CD8- cells were distinguishable from mature recirculating T cells and developed an in vitro clonal culture system that supported the growth of these cells. When these cells were cocultured in vitro in a limiting dilution system with inactivated anti-CD3 mAb-secreting cells, in the presence of rIL-2 and rIL-4, one in 5 x 10(2) to 1 x 10(3) cells were growth inducible. Most progeny cells expressed a CD3+ CD8+ CD4- surface phenotype. Cultures displayed nonspecific cytolytic reactivity. With the use of a sensitive dot-blot technique, transcripts coding for C alpha, C beta, VC gamma, and C delta TCR genes were detected in these cultures. Rehybridization experiments demonstrated mutually exclusive expression of either C alpha or C delta mRNA transcripts in the majority of clones selected for a high probability of clonality. Most (greater than 95%) of T cell clones derived from normal BM expressed the C alpha transcript with approximately 5% expressing the C delta transcript. In contrast, about 40% of T cell clones derived from nude BM expressed the C delta transcript whereas the remaining 60% expressed 10-fold lower C alpha message than normal BM-derived clones.