Autoimmune peptide antigen reactivity in normal B6 mice Pepperchip 2.0
ABSTRACT: The aim of the array was to determine the B6 mouse sera IgG and IgM reactivity profile to linear peptide epitopes. Pooled sera from three 6-9 month old B6 mice was diluted at 1:200 for IgG specific analysis or 1:1000 for IgM specific analysis and incubated on a PEPperPRINT peptide microarray platform printed with peptide autoantigens Pooled sera from three 6-9 month old B6 mice was profiled for IgG or IgM-specific analysis of autoantibody reactivity to peptide auto-epitopes printed in duplicate on a PepperPrint Chip.
Project description:The aim of the study was to determine B6 and BXD2 mouse sera IgG and IgM reactivity to linear peptide epitopes at different ages. Serum from B6 or BXD2 mice was diluted at 1:200 for IgG-specific analysis or 1:1000 for IgM specific analysis and incubated on a PEPperPRINT peptide microarray platform printed with peptide autoantigens. 80 pre-selected peptides based on a previous screen of pooled mouse serum BXD2 against the PEPperCHIP® Autoimmunity Microarray with 2,733 linear B-cell epitopes were printed in duplicate in 16 copies on a custom PEPperCHIP® Peptide Microarray. Flag (DYKDDDDKGG) and HA (YPYDVPDYAG) control peptides (10 spots each control) were randomly distributed in each array copy as controls. Sera from 2, 5, or 9 month old B6 or BXD2 mice was profiled for anti-mouse IgG or IgM-specific analysis of autoantibody reactivity to the peptide auto-epitopes.
Project description:The aim of the array was to determine the BXD2 mouse sera IgG and IgM reactivity profile to linear peptide epitopes. Pooled sera from three 6-9 month old BXD2 mice was diluted at 1:200 for IgG specific analysis or 1:1000 for IgM specific analysis and incubated on a PEPperPRINT peptide microarray platform printed with peptide autoantigens In this study, pooled sera from three 6-9 month old BXD2 mice was profiled for IgG and IgM specific autoantibody reactivity to peptide auto-epitopes printed in duplicate on a PepperPrint Chip.
Project description:The aim of the study was to determine autoimmune BXD2 mouse sera reactivity to linear peptide epitopes from the Immune Epitope Database. Pooled sera from six 8-10 month old BXD2 mice was diluted at 1:1000 and incubated on a PEPperPRINT peptide microarray platform printed with peptide autoantigens. In this study, pooled sera from six 8-10 month old BXD2 mice was profiled for anti-mouse IgG (H+L) analysis of autoantibody reactivity to peptide auto-epitopes printed in duplicate on a PepperPrint Chip.
Project description:Citrullinated and unmodified peptides (>95% purity, ProImmune AB) were immobilized onto a chemically modified glass slide, sera from RA patients and healthy controls were applied into the reactions sites and fluorescence intensity after incubation with anti-human IgG antibody was acquired in a laser scanner. Final results for each citrullinated peptide were calculated by subtracting the intensity values of corresponding arginine containing control peptide from citrullinated peptide for all RA patients and controls.
Project description:The aim of the study was to test the hypothesis that peptide array reactivity will reveal patterns that can be associated with poor or effective neutralization response to seasonal influenza vaccine We diluted patient sera 1:80 and incubated dilutions on a streptavidin-coated glass surface that was printed with biotinylated peptides derived from seasonal strains of influenza hemagglutinin included in the 2008/2009 trivalent influenza vaccine In this study, serum from 76 study participants, subsetted into young (≤30 years old) and elderly (≥60 years old) categories were profiled for IgG/IgM antibody reactivity to influenza hemagglutinin peptudes. Using multivariate linear regression and correcting for multiple hypothesis testing by cross-validation, ee found that patterns of peptide reactivity associated with a poor or effective neutralization response to H1N1 and B strains of influenza.
