中华耳鼻咽喉头颈外科杂志
中華耳鼻嚥喉頭頸外科雜誌
중화이비인후두경외과잡지
CHINESE JOURNAL OF OTORHINOLARYNGOLOGY HEAD AND NECK SURGERY
2013年
7期
563-567
,共5页
陈建军%王景慧%袁阳%付俊梅%余滋中%王彦君%孔维佳
陳建軍%王景慧%袁暘%付俊梅%餘滋中%王彥君%孔維佳
진건군%왕경혜%원양%부준매%여자중%왕언군%공유가
卵白蛋白%变应原%超敏反应%T淋巴细胞,辅助诱导%免疫记忆%细胞因子类%小鼠%动物,新生
卵白蛋白%變應原%超敏反應%T淋巴細胞,輔助誘導%免疫記憶%細胞因子類%小鼠%動物,新生
란백단백%변응원%초민반응%T림파세포,보조유도%면역기억%세포인자류%소서%동물,신생
Ovalbumin%Allergens%Hypersensitivity%T-lymphocytes,helper-inducer%Immunologic memory%Cytokines%Mice%Animals,newborn
目的 研究小鼠生命早期不同剂量变应原免疫干预对将来免疫功能状态的影响.方法 50只新生小鼠分完全随机为4组,出生后第1、8、15天给予不同剂量卵清蛋白(ovalbumin,OVA)或生理盐水及佐剂皮下注射.NS组:10只,皮下注射生理盐水；NS+ AL组:10只,皮下注射生理盐水+ Al(OH)3；小剂量组:15只,皮下注射10 μg OVA +AL(OH)3;大剂量组:15只,皮下注射1000 μg OVA +Al(OH)3.5周龄时尾静脉注射激发,其中NS组及NS+ AL组给予生理盐水激发,小剂量组及大剂量组予100 μg OVA激发,1周后收集血清,检测OVA特异性IgE、IgG1及IgG2a含量.行脾单个核细胞培养后检测上清中白细胞介素(interleukin,IL)4、γ干扰素(interferon,IFN-γ)、IL-10水平.流式细胞术检测脾单个核细胞中CD4+ IL4+、CD4+ IFN-γ+、CD4+ IL-10+细胞表达情况.结果 小剂量组血清OVA特异性IgE为(0.33 ±0.18),高于NS组的(0.07 ±0.01)、NS+ AL组的(0.09 ±0.04)、以及大剂量组的(0.17±0.10),差异有统计学意义(t值分别为-3.46、-3.21、2.58,P值分别＜0.01,＜0.01,＜0.05).大剂量组OVA-IgE高于NS组(t=-2.53,P＜0.05)但与NS+ AL组间差异无统计学意义(t=-2.04,P＞0.05).小剂量组及大剂量组OVA特异性IgG1及IgG2a均高于NS及NS+ AL组(P值均＜0.05).脾细胞培养细胞因子检测结果显示,小剂量组IL-4、IFN-γ及IL-10均高于NS及NS+ AL组(P值均＜0.01).但小剂量组IFN-γ/IL-4比值低于NS及NS+ AL组(t值分别为2.14、3.44,P值均＜0.05),而大剂量组IFN-γ/IL-4比值高于NS及NS+ AL组(t值分别为-2.14、-1.61,P值均＜0.05).流式检测结果表明,大剂量组CD4+ IL-4+细胞较小剂量组明显减少(P＜0.05),与NS组及NS+ AL组差异无统计学意义(P＞0.05).结论 生命早期小剂量OVA干预,可使小鼠机体形成Th2方向为主导的特异性免疫反应；而大剂量OVA干预时,则可使其形成Th1方向为主导的特异性免疫反应.
目的 研究小鼠生命早期不同劑量變應原免疫榦預對將來免疫功能狀態的影響.方法 50隻新生小鼠分完全隨機為4組,齣生後第1、8、15天給予不同劑量卵清蛋白(ovalbumin,OVA)或生理鹽水及佐劑皮下註射.NS組:10隻,皮下註射生理鹽水；NS+ AL組:10隻,皮下註射生理鹽水+ Al(OH)3；小劑量組:15隻,皮下註射10 μg OVA +AL(OH)3;大劑量組:15隻,皮下註射1000 μg OVA +Al(OH)3.5週齡時尾靜脈註射激髮,其中NS組及NS+ AL組給予生理鹽水激髮,小劑量組及大劑量組予100 μg OVA激髮,1週後收集血清,檢測OVA特異性IgE、IgG1及IgG2a含量.行脾單箇覈細胞培養後檢測上清中白細胞介素(interleukin,IL)4、γ榦擾素(interferon,IFN-γ)、IL-10水平.流式細胞術檢測脾單箇覈細胞中CD4+ IL4+、CD4+ IFN-γ+、CD4+ IL-10+細胞錶達情況.結果 小劑量組血清OVA特異性IgE為(0.33 ±0.18),高于NS組的(0.07 ±0.01)、NS+ AL組的(0.09 ±0.04)、以及大劑量組的(0.17±0.10),差異有統計學意義(t值分彆為-3.46、-3.21、2.58,P值分彆＜0.01,＜0.01,＜0.05).大劑量組OVA-IgE高于NS組(t=-2.53,P＜0.05)但與NS+ AL組間差異無統計學意義(t=-2.04,P＞0.05).小劑量組及大劑量組OVA特異性IgG1及IgG2a均高于NS及NS+ AL組(P值均＜0.05).脾細胞培養細胞因子檢測結果顯示,小劑量組IL-4、IFN-γ及IL-10均高于NS及NS+ AL組(P值均＜0.01).但小劑量組IFN-γ/IL-4比值低于NS及NS+ AL組(t值分彆為2.14、3.44,P值均＜0.05),而大劑量組IFN-γ/IL-4比值高于NS及NS+ AL組(t值分彆為-2.14、-1.61,P值均＜0.05).流式檢測結果錶明,大劑量組CD4+ IL-4+細胞較小劑量組明顯減少(P＜0.05),與NS組及NS+ AL組差異無統計學意義(P＞0.05).結論 生命早期小劑量OVA榦預,可使小鼠機體形成Th2方嚮為主導的特異性免疫反應；而大劑量OVA榦預時,則可使其形成Th1方嚮為主導的特異性免疫反應.
