转4-1BBL基因细胞诱导带瘤宿主抗瘤效应的体内研究

目的 观察协同刺激分子4-1BBLigand(4-1BBL)基因导入小鼠肝癌细胞Hepal-6后在小鼠体内诱导的抗瘤效应。方法 应用逆转录病毒载体，将小鼠4-1BBL导入小鼠肝癌细胞Hepal-6细胞中，Histidinol(HisD)筛选后获高表达4-1BBL分子阳性细胞克隆，经丝裂霉素C（MMC）处理制备成肿瘤细胞瘤苗TCV4-1BBL，观察其对不同动物模型的体内免疫保护作用和免疫治疗作用，结果 （1）能对同系肝癌细胞产生完全的免疫保护作用，并能保持无瘤状态长期存活（100d以上）；（2）对早期（接种7d)形成的肿瘤有强的治疗作用。（3）对晚期（接种14d)肿瘤，有明显的治疗作用。使大部分小鼠的肿瘤消退。结论 4-1BBL修饰的肿瘤细胞疫苗能明显刺激增强带瘤宿主的抗瘤效应。     

p33ING1转染HepG2肝癌细胞对p21WAF1/ CIP1表达的影响

目的 探讨p33ING1对pcDNA3.1-p33ING1转染后的HepG2肝癌细胞p21WAF1/CIP1表达的影响.方法 实验分为对照组和转染组;将pcDNA3.1-p33ING1及空载质粒pcDNA3.1转染到HepG2细胞中,最终建立稳定转染的细胞株,RT-PCR检测p33ING1表达;RT-PCR和免疫组化检...     

聚乙烯吡咯烷酮介导的白细胞介素-12基因治疗小鼠肝癌微小病灶的实验研究

目的 探讨白细胞介素(IL)-12基因注射对小鼠肝癌微小病灶的治疗作用。方法IL-12质粒转染BHK-21细胞，ELISA检测上清中IL-12的分泌，并用此上清刺激脾细胞，MTT检测其增殖。18只Balb／c小鼠股部肌肉内接种H22细胞后分为3组分别注射聚乙烯吡咯烷酮(PVP)／生理盐水(NS)、PVP／NS作溶剂的pcDNA3．1质粒和IL-2质粒观察肿瘤的形成；ELISA检测血清中IL-12的含量，切片观察肿瘤浸润淋巴细胞，并用免疫组化和TUNEL显示肿瘤血管密度和肿瘤细胞的凋亡。结果 IL-12因子可以分泌到胞外，促进脾淋巴细胞的增殖(15．2％)。与对照组相比，治疗组血清IL-12水平上升，瘤内淋巴细胞大量浸润，血管减少，肿瘤细胞凋亡，肝癌微小病灶生长受抑，抑制率高达85．7％。结论 PVP介导的IL-12基因重复注射对小鼠肝癌微小病灶具有很好的治疗效果。     

超声微泡包裹单纯疱疹病毒Ⅰ型胸苷激酶自杀基因靶向治疗小鼠肝癌的效果

目的 观察超声微泡携单纯疱疹病毒Ⅰ型胸苷激酶(HSV1-TK)基因在小鼠肝癌细胞H22移植瘤组织中的转染效果及联合更昔洛韦(GCV)后对肿瘤的抑瘤效应.方法 建立小鼠肝癌(H22)皮下移植瘤模型40只,随机分成4组,分别经鼠尾静脉注射入等量磷酸盐缓冲液(A组),HSV1-TK(B组)、HSV1-TK(C组)、HSV1-TK+微泡(D组),每次注射200μl,每隔3 d注射1次,共注射3次;对C组、D组小鼠分别给予1 MHz、2 W/cm~2、5 min超声辐照.首次注射HSV1TK 48 h后采用Western blot检测肿瘤组织中TK蛋白的表达情况,且各治疗组分别从腹腔注射GCV 0.2 ml(100 mg·kg~(-1)·d~(-1)),连续注射14 d.测肿瘤大小,计算抑瘤率,治疗结束后处死动物行病理学检查.数据采用重复测量资料的方差分析,两两比较采用LSD-t法.结果 在超声引导下微泡包裹的HSV1-TK基因可以准确定位于肿瘤组织并靶向释放,在处理组中均有TK蛋白的表达,其中D组表达量明显高于其他各组(P＜0.05).抑瘤率在A组、B组、C组、D组中分别为0、3.90%±1.80%、22.70%±2.86%、41.25%±3.20%(C组、D组与B组相比,P值均＜0.05;D组与C组相比,P＜0.05).HE染色结果显示D组较A组的坏死病变程度明显减轻.结论 超声辐照可破坏包裹HSV1-TK基冈的微泡以定位释放,既增加了肝癌基因治疗的靶向性,又提高了外源基因的转染效率,从而增强了自杀基因对小鼠肝癌的杀伤效果.     

