白介素-32 基因多态性及血清水平与多发性硬化遗传易感性的关联性

目的:探讨IL-32 基因rs28372698A/ T、rs12934561C/ T 及rs11861531C/ T 三个位点的多态性与多发性硬化(MS)的遗传易感的关系,为MS 高危人群的确立提供理论依据。方法:入选580 例MS 患者和650 例健康对照,应用单碱基延伸法和DNA 测序对IL-32 基因位点进行基因分型,同时,采用酶联免疫吸附试验检测两组IL-32 的血清浓度。结果:IL-32 基因rs28372698A/ T 位点的基因型频率和对照组比较存在显著差异(P =0.007),其等位基因频率在两组间的分布频率存在统计差异(P =0.033)。rs12934561C/ T 与rs11861531C/ T 的各基因型及等位基因频率在两组间差异无统计学意义(P>0.05)。T-T-T单倍型在HCC 中的分布频率显著高于对照组(P = 0.012),T-T-T 单倍型与MS 的发病风险密切相关(OR = 1.968,95% CI:1.352-2.574)。MS 患者组的血清IL-32 水平明显高于对照组[(399.08±156.85)pg/ ml vs(239.99±88.35)pg/ ml,P =0.001]。AT 和TT 基因型的MS 患者IL-32 血清水平明显高于AA 基因型MS 患者[(465.53 ±172.40) pg/ mL vs (295.86 ±103.96)pg/ ml,P<0.01;(491.15±133.65)pg/ ml vs (295.86±103.96)pg/ ml,P<0.01]。结论:本研究首次报道了IL-32 基因rs28372698 位点多态性与MS 的关系,且IL-32 的基因多态性在MS 患者中对IL-32 的血清水平有影响。我们的研究为MS 遗传和个体化的诊疗提供了新的参考依据。     

血清中IL-17 水平及IL-17rs763780 基因多态性与深圳地区Graves 病患者遗传易感性分析

目的:了解Graves病患者血清中IL-17水平及IL-17rs763780基因多态性分布情况,并探讨IL-17rs763780基因多态性与深圳地区Graves病患者之间的遗传基因易感性。方法:收集2016年2月～2018年11月来深圳市大鹏新区妇幼保健院门诊或/和住院部就诊并确诊为Graves病患者152例为Graves组,选取同期来医院体检中心体检的健康人群100名为对照组,采用酶联免疫吸附法(ELISA)检测血清中IL-17水平,同时采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)法对IL-17rs763780基因多态性进行检测,并对检测结果进行统计分析。结果:Graves组患者血清中IL-17水平为(495.47±190.52)pg/ml,明显高于对照组的(164.13±42.95)pg/ml,差异有统计学意义(t=10.627 2,P0.05);男性Graves病患者血清中IL-17水平为(427.26±152.67)pg/ml,低于女性患者的(561.73±201.63)pg/ml,差异有统计学意义(t=2.764 5,P0.05);Graves病组IL-17rs763780 CC基因型及C等位基因频率分别为20.39%和34.87%,明显高于对照组,差异有统计学意义(χ~2=2.810 2～3.275 6,P0.05);男性Graves病患者IL-17rs763780 CC基因型及C等位基因频率分别为16.42%和29.10%,低于女性患者,差异均有统计学意义(χ~2=2.038 4～2.615 3,P0.05);IL-17rs763780 CC基因型患者IL-17水平为(549.24±197.25)pg/ml,略高于TC基因型的(524.29±193.87)pg/ml,差异无统计学意义(t=0.941 4,P0.05),但两者均明显高于TT基因型患者的(371.83±162.96)pg/ml,差异有统计学意义(t=2.502 1～2.871 0,P0.05)。结论:Graves病患者血清中IL-17水平明显增高,IL-17rs763780 CC基因型及C等位基因频率明显高于健康人群,CC和TC基因型患者血清中IL-17水平高于TT基因型。因此,IL-17rs763780 CC基因型及C等位基因可能是导致深圳地区Graves发病的危险遗传基因之一。     

毛细支气管炎患儿血清IL-4,IgE测定及与哮喘的相关性研究

目的观察毛细支气管炎患儿血清IL-4,IgE的变化.方法测定36例支气管哮喘发作期,42例毛细支气管炎,36例支气管肺炎患儿血清IL-4,IgE水平.结果毛细支气管炎患儿血清IL-4,IgE水平(分别为141.67±44.97pg/ml,124.76±50.45IU/ml)介于哮喘组(分别为19I.64±38.87pg/ml,199.87±65.63IU/ml)与肺炎组(分别为55.44±27.96pg/ml,79.01±69.28IU/ml)之间,有显著性差异(P均＜0.01).结论毛细支气管炎和哮喘可能存在着相同或相似的免疫学发病机制.     

