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Caroline Rouard, Nadège Bourgeois-Nicolaos, Lalaina Rahajamanana, Olivier Romain, Lydia Pouga, Véronique Derouin, Daniele De Luca, Florence Doucet-Populaire
BACKGROUND: Meticillin-susceptible and resistant Staphylococcus aureus (MSSA and MRSA) are responsible for outbreaks in intensive care units. MSSA infections have the same morbidity and mortality rate as MRSA infections but are less often studied. Whole genome sequencing (WGS) is increasingly used for outbreak monitoring but still requires specific installation and trained personnel to obtain and analyse the data. AIM: To evaluate the workflow and the benefits of the bioMerieux Episeq® (bioMerieux, France) solution in exploring an increased incidence of S...
February 13, 2019: Journal of Hospital Infection
Yuta Yamamoto, Tetsuya Saita, Rintaro Sogawa, Kenji Ogata, Yutaro Yamamoto, Sakiko Kimura, Yutaka Narisawa, Shinya Kimura, Masashi Shin
The tyrosine kinase inhibitor ponatinib is extensively metabolized in the body, and consequently the development of specific immunoassays for pharmacokinetic studies and therapeutic drug monitoring of ponatinib is challenging. If two antibodies simultaneously recognize the entire structure of ponatinib, they could be utilized to establish an ultra-specific sandwich immunoassay for ponatinib, free of any interference from ponatinib metabolites. In this study, we created two types of anti-ponatinib polyclonal antibodies that recognize two different ponatinib epitopes, and sandwiched almost all structural components of ponatinib in these two antibodies in order to develop an enzyme-linked immunosorbent assay (ELISA) technique not affected by any ponatinib metabolites...
February 13, 2019: Analytical Biochemistry
Zhenwu Luo, Min Li, Yongxia Wu, Zhefeng Meng, Lisa Martin, Lumin Zhang, Elizabeth Ogunrinde, Zejun Zhou, Shenghui Qin, Zhuang Wan, Maria Anna Julia Westerink, Stephanie Warth, Hui Liu, Ping Jin, David Stroncek, Quan-Zhen Li, Ena Wang, Xueling Wu, Sonya L Heath, Zihai Li, Alexander V Alekseyenko, Wei Jiang
BACKGROUND: Increased autoreactive antibodies have been reported in HIV disease; however, the mechanism accounting for autoantibody induction in HIV remains unknown. RESULTS: Herein, we show that seasonal influenza vaccination induces autoantibody production (e.g., IgG anti-nuclear antibody (ANA) and anti-double-stranded DNA antibody (anti-dsDNA)) in some viral-suppressed antiretroviral therapy (ART)-treated HIV+ subjects, but not in healthy controls. These autoantibodies were not derived from antigen-specific B cells but from activated "bystander" B cells analyzed by single-cell assay and by study of purified polyclonal ANAs from plasma...
February 14, 2019: Microbiome
Jon Von Visger, Clarissa Cassol, Uday Nori, Gerardo Franco-Ahumada, Tibor Nadasdy, Anjali A Satoskar
BACKGROUND: Proliferative glomerulonephritis with monoclonal IgG deposits (PGNMIGD) is a disease entity classified under the group of "Monoclonal gammopathy-related kidney diseases", and can recur after transplant. Clinical remission of proteinuria in patients with PGNMIGD has been previously shown following anti-B cell and/or anti-plasma cell therapies. Our case is the first to show complete histologic resolution of the glomerular monoclonal IgG kappa deposits in a case of recurrent PGNMIGD in renal allograft after rituximab and steroid treatment...
February 14, 2019: BMC Nephrology
Antonia Terzieva, Violeta Dimitrova, Lyubomir Djerov, Petya Dimitrova, Silvina Zapryanova, Iana Hristova, Ivaylo Vangelov, Tanya Dimova
Pregnancy is a state where high and stage-dependent plasticity of the maternal immune system is necessary in order to equilibrate between immunosuppression of harmful responses towards the fetus and ability to fight infections. TCR γδ cells have been implicated in the responses in infectious diseases, in the regulation of immune responses, and in tissue homeostasis and repair. The variety of functions makes γδ T cells a particularly interesting population during pregnancy. In this study, we investigated the proportion, phenotype and TCR γ and δ repertoires of γδ T cells at the maternal⁻fetal interface and in the blood of pregnant women using FACS, immunohistochemistry and spectratyping...
