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Frontiers in Immunology 2021Most vaccines require multiple doses to induce long-lasting protective immunity in a high frequency of vaccines, and to ensure strong both individual and herd immunity.... (Review)
Review
Most vaccines require multiple doses to induce long-lasting protective immunity in a high frequency of vaccines, and to ensure strong both individual and herd immunity. Repetitive immunogenic stimulations not only increase the intensity and durability of adaptive immunity, but also influence its quality. Several vaccine parameters are known to influence adaptive immune responses, including notably the number of immunizations, the delay between them, and the delivery sequence of different recombinant vaccine vectors. Furthermore, the initial effector innate immune response is key to activate and modulate B and T cell responses. Optimization of homologous and heterologous prime/boost vaccination strategies requires a thorough understanding of how vaccination history affects memory B and T cell characteristics. This requires deeper knowledge of how innate cells respond to multiple vaccine encounters. Here, we review how innate cells, more particularly those of the myeloid lineage, sense and respond differently to a 1st and a 2nd vaccine dose, both in an extrinsic and intrinsic manner. On one hand, the presence of primary specific antibodies and memory T cells, whose critical properties change with time after priming, provides a distinct environment for innate cells at the time of re-vaccination. On the other hand, innate cells themselves can exert enhanced intrinsic antimicrobial functions, long after initial stimulation, which is referred to as trained immunity. We discuss the potential of trained innate cells to be game-changers in prime/boost vaccine strategies. Their increased functionality in antigen uptake, antigen presentation, migration, and as cytokine producers, could indeed improve the restimulation of primary memory B and T cells and their differentiation into potent secondary memory cells in response to the boost. A better understanding of trained immunity mechanisms will be highly valuable for harnessing the full potential of trained innate cells, to optimize immunization strategies.
Topics: Adaptive Immunity; Animals; Antibodies; Antibody Specificity; Humans; Immunity, Humoral; Immunity, Innate; Immunization Schedule; Immunization, Secondary; Immunologic Memory; T-Lymphocytes; Vaccination; Vaccines
PubMed: 33763063
DOI: 10.3389/fimmu.2021.612747 -
Pharmacology 2022
Topics: Antibody Specificity; Immunoglobulin A
PubMed: 35358972
DOI: 10.1159/000524041 -
International Journal of Molecular... Apr 2023Myoglobin (Mb) is the main constituent of vertebrate skeletal muscle and myocardium and plays an essential role in oxygen binding, storage, transport, and earliest...
Myoglobin (Mb) is the main constituent of vertebrate skeletal muscle and myocardium and plays an essential role in oxygen binding, storage, transport, and earliest disease diagnosis. This study focuses on preparing the novel recombinant rabbit anti-Mb monoclonal antibody and applying it to a diagnosis of Mb deposition in rhabdomyolysis-associated acute kidney injury (RM-AKI). The full-length coding sequence of rat Mb was cloned and expressed, and the high-quality and titer rabbit anti-Mb polyclonal antibodies were produced by the immunogen His-Mb fusion protein. A new hybridoma cell was obtained by hybridoma screening technology. With the help of DNA sequencing and a molecular clonal, anti-Mb monoclonal antibody heavy and light chains expression plasmid was constructed. Finally, the recombinant rabbit anti-Mb monoclonal antibody with extraordinarily high affinity (K = 1.21 pM) was obtained. Meanwhile, it had broad species reactivity (mouse, rat, human, and horse) and good tissue specificity (skeletal muscle and myocardium). It also had a very good performance in western blotting, immunohistochemistry, and immunofluorescence assay to detect the Mb level in the kidney, myocardium, and skeletal muscle of RM-AKI. This study will be significantly helpful for Mb-associated disease diagnosis, and pathogenesis exploration, and further may act as a neutralizing antibody for disease treatment.
Topics: Rabbits; Humans; Mice; Rats; Animals; Horses; Myoglobin; Rhabdomyolysis; Acute Kidney Injury; Kidney; Antibodies, Monoclonal; Antibody Specificity
PubMed: 37175528
DOI: 10.3390/ijms24097822 -
Journal of Visualized Experiments : JoVE Aug 2017Post-translational modifications (PTMs) on histone proteins are widely studied for their roles in regulating chromatin structure and gene expression. The mass production...
Post-translational modifications (PTMs) on histone proteins are widely studied for their roles in regulating chromatin structure and gene expression. The mass production and distribution of antibodies specific to histone PTMs has greatly facilitated research on these marks. As histone PTM antibodies are key reagents for many chromatin biochemistry applications, rigorous analysis of antibody specificity is necessary for accurate data interpretation and continued progress in the field. This protocol describes an integrated pipeline for the design, fabrication and use of peptide microarrays for profiling the specificity of histone antibodies. The design and analysis aspects of this procedure are facilitated by ArrayNinja, an open-source and interactive software package we recently developed to streamline the customization of microarray print formats. This pipeline has been used to screen a large number of commercially available and widely used histone PTM antibodies, and data generated from these experiments are freely available through an online and expanding Histone Antibody Specificity Database. Beyond histones, the general methodology described herein can be applied broadly to the analysis of PTM-specific antibodies.
