The human preBCR uses CDR-H3 amino acid composition to control early B cell selection and prevent oncogenesis

Abstract

Abstract Human B cell repertoire selection depends on the composition of the complementary determining heavy chain (CDR-H3), which lies at the center of the antigen binding site. During B cell development, preB cells express a preB cell receptor (preBCR), which is composed of μH chain bound to surrogate light chain (SLC). In mouse, the VpreB R 101D 57R 51motif of the SLC selects against μH chains with hydrophobic CDR-H3s. Abnormal selection at the preBCR stage predisposes to autoreactive B cell survival and leukemia. We sought to study preBCR selection in healthy human individuals versus acute lymphoblastic leukemia (ALL) patients to dissect the mechanism of formation of the human protective CDR-H3 repertoire. We hypothesized that expression of a preBCR with an abnormal amino acid content prevents differentiation of the ALL blasts into a late preB and then a mature B cell, promoting oncogenic survival. We sorted early B cells from bone marrow swabs of healthy controls and ALL patients. We extracted RNA, deep sequenced preB cells and leukemic blasts and then analyzed their CDR-H3 based on amino acid content using bioinformatics and structure modeling. Our results suggest that in healthy individuals, similar to mouse, the human preBCR selects for CDR-H3 based on amino acid content with a preference for tyrosine (Y) and negative selection of hydrophobic antigen binding sites. The CDR-H3s of ALL leukemic blasts have amino acid characteristics similar to the negatively selected antigen binding sites of healthy individuals. In conclusion, this type of analysis might be used to monitor optimal immunoglobulin repertoire diversification. R21AI163555