探討RANTES啟動子在三種免疫細胞內受六種不同病毒感染時所激發之表達程度

趨化激素（Chemokines）是ㄧ群小分子胜肽，由70-130個氨基酸所組成，分子量大約在6-14/kDa之間。依照其N端最後兩個胱胺酸的排列方式，可以將趨化激素分成四類，分別為C，CC，CXC，CX3C。在體外的實驗中，CC chemokines最先被證實具有抑制HIV-1（Human Immunodeficiency Virus）作用的水溶性分子。在先前我們實驗室的研究中，在台灣有發現9個人過去曾和感染HIV的人發生性行為但卻不會被感染，並且在他們的血清中可以發現高濃度的CC chemokines（MIP-1α, MIP-1β, and RANTES）。我們有興趣去了解CC chemokines成員之ㄧ的RANTES啟動子，在細胞中加或不加病毒刺激的激發程度。首先建構了不同基因型及不同刪減長度的RANTES啟動子到pGL3載體，分別送至單核球細胞、B淋巴細胞和T淋巴細胞中觀察RANTES啟動子的活性表現；另外，取全長之RANTES啟動子，分別送至細胞中再加入腺病毒、登革熱病毒、EBV、腸病毒71型、單純疱疹病毒第一型及呼吸道融合瘤病毒等病毒刺激後觀察RANTES啟動子的活性表現。結果顯示，在不同基因型中以-403G/-28C此基因型的表現高於其他基因型，於不同刪減長度之RANTES啟動子結果推測，在RANTES啟動子上可能有轉錄因子可辨認的位置，進而去調控RANTES啟動子的表現。另外，在細胞受病毒刺激後的結果顯示，當病毒感染細胞時會刺激細胞活化進而影響chemokines的表現。結果證實了細胞在遭受病毒感染後，細胞會被刺激活化並開始製造分泌chemokines，而分泌出來的chemokines是否會作用至鄰近其他細胞進而促使細胞活化，其作用機制尚未明確，在未來可以更進一步去了解及探討。

Chemokines are a group of small peptides composed of 70-130 amino acids with molecular weight of 6-14 kDa , and distinguished into four group: C,CC,CXC, and CX3C, according to their final two cystein of the amine bases. CC chemokines have been described as primary compontents of human immunodeficiency virus type 1（HIV-1） suppressive soluble factors in vitro. Previous study stated that 9 Chinese subjects in Taiwan were sexually exposed to HIV carriers and remained uninfected, these subjects found to have large quantities of CC chemokines ( MIP-1α, MIP-1β, and RANTES ) in the serum. We intresting to know that the RNATES promoter with different viruses stimulate of expression level. For this, I constructed the different genotype and serial deleted RANTES promoters to pGL3-Basic vector. First, These promoter constructs were then transfected into monocytes, B lymphocytes, and T lymphocytes to assay the RANTES promoter activity. Second, the full-length RANTES promoter were then transfected into cells to assay the activities of the RANTES promoter expression level by Adenovirus, Dengue virus, EBV, Enterovirus 71, HSV-1, and RSV infections. The first result showed that the -403G/-28C RANTES promoter genotype luciferase activity for more than other genotype. Second, the luciferase assay showed that maybe has transcription factor binding site on the RANTES promoter, then regulated of the RANTES promoter expression.Final result showed that RANTES promoter expression to increase when cell to incur viral infections. These results suggest that when viral infected cell will stimulate cell to activation then start to make up of chemokines and secretion. The mechanism of that the chemokines whether or not to affect neighbour cells activation was unknown and the further studies are required to understand the regulation mechanism.