Grawunder U, Haasner D, Melchers F, Rolink A
Basel Institute for Immunology, Switzerland.
Int Immunol. 1993 Dec;5(12):1609-18. doi: 10.1093/intimm/5.12.1609.
The kinetics of kappa light (kappa L) chain gene rearrangement and expression on mRNA and protein level has been studied with four stromal cell/IL-7 reactive, long-term in vitro proliferating pre-B cell lines and clones, two from fetal liver of normal mice and two from fetal liver of E microH-bcl-2 transgenic (bcl-2-tg) mice. These pre-B cell lines and clones are DJH-rearranged on both H chain alleles. Two of the clones harbor H chain rearrangements which do not allow the expression of VHDJH rearranged H chain genes as microH chain proteins. Upon removal of IL-7 from the pre-B cell cultures all four cell lines rearrange VH-DJH and VL-JL gene segments, loose the surface expression of c-kit, CD43, and surrogate light chain, as well as the capacity to be clonable on stromal cells in the presence of IL-7. Pre-B cells from normal mice die by apoptosis during differentiation, while those from bcl-2-tg mice do not. All four lines and clones express comparable levels of mRNA for microH and kappa L chains with the same time kinetics during 3 days of differentiation. However, only two of the four pre-B cell lines and clones express microH chain protein, whereas all four pre-B cell lines and clones express kappa L chain protein at comparable levels between 2 x 10(5) and 1.4 x 10(6) kappa L chain molecules per cell. These results suggest that microH chain expression is not mandatory for rearrangement and normal expression of kappa L chain genes when pre-B cells differentiate to B cells.
利用四个基质细胞/IL-7反应性、长期体外增殖的前B细胞系和克隆,研究了κ轻链(κL)基因重排以及在mRNA和蛋白质水平上的表达情况,其中两个来自正常小鼠的胎肝,另外两个来自EμH-bcl-2转基因(bcl-2-tg)小鼠的胎肝。这些前B细胞系和克隆在两条重链等位基因上均发生了DJH重排。其中两个克隆具有重链重排,不允许VHDJH重排的重链基因作为μ重链蛋白表达。从前B细胞培养物中去除IL-7后,所有四个细胞系均重排VH-DJH和VL-JL基因片段,失去c-kit、CD43和替代轻链的表面表达,以及在IL-7存在下在基质细胞上克隆的能力。正常小鼠的前B细胞在分化过程中通过凋亡死亡,而bcl-2-tg小鼠的前B细胞则不会。在分化的3天内,所有四个细胞系和克隆表达的μ重链和κL链mRNA水平相当,且具有相同的时间动力学。然而,四个前B细胞系和克隆中只有两个表达μ重链蛋白,而所有四个前B细胞系和克隆均以每细胞2×10⁵至1.4×10⁶个κL链分子的相当水平表达κL链蛋白。这些结果表明,当前B细胞分化为B细胞时,μ重链表达对于κL链基因的重排和正常表达并非必需。