局部应用鞘氨醇-1-磷酸受体调节剂抑制小鼠异基因角膜移植排斥的研究
发布时间：2019-05-16 03:36
【摘要】：目的：通过将鞘氨醇-1-磷酸(S1P)受体调节剂FTY720眼用凝胶应用于小鼠异基因角膜移植模型中，观察FTY720眼用凝胶抑制小鼠角膜移植排斥的情况，并与临床常用的眼用免疫抑制剂环孢霉素A（CsA）进行比较。最终明确FTY720眼用凝胶抑制小鼠角膜移植的排斥反应的效果。通过流式细胞学、Realtime-PCR、Elisa、常规病理以及免疫组化方法对不同处理组小鼠的角膜、外周血、脾脏及颈部淋巴结标本进行检测；以探索FTY720眼用凝胶局部点眼抑制小鼠异基因角膜移植排斥的作用机制。使用磁珠分选法分选出具有免疫调节功能的小鼠调节性T细胞，并将分选细胞分别与S1P受体调节剂（FTY720及FTY720-P）共培养。通过CCK-8法检测共培养的调节性T细胞增殖情况，以及混合淋巴反应结果；以确定S1P受体调节剂直接作用于调节性T细胞后的细胞增殖及免疫功能变化情况。通过Elisa法检测共培养体系中与细胞功能相关的细胞因子(IL-10和TGF-β1)水平；Realtime-PCR法了解共培养细胞中相应细胞因子mRNA的表达情况。最终以探索出S1P受体调剂直接作用于小鼠调节性T细胞后，其功能及表型发生变化的的机制。 方法： 1.制备0.1%、0.3%及0.5%三个浓度S1P受体调节剂FTY720温度敏感型眼用凝胶。 2.取45雄性BALB/C小鼠随机分为5组（空白对照组、低浓度、中浓度、高浓度FTY720眼用凝胶组以及1%环孢霉素眼液组），每只小鼠随机选取一眼作为受体眼，分别接受雄性C57BL/6小鼠角膜植片，进行异基因穿透性角膜移植。术后即分别给予相应方案处置，术后10-11天拆除角膜缝线；一月内显微镜下观察并记录全部小鼠的角膜植片情况。至小鼠角膜发生排斥及观察结束后处死实验动物。 3.取30只雄性BALB/C小鼠同上随机分为5组，每只小鼠随机一眼作为受体眼，分别接受雄性C57BL/6小鼠角膜植片，进行异基因穿透性角膜移植。术后分别给予上述方案分组方案处置，术后10-11天拆除角膜缝线。术后14天时处死小鼠，分别进行实验室检查。其中取颈部引流淋巴结、外周血、脾脏进行流式细胞学检查，了解CD4+T细胞及CD4+CD25+Foxp3+T细胞的分布情况。对外周血的标本使用Elisa法检测外周血清中IL-2、IL-10、IFN-γ及TGF-β1的含量。每组取3只小鼠角膜分别采用RealtimePCR法检测角膜植片中IL-2、IL-10、IFN-γ、TGF-β1及Foxp3mRNA表达情况。每组另取3只小鼠角膜分别进行常规HE染色，了解角膜排斥反应情况；并对CD4+T细胞以及细胞因子IL-2、IL-10、IFN-γ及TGF-β1进行免疫组化检查，了解CD4+T细胞在角膜植片浸润情况以及上述有关细胞因子在角膜植片中含量的变化。 4.取30只雄性BALB/C小鼠；处死后应用免疫磁珠分选法将脾脏中的CD4+CD25+以及CD4+CD25-T细胞分选出来。对分选后的细胞采用流式细胞技术明确分选效率。 5.对分选的细胞应用CD3、CD28单抗联合IL-2共同体外培养5天，充分激活上述分选细胞。 6.在激活的CD4+CD25+T细胞培养体系中分别加入10ng/ml、100ng/ml、1000ng/ml的FTY720以及FTY720-P，空白对照组加入10ulDMSO；继续共培养48小时。CCK-8法检测CD4+CD25+T细胞加入S1P受体调节剂前后细胞增殖情况（共计5天，第0天为加入S1P受体调节剂时、第1天及2天为加入S1P受体调节剂后时刻，第3及4天为洗涤上述细胞后继续原培养条件进行培养的时刻）。 7.加入不同药物后的48小时后，取共培养后的CD4+CD25+T细胞（调节性T细胞、Treg）与CD4+CD25-T细胞（效应T细胞、Teff）按不同比例进行混合淋巴细胞培养，5天后CCK-8法检测Teff细胞增殖情况。 8.加入不同药物后的48小时后，取共培养CD4+CD25+T细胞上清液进行Elisa检测，检测IL-10、TGF-β1含量；从上述共培养细胞中提取总RNA，Realtime-PCR检测IL-10、TGF-β1及Foxp3mRNA的表达情况。 结果： 1.成功制备0.1%、0.3%及0.5%三个浓度S1P受体调节剂FTY720温度敏感型眼用凝胶。其在室温环境下为液态，在近体温状态时迅速变为胶冻状。 2.观察45只小鼠术后角膜植片情况，植片平均生存时间（MST）方面，0.5%FTY720组(24.11±1.58天)以及1%CsA组(25.00±1.91天)均较空白对照组（13.44±0.48天）明显延长(P0.01)。 3.0.5%FTY720能显著增加颈部淋巴结中CD4+T细胞以及Treg细胞的比例(P0.05和P0.01)。在0.5%FTY720组，角膜植片中TGF-β1mRNA表达明显增高(p0.05),而在1%CsA组角膜植片中IL-2和IFN-γmRNA则明显降低的(p0.01和p0.05)。免疫组化染色结果证实角膜植片中上述细胞因子含量变化情况与相应mRNA表达基本一致。此外0.5%FTY720以及1%CsA点眼后，角膜植片中CD4+T细胞的浸润明显减少。 4.全部各组小鼠外周血清中IL-2、IL-10、IFN-γ及TGF-β1含量未见变化。 5.自小鼠脾脏细胞中成功分选CD4+CD25+T细胞，流式细胞检测结果显示CD4+CD25+T细胞的分选效率在93%以上，分选出细胞中Foxp3+比例在94%以上。CD4+CD25-T细胞分选率接近96%其Foxp3+比例仅为1.45%左右。 6.对上述分选出的细胞应用CD3、CD28单抗联合IL-2共同体外培养5天后，上述细胞被充分激活。 7.对激活的的CD4+CD25+T细胞分别给予三个不同剂量的FTY720及FTY720-P共培养后，，经过CCK-8检测后发现与空白对照组比较，给予S1P受体调节剂后CD4+CD25+T细胞的增殖活性未见变化（P0.05）。 8.共培养的Treg细胞与Teff细胞进行混合淋巴细胞培养5天后，结果显示在Treg:Teff为1:1时，高剂量FTY720、中等剂量及高剂量的FTY720-P组中Teff细胞增殖能力被明显抑制（P0.05）；在Treg:Teff为1:4时，仅有高剂量的FTY720-P组中Teff细胞增殖能力被抑制（P0.05）；而在Treg:Teff为1:8时，各组之间未见显著性差异（P0.05）。 9.共培养48小时后，取共培养CD4+CD25+T细胞上清液进行Elisa检测。结果显示检测各组中IL-10含量未见变化；对于TGF-β1含量，在高剂量FTY720组、中等剂量和高剂量FTY720-P组的含量均显著高于对照组（P0.05）。对上述共培养细胞进行RealtimePCR检测，结果显示三个剂量FTY720组细胞中IL-10、TGF-β1及Foxp3mRNA水平未见显著变化；而在高剂量FTY720-P组细胞中TGF-β1及Foxp3mRNA的表达明显增高（P0.05）。 结论： 1.0.5%FTY720眼用凝胶以及1%CsA眼液均能够显著延长异基因小鼠角膜移植术后免疫排斥反应的发生时间。 2.0.5%FTY720眼用凝胶能够增加颈部引流淋巴结中的CD4+T细胞的比率，且更为显著地增加了Treg细胞在局部淋巴结中的比率。0.5%FTY720能够增加角膜植片中TGF-β1mRNA及其蛋白的表达；并且能够减少CD4+T细胞在角膜植片中的浸润。 3.1%CsA眼液也能够显著减少CD4+T细胞在角膜植片中的浸润；但是与0.5%FTY720眼用凝胶不同，其角膜植片中IL-2及IFN-γ的含量显著减少。 4.各组血清中有关细胞因子的含量未见变化（局部点眼剂型未能影响到外周血中细胞因子的含量）。 5.三个浓度的FTY720及FTY720-P均不能影响共培养Treg细胞的增殖能力。 6.高剂量FTY720、中等及高剂量的FTY720-P能够显著增加共培养Treg细胞针对Teff细胞的抑制能力。其作用机制为增加了共培养细胞中CD25+及Foxp3+表型的比例，并且上调了共培养细胞中TGF-β1mRNA及相应蛋白的表达；此外高剂量FTY720-P也能够增加共培养Treg细胞中Foxp3mRNA表达。
