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Journal of Immunological Methods

V V Krishnan, Senthamil R Selvan, Nishanth Parameswaran, Neeraja Venkateswaran, Paul A Luciw, Kodumudi S Venkateswaran
Advances in high-throughput proteomic approaches have provided substantial momentum to novel disease-biomarker discovery research and have augmented the quality of clinical studies. Applications based on multiplexed microsphere suspension array technology are making strong in-roads into the clinical diagnostic/prognostic practice. Conventional proteomic approaches are designed to discover a broad set of proteins that are associated with a specific medical condition. In comparison, multiplex microsphere immunoassays use quantitative measurements of selected set(s) of specific/particular molecular markers such as cytokines, chemokines, pathway signaling or disease-specific markers for detection, metabolic disorders, cancer, and infectious agents causing human, plant and animal diseases...
July 9, 2018: Journal of Immunological Methods
Ryu Nakano, Kazuhide Nakagaki, Yuko Itoh, Utako Seino, Takahiro Ueda, Ryushi Tazawa, Nobutaka Kitamura, Takahiro Tanaka, Koh Nakata
During a clinical trial of a Saccharomyces cerviciae-derived recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF), sargramostim, in patients with autoimmune pulmonary alveolar proteinosis (aPAP), we conducted a pharmacokinetic study of single-dose sargramostim inhalation. Several problems were encountered whereby sargramostim formed an immune-complex with GM-CSF autoantibodies (GMAbs) immediately after entering the body; thus, we could not measure the concentration of sargramostim using a commercial high sensitivity enzyme-linked immunosorbent assay (ELISA)...
July 9, 2018: Journal of Immunological Methods
Meseret Habtamu, Markos Abebe, Abraham Aseffa, Anne Margarita Dyrhol Riise, Anne Spurkland, Greger Abrahamsen
There is a lack of suitable correlates of immune protection against Mycobacterium tuberculosis (Mtb) infection. T cells and monocytes play key roles in host immunity against Mtb. Thus, a method that allows assessing their interaction would contribute to the understanding of immune regulation in tuberculosis (TB). We have established imaging flow cytometer (IFC) based in vitro assay for the analysis of early events in T cell-monocyte interaction, upstream of cytokine production and T cell proliferation. This was achieved through short term stimulation of peripheral blood mononuclear cells (PBMC) from healthy Norwegian blood donors with Mycobacterium bovis Bacille Calmette-Guérin (BCG)...
July 4, 2018: Journal of Immunological Methods
Francesca Necchi, Allan Saul, Simona Rondini
Increasing awareness of Salmonella Paratyphi A's contribution to enteric fever episodes throughout Asia has led to the development of new S. Paratyphi A vaccines. Assays are needed to measure functional antibodies elicited by the new vaccine candidates to assess their immunogenicity and potential protective capacities. Serum bactericidal assay (SBA) is the method of choice to measure functional antibody titers against various bacterial pathogens, but it is rarely been used for large dataset and clinical samples because it is time consuming and labor-intensive...
June 30, 2018: Journal of Immunological Methods
Ahmed M Alshehri, Thilini Mendis, Michael W Jackson
Recent studies have postulated the presence of functional autoantibodies (Abs) against L-type voltage gated calcium channels (VGCCs) in the serum of patients with type 1 diabetes, with various proposed physiological consequences, both islet cell associated and extra-glandular. Arguably, the most potentially damaging effect reported for these Abs is induction of apoptosis in pancreatic beta (β) cells, yet a convincing pathogenic mechanism remains to be demonstrated. In the current study, we report an assay of reactive oxygen species (ROS) stress induction in the rat insulinoma cell line Rin A12, as determined by 2', 7'-Dichlorofluorescein diacetate (DCF-DA) fluorescence detection by flow cytometry...
June 22, 2018: Journal of Immunological Methods
Igor C Borges, Dafne C Andrade, Nina Ekström, Camilla Virta, Merit Melin, Annika Saukkoriipi, Maija Leinonen, Olli Ruuskanen, Helena Käyhty, Cristiana M Nascimento-Carvalho
The aim of this study was to compare the results of serological assays using pneumococcal proteins or polysaccharides for the detection of pneumococcal infection in childhood pneumonia. Serological assays measured IgG against eight pneumococcal proteins (Ply,CbpA,PspA1,PspA2,PcpA,PhtD,StkP-C,PcsB-N), C-polysaccharide [in the whole study population, n = 183], or 19 pneumococcal capsular polysaccharides (1,2,4,5,6B,7F,8,9 V,10A,11A,12F,14,15B,17F,18C,19F,20,23F,33F) [only in a subgroup of patients, n = 53] in paired serum samples of children aged <5 years-old hospitalized with clinical and radiological diagnosis of community-acquired pneumonia...
