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Journal of Immunological Methods

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https://www.readbyqxmd.com/read/28522403/multiplex-autoantibody-detection-for-autoimmune-liver-diseases-and-autoimmune-gastritis
#1
Joris Vanderlocht, Mart van der Cruys, Frans Stals, Liesbeth Bakker-Jonges, Jan Damoiseaux
Autoantibody detection for autoimmune hepatitis (AIH), primary biliary cirrhosis (PBC) and autoimmune gastritis (AIG) is traditionally performed by IIF on a combination of tissues. Multiplex line/dot blots (LIA/DIA) offer multiple advantages, i.e. automation, objective reading, no interfering reactivities, no coincidental findings. In the current study we evaluated automated DIA (D-Tek) for detecting autoantibodies related to autoimmune diseases of the gastrointestinal tract. We tested samples of the Dutch EQC program and compared the results with the consensus of the participating labs...
May 15, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28514646/heparin-removal-by-ecteola-cellulose-pre-treatment-enables-the-use-of-plasma-samples-for-accurate-measurement-of-anti-yellow-fever-virus-neutralizing-antibodies
#2
Ana Carolina Campi-Azevedo, Vanessa Peruhype-Magalhães, Jordana Grazziela Coelho-Dos-Reis, Christiane Costa-Pereira, Anna Yoshida Yamamura, Sheila Maria Barbosa de Lima, Marisol Simões, Fernanda Freire Campos Nunes, Aline de Castro Zacche Tonini, Elenice Moreira Lemos, Ricardo Cristiano Brum, Tatiana Guimarães de Noronha, Marcos da Silva Freire, Maria de Lourdes Sousa Maia, Luiz Antônio Bastos Camacho, Maria Rios, Caren Chancey, Alessandro Romano, Carla Magda Domingues, Andréa Teixeira-Carvalho, Olindo Assis Martins-Filho
Technological innovations in vaccinology have recently contributed to bring about novel insights for the vaccine-induced immune response. While the current protocols that use peripheral blood samples may provide abundant data, a range of distinct components of whole blood samples are required and the different anticoagulant systems employed may impair some properties of the biological sample and interfere with functional assays. Although the interference of heparin in functional assays for viral neutralizing antibodies such as the functional plaque-reduction neutralization test (PRNT), considered the gold-standard method to assess and monitor the protective immunity induced by the Yellow fever virus (YFV) vaccine, has been well characterized, the development of pre-analytical treatments is still required for the establishment of optimized protocols...
May 14, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28506821/development-of-a-bioassay-as-a-measure-of-drozitumab-mediated-apoptosis-induced-by-soluble-fc-gamma-receptors
#3
Jeongsup Shim, Ally Huang, Aaron S Miller
Drozitumab is an agonistic therapeutic monoclonal antibody (mAb) against the pro-apoptotic death receptor 5 (DR5). In vitro cell killing assays using drozitumab have traditionally required cross-linking with anti-Fc antibody to amplify the pro-apoptotic signal, although drozitumab shows activity in in vivo tumor models without artificial cross-linking. Recently it has been shown that FcγR expressing cells play an important role in the activity of drozitumab by mediating cross-linking in vivo (Wilson et al...
May 12, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28502720/insights-into-the-chicken-igy-with-emphasis-on-the-generation-and-applications-of-chicken-recombinant-monoclonal-antibodies
#4
REVIEW
Warren Lee, Syed Atif Ali, Tan Soo Choon, Leow Chiuan Herng
The advantages of chicken (Gallus gallus domesticus) antibodies as immunodiagnostic and immunotherapeutic biomolecules has only been recently recognized. Even so, chicken antibodies remain less-well characterized than their mammalian counterparts. This review aims at providing a current overview of the structure, function, development and generation of chicken antibodies. Additionally, brief but comprehensive insights into current knowledge pertaining to the immunogenetic framework and diversity-generation of the chicken immunoglobulin repertoire which have contributed to the establishment of recombinant chicken mAb-generating methods are discussed...