[Abstract]:Objective: To observe the effect of FTY720 ophthalmic gel on corneal graft rejection in mouse and compare with the commonly used ocular immunosuppressant cyclosporin A (CsA). Finally, the effect of FTY720 ophthalmic gel on the rejection of corneal transplantation in mice was determined. The corneal, peripheral blood, spleen and neck lymph node specimens of different treatment group mice were tested by flow cytometry, Realtime-PCR, Elisa, routine pathology and immunohistochemistry. The regulatory T cells with immunomodulatory function were selected by magnetic bead sorting and the cells were co-cultured with S1P receptor modulators (FTY720 and FTY720-P), respectively. The proliferation of the co-cultured regulatory T cells was detected by the CCK-8 method, and the results of the mixed lymph reaction were determined to determine the cell proliferation and the change of the immune function of the S1P receptor modulator directly after the regulatory T cells. The level of cytokines (IL-10 and TGF-1 1) related to cell function in co-culture system was detected by Elisa method, and the expression of corresponding cytokine mRNA in co-cultured cells was obtained by Realtime-PCR. Finally, the mechanism of the change of the function and phenotype of the S1P receptor after the regulatory T cell of the mouse was explored. square Method:1. The temperature-sensitive type of FTY720, which is 0.1%, 0.3% and 0.5%, has three concentrations of S1P receptor modulator, FTY720, is prepared. 2.45 male BALB/ C mice were randomly divided into 5 groups (blank control group, low concentration, medium concentration, high-concentration FTY720 ophthalmic gel group and 1% cyclosporin eye group), and each mouse was randomly selected as the receptor eye, and the male C57BL/6 mice were respectively accepted. mouse corneal graft The corresponding treatment was given after operation, and the corneal suture was removed for 10-11 days post-operation, and the angle of all mice was observed and recorded in a 1-month internal microscope. in that case of membrane-graft, at the end of the rejection of the cornea of the mouse and the end of the observation 3.30 male BALB/ C mice were randomly divided into 5 groups, and each mouse was given a random glance as the receptor eye to receive the male C57BL/6 mouse corneal graft and to carry out the isogenic gene. Penetrating keratoplasty, the above-mentioned protocol group protocol was given after operation, and the procedure was 10-11 after operation. The corneal suture was removed on day. The mice were sacrificed at 14 days post-operation, respectively Conduct