Topics: Antibodies; Antibody Specificity; Histones; Lysine; Peptides; Protein Array Analysis; Protein Processing, Post-Translational; Robotics; Software; Threonine
PubMed: 28809825
DOI: 10.3791/55912 -
Journal of Molecular Biology May 2023Monoclonal antibody (mAb)-based biologics are well established treatments of cancer. Antibody discovery campaigns are typically directed at a single target of interest,...
Monoclonal antibody (mAb)-based biologics are well established treatments of cancer. Antibody discovery campaigns are typically directed at a single target of interest, which inherently limits the possibility of uncovering novel antibody specificities or functionalities. Here, we present a target-unbiased approach for antibody discovery that relies on generating mAbs against native target cell surfaces via phage display. This method combines a previously reported method for improved whole-cell phage display selections with next-generation sequencing analysis to efficiently identify mAbs with the desired target cell reactivity. Applying this method to multiple myeloma cells yielded a panel of >50 mAbs with unique sequences and diverse reactivities. To uncover the identities of the cognate antigens recognized by this panel, representative mAbs from each unique reactivity cluster were used in a multi-omic target deconvolution approach. From this, we identified and validated three cell surface antigens: PTPRG, ICAM1, and CADM1. PTPRG and CADM1 remain largely unstudied in the context of multiple myeloma, which could warrant further investigation into their potential as therapeutic targets. These results highlight the utility of optimized whole-cell phage display selection methods and could motivate further interest in target-unbiased antibody discovery workflows.
Topics: Humans; Antibodies, Monoclonal; Antibody Specificity; Antigens; Cell Adhesion Molecule-1; Cell Surface Display Techniques; Multiomics; Multiple Myeloma; Peptide Library
PubMed: 37019174
DOI: 10.1016/j.jmb.2023.168085 -
Immunity, Inflammation and Disease Dec 2021The efficacy assessment of human anti-IgE monoclonal antibodies (mAbs) in animal models before clinical trials is hampered due to the lack of cross-reactivity of...
BACKGROUND
The efficacy assessment of human anti-IgE monoclonal antibodies (mAbs) in animal models before clinical trials is hampered due to the lack of cross-reactivity of anti-IgE mAbs between species.
OBJECTIVE
We developed CRE-DR (an anti-dog IgE monoclonal antibody), an anti-IgE mouse mAb that recognizes canine and human IgE, and then examined its IgE specificity and cross-reactivity between three animal and human species.
METHODS
After mouse immunization with a synthetic peptide derived from canine IgE ( NTNDWIEGETYYC ), we generated a hybridoma producing CRE-DR. The CRE-DR purified from the ascites of hybridoma-inoculated mice was used for ELISA and Western blot analysis to examine reactivity to dog, human, and rodent IgEs as well as recombinant bovine serum albumin (BSA)-conjugated to canine, human, and rodent IgE amino acid peptides corresponding to the immunizing sequence. We then performed enzyme-linked immunosorbent assays (ELISAs) for dog IgE using sera from dogs with atopic dermatitis (AD) after inhibition with canine IgE and IgG. The amino acid sequence recognized by CRE-DR was identified by ELISA using synthetic peptides.
RESULTS
CRE-DR is a monoclonal mouse IgG1κ specific for dog IgE, and the ELISA values in atopic dog sera were inhibited by dog IgE, but not dog IgG. The binding of CRE-DR to human IgE was relatively maintained, but not to rodent IgEs, which results were confirmed with the BSA-conjugated IgE peptides of the various species. The CRE-DR reactivity was supported by the comparison of amino acid sequence of CRE-DR epitope, DWIEGETYYC, in dog IgE; one, two, and three amino acids were substituted in the human, rat, and mouse IgE epitopes, respectively.
CONCLUSIONS AND CLINICAL RELEVANCE
CRE-DR is a mAb cross-reactive to dog and human IgEs, which can allow the use of a dog model of allergy to test the efficacy of a CRE-DR-derived anti-IgE therapeutic mAb before human clinical trials.
Topics: Animals; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Antibody Specificity; Cross Reactions; Dogs; Humans; Immunoglobulin E; Mice; Rats
PubMed: 34533288
DOI: 10.1002/iid3.531 -
IgG Donor-Specific Anti-Human HLA Antibody Subclasses and Kidney Allograft Antibody-Mediated Injury.Journal of the American Society of... Jan 2016Antibodies may have different pathogenicities according to IgG subclass. We investigated the association between IgG subclasses of circulating anti-human HLA antibodies...
