Project description:Proteomic identification and characterization of antibodies comprising the serological response to antigen can provide unique insight into the functional dynamics of adaptive immunity. We have developed a novel method to overcome the technical challenges which previously limited the direct analysis of immunoglobulin proteins in serum, as demonstrated by the identification of human anti-tetanus toxoid (TT) immunoglobulin G (IgG) proteins following booster vaccination. We analyzed the serum IgG repertoire across four time-points corresponding to pre-vaccination, 7 days, 3 months, and 9 months post vaccination. Antigen-specific antibodies were affinity purified against immobilized TT protein and sequenced by bottom-up nanoLC-MS/MS. Interpretation of mass spectra required a custom reference database of IgG heavy and light chain variable sequences determined by NextGen RNA sequencing of the donor's circulating plasmablasts and memory B cells following booster vaccination.
Project description:Rice (Oryza sativa L.) is a candidate crop for production of plant-based vaccines by genetic engineering technologies. MucoRice-CTB has been developed as a rice-seed-based vaccine against cholera by transgenic expression of modified cholera toxin B-subunit (CTB) under the RNA interference (RNAi)-mediated suppression of endogenous seed storage proteins. Here, we performed non-targeted metabolomic profiling of MucoRice-CTB to understand the overall effects of the genetic engineering on rice seed metabolism using gas chromatography/time-of-flight mass spectrometry.
Project description:1. Charge-shift electrophoresis showed that cholera toxin and its subunits have no hydrophobic surfaces. 2. Amino-acid composition and sequence data suggested that the proteins have no masked hydrophobic regions. 3. The A subunit of cholera toxin may interact with polar molecules in the membrane to exert its effect inside the cell. 4. The only hydrophobic part of tetanus toxin was the H-chain.
Project description:Diphtheria, tetanus, and pertussis (DTP) are infectious diseases caused by toxin-producing bacteria that can be fatal, especially in children. Despite the availability of combined DTP vaccines for more than 60 years, a comprehensive understanding of how human antibodies respond to DTP vaccination, which helps further improve the vaccine remains elusive. Here, we aimed to characterize toxin-specific antibody repertoires following DTP vaccination. To this end, CD19+CD27+ antigen-experienced B cells obtained from four healthy donors 7-days post vaccination were sorted for toxin-binding and non-toxin-binding B cells, using each of the DTP recombinant toxins (DT, TT, and PT) as fluorescent bait. 10X single-cell immune profiling was then performed, followed by in silico antibody sequence clustering.
Project description:Bovine anaplasmosis is an arthropod-borne hemolytic disease caused by Anaplasma marginale. While only a few Anaplasma marginale strains have been reported, no Mexican strains have been reported. Due to the genetic diversity of A. marginale, the genome of the strain Mex-01-001-01, isolated in Mexico, represents a new source of information.
Project description:We observed the expression profile of the total mRNA of crp (TTHA1437) deletion mutant of Thermus thermophilus HB8 strain grown in a rich (TT) medium at 70 degC
Project description:Mechanisms of poor responses to vaccines remain unknown. Hepatitis B virus-naïve elderly subjects received three vaccines, including a vaccine against hepatitis B virus (HBV). Pre-vaccination high dimensional analyses of blood using transcriptional profiling and flow cytometry revealed that subjects having increased memory B cell frequencies and higher expression of genes downstream of B cell receptor signaling responded more strongly to the HBV vaccine whereas subjects having higher expression of inflammatory related genes and greater numbers of activated innate immune cells showed a weaker response to this vaccine. The heme-induced response was associated with the poor response to the hepatitis B vaccine. Transcriptional profiling and flow cytometry results were validated in a distinct set of elderly subjects with accuracy greater than 60%. Our study is the first that identifies baseline predictors of responses to vaccines in a population of subjects known to be highly susceptible to infections.
