Kiefer R, Haas C A, Kreutzberg G W
Department of Neuromorphology, Max-Planck-Institute for Psychiatry, Martinsried, F.R.G.
Neuroscience. 1991;45(3):551-60. doi: 10.1016/0306-4522(91)90270-x.
Gamma interferon is a potent immunoregulatory peptide produced by activated lymphocytes. Recently, a gamma interferon-like immunoreactive molecule has been demonstrated immunohistochemically in subpopulations of rat neurons. We have now further characterized this molecule. Western blot analysis of spinal ganglia homogenates revealed a single 60,000 mol. wt band that was immunoreactive with monoclonal antibody DB1 directed against rat gamma interferon. A polyclonal antiserum and the monoclonal antibodies DB10 and DB12 failed to detect this band although all antibodies were able to label the major 18,000 mol. wt band of recombinant gamma interferon on the same blots. The 60,000 mol. wt band was selectively present in homogenates from primary sensory and sympathetic ganglia but was absent from the central nervous system and other peripheral organs, corresponding to the reported immunocytochemical distribution of gamma interferon-like immunoreactivity. The 60,000 mol. wt protein does not appear to be glycosylated. It could not be solubilized by detergents such as Triton X-100 and it co-purified with cytoskeleton-enriched preparations. At the nucleic acid level, Northern blot analysis using probes specific for rat gamma interferon mRNA failed to detect specific mRNA in rat spinal ganglia, whereas a strong 1.2 kb signal was detected in activated spleen cells. Functionally, gamma interferon-like immunoreactive material is strongly induced in superior cervical ganglion neurons after preganglionic axotomy of the sympathetic chain, but remains constant or slightly decreases in L5 spinal ganglion neurons after sciatic nerve transection. In contrast, major histocompatibility complex antigens are strongly induced on non-neuronal cells in both systems. We conclude that the neuronal gamma interferon-like immunoreactive material is clearly distinct from lymphocyte-derived gamma interferon and might not be involved in the control of major histocompatibility complex expression on glial cells.
γ干扰素是一种由活化淋巴细胞产生的强效免疫调节肽。最近，通过免疫组织化学方法在大鼠神经元亚群中证实了一种γ干扰素样免疫反应性分子。我们现在对该分子进行了进一步的特性分析。脊髓神经节匀浆的蛋白质免疫印迹分析显示有一条单一的60,000道尔顿分子量的条带，它与针对大鼠γ干扰素的单克隆抗体DB1发生免疫反应。尽管所有抗体都能够在同一张印迹上标记重组γ干扰素的主要18,000道尔顿分子量的条带，但一种多克隆抗血清以及单克隆抗体DB10和DB12未能检测到这条带。60,000道尔顿分子量的条带选择性地存在于初级感觉神经节和交感神经节的匀浆中，但在中枢神经系统和其他外周器官中不存在，这与报道的γ干扰素样免疫反应性的免疫细胞化学分布一致。60,000道尔顿分子量的蛋白质似乎未被糖基化。它不能被诸如 Triton X - 100 之类的去污剂溶解，并且与富含细胞骨架的制剂共同纯化。在核酸水平上，使用针对大鼠γ干扰素mRNA的特异性探针进行的Northern印迹分析未能在大鼠脊髓神经节中检测到特异性mRNA，而在活化的脾细胞中检测到一个强的1.2 kb信号。在功能上，交感神经链节前轴突切断后，颈上神经节神经元中γ干扰素样免疫反应性物质被强烈诱导，但坐骨神经切断后，L5脊髓神经节神经元中的该物质保持恒定或略有下降。相比之下，两个系统中的非神经元细胞上主要组织相容性复合体抗原均被强烈诱导。我们得出结论，神经元γ干扰素样免疫反应性物质明显不同于淋巴细胞来源的γ干扰素，并且可能不参与胶质细胞上主要组织相容性复合体表达的调控。
