一对高亲和力抗人肌钙蛋白Ⅰ单克隆抗体的制备及鉴定

目的 获取抗人肌钙蛋白Ⅰ（cTnI）的单克隆抗体（McAb）.方法 以人cTnI作为抗原,免疫Balb/c小鼠,通过杂交瘤技术制备了抗人cTnI高亲和力、高特异性单克隆抗体.随后采用间接ELISA法测定抗血清效价,用protein G亲和纯化法纯化抗体,抗原竞争ELISA法鉴定抗体亲和力,SDS-PAGE法鉴定纯度,Western blotting鉴定抗cTnI单克隆抗体的特异性,竞争ELISA法分析抗原结合位点.结果 筛选出9株稳定分泌抗cTnI的单抗杂交瘤细胞株,其中A3、A9两株免疫球蛋白亚类均为IgG2a,分泌的抗体纯度高,与CK-MB、cTnT无交叉反应,效价均为：1：1024000,亲和力分别为4.21×10^8mol/L、1.07×10^8mol/L,抗原结合位点不同.结论 成功制备出了一对高亲和力、高特异性抗人cTnI单克隆抗体.     

新型肿瘤标志物:Mu-GlcNAc

作者发现了一个新的肿瘤标志物MA153,其化学结构为Mu-GlcNAc（粘蛋白1-N-乙酰氨基葡萄糖）,它可与单克隆抗体Ma695和相关Aptamer（适配体MA153-A）形成Ma695-Mu-GlcNAc-MA153-A式的夹心反应。临床血清学检测表明,Mu-GlcNAc存在于癌症患者血中,以1U/mL为正常界值,Mu-GlcNAc的肿瘤特异度达95%,癌症检出的灵敏度对乳腺癌、胰腺癌、肝癌、肺癌、食管癌、口腔癌、结肠癌、胆管癌、卵巢癌、宫颈癌、子宫癌,分别为80%、74%、68%、70%、75%、50%、60%、55%、70%、64%、60%。本研究资料显示,Mu-GlcNAc是一个新的良好广谱肿瘤标志物。     

抗核抗体谱的检测在临床应用中的研究进展

自身免疫性疾病（autoimmune diseases,AID）确切的发病机制目前尚不清楚,且临床症状和体征不典型,对其诊断和鉴别诊断较复杂,但由于该类疾病都存在一些特异性的自身抗体,所以对自身免疫性疾病的诊断不仅依赖于临床表现,而且很大程度上取决于免疫学实验诊断,特别是实验室的抗核抗体（antinuclear antibody,ANA）检测[1].通过对ANA类型及其滴度的检测,能为自身免疫性疾病的诊断、分型、治疗和预后判断提供重要的依据,本文就ANA和ANA谱的检测方法及其临床应用做一综述.     

抗ProGRP(31-98) scFv的131I标记及生物分布研究

目的 研究抗胃泌素释放前体（ProGRP（31-98）单链抗体（scFv）的131I标记方法,并对其标记后的稳定性、免疫活性及生物分布进行分析.方法 采用氯胺T法碘化标记制备131I-anti-ProGRP（31-98）scFv,凝胶柱层析法分离纯化标记产物,利用纸层析法测定标记物的标记率、放化纯度和稳定性,采用细胞结合分析法比较131I-anti-ProGRP（31-98）scFv对小细胞肺癌细胞株NCI-H446和肺腺癌细胞株A549的免疫结合率.将131I-ProGRP（31-98）scFv注射入动物体内,研究其在实验动物体内的分布情况.结果 131I-anti-ProGRP（31-98）标记率为（93.35±0.67）％,标记产物纯化后即刻放化纯度为（98.49±1.21）％.131I-anti-ProGRP（31-98）scFv对人小细胞肺癌NCI-H446和肺腺癌A549细胞株的免疫结合率分别为（85.36±1.45）％和（21.02±2.16）％,且差异有统计学意义（P＜0.05）.131I-anti-ProGRP（31-98）scFv在实验动物体内主要通过肾脏和肝脏代谢,血液清除快.结论 131I-anti-ProGRP（31-98） scFv的标记率高,且有良好的稳定性和免疫活性,在实验动物体内主要通过肝脏和肾脏代谢,血液清除快.     

Induction of long-term immunity against respiratory syncytial virus glycoprotein by an osmotic polymeric nanocarrier

Respiratory syncytial virus (RSV) is one of the most common causes of viral deaths in infants worldwide, yet no effective vaccines are available. Here, we report an osmotically active polysaccharide-based polysorbitol transporter (PST) prepared from sorbitol diacrylate and low-molecular-weight polyethylenimine (PEI) showing a potent, yet safe, adjuvant activity and acting as an effective delivery tool for RSV glycoprotein (RGp) antigen. PST showed no toxicity in vitro or in vivo, unlike PEI and the well-known experimental mucosal adjuvant cholera toxin (CT). PST formed nano-sized complexes with RGp by simple mixing, without affecting antigenic stability. The complexes exhibited negative surface charges that made them highly efficient in the selective activation of phagocytic cells and enhancement of phagocytic uptake. This resulted in an improved cytokine production and in the significant augmentation of RGp-specific antibody production, which persisted for over 200 days. Interestingly, PST/RGp enhanced phagocytic uptake owing to the osmotic property of PST and its negative zeta potential, suggesting that PST could selectively stimulate phagocytic cells, thereby facilitating a long-lived antigen-specific immune response, which was presumably further enhanced by the polysaccharide properties of PST.     

