Studies on the antiinflammatory,immunoregulatory, and analgesic actions of melatonin

In order to develop melatonin (MT) as a potential new drug for the treatment of diseases with inflammation, pain, and abnormal immune responses, the effects and mechanisms of MT on inflammation, immunoregulation, and nociception were studied systematically. MT (40–160 mg/kg, ip) had significant analgesic effects in the hot-plate, writhing, and tail-flick models, with a marked dose- and time-dependence. The onset of its analgesia about 30–60 min after ip, was slower than that of pethidine, but the duration was longer (about 1.5–2 h). The analgesia was also induced by icv MT (0.25 mg/kg) injection. A lower dose of MT (10 mg/kg) could enhance the analgesic effect of pethidine, which was blocked by naloxone (10 mg/kg). MT (100 mg/kg, ip) decreased the content of beta-endorphin in the hypothalamus and pituitary. The analgesia of MT could be attenuated by pretreatment with reserpine (30 mg/kg, ip) or phentolamine (10 mg/kg, ip). CaCl₂ (230 mg/kg) could antagonize the analgesia of MT. EGTA and verapamil had opposite effects to CaCl₂. No tolerance and dependence to MT was found in mice. Further studies showed that MT could enhance the functions of T and B lymphocytes and macrophages in vitro and in adjuvant arthritis, and inhibit the disturbance of immune cells. MT could inhibit the swelling of hindpaw induced by carrageenin and complete Freund's adjuvant. These factors suggest that MT possesses marked antiinflammatory, immunoregulatory, and analgesic effects which may be related to the system of opiate, monoamine, and Ca²⁺ modulation. Drug Dev. Res. 39:167–173. © 1997 Wiley-Liss, Inc.     

水凝胶负载pVAX1-SpaP/P防龋DNA疫苗以4种途径产生免疫效应

背景:防龋DNA疫苗价格低廉、安全有效,有望成为人类抵抗龋病的重要手段,因而学者在寻找一种能高效诱导抗体产生的免疫途径或保护疫苗效价的缓释系统及佐剂,成为近年研究的热点.目的:观察聚乳酸乙醇酸-聚乙二醇-聚乳酸乙醇酸水凝胶负载pVAX1-SpaP/P防龋基因疫苗的免疫效应.方法:以聚乳酸乙醇酸-聚乙二醇-聚乳酸乙醇酸水...     

Diagnostic utility of programmed cell death ligand 1 (clone SP142) immunohistochemistry for malignant lymphoma and lymphoproliferative disorders: A brief review

The programmed cell death 1 (PD1)/PD1 ligand (PD-L1) axis plays an important role in tumor cell escape from immune control and has been most extensively investigated for therapeutic purposes. However, PD-L1 immunohistochemistry is still not used widely for diagnosis. We review the diagnostic utility of PD-L1 (by clone SP142) immunohistochemistry in large-cell lymphomas, mainly consisting of classic Hodgkin lymphoma (CHL) and diffuse large B-cell lymphoma (DLBCL). Neoplastic PD-L1 (nPD-L1) expression on Hodgkin and Reed-Sternberg cells is well-established among prototypic CHL. Of note, EBV+ CHL often poses a challenge for differential diagnosis from peripheral T-cell lymphoma with EBV+ non-malignant large B-cells; their distinction is based on the lack of PD-L1 expression on large B-cells in the latter. The nPD-L1 expression further provides a good diagnostic consensus for CHL with primary extranodal disease conceivably characterized by a combined pathogenesis of immune escape of tumor cells and immunodeficiency. Compared with CHL, the nPD-L1 expression rate is much lower in DLBCL, highlighting some specific subgroups of intravascular large B-cell lymphoma, primary mediastinal large B-cell lymphoma, and EBV+ DLBCL. They consist of nPD-L1-positive and -negative subgroups, but their clinicopathological significance remains to be elucidated. Microenvironmental PD-L1 positivity on immune cells may be associated with a favorable prognosis in extranodal DLBCL. PD-L1 (by SP142) immunohistochemistry has helped us to understand the immune biology of lymphoid neoplasms possibly related by immune escape and/or immunodeficiency. However, knowledge of these issues remains limited and should be clarified for diagnostic consensus in the future.     

肝移植106例术后急性排斥反应的诊断和治疗

目的:总结肝移植急性排斥反应的新表现及诊治经验. 方法:回顾我中心106例肝移植资料,对其中发生急性排斥反应病例的临床表现、病理组织学改变、诊治方案加以分析. 结果:106例肝移植中,术后发生急性排斥反应17例(16.0%);其中临界型1例,轻度6例,中度7例,重度3例. 这些病例在具备病理组织学改变的同时缺乏典型的临床特征和肝功能生化指标的改变. 结论:由于强效免疫抑制剂的应用,肝移植术后急性排斥反应发生的临床症状和体征不典型,我们应该重视急性排斥反应的诊断并积极治疗.     

