Mapping the tumour human leukocyte antigen (HLA) ligandome by mass spectrometry

The entirety of human leukocyte antigen (HLA)‐presented peptides is referred to as the HLA ligandome of a cell or tissue, in tumours often termed immunopeptidome. Mapping the tumour immunopeptidome by mass spectrometry (MS) comprehensively views the pathophysiologically relevant antigenic signature of human malignancies. MS is an unbiased approach stringently filtering the candidates to be tested as opposed to epitope prediction algorithms. In the setting of peptide‐specific immunotherapies, MS‐based strategies significantly diminish the risk of lacking clinical benefit, as they yield highly enriched amounts of truly presented peptides. Early immunopeptidomic efforts were severely limited by technical sensitivity and manual spectra interpretation. The technological progress with development of orbitrap mass analysers and enhanced chromatographic performance led to vast improvements in mass accuracy, sensitivity, resolution, and speed. Concomitantly, bioinformatic tools were developed to process MS data, integrate sequencing results, and deconvolute multi‐allelic datasets. This enabled the immense advancement of tumour immunopeptidomics. Studying the HLA‐presented peptide repertoire bears high potential for both answering basic scientific questions and translational application. Mapping the tumour HLA ligandome has started to significantly contribute to target identification for the design of peptide‐specific cancer immunotherapies in clinical trials and compassionate need treatments. In contrast to prediction algorithms, rare HLA allotypes and HLA class II can be adequately addressed when choosing MS‐guided target identification platforms. Herein, we review the identification of tumour HLA ligands focusing on sources, methods, bioinformatic data analysis, translational application, and provide an outlook on future developments.     

Distribution of radiolabelled anti-CA125 monoclonal antibody OC125-F(ab′)2-fragment following resection guided by antibodies (REGAJ) in ovarian cancer patients

Ovarian cancer is a highly malignant tumor of mainly postmenopausal women. The long-term prognosis of this malignancy is largely determined by micrometastasis present at the time of second-look surgery. In general, patients face a poor outcome. New radio-immunoscintigraphic methods to target tumor tissue specifically via antigen-antibody binding were developed. However, few studies so far investigated the pattern of in vivo distribution of radiolabelled mAbs and/or the specificity of antigen-antibody interaction. In this study we examined the immunological interaction and distribution of ¹³¹ I-OC125-F(ab′)₂-fragment, an anti-CA-125 mAb, in patients with CA-125 positive ovarian malignancies. Sixteen patients with primarily CA-125 positive gynecological tumors underwent REGAJ surgery. Biopsies of tumor tissue and not tumor infiltrated tissue, serum, and ascites were sampled during or prior to REGAJ surgery, respectively. After preparation of tissue cytosols, samples were assessed for CA-125 and radioactive uptake. By radiochromatography immunological analysis for presence of the target antigen CA-125, the mAb ¹³¹I-OC125-F(9ab′)2-fragment, and immune complexes was performed on different specimen. CA-125 concentrations were higher in serum samples, ascites, and malignant tissue biopsies of malignoma patients compared to those without signs of malignant disease. CA-125 was higher in the tissue cytosol than in the cell membrane fraction. Gel filtration revealed CA-125 with moieties of 75,000 to < 600,000 d. Accumulation of radioactivity was more frequently associated with the presence of unbound ¹³¹I-OC125-F(ab′)2-fragment or high molecular weight immune complexes. Radioactive uptake, however, was not confined to tissue of high CA-125 expression. Moreover, both immune complex as well as ¹³¹I-OC125-F(ab′)2-fragment could be isolated from cytosols of tissue not infiltrated by tumor cells as well. Our study demonstrates that the majority of CA-125 is located intracellularly and thus inaccessible to ¹³¹I-OC125-F(ab′)2-fragment per se. The uptake of ¹³¹I-OC125-F(ab′)2-fragment into the cytosol of tumor-free and malignant tissue samples prompts us to speculate that certain mechanisms for antigen-specific and nonspecific cellular trafficking of mAbs do exist. We present a model to explain our observations. J. Clin. Lab. Anal. 11:94–103. © 1997 Wiley-Liss, Inc.     

