日本血吸虫可溶性虫卵抗原免疫小鼠DC亚群对哮喘抑制作用的实验研究
发布时间：2018-09-08 20:15
【摘要】：目的通过过继转移日本血吸虫可溶性虫卵抗原(SEA)免疫小鼠树突状细胞(DC)亚群,探讨SEA免疫的DC亚群在抑制过敏性哮喘中的作用及可能的作用机制。 方法BALB/c小鼠经腹腔及足垫注射SEA50μg/只,每周1次,共4次。对照组注射生理盐水。用抗体包被的免疫磁珠分离小鼠脾CD11c+CD8α+DC与CD11c+CD8α-DC亚群。另取30只BALB/c小鼠,随机分为6组,A组为正常对照组(SCN),B组为单纯哮喘组(SNO),C组为过继转移盐水对照CD11c+CD8α+DC组(AA), D组为过继转移盐水对照CD11c+CD8α-DC组(AB),E组为过继转移SEA免疫CD11c+CD8α+DC组(SA),F组为过继转移SEA免疫CD11c+CD8α-DC组(SB)。C、D组小鼠分别经尾静脉过继转移5×105个盐水对照CD11c+CD8α+DC和'5x105个盐水对照CD11c+CD8α-DC; E、F组小鼠分别经尾静脉过继转移SEA免疫5×105个CD11c+CD8α+DC和5×105个SEA免疫CD11c+CD8α-TC。1h后B、C、D、E、F组小鼠同时用卵白蛋白(OVA)诱发哮喘,4周后剖杀,取肺组织,做病理切片,经苏木精-伊红(HE)染色,光镜下观察肺组织炎症变化；收集小鼠支气管肺泡灌洗液(BALF)进行嗜酸性粒细胞(EOS)计数及细胞因子IL-4、IL-5、IL-10、IL-12、转化生长因子β(TGF-β)检测；收集血清检测OVA特异性IgE抗体水平；收集脾细胞培养上清(SPC)进行细胞因子IL-4、IL-5、IL-10、IL-12、TGF-β检测。结果肺组织病理切片HE染色结果显示,与B、C、D、E组相比,F组小鼠肺组织炎症明显减轻,肺组织病理学总评分有统计学意义(P0.05)；A、B、C、D、E、F六组小鼠BALF中EOS计数百分比分别为(8.40±1.14)、(40.00±3.39)、(23.67±4.62)、(25.20±3.19)、(20.33±3.22)和(13.00±1.58),与B、C、D、E组比较,F组EOS数目明显减少(P0.05)；A、B、C、D、E、F六组小鼠血清OVA特异性IgE水平分别为(204.90±3.58)pg/ml、(1127.83±18.86)pg/ml、(1097.03±25.80)pg/ml、(1097.08±17.23)pg/ml、(1039.78±2.90)pg/ml、(319.33±3.97)pg/ml,与B、C、D、E组比较,F组IgE水平显著下降(P0.05); A、B、C、D、E、F六组小鼠BALF中IL-4分泌水平分别为(10.17±4.49)pg/ml、(80.36±10.94)pg/ml、(67.83±6.81)pg/ml、(62.41±4.02)pg/ml、(57.33±11.82)pg/ml、(22.36±7.62)pg/ml、IL-5分泌水平分别为(17.13±7.33)pg/ml、(1471.24±56.15)pg/ml、(823.37±6.52)pg/ml、(833.18±35.46)pg/ml、(811.16±11.61)pg/ml、(313.77±2.97)pg/ml; IL-10分泌水平分别为(120.79±21.48)pg/ml、(72.77±17.60)pg/ml、(81.63±9.08)pg/ml、(82.80±2394)pg/ml、(106.94±26.31)pg/ml、(248.89±33.20)pg/ml; IL-12分泌水平分别为(50.37±1.53)pg/ml,(46.70±7.00)pg/ml、(122.37±16.01)pg/ml、(136.45±9.67)pg/ml、(140.53±4.81)pg/ml、(340.95±22.22)pg/ml; TGF-P分泌水平分别为(176.91±51.25)pg/ml、(190.09±20.64)pg/mk (109.64±34.71)pg/ml、(162.27±38.95)pg/ml、(220.09±25.00)pg/ml、(966.82±57.48)pg/ml、与B、C、D、E组比较,F组IL-4、IL-5水平明显降低(P0.05), IL-10、IL-12、TGF-β水平显著升高(P0.05)。A、B、C、D、E、F六组小鼠SPC中IL-4分泌水平分别为(17.77±7.66)pg/ml、(248.77±18.85)pg/ml、(220.51±11.84)pg/ml、(217.13±50.37)pg/ml、(172.70±27.81)pg/ml、(76.90±14.92)pg/m;IL-5分泌水平分别为(12.13±2.96)pg/ml、(1770.84±27.50)pg/ml、(1082.34±78.44)pg/ml、(1124.81±22.62)pg/ml、(1058.96±38.68)pg/ml、(451.16±15.72)pg/ml; IL-10分泌水平分别为(122.88±16.81)pg/ml、(131.10±11.52)pg/ml、(161.21±1.80)pg/ml、(204.91±8.71)pg/ml、(220.90±26.86)pg/ml、(998.81±138.46)pg/ml; IL-12分泌水平分别为(14.03±5.13)pg/ml、(66.03+2.31)pg/ml、(75.03±14.01)pg/ml、(132.20±22.69)pg/ml、(179.53±16.74)pg/mk (405.70±57.66)pg/ml; TGF-β分泌水平分别为(128.05l±14.79)pg/ml、(102.14±46.03)pg/ml、(168.27±60.69)pg/ml、(183.27±79.13)pg/ml、(160.55±29.64)pg/ml、(1101.55±73.60)pg/ml;与B、C、D、E组比较,F组IL-4、IL-5水平明显降低(P0.05), IL-10、IL-12、TGF-β水平显著升高(P0.05)。结论日本血吸虫SEA免疫的DC亚群对过敏性哮喘有抑制作用,该抑制作用主要通过CD8α-DC亚群产生。
