Recent studies have revealed that the γ-chain of theIL-2 receptor is shared by the receptors for IL-4, IL7, IL-9, IL-13, and IL-15, and it is therefore also referred toas the common γ-chain (γc). Mutations of γc r...Recent studies have revealed that the γ-chain of theIL-2 receptor is shared by the receptors for IL-4, IL7, IL-9, IL-13, and IL-15, and it is therefore also referred toas the common γ-chain (γc). Mutations of γc result inX-linked severe combined immunodeficiency syndrome inhumans, indicating that rye is essential for normal development and function of the immune system. We demonstratethat human hematopoietic cells express two γc transcriptsdiffering in their carboxyl terminal coding region. Onetranscript is the previously reported sequence (γc-long),whereas the newly identified sequence exhibits a deletion of72 nucleotides close to the 3’-end of the open reading frame(γc-short). This alteration predicts a loss of 24 amino acidsincluding a conserved tyrosine residue which is shared byseveral members of the cytokine receptor family. Thepresence of these two distinct forms of rye transcripts wasdemonstrated by sequencing of reversely transcribed andpolymerase chain reaction (RT-PCR) amplified mRNA, restriction digestion of the RT-PCR products, RNAse protection, and Northern blotting from human cell lines andhuman peripheral blood lymphocytes. Furthermore, thetwo variants were present in peripheral blood lymphocytesfrom both female and male donors, which rules out allelicvariants since rye is a single copy gene located on the Xchromosome. A truncation mutant at a site near the observed changes in γc-short has been reported by othersto alter biochemical events activated by cytokines. Thiscombined with the loss of a potential SH2 "docking" sitein γc-short suggests that γc-long and γc-short may link todifferent signaling pathways and may play an importantrole in determining the cellular response to IL-2, IL-4, IL-7, IL-9, IL-13, IL-15.展开更多
为了建立稳定表达猪FcγRⅢ(Porcine Fc gamma receptorⅢ,poFcγRⅢ)的Marc-145细胞系,从PAM细胞中提取总RNA,应用RT-PCR技术获得猪FcγRⅢ和γ链的cDNA,并构建PIREShyg3-γ和pcDNA3.1-FcγRⅢ真核表达质粒;用脂质体共转染Marc-145细胞...为了建立稳定表达猪FcγRⅢ(Porcine Fc gamma receptorⅢ,poFcγRⅢ)的Marc-145细胞系,从PAM细胞中提取总RNA,应用RT-PCR技术获得猪FcγRⅢ和γ链的cDNA,并构建PIREShyg3-γ和pcDNA3.1-FcγRⅢ真核表达质粒;用脂质体共转染Marc-145细胞,经潮霉素B(300 mg/L)和G418(400 mg/L)共筛选获得稳定表达猪FcγRⅢ的细胞系;运用RT-PCR、玫瑰花环试验和流式细胞术对细胞系进行了鉴定。结果表明,成功构建了猪FcγRⅢ和γ链真核表达载体,建立了稳定表达猪FcγRⅢ的细胞系,表达于转染细胞表面的猪FcγRⅢ受体分子能与猪IgG特异结合。展开更多
Objective:To study the role of IL 2Rγ chain in the immuno senescence process Methods:8 young, aged from 18 to 35 years(average 24 9±5 8 years), and 17 old healthy people, aged from 65 to 79 years(average 69 4...Objective:To study the role of IL 2Rγ chain in the immuno senescence process Methods:8 young, aged from 18 to 35 years(average 24 9±5 8 years), and 17 old healthy people, aged from 65 to 79 years(average 69 4±4 2 years), had been selected for determination the proliferative responses of peripheral blood lymphocytes to PHA, the percentage and dynamic changes of CD132, CD45R positive cells, also, the gene expression of IL 2Rγ chain with RT PCR using the designed primers Results:The results showed that the PHA proliferative response of lymphocytes of aged people(0 199±0 055) were lower than that of young people(0 279±0 054), P=0 0012 The expression of IL 2Rγ chain on the lymphocytes surface decreased in aged people, 46 2%±7 7%, compared to young people, 68 0%±11 8%, P<0 001, It was verified from RT PCR result that the event occurred at transcription level The time dynamic curve of IL 2Rγ chain expression were different between aged and young people Conclusion:A preliminary conclusion would be drawn that the IL 2Rγ chain play an important role in the declined immune function of aged展开更多
文摘Recent studies have revealed that the γ-chain of theIL-2 receptor is shared by the receptors for IL-4, IL7, IL-9, IL-13, and IL-15, and it is therefore also referred toas the common γ-chain (γc). Mutations of γc result inX-linked severe combined immunodeficiency syndrome inhumans, indicating that rye is essential for normal development and function of the immune system. We demonstratethat human hematopoietic cells express two γc transcriptsdiffering in their carboxyl terminal coding region. Onetranscript is the previously reported sequence (γc-long),whereas the newly identified sequence exhibits a deletion of72 nucleotides close to the 3’-end of the open reading frame(γc-short). This alteration predicts a loss of 24 amino acidsincluding a conserved tyrosine residue which is shared byseveral members of the cytokine receptor family. Thepresence of these two distinct forms of rye transcripts wasdemonstrated by sequencing of reversely transcribed andpolymerase chain reaction (RT-PCR) amplified mRNA, restriction digestion of the RT-PCR products, RNAse protection, and Northern blotting from human cell lines andhuman peripheral blood lymphocytes. Furthermore, thetwo variants were present in peripheral blood lymphocytesfrom both female and male donors, which rules out allelicvariants since rye is a single copy gene located on the Xchromosome. A truncation mutant at a site near the observed changes in γc-short has been reported by othersto alter biochemical events activated by cytokines. Thiscombined with the loss of a potential SH2 "docking" sitein γc-short suggests that γc-long and γc-short may link todifferent signaling pathways and may play an importantrole in determining the cellular response to IL-2, IL-4, IL-7, IL-9, IL-13, IL-15.
文摘Objective:To study the role of IL 2Rγ chain in the immuno senescence process Methods:8 young, aged from 18 to 35 years(average 24 9±5 8 years), and 17 old healthy people, aged from 65 to 79 years(average 69 4±4 2 years), had been selected for determination the proliferative responses of peripheral blood lymphocytes to PHA, the percentage and dynamic changes of CD132, CD45R positive cells, also, the gene expression of IL 2Rγ chain with RT PCR using the designed primers Results:The results showed that the PHA proliferative response of lymphocytes of aged people(0 199±0 055) were lower than that of young people(0 279±0 054), P=0 0012 The expression of IL 2Rγ chain on the lymphocytes surface decreased in aged people, 46 2%±7 7%, compared to young people, 68 0%±11 8%, P<0 001, It was verified from RT PCR result that the event occurred at transcription level The time dynamic curve of IL 2Rγ chain expression were different between aged and young people Conclusion:A preliminary conclusion would be drawn that the IL 2Rγ chain play an important role in the declined immune function of aged