Mina和IL-4基因交互作用与儿童哮喘相关性研究

目的研究Mina基因rs4857304(G/T)位点、IL-4基因位点rs2243248(T/G)和rs2070874(T/C)单核苷酸多态性与中国汉族儿童过敏性哮喘发病相关性,同时对3个基因位点之间交互作用进行分析。方法采用病例对照研究,MALDI-TOF技术进行IL-4和Mina位点基因分型。观察不同基因型和等位基因频率在哮喘和对照组中的分布差异,用MDR软件分析各位点之间交互作用。结果共纳入哮喘患儿202例年龄在6.97±2.85岁之间,对照组191例年龄在8.14±3.13岁之间。Mina的rs4857304位点,在哮喘组和对照组等位基因频率比较差异具有统计学意义(P=0.0199,OR=1.477,95%CI=1.062~2.052),哮喘组rs4857304位点等位基因T频率高于对照组,而两组之间该位点基因型分布比较差异无统计学意义(P<0.05)。IL-4基因位点rs2243248和rs2070874基因型和等位基因频率在两组比较差异均无统计学意义(P>0.05)。MDR交互作用分析显示,IL-4基因位点rs2243248和Mina基因位点rs4857304构成的2个位点最佳模型;IL-4基因位点rs2243248、rs2070874和Mina位点rs4857304构成的3个位点最佳模型两组比较均有统计学差异(P<0.05)。结论 Mina基因位点rs4857304与过敏性哮喘具有相关性,同时IL-4位点rs2243248,rs2070874与Mina rs4857304均具有明显交互作用。     

类风湿关节炎患者血清中IL-17 表达及IL-17F 基因位点rs763780 多态性分析

目的:了解类风湿关节炎(RA)患者血清中IL-17、类风湿因子(RF)及C反应蛋白(CRP)的表达,并探讨IL-17F基因位点rs763780多态性与RA发病易感相关性。方法:收集2016年1月～2018年10月来深圳市福田区风湿病专科医院就诊的RA患者176例为研究组,并选取同期来院体检的健康人群110名为对照组,分别检测血清中IL-17、RF及CRP含量,并采用聚合酶链反应(PCR-RFLP)技术对IL-17F基因位点rs763780多态性进行分析。结果:RA组血清中IL-17、RF及CRP含量分别为(36.73±11.06)pg/ml,(58.05±14.92)U/ml和(32.15±13.96)mg/L,明显高于对照组的(14.52±6.35)pg/ml,(10.24±5.32)U/ml和(6.81±3.25)mg/L,差异均有统计学意义(t=3.035 7～5.320 6,P0.05);RA组CC基因型和C等位基因检出率分别为19.32%和41.48%,明显高于对照组的6.36%和26.82%,差异有统计学意义(χ~2=3.916 5～5.054 7,P0.05);RA组IL-17F基因位点rs763780 CC基因型中的IL-17水平为(42.96±13.64)pg/ml,明显高于其他基因型,差异有统计学意义(P0.05),而三种基因型血清中RF和CRP水平之间差异均无统计学意义(P0.05)。结论:RA患者血清中IL-17,RF及CRP水平明显高于健康人群,同时RA患者IL-17F基因位点rs763780 CC基因型及C等位基因检测率明显高于健康人群,可能是导致本地区RA发病的危险遗传易感基因之一。     