May 11, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28478117/development-of-an-oral-fluid-immunoassay-to-assess-past-and-recent-hepatitis-e-virus-hev-infection
#5
Nora Pisanic, Atiqur Rahman, Samir K Saha, Alain B Labrique, Kenrad E Nelson, Douglas A Granger, Steve W Granger, Barbara Detrick, Christopher D Heaney
BACKGROUND: Hepatitis E virus (HEV) infection causes significant morbidity and mortality worldwide, particularly among pregnant women. In clinical settings blood-based testing protocols are commonly used to diagnose HEV infection, but in community settings such invasive sampling can hinder study participation and limit discovery of the ecology and natural history of HEV infection. Oral fluid is a non-invasive biospecimen that can harbor pathogen-specific antibodies and has the potential to replace blood-based testing protocols...
May 3, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28472622/flow-cytometric-quantitation-of-platelet-phagocytosis-by-monocytes-using-a-ph-sensitive-dye-phrodo-se
#6
Daisuke Takahashi, Mitsuhiro Fujihara, Toru Miyazaki, Keiji Matsubayashi, Shinichiro Sato, Hiroshi Azuma, Toshiaki Kato, Shuichi Kino, Hisami Ikeda, Shigeru Takamoto, Noriyuki Sato, Toshihiko Torigoe
No abstract text is available yet for this article.
May 1, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28464989/induction-of-neonatal-tolerance-to-gfp-labeled-karyocytes-in-c57-b6-mice
#7
Roman Dovgiy, Iryna Pishel, Gennadij Butenko, Larysa Skivka
Green fluorescent protein is widely used in biological studies including parabiosis models for visualization of cellular structures and cells. However, the growing number of the data is available regarding immunogenicity of this protein, which can interfere with its use in in vivo experiments. In this study, we attempted to induce neonatal immunological tolerance to GFP-labeled karyocytes by intraperitoneal injections of B6.GFP mouse splenocytes to newborn C57/B6 mice. GFP(+) skin graft integrity was evaluated under UV light at 6weeks after skin grafting...
April 29, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28454715/a-pcr-based-method-for-quantifying-neutrophils-in-human-nasal-secretions
#8
Matthew C Morris, M Nadeem Khan, Michael E Pichichero
Neutrophil recruitment to the nasopharynx (NP) is a central event in resolution of NP-initiated microbial infections. A vigorous neutrophil response in infected tissues is also associated with the outcome of adverse tissue pathology. Therefore, differences in infection-induced tissue neutrophil numbers may correlate with pathogenesis events. Existing methods of quantifying neutrophils require evaluation of NP samples within hours of procurement as flow cytometry based cell quantification methods require live neutrophil cells...
April 25, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28434982/novel-monoclonal-antibodies-against-stx1d-and-1e-and-their-use-for-improving-immunoassays
#9
Xiaohua He, Stephanie Patfield, Reuven Rasooly, Daniela Mavrici
Shiga toxins (Stxs) are major causative agents for bloody diarrhea and hemolytic uremic syndrome, a life-threatening disease in humans. No effective treatment is available. Early detection of Stxs in clinical samples is critical for disease management. As bacteria evolve, new Stxs are produced; therefore, methods used to identify them need to be improved as well. In this study, new monoclonal antibodies (mAbs) against Stx1d and 1e were developed and used to improve a commercial Stx1 kit. Incorporation of the new mAbs into the Abraxis Stx1 kit not only increased the assay sensitivity to Stx1d, but the assay was conferred the ability to detect Stx1e, a newly identified subtype of Stx1 produced by an atypical Stx-producing bacterial strain, Enterobacter cloacae M12X01451, isolated from a clinical specimen...
April 20, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28434981/protein-transport-inhibitors-downregulate-the-expression-of-lag-3-on-regulatory-t-cells
#10
Sara Anvari, Andrea Grimbergen, Carla M Davis, George Makedonas
Modern immunologic studies demand increasing complexity because of a need to improve our understanding of the relationship between a cell's phenotype and its function. Regulatory T cells have been defined by a narrow set of phenotypic markers, however their actual functional capacity has not been determined at the single-cell level. Although the lymphocyte activation gene 3 (LAG-3; CD223) is a key marker for the identification of exhausted T cells, it may be useful also in resolving Treg subpopulations by indicating distinct functional breadths...