活动性肺结核患者血清炎性因子变化特点及与免疫状态的关系

目的探讨活动性结核患者血清炎性因子水平特征及与患者免疫状态的关系.方法选择2003年2月至2005年10月本科住院的结核患者97例,其中活动型结核患者57例,静止期患者40例,对照组为41例健康成年体检者.ELISA法及碱性磷酸酶抗碱性磷酸酶(APAAP)法检测活动性肺结核患者血清中炎性因子水平(TNF-α、IL-1和IL-6)及相关细胞免疫指标(CD4、CD8、CD4/CD8)的变化.结果正常对照组IL-1、IL-6、TNF-α水平分别为(15.3±1.3)、(80.5±7.3)、(77.2±9.8)ng /ml,活动性结核组为(33.7±3.6)、(293.6±30.5)和(190.7±25.2)ng/ml,显著高于正常对照组和静止性结核组.活动性结核组患者外周血CD4比例及CD4/CD8比值为(32.3±2.9)%和(0.83±0.17),显著低于正常对照组.结论炎性因子水平增高及免疫功能下降是活动型结核患者的临床特点之一,两者呈反向依赖关系.     

Porcine TRIM21 Enhances Porcine Circovirus 2 Infection and Host Immune Responses,But Inhibits Apoptosis of PCV2-Infected Cells

Tripartite motif protein 21 (TRIM21) is an interferon-inducible E3 ligase, containing one RING finger domain, one B-box motif, one coiled-coil domain at the N-terminal, as well as one PRY domain and one SPRY domain at the C-terminal. TRIM21 is expressed in many tissues and plays an important role in systemic autoimmunity. However, TRIM21 plays different roles in different virus infections. In this study, we evaluate the relationship between porcine TRIM21 and PCV2 infection as well as host immune responses. We found that PCV2 infection modulated the expression of porcine TRIM21. TRIM21 can enhance interferons and proinflammatory factors and decrease cellular apoptosis in PCV2-infected cells. These results indicate that porcine TRIM21 plays a critical role in enhancing PCV2 infection, which is a promising target for controlling and developing the treatment of PCV2 infection.     

IFI27 is a potential therapeutic target for HIV infection

BackgroundTherapeutic studies against human immunodeficiency virus type 1 (HIV-1) infection have become one of the important works in global public health.MethodsDifferential expression analysis was performed between HIV-positive (HIV+) and HIV-negative (HIV-) patients for {"type":"entrez-geo","attrs":{"text":"GPL6947","term_id":"6947"}}GPL6947 and {"type":"entrez-geo","attrs":{"text":"GPL10558","term_id":"10558"}}GPL10558 of {"type":"entrez-geo","attrs":{"text":"GSE29429","term_id":"29429"}}GSE29429. Coexpression analysis of common genes with the same direction of differential expression identified modules. Module genes were subjected to enrichment analysis, Short Time-series Expression Miner (STEM) analysis, and PPI network analysis. The top 100 most connected genes in the PPI network were screened to construct the LASSO model, and AUC values were calculated to identify the key genes. Methylation modification of key genes were identified by the chAMP package. Differences in immune cell infiltration between HIV + and HIV- patients, as well as between antiretroviral therapy (ART) and HIV + patients, were calculated using ssGSEA.ResultsWe obtained 3610 common genes, clustered into nine coexpression modules. Module genes were significantly enriched in interferon signalling, helper T-cell immunity, and HIF-1-signalling pathways. We screened out module genes with gradual changes in expression with increasing time from HIV enrolment using STEM software. We identified 12 significant genes through LASSO regression analysis, especially proteasome 20S subunit beta 8 (PSMB8) and interferon alpha inducible protein 27 (IFI27). The expression of PSMB8 and IFI27 were then detected by quantitative real-time PCR. Interestingly, IFI27 was also a persistently dysregulated gene identified by STEM. In addition, 10 of the key genes were identified to be modified by methylation. The significantly infiltrated immune cells in HIV + patients were restored after ART, and IFI27 was significantly associated with immune cells.ConclusionThe above results provided potential target genes for early diagnosis and treatment of HIV + patients. IFI27 may be associated with the progression of HIV infection and may be a powerful target for immunotherapy.     

Paraneoplastic Opsoclonus-myoclonus Syndrome with Anti-Hu and Anti-SOX-1 Antibodies after Immune-checkpoint Inhibitor Treatment Combined with Chemotherapy in a Patient with Small-cell Lung Cancer

A 69-year-old man with advanced small-cell lung cancer achieved partial remission after 3 courses of immunochemotherapy that included atezolizumab. Ten days after the last treatment, he developed paraneoplastic opsoclonus-myoclonus syndrome and required mechanical ventilation. Serology testing detected anti-Hu and anti-SOX-1 antibodies. Despite steroid pulse therapy, various anticonvulsants, continuous intravenous sedation, and a fourth course of chemotherapy without atezolizumab, his condition failed to improve. Paraneoplastic opsoclonus-myoclonus syndrome with autoantibodies after immune-checkpoint inhibitor treatment has not been reported previously. Although a causal relationship between immune-checkpoint inhibitors and paraneoplastic syndromes has been suggested, the mechanism remains unknown.     

