A multidisciplinary three-pillar approach was employed.
The first referred to the extensive immunogenetic analysis using high throughput sequencing techniques (next generation sequencing, NGS) that unveiled the imprints of the interaction of BcR IGs with selecting antigens. The second concerned the investigation of the actual identity of the antigenic elements involved in the selection and activation of (pre)malignant B cell clones, by expressing the BcR IG as soluble molecules through recombinant DNA technology and performing binding assays against a broad range of antigenic targets, including self and foreign antigens. Collectively, these studies offered strong evidence for functional pressure by specific antigenic elements. In addition, our novel findings emphasized the importance of BcR IG isotype in modulating fine antigen specificity through the generation of a large library of soluble CLL IGs of the ‘authentic’ isotype (same as that expressed by the leukemic clone): by using these molecules in antigen binding assays we confirmed and further extended the existence of a tight link between self- and poly-reactive patterns of BcR IG and aggressive disease in CLL, in support of a scenario where abundancy of antigenic stimuli provide a selection advantage to B cell clones. Last but emphatically not least, within IGpath we examined by NGS the BcR IG repertoire of a large number of MBL cases and, for the first time, the MBL BcR IG in terms of antigenic reactivity properties, hence, contributing into the comprehension of the BcR IG-antigen interaction processes leading to monoclonal cell expansions that could eventually drive neoplastic transformation. In addition, we completed the largest to-date functional analysis of SMZL BcR IG and showed intriguing features of BcR IG-antigen patterns of interaction in the disease.
In the third pillar we explored the role of BcR homotypic interactions in shaping the clonal behavior in a large cohort of CLL cases. Using appropriate biochemical technologies, we assessed the magnitude of BcR self-association and significantly extended our previous observations unveiling links between the qualities of the BcR self-association and the disease outcome.
The main results of IGpath have been presented at the 60th and 61st Annual Meetings of the American Society of Hematology, the prime scientific event in hematology. At the national level, IGpath results have been presented at the 30th Annual Congress of Hellenic Society of Hematology. In addition, the progress of the project has been systematically communicated through IGpath’s website.
