This came about upon realizing that patients with lymphoproliferative disorders could be considered the human equivalent of IG transgenic mice, since both expressed almost exclusively a single B lymphocyte with a single IGHV-IGHD-IGHJ (IGHV-D-J) rearrangement
This came about upon realizing that patients with lymphoproliferative disorders could be considered the human equivalent of IG transgenic mice, since both expressed almost exclusively a single B lymphocyte with a single IGHV-IGHD-IGHJ (IGHV-D-J) rearrangement. minimizes side effects. Historical context The story from Southampton (UK) The Tenovus Institute was established in 1970 at the new medical school in Southampton and George and I moved there from Oxford. With the help of Tenovus funding, I established a laboratory aimed at studying B cells, while George focused on antibody therapy. The new tools of immunogenetics became available Rabbit Polyclonal to ATP1alpha1 in the early 1990s, and my lab seized on the new opportunity to understand immunoglobulins. Part of the drive arose from our studies of cold agglutinin (CA) disease, where Myf Spellerberg’s precious CA-secreting single B cells had been handed to other laboratories for the then novel approach of DNA sequencing. Those labs had the fun of showing that they were all derived from the IGHV4C34 gene. We immediately bought a PCR machine and Caroline Chapman and I set up a molecular biology laboratory and taught ourselves how to sequence IGV genes, initially using gel-based methods. Protein sequencing had already been applied to clonal Igs secreted by plasma cells, but DNA sequencing added crucial information on V(D)J recombination, somatic hypermutation and isotype switch. Interpretation of sequence data however totally relied on having available the libraries of human IGV, D and J genes existing in the unrearranged DNA, which were provided by the labs of Tasuku Honjo, Fred Alt and Greg Winter. For once, human genetics was ahead of the mouse, and we could map the clonal history of any B cell. For normal B cells, the range and level of IGV gene usage was investigated by Peter Lipsky’s group.3 Strangely the functional repertoire in circulating B cells did not simply reflect the available library and a similar selection was evident in different ethnic groups. Rearrangements of one, or sometimes both, alleles were mapped giving information on non-functional IGHV genes. Leaning around the revelations of somatic hypermutation from Cesar Milstein, the Irosustat Lipsky lab analyzed mutational patterns and revealed differential distribution across IGV sequences. In what seems to be an evolutionary mechanism, most hot spots are in the complementarity-determining regions.4 In a fairly short time we had sequenced B cell IGHV genes in every possible situation ranging from normal B cells, IgE, EBV contamination, autoimmunity to the full range Irosustat of B-cell tumors. Comparison with the databases Irosustat allowed insights into the point of differentiation reached by the transformed B cell, and any subsequent changes. During this time, Terry Hamblin, based in the then non-university hospital at Bournemouth, provided the important bridge from his common patient clinic to Irosustat our lab. One day we discussed whether we should look in detail at the most common B-cell tumor, chronic lymphocytic leukemia (CLL). I had formed already had a quick look at cases from David Oscier who was focused on chromosomal abnormalities, and we showed that cases with trisomy 12 tended to have unmutated IGHV genes whereas those with a single 13q14 abnormality had higher mutational levels.5 This confirmed the view from others that CLL was heterogeneous, but we had only small numbers. Terry and I decided to do a blitz of VH sequencing around the large number of clinically well-documented cases he and David had stored. It meant directing the lab effort toward this, with Zadie Davis, then in my lab, at the forefront. With RNA as the preferred source to avoid nonfunctional IGHVs,.