兰州地区IL-15、IL-17A和IL-18基因多态性与复发性流产的相关性研究北大核心CSCD

目的:探讨IL-15、IL-17A和IL-18基因多态性与复发性流产(RSA)的关系。方法:SNaPshot法分析150例RSA患者(RSA组)和150例健康志愿者(对照组)中IL-15基因rs3806798、IL-17A基因rs2275913、IL-18基因rs1946518和rs187238位点多态性分布情况。结果:RSA组和对照组中4个单核苷酸多态性(SNPs)基因型频率和等位基因频率分布在两组间差异无统计学意义(P>0.05);隐性和显性遗传模式下对IL-15基因rs3806798、IL-17A基因rs2275913、IL-18基因rs1946518进行非条件Logistic回归分析,发现两组间差异均无统计学意义(P>0.05)。但IL-18基因的rs187238 SNP位点GG型、CG型、CC型3种基因型构成比在两组间差异有统计学意义(χ2=9.256,P=0.010)。显性、隐性遗传模式Logistic回归分析结果显示G等位型(GC+GG基因型)与RSA患病风险降低相关(χ2=4.303,OR=0.53,95%CI=0.29~0.97,P=0.038),但G等位基因不是RSA的保护性等位基因(χ2=2.275,OR=0.66,95%CI=0.38~1.14,P=0.132)。经连锁不平衡和单体型分析,rs1946518和rs187238存在完全连锁不平衡。TGTG单体型是RSA的保护因素,其OR(95%CI)=0.361(0.158~0.827)。结论:IL-18基因rs187238基因多态性与RSA患病风险相关。     

IL-17、IL-33水平及IL-33基因rs16924159G/A位点多态性与不明原因复发性流产易感性分析

目的了解深圳地区不明原因复发性流产(unexplained recurrent spontaneous abortion,URSA)患者血清中白细胞介素-17(interleukin-17,IL-17)、IL-33水平及IL-33基因rs16924159G/A位点多态性分布情况,并探讨其与URSA发病的易感性。方法收集2019年2月～2020年1月来医院妇科门诊就诊并确诊为URSA患者132例为URSA组,并选择同期来医院妇科门诊要求人工流产的早孕女性120名为对照组,采用酶联免疫吸取附法(ELISA)检测血清中IL-17和IL-33水平,同时采用多重连接酶检测反应(improved multi-ligase detection reaction,imLDR)技术检测IL-33基因rs16924159G/A位点多态性。结果 URSA患者血清中IL-17水平为56.01±19.43ng/L,明显高于对照组的32.85±11.27ng/L,而IL-33水平为105.97±26.32pg/ml,明显低于对照组的154.68±34.76pg/ml,差异均有统计学意义(P0.05);URSA患者IL-33基因rs16924159G/A位点AA基因型和A等位基因检出率分别为28.03%和44.94%,明显高于对照组的9.17%和23.75%,差异均有统计学意义(P0.05);携带IL-33基因rs16924159G/A位点AA基因型URSA患者血清中IL-33水平为76.08±19.25pg/ml,明显低于GG和GA基因型的121.52±30.67pg/ml和113.71±28.06pg/ml,差异有统计学意义(P0.05),而GG和GA基因型之间差异无统计学意义(P0.05),同时对照组中不同基因型血清中IL-33水平之间差异均无统计学意义(P0.05);经Spearman相关性分析,URSA患者血清中IL-17和IL-33水平呈明显负相关(r=-0.6706,P0.05)。结论 URSA发病可能与IL-33水平降低,其对Th17细胞分泌抑制作用减弱有关,而IL-33水平降低可能与其基因rs16924159G/A位点多态性突变有关,其中AA基因型可能是URSA发病的危险易感基因之一。     

IL-18 水平表达及IL-18 基因多态性与哮喘病的相关性研究

目的:探讨IL-18基因多态性位点(IL-18/-607C/A,IL-18/-137G/C)多态性及血清IL-18水平与哮喘发生的关系。方法:选取我院301例哮喘患者作为哮喘组,并选取288例健康成人作为健康组。提取研究对象全血DNA,采用等位基因特异性引物PCR检测两组IL-18基因多态性位点(IL-18/-607C/A,IL-18/-137G/C),并对其PCR反应产物进行测序验证;观察IL-18基因多态性位点(IL-18/-607C/A,IL-18/-137G/C)等位基因频率分布,同时通过酶联免疫吸附法检测IL-18在不同组别血清浓度,分析影响哮喘发生的危险因素。结果:与对照组相比IL-18基因(-607C/A)、3种基因型、等位基因等差异较大(χ2=10. 24,P0. 001;χ2=50. 26,P0. 001),差异具有显著统计学意义。哮喘组基因位点-137G/C的CC与GG基因型差异有显著统计学意义(χ2=4. 717,P0. 05),其等位基因频率差异无显著性(χ2=3. 711,P0. 05);哮喘组患者血清IL-18水平显著低于健康组(t=85. 34,P0. 001),其中基因多态性位点(-607C/A) CC基因型的哮喘患者血清IL-18的水平为(18. 02±3. 92) pg/ml,携带AA基因型患者IL-18的水平为(41. 68±8. 08) pg/ml,两者比较差异具有统计学意义(t=22. 26,P0. 001);基因多态性位点(-137G/C) 3种基因型哮喘患者血清IL-18的水平差异无统计学意义(F=0. 281,P0. 05)。结论:哮喘患者血清IL-18水平降低,提示哮喘的发生可能与血清IL-18水平低下有关。IL-18多态性位点(IL-18/-607C/A,IL-18/-137G/C)的基因分型中携带(-607C/A) AA基因型的人群患哮喘风险更大。     

