發(fā)布時間：2018-11-17 12:20

【摘要】： 近年的研究表明,具有特定序列的單鏈脫氧寡核苷酸(ODN)具有免疫負調節(jié)作用,可能通過干預免疫系統(tǒng)或免疫細胞的異�；罨�,成為治療自身免疫病、超敏反應和Toll樣受體活化相關性疾病的藥物。這類ODN被稱為抑制性ODN,為人王合成的、長約幾十個核苷酸的單鏈DNA分子。有研究表明,抑制性ODN可以抑制含CpG基序ODN(CpG ODN)的免疫刺激作用,也可能抑制LPS誘導的免疫反應等,且多為富含鳥嘌吟(G)的ODN。 根據哺乳動物線粒體DNA的序列特點以及對可能具有免疫負調節(jié)活性ODN的結構特性認識,我們設計了13條單鏈脫氧寡核苷酸,分別命名為MT00～12。然后,利用生物學活性方法對MT00～12是否能抑制免疫激活劑誘導免疫細胞產生抗病毒物質進行了篩選,發(fā)現富含胞嘧啶的MT01表現出較強的抑制活性。進一步研究顯示:1)MT01不僅能抑制TLR9激動劑CpG ODN誘導的免疫反應,也能抑制滅活的DNA病毒、TLR7激動劑咪喹莫特或滅活的RNA病毒誘導的免疫反應,但對TLR4誘導的免疫反應則無抑制作用;2)MT01對SLE病人血清刺激的免疫反應也具有明顯的抑制作用:3)MT01的結構,尤其富含胞嘧啶區(qū)域與其生物學活性直接相關,構效關系研究顯示其免疫負調節(jié)活性可能是序列依賴性的:4)MT01在小鼠體內能明顯抑制CpG ODN對乙肝表面抗原特異性抗體誘生的增強作用,但對乙肝表面抗原刺激抗體產生無抑制作用,說明MT01可能只對過強的免疫反應具有抑制作用;5)MT01可抑制小鼠B細胞活化,這提示其對抗原特異性抗體產生的抑制作用可能是通過抑制B細胞活化而實現的。 綜上,一種富含胞嘧啶的單鏈脫氧寡核苷酸(MT01)具有選擇性免疫負調節(jié)活性,可能具有治療自身免疫病或TLR7/9活化相關性疾病的潛在應用價值。
[Abstract]:Recent studies have shown that single-stranded oligonucleotides (ODN) with specific sequences have negative immunomodulatory effects, which may be used to treat autoimmune diseases by interfering with the abnormal activation of immune system or immune cells. Drugs associated with hypersensitivity and Toll-like receptor activation-associated diseases. This type of ODN is known as a mono-stranded DNA molecule of about dozens of nucleotides synthesized by inhibitory ODN,. Some studies have shown that inhibitory ODN can inhibit the immunostimulatory effect of ODN (CpG ODN) containing CpG motif, and may also inhibit the immune response induced by LPS, and most of them are ODN. rich in CpG (G). According to the sequence characteristics of mammalian mitochondrial DNA and the recognition of the structural characteristics of ODN with possible immuno-negative regulatory activity, we designed 13 single-stranded oligodeoxynucleotides named MT00~12.. Then, the biological activity method was used to screen whether MT00~12 could inhibit the production of antiviral substances in immune cells induced by immunoactivator. It was found that MT01 rich in cytosine showed strong inhibitory activity. Further studies showed that: 1) MT01 could not only inhibit the immune response induced by TLR9 agonist CpG ODN, but also inhibit the immune response induced by inactivated DNA virus, TLR7 agonist Imiquimod or inactivated RNA virus. But the immune response induced by TLR4 was not inhibited. 2) MT01 also inhibited the immune response to serum stimulation in patients with SLE. 3) the structure of MT01, especially cytosine rich region, was directly related to its biological activity. The study of structure-activity relationship showed that its negative immunomodulatory activity might be sequence-dependent: 4) MT01 could significantly inhibit the enhancement of CpG ODN induced by HBsAg specific antibody in mice. However, there was no inhibitory effect on the antibody stimulated by hepatitis B surface antigen, indicating that MT01 may only inhibit the excessive immune response. 5) MT01 can inhibit the activation of B cells in mice, which suggests that the inhibition of antigen-specific antibodies may be achieved by inhibiting the activation of B cells. In conclusion, a cytosine rich single-strand deoxyoligodeoxynucleotide (MT01) has selective negative immunomodulatory activity, which may have potential application value in the treatment of autoimmune diseases or TLR7/9 activation-related diseases.
【學位授予單位】：吉林大學
【學位級別】：博士
【學位授予年份】：2009
【分類號】：R392.11

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