miR-194-5p靶向MEX3A调控肝癌细胞活性和凋亡的机制

目的研究miR-194-5p对肝癌细胞活性和凋亡的影响,并探讨其机制。方法运用qRT-PCR法检测肝癌细胞HepG2、正常肝细胞L02中miR-194-5p和MEX3A的表达;根据分组:miR-194-5p组(转染miR-194-5p mimics)、miR-NC组(转染miR-NC)、si-MEX3A组(转染si-MEX3A)、si-con组(转染si-con)、miR-194-5p+pcDNA-MEX3A组(共转染miR-194-5p mimics和pcDNA-MEX3A)、miR-194-5p+pcDNA组(共转染miR-194-5p mimics和pcDNA),均用脂质体法将相关质粒转染至HepG2;噻唑蓝(MTT)法检测各组细胞的活性;流式细胞术检测各组细胞的凋亡;双荧光素酶报告基因检测实验检测各组细胞的荧光活性。结果与正常肝细胞L02相比,肝癌细胞HepG2中miR-194-5p表达明显降低,MEX3A表达明显升高(P0.05);过表达miR-194-5p、敲减MEX3A均可抑制HepG2细胞的活性,促进细胞凋亡;miR-194-5p可抑制野生型MEX3A细胞的荧光活性;过表达MEX3A能逆转miR-194-5p对HepG2细胞活性的抑制和凋亡的促进作用。结论 miR-194-5p可抑制肝癌细胞的活性,促进凋亡,其机制可能与靶向MEX3A有关,将可为肝癌的治疗提供新靶点。     

细小病毒H-1非结构蛋白NS1基因对人胃癌细胞的抑制作用研究

前期研究表明细小病毒H-1（H-1PV）对胃癌细胞生长具有一定抑制作用,非结构蛋白NSl可能是其细胞毒作用的效应蛋白。胃癌细胞CD44^＋群体中可能含有肿瘤干细胞。目的：探讨H-1PV非结构蛋白NSl基因对不同分化程度的人胃癌细胞的影响。方法：以双酶切和质粒测序鉴定真核表达质粒pcDNA3．1-NSl,采用脂质体法将重组质粒转染高分化胃癌MKN28细胞、中分化胃癌SGC7901细胞和低分化胃癌MKN45细胞,并设置空质粒转染对照。G418筛选稳定转染的细胞克隆．以RT-PCR法检测NSl基因表达、裸鼠体内成瘤实验检测NSl基因对不同分化程度胃癌细胞的作用．流式细胞术分析CD44表达。结果：重组质粒pcDNA3．1-NSl转染的三种胃癌细胞均稳定表达NSl基因。以10^6／300μl浓度的肿瘤细胞皮下接种裸鼠3周后,MKN28细胞空质粒转染组和重组质粒pcDNA3．1-NSl转染组均观察到肿瘤生长;SGC7901、MKN45细胞空质粒转染组形成瘤体,而重组质粒pcDNA3．1-NSl转染组未见瘤体。转染重组质粒pcDNA3．1-NSl后,MKN28细胞不表达CD44,SGC7901和MKN45细胞CD44表达明显降低。结论：H-1PV非结构蛋白NSl基因表达对中低分化的胃癌细胞有较好的抗肿瘤活性．可能与其降低胃癌干细胞表型CD44的表达有关。     