OX40L基因多态性与复发性流产相关性的研究

目的通过检测OX40L基因rs1234315、rs2205960、rs17568、rs2298212位点多态性基因频率,探讨这些位点与复发性自然流产之间的关系。方法采用EILSA方法检测65例复发性自然流产患者以及75例正常妊娠患者外周血血清IL-10、IL-2细胞因子;采用直接测序法检测上述患者OX40L基因rs1234315、rs2205960、rs17568、rs2298212等位点多态性。结果复发性自然流产组患者血清中IL-2水平(83.82±50.29pg/m L)显著高于正常妊娠组(66.42±30.64pg/m L)(P0.05);复发性自然流产组患者血清中IL-10(18.86±12.63pg/m L)显著低于正常妊娠组(24.96±9.42pg/m L)(P0.05)。两组OX40L基因rs1234315、rs2205960、rs17568多态性位点的基因型和等位基因频率均无统计学差异(P0.05):复发性自然流产组rs2298212基因A/A基因型频率显示高于正常妊娠组(P0.05),两组G/G基因型与G/A基因型无显著差异(P0.05)。结论 OX40L基因SNPrs2298212基因A/A基因型可能是复发性自然流产危险因素,需要进一步研究证实。     

哮喘儿童血清白细胞介素-25、嗜酸细胞趋化因子的动态变化和意义

目的:通过监测哮喘儿童血清白细胞介素(IL)-25、嗜酸细胞趋化因子(Eotaxin)表达水平, 探讨其水平变化及其在哮喘发病中的作用.方法:收集哮喘急性发作期、缓解期儿童及健康儿童的静脉血标本, 用双抗体夹心ELISA法测定血清IL-25、Eotaxin的水平.结果:哮喘儿童急性发作期组血清IL-25水平(56.75±11.68) ng/L显著高于缓解期组(47.09±10.96) ng/L及健康对照组(45.77±10.43) ng/L, 差异有统计学意义(P＜0.05), 缓解期组与健康对照组比较差异无统计学意义;哮喘急性发作期组儿童血清Eotaxin水平(120.95±23.97) ng/L显著高于缓解期组(105.47±22.01) ng/L及健康对照组(105.01±18.47) ng/L , 差异有统计学意义(P＜0.05), 缓解期组与健康对照组比较差异无统计学意义.哮喘儿童血清IL-25水平与Eotaxin之间存在明显正相关关系(r=0.642, P＜0.01).结论:哮喘儿童血清IL-25及Eotaxin 水平能反应哮喘气道炎症活动情况及疾病严重程度, 两者存在正相关关系.     

The extended IL-10 superfamily: IL-10, IL-19, IL-20, IL-22, IL-24, IL-26, IL-28, and IL-29

Cytokines are involved in virtually every aspect of immunity and inflammation. A cascade of responses evolves after cytokine activation, although optimal function might ultimately involve several complementary cytokines. Understanding the function of individual cytokines is complicated because their role can vary depending on the cellular source, target, and phase of the immune response. In fact, numerous cytokines have both proinflammatory and anti-inflammatory potential, with the contrasting outcome observed being determined by the immune cells present and their state of responsiveness to the cytokine. These issues make the study of cytokine biology daunting, particularly so for IL-10 and IL-10-related genes. The IL-10 superfamily is highly pleiotropic. These genes are linked together through genetic similarity and intron-exon gene structure. Significant commonality exists not only through shared receptors but also through conserved signaling cascades. However, its members mediate diverse activities, including immune suppression, enhanced antibacterial and antiviral immunity, antitumor activity, and promotion of self-tolerance in autoimmune diseases.     

