Technical Development Project 1: Measuring the Immunome: Genomic Approaches to B Cell Repertoire. In this project, we propose to develop new technological applications of next generation DNA sequencing, high throughput PCR chips, and microfluidic single cell processors to perform systemic analyses of the human B cell repertoire. Using B cells purified from human peripheral blood, we will apply these technologies to try to understand the effects of three phenomena on immune repertoire: aging, genetic background and vaccination history. Four groups of subjects will be studied: 1. age groups - children, young adults and elderly;2. genetic background - identical twins vs fraternal twins vs not-related subjects;3. vaccination history - pre, post and multiple immunizations;4. vaccination type - the trivalent inactivated influenza vaccine (TIV) and the live attenuated influenza vaccine (LAIV). This study will provide the first comprehensive and systematic measurements of B cell repertoire at three different anatomic levels, V/D/J/C exon usage, immunoglobulin (Ig) gene deep sequencing, and single cell gene expression analysis.
Our specific aims are:
Aim 1. Develop a quick and quantitative assay to measure the V, D J, and C exon usage landscape.
Aim 2. Develop a high throughput sequencing protocol to perform deep sequencing of Ig heavy chain gene diversity.
Aim 3. Develop a microfluidic parallel processor for single cell analysis of B cell repertoire.
Aim 4. Dissect the influences of aging, genetic background, and vaccination history on the human B cell repertoire.

Public Health Relevance

Data generated from this study will advance our understanding of the B cell repertoire in response to influenza vaccine, which in turn will increase our knowledge of how influenza interacts with the immune system. In addition, technologies developed here may prove useful to advance clinical diagnostics for influenza and other pathogens.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program--Cooperative Agreements (U19)
Project #
5U19AI057229-07
Application #
8060521
Study Section
Special Emphasis Panel (ZAI1)
Project Start
Project End
Budget Start
2010-04-01
Budget End
2011-03-31
Support Year
7
Fiscal Year
2010
Total Cost
$262,773
Indirect Cost
Name
Stanford University
Department
Type
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94305
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