April 20, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28434980/thaw-and-use-target-cells-pre-labeled-with-calcein-am-for-antibody-dependent-cell-mediated-cytotoxicity-assays
#11
Shan Chung, Van Nguyen, Yuwen Linda Lin, Lynn Kamen, An Song
In vitro antibody-dependent cell-mediated cytotoxicity (ADCC) assays are routinely performed to support the research and development of therapeutic antibodies. In ADCC assays, target cells bound by the antibodies are lysed by activated effector cells following interactions between the Fc region of the bound antibody and Fcγ receptors on effector cells. Target cell lysis is typically measured by quantification of released endogenous enzymes, e.g., lactate dehydrogenase, or measurement of released exogenous labels, e...
April 20, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28433580/description-of-a-novel-multiplex-avidity-assay-for-evaluating-hpv-antibodies
#12
Allison M Brady, Elizabeth R Unger, Gitika Panicker
Limited data exists regarding antibody avidity for human papillomavirus (HPV). We describe development of a multiplex electrochemiluminescent avidity ELISA for four HPV types (HPV 6, 11, 16, 18) by adding a dissociating step to our established multiplex HPV VLP ELISA. Initial experiments exploring ammonium thiocyanate, sodium thiocyanate and guanidine hydrochloride (GuHCl) as dissociating agents identified GuHCl as most promising. Dissociation conditions with GuHCl were varied (concentration, incubation time, temperature) to select conditions with minimal impact on VLP integrity as measured with monoclonal antibodies to conformational epitopes...
April 19, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28433579/distinguishing-septic-from-normal-donors-by-detection-of-spla2-iia-from-human-plasma-using-a-microsieve-based-immunoassay
#13
Daniel R Zweitzig, Arjan G Tibbe, Mark J Kopnitsky, Kathleen Cichonski, Leon W M M Terstappen
Bloodstream infections that progress to septic shock are responsible for hundreds of thousands of deaths each year, and are associated with significant healthcare costs. Recent studies have shown that a member of the secreted phospholipase protein family, termed sPLA2-IIA, may play a role during the innate immune response to bacterial infections, and is elevated in the plasma of septic patients. In this report, the feasibility of a simple microsieve-based sPLA2-IIA detection immunoassay was explored. Microsieves containing 5μm pores were covalently coupled with a sPLA2-IIA-specific monoclonal antibody at 0...
April 19, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28414024/novel-high-throughput-cell-based-hybridoma-screening-methodology-using-the-celigo-image-cytometer
#14
Haohai Zhang, Leo Li-Ying Chan, William Rice, Nasim Kassam, Maria Serena Longhi, Haitao Zhao, Simon C Robson, Wenda Gao, Yan Wu
Hybridoma screening is a critical step for antibody discovery, which necessitates prompt identification of potential clones from hundreds to thousands of hybridoma cultures against the desired immunogen. Technical issues associated with ELISA- and flow cytometry-based screening limit accuracy and diminish high-throughput capability, increasing time and cost. Conventional ELISA screening with coated antigen is also impractical for difficult-to-express hydrophobic membrane antigens or multi-chain protein complexes...
April 14, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28392436/an-oral-administration-of-a-recombinant-anti-tnf-fusion-protein-is-biologically-active-in-the-gut-promoting-regulatory-t-cells-results-of-a-phase-i-clinical-trial-using-a-novel-oral-anti-tnf-alpha-based-therapy
#15
Einat Almon, Tawfik Khoury, Ariel Drori, Svetlana Gingis-Velitski, Sari Alon, Raul Chertkoff, Mordechai Mushkat, Yoseph Shaaltiel, Yaron Ilan
BACKGROUND: An orally administered BY-2 plant cell-expressed recombinant anti-TNF fusion protein (PRX-106) consists of the soluble form of the human TNF receptor (TNFR) fused to the Fc component of a human IgG1 domain. Aim This study aim at determining the safety and the immune modulatory effect of an oral administration of PRX-106 in humans. METHODS: Three different doses (2, 8 or 16mg/day) of PRX-106 were orally administered for five consecutive days in 14 healthy volunteered participants...
