重组5型腺病毒处理不会加重卵清蛋白诱发哮喘小鼠的哮喘症状
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摘要
目的探索重组5型腺病毒(rAd5)对卵清蛋白(OVA)诱发的小鼠哮喘模型的影响。方法 60只6~8周龄、体质量18~20 g的雌性C57BL/6小鼠被随机分为健康对照组、rAd5对照组、OVA哮喘对照组、rAd5处理的OVA哮喘组,每组15只。OVA哮喘对照组、rAd5处理的OVA哮喘组在第0、7、14天分别采用含50μg OVA和2 mg氢氧化铝混悬液腹腔注射致敏3次、于第42、43、44天连续3 d用OVA雾化激发制作哮喘模型。其余两组采用相同体积的生理盐水处理。rAd5对照组、rAd5处理的OVA哮喘组于实验第21天开始肌注rAd5(2.5×109个病毒颗粒/只)1次。并于第35天滴鼻加强1次rAd5(2.5×109个病毒颗粒/只)。所有小鼠于最后一次雾化激发48 h内进行安乐死。通过无创肺功能仪体描法检测小鼠气道高反应,收集血清及肺泡灌洗液标本、收集肺组织用于HE及PAS染色,通过ELISA检测肺泡灌洗液中的白细胞介素4(IL-4)、IL-5、IL-13及血清中的总IgE。结果与健康对照组及rAd5对照组相比,OVA哮喘对照组、rAd5处理的OVA哮喘组其肺部炎症及黏液分泌显著增加,其肺泡灌洗液中IL-4、IL-5、IL-13及血清中的总IgE含量显著增高,其气道高反应显著升高。然而,OVA哮喘对照组与rAd5处理的OVA哮喘组之间无显著差异。结论重组5型腺病毒作用于小鼠哮喘模型不会加重哮喘症状。
Objective To explore the effect of recombinant adenovirus type 5( rAd5) on a mouse model of ovalbumin( OVA)-induced asthma. Methods Sixty female C57BL/6 mice( 6- 8 weeks old and 18- 20 g) were randomly divided into4 groups: healthy control group,rAd5 control group,OVA asthma control group and OVA asthma group with rAd5 immunization.15 mice in each group. The mice in the OVA asthma control group and the OVA asthma group with rAd5 immunization were sensitized by intraperitoneal injection of 50 μg OVA emulsified in 2 mg of aluminum potassium sulfate on day 0,7 and 14;subsequently,the mice in the two groups were exposed to OVA aerosol challenge for 3 consecutive days( on day 42,43 and44). Meanwhile,mice in the healthy control group and the rAd5 control group were given an equal volume of normal saline.What's more,mice in the OVA asthma group with rAd5 immunization and the rAd5 control group were immunized intramuscularly with 2. 5 × 109 viral particles( VP) of rAd5 vectors on day 21,and were boosted intranasally with the same dose of rAd5 vectors on day 35. Al mice were euthanized within 48 hours after the last chal enge. The airway hyper-responsiveness( AHR) of mice was measured by single-chamber,whole-body plethysmography. The blood was obtained,the samples of bronchoalveolar lavage fluid( BALF) were collected,and lung tissues were prepared for HE and PAS staining. The levels of interleukin-4( IL-4),IL-5,IL-13 in BALF and total IgE in serum were detected by ELISA. Results Compared with the healthy control group and the rAd5 control group,the pulmonary inflammation and mucus production significantly increased in the OVA asthma control group and the OVA asthma group with rAd5 immunization. The levels of IL-4,IL-5,IL-13 in BALF,total serum IgE and the AHR were also significantly enhanced in the OVA asthma control group and the OVA asthma group with rAd5 immunization. However,no significant difference was showed between the OVA asthma control group and the OVA asthma group with rAd5 immunization. Conclusion The use of recombinant adenovirus type 5 does not exacerbate asthma symptoms in a mouse model of OVA-induced asthma.