Antibodies may have different pathogenicities according to IgG subclass. We investigated the association between IgG subclasses of circulating anti-human HLA antibodies and antibody-mediated kidney allograft injury. Among 635 consecutive kidney transplantations performed between 2008 and 2010, we enrolled 125 patients with donor-specific anti-human HLA antibodies (DSA) detected in the first year post-transplant. We assessed DSA characteristics, including specificity, HLA class specificity, mean fluorescence intensity (MFI), C1q-binding, and IgG subclass, and graft injury phenotype at the time of sera evaluation. Overall, 51 (40.8%) patients had acute antibody-mediated rejection (aABMR), 36 (28.8%) patients had subclinical ABMR (sABMR), and 38 (30.4%) patients were ABMR-free. The MFI of the immunodominant DSA (iDSA, the DSA with the highest MFI level) was 6724±464, and 41.6% of patients had iDSA showing C1q positivity. The distribution of iDSA IgG1-4 subclasses among the population was 75.2%, 44.0%, 28.0%, and 26.4%, respectively. An unsupervised principal component analysis integrating iDSA IgG subclasses revealed aABMR was mainly driven by IgG3 iDSA, whereas sABMR was driven by IgG4 iDSA. IgG3 iDSA was associated with a shorter time to rejection (P<0.001), increased microcirculation injury (P=0.002), and C4d capillary deposition (P<0.001). IgG4 iDSA was associated with later allograft injury with increased allograft glomerulopathy and interstitial fibrosis/tubular atrophy lesions (P<0.001 for all comparisons). Integrating iDSA HLA class specificity, MFI level, C1q-binding status, and IgG subclasses in a Cox survival model revealed IgG3 iDSA and C1q-binding iDSA were strongly and independently associated with allograft failure. These results suggest IgG iDSA subclasses identify distinct phenotypes of kidney allograft antibody-mediated injury.
Topics: Allografts; Antibody Specificity; Female; HLA Antigens; Humans; Immunoglobulin G; Kidney Diseases; Kidney Transplantation; Male; Middle Aged; Postoperative Complications; Tissue Donors; Transplantation Immunology
PubMed: 26293822
DOI: 10.1681/ASN.2014111120 -
Medecine Sciences : M/S Dec 2019In 2019, monoclonal antibodies are a worldwide annual business worth of more than 100 billions USD (i.e., about 90 billions €). In addition to their use in the... (Review)
Review
In 2019, monoclonal antibodies are a worldwide annual business worth of more than 100 billions USD (i.e., about 90 billions €). In addition to their use in the clinics, monoclonal antibodies are also used for diagnosis and remain highly valuable tools for academic basic and translational research. Forty-four years after the seminal publication of Georges Köhler and César Milstein, dozens of meetings and seminars focusing on various aspects of mAbs are held annually all around the world. But forty-four years later, the scientific works and efforts that have made possible this scientific breakthrough are gradually forgotten and, for many, monoclonal antibodies are no more than a multi-million USD business alike any other big business, guided by financial markets and the results of on-going clinical trials… Time has now come for acknowledging and paying tribute to all these scientists involved in basic research, to these researchers passionate about science, some famous, some forgotten, scattered all over the world. They explored during the 20 century the frontiers of unknown and generated a knowledge that allowed the emergence of a technique that translated finally into what is one of the greatest therapeutic revolution of the modern era.
Topics: Animals; Antibodies, Monoclonal; Antibody Diversity; Antibody Specificity; Biomedical Research; Drug Industry; Exploratory Behavior; History, 20th Century; History, 21st Century; Humans
PubMed: 31903896
DOI: 10.1051/medsci/2019222 -
Polski Przeglad Chirurgiczny Feb 2018The human leukocyte antigen (HLA) system plays an important role in the acceptance of renal graft. Long and better graft survival has been reported in patients with... (Review)
Review
The human leukocyte antigen (HLA) system plays an important role in the acceptance of renal graft. Long and better graft survival has been reported in patients with HLA-identical siblings and a nonreactive cytotoxicity assay (CDC). New methods of HLA-typing and anti-HLA antibody detection techniques such as flow cytometry, solid-phase immunoassays, or antigen bead assays have further improved the outcomes of renal transplant recipients. In the present review, the explicit details of these methodologies are discussed in detail.
Topics: Antibody Specificity; Flow Cytometry; Graft Rejection; Graft Survival; HLA Antigens; Histocompatibility; Histocompatibility Testing; Humans; Isoantibodies; Kidney Transplantation
PubMed: 29513250
DOI: 10.5604/01.3001.0011.5959 -
Frontiers in Immunology 2018Antigens in particulate form have distinct immunologic properties relative to soluble antigens. An understanding of the mechanisms and functional consequences of the... (Review)
Review
Antigens in particulate form have distinct immunologic properties relative to soluble antigens. An understanding of the mechanisms and functional consequences of the distinct immunologic pathways engaged by these different forms of antigen is particularly relevant to the design of vaccines. It is also relevant regarding the use of therapeutic human proteins in clinical medicine that have been shown to aggregate, and perhaps as a result, elicit autoantibodies.
Topics: Animals; Antibody Formation; Antibody Specificity; Antigen-Presenting Cells; Antigens; B-Lymphocytes; Epitopes; Humans; Immune Tolerance; Immunity; Immunogenicity, Vaccine; Solubility; Vaccines
PubMed: 29619034
DOI: 10.3389/fimmu.2018.00598