June 20, 2018: Journal of Immunological Methods
Maria A V Willrich, Bruna D Andreguetto, Meera Sridharan, Fernando C Fervenza, Linda J Tostrud, Paula M Ladwig, Ann M Rivard, MeLea D Hetrick, Ryan N Olson, Sandra C Bryant, Melissa R Snyder, David L Murray
BACKGROUND: Eculizumab (ECU) blocks complement C5 cleavage, preventing the formation of C5a and the cytolytic effects of the membrane attack complex. The presence of ECU in blood impacts routine complement tests used to monitor treatment. METHODS: Residual serum samples with normal total complement (CH50) and residual citrate plasma with normal PT/APTT were spiked with ECU at varied concentrations ranging from 25 to 600 μg/mL. In addition, seventy-one samples from patients on ECU were obtained...
June 20, 2018: Journal of Immunological Methods
Karnail Singh, Bishal Marasini, Xuemin Chen, Paul Spearman
Ebolaviruses are highly virulent pathogens that cause Ebola viral disease (EVD). Data from non-human primate (NHP) models and from human survivors of EVD suggest that anti-Ebola antibodies play an integral role in protection. Antibody-dependent cell-mediated cytotoxicity (ADCC) is a potential mechanism through which anti-Ebola antibodies may mediate protection. We developed a robust Ebola-specific ADCC assay for use in ongoing trials of Ebola vaccines. Stable cell lines for inducible Zaire ebolavirus glycoprotein (EBOV GP) expression were developed to provide a uniform source of target cells in the assay, and were combined with an existing human natural killer (NK) cell line as the effector cell...
June 15, 2018: Journal of Immunological Methods
Ida Marie Rundgren, Øystein Bruserud, Anita Ryningen, Elisabeth Ersvær
INTRODUCTION: Monocytes are important for innate immunity and include the classical (CD14bright CD16negative ), intermediate (CD14bright CD16dim ) and non-classical (CD14dim CD16bright ) monocyte subsets. The quantification of these functionally different subsets in peripheral blood may become useful for diagnosis and follow-up in human diseases. The aim of the present study was to investigate how different pre-analytical parameters influence analysis of monocyte subsets in peripheral blood samples...
June 12, 2018: Journal of Immunological Methods
Girja S Shukla, Yu-Jing Sun, Stephanie C Pero, Giselle S Sholler, David N Krag
The aim of this preclinical study was to evaluate T7 bacteriophage as a nanoparticle platform for expression of neoantigens that could allow rapid generation of vaccines for potential studies in human cancer patients. We have generated recombinant T7 phage vaccines carrying neoepitopes derived from mutated proteins of B16-F10 melanoma tumor cells. With the single mutated amino acid (AA) centered, peptides were expressed on the outer coat of T7 phage. All peptides with 11 and 34 AAs were successfully expressed...
June 12, 2018: Journal of Immunological Methods
Jon Cantero, Meritxell Genescà
In the field of sexually transmitted infections (STI), the cervicovaginal explant (CVEx) model, not only provides the opportunity to study the different immunological arms present in these tissues under steady state conditions, but also their response against ex vivo infection with relevant pathogens. The methodology associated to the establishment of the HIV infection model in the cervicovaginal tissue was described in detail by Grivel et al. earlier (Grivel and Margolis, 2009). With this model as a foundation, we illustrate different approaches to obtain a large number of immunological readouts from a single piece of tissue, thus maximizing the immunological output obtained...
June 9, 2018: Journal of Immunological Methods
Claudia Mroz, Kristina M Schmidt, Sven Reiche, Martin H Groschup, Martin Eiden
Rift Valley fever virus is a mosquito-borne virus which is associated with acute hemorrhagic fever leading to large outbreaks among ruminants and humans in Africa and the Arabian Peninsula. RVFV circulates between mosquitoes, ruminants, camels and humans, which requires divergent amplification and maintenance strategies that have not been fully explored on the cellular and molecular level. We therefore assessed monoclonal antibodies for their applicability to monitor the expression pattern and kinetics of viral proteins in different RVFV infected cell species...
