The proposed studies build on our previous findings indicating that interactive T- and B-like lymphocytes are basic features of the adaptive immune system in both jawless and jawed vertebrates, although jawless vertebrates (lampreys and hagfish) generate variable lymphocyte receptors (VLR) for antigen recognition by recombinatorial conversion of incomplete germline VLR genes (VLRA, VLRA and VLRC) into fully assembled VLR genes using diverse leucine-rich repeat (LRR) donor sequences. Recent studies indicate that while VLRA gene assembly coincides with expression of cytidine deaminase 1 (CDA1) within the lymphoepithelial thymoid gill region, coincident VLRB and CDA2 expression instead occurs primarily in hematopoietic tissues. A comprehensive analysis is proposed to elucidate the development, distribution and function of the B-like VLRB lymphocytes in lampreys. The overall goals are to better understand B cell biology in a jawless vertebrate, in particular the mechanism of generating VLRB antibody diversity, and to harness the lamprey antibody system for biomedical purposes. An efficient newly-developed scheme for lamprey immunization will be coupled with subsequent high-throughput screening of VLRB cDNA libraries from immunized animals to obtain specific monoclonal VLRB antibodies with high binding affinity for model antigens of biomedical significance. The first specific aim is to elucidate the sequence diversity and possibility of VLRB antibody affinity maturation during lamprey B cell responses to primary and booster immunizations with model protein antigens. The second specific aim is to determine the lamprey CDA2 structure, functional potential, and expression as a function of the differentiation status and tissue localization of VLRB lineage cells. The third specific aim is to produce recombinant VLRB antibodies with novel specificities for use as diagnostic reagents for medically important infectious agents and tumor cell types. These interrelated studies will provide insight into the evolution of the alternative B cell types f jawless and jawed vertebrates and will also yield novel monoclonal lamprey antibodies with specificity for infectious agents and tumor cell antigens of humans for use as diagnostic and disease-monitoring reagents.

Public Health Relevance

The proposed studies will yield insight into the basic principles of B cell evolution in vertebrates. They will elucidate the mechanism(s) for diversification of the variable lymphocyte receptor (VLR) repertoire in lampreys and provide unique VLR antibodies as novel reagents for use in identifying infectious agents and cancer cells in humans.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI072435-07
Application #
8297730
Study Section
Cellular and Molecular Immunology - B Study Section (CMIB)
Program Officer
Leitner, Wolfgang W
Project Start
2007-01-15
Project End
2017-01-31
Budget Start
2012-02-01
Budget End
2013-01-31
Support Year
7
Fiscal Year
2012
Total Cost
$387,500
Indirect Cost
$137,500
Name
Emory University
Department
Pathology
Type
Schools of Medicine
DUNS #
066469933
City
Atlanta
State
GA
Country
United States
Zip Code
30322
Das, Sabyasachi; Sutoh, Yoichi; Hirano, Masayuki et al. (2016) Characterization of Lamprey BAFF-like Gene: Evolutionary Implications. J Immunol 197:2695-703
Yu, Cuiling; Liu, Yanling; Chan, Justin Tze Ho et al. (2016) Identification of human plasma cells with a lamprey monoclonal antibody. JCI Insight 1:
Das, Sabyasachi; Li, Jianxu; Hirano, Masayuki et al. (2015) Evolution of two prototypic T cell lineages. Cell Immunol 296:87-94
Tillotson, Benjamin J; Lajoie, Jason M; Shusta, Eric V (2015) Yeast Display-Based Antibody Affinity Maturation Using Detergent-Solubilized Cell Lysates. Methods Mol Biol 1319:65-78
Han, Qifeng; Das, Sabyasachi; Hirano, Masayuki et al. (2015) Characterization of Lamprey IL-17 Family Members and Their Receptors. J Immunol 195:5440-51
Altman, Meghan O; Bennink, Jack R; Yewdell, Jonathan W et al. (2015) Lamprey VLRB response to influenza virus supports universal rules of immunogenicity and antigenicity. Elife 4:
Das, Sabyasachi; Li, Jianxu; Holland, Stephen J et al. (2014) Genomic donor cassette sharing during VLRA and VLRC assembly in jawless vertebrates. Proc Natl Acad Sci U S A 111:14828-33
Holland, Stephen J; Gao, Mingming; Hirano, Masayuki et al. (2014) Selection of the lamprey VLRC antigen receptor repertoire. Proc Natl Acad Sci U S A 111:14834-9
Nakahara, Hirotomo; Herrin, Brantley R; Alder, Matthew N et al. (2013) Chronic lymphocytic leukemia monitoring with a Lamprey idiotope-specific antibody. Cancer Immunol Res 1:223-8
Hirano, Masayuki; Guo, Peng; McCurley, Nathanael et al. (2013) Evolutionary implications of a third lymphocyte lineage in lampreys. Nature 501:435-8

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