IL-10 subfamily members: IL-19, IL-20, IL-22, IL-24 and IL-26

It has been reported that the CD4+ T cell is a very important source of interleukin 10 (IL-10), while CD8+ cells produce low amounts. IL-10 exerts several immune stimulating, as well as inhibitory effects. There are at least five novel human IL-10 family-related molecules: IL-19, IL-20, IL-22, IL-24, and IL-26. Activated T cells produce IL-19, IL-22 and IL-26, while IL-24 is produced by activated monocytes and T-cells. IL-20 induces cheratin proliferation and Stat-3 signal transduction pathway, while IL-22 induces acute-phase production by hepatocytes and neonatal lethality with skin abnormalities reminiscent of psoriasic lesions in humans. In addition, IL-22 mediates inflammation and binds class II cytokine receptor heterodimers IL-22 RA1/CRF2-4. This cytokine is also involved in immuno-regulatory responses. IL-26 (AK155) is a novel cytokine generated by memory cells and is involved in the transformed phenotype of human T cells after infection by herpes virus. All these new IL-10 subfamily member cytokines are strongly involved in immune regulation and inflammatory responses.     

IL-21 and IL-21 receptor

Interleukin (IL)-21 is a new member of the type I cytokine superfamily. Although it is most homologous to IL-15, it has a unique receptor chain, IL-21R, that pairs with the γ-common cytokine receptor chain. The first experiments examining the biology of the IL-21 pathway reveal that it is a cytokine with effects on natural killer (NK) cells, T cells, and B cells. Mice deficient in the IL-21 R have also been made, and are being examined for the effects of the IL-21/IL-21R pathway in vivo. Here we summarize our current knowledge of this new cytokine pathway, and its role in innate and adaptive immunity.     

Ovarian follicular concentration of IL-12, IL-15, IL-18 and p40 subunit of IL-12 and IL-23

BACKGROUND: The aim of the study was to determine the presence of interleukin (IL)-12, IL-15, IL-18 and p40 subunit of IL-12/IL-23 in follicular fluid from spontaneous cycles and the relation between the concentration of selected cytokines and IVF-embryo transfer outcome. METHODS: IVF-embryo transfer and enzyme immunoassay (EIA) (R&D Systems, Minneapolis, MN, USA and MBL, Nagoya, Japan) were used. RESULTS: Follicular fluid of women included in the IVF-embryo transfer procedure contained common p40 subunit of IL-12/IL-23 (median 70.1 pg/ml), IL-15 (median 1.3 pg/ml) and IL-18 (median 38.2 pg/ml). There was a significant negative correlation between follicular fluid concentrations of IL-15 and IL-18 (R=-0.392, P=0.003). Significantly higher concentrations of common p40 subunit of IL-12/IL-23 (median 79.8 pg/ml) were found in the follicular fluid taken from follicles containing oocytes, when compared with those without an oocyte (median 44.5 pg/ml, P=0.006). Patients who achieved clinical pregnancy had significantly decreased concentration of IL-15 (median 0.8 pg/ml) compared with patients without successful IVF-embryo transfer outcome (median 1.4 pg/ml, P=0.047). CONCLUSION: Follicular fluid collected from spontaneous cycles contains detectable levels of p40 subunit of IL-12/IL-23, IL-15 and IL-18. Increased concentrations of p40 subunit of IL-12/IL-23 in follicles containing oocytes suggest an important role of this cytokine in reproduction. Possible negative value of IL-15 as a predictor of IVF-embryo transfer success remains to be determined.     

A tissue specific IL-1 receptor antagonist homolog from the IL-1 cluster lacks IL-1, IL-1ra, IL-18 and IL-18 antagonist activities