热碘油联合90Y瘤内注射治疗肝癌的实验研究

目的 探讨热碘油联合^90钇（^90Y）瘤内注射治疗肝癌的安全性和可行性。方法 将40只荷瘤昆明小鼠随机分为A、B、C、D组各10只。A组瘤内注射0．1ml热碘油与0．1ml（100μCi）^90Y混悬液,B组注射0．1ml热碘油;C组注射0．1ml（100μCi）^90Y;D组注射0．1ml生理盐水。A、C组注药后2h、3d、7d、10d、14d在γ相机和ECT扫描仪下观察放射性分布情况。各组在上述时间点抽血查肝、肾功能及血常规,并测肿瘤最大径a和最小径b,计算肿瘤体积及肿瘤生长率．同时取肿瘤组织作病理检查。结果 除肿瘤外,A、C组小鼠其他脏器内均未发现^90Y沉积．病理切片也未显示小鼠其他脏器因^90Y放射损伤所致的病理改变。A、B、C、D组小鼠肝、肾功能及血常规均无变化．未见有心、肺、脾、肾的病理改变及骨储抑制等。注药后第3天起,D组肿瘤生长迅速,生长率在3、7、10、14d时分别为13％、20％、29％、36％．与A组（分别为-11％、-19％、-29％、-38％）、B组（分别为-6％、-9％、-11％、-16％）、C组（分别为-1％、-3％、-6％、-9％）比较．差异均有显著性,P均〈0．01）各时间点A组生长率最低,与其他各组相比,差异有显著性,P均〈0．05）B组也低于C组,差异有显著性,P均〈0．05。D组在3～14d肿瘤坏死范围均未超过30％,A组肿瘤重度坏死率在3、7、10及14d时分别为58％、68％、77％及87％,与B、C、D组比较,差异有显著性,P均〈0．01;B、C、D组坏死率呈降低趋势,且组间差异有显著性,P均〈0．05。结论 热碘油联合^90Y瘤内注射治疗小鼠肝癌安全、有效。     

甲醛固定的H22细胞联合IL-2抑制小鼠H22肝癌生长效果观察

目的 观察甲醛固定的H22肿瘤细胞联合IL-2对小鼠H22肝癌细胞腹部移植瘤生长的影响.方法 20只小鼠,均采用股部皮下1×105个H22细胞的方法建立小鼠H22肝癌皮下移植瘤模型.将其随机分为A、B、C、D组,各5只.肿瘤直径超过1 cm后A、B、C、D组分别瘤内注射生理盐水、甲醛固定H22细胞、IL-2、甲醛固定H22细胞+IL-2各0.5 mL.H22细胞浓度为106/mL,IL-2浓度为8×103 IU/mL.每3d注射1次,共注射4次.治疗结束处死动物,取瘤称重,并计算抑瘤率.采用免疫组化SABC法对肿瘤组织染色,光景下观察瘤内CD8+T淋巴细胞表达情况.结果 A、B、C、D组肿瘤治疗后质量分别为(6.683 ±1.102)g、(2.851 ±0.223)g、(2.282 ±0.443)g、(1.052±0.225)g,B、C、D组与A组相比明显降低,D组与B、C组相比明显降低(P均＜0.05).B、C、D组抑瘤率分别为51.58％、60.54％、80.96％.D组小鼠瘤体内大量肿瘤细胞变性和坏死,且瘤内有大量CDs+T淋巴细胞浸润,阳性产物主要呈现在细胞膜上并被染成棕褐色.B、C组内仅见少量CD8+T淋巴细胞.结论 甲醛固定的H22细胞联合IL-2具有延迟荷瘤小鼠H22肝癌形成,限制生长的作用.     

微波消融联合5-氟尿嘧啶瘤内注射对小鼠结肠癌移植瘤的治疗

目的:研究微波消融(MA)联合5-氟尿嘧啶瘤内注射对小鼠移植性结肠癌的治疗作用.方法:Balb/c小鼠皮下接种结肠癌CT26细胞建立肿瘤模型,肿瘤分别给予瘤周注射PBS、MA、瘤周注射5-氟尿嘧啶和MA+瘤周注射5-氟尿嘧啶4种处理.测量各组肿瘤大小;观察肿瘤复发情况、动物生存期及生活状态;ELISA法检测各组小鼠外周...     

共刺激分子4-1BBL在过敏性支气管哮喘患者外周血的表达及其意义

目的 4-1BBL(CD137L)为肿瘤坏死因子超家族成员,4-1BBL与其受体4-1BB结合为T细胞活化提供共刺激信号.有研究证实激动性4-1BBL抗体能降低过敏性哮喘小鼠的气道高反应性,减少气道炎症细胞浸润.但4-1BBL在过敏性哮喘患者中的作用尚不清楚.方法 我们比较了过敏性哮喘患者和健康对照组外周血单个核细胞中...     

