Immunochemical and genetical studies on B cell abnormality in patients with immunoglobulin deficiencies

从免疫球蛋白缺乏症患者中提取B细胞，进行免疫化学和遗传学研究：（1）免疫球蛋白产生功能分析;（2）膜糖蛋白天冬酰胺连接糖链分析;（3）免疫球蛋白产生基因分析。所得结果如下。(1)免疫球蛋白产生功能;使用商陆有丝分裂原、爱泼斯坦巴尔病毒和科万病毒研究B细胞或T细胞功能。作为结果，很明显，在许多患者中B细胞功能受到干扰，而且患者的B细胞功能障碍存在很大的异质性。(2)天冬酰胺连接的膜糖蛋白的糖链;这是通过肼解和连续外切糖苷酶消化的方法研究的。结果表明，在B细胞中，分化的B细胞具有更多的具有二分型N-乙酰氨基葡萄糖（GlcNAc）的双触角糖链，更多的具有GlcNAc 2->4 Man结合的三触角糖链和更多的四触角糖链。T细胞不具有具有二等分GlcNAc的双触角糖链。另一方面，健康B细胞和免疫缺陷B细胞在结构上没有显著差异。这些结果表明，具有二等分GlcNAc的双触角糖链与B细胞功能有关。(3)免疫球蛋白产生基因：在健康淋巴细胞和免疫缺陷淋巴细胞上，用硫氰酸胍法获得其mRNA，用细胞自由翻译法从mRNA合成免疫球蛋白。免疫印迹等方法检测免疫球蛋白，结果表明，正常人和免疫缺陷者均合成IgM。从这个结果可以看出，在某些患者中，mRNA水平后的途径可能在免疫球蛋白合成中受到干扰。

Abnormalities of B cells from patients with immunoglobulin deficiencies wewe immunochemically and genetically investigated in points of (1) analysis of immunoglobulin productive function (2) analysis of asparagine linked sugar chains of membrane glycoproteins (3) analysis of gene of immunoglobulin production. The results obtained were as follow.(1) Immunoglobulin productive function; B cell or T cell function was investigated using Poke weed mitogen,Epstein Barr virus and Cowan l. As the results,it was clear that in many patients B cell function was disturbed,and moreover there was much heterogeneity in B cell dysfunctions of patients.(2) Asparagine linked sugar chain of membrane glycoprotein; This was studied by methods of hydrazinolysis and sequential exoglycosidase digestion. As the results,it was clear that,in B cells ,more differentiated B cells had the biantennary sugar chain with bisect N-acetylglucosamine(GlcNAc), more triantennary sugar chain with GlcNAc 2->4 Man bound and more tetraantennary sugar chains. T cells did not have the biantennary sugar chain with bisect GlcNAc. On the other hand,there was no significant difference between healthy B cells and immunodeficient B cells in structures. From these results, it is suggested that the biantennary sugar chain with bisect GlcNAc is concerning to B cell functions.(3) Gene of immunoglobulin production; On healthy lymphocytes and immunodeficient lymphocytes, their mRNA were obtained by the method of guanidium-thiocyanate,and immunoglobulin was synthesized by the method of cell free translation from the mRNA. The immunoglobulin was detected by the Western-blotting method and so on. As the results, IgM was synthesized in both healthy and immunodeficient persons. From this result,it is suggested that the pathway after mRNA level may be disturbed in immunoglobulin synthesis on certain patients.