【摘要】：目的通過過繼轉(zhuǎn)移日本血吸蟲可溶性蟲卵抗原(SEA)免疫小鼠樹突狀細(xì)胞(DC)亞群,探討SEA免疫的DC亞群在抑制過敏性哮喘中的作用及可能的作用機制。 方法BALB/c小鼠經(jīng)腹腔及足墊注射SEA50μg/只,每周1次,共4次。對照組注射生理鹽水。用抗體包被的免疫磁珠分離小鼠脾CD11c+CD8α+DC與CD11c+CD8α-DC亞群。另取30只BALB/c小鼠,隨機分為6組,A組為正常對照組(SCN),B組為單純哮喘組(SNO),C組為過繼轉(zhuǎn)移鹽水對照CD11c+CD8α+DC組(AA), D組為過繼轉(zhuǎn)移鹽水對照CD11c+CD8α-DC組(AB),E組為過繼轉(zhuǎn)移SEA免疫CD11c+CD8α+DC組(SA),F組為過繼轉(zhuǎn)移SEA免疫CD11c+CD8α-DC組(SB)。C、D組小鼠分別經(jīng)尾靜脈過繼轉(zhuǎn)移5×105個鹽水對照CD11c+CD8α+DC和'5x105個鹽水對照CD11c+CD8α-DC; E、F組小鼠分別經(jīng)尾靜脈過繼轉(zhuǎn)移SEA免疫5×105個CD11c+CD8α+DC和5×105個SEA免疫CD11c+CD8α-TC。1h后B、C、D、E、F組小鼠同時用卵白蛋白(OVA)誘發(fā)哮喘,4周后剖殺,取肺組織,做病理切片,經(jīng)蘇木精-伊紅(HE)染色,光鏡下觀察肺組織炎癥變化；收集小鼠支氣管肺泡灌洗液(BALF)進行嗜酸性粒細(xì)胞(EOS)計數(shù)及細(xì)胞因子IL-4、IL-5、IL-10、IL-12、轉(zhuǎn)化生長因子β(TGF-β)檢測；收集血清檢測OVA特異性IgE抗體水平；收集脾細(xì)胞培養(yǎng)上清(SPC)進行細(xì)胞因子IL-4、IL-5、IL-10、IL-12、TGF-β檢測。結(jié)果肺組織病理切片HE染色結(jié)果顯示,與B、C、D、E組相比,F組小鼠肺組織炎癥明顯減輕,肺組織病理學(xué)總評分有統(tǒng)計學(xué)意義(P0.05)；A、B、C、D、E、F六組小鼠BALF中EOS計數(shù)百分比分別為(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),與B、C、D、E組比較,F組EOS數(shù)目明顯減少(P0.05)；A、B、C、D、E、F六組小鼠血清OVA特異性IgE水平分別為(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,與B、C、D、E組比較,F組IgE水平顯著下降(P0.05); A、B、C、D、E、F六組小鼠BALF中IL-4分泌水平分別為(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分泌水平分別為(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分泌水平分別為(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分泌水平分別為(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分泌水平分別為(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、與B、C、D、E組比較,F組IL-4、IL-5水平明顯降低(P0.05), IL-10、IL-12、TGF-β水平顯著升高(P0.05)。A、B、C、D、E、F六組小鼠SPC中IL-4分泌水平分別為(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分泌水平分別為(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分泌水平分別為(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分泌水平分別為(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分泌水平分別為(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;與B、C、D、E組比較,F組IL-4、IL-5水平明顯降低(P0.05), IL-10、IL-12、TGF-β水平顯著升高(P0.05)。結(jié)論日本血吸蟲SEA免疫的DC亞群對過敏性哮喘有抑制作用,該抑制作用主要通過CD8α-DC亞群產(chǎn)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.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R383.24;R562.25

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