Correlation of 4-1BBL+ B Cells in Tumor Draining Lymph Nodes with Pathological Characteristics of Breast Cancer

Background: B cells can increase the expression of granzyme B in CD8+ T cells through 4-1BBL/4-1BB interaction and promote anti-tumor immunity. Objective: To investigate the expression of 4-1BBL on B cells in the breast tumor draining lymph nodes (TDLNs) and its association with disease parameters. Methods: Using Ficoll-Hypaque gradient centrifugation, mononuclear cells were isolated from axillary lymph nodes of 42 patients. Cells received 4 hours of PMA/Ionomycin stimulation, in vitro. Both unstimulated and stimulated cells were stained with anti‒CD19 and anti‒4-1BBL antibodies and subjected to flow cytometry. Results: 4-1BBL expression was detected on 2.8 ± 1.7% of unstimulated B cells, while 27.4 ± 11.9% of B cells expressed this co-stimulatory molecule following stimulation. In steady state, the percentage of 4-1BBL+ B cells was not associated with cancer characteristics. However, in patients with invasive ductal carcinoma, the percentage of 4-1BBL expressing B cells in stimulated condition had a decreasing trend in grade III, compared to grade II+I. In addition, significantly higher frequency of 4-1BBL+ B cells was seen in the TDLNs of ER+ or PR+ compared with ER‒ or PR‒ patients (p=0.021 and p=0.015, respectively). No significant associations were observed between the frequency of 4-1BBL+ B cells and the number of involved LNs, Her2 expression or disease stage. Conclusions: The frequency of 4-1BBL+ B cells significantly increased following a short time activation, and showed relative and significant associations with tumor grade and estrogen receptor status, respectively. More investigations are required to evaluate the potential of 4-1BBL+ B cells for use in immunotherapy.     

4-1BBL重组腺病毒载体的构建及表达

目的构建4-1BBL基因重组腺病毒载体,为前列腺癌的免疫治疗奠定基础。方法以质粒pcDNA3-m4-1BBL为模板,PCR法扩增出4-1BBL cDNA,并将其插入腺病毒穿梭质粒pAdTrack-CMV中构建成腺病毒穿梭质粒pAdTrackCMV-m4-1BBL,经PmeⅠ酶切线性化,与腺病毒骨架质粒pAdEasy-1共转化大肠杆菌BJ5183,挑选同源重组质粒,PacⅠ酶切线性化后,转染HEK293细胞包装成重组病毒颗粒,荧光显微镜观察绿色荧光表达,RT-PCR和Western blot法检测4-1BBL表达。结果目的基因经酶切分析,测序鉴定正确,RT-PCR和Western blot证实4-1BBL表达。结论成功构建了含4-1BBL基因的腺病毒载体,该载体可用于前列腺癌的免疫治疗。     

巨噬细胞炎性蛋白-1α联合4-1BB配体降低肝癌细胞体内致瘤性研究

目的 观察巨噬细胞炎性蛋白-1α(MIP-1α)联合4-1BB配体(4-1BB L)对肝癌细胞体内致瘤性的影响.方法 以小鼠4-1BB L(m4-1BB L)重组逆转录病毒感染Hepa 1-6小鼠MIP-1α(mMIP-1α),筛选并扩增抗性克隆,以流式细胞术检测m4-1BB L的表达,绘制并比较mMIP-1α和m4-1BB L同时或单独表达的Hepa 1-6细胞的生长曲线.C57B/L小鼠随机分为7组,每组9只,各组分别接种Hepa 1-6 mMIP-1α+m4-1BB L、Hepa 1-6 m4-1BB L、Hepa 1-6 mMIP-1α、Hepa 1-6 pBabe puro、Hepa 1-6、Hepa 1-6 pLXSHD和PBS,观察比较各组肝癌细胞的致瘤性,比较各组小鼠的存活率.结果 成功获得同时表达mMIP-1α和m4-1BB L的小鼠肝癌细胞Hepa 1-6 mMIP-1α+m4-1BB L,mMIP-1α和m4-1BB L同时或单独表达不影响Hepa 1-6的生长曲线.观察5周,Hepa 1-6 mMIP-1α+m4-1BB L接种的小鼠均未生长肿瘤,Hepa 1-6 mMIP-1α+m4-1BB L的体内致瘤性低于Hepa 1-6 mMIP-1α和Hepa 1-6 m4-1BB L.接种Hepa 1-6 mMIP-1α+m4-1BBL的小鼠12周末存活率(9/9)高于接种Hepa 1-6 m4-1 BB L小鼠(6/9)和Hepa 1-6 mMIP-1α小鼠(1/9).结论 趋化因子MIP-1α联合共刺激分子4-1BB L降低了肝癌细胞体内致瘤性,并使小鼠的生存期延长.