February 5, 2019: International Journal of Molecular Sciences
Mohamed H Shamji, Irene Thomsen, Janice Layhadi, Jasper Kappen, Gabriële Holtappels, Umit Sahiner, Amy Switzer, Stephen R Durham, Oliver Pabst, Claus Bachert
BACKGROUND: Chronic rhinosinusitis with nasal polyps (CRSwNP) is often characterized by local production of polyclonal IgE-idiotypes. Whilst tissue IgE concentrations can be in the range of several thousand kU/L, the regulatory mechanisms by which IgE-mediated inflammation is controlled in the nasal polyps is not well understood. OBJECTIVE: We sought to determine whether locally induced IgG antibodies in the nasal polyps can inhibit IgE-mediated pro-allergic response...
February 11, 2019: Journal of Allergy and Clinical Immunology
Mark Angelo V Ngu, Jose H Bergantin, John Donnie A Ramos
The house dust mite Suidasia pontifica (Sp) is an important source of allergens in tropical regions that trigger IgE-mediated allergic reactions such as allergic asthma, atopic dermatitis and allergic rhinitis. Detection of Sp-specific proteins are important in the management and prevention of allergic diseases. The study aimed to provide a proof of concept for a gold nanoparticle-labeled sandwich format lateral flow immunoassay (LFIA) kit for the detection of Sp-specific proteins. Protein A chromatography-purified rabbit anti-Sp polyclonal antibodies were labeled with gold nanoparticles (AuNP) synthesized from chloroauric acid using the citrate reduction method, then dispensed on a glass fiber pad...
February 12, 2019: Protein and Peptide Letters
Yu Wen, He N Xu, Lisa Privette Vinnedge, Min Feng, Lin Z Li
PURPOSE: Optical redox imaging (ORI), based on collecting the endogenous fluorescence of reduced nicotinamide adenine dinucleotide (NADH) and oxidized flavoproteins (Fp) containing a redox cofactor flavin adenine dinucleotide (FAD), provides sensitive indicators of cellular metabolism and redox status. ORI indices (such as NADH, FAD, and their ratio) have been under investigation as potential progression/prognosis biomarkers for cancer. Higher FAD redox ratio (i.e., FAD/(FAD + NADH)) has been associated with higher invasive/metastatic potential in tumor xenografts and cultured cells...
February 13, 2019: Molecular Imaging and Biology: MIB: the Official Publication of the Academy of Molecular Imaging
A M Heeren, J Rotman, A G M Stam, N Pocorni, A A Gassama, S Samuels, M C G Bleeker, C H Mom, H J M A A Zijlmans, G G Kenter, E S Jordanova, T D de Gruijl
BACKGROUND: Cervical cancer (CxCa) is mainly a locally invading disease that metastasizes to loco-regional lymph node basins before involving distant organs in more advanced stages. Local immune potentiation of tumor-draining lymph nodes (TDLN) may thus protect against tumor progression. METHODS: To identify therapeutic targets for local immune modulation, multi-parameter flow cytometric T-cell profiling of primary cervical tumors (PT) and TDLN (n = 37) was performed...
February 12, 2019: Journal for Immunotherapy of Cancer
Kulachelvy Ratnasothy, Jacintha Jacob, Sim Tung, Dominic Boardman, Robert Ian Lechler, Alberto Sanchez Fueyo, Marc Martinez-Llordella, Giovanna Lombardi
Regulatory T cells (Tregs) have unique immunosuppressive properties and are essential to ensure effective immunoregulation. In animal models, Tregs have been shown to prevent autoimmune disorders and establish transplantation tolerance. Therefore, the prospect of harnessing Tregs, either by increasing their frequency or by conferring allospecificity, has prompted a growing interest in the development of immunotherapies. Here, employing a well-established skin transplant model with a single MHC mismatch, we compared the therapeutic efficacy of adoptively transfer Treg with or without donor-specificity and the administration of IL-2 to promote in vivo expansion of Treg...