Assessment of the response to gluten-free diet in an Iraqi population with coeliac disease. A histological and serological follow-up study

Introduction

Coeliac disease (CD) is a common diagnosis among children and adults in Iraq; however, removal of gluten from the diet is essential for patients with CD. The aim of this study, the first such study in Iraq, was to assess the serological and histological recovery profiles of coeliac patients, in both children and adults groups after commencing a gluten-free diet (GFD) for at least 1 year ± 1 month.

Material and methods

The study group comprised 78 proved coeliac patients (46 children and 32 adults, median age: 15 years, range: 1–66 years) who all agreed to undergo endoscopy in addition to serological assessment before and after treatment. The duodenal biopsies were interpreted histologically according to modified Marsh criteria and the sera were tested for anti-gliadin antibody (AGA), endomysium antibody (EMA) and anti-tissue transglutaminase antibody (tTG).

Results

Complete histological remission was seen in 29 (63.1%) of 46 treated children CD patients, while only 5 (10.9%) showed Marsh IIIa changes compared with 11 (24%) before GFD. Similarly none of the 32 adults after GFD showed Marsh IIIb and Marsh IIIc compared with 46.9% and 28.1% before treatment respectively (p = 001). Meanwhile, there was strongly significant reduction in AGA, EMA, and tTG antibodies levels (p = 0.00001) following GFD.

Conclusions

Repeating the duodenal biopsy 1 year ±1 month after diagnosis and starting a GFD supports the routine measurement of using histological findings as a gold standard test to confirm recovery of Iraqi CD patients along with using known coeliac serology antibodies.     

Patients co-infected with hepatitis C virus (HCV) and human immunodeficiency virus recover genotype cross-reactive neutralising antibodies to HCV during antiretroviral therapy

When severely immunodeficient HIV/HCV co-infected patients are treated with antiretroviral therapy, it is important to know whether HCV-specific antibody responses recover and whether antibody profiles predict the occurrence of HCV-associated immune restoration disease (IRD). In 50 HIV/HCV co-infected patients, we found that antibody reactivity and titres of neutralising antibodies (nAb) to JFH-1 (HCV genotype 2a virus) increased over 48 weeks of therapy. Development of HCV IRD was associated with elevated reactivity to JFH-1 before and during the first 12 weeks of therapy. Individual analyses of HCV IRD and non-HCV IRD patients revealed a lack of an association between nAb responses and HCV viral loads. These results showed that increased HCV-specific antibody levels during therapy were associated with CD4⁺ T-cell recovery. Whilst genotype cross-reactive antibody responses may identify co-infected patients at risk of developing HCV IRD, neutralising antibodies to JFH-1 were not involved in suppression of HCV replication during therapy.     

Tolerogenic dendritic cells induce antigen-specific hyporesponsiveness in insulin- and glutamic acid decarboxylase 65-autoreactive T lymphocytes from type 1 diabetic patients

Tolerogenic dendritic cells (tDC) constitute a promising therapy for autoimmune diseases, since they can anergize T lymphocytes recognizing self-antigens. Patients with type 1 diabetes mellitus (T1D) have autoreactive T cells against pancreatic islet antigens (insulin, glutamic acid decarboxylase 65 -GAD65-). We aimed to determine the ability of tDC derived from T1D patients to inactivate their insulin- and GAD65-reactive T cells. CD14 + monocytes and CD4 + CD45RA- effector/memory lymphocytes were isolated from 25 patients. Monocyte-derived DC were generated in the absence (control, cDC) or presence of IL-10 and TGF-β1 (tDC), and loaded with insulin or GAD65. DC were cultured with T lymphocytes (primary culture), and cell proliferation and cytokine secretion were determined. These lymphocytes were rechallenged with insulin-, GAD65- or candidin-pulsed cDC (secondary culture) to assess whether tDC rendered T cells hyporesponsive to further stimulation. In the primary cultures, tDC induced significant lower lymphocyte proliferation and IL-2 and IFN-γ secretion than cDC; in contrast, tDC induced higher IL-10 production. Lymphocytes from 60% of patients proliferated specifically against insulin or GAD65 (group 1), whereas 40% did not (group 2). Most patients from group 1 had controlled glycemia. The secondary cultures showed tolerance induction to insulin or GAD65 in 14 and 10 patients, respectively. A high percentage of these patients (70–80%) belonged to group 1. Importantly, tDC induced antigen-specific T-cell hyporesponsiveness, since the responses against unrelated antigens were unaffected. These results suggest that tDC therapy against multiple antigens might be useful in a subset of T1D patients.