Interleukin (IL)-1-like protein 1 (IL-1L1) is a 155-amino acid protein that shares 27% identity with IL-1beta and 47% with IL-1 receptor antagonist (IL-1ra). A 2.7-kb IL-1L1 mRNA was cloned from human placenta and is detectable in the trophoblastic cell line JEG-3, in macrophages and in endotoxin-stimulated monocytes. Expression of IL-1L1 is much less abundant and less widespread than IL-1ra. We have determined the human and mouse IL-1L1 cDNA sequences and the complete sequence of the human gene, IL1L1. IL1L1 consists of four coding exons, has two alternative non-coding first exons, lies between IL1B and IL1RN, is orientated in the same direction as IL1RN and is separated from it by approximately 53 kb. The predicted IL-1L1 protein lacks both signal sequence and glycosylation signals. A 17-kDa protein was recovered by immunoprecipitation with IL-1L1-specific antibodies from JEG-3. IL-1L1 did not stimulate IL-6 production from primary human fibroblasts or human umbilical vein endothelial cells nor did it block the IL-1alpha or IL-1beta-dependent activation of IL-6 expression. We conclude, contrary to a recent suggestion made by others, that IL-1L1 is not a functional IL-1ra. IL-1L1 also had no detectable agonistic or antagonistic effect on IFN-gamma production in response to IL-18 in KG-1 cells.     

IL-1, IL-18, and IL-33 families of cytokines

Summary: The interleukin-1 (IL-1), IL-18, and IL-33 families of cytokines are related by mechanism of origin, receptor structure, and signal transduction pathways utilized. All three cytokines are synthesized as precursor molecules and cleaved by the enzyme caspase-1 before or during release from the cell. The NALP-3 inflammasome is of crucial importance in generating active caspase-1. The IL-1 family contains two agonists, IL-1α and IL-1β, a specific inhibitor, IL-1 receptor antagonist (IL-1Ra), and two receptors, the biologically active type IL-1R and inactive type II IL-1R. Both IL-1RI and IL-33R utilize the same interacting accessory protein (IL-1RAcP). The balance between IL-1 and IL-1Ra is important in preventing disease in various organs, and excess production of IL-1 has been implicated in many human diseases. The IL-18 family also contains a specific inhibitor, the IL-18-binding protein (IL-18BP), which binds IL-18 in the fluid phase. The IL-18 receptor is similar to the IL-1 receptor complex, including a single ligand-binding chain and a different interacting accessory protein. IL-18 provides an important link between the innate and adaptive immune responses. Newly described IL-33 binds to the orphan IL-1 family receptor T1/ST2 and stimulates T-helper 2 responses as well as mast cells.     

支气管哮喘病人胸导管淋巴液与血清IL-6、IL-8、 IL-10、IL-12、TNF-α水平的对比研究

目的：探讨哮喘病人胸导管淋巴液和血清IL-6、IL-8、IL-10、IL-12及TNF-α水平变化。方法：采用酶联免疫吸附试验（ELISA）检测31例行胸导管引流治疗的中、重度哮喘病人术后0 d、3 d、5 d淋巴液及0 d、5 d血清IL-6、IL-8、IL-10、IL-12以及TNF-α水平。结果：哮喘病人胸导管引流淋巴液中IL-6、IL-8、IL-10、TNF-α水平均高于正常对照组血清水平，而IL-12则明显低于正常血清水平；引流5 d淋巴液中IL-6、IL-8、IL-10及TNF-α明显低于，IL-12则显著高于引流0 d时水平；同时哮喘病人血清IL-10、TNF-α含量低于，血清IL-12则高达正常对照组水平。结论：哮喘病人淋巴液中存在以IL-12产生受抑和炎性细胞因子产生亢进为特征的细胞因子失调，且淋巴液中CK变化与血清变化大致平行。     

IL-4、IL-13蛋白表达及抗IL-4、IL-13人源单链抗体的筛选

目的:表达IL-4和IL-13蛋白,从人源单链抗体文库中分别筛选抗IL-4和抗IL-13单链抗体.方法:采用RT-PCR从健康志愿者外周血单核细胞(PBMC) mRNA中扩增IL-4和IL-13 cDNA;构建硫氧还蛋白融合表达载体,转化大肠杆菌BL21,IPTG诱导表达并对表达产物进行纯化鉴定.以生物素化的IL-4和IL-13为抗原从前期构建的人源抗体文库中采用噬菌体展示技术分别筛选抗IL-4和抗IL-13人源单链抗体(scFv).结果:扩增的IL-4 cDNA大小为280 bp,表达的融合蛋白大小为27 kD左右.扩增的IL-13 cDNA大小为252 bp,表达的融合蛋白大小为25 kD左右.分别以生物素化的IL-4和IL-13蛋白为抗原,采用噬菌体展示技术对人源抗体文库进行3轮富集后,分别有大约37％的scFvs与IL-4有结合特性,有约27％的scFvs与IL-13有结合特性.筛选了4株分别与IL-4和IL-13结合能力强的单链抗体进行了Westem blot鉴定和测序.结论:成功筛选到抗IL-4和抗IL-13人源性单链抗体.     