Objective To explore the effect of recombinant adenovirus type 5( rAd5) on a mouse model of ovalbumin( OVA)-induced asthma. Methods Sixty female C57BL/6 mice( 6- 8 weeks old and 18- 20 g) were randomly divided into4 groups: healthy control group,rAd5 control group,OVA asthma control group and OVA asthma group with rAd5 immunization.15 mice in each group. The mice in the OVA asthma control group and the OVA asthma group with rAd5 immunization were sensitized by intraperitoneal injection of 50 μg OVA emulsified in 2 mg of aluminum potassium sulfate on day 0,7 and 14;subsequently,the mice in the two groups were exposed to OVA aerosol challenge for 3 consecutive days( on day 42,43 and44). Meanwhile,mice in the healthy control group and the rAd5 control group were given an equal volume of normal saline.What's more,mice in the OVA asthma group with rAd5 immunization and the rAd5 control group were immunized intramuscularly with 2. 5 × 109 viral particles( VP) of rAd5 vectors on day 21,and were boosted intranasally with the same dose of rAd5 vectors on day 35. Al mice were euthanized within 48 hours after the last chal enge. The airway hyper-responsiveness( AHR) of mice was measured by single-chamber,whole-body plethysmography. The blood was obtained,the samples of bronchoalveolar lavage fluid( BALF) were collected,and lung tissues were prepared for HE and PAS staining. The levels of interleukin-4( IL-4),IL-5,IL-13 in BALF and total IgE in serum were detected by ELISA. Results Compared with the healthy control group and the rAd5 control group,the pulmonary inflammation and mucus production significantly increased in the OVA asthma control group and the OVA asthma group with rAd5 immunization. The levels of IL-4,IL-5,IL-13 in BALF,total serum IgE and the AHR were also significantly enhanced in the OVA asthma control group and the OVA asthma group with rAd5 immunization. However,no significant difference was showed between the OVA asthma control group and the OVA asthma group with rAd5 immunization. Conclusion The use of recombinant adenovirus type 5 does not exacerbate asthma symptoms in a mouse model of OVA-induced asthma.
引文
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[2]Martinez F D,Vercelli D.Asthma[J].Lancet,2013,382(9901):1360-1372.
[3]Carr T F,Beamer P I,Rothers J,et al.Prevalence of asthma in school children on the Arizona-Sonora border[J/OL].J Allergy Clin Immunol Pract,2016 Aug 17.pii:S2213-2198(16)30267-7.doi:10.1016/j.jaip.2016.07.001.[Epub ahead of print].
[4]Maselli D J,Velez M I,Rogers L.Reslizumab in the management of poorly controlled asthma:the data so far[J].J Asthma Allergy,2016,9:155-162.
[5]Noval Rivas M,Chatila T A.Regulatory T cells in allergic diseases[J].J Allergy Clin Immunol,2016,138(3):639-652.
[6]Kim E H,Park H J,Han G Y,et al.Intranasal adenovirus-vectored vaccine for induction of long-lasting humoral immunity-mediated broad protection against influenza in mice[J].J Virol,2014,88(17):9693-9703.
[7]Afkhami S,Yao Y,Xing Z.Methods and clinical development of adenovirus-vectored vaccines against mucosal pathogens[J/OL].Mol Ther Methods Clin Dev,2016,3:16030.doi:10.1038/mtm.2016.30.eC ollection 2016.Review
[8]De Santis O,Audran R,Pothin E,et al.Safety and immunogenicity of a chimpanzee adenovirus-vectored Ebola vaccine in healthy adults:a randomised,double-blind,placebo-controlled,dose-finding,phase1/2a study[J].Lancet Infect Dis,2016,16(3):311-320.
[9]Hauptmann M,Schaible U E.Linking microbiota and respiratory disease[J/OL].FEBS Lett,2016,590(21):3721-3738.
[10]Smet M,Van Hoecke L,De Beuckelaer A,et al.Cholesterol-sensing liver X receptors stimulate Th2-driven allergic eosinophilic asthma in mice[J].Immun Inflamm Dis,2016,4(3):350-361.
[11]Abel B,Tameris M,Mansoor N,et al.The novel tuberculosis vaccine,AERAS-402,induces robust and polyfunctional CD4+and CD8+T cells in adults[J].Am J Respir Crit Care Med,2010,181(12):1407-1417.