the laboratory test. The lymph node, peripheral blood and spleen of the neck were taken for flow cytometry to understand the CD4 + T cells and the CD4 + CD25 + Foxp3 + T. The distribution of IL-2, IL-10, IFN-1 and TG in peripheral blood were detected by Elisa method. The levels of IL-2, IL-10, IFN-1, TGF-CD1 and Foxp3m were detected by using the RealtimePCR method. In each group, three mouse corneas were used for routine HE staining to understand the corneal rejection, and CD4 + T cells as well as the cytokines IL-2, IL-10, IFN-1 and TGF-CD1 were fed into each group. Immunohistochemical examination was performed to understand the infiltration of CD4 + T cells in the cornea and the related cytokines in the cornea. Changes in the content of the tablets.4.30 male BALB/ C mice were taken; the CD4 + CD25 + and CD4 + CD2 in the spleen were treated with the immunomagnetic bead sorting method after termination 5-T cells are sorted out. 5. The cells were treated with CD3, CD28 and IL-2, and cultured for 5 days. 6. In the activated CD4 + CD25 + T cell culture system,10 ng/ ml,100 ng/ ml,1000 ng/ ml of FTY720 and FTY720-P were added, respectively, and the blank control group was added with 10 ulD. MSO; continued co-culture for 48 h. CCK-8 method for the detection of the proliferation of CD4 + CD25 + T cells before and after the addition of S1P receptor modulators (total 5 days, day 0, with S1P receptor modulator, day 1 and 2) after the S1P receptor modulator is added, the cells are washed after the cells are washed in days 3 and 4 7. After 48 hours after the addition of different drugs, the co-cultured CD4 + CD25 + T cells (regulatory T cells, Tregs) and the CD4 + CD25-T cells (effector T cells, Teff) were mixed for lymphocyte culture in different proportions, and CCK was used after 5 days. 8. After 48 hours after the addition of different drugs, the supernatant of the co-cultured CD4 + CD25 + T cells was taken for Elisa test to detect the content of IL-10 and TGF-CD1; total RNA was extracted from the co-cultured cells, and the real time-PCR was used to detect the IL-10 and TGF- the first and the second Results:1. The expression of Foxp3mRNA was 0.1%, 0.3% and 0.5%, respectively. P receptor modulator FTY720 temperature sensitive ophthalmic gel at room temperature In the case of 45 mice, the average survival time (MST), 0.5% FTY720 group (24.11-1.58 days) and 1% CsA (25.00-1.91 days) were compared with the blank control group (25.00-1.91 days). 13.44 (0.48 days) significantly prolonged (P0.01). 3.0.5% FTY720 significantly increased the CD4 + T fine in the cervical lymph node In the 0.5% FTY720 group, the expression of TGF-CD1 mRNA in the corneal grafts was significantly higher (p0.05), while IL-2 and IFN in the 1% CsA group were significantly higher (p0.05). -The results of the immunohistochemical staining showed that the expression of the mRNA in the cornea was significantly lower (p0.01 and p0.05). in addition, 0.5% FTY720 and 1% C 4. There was a significant decrease in the infiltration of CD4 + T cells in the corneal