Dataset Information

Immune repertoire after immunization as seen by next generation sequencing and proteomics

ABSTRACT: We study the repertoire of immunoglobulin sequences after immunization in a cohort of rats. Animals were immunized with dinitrophenol modified KLH or with recombinant human HuD. In the immune repertoire, and also in the matching proteomics data, we observe that the animals produce a repertoire that has many shared motifs for those immunized with the same antigen. Cluster analysis allows the samples to be segregated according to the immunogen used.

OTHER RELATED OMICS DATASETS IN: PRJNA386462

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Rattus Norvegicus (rat)

TISSUE(S): Blood Serum

SUBMITTER: Martijn van Duijn

LAB HEAD: Theo Luider

PROVIDER: PXD006484 | Pride | 2017-10-04

REPOSITORIES: Pride

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Publications

Immune responses are characterized by specific shared immunoglobulin peptides that can be detected by proteomic techniques.

VanDuijn Martijn M MM Dekker Lennard J M LJ Zeneyedpour L L Smitt Peter A E Sillevis PA Luider Theo M TM

The Journal of biological chemistry 20100708 38

In the adaptive immune response, immunoglobulins develop that bind specifically to the antigens to which the organism was exposed. Immunoglobulins may bind to known or unknown antigens in a variety of diseases and have been used in the past to identify novel antigens for use as a biomarker. We propose that the immunoglobulins themselves could also be used as biomarkers in antibody-mediated disease. In this proteomic study, rats were immunized with one of two purified antigens, and immunoglobulin ...[more]

PMID: 20615873

Publication: 1/2

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Project description:The discovery of hepatitis C virus (HCV) in 1989 revealed the virus as the etiology of 40%-90% of the “essential” mixed cryoglobulinemia, where immune complex forms deposit called cryoprecipitate at temperatures below 37 °C. Cryoprecipitate constitutes monoclonal IgM and polyclonal IgG, some of which has reactivity against HCV core and NS3 epitopes. Resultant immune complex is considered entrapped on microvascular endothelium via C1q receptor, leading to complement activation and organ injury presenting predominantly as dermopathy, peripheral neuropathy and nephropathy. However, currently little is known on whether auto-reactive, cold-precipitating IgG components enriched in cryoprecipitate may play some role on the deposition of immune complex and subsequent complement-mediated injury of specific organs. Recently, with the advent of high-throughput immune repertoire sequencing and mass spectrometry, technical feasibility is growing to delineate antibodies of interest and their sequences directly from serum. To date, vast majority of studies actually utilized the antigen column for affinity purification of antibodies of interest, although this strategy is not applicable to disease entity with unknown antigen involvement. In such cases, disease-specific and organ-specific immune deposits may be a good alternative source of etiological antibodies. Herein, targeting HCV cryoglobulinemic vasculitis as a model, we conducted a proof-of-concept study aiming at characterizing the IgG components most prone to cryoprecipitation. To this end, we longitudinally studied one patient with cryoglobulinemic vasculitis with chronic HCV infection. After obtaining informed consent, cryoprecipitate and supernatant were separated from peripheral blood sample. Fab fragments from Protein G-purified IgG were recovered after papain digestion for isobaric tags for relative and absolute quantification (iTRAQ)-based quantitative proteomics. Simultaneously, total RNA was isolated from peripheral blood, and immunoglobulin heavy chain variable region (IGHV) was PCR-amplified with unique molecular identifier (UMI) strategy to construct a personal IGHV sequence library of immunoglobulin variable region. Sequencing output from MiSeq was bioinformatically converted into mass spectrometry database. Search was performed using MaxQuant software.

Dataset Information

Immune repertoire after immunization as seen by next generation sequencing and proteomics

Publications

Immune responses are characterized by specific shared immunoglobulin peptides that can be detected by proteomic techniques.

Similar Datasets

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