Abstract：

Objective To investigate the effects of macrophage inflammatory protein-1α (MIP-1α) combined with molecule 4-1BB L on the tumorigenicity of hepatocellular carcinoma cells in vivo. Methods Mouse MIP-1α (mMIP-1α) expressed Hepa 1-6 cells were transfected with m4-1BBL recombinant retrovirus, the anti-histidinol cells clones were selected and amplified. The expression of m4-1BB L was confirmed by flow cytometry. The growth curve of Hepa 1-6 cells transfected with mMIP-1α and m4-1BBL alone or together was drawn and compared. C57B/L Mice were randomly divided into 7 groups, 9 mice in each group, injected with mMIP-1α+m4-1BB L Hepa 1-6 cells, m4-1BB L Hepa 1-6 cells, mMIP-1α Hepa 1-6 cells, Hepa 1-6 cells, pLXSHD Hepa 1-6 cells or PBS respectively. The tumorigenicity of hepatocellular carcinoma cells and the mice survival rate were compared between each groups. Results Hepa 1-6 mMIP-1α+m4-1BB L cells which expressed both mMIP-1α and m4-1BB L were successfully established. The expression of mMIP-1α and m4-1BB L alone or together did not affect the growth curve of Hepa 1-6 cells. Observed for 5 weeks, no tumor developed in Hepa 1-6 mMIP-1α+m4-1BB L injected mice. The tumorigenicity of Hepa 1-6 mMIP-1α+m4-1BB L was lower than that of Hepa 1-6 mMIP-1α or Hepa 1-6 m4-1BB L in vivo. The survival rate of Hepa 1-6 mMIP-1α+m4-1BBL injected mice(9/9) was higher than that of Hepa 1-6 m4-1BB L injected mice (6/9)or Hepa 1-6 mMIP-1α injected mice (1/9). Conclusion Chemokine MIP-1α combined with costimulatory 4-1BB L lowered the tumorigenicity of hepatocellular carcinoma cells in vivo, and prolonged the mice survival period.     

B7-1 and 4-1BB ligand expression on a myeloma cell line makes it possible to expand autologous tumor-specific cytotoxic T cells in vitro

OBJECTIVE: The aim of this study was to confer an antigen-presenting cell (APC) ability on multiple myeloma cell lines (HMCLs) using B7-1 and/or 4-1BBL gene transfer. MATERIALS AND METHODS: HMCLs were retrovirally transduced with B7-1 and/or 4-1BBL cDNAs. Allogeneic or autologous T cells were stimulated by coculture with B7-1- and/or 4-1BBL-transduced HMCLs in the presence of interleukin-2. T cell clones were obtained by limiting dilution. T-cell activation was assessed by interferon-gamma Elispot assays and cytotoxicity by (51)Cr release assays. RESULTS: Neither primary multiple myeloma cells (MMCs) nor HMCLs expressed B7-1 or 4-1BBL, and these molecules could not be induced by CD40 triggering. HMCLs failed to stimulate allogeneic or autologous T cells. Transduction of HMCLs with B7-1 and/or 4-1BBL retroviruses induced a high expression of B7-1 and 4-1BBL molecules and a strong T-cell activation ability. Long-term cultured CD8(+) T-cell lines could be obtained by stimulation with the autologous B7-1/4-1BBL XG-19 HMCL. These cytotoxic T lymphocytes (CTL) efficiently killed the autologous parental XG-19 HMCL as well as autologous primary MMCs and allogeneic HMCLs. They did not kill autologous CD34 cells and autologous EBV cell line or natural killer target K562 cells. Cloned CTL could recognize allogeneic HMCLs, demonstrating that a shared anti-MMC repertoire was expanded. CONCLUSION: Transduction with B7-1 and 4-1BBL retroviruses turned HMCLs into efficient APCs. It permitted the long-term expansion of autologous anti-tumor CTL with a shared anti-MMC repertoire, for one HMCL. These data suggest developing an immunotherapy using modified tumor cells in patients with multiple myeloma.     

