戊型肝炎病毒ORF2N-端和C-端多肽的抗原性分析

目的分析戊型肝炎病毒(HEV)ORF2N-端和C-端多肽的抗原性。方法以纯化的4型HEVORF2N-端1～124aa多肽pS4-1和4型HEVORF2C-端385～674aa多肽pS4-6分别包被微孔板,采用间接酶联免疫法(EIA)检测感染HEV的猴系列血清中的抗HEVIgG抗体,分析两种多肽的抗原性,并与进口试剂的检测结果进行比较。结果在感染HEV的猴系列血清中,针对HEVORF2N-端抗原的抗体产生时间早,持续时间短,其峰值与转氨酶(ALT)的峰值几乎重叠;针对HEVORF2C-端抗原的抗体在HEV感染早期产生的滴度较低,但逐渐升高,在14周的观察期内,抗体滴度几乎无下降趋势。用两种多肽检测HEVIgG抗体的灵敏度均高于进口试剂。结论两种多肽均具有较好的抗原性,但在HEV感染的不同时期,其反应性不同。     

应用层析技术提高低pH静脉注射丙种球蛋白制剂质量

目的　进一步提高低ｐＨ静脉注射两种球蛋白制剂质量。方法采用一步ＤＥＡＥ－Ｓｅｐｈａｒｏｓｅ　ＦＦ柱 层析吸附。结果ＩｇＡ残留量由吸附前的５．６０±３．ｍｇ／ｇ蛋白下降到１．１６±０．３８ｍｇ／ｇ蛋白,清除率达（７５．３２± １９．１９）％,总蛋白回收达（８８．５５±２．７６）％,ＩｇＧ纯度进一步提高。制剂中的ＡＣＡ、ＰＫＡ、白喉抗体、单体加二聚体比例等指标均符合规程要求。结论 ＤＥＡＥ－Ｓｅｐｈａｒｏｓｅ柱层析吸附可提高ＩＶＩｇ质量。     

Neuroimmunological Implications of AQP4 in Astrocytes

The brain has high-order functions and is composed of several kinds of cells, such as neurons and glial cells. It is becoming clear that many kinds of neurodegenerative diseases are more-or-less influenced by astrocytes, which are a type of glial cell. Aquaporin-4 (AQP4), a membrane-bound protein that regulates water permeability is a member of the aquaporin family of water channel proteins that is expressed in the endfeet of astrocytes in the central nervous system (CNS). Recently, AQP4 has been shown to function, not only as a water channel protein, but also as an adhesion molecule that is involved in cell migration and neuroexcitation, synaptic plasticity, and learning/memory through mechanisms involved in long-term potentiation or long-term depression. The most extensively examined role of AQP4 is its ability to act as a neuroimmunological inducer. Previously, we showed that AQP4 plays an important role in neuroimmunological functions in injured mouse brain in concert with the proinflammatory inducer osteopontin (OPN). The aim of this review is to summarize the functional implication of AQP4, focusing especially on its neuroimmunological roles. This review is a good opportunity to compile recent knowledge and could contribute to the therapeutic treatment of autoimmune diseases through strategies targeting AQP4. Finally, the author would like to hypothesize on AQP4’s role in interaction between reactive astrocytes and reactive microglial cells, which might occur in neurodegenerative diseases. Furthermore, a therapeutic strategy for AQP4-related neurodegenerative diseases is proposed.     

Evaluation of Orbital Lymphoproliferative and Inflammatory Disorders by Gene Expression Analysis

Non-specific orbital inflammation (NSOI) and IgG4-related orbital disease (IgG4-ROD) are often challenging to differentiate. Furthermore, it is still uncertain how chronic inflammation, such as IgG4-ROD, can lead to mucosa-associated lymphoid tissue (MALT) lymphoma. Therefore, we aimed to evaluate the diagnostic value of gene expression analysis to differentiate orbital autoimmune diseases and elucidate genetic overlaps. First, we established a database of NSOI, relapsing NSOI, IgG4-ROD and MALT lymphoma patients of our orbital center (2000–2019). In a consensus process, three typical patients of the above mentioned three groups (mean age 56.4 ± 17 years) at similar locations were selected. Afterwards, RNA was isolated using the RNeasy FFPE kit (Qiagen) from archived paraffin-embedded tissues. The RNA of these 12 patients were then subjected to gene expression analysis (NanoString nCounter^®), including a total of 1364 target genes. The most significantly upregulated and downregulated genes were used for a machine learning algorithm to distinguish entities. This was possible with a high probability (p < 0.0001). Interestingly, gene expression patterns showed a characteristic overlap of lymphoma with IgG4-ROD and NSOI. In contrast, IgG4-ROD shared only altered expression of one gene regarding NSOI. To validate our potential biomarker genes, we isolated the RNA of a further 48 patients (24 NSOI, 11 IgG4-ROD, 13 lymphoma patients). Then, gene expression pattern analysis of the 35 identified target genes was performed using a custom-designed CodeSet to assess the prediction accuracy of the multi-parameter scoring algorithms. They showed high accuracy and good performance (AUC ROC: IgG4-ROD 0.81, MALT 0.82, NSOI 0.67). To conclude, genetic expression analysis has the potential for faster and more secure differentiation between NSOI and IgG4-ROD. MALT-lymphoma and IgG4-ROD showed more genetic similarities, which points towards progression to lymphoma.     