Proinflammatory cytokines (IL-17, IL-6, IL-18 and IL-12) and Th cytokines (IFN-gamma, IL-4, IL-10 and IL-13) in patients with allergic asthma

Allergen-reactive T helper type-2 (Th2) cells and proinflammatory cytokines have been suggested to play an important role in the induction and maintenance of the inflammatory cascade in allergic asthma. We compared the plasma concentrations of novel proinflammatory cytokines IL-17 and IL-18, other proinflammatory cytokines IL-6 and IL-12, Th2 cytokines IL-10 and IL-13, and intracellular interferon-gamma (IFN-gamma) and IL-4 in Th cells of 41 allergic asthmatics and 30 sex- and age-matched health control subjects. Plasma cytokines were measured by enzyme-linked immunosorbent assay. Intracellular cytokines were quantified by flow cytometry. Plasma IL-18, IL-12, IL-10, IL-13 concentrations were significantly higher in allergic asthmatic patients than normal control subjects (IL-18: median 228.35 versus 138.72 pg/ml, P < 0.001; IL-12: 0.00 versus 0.00 pg/ml, P = 0.001; IL-10: 2.51 versus 0.05 pg/ml, P < 0.034; IL-13: 119.38 versus 17.89 pg/ml, P < 0.001). Allergic asthmatic patients showed higher plasma IL-17 and IL-6 concentrations than normal controls (22.40 versus 11.86 pg/ml and 3.42 versus 0.61 pg/ml, respectively), although the differences were not statistically significant (P = 0.077 and 0.053, respectively). The percentage of IFN-gamma-producing Th cells was significantly higher in normal control subjects than asthmatic patients (23.46 versus 5.72%, P < 0.001) but the percentage of IL-4 producing Th cells did not differ (0.72 versus 0.79%, P > 0.05). Consequently, the Th1/Th2 cell ratio was significantly higher in normal subjects than asthmatic patients (29.6 versus 8.38%, P < 0.001). We propose that allergic asthma is characterized by an elevation of both proinflammatory and Th2 cytokines. The significantly lower ratio of Th1/Th2 cells confirms a predominance of Th2 cells response in allergic asthma.     

Polymorphisms of IL-1B, IL-1RN, IL-2, IL-4, IL-6, IL-10, and IFN-gamma genes in the Korean population

Cytokines play a crucial role in regulating the immune and inflammatory responses. The collective influence of several cytokines can regulate immune responses as complex as those underlying allograft rejections or autoimmune diseases. Polymorphisms in the regulatory regions of the cytokine genes may influence their expression. Therefore, the polymorphisms of cytokine genes are potentially important as genetic predictors of the disease susceptibility or clinical outcome. In 311 unrelated healthy Korean individuals, we investigated the polymorphisms of cytokine genes (interleukin-1 [IL-1], IL-2, IL-4, IL-6, IL-10, and interferon-gamma [IFN-gamma]), which had been previously reported to be associated with a number of immune diseases, transplant complications, and direct or indirect influences on the level of expression and production. And we also compared the results to those published for other populations. The genotype distributions were consistent with the assumption of the Hardy-Weinberg equilibrium, with the exceptions of IL-1B +3954 and IL-6-174 polymorphisms. The polymorphisms examined in this study were almost similar to that observed in Asian populations. There were significant differences of the polymorphisms, except for IL-4 receptor alpha +1902, between Korean and other populations. Comparing the alleles associated with higher level of expression and production, IL-1B +3954*T, IL-2-330*G, and IL-4-590*T alleles were significantly higher, and IL-1RN*A2, IL-10-1082*G, and IFN-gamma*2 alleles were lower in Koreans than other populations. Especially in IL-6 promoter -174 polymorphism, we found only the G allele associated with higher plasma IL-6 levels. In haplotype analysis of IL-10 promoter polymorphisms, the GCC haplotype, associated with higher expression of IL-10, was significantly lower in Koreans. These results may be helpful for understanding transplant-related complications, immune or autoimmune diseases, and malignant diseases in the Korean population.