质粒介导的血管抑制和免疫强化协同诱导肝癌细胞凋亡的研究

目的 探讨真核表达质粒介导的血管抑制和免疫强化治疗诱导小鼠种植性肝癌肿瘤细胞凋亡的效果和机制.方法 制备、纯化小鼠内皮抑制素真核表达质粒(pSecES)和小鼠白细胞介素12(IL-12)质粒(pmlL-12).小鼠股部肌肉内接种H22细胞建立肝癌种植瘤模型,分成4组,接种部位分别多次注射pSecES、pmIL-12、pSecES+pmlL-12和pcDNA3.1质粒裸DNA,观察种植瘤的形成和重量;肿瘤组织切片CD31免疫组织化学染色检测肿瘤微血管密度,HE染色观察肿瘤浸润淋巴细胞,并用TUNEL检测肿瘤细胞的凋亡.结果 与pcDNA3.1对照组相比,注射pSecES,pmIL-12组小鼠肝癌种植瘤生长缓慢,血管减少,后者并可见瘤内大量淋巴细胞浸润.两组肿瘤细胞凋亡增加,pSecES、pmIL-12和pcDNA3.1组凋亡细胞数分别为(11.3±4.1)、(14.6±3.2),(1.4±1.3)个/高倍视野,pSecES+pmIL-12组凋亡细胞数为(19.9±5.5)个/高倍视野,明显高于单一治疗组.结论 真核表达质粒介导的血管抑制和免疫强化治疗可通过抑制肿瘤血管形成,促进瘤内淋巴细胞浸润,协同诱导肿瘤细胞凋亡,较好的抑制了小鼠种植性肝癌的生长.     

HIV/AIDS患者外周血程序性死亡受体-1的表达及临床意义

目的 探讨HIV感染及抗病毒治疗对程序性死亡受体-1(programmed death-1, PD-1)表达的影响。方法 根据是否接受抗病毒治疗将61例HIV/AIDS患者分为治疗组及未治疗组,并以35例健康者作为对照。利用逆转录聚合酶链反应(RT-PCR)技术探究基因PD-1 mRNA在人外周血单个核细胞(PBMC)中的表达;通过双夹心抗体ELISA法测定血清中可溶性PD-1(sPD-1)表达水平。并比较不同CD4⁺ T淋巴细胞数的HIV/AIDS患者血清sPD-1的差异。结果 未治疗组、治疗组、健康组研究对象PBMC中PD-1 mRNA的相对表达量均数分别为0.337 8±0.064、0.578 2±0.073和0.771 5±0.124,健康组与未治疗组差异极显著,健康组与治疗组、治疗组与未治疗组之间差异显著(P=0.031、P=0.043);未治疗组、治疗组、健康组血清sPD-1浓度分别为42.22±2.21 ng/mL、38.24±2.79 ng/mL和29.88±1.41 ng/mL。健康组与未治疗组、治疗组,治疗组与未治疗组分别具有显著性差异(P=0.008、P=0.040和P=0.020)。差异性分析结果表明,未治疗组、治疗组CD4⁺T淋巴细胞数<350 个/mm³患者血清sPD-1水平均显著高于CD4⁺T淋巴细胞数>350 个/mm³患者。结论 抗病毒治疗在一定程度上使高水平sPD-1表达下调以促使PD-1/PD-L1通路的恢复,从而促进机体免疫重建。监测PD-1 mRNA及sPD-1的表达在HIV的辅助诊断和推断病情发展上具有一定的价值。     

共刺激分子4-1BBL在过敏性支气管哮喘患者外周血的表达及其意义

目的 4-1BBL(CD137L)为肿瘤坏死因子超家族成员,4-1BBL与其受体4-1BB结合为T细胞活化提供共刺激信号.有研究证实激动性4-1BBL抗体能降低过敏性哮喘小鼠的气道高反应性,减少气道炎症细胞浸润.但4-1BBL在过敏性哮喘患者中的作用尚不清楚.方法 我们比较了过敏性哮喘患者和健康对照组外周血单个核细胞中...