High-throughput sequencing of human immunoglobulin variable regions with subtype identification
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Autor:innen
Schanz, Merle
Liechti, Thomas
Zagordi, Osvaldo
Reddy, Sai T.
Günthard, Huldrych F.
Trkola, Alexandra
Huber, Michael
Autor:in (Körperschaft)
Publikationsdatum
03.11.2014
Typ der Arbeit
Studiengang
Typ
01A - Beitrag in wissenschaftlicher Zeitschrift
Herausgeber:innen
Herausgeber:in (Körperschaft)
Betreuer:in
Übergeordnetes Werk
PLOS ONE
Themenheft
DOI der Originalpublikation
Link
Reihe / Serie
Reihennummer
Jahrgang / Band
9
Ausgabe / Nummer
11
Seiten / Dauer
Patentnummer
Verlag / Herausgebende Institution
Public Library of Science
Verlagsort / Veranstaltungsort
Auflage
Version
Programmiersprache
Abtretungsempfänger:in
Praxispartner:in/Auftraggeber:in
Zusammenfassung
The humoral immune response plays a critical role in controlling infection, and the rapid adaptation to a broad range of pathogens depends on a highly diverse antibody repertoire. The advent of high-throughput sequencing technologies in the past decade has enabled insights into this immense diversity. However, not only the variable, but also the constant region of antibodies determines their in vivo activity. Antibody isotypes differ in effector functions and are thought to play a defining role in elicitation of immune responses, both in natural infection and in vaccination. We have developed an Illumina MiSeq high-throughput sequencing protocol that allows determination of the human IgG subtype alongside sequencing full-length antibody variable heavy chain regions. We thereby took advantage of the Illumina procedure containing two additional short reads as identifiers. By performing paired-end sequencing of the variable regions and customizing one of the identifier sequences to distinguish IgG subtypes, IgG transcripts with linked information of variable regions and IgG subtype can be retrieved. We applied our new method to the analysis of the IgG variable region repertoire from PBMC of an HIV-1 infected individual confirmed to have serum antibody reactivity to the Membrane Proximal External Region (MPER) of gp41. We found that IgG3 subtype frequencies in the memory B cell compartment increased after halted treatment and coincided with increased plasma antibody reactivity against the MPER domain. The sequencing strategy we developed is not restricted to analysis of IgG. It can be adopted for any Ig subtyping and beyond that for any research question where phasing of distant regions on the same amplicon is needed.
Schlagwörter
Fachgebiet (DDC)
600 - Technik, Medizin, angewandte Wissenschaften
Veranstaltung
Startdatum der Ausstellung
Enddatum der Ausstellung
Startdatum der Konferenz
Enddatum der Konferenz
Datum der letzten Prüfung
ISBN
ISSN
1932-6203
Sprache
Englisch
Während FHNW Zugehörigkeit erstellt
Nein
Publikationsstatus
Veröffentlicht
Begutachtung
Peer-Review der ganzen Publikation
Open Access-Status
Gold
Lizenz
Zitation
SCHANZ, Merle, Thomas LIECHTI, Osvaldo ZAGORDI, Enkelejda MIHO, Sai T. REDDY, Huldrych F. GÜNTHARD, Alexandra TRKOLA und Michael HUBER, 2014. High-throughput sequencing of human immunoglobulin variable regions with subtype identification. Shan LU (Hrsg.), PLOS ONE. 3 November 2014. Bd. 9, Nr. 11. DOI 10.1371/journal.pone.0111726. Verfügbar unter: https://doi.org/10.26041/fhnw-9951