June 9, 2018: Journal of Immunological Methods
Lan Zhao, Wei Wei, Yongdong Huang, Rongyue Zhang, Kai Zhu, Dongxia Hao, Zhiguo Su, Guanghui Ma
Protein A chromatography has been a popular method for purification of therapeutic monoclonal antibodies (mAb). Protein A chromatographic media using alkali-resistant rProtein A ligands from site-directed coupling method have been pursued for both high dynamic capacities and excellent stabilities. However, the mechanism of rProtein A leaking under cleaning-in-place (CIP) conditions is not very clear and difficulties have been commonly encountered when improving the media's chromatographic performance. We investigated the chromatographic performance of site-directed coupled rProtein A chromatographic media during CIP procedure...
June 9, 2018: Journal of Immunological Methods
Premrutai Thitilertdecha, Pornpichaya Suwannachod, Poonsin Poungpairoj, Varangkana Tantithavorn, Ladawan Khowawisetsut, Palanee Ammaranond, Nattawat Onlamoon
CD4 immunotherapy is potentially useful in immune reconstitution of CD4+ T cells for HIV-infected patients. Transfusion of anti-CD3/28 expanded CD4+ T cells is also proved to be safe and effective in both SIV-infected macaques and HIV-infected patients. However, there is no such standardized and practical protocol available for cell production in order to use in clinics. This study thus aimed to develop a closed-culture system for in vitro CD4+ T lymphocyte expansion by using a commercially available GMP-grade culture bag and anti-CD3/28 activation...
June 9, 2018: Journal of Immunological Methods
Ming-Shou Hsieh, Jie-Long He, Tzong-Yuan Wu, Rong-Huay Juang
A bi-cistronic baculovirus expression vector was constructed to facilitate the expression, detection, and isolation of the hemagglutinin (HA) fragment HA1 of H6N1 avian influenza virus (AIV) in an insect and a culture of its cells. In this construct, the GP67sp signal peptide promoted the secretion of the recombinant protein into the culture medium, and improved protein expression and purification. Enhanced green fluorescent protein, co-expressed through an internal ribosome entry site, served as a visible reporter for protein expression detection...
June 9, 2018: Journal of Immunological Methods
Phyllis A Rees, R Joel Lowy
Cell lines stably transfected with genes responding to Type I interferons (IFN) are potentially a useful alternative to enzyme linked immuo-assays (ELISAs) or assays based on resistance of a test cell line to virus infection using cell death or infection endpoints. Increasingly available are a variety of commercial cell lines developed for reporter gene assays (RGAs) which are responsive to IFN exposure. These cells produce a soluble gene product which can be readily quantified using multiwell plate spectrophotometers or luminometers...
June 9, 2018: Journal of Immunological Methods
Jar-How Lee
No abstract text is available yet for this article.
May 16, 2018: Journal of Immunological Methods
John Lowe, Eric Wakshull, Theresa Shek, Anan Chuntharapai, Rebecca Elliott, Jeriza Rusit, Mauricio Maia
Ranibizumab (Lucentis®), a humanized antigen-binding fragment (Fab) monoclonal antibody that blocks VEGF-A activity is currently approved for the treatment of several retinal degenerative diseases. The assessment of drug pharmacokinetics (PK) is essential for evaluating exposure as it relates to drug safety and efficacy. For drugs administered intravitreally, systemic drug levels during the course of clinical studies are typically 100 to 1000-fold lower than those of similar therapeutics dosed intravenously, posing a significant bioanalytical challenge for PK measurements...
May 14, 2018: Journal of Immunological Methods
Toshihiro Watanabe, Seiichi Hashida
Immune complex transfer enzyme immunoassay (ICT-EIA) is one of the technologies which enables ultrasensitive measurements of protein biomarkers. The ICT-EIA uses two types of beads and sandwich-shaped immune complexes are transferred from the 1st bead to the 2nd bead in the assay. The purpose of the study is to reveal the reason why the ICT-EIA achieves ultrasensitive measurements by making a detailed comparison between conventional sandwich enzyme immunoassay (Sand-EIA) and ICT-EIA. ICT-EIAs for cytokines were developed and the sensitivities were compared with the sandwich EIAs...
August 2018: Journal of Immunological Methods
Adam M Swartz, Elizabeth Reap, Pamela Norberg, Robert Schmittling, Sylvia Janetzki, Luis Sanchez-Perez, John H Sampson
The ELISpot assay prevails as one of the most sensitive and meaningful assays for the detection of antigen-specific, effector immune responses. Acquisition of cellular analyte for ELISpot analysis is typically not problematic when derived from tissues enriched in lymphocytes (e.g., lymphoid organs and blood); however, cell processing becomes more difficult when lymphocytes represent only a very minor population relative to the source tissue, especially when the source tissue is in limited supply (e.g., small mouse tumors)...
August 2018: Journal of Immunological Methods
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