grafts after the eyes of the sA. 5. CD4 + CD25 + T cells were successfully sorted from the spleen cells of mice. The results of flow cytometry showed that the CD4 + CD25 + T cells were sorted. The efficiency is over 93%, and the ratio of Foxp3 + in the selected cells is over 94%. CD4 + CD25- The cell sorting rate of T cells was close to 96%, and the ratio of Foxp3 + was only about 1.45%. After three different doses of FTY720 and FTY720-P, the activated CD4 + CD25 + T cells were co-cultured with three different doses of FTY720 and FTY720-P. The proliferation activity of CD4 + CD25 + T cells was not changed after body conditioning (P0.05).8. The co-cultured Treg cells were mixed with Teff cells for 5 days, and the results showed that Tef in the high dose of FTY720, medium dose and high dose of FTY720-P group at 1:1 in Treg: Teff The proliferation ability of the f cells was significantly inhibited (P0.05); in the case of Treg: Teff of 1:4, the proliferation ability of Teff cells in the FTY720-P group with only high dose was inhibited (P0.05); However, there was no significant difference between the groups at the time of Treg: Teff (P0.05). After the co-culture for 48 hours, the supernatant of the co-cultured CD4 + CD25 + T cells was taken for Elisa detection. The results showed no change in the content of IL-10 in each group; for the TGF-1 content, at high dose of FTY720 The results showed that the levels of IL-10, TGF-CD1 and Foxp3mRNA in the three-dose FTY720 group did not change significantly in the group, middle dose and high-dose FTY720-P group, while at the high dose of FTY72, the levels of IL-10, TGF-1 and Foxp3mRNA in the three-dose FTY720 group were not significantly changed. 0-P The expression of TGF-CD1 and Foxp3mRNA in the group was significantly higher (P0.05). Conclusion:1.5% FTY720 eyes 2.0.5% of FTY720 ophthalmic gel can increase the ratio of CD4 + T cells in the lymph nodes of the neck, and increase the ratio of the Treg cells to the local lymph nodes more significantly. 0.5% of the FTY720 can increase the cornea. The expression of TGF-CD1 mRNA and its protein in the implanted tablet can be reduced, and the infiltration of CD4 + T cells in the corneal implant can be reduced. 3.1% CsA can also significantly reduce the infiltration of CD4 + T cells in the corneal implant. The results showed that the content of IL-2 and IFN-1 in the corneal implant was significantly lower than that of the 0.5% FTY720 ophthalmic gel. .4. There was no change in the content of the relevant cytokines in each group (the local point-to-eye dosage form failed to affect the cells in the peripheral blood). 5. The three concentrations of FTY720 and FTY720-P could not affect the proliferation of the co-cultured Treg cells. The amount of FTY720, medium and high doses of FTY720-P can significantly increase the inhibitory capacity of co-cultured Treg cells against Teff cells. The mechanism of action is to increase the ratio of CD25 + and Foxp3 + phenotype in co-cultured cells and up-regulate the TGF-+ 1 m in co-cultured cells.