Project description:Leptospirosis is the most widespread zoonotic disease in the world. The lack of an adequate laboratory test is a major barrier for the diagnosis, especially during the early stages of illness, when antibiotic therapy is most effective. Therefore, there is a critical need for an efficient diagnostic test for this life threatening disease. In order to identify new targets that could be used as diagnostic makers for leptopirosis, we constructed a protein microarray chip comprising 61% of Leptospira interrogans proteome and investigated the IgG response against leptospiral antigens from 274 individuals, including 80 acute-, 80 convalescent-phase patients and 114 healthy control subjects from regions with endemic, high endemic and no endemic transmission of leptospirosis. A nitrocellulose line blot assay was performed to validate the accuracy of the protein microarray results. We found 16 antigens that can discriminate between acute cases and healthy individuals from a region with high endemic transmission of leptospirosis, and 18 antigens that distinguish convalescent cases. Some of the antigens identified in this study, such as LipL32, the non-identical domains of the Lig proteins, GroEL and Loa22 are already known to be recognized by sera from human patients, thus serving as a proof-of-concept for the serodiagnostic antigen discovery approach. Several novel antigens were identified, including the hypothetical protein LIC10215 which showed good sensitivity and specificity rates for both acute- and convalescent-phase patients. Our study is the first large-scale evaluation of immunodominant antigens associated with naturally acquired leptospiral infection and novel as well as known serodiagnostic leptospiral antigens that are recognized by antibodies in the sera of leptospirosis cases were identified. The novel antigens identified here may have potential use in both the development of new tests and the improvement of currently available assays for diagnosing this neglected tropical disease. Further research is needed to assess the accuracy of these antigens in more appropriate diagnostic platforms. Antibody profiling was peformed on sera from infected and non-infected subjects to investigate the IgG response against leptospiral antigens. These samples comprised 80 acute-, 80 convalescent-phase patients and 114 healthy control subjects, including 29 subjects from United States (non-endemic area), 35 blood donors from Salvador/Brazil (endemic area) and 50 healthy individuals from region with high endemicity of leptospirosis.
Project description:This SuperSeries is composed of the following subset Series: GSE30721: Profiling proteome-scale antibody responses to M. tuberculosis proteins in sera of macaques infected with M. tuberculosis GSE30722: Profiling proteome-scale antibody responses to M. tuberculosis proteins in TB suspect's sera Refer to individual Series
Project description:Systemic lupus erythematosus is progressive, immune complex-mediated autoimmune disease targeting numerous organs. A central feature of the disease is the development of antibodies against nuclear components, including DNA. Antibodies against double-stranded DNA are so characteristic of this disease that their detection constitutes one of the criteria for diagnosis. We examined the formation of immune complexes on the surface of autoantigen microarrays incubated in the sera of 39 inactive and 22 active lupus patients and of 31 control subjects. Three different kinds of nucleic acids, dsDNA, ssDNA and RNA were used as antigens, along with chromatin (nucleosomal extract) and several other reference molecules. The composition with respect to IgG, IgM and complement components C3 and C4 was determined. We find that while IgM and C4 are physiological components of immune complexes formed from nucleic acids, both IgG and C3 are extremely characteristic of lupus patients. Complement C4 deposition changes were not consistent: these increased on ssDNA and RNA, decreased on chromatin and did not change significantly on dsDNA. The presence of IgG and C3 in the immune complexes formed from different nucleic acids was characteristic for both active and inactive lupus patients. Receiver-operating curve statistics indicate that C3 deposition measurements can improve the efficiency of identification of inactive lupus patients. These observations reveal the complexity of immune profile changes accompanying SLE. C3, IgM, C4 and IgG binding in 92 human serum samples were examined using custom-made protein arrays
Project description:Enterovirus 71 (EV71) and coxsackievirus A16 (CA16) are major causative agents for hand, foot and mouth disease (HFMD). In China, more than 70,756 children were infected and 40 died from the disease in recent years. This study aimed to develop a protein chip that can simultaneously detect and differentiate the antibodies induced by EV71 and CA16 simultaneously. The structure protein vp1 and vp3 from the two viruses were purified and spotted onto an aldehyde groupmodified glass slide at 1mg/ml. After that, the protein chip was reacted with the corresponding positive serum against these viruses, hybridized with a Cy3-labeled secondary antibody and scanned using the popular GenePix 4000B Microarray Scanner. In this study, Both IgG and IgM serum antibody to EV71 and CA16 were detected using protein Chip. The results showed that this method could hybridize specifically with the corresponding antibodies with strong signals and without cross-hybridization. The data also confirmed the proposed method's specificity, sensitivity, and convenience. In conclusion, this protein chip can be used to differentiate the antibodies induced by the EV71and CA16.