[Abstract]:Objective To investigate the role and possible mechanism of DC subpopulation immunized with SEA in inhibiting allergic asthma by adoptive transfer of soluble egg antigen (SEA) from Schistosoma japonicum.
Methods BALB/c mice were injected with SEA 50 ug/mouse via abdominal cavity and footpad once a week for 4 times.The control group was injected with normal saline.The spleen CD11c+CD8a+DC and CD11c+CD8a-DC subgroups were separated by immunomagnetic beads coated with antibody.Another 30 BALB/c mice were randomly divided into 6 groups.Group A was normal control group(SCN),group B was simple asthma group(SNO),and group C was adoptive transfer. CD11c+CD8a+DC group (AA), CD11c+CD8a-DC group (AB), SEA-immunized CD11c+CD8a+DC group (SA), SEA-immunized CD11c+CD8a+DC group (SB). C. D group mice were subjected to adoptive transfer of CD11c+CD8a+DC and'5x105 saline control CD11c+C+CD8a+DC via caudal vein, respectively. D8a-DC; E, F mice were immunized with 5 *105 CD11c+CD8a+DC and 5 *105 SEA-immunized CD11c+CD8a-TC respectively by tail vein adoptive transfer SEA. At the same time, B, C, D, E, F mice were immunized with ovalbumin (OVA) to induce asthma. Four weeks later, the lung tissues were dissected and pathological sections were taken. The changes of lung inflammation were observed by hematoxylin-eosin (HE) staining. EOS count and cytokines IL-4, IL-5, IL-10, IL-12, transforming growth factor beta (TGF-beta) were detected in BALF of mice, serum levels of OVA specific IgE antibodies were detected, and spleen cell culture supernatant (SPC) was collected for cytokines IL-4, IL-5, IL-10, IL-12, TGF-beta detection. HE staining results showed that compared with B, C, D, E group, the inflammation of lung tissue in F group was significantly reduced, and the total score of lung histopathology was statistically significant (P 0.05); the percentage of EOS counts in BALF of A, B, C, D, E, F group was (8.40 (+ 1.14)), (40.00 (+ 3.39)), (23.67 (+ 4.62)), (25.20 (+ 3.22)), (13.00 (+ 1.58)), respectively, compared with B, C, D, E group. Compared with F group, the number of EOSin F group decreased significantly (P 0.05), the levels of serum OVA-specific IgE in A, B, C, C, D, D, E, F groups were (204.90 (+3.58) pg/ml, (1127.83 (+18.86) pg/ml, (1127.83 ((1127.83 (+18.86) pg/ml, (1097.03 (+1097.03 [(10 97.03 (+25.03 [) 25.80) pg/ml, (1097.08 [(1097.08 (10 97.08 [10 09.08 [10 09.08 (1 1039.78 [2.78 [2.90) pg/ml, (1 Six small groups, C, D, E, F The secretion levels of IL-4 in BALF of BALF in rats were (10.17 (+ 4.49) pg/ml, (80.36 (+ 10.94) pg/ml, (80.36 (+ 10.94) pg/ml, (80.36 (+10.94) pg/ml, (67.83 (+ 6.81) pg/ml, (67.83 (62.83 (+6.83 (+6.81,, (62.41 +4.02) pg/ml, (62.41 +4.02) pg/ml, (57.33 (+ 11.33 +11.82) pg/ml, (57.33 +11.82) pg/ml, (22.36 (+ 7.36 g / ml, (811.16 + 11.61) pg / ml, (313.77 + 2.97) pg / ml; The secretion levels of IL-10 were (120.79 +21.48) pg/ml, (72.77 +17.60) pg/ml, (72.77 +17.60) pg/ml, (81.63 [81.63 [9.08] pg/ml, (82.80 [2394] pg/ml, (82.80 [(82.80 [2394) pg/ml, (106.94 [106.94 [26.31) pg/ml, (248.89 [33.20) pg/ml, (248.89 [33.20) pg/ml, (248.37 [(1.53) pg/ml, (46.70 [7.00] pg/ml, (122.70 53 + 4.81 pg / ml, (340.95 + 22.22) pg / ml; TGF The levels of IL-P secretion were (176.91 [(176.91 [51.25) pg/ml, (190.09 [(190.09 [20.64] pg/ml, (190.09 [20.64 [(109.64 [34.71] pg/ml, (162.27 [38.95) pg/ml, (220.09 [25.00] pg/ml, (220.09 [(220.09 [25.09]) pg/ml, (966.82 [57.82 [57.48] pg/ml, (966.82 [57.82] 48] pg/ml, respectively). Compared with B, C, IL-4 secretion level in SPC of mice in F 6 groups The secretion levels of IL-5 and IL-5 were (12.13 [(2.13 [2.96] pg / ml, (17.77 [(248.77 [18.85 [(248.77 [(248.77 [18.85] pg / ml, (220.51 [11.84 [18.85] pg / ml, (220.51 [11.51 [11.84] 84] pg / ml, (217.13 [(217.13.13 [50.37 [50.37] pg / ml, (172.17.17.17.13 [70 [27.70 [27.81] pg / ml, (172.70 [76.70 96 The average values were (122.88 [(122.88 +16.81) pg/ml, (131.10 [(131.10 [11.52) pg/ml, (161.21 [(161.21 [1.80) pg/ml, (204.91 [(8.71) pg/ml, (220.90 [26.86) pg/ml, (220.90 [(220.90 [26.86 [998.81 [81 [138.46] pg/ml, (99.81 [138.46] pg/ml, (99.03 [5.13] 5.13] pg/ml, (66.03 + 2.31, (66.03 + 2.31) pg/9.53 + 16.74 pg / MK (405.70 + 57.66) pg / ml; TGF - beta secretion The levels of IL-4, IL-5, IL-4, IL-5, IL-4, IL-5, IL-5, IL-10, IL-10, IL-10, IL-12, IL-12, IL-betwere significantly increased (P 0.05), IL-10, IL-10, IL-12, IL-12, IL-10, IL-12, F-betlevels were significantly increased (P 0.05). Conclusion Compared with B, C, C, D, D and E groups, the levels of TG-4, IL-4, IL-5, IL-5, IL-5, IL-10, IL-10, IL-12, IL-12, IL-12, F-betwere significantly increased (P 0.05).allergy Asthma is inhibited by CD8 alpha -DC subgroup.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R383.24;R562.25
[Abstract]:Objective To investigate the role and possible mechanism of DC subpopulation immunized with SEA in inhibiting allergic asthma by adoptive transfer of soluble egg antigen (SEA) from Schistosoma japonicum.
