Molecular Characterization of the Element Controlling the Genetic Restriction of the B-B Cellular Interaction.

控制 B-B 细胞相互作用遗传限制的元素的分子表征。

• 批准号: 01570271
• 负责人: YAMAMOTO Hiroshi
• 金额: $ 1.34万
• 依托单位: National Institute of Neuroscience, National Center of Neurology and Psychiatry (1990)Kochi Medical School (1989)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01570271/
• 关键词: Idiotype; Idiotype-network; B lymphocytes; Long-term culture; Genetic restriction; Immunoglobulin gene; 遺伝子拘束性; マウスB細胞; B細胞クロン; B-B細胞間相互作用

Immunization of BALB/c mice with MOPC104E myeloma protein induces antiidiotypic B lymphocytes that have idiotype-specific enhancing activity on antibody production. The B-B cell interaction was restricted to both Igh and class II MHC. However, anti-Thy-1 and complement treated splenic B cells were maintained for more than 1 year in a mixture of concanavalin-A-stimulated splenocyte culture supernatand and synthetic medium. In applying the long term culture method, we have established a cloned B cell line named B19-1^d. B19-1^d cell are specific to MOPC104E or J558 cross-reactive idiotype and they express surface mu, lambda but no Ly-1. B19-1^d do not spontaneously secrete immunoglobulin but produce them upon stimulation with bacterial lipopolysaccharides. The effect of B19-1^d cell line on idiotypic antibody production was tested. Addition of only 10 to 100 B19ー1^d cells into dextran immune B cell culture greatly enhanced the idiotype-positive antidextran antibody responses. On the contrary, the antidextran antibody production was suppressed by the higher doses of B19-1^d cells. The effective cooperation between dextran-immune B cells and B19-1^d cloned B cells was restricted to class II MHC. By fusing lipopolysaccharide-stimulated B19-1^d clone with P3U1 myeloma, antiidiotypic antibody producing hybridoma was established.Each hybridoma cDNA and MOPC104E genomic DNA was cloned and their nucleic acid sequences were determined. The sequences were highly similar each other (91% similarity) and it was suggested that the both genes were derived from closely linked germ line genes.

用 MOPC104E 骨髓瘤蛋白免疫 BALB/c 小鼠可诱导抗独特型 B 淋巴细胞，其对抗体产生具有独特型特异性增强活性。B-B 细胞相互作用仅限于 Igh 和 II 类 MHC，然而，抗 Thy-1 和补体处理的脾脏。在长期应用中，B细胞在伴刀豆球蛋白-A刺激的脾细胞培养上清液和合成培养基的混合物中维持1年以上。通过培养方法，我们建立了一个名为 B19-1^d 的克隆 B 细胞系，该细胞对 MOPC104E 或 J558 交叉反应独特型具有特异性，并且表达表面 mu、lambda，但不表达 B19-1。 ^d不自发分泌免疫球蛋白，但在用细菌脂多糖刺激时产生免疫球蛋白。测试了B19-1^d细胞系对独特型抗体产生的影响。仅将10至100个B19-1^d细胞添加到葡聚糖免疫B细胞培养物中就大大增强了独特型阳性抗葡聚糖抗体反应，相反，较高剂量的B19-1^d细胞抑制了抗葡聚糖抗体的产生。葡聚糖免疫B细胞和B19-1^d克隆B细胞之间的有效合作仅限于II类MHC通过融合脂多糖刺激。建立了具有P3U1骨髓瘤的B19-1^d克隆，产生抗独特型抗体的杂交瘤。克隆了每个杂交瘤cDNA和MOPC104E基因组DNA，并确定了它们的核酸序列彼此高度相似（相似性为91%）。这两个基因都源自紧密相连的种系基因。

项目成果

Bitoh,S;Fujimoto,S;Yamamoto,H: "Idiotypic and anti-idiotypic B-B cell interaction is controlled by major histocompatibility complex-restricted regulation." Immunology. 66. 479-484 (1989)

Bitoh,S;Fujimoto,S;Yamamoto,H：“独特型和抗独特型 B-B 细胞相互作用是由主要组织相容性复合体限制性调节控制的。”

T.Tomoda: "Fluctuation of gene expression for polyー(ADPーribose)synthetase during heminーinduced erythroid differentiation of human leukemia K562 cells and its reversion process." Biochimica Biophysica Acta. (1991)

T. Tomoda：“血红素诱导的人类白血病 K562 细胞红系分化过程中聚（ADP-核糖）合成的基因表达波动及其逆转过程。”

N.Maeda: "Augmentation of human cytotoxic T lymphocytes against autologous tumor by a factor released from human monocytic leukemia cell line." Japanese Journal of Cancer Research. 80. 537-545 (1989)

N.Maeda：“通过人类单核细胞白血病细胞系释放的因子增强人类细胞毒性 T 淋巴细胞对抗自体肿瘤的能力。”

Taniguchi,T et al: "Requirement of down-regulation of NAD^+ ADP-ribosyl-transferase for the interferon-γ-induced activation precess of murine macrophage tumor cells." European Journal of Biochemistry.

Taniguchi, T 等人：“干扰素 γ 诱导的小鼠巨噬细胞肿瘤细胞激活过程需要下调 NAD^+ ADP-核糖基转移酶。”

S.Bitoh: "Evidence for idiotypicーand antiーidiotypic BーB cellular interaction with the use of cloned antiーidiotypic B cell line." Journal of Immunology. 144. 2046-2052 (1990)

S. Bitoh：“使用克隆的抗独特型 B 细胞系进行独特型和抗独特型 B-B 细胞相互作用的证据。”《免疫学杂志》144。2046-2052 (1990)