Characterization of bovine homologues of granulysin and NK-lysin
Characterization of bovine homologues of granulysin and NK-lysin
复制标题
DOI：
10.4049/jimmunol.173.4.2607
复制
发表时间：
2004-08-15
影响因子：
4.4
通讯作者：
Estes, DM
中科院分区：
文献类型：
--
作者：
Endsley, JJ;Furrer, JL;Estes, DM
颗粒溶素和NK-溶素是在人和猪细胞毒性淋巴细胞的颗粒中发现的抗微生物蛋白。迄今为止，尚未确定颗粒溶素的鼠对应物，这表明了额外模型的重要性，以充分表征颗粒溶素样分子在对感染性疾病的免疫应答中的作用。从牛PBMC mRNA中扩增出颗粒溶素和NK-溶素的两个部分核苷酸序列，分别对应于颗粒溶素和NK-溶素的完整功能域。用佛波醇酯和钙离子载体刺激后，在CD 3（+）T细胞、CD 4（+）T细胞、CD 8（+）T细胞、WC 1（+）γ δ T细胞和去除CD 3（+）T细胞的PBMC中检测到牛基因的表达，但在CD 21（+）细胞和CD 14（+）细胞中不存在。细胞内流式细胞术和免疫印迹证实了激活的T淋巴细胞和PBMC中存在对应于牛颗粒溶素同系物的蛋白质。合成的人、牛和猪肽对应于颗粒溶素的螺旋2至螺旋3区域的C末端，对大肠杆菌、沙门氏菌、金黄色葡萄球菌和牛分枝杆菌卡介苗显示出有效的抗微生物活性。对应于螺旋2的人和牛肽显示抗分枝杆菌活性。牛卡介苗牛基因的表达在激光显微镜下解剖的淋巴结病变从M。牛感染的动物。一个生物活性的牛同源颗粒溶素的鉴定证明了牛模型的潜力，在表征颗粒溶素的免疫反应中的作用，以各种传染性病原体。
Granulysin and NK-lysin are antimicrobial proteins found in the granules of human and swine cytotoxic lymphocytes. A murine-counterpart to granulysin has not been identified to date, indicating the importance of additional models to fully characterize the role of granulysin-like molecules in the immune response to infectious disease. Two partial nucleotide sequences corresponding to the complete functional domain of granulysin and NK-lysin were amplified from bovine PBMC mRNA. Following stimulation with phorbol ester and calcium, ionophore, expression of the bovine gene was detected in CD3(+) T cells, CD4(+) T cells, CD8(+) T cells, WC1(+) gammadelta T cells, and PBMC depleted of CD3(+) T cells, but was absent in CD21(+) cells and CD14(+) cells. Intracellular How cytometry and immunoblotting confirmed the presence of protein corresponding to the bovine granulysin homologue in activated T lymphocytes and PBMC. Synthetic human, bovine, and swine peptides corresponding to the C terminus of helix 2 through helix 3 region of granulysin displayed potent antimicrobial activity against Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, and Mycobacterium bovis bacillus Calmette-Guerin. Human and bovine peptides corresponding to helix 2 displayed antimycobacterial activity against M.. bovis bacillus Calmette-Guerin. Expression of the bovine gene was detected in laser microscopy-dissected lymph node lesions from an M. bovis-infected animal. The identification of a biologically active bovine homologue to granulysin demonstrates the potential of the bovine model in characterizing the role of granulysin in the immune response to a variety of infectious agents.