Methods BALB/c mice were injected with SEA 50 ug/mouse via abdominal cavity and footpad once a week for 4 times.The control group was injected with normal saline.The spleen CD11c+CD8a+DC and CD11c+CD8a-DC subgroups were separated by immunomagnetic beads coated with antibody.Another 30 BALB/c mice were randomly divided into 6 groups.Group A was normal control group(SCN),group B was simple asthma group(SNO),and group C was adoptive transfer. CD11c+CD8a+DC group (AA), CD11c+CD8a-DC group (AB), SEA-immunized CD11c+CD8a+DC group (SA), SEA-immunized CD11c+CD8a+DC group (SB). C. D group mice were subjected to adoptive transfer of CD11c+CD8a+DC and'5x105 saline control CD11c+C+CD8a+DC via caudal vein, respectively. D8a-DC; E, F mice were immunized with 5 *105 CD11c+CD8a+DC and 5 *105 SEA-immunized CD11c+CD8a-TC respectively by tail vein adoptive transfer SEA. At the same time, B, C, D, E, F mice were immunized with ovalbumin (OVA) to induce asthma. Four weeks later, the lung tissues were dissected and pathological sections were taken. The changes of lung inflammation were observed by hematoxylin-eosin (HE) staining. EOS count and cytokines IL-4, IL-5, IL-10, IL-12, transforming growth factor beta (TGF-beta) were detected in BALF of mice, serum levels of OVA specific IgE antibodies were detected, and spleen cell culture supernatant (SPC) was collected for cytokines IL-4, IL-5, IL-10, IL-12, TGF-beta detection. HE staining results showed that compared with B, C, D, E group, the inflammation of lung tissue in F group was significantly reduced, and the total score of lung histopathology was statistically significant (P 0.05); the percentage of EOS counts in BALF of A, B, C, D, E, F group was (8.40 (+ 1.14)), (40.00 (+ 3.39)), (23.67 (+ 4.62)), (25.20 (+ 3.22)), (13.00 (+ 1.58)), respectively, compared with B, C, D, E group. Compared with F group, the number of EOSin F group decreased significantly (P 0.05), the levels of serum OVA-specific IgE in A, B, C, C, D, D, E, F groups were (204.90 (+3.58) pg/ml, (1127.83 (+18.86) pg/ml, (1127.83 ((1127.83 (+18.86) pg/ml, (1097.03 (+1097.03 [(10 97.03 (+25.03 [) 25.80) pg/ml, (1097.08 [(1097.08 (10 97.08 [10 09.08 [10 09.08 (1 1039.78 [2.78 [2.90) pg/ml, (1 Six small groups, C, D, E, F The secretion levels of IL-4 in BALF of BALF in rats were (10.17 (+ 4.49) pg/ml, (80.36 (+ 10.94) pg/ml, (80.36 (+ 