【学位授予单位】：中国人民解放军军医进修学院
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R779.65
本文编号：2477986
[Abstract]:Objective: To observe the effect of FTY720 ophthalmic gel on corneal graft rejection in mouse and compare with the commonly used ocular immunosuppressant cyclosporin A (CsA). Finally, the effect of FTY720 ophthalmic gel on the rejection of corneal transplantation in mice was determined. The corneal, peripheral blood, spleen and neck lymph node specimens of different treatment group mice were tested by flow cytometry, Realtime-PCR, Elisa, routine pathology and immunohistochemistry. The regulatory T cells with immunomodulatory function were selected by magnetic bead sorting and the cells were co-cultured with S1P receptor modulators (FTY720 and FTY720-P), respectively. The proliferation of the co-cultured regulatory T cells was detected by the CCK-8 method, and the results of the mixed lymph reaction were determined to determine the cell proliferation and the change of the immune function of the S1P receptor modulator directly after the regulatory T cells. The level of cytokines (IL-10 and TGF-1 1) related to cell function in co-culture system was detected by Elisa method, and the expression of corresponding cytokine mRNA in co-cultured cells was obtained by Realtime-PCR. Finally, the mechanism of the change of the function and phenotype of the S1P receptor after the regulatory T cell of the mouse was explored. square Method:1. The temperature-sensitive type of FTY720, which is 0.1%, 0.3% and 0.5%, has three concentrations of S1P receptor modulator, FTY720, is prepared. 2.45 male BALB/ C mice were randomly divided into 5 groups (blank control group, low concentration, medium concentration, high-concentration FTY720 ophthalmic gel group and 1% cyclosporin eye group), and each mouse was randomly selected as the receptor eye, and the male C57BL/6 mice were respectively accepted. mouse corneal graft The corresponding treatment was given after operation, and the corneal suture was removed for 10-11 days post-operation, and the angle of all mice was observed and recorded in a 1-month internal microscope. in that case of membrane-graft, at the end of the rejection of the cornea of the mouse and the end of the observation 3.30 male BALB/ C mice were randomly divided into 5 groups, and each mouse was given a random glance as the receptor eye to receive the male C57BL/6 mouse corneal graft and to carry out the isogenic gene. Penetrating keratoplasty, the above-mentioned protocol group protocol was given after operation, and the procedure was 10-11 after operation. The corneal suture was removed on day. The mice were sacrificed at 14 days post-operation, respectively Conduct the laboratory test. The lymph node, peripheral blood and spleen of the neck were taken for flow cytometry to understand the CD4 + T cells and the CD4 + CD25 + Foxp3 + T. The distribution of IL-2, IL-10, IFN-1 and TG in peripheral blood were detected by Elisa method. The levels of IL-2, IL-10, IFN-1, TGF-CD1 and Foxp3m were detected by using the RealtimePCR method. In each group, three mouse corneas were used for routine HE staining to understand the corneal rejection, and CD4 + T cells as well as the cytokines IL-2, IL-10, IFN-1 and TGF-CD1 were fed into each group. Immunohistochemical examination was performed to understand the infiltration of CD4 + T cells in the cornea and the related cytokines in the cornea. Changes in the content of the tablets.4.30 male BALB/ C mice were taken; the CD4 + CD25 + and CD4 + CD2 in the spleen were treated with the immunomagnetic bead sorting method after termination 5-T cells are sorted out. 5. The cells were treated with CD3, CD28 and IL-2, and cultured for 5 days. 6. In the activated CD4 + CD25 + T cell culture system,10 ng/ ml,100 ng/ ml,1000 ng/ ml of FTY720 and FTY720-P were added, respectively, and the blank control group was added with 10 ulD. MSO; continued co-culture for 48 h. CCK-8 method for the detection of the proliferation of CD4 + CD25 + T cells before and after the addition of S1P receptor modulators (total 5 days, day 0, with S1P receptor modulator, day 1 and 2) after the S1P receptor modulator is added, the cells are washed after the cells are washed in days 3 and 4 7. After 48 hours after the addition of different drugs, the co-cultured CD4 + CD25 + T cells (regulatory T cells, Tregs) and the CD4 + CD25-T cells (effector T cells, Teff) were mixed for lymphocyte culture in different proportions, and CCK was used after 5 days. 8. After 48 hours after the addition of different drugs, the supernatant of the co-cultured CD4 + CD25 + T cells was taken for Elisa test to detect the content of IL-10 and TGF-CD1; total RNA was extracted from the co-cultured cells, and the real time-PCR was used to detect the IL-10 and TGF- the first and the second Results:1. The expression of Foxp3mRNA was 0.1%, 0.3% and 0.5%, respectively. P receptor modulator FTY720 temperature sensitive ophthalmic gel at room temperature In the case of 45 mice, the average survival time (MST), 0.5% FTY720 group (24.11-1.58 days) and 1% CsA (25.00-1.91 days) were compared with the blank control group (25.00-1.91 days). 