Amplification of highly mutated human Ig lambda light chains from an HIV-1 infected patient.

Isolation and characterization of anti HIV-1 broadly neutralizing antibodies (bNAbs) have elucidated new epitopes and sites of viral vulnerability. Anti-HIV-1 bNAbs typically show high levels of somatic mutations in their variable region genes. This feature potentially limits antibody identification, since the mutated antibody sequences are no longer complimentary to primers designed based on germline antibody sequences. Here we report a new set of primers for Igλ light chains that aligns to the 5' end of the leader sequence and is highly efficient for the amplification of antibodies that contain mutations and deletions in the 5' end of human Igλ.

[1]  M. Nussenzweig,et al.  AID targeting in antibody diversity. , 2011, Advances in immunology.

[2]  Florian Klein,et al.  Antibodies in HIV-1 Vaccine Development and Therapy , 2013, Science.

[3]  Tongqing Zhou,et al.  Structure-Based Stabilization of HIV-1 gp120 Enhances Humoral Immune Responses to the Induced Co-Receptor Binding Site , 2009, PLoS pathogens.

[4]  M. Nussenzweig,et al.  Broadly Neutralizing Antibodies and Viral Inducers Decrease Rebound from HIV-1 Latent Reservoirs in Humanized Mice , 2014, Cell.

[5]  Ron Diskin,et al.  Sequence and Structural Convergence of Broad and Potent HIV Antibodies That Mimic CD4 Binding , 2011, Science.

[6]  F. Pereyra,et al.  Persistent low-level viremia in HIV-1 elite controllers and relationship to immunologic parameters. , 2009, The Journal of infectious diseases.

[7]  J. Mascola,et al.  Passive transfer of modest titers of potent and broadly neutralizing anti-HIV monoclonal antibodies block SHIV infection in macaques , 2014, The Journal of experimental medicine.

[8]  Tongqing Zhou,et al.  Somatic Mutations of the Immunoglobulin Framework Are Generally Required for Broad and Potent HIV-1 Neutralization , 2013, Cell.

[9]  M. Nussenzweig,et al.  Predominant Autoantibody Production by Early Human B Cell Precursors , 2003, Science.

[10]  Michel C Nussenzweig,et al.  Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning. , 2008, Journal of immunological methods.

[11]  Martin A. Nowak,et al.  Antibody neutralization and escape by HIV-1 , 2003, Nature.

[12]  Florian Klein,et al.  Structural Insights on the Role of Antibodies in HIV-1 Vaccine and Therapy , 2014, Cell.

[13]  Ron Diskin,et al.  HIV therapy by a combination of broadly neutralizing antibodies in humanized mice , 2012, Nature.

[14]  Ron Diskin,et al.  Structural basis for germ-line gene usage of a potent class of antibodies targeting the CD4-binding site of HIV-1 gp120 , 2012, Proceedings of the National Academy of Sciences.

[15]  J. Mascola,et al.  Most rhesus macaques infected with the CCR5-tropic SHIVAD8 generate cross-reactive antibodies that neutralize multiple HIV-1 strains , 2012, Proceedings of the National Academy of Sciences.

[16]  F. Pereyra,et al.  A method for identification of HIV gp140 binding memory B cells in human blood. , 2009, Journal of immunological methods.

[17]  M. Nussenzweig,et al.  Memory B Cell Antibodies to HIV-1 gp140 Cloned from Individuals Infected with Clade A and B Viruses , 2011, PloS one.

[18]  M. Nussenzweig,et al.  Broadly Neutralizing Anti-HIV-1 Antibodies Require Fc Effector Functions for In Vivo Activity , 2014, Cell.

[19]  Maxim N. Artyomov,et al.  Polyreactivity increases the apparent affinity of anti-HIV antibodies by heteroligation , 2010, Nature.

[20]  C. Milstein,et al.  Mutation Drift and Repertoire Shift in the Maturation of the Immune Response , 1987, Immunological reviews.

[21]  C. Milstein,et al.  The Dynamic Nature of the Antibody Repertoire , 1988, Immunological reviews.

[22]  Florian Klein,et al.  Computational analysis of anti–HIV-1 antibody neutralization panel data to identify potential functional epitope residues , 2013, Proceedings of the National Academy of Sciences.

[23]  Richard T. Wyatt,et al.  Broad diversity of neutralizing antibodies isolated from memory B cells in HIV-infected individuals , 2009, Nature.

[24]  M. Nussenzweig,et al.  Autoreactivity in human IgG+ memory B cells. , 2007, Immunity.