[12]Zhang Y,Feng L,Li L,et al.Effects of the fusion design and immunization route on the immunogenicity of Ag85A-Mtb32 in adenoviral vectored tuberculosis vaccine[J].Hum Vaccin Immunother,2015,11(7):1803-1813.
[13]Ge X N,Ha S G,Greenberg Y G,et al.Regulation of eosinophilia and allergic airway inflammation by the glycan-binding protein galectin-1[J].Proc Natl Acad Sci U S A,2016,113(33):E4837-E4846.
[14]Lafkas D,Shelton A,Chiu C,et al.Therapeutic antibodies reveal Notch control of transdifferentiation in the adult lung[J].Nature,2015,528(7580):127-131.
[15]Garbani M,Xia W,Rhyner C,et al.Allergen-loaded strontium-doped hydroxyapatite spheres improve allergen-specific immunotherapy in mice[J/OL].Allergy,2016 Sep 3.doi:10.1111/all.13041.[Epub ahead of print].
[16]Ren Y,Li C,Feng L,et al.Proton channel activity of influenza a virus matrix protein 2 contributes to autophagy arrest[J].J Virol,2015,90(1):591-598.
[17]Sun C,Chen Z,Tang X,et al.Mucosal priming with a replicatingvaccinia virus-based vaccine elicits protective immunity to simian immunodeficiency virus challenge in rhesus monkeys[J].J Virol,2013,87(10):5669-5677.
[18]Sun C,Feng L,Zhang Y,et al.Circumventing antivector immunity by using adenovirus-infected blood cells for repeated application of adenovirus-vectored vaccines:proof of concept in rhesus macaques[J].J Virol,2012,86(20):11031-11042.
[19]Martín V,Pascual E,Avia M,et al.Protective efficacy in sheep of adenovirus-vectored vaccines against bluetongue virus is associated with specific T cell responses[J/OL].PLoS One,2015,10(11):e0143273.doi:10.1371/journal.pone.0143273.eC ollection2015.
[20]Wong G,Richardson J S,Pillet S,et al.Adenovirus-vectored vaccine provides postexposure protection to Ebola virus-infected nonhuman primates[J/OL].J Infect Dis,2015,212 Suppl 2:S379-S383.doi:10.1093/infdis/jiv102.Epub 2015 May 9.
[21]Xiang K,Ying G,Yan Z,et al.Progress on adenovirus-vectored universal influenza vaccines[J].Hum Vaccin Immunother,2015,11(5):1209-1222.
[22]Jeyanathan M,Thanthrige-Don N,Afkhami S,et al.Novel chimpanzee adenovirus-vectored respiratory mucosal tuberculosis vaccine:overcoming local anti-human adenovirus immunity for potent TB protection[J].Mucosal Immunol,2015,8(6):1373-1387.
[23]Ruan G,Tao B,Wang D,et al.Chinese herbal medicine formula Gu-Ben-Fang-Xiao-Tang attenuates airway inflammation by modulating Th17/Treg balance in an ovalbumin-induced murine asthma model[J].Exp Ther Med,2016,12(3):1428-1434.
[24]Wei D Z,Guo X Y,Lin L N,et al.Effects of angelicin on ovalbumin(ova)-induced airway inflammation in a mouse model of asthma[J].Inflammation,2016,39(6):1876-1882.
[25]Cui W,Zhang P,Gu J,et al.Vitamin a deficiency promotes inflammation by induction of type 2 cytokines in experimental ovalbumin-induced asthma murine model[J].Inflammation,2016,39(5):1798-1804.
[26]Rana S,Shahzad M,Shabbir A,et al.Pistacia integerrima ameliorates airway inflammation by attenuation of TNF-α,IL-4,and IL-5 expression levels,and pulmonary edema by elevation of AQP1 and AQP5expression levels inmouse model of ovalbumin-induced allergic asthma[J].Phytomedicine,2016,23(8):838-845.
[27]Wei Y,Liu J,Zhang H,et al.Ligustrazine attenuates inflammation and the associated chemokines and receptors in ovalbumine-induced mouse asthma model[J].Environ Toxicol Pharmacol,2016,46:55-61.
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