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Blood, Vol. 111, Issue 2, 741-749, January 15, 2008
Increased survival is a selective feature of human circulating antigen-induced plasma cells synthesizing high-affinity antibodies
Blood González-García et al.
111: 741
Supplemental material for: Gonzalez-Garcia et al
Files in this Data Supplement:
- Table S1. Number of analyzed IGVH3 sequences per donor and PC subset (PDF, 13.1 KB).
- Figure S1. Distributions of mutations (PDF, 71.2 KB). -
Distribution of total mutations (left panel) and total substituted amino acids (right panel) observed in the IGVH3 gene sequenced from purified tetCNEG PC (black bars), tetC INT PC (open bars) and tetCHIGH PC (grey bars). The results are expressed as the mean ± s.e.m. of 33, 32 and 16 sequences, for tetC NEG PC, tetC INT PC and tetC HIGH PC, respectively. Asterisks indicate results that were significantly different by the Studentīs T test (p<0.04).
- Figure S2. Four rooted phylogenetic trees from alignment of IGVH3 sequences (PDF, 372 KB). -
Four rooted phylogenetic trees from alignment of IGVH3 sequences were obtained from each experiment (donor) using SDSC Biology Workbench 3.2. Clonally related and un-related sequences were established according to their either unique or shared VH-DH-JH rearrangement, as previously reported. (Schittek, B. and K. Rajewsky.1992. Natural Occurrence and Origin of Somatically Mutated Memory B Cells in Mice, J Exp Med 176:427-438; Jacob, J. and G. Kelsoe. 1992. In Situ Studies of the Primary Immune Response to (4-hydroxy-3-nitrophenyl)acetyl. II. A Common Clonal Origin for Periarteriolar Lymphoid Sheath-associated Foci and Germinal Centers, J Exp Med 176:679-687). Genealogical relationships of IGVH3 sequences are indicated within each circle; these sequences come from a common progenitor and were identified by the shared CDR3 sequence. Each isolated sequence is named with the PC subset type followed by the corresponding GenBank accession number.
- Figure S3. Comparison of the clonally-related sequences obtained from experiments 1, 2, 3 and 4 (PDF, 32.0 KB). -
The germ-line IGVH3 gene in each case is shown, and identity with the germ-line sequence is denoted by dashes. Mutations are indicated in each case. CDR1, CDR2 and CDR3 regions are indicated in bold. The CDR3 sequence is shown for the first sequence of the families and identity with this sequence is indicated for the rest of the associated sequences as before. The corresponding lineage trees are shown in Fig. 4S. tetC HIGH and tetC INT PC sequences contain multiple shared mutations, indicating clonal diversification.
- Figure S4. Genealogical relationship of the VDJ sequences obtained from experiment 1 (A), experiment 2 (B), experiment 3 (C and D) and experiment 4 (E and F) (PDF, 58.8 KB). -
The solid circles indicate observed mutated sequences (cells) with the PC subset and the GenBank accession number inside. Circles of broken lines denote hypothetical intermediate sequences (cells) that were not found but have mutations that are shared by all subsequent sequences (cells). Precursor lymphocyte (P.L.) identifies the germline of the corresponding VH3 sub-family: VH3-23 (A and B), VH3-30 (C), VH3-9 (D), VH3-11 (E) and VH3-21 (F). Numbers in branches indicate the number of mutations to the next node.
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