February 12, 2019: American Journal of Transplantation
Raquel Santos-de-Souza, Franklin Souza-Silva, Barbara Cristina de Albuquerque-Melo, Michelle Lopes Ribeiro-Guimarães, Luzia Monteiro de Castro Côrtes, Bernardo Acácio Santini Pereira, Mariana Silva-Almeida, Léa Cysne-Finkelstein, Francisco Odencio Rodrigues de Oliveira Junior, Mirian Claudia de Souza Pereira, Carlos Roberto Alves
Leishmania (Leishmania) amazonensis has adaptive mechanisms to the host environment that are guided by its proteinases, including cysteine proteinase B (CPB), and primarily its COOH-terminal region (Cyspep). This work aimed to track the fate of Cyspep by surface plasmon resonance (SPR) of promastigotes and amastigotes to gain a greater understanding of the adaptation of this parasite in both hosts. This strategy consisted of antibody immobilization on a COOH1 surface, followed by interaction with parasite proteins and epoxysuccinyl-L-leucylamido(4-guanidino)butane (E-64)...
February 12, 2019: Parasitology Research
Manuel Alvarez-Rodriguez, Manel López-Bejar, Heriberto Rodriguez-Martinez
Presence of the hyaluronan (hyaluronic acid, HA) receptor CD44 on spermatozoa has been difficult to pursue, mostly obeying to the use of different commercial mono- and/or polyclonal antibodies, often lacking proper controls. Here, we describe how the presence (Western blotting) and specific location (immunocytochemistry) of the CD44 receptor differs in ejaculated pig spermatozoa depending on the type of antibody and protocol used. While we were able to detect binding to spermatozoa and mark its presence in the sperm membrane, the use of blocking peptides clearly indicated that only the monoclonal antibody could confirm the specific presence and location of the CD44 receptor, whereas the polyclonal antibody was detecting multiple presumed CD44 isoforms or degraded proteins thus proving unspecific...
February 11, 2019: Reproduction in Domestic Animals, Zuchthygiene
Fleur S van de Bovenkamp, Ninotska I L Derksen, Pleuni Ooijevaar-de Heer, Theo Rispens
About 15% of immunoglobulin G (IgG) molecules contain glycans linked to the antigen-binding fragments (Fab arms) in addition to the glycans linked to the crystallizable fragment (Fc tail) of all IgGs. Fab glycosylation appears to be an important feature of antibodies, for example by influencing antigen binding and antibody stability. The reliable generation of antibodies that either have or lack Fab glycans would be very helpful to study the role of Fab glycans in more detail. In this study, we set out to remove Fab glycans by treating polyclonal and monoclonal human IgG antibodies with two commonly used glycosidases and an improved version of one of the two (Endo F3, PNGase F, and Rapid™ PNGase F)...
February 8, 2019: Journal of Immunological Methods
Mona M Al-Shehri, Adel S El-Azab, Manal A El-Gendy, Mohammed A Hamidaddin, Ibrahim A Darwish
Crizotinib (CZT) is a potent drug used for treatment of non-small cell lung cancer (NSCLC); however, its circulating concentration variability has been associated with acquired resistance and toxicity, restricting the success of cancer treatment. As such, the development of an assay that monitors CZT plasma concentrations in patients is a valuable tool in cancer treatment. In this study, a hapten of CZT was synthesized by introducing the acetohydrazide moiety as a spacer into the chemical structure of CZT. The chemical structure of the CZT acetohydrazide (hapten) was confirmed by mass, 1H-, and 13C-NMR spectrometric techniques...
2019: PloS One
Olga Taraschenko, Howard S Fox, Sean J Pittock, Anastasia Zekeridou, Maftuna Gafurova, Ember Eldridge, Jinxu Liu, Shashank M Dravid, Raymond Dingledine
OBJECTIVE: Seizures develop in 80% of patients with anti-N-methyl-d-aspartate receptor (NMDAR) encephalitis, and these represent a major cause of morbidity and mortality. Anti-NMDAR antibodies have been linked to memory loss in encephalitis; however, their role in seizures has not been established. We determined whether anti-NMDAR antibodies from autoimmune encephalitis patients are pathogenic for seizures. METHODS: We performed continuous intracerebroventricular infusion of cerebrospinal fluid (CSF) or purified immunoglobulin (IgG) from the CSF of patients with anti-NMDAR encephalitis or polyclonal rabbit anti-NMDAR IgG, in male C57BL/6 mice...