Catalytic Antibodies in Bipolar Disorder: Serum IgGs Hydrolyze Myelin Basic Protein

The pathogenesis of bipolar affective disorder is associated with immunological imbalances, a general pro-inflammatory status, neuroinflammation, and impaired white matter integrity. Myelin basic protein (MBP) is one of the major proteins in the myelin sheath of brain oligodendrocytes. For the first time, we have shown that IgGs isolated from sera of bipolar patients can effectively hydrolyze human myelin basic protein (MBP), unlike other test proteins. Several stringent criteria were applied to assign the studied activity to serum IgG. The level of MBP-hydrolyzing activity of IgG from patients with bipolar disorder was statistically significantly 1.6-folds higher than that of healthy individuals. This article presents a detailed characterization of the catalytic properties of MBP-hydrolyzing antibodies in bipolar disorder, including the substrate specificity, inhibitory analysis, pH dependence of hydrolysis, and kinetic parameters of IgG-dependent MBP hydrolysis, providing the heterogeneity of polyclonal MBP-hydrolyzing IgGs and their difference from canonical proteases. The ability of serum IgG to hydrolyze MBP in bipolar disorder may become an additional link between the processes of myelin damage and inflammation.     

Distinct Longitudinal Changes in Immunoglobulin G N-Glycosylation Associate with Therapy Response in Chronic Inflammatory Diseases

Immunosuppressants and biologicals are widely used therapeutics for various chronic inflammatory diseases (CID). To gain more detailed insight into their downstream effects, we examined their impact on serum immunoglobulin G (IgG) glycosylation. We analyzed IgG subclass-specific fragment crystallizable (Fc) N-glycosylation in patients suffering from various CID using the LC-MS approach. Firstly, we compared IgG Fc N-glycosylation between 128 CID patients and 204 healthy controls. Our results replicated previously observed CID-related decrease in IgG Fc galactosylation (adjusted p-value range 1.70 × 10⁻²–5.95 × 10⁻²²) and sialylation (adjusted p-value range 1.85 × 10⁻²–1.71 × 10⁻¹⁸). Secondly, to assess changes in IgG Fc N-glycosylation associated with therapy and remission status, we compared 139 CID patients receiving either azathioprine, infliximab, or vedolizumab therapy. We observed an increase in IgG Fc galactosylation (adjusted p-value range 1.98 × 10⁻²–1.30 × 10⁻¹⁵) and sialylation (adjusted p-value range 3.28 × 10⁻⁶–4.34 × 10⁻¹⁸) during the treatment. Furthermore, patients who reached remission displayed increased Fc galactosylation levels (p-value range 2.25 × 10⁻²–5.44 × 10⁻³) in comparison to patients with active disease. In conclusion, the alterations in IgG Fc glycosylation and the fact these changes are even more pronounced in patients who achieved remission, suggest modulation of IgG inflammatory potential associated with CID therapy.     

一种广泛适用的IgG敏感膜压电免疫传感器

压电晶振ＱＣＭ金电极表面采用金－蛋白质Ａ－ＩｇＧ的方法制备生物敏感膜,选择不同种类的ＩｇＧ,可检测相应的不同种类的抗原或抗体,适用范围广泛。文中讨论了蛋白质Ａ总量、ＩｇＧ浓度对压电免疫传感器频移的影响,采用两种方法测定了对不同浓度待测度的响应曲线和灵敏度,考察了特定的敏感膜对于相应抗原或抗体的特异性识别能力。     

Covalent immobilization of antibodies for the preparation of immunoaffinity chromatographic supports

Immunosorbents in immunoaffinity chromatography (IAC) are prepared by immobilizing expensive antibodies without guidelines for ensuring the best coupling efficiencies, and avoiding low binding capacities. Covalent immobilization of antibodies on N-hydroxysuccinimide (NHS)-activated Sepharose 4 Fast Flow resin was optimized using human IgG via full factorial design with incubation times (4, 9, 14, 19 and 24 h), temperatures (4°C and 20°C) and coupling reaction buffers (sodium bicarbonate and triethanolamine). The best coupling efficiency (CE) (83.4 ± 8.7%) was reached with triethanolamine buffer, 14 h and 4°C. Comparison of antibody isotypes (IgG or IgM) by a nested factorial analysis suggested that antibodies in the IgG isotype presents the best coupling efficiency.     

牛初乳巴氏杀菌工艺的优化研究

热处理是牛初乳加工中必不可少的工艺过程.通过对热处理强度对初乳中IgG保留率、微生物数量以及营养成分损失的影响进行研究,并对巴式杀菌工艺进行优化,得出以下结果:杀菌温度为60℃和63℃,加热时间超过30 min可以将微生物数量降低到安全数量.IgG的保留率随加热温度的升高而降低,随加热时间的延长而降低.加热温度对美拉德反应、脂肪氧化和蛋白质水解反应的影响均显著.通过优化得出,杀菌温度60℃和加热时间30 min,可以既保证微生物安全,同时最大保留IgG的浓度和营养成分损失较低.     

静脉滴注丙种球蛋白治疗急性毛细支气管炎的研究

应用静脉滴注丙种球蛋白（IVIg）治疗毛细支气管炎25例,以30例应用新鲜血浆或血液者为对照,结果无论是临床症状、体征,还是体液免疫功能变化,均较对照组好转、恢复快,并且没有出现任何后遗症和死亡病例。