목적 연구소서생명조기불동제량변응원면역간예대장래면역공능상태적영향.방법 50지신생소서분완전수궤위4조,출생후제1、8、15천급여불동제량란청단백(ovalbumin,OVA)혹생리염수급좌제피하주사.NS조:10지,피하주사생리염수；NS+ AL조:10지,피하주사생리염수+ Al(OH)3；소제량조:15지,피하주사10 μg OVA +AL(OH)3;대제량조:15지,피하주사1000 μg OVA +Al(OH)3.5주령시미정맥주사격발,기중NS조급NS+ AL조급여생리염수격발,소제량조급대제량조여100 μg OVA격발,1주후수집혈청,검측OVA특이성IgE、IgG1급IgG2a함량.행비단개핵세포배양후검측상청중백세포개소(interleukin,IL)4、γ간우소(interferon,IFN-γ)、IL-10수평.류식세포술검측비단개핵세포중CD4+ IL4+、CD4+ IFN-γ+、CD4+ IL-10+세포표체정황.결과 소제량조혈청OVA특이성IgE위(0.33 ±0.18),고우NS조적(0.07 ±0.01)、NS+ AL조적(0.09 ±0.04)、이급대제량조적(0.17±0.10),차이유통계학의의(t치분별위-3.46、-3.21、2.58,P치분별＜0.01,＜0.01,＜0.05).대제량조OVA-IgE고우NS조(t=-2.53,P＜0.05)단여NS+ AL조간차이무통계학의의(t=-2.04,P＞0.05).소제량조급대제량조OVA특이성IgG1급IgG2a균고우NS급NS+ AL조(P치균＜0.05).비세포배양세포인자검측결과현시,소제량조IL-4、IFN-γ급IL-10균고우NS급NS+ AL조(P치균＜0.01).단소제량조IFN-γ/IL-4비치저우NS급NS+ AL조(t치분별위2.14、3.44,P치균＜0.05),이대제량조IFN-γ/IL-4비치고우NS급NS+ AL조(t치분별위-2.14、-1.61,P치균＜0.05).류식검측결과표명,대제량조CD4+ IL-4+세포교소제량조명현감소(P＜0.05),여NS조급NS+ AL조차이무통계학의의(P＞0.05).결론 생명조기소제량OVA간예,가사소서궤체형성Th2방향위주도적특이성면역반응；이대제량OVA간예시,칙가사기형성Th1방향위주도적특이성면역반응.
Objective To study the effect of neonatal immunization with different dosage allergen on the immunity of mice when grown.Methods Fifty neonatal BALB/c mice were divided into 4 groups randomly and subcutaneous injected with different dosage of ovalbumin (OVA) on day 1,8 and 15 after born [NS group(10):injected with saline alone ; NS + AL group(10):injected with saline and AL(OH) 3 ; small dosage (SD) group (15):injected with 10 μg OVA and AL (OH) 3 ; large dosage (LD) group (15):injected with 1000 μg OVA and AL(OH)3].The mice were then challenged using caudal vein injection on 5 weeks old (NS group and NS + AL group were challenged with saline,SD group and LD group were challenged with 100 μg OVA).The blood was collected 1 week later to examine OVA specific IgE,IgG1 and IgG2a.Mononuclear cells were drawn from the spleen and cultured.Concentration of IL-4,IFN-r,IL-10 was examined in the cultural supernatant.Flow cytometry was used to test the expression of CD4 + IL-4 +,CD4 + IFN-γ+,CD4 + IL-10 + cells.Results It was found that concentration of OVA specific IgE (OVA-sIgE) in SD group (0.33 ± 0.18) was significantly higher than that of NS (0.07 ± 0.01) and NS + AL (0.09±0.04) group (t value was-3.46 and-3.21,all P＜0.01),and LD group (0.17 ±0.10) as well (t =2.58,P ＜0.05).The concentration of OVA-sIgE was higher in LD group than that of NS group (t=-2.53,P＜0.05),but similar with that of NS +AL group (t =-2.04,P ＞0.05).Both the concentration of OVA-sIgG1 and sIgG2a was higher in SD and LD group than that of NS and NS + AL group (all P ＜ 0.05).The concentration of IL-4,IFN-γ and IL-10 in the cultural supernatant of spleen mononuclear was all higher in SD group than that of NS and NS + AL group (all P ＜ 0.01).The ratio of IFN-γ/IL-4 was significantly lower in SD group than that of NS and NS + AL group (t value was 2.14,3.44,all P ＜ 0.05),while the same ratio was higher in LD group than that of NS and NS + AL group (t value was-2.14,-1.61,all P ＜0.05).Ratio of CD4+ IL-4+ cells was significant lower in LD group than that of SD group (P ＜ 0.05),while it was not different with that of NS and NS + AL group (P ＞ 0.05).Conclusion Neonatal immunization with low dosage OVA could generate a specific immunity with Th2 direction,while with large dosage OVA could generate a specific immunity with Th1 direction.