蒙药地格达-4味汤对TAA所致小鼠化学性肝损伤的实验研究

目的：观察蒙药地格达-4味汤对硫代乙酰胺（TAA）所致小鼠肝损伤的防治作用。方法：将地格达-4味汤组和护肝片组分别灌胃7天,腹腔注射TAA80mg.kg-1引起小鼠急性肝损害,24h后取血,分离血清,检测血清ALT、AST活性;取肝组织,观察肝脏形态学的变化。结果：地格达-4味汤组可明显降低因TAA所致肝损伤小鼠的ALT及AST（P〈0.01）。模型组肝组织学改变主要表现为碎片状坏死;护肝片组可见散在点状及小碎片状坏死;地格达-4味汤组可见散在点状坏死,有炎细胞浸润。结论：地格达-4味汤对硫代乙酰胺（TAA）所致肝损伤模型,有一定的防治作用。     

诸葛菜种子水提物对硫代乙酰胺诱导小鼠急性肝损伤的保护作用研究

该实验研究了诸葛菜种子水提物对硫代乙酰胺(TAA)诱导小鼠急性肝损伤的保护作用。将雄性ICR小鼠随机分为7组:正常组,模型组,双环醇阳性对照组(200 mg·kg-1)及葵花护肝片阳性对照组(350 mg·kg-1),诸葛菜种子水提物低(125 mg·kg-1)、中(250 mg·kg-1)、高(500 mg·kg-1)剂量组。各组按照0.02 mL·g-1灌胃给药,每日1次,连续给药4 d,第4天给药后1 h,除正常组外均腹腔注射硫代乙酰胺溶液100 mg·kg-1建立急性肝损伤模型,24 h后脱臼处死取血、取材。计算肝脏指数;检测血清中ALT,AST,TBiL活性及肝组织中SOD,GSH-Px的活性,MDA,GSH的含量;HE染色法观察肝组织病理变化;Western blot法检测NF-κB p65,Keap-1,Nrf2,p-p38,p-ERK,p-JNK,Bax,Bcl-2,caspase-3,cleaved caspase-3,caspase-8蛋白的表达。结果表明,与模型组相比,诸葛菜各给药组均能明显改善肝脏病理变化,降低肝损伤小鼠血清中ALT,AST,TBiL活性,其高剂量组肝脏组织匀浆上清中SOD与GSH-Px活性升高,GSH含量升高,MDA含量降低,同时可致肝组织中Bax,Keap-1,p-p38,p-JNK,p-ERK,NF-κB p65,cleaved caspase-3蛋白表达水平减少和Nrf2,Bcl-2,caspase-3,caspase-8蛋白表达水平增加。因此,诸葛菜种子水提物对硫代乙酰胺溶液所致小鼠急性肝损伤有明显的保护作用,其机制可能与下调Keap-1,上调Nrf2蛋白表达减轻氧化应激,抑制p38,ERK的磷酸化水平,进而抑制NF-κB信号通路的激活,抑制JNK蛋白的磷酸化,下调Bax,上调Bcl-2蛋白的表达,抑制肝细胞凋亡有关。     

Sterically stable liposomes improve the therapeutic effect of hepatic stimulator substance on fulminant hepatic failure in rats

Background and aims

Few drugs have been confirmed to be effective for fulminant hepatic failure (FHF). The purpose of this study was to prepare sterically stable liposomes (SSL) encapsulating hepatic stimulator substance (HSS) and determine their therapeutic effect on FHF.

Methods

HSS were encapsulated into SSL (HSS-SSL). FHF was induced in rats by thioacetamide (TAA) injection (400 mg/kg, three times with a 24-h interval). The agents, including HSS-SSL, SSL, HSS, and sodium chloride (NS), were each injected intravenously 2 h after the second and the third TAA injection.

Results

Freshly prepared HSS-SSL had a mean size of 93.59 nm and the average encapsulation efficiency was 37.20%. HSS encapsulated in SSL showed a longer half life and more potent target to injured livers than free HSS. Twenty-four hours after the third TAA-injection, the survival rate of HSS-SSL-treated rats (80%) was significantly higher than that of rats treated with NS (20%), SSL (25%), or HSS (50%). Histopathologic examination showed that there was the least necrosis and inflammation in the livers of HSS-SSL-treated rats. The incidence of stage 3 or 4 hepatic encephalopathy in HSS-SSL-treated rats was significantly lower than that in rats treated with other agents. The serum pro-inflammatory cytokine levels and hepatic lipid peroxidation levels were both markedly reduced, while hepatocyte proliferative rate was markedly increased after HSS-SSL treatment.

Conclusion

Encapsulation by SSL markedly improved the therapeutic effect of HSS on FHF in rats. Encapsulation by SSL may be an effective approach to enhance the therapeutic potency of drugs for FHF.     

An herbal fruit, Amomum xanthoides, ameliorates thioacetamide-induced hepatic fibrosis in rat via antioxidative system

Aim of the study

Amomum xanthoides is a well-known traditional herbal medicine mainly for diverse digestive system disorders in Asia for a long time. In the present study, we investigate the effects and action mechanism of methanol fraction of Amomum xanthoides (MFAX) on thioacetamide (TAA)-induced liver fibrosis in rat model.

