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

Natalia T Freund; Johannes F Scheid; Hugo Mouquet; Michel C Nussenzweig

doi:10.1016/j.jim.2015.01.011

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

J Immunol Methods. 2015 Mar:418:61-5. doi: 10.1016/j.jim.2015.01.011. Epub 2015 Feb 7.

Authors

Natalia T Freund¹, Johannes F Scheid¹, Hugo Mouquet², Michel C Nussenzweig³

Affiliations

¹ Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 10065, USA.
² Laboratory of Humoral Response to Pathogens, Department of Immunology, Institut Pasteur, Paris 75015, France.
³ Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 10065, USA; Howard Hughes Medical Institute, The Rockefeller University, New York, NY 10065, USA. Electronic address: [email protected].

PMID: 25667013
DOI: 10.1016/j.jim.2015.01.011

Abstract

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.

Keywords: Anti-HIV-1 antibodies; Human Ig lambda; PCR; Single cell sorting; Somatic hypermutation.

MeSH terms

Flow Cytometry
HIV Infections / genetics*
HIV Infections / immunology*
Humans
Immunoglobulin lambda-Chains / genetics*
Immunoglobulin lambda-Chains / immunology*
Leukocytes, Mononuclear / immunology
Leukocytes, Mononuclear / virology
Mutagenesis / genetics*
Mutation / genetics*

Substances

Immunoglobulin lambda-Chains