February 11, 2019: Epilepsia
Shan Li, Pengtao Gong, Nan Zhang, Xin Li, Lixin Tai, Xu Wang, Zhengtao Yang, Ju Yang, Xingquan Zhu, Xichen Zhang, Jianhua Li
Neospora caninum is an obligate intracellular apicomplexan parasite, the etiologic agent of neosporosis, and a major cause of reproductive loss in cattle. There is still a lack of effective prevention and treatment measures. The 14-3-3 protein is a widely expressed acidic protein that spontaneously forms dimers within apicomplexan parasites. This protein has been isolated and sequenced in many parasites; however, there are few reports about the N. caninum 14-3-3 protein. Here, we successfully expressed and purified a recombinant fusion protein of Nc14-3-3 (rNc14-3-3) and prepared a polyclonal antibody...
2019: Frontiers in Microbiology
S Ekman, A Lindahl, U Rüetschi, A Jansson, K Björkman, K Abrahamsson-Aurell, S Björnsdóttir, M Löfgren, L Mattsson Hultén, E Skiöldebrand
BACKGROUND: Molecular serum markers that can identify early reversible osteoarthritis (OA) in horses are lacking. OBJECTIVES: We studied serum concentrations of a novel cartilage oligomeric matrix protein (COMP) neo-epitope in horses subjected to short-term exercise and with acute lameness. The effects of circadian rhythm and age were also evaluated. STUDY DESIGN: Longitudinal studies in healthy horses and cross-sectional comparison of lame and non-lame horses...
February 10, 2019: Equine Veterinary Journal
Bin Yan, Yang He, Shi-Qi Lu, Meng-Qiao Xu, Qi Wang, Yi-Ming Zhao, Chang-Geng Ruan
OBJECTIVE: To establish a novel flow cytometric immunobead array (FCIA) for detecting plasma von Willebrand factor activity (vWF:GPIbR) and apply it in ischemic stroke (IS). METHODS: Microspheres coated with anti-human platelet glycoprotein Ibα (GPIbα) monoclonal antibody SZ151 IgG, were incubated with recombinant fragment of GPIbα, then added ristocetin and plasma, finally incubated with FITC-conjugated sheep-anti-human vWF IgG polyclonal antibody, and detected by flow cytometry...
February 2019: Zhongguo Shi Yan Xue Ye Xue za Zhi
Stefanie Graeter, Hans-Uwe Simon, Stephan von Gunten
Granulocytes are key effector cells of the innate immune system that cause substantial by-stander tissue damage in a broad range of inflammatory disorders. Specific antibodies with the potential to regulate granulocyte survival are present in polyclonal intravenous immunoglobulin (IVIG) preparations and may contribute to the anti-inflammatory effects of high-dose IVIG therapy. Here, we discuss idiosyncratic characteristics of antibodies to FAS and Siglecs contained in IVIG pertaining to granulocyte death regulation and highlight implications for research and clinical practice...
February 6, 2019: Pharmacological Research: the Official Journal of the Italian Pharmacological Society
Seçkin Akgül, Ann-Marie Patch, Rochelle C J D'Souza, Pamela Mukhopadhyay, Katia Nones, Sarah Kempe, Stephen H Kazakoff, Rosalind L Jeffree, Brett W Stringer, John V Pearson, Nicola Waddell, Bryan W Day
Glioblastomas are the most common and lethal neoplasms of the central nervous system. Neighbouring glioma cells maintain extreme degrees of genetic and phenotypic variation that form intratumoural heterogeneity. This genetic diversity allows the most adaptive tumour clones to develop treatment resistance, ultimately leading to disease recurrence. We aimed to model this phenomenon and test the effectiveness of several targeted therapeutic interventions to overcome therapy resistance. Heterogeneous tumour masses were first deconstructed into single tumour cells, which were expanded independently as single-cell clones...
February 6, 2019: Cancers
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