Materials and methods

TAA (200 mg/kg, ip on twice a week for 14 weeks) treated rats were orally administered with MFAX (25, 50 or 100 mg/kg) once a day from the 7th week until 14th week.

Result

Significantly elevated serum bilirubin, liver tissue hydroxyproline and malondialdehyde (MDA) in liver fibrosis were ameliorated by MFAX treatment. Further, MFAX treatment attenuated the reactive oxygen species (ROS) levels and restored glutathione (GSH) content and glutathione-peroxidase (GPx) activity. Histopathological data showed that MFAX treatment inhibited collagen accumulation and activation of hepatocyte stellate cells (HSCs) in the liver tissue. Compared to the TAA group, activation of inducible nitric oxide synthase (iNOS), tumor necrosis factor alpha (TNF-α), transforming growth factor beta (TGF-β), platelet-derived growth factor beta (PDGF-β) mRNAs and the level of pro-fibrotic cytokines PDGF-β and connective tissue growth factor (CTGF) in the liver tissue were attenuated in MFAX treated groups.

Conclusion

The above evidences collectively indicate that MFAX is a potential herb which can be used as an anti-hepatofibrotic remedy.     

平肝解毒散对急性肝损伤保护作用的研究

目的：观察平肝解毒散（其主要成分柴胡）对急性肝损伤的保护作用,为临床应用提供科学依据。方法：四氯化碳（CCL4）和硫代乙酰胺（TAA）小鼠腹腔注射造成急性肝损伤实验模型,检测血清丙氨酸氨基转移酶（ALT）和天门冬氨酸氨基转移酶（AST）,并做肝脏病理切片分析。结果：CCL4模型组：平肝解毒散中、大剂量组（4g／kg、8g／kg）小白鼠的丙氨酸氨基转移酶（ALT）与模型组比较有显著差异,呈剂量依赖关系,大剂量效果最好;肝的病理切片与模型组比较肝细胞坏死程度减轻,肝细胞变性减轻。TAA模型组：平肝解毒散小剂量组小白鼠的丙氨酸氨基转移酶（ALT）和天门冬氨酸氨基转移酶（AST）与模型组比较有显著差异,小剂量有效;肝的病理切片与模型组比较肝细胞坏死程度减轻,肝细胞变性减轻。结论：平肝解毒散对CCL和TAA引起的急性肝损伤具有保护作用。     

Positive and negative factors for the treatment outcomes following total ankle arthroplasty? A systematic review

BackgroundPatient selection, surgeon’s experience and implant design play an integral role and affect the treatment outcomes of total ankle arthroplasty (TAA). The aims of this study were to investigate the positive and negative attributes that correlate with different clinical and radiographic outcomes.MethodsEight-nine studies matched the inclusion criteria: (1) studies of primary TAA with uncemented prosthesis; (2) mean follow-up of no less than 2-year; (3) reports of clinical and radiographic outcomes, and exclusion criteria: (1) non-English study; (2) more than one type of prosthesis without separated data; (3) kin studies with shorter follow-up or smaller cohort. Age, etiology, preoperative deformity, surgeon’s experience, follow-up duration and prosthetic type were studied with respect to different outcomes by mixed-effects logistic regression analysis.ResultsPatients factor: older patients reported less pain or stiffness and demonstrated less radiographic loosening which did not require additional surgical intervention. More traumatic arthritis experienced adjacent joints degeneration after TAA. Surgeon factor: less experienced surgeons had more intraoperative complications. Lack of experience for complications management without implant retrieval during early period might result in more revisions or fusion was done. Prosthetic factor: updated instrumentation decreased malalignment. If the polyethylene (PE) insert was significantly narrower than the metal components more implant instability and subsequent severe particulate wear was seen. Designs with flat-on-flat articulation and ridge at the center of the talar component associated with more PE fracture. Minimal bone resection reduced postoperative fractures. A flat cut of the tibial component and a flat undersurface with press-fit by two screws or pegs of the talar component demonstrated less postoperative fractures, whereas a syndesmosis fusion and a small triangular shape with one central fin of the talar component experienced more loosening which did not require additional surgery. Anatomic conical shape of the talar component seemed to reduce adjacent joint degeneration. Finally, fewer failures were found in patients who received HINTEGRA and Salto Talaris.ConclusionsBased on our investigation, some positive and negative factors for different clinical and radiographic outcomes were found, which should be taken into consideration in clinical practice and ankle implant design.