In situ proximity labeling identifies Lewy pathology molecular interactions in the human brain

The intracellular misfolding and accumulation of alpha-synuclein into structures collectively called Lewy pathology (LP) is a central phenomenon for the pathogenesis of synucleinopathies, including Parkinson’s disease (PD) and dementia with Lewy bodies (DLB). Understanding the molecular architecture of LP is crucial for understanding synucleinopathy disease origins and progression. Here we used a technique called biotinylation by antibody recognition (BAR) to label total (BAR-SYN1) and pathological alpha-synuclein (BAR-PSER129) in situ for subsequent mass spectrometry analysis. Results showed superior immunohistochemical detection of LP following the BAR-PSER129 protocol, particularly for fibers and punctate pathology within the striatum and cortex. Mass spectrometry analysis of BAR-PSER129–labeled LP identified 261 significantly enriched proteins in the synucleinopathy brain when compared to nonsynucleinopathy brains. In contrast, BAR-SYN1 did not differentiate between disease and nonsynucleinopathy brains. Pathway analysis of BAR-PSER129–enriched proteins revealed enrichment for 718 pathways; notably, the most significant KEGG pathway was PD, and Gene Ontology (GO) cellular compartments were the vesicle, extracellular vesicle, extracellular exosome, and extracellular organelle. Pathway clustering revealed several superpathways, including metabolism, mitochondria, lysosome, and intracellular vesicle transport. Validation of the BAR-PSER129–identified protein hemoglobin beta (HBB) by immunohistochemistry confirmed the interaction of HBB with PSER129 Lewy neurites and Lewy bodies. In summary, BAR can be used to enrich for LP from formalin-fixed human primary tissues, which allowed the determination of molecular signatures of LP. This technique has broad potential to help understand the phenomenon of LP in primary human tissue and animal models.

Parkinson’s disease (PD) and dementia with Lewy bodies (DLB) are progressive neurodegenerative diseases characterized by intracellular inclusions containing aggregated alpha-synuclein referred to as Lewy pathology (LP). Although strong evidence suggests alpha-synuclein is involved in the disease process, central questions remain regarding the molecular origins and progression of LP. A diverse range of cellular pathways have been implicated in alpha-synuclein–mediated disease processes including dysfunctional synaptic neurotransmission and vesicular trafficking, mitochondrial dysfunction, oxidative stress, neuroinflammation, calcium signaling, glycolysis, autophagy lysosomal dysfunction, and Mitogen-activated protein kinase (MAPK) signaling pathways (1). However, much of the evidence implicating these cellular processes has been derived from cell and animal models, which may or may not closely recapitulate the human disease processes. Defining the molecular details of disease process is important for progress toward the development of disease-modifying therapies.Alpha-synuclein phosphorylated at serine 129 (PSER129) is a marker of LP. PSER129 is enriched in LP with only trace quantities (<5% of total alpha-synuclein pool) of endogenous alpha-synuclein being phosphorylated (2). Evidence suggests that PSER129 may aggregate more rapidly, and PSER129 may have a role in regulating alpha-synuclein turnover (3). Evidence also suggests that PSER129 accumulates in the brain as PD progresses (4, 5) although it is not clear whether PSER129 occurs prior to, during, or following inclusion formation (6). Several antibodies have been developed to detect PSER129, including the commonly used EP1536Y antibody (Abcam), a rabbit monoclonal antibody against PSER129 that has high specificity for LP (7) and has been used extensively for immunohistochemical (IHC) labeling of LP.Determining the molecular makeup of LP in the human brain is challenging. Difficulties arise principally from the insoluble nature of LP, which precludes common biochemical methods for determining molecular interactions. In past studies, LP from the human brain has been enriched for by biochemical fractionation methods (8) and isolated by microdissection techniques (9). There are important limitations to each approach. Fractionation disturbs cellular structures and, presumably, LP composition. Microdissection exclusively captures Lewy bodies, but not other lesions (neurites, dots, etc.). Microscopy methods provide accurate data characterizing spatial and molecular makeup of LP, but these methods are limited to known molecular targets. Together, current approaches are insufficient to identify molecular details of LP in the human diseased brain.Recently, a technique referred to as biotinylation by antibody recognition (BAR) was successfully used to label insoluble cellular components within proximity to an antigen in fixed primary tissues for enrichment and downstream mass spectrometry analysis (10–13). The BAR technique (also called selective proteomic proximity labeling using tyramide, or enzyme-mediated activation of radical sources) labels a target antigen by peroxidase-catalyzed deposition of biotin moieties onto proximal molecules. BAR is particularly well suited for determining protein interactions and subcellular localization of insoluble cellular components (10). LPs are insoluble cellular inclusions, and therefore the BAR technique holds promise to study these structures. Recent demonstrations of the BAR technique, and its utility for proteomic analysis, gave us the impetus to attempt to purify LP and LP interactors from fixed primary human tissues.Here we applied a modified BAR technique to capture total alpha-synuclein (BAR-SYN1) and LP (BAR-PSER129) interactions in formalin-fixed human brain samples from individuals diagnosed with PD and DLB (SYN), as well as neurologically intact individuals (HC).