13.44 (0.48 days) significantly prolonged (P0.01). 3.0.5% FTY720 significantly increased the CD4 + T fine in the cervical lymph node In the 0.5% FTY720 group, the expression of TGF-CD1 mRNA in the corneal grafts was significantly higher (p0.05), while IL-2 and IFN in the 1% CsA group were significantly higher (p0.05). -The results of the immunohistochemical staining showed that the expression of the mRNA in the cornea was significantly lower (p0.01 and p0.05). in addition, 0.5% FTY720 and 1% C 4. There was a significant decrease in the infiltration of CD4 + T cells in the corneal grafts after the eyes of the sA. 5. CD4 + CD25 + T cells were successfully sorted from the spleen cells of mice. The results of flow cytometry showed that the CD4 + CD25 + T cells were sorted. The efficiency is over 93%, and the ratio of Foxp3 + in the selected cells is over 94%. CD4 + CD25- The cell sorting rate of T cells was close to 96%, and the ratio of Foxp3 + was only about 1.45%. After three different doses of FTY720 and FTY720-P, the activated CD4 + CD25 + T cells were co-cultured with three different doses of FTY720 and FTY720-P. The proliferation activity of CD4 + CD25 + T cells was not changed after body conditioning (P0.05).8. The co-cultured Treg cells were mixed with Teff cells for 5 days, and the results showed that Tef in the high dose of FTY720, medium dose and high dose of FTY720-P group at 1:1 in Treg: Teff The proliferation ability of the f cells was significantly inhibited (P0.05); in the case of Treg: Teff of 1:4, the proliferation ability of Teff cells in the FTY720-P group with only high dose was inhibited (P0.05); However, there was no significant difference between the groups at the time of Treg: Teff (P0.05). After the co-culture for 48 hours, the supernatant of the co-cultured CD4 + CD25 + T cells was taken for Elisa detection. The results showed no change in the content of IL-10 in each group; for the TGF-1 content, at high dose of FTY720 The results showed that the levels of IL-10, TGF-CD1 and Foxp3mRNA in the three-dose FTY720 group did not change significantly in the group, middle dose and high-dose FTY720-P group, while at the high dose of FTY72, the levels of IL-10, TGF-1 and Foxp3mRNA in the three-dose FTY720 group were not significantly changed. 0-P The expression of TGF-CD1 and Foxp3mRNA in the group was significantly higher (P0.05). Conclusion:1.5% FTY720 eyes 2.0.5% of FTY720 ophthalmic gel can increase the ratio of CD4 + T cells in the lymph nodes of the neck, and increase the ratio of the Treg cells to the local lymph nodes more significantly. 0.5% of the FTY720 can increase the cornea. The expression of TGF-CD1 mRNA and its protein in the implanted tablet can be reduced, and the infiltration of CD4 + T cells in the corneal implant can be reduced. 3.1% CsA can also significantly reduce the infiltration of CD4 + T cells in the corneal implant. The results showed that the content of IL-2 and IFN-1 in the corneal implant was significantly lower than that of the 0.5% FTY720 ophthalmic gel. .4. There was no change in the content of the relevant cytokines in each group (the local point-to-eye dosage form failed to affect the cells in the peripheral blood). 5. The three concentrations of FTY720 and FTY720-P could not affect the proliferation of the co-cultured Treg cells. The amount of FTY720, medium and high doses of FTY720-P can significantly increase the inhibitory capacity of co-cultured Treg cells against Teff cells. The mechanism of action is to increase the ratio of CD25 + and Foxp3 + phenotype in co-cultured cells and up-regulate the TGF-+ 1 m in co-cultured cells.
【学位授予单位】：中国人民解放军军医进修学院
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R779.65
【共引文献】
相关硕士学位论文 前1条
1 金卉;大肠腺癌中鞘氨醇激酶-1与1-磷酸鞘氨醇受体mRNA和蛋白表达的研究[D];广西医科大学;2010年
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