10.94) pg/ml, (80.36 (+10.94) pg/ml, (67.83 (+ 6.81) pg/ml, (67.83 (62.83 (+6.83 (+6.81,, (62.41 +4.02) pg/ml, (62.41 +4.02) pg/ml, (57.33 (+ 11.33 +11.82) pg/ml, (57.33 +11.82) pg/ml, (22.36 (+ 7.36 g / ml, (811.16 + 11.61) pg / ml, (313.77 + 2.97) pg / ml; The secretion levels of IL-10 were (120.79 +21.48) pg/ml, (72.77 +17.60) pg/ml, (72.77 +17.60) pg/ml, (81.63 [81.63 [9.08] pg/ml, (82.80 [2394] pg/ml, (82.80 [(82.80 [2394) pg/ml, (106.94 [106.94 [26.31) pg/ml, (248.89 [33.20) pg/ml, (248.89 [33.20) pg/ml, (248.37 [(1.53) pg/ml, (46.70 [7.00] pg/ml, (122.70 53 + 4.81 pg / ml, (340.95 + 22.22) pg / ml; TGF The levels of IL-P secretion were (176.91 [(176.91 [51.25) pg/ml, (190.09 [(190.09 [20.64] pg/ml, (190.09 [20.64 [(109.64 [34.71] pg/ml, (162.27 [38.95) pg/ml, (220.09 [25.00] pg/ml, (220.09 [(220.09 [25.09]) pg/ml, (966.82 [57.82 [57.48] pg/ml, (966.82 [57.82] 48] pg/ml, respectively). Compared with B, C, IL-4 secretion level in SPC of mice in F 6 groups The secretion levels of IL-5 and IL-5 were (12.13 [(2.13 [2.96] pg / ml, (17.77 [(248.77 [18.85 [(248.77 [(248.77 [18.85] pg / ml, (220.51 [11.84 [18.85] pg / ml, (220.51 [11.51 [11.84] 84] pg / ml, (217.13 [(217.13.13 [50.37 [50.37] pg / ml, (172.17.17.17.13 [70 [27.70 [27.81] pg / ml, (172.70 [76.70 96 The average values were (122.88 [(122.88 +16.81) pg/ml, (131.10 [(131.10 [11.52) pg/ml, (161.21 [(161.21 [1.80) pg/ml, (204.91 [(8.71) pg/ml, (220.90 [26.86) pg/ml, (220.90 [(220.90 [26.86 [998.81 [81 [138.46] pg/ml, (99.81 [138.46] pg/ml, (99.03 [5.13] 5.13] pg/ml, (66.03 + 2.31, (66.03 + 2.31) pg/9.53 + 16.74 pg / MK (405.70 + 57.66) pg / ml; TGF - beta secretion The levels of IL-4, IL-5, IL-4, IL-5, IL-4, IL-5, IL-5, IL-10, IL-10, IL-10, IL-12, IL-12, IL-betwere significantly increased (P 0.05), IL-10, IL-10, IL-12, IL-12, IL-10, IL-12, F-betlevels were significantly increased (P 0.05). Conclusion Compared with B, C, C, D, D and E groups, the levels of TG-4, IL-4, IL-5, IL-5, IL-5, IL-10, IL-10, IL-12, IL-12, IL-12, F-betwere significantly increased (P 0.05).allergy Asthma is inhibited by CD8 alpha -DC subgroup.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R383.24;R562.25
【参考文献】
相关期刊论文 前7条
1 熊涛;万灿;倪凤娥;张枫;;日本血吸虫未成熟虫卵的制备[J];长江大学学报(自然科学版);2011年04期
2 尹静波;黄永富;;炎性细胞因子在支气管哮喘发病机制中的作用[J];国际检验医学杂志;2011年10期
3 徐曼;徐光绪;王渝琦;蒋小卫;戴明q，
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