Human Peripheral Blood Antibodies with Long HCDR3s Are Established Primarily at Original Recombination Using a Limited Subset of Germline Genes
Figure 3
Long HCDR3s correlate with N-addition, P-addition and germline gene usage.
For all figure sections, the leftmost panel corresponds to peripheral blood antibody sequences from Group 1 (n = 4) and Group 2 (n = 3) healthy donors. The middle panel corresponds to antibody sequences from Group 1 donors, segregated by B cell subset. The rightmost panel corresponds to peripheral blood antibody sequences from HIV-infected donors (n = 4). (A) Peripheral blood antibody sequences were grouped by HCDR3 length (in amino acids) and the average N-addition length and P-addition length (both in nucleotides) was calculated for each HCDR3 length group. The mean length ± SEM is shown. Regression analysis of N-addition length produced a non-linear, exponential curve of best fit. Regression analysis of P-addition length produced a linear best fit. (B) Peripheral blood antibody sequences were grouped by HCDR3 length and the frequency of sequences encoding either diversity gene family 2 (D2) or diversity gene family 3 (D3) was calculated for each HCDR3 length group. The mean frequency of D2/D3 gene family use ± SEM for each HCDR3 length group is shown. Regression analysis produced a non-linear, sigmoidal curve of best fit. (C) Peripheral blood antibody sequences were grouped by HCDR3 length and the frequency of sequences encoding joining gene 6 (JH6) was calculated for each HCDR3 length group. The mean frequency of D2/D3 gene family use ± SEM for each HCDR3 length group is shown. Regression analysis produced a non-linear, sigmoidal curve of best fit. (D) The frequency of sequences encoding both the JH6 germline gene and D2/D3 germline gene family members was determined for each HCDR3 length group. The mean frequency of JH6/D2/D3 gene family use ± SEM for each HCDR3 length group is shown. Non-linear regression analysis produced a sigmoidal curve of best fit.