PROTEIN KINASE ISOFORM ISOFORMS IN LYMPHOID CELLS.

淋巴细胞中的蛋白激酶同种型。

• 批准号: 06670476
• 负责人: TSUTSUMI Akito
• 金额: $ 1.34万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06670476/
• 关键词: Protein Kinase C; Isoform; Lymphocytes; PKCdelta; Somatostatin receptors

1) A Western blot system for detecting PKC isoforms in lymphoid cells was developed.2) Polymerase chain reaction (PCR) primers for PKC lambda rheta and eta were made. Reverse transcription-PCR (RT-PCR) with a mouse preB cell line 70z/3 and a mouse B cell line WEHI231 showed that PKC lambda and rheta mRNA is detectable in these cell lines.3) An expression vector for overexpression of PKCdelta was constructed using PKCdelta cDNA.The vector was transfected to 70z/3 cells and Jurkat human lymphoid cells. Unexpectedly, we could not detect overexpression of PKCdelta protein by Western blot analysis. The reason for this is now under investigation.4) Using the same methodology, somatostatin receptor (SSTR) subtype expression in human lymphoid cells was investigated. It was found that although there is heterogeneity in SSTR subtypes in lymphoid cells, SSTR2 is the most dominant subtype. It was also found that although human peripheral lymphoid cells express little or no SSTR2 mRNA,peripheral lymphocytes from patients with lymphatic leukemia may express abundant amount of SSTR2 mRNA.These results suggest that expression of SSTR2 may have a role in development, activation and tumorgenesis of human lymphoid cells.

1)建立了一种检测淋巴样细胞PKC亚型的Western blot系统。2)制作PKC lambda、rheta和eta的聚合酶链反应（PCR）引物。小鼠preB细胞系70z/3和小鼠B细胞系WEHI231的逆转录pcr结果显示，在这些细胞系中均可检测到PKC lambda和rheta mRNA。3)利用PKCdelta cDNA构建PKCdelta过表达载体。载体转染70z/3细胞和Jurkat人淋巴样细胞。出乎意料的是，我们无法通过Western blot检测到PKCdelta蛋白的过表达。原因正在调查中。4)采用相同的方法，研究了生长抑素受体（SSTR）亚型在人淋巴样细胞中的表达。结果发现，虽然淋巴样细胞中SSTR亚型存在异质性，但SSTR2是最占优势的亚型。我们还发现，虽然人类外周淋巴样细胞很少或不表达SSTR2 mRNA，但淋巴白血病患者的外周淋巴细胞可能表达大量的SSTR2 mRNA。这些结果提示SSTR2的表达可能在人淋巴样细胞的发育、激活和肿瘤发生中起作用。

项目成果

Tsutsumi A: "Pathogenic anticardiolipin antibodies in patients with SLE recognize epitopes expressed on beta2-Glycoprotein I." Lupus. 4. 42- (1995)

Tsutsumi A：“SLE 患者中的致病性抗心磷脂抗体可识别 β2-糖蛋白 I 上表达的表位。”

Amasaki Y: "Up-regulated expression of Fas antigen (CD95) by peripheral naive and memory T cell subsets in patients with systemic lupus erythematosus (SLE): a possible mechanism for lymphopenia." Clin Exp Immunol. 99. 245-250 (1995)

Amasaki Y：“系统性红斑狼疮 (SLE) 患者外周幼稚 T 细胞亚群和记忆 T 细胞亚群上调 Fas 抗原 (CD95) 表达：淋巴细胞减少症的可能机制。”

Koike T: "Antigenic specificity of the“anticardiolipin"antibodies." Blood. 85. 2277-2278 (1995)

Koike T：““抗心磷脂”抗体的抗原特异性。”《血液》。85. 2277-2278 (1995)

Koike T: "Pulmonary hypertension and the antiphospholipid syndrome." Internal Medicine. 34. 938 (1995)

Koike T：“肺动脉高压和抗磷脂综合征。”

Koike T,Tsutsumi A et al.: "Antigenic specificity of the "anticardiolipin" antibodies." Blood. 85. 2277-2278 (1995)

Koike T、Tsutsumi A 等人：“‘抗心磷脂’抗体的抗原特异性。”