April 7, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28390927/a-novel-hemolytic-complement-sufficient-nsg-mouse-model-supports-studies-of-complement-mediated-antitumor-activity-in-vivo
#16
Mohit K Verma, Julia Clemens, Lisa Burzenski, Stephen B Sampson, Michael A Brehm, Dale L Greiner, Leonard D Shultz
Monoclonal antibodies (mAbs) have emerged as a mainstream therapeutic option against cancer. mAbs mediate tumor cell-killing through several mechanisms including complement-dependent cytotoxicity (CDC). However, studies of mAb-mediated CDC against tumor cells remain largely dependent on in vitro systems. Previously developed and widely used NOD-scid IL2rγ(null) (NSG) mice support enhanced engraftment of many primary human tumors. However, NSG mice have a 2-bp deletion in the coding region of the hemolytic complement (Hc) gene, and it is not possible to evaluate CDC activity in NSG mice...
April 6, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28390926/rhodopsin-in-plasma-from-patients-with-diabetic-retinopathy-development-and-validation-of-digital-elisa-by-single-molecule-array-simoa-technology
#17
Eva Rabing Brix Petersen, Dorte Aalund Olsen, Henry Christensen, Søren Berndt Hansen, Cramer Christensen, Ivan Brandslund
BACKGROUND: Diabetic retinopathy (DR) is the most frequent cause of blindness among younger adults in the western world. No blood biomarkers exist to detect DR. Hypothetically, Rhodopsin concentrations in blood has been suggested as an early marker for retinal damage. The aim of this study was therefore to develop and validate a Rhodopsin assay by employing digital ELISA technology, and to investigate whether Rhodopsin concentrations in diabetes patients with DR are elevated compared with diabetes patients without DR...
April 5, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28390925/proteomic-analysis-of-human-plasma-and-peripheral-blood-mononuclear-cells-in-systemic-lupus-erythematosus-patients
#18
Sara Pinna, Sara Pasella, Marta Deiana, Angela Baralla, Andrea Mannu, Antonio Giovanni Emilio Masala, Piera Veronica Pileri, Nicola Deiana, Fabrizio Scognamillo, Carlo Pala, Angelo Zinellu, Ciriaco Carru, Luca Deiana
Systemic Lupus Erythematosus (SLE) is a systemic autoimmune disease with a broad spectrum of clinical presentations and incompletely understood pathogenesis. This autoimmune disease is characterized by alterations in both the innate and adaptive immune system that lead to the loss of immunologic tolerance. In autoimmune diseases particularly in SLE, early diagnosis, flare or remission phases can be difficult to identify. Proteomics can help to find new therapeutic targets and it also could help to better understand the cellular mechanisms...
April 5, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28389175/validation-of-a-multi-analyte-panel-with-cell-bound-complement-activation-products-for-systemic-lupus-erythematosus
#19
Thierry Dervieux, John Conklin, Jo-Anne Ligayon, Leilani Wolover, Tyler O'Malley, Roberta Vezza Alexander, Arthur Weinstein, Claudia A Ibarra
BACKGROUND: We describe the analytical validation of an assay panel intended to assist clinicians with the diagnosis of systemic lupus erythematosus (SLE). The multi-analyte panel includes quantitative assessment of complement activation and measurement of autoantibodies. METHODS: The levels of the complement split product C4d bound to erythrocytes (EC4d) and B-lymphocytes (BC4d) (expressed as mean fluorescence intensity [MFI]) are measured by quantitative flow cytometry, while autoantibodies (inclusive of antinuclear and anti-double stranded DNA antibodies) are determined by immunoassays...
April 5, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28389174/a-biotin-drug-extraction-and-acid-dissociation-bead-procedure-to-eliminate-matrix-and-drug-interference-in-a-protein-complex-anti-drug-antibody-ada-isotype-specific-assay
#20
Hongmei Niu, Thomas Klem, Jinsong Yang, Yongchang Qiu, Luying Pan
Monitoring anti-drug antibody (ADA) responses in patients receiving protein therapeutics treatment is an important safety assessment for regulatory agencies, drug manufacturers, clinicians and patients. Recombinant human IGF-1/IGFBP-3 (rhIGF-1/rhIGFBP-3) is a 1:1 formulation of naturally occurring protein complex. The individual IGF-1 and IGFBP-3 proteins have multiple binding partners in serum matrix with high binding affinity to each other, which presents challenges in ADA assay development. We have developed a biotin-drug extraction with acid dissociation (BEAD) procedure followed by an electrochemiluminescence (ECL) direct assay to overcome matrix and drug interference...
April 4, 2017: Journal of Immunological Methods
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