Sequencing the Peripheral Blood B and T cell Repertoire - Quantifying robustness and limitations

Here we establish experimental and analytical protocols for quantifying from the peripheral blood compartment of healthy human individuals the immune repertoire for the Complementarity determining region 3 (CDR3) of the variable regions of TCRbeta (CDRβ3) and the IgG heavy chain (CDRH1, CDRH2, CDRH3). The results demonstrate that 40 ml of blood are sufficient to reliably capture the 10,000 most common TCRbeta and 1000 most common IgG and determine their relative frequency in the circulation. We conclude that by using an accessible sample size of human PBMC one is able to robustly monitor alterations in the immune repertoire.