Absence of anti-hypocretin receptor 2 autoantibodies in post pandemrix narcolepsy cases

Guo Luo, Ling Lin, Louis Jacob, Mélodie Bonvalet, Aditya Ambati, Giuseppe Plazzi, Fabio Pizza, Ryan Leib, Christopher M. Adams, Markku Partinen, Emmanuel Jean Marie Mignot

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Background: A recent publication suggested molecular mimicry of a nucleoprotein (NP) sequence from A/Puerto Rico/8/1934 (PR8) strain, the backbone used in the construction of the reassortant strain X-179A that was used in Pandemrix® vaccine, and reported on anti-hypocretin (HCRT) receptor 2 (anti-HCRTR2) autoantibodies in narcolepsy, mostly in post Pandemrix® narcolepsy cases (17 of 20 sera). In this study, we re-examined this hypothesis through mass spectrometry (MS) characterization of Pandemrix®, and two other pandemic H1N1 (pH1N1)-2009 vaccines, Arepanrix® and Focetria®, and analyzed anti-HCRTR2 autoantibodies in narcolepsy patients and controls using three independent strategies. Methods: MS characterization of Pandemrix® (2 batches), Arepanrix® (4 batches) and Focetria® (1 batch) was conducted with mapping of NP 116I or 116M spectrogram. Two sets of narcolepsy cases and controls were used: 40 post Pandemrix® narcolepsy (PP-N) cases and 18 age-matched post Pandemrix® controls (PP-C), and 48 recent (6 months) early onset narcolepsy (EO-N) cases and 70 age-matched other controls (O-C). Anti-HCRTR2 autoantibodies were detected using three strategies: (1) Human embryonic kidney (HEK) 293T cells with transient expression of HCRTR2 were stained with human sera and then analyzed by flow cytometer; (2) In vitro translation of [35S]-radiolabelled HCRTR2 was incubated with human sera and immune complexes of autoantibody and [35S]-radiolabelled HCRTR2 were quantified using a radioligand-binding assay; (3) Optical density (OD) at 450 nm (OD450) of human serum immunoglobulin G (IgG) binding to HCRTR2 stably expressed in Chinese hamster ovary (CHO)-K1 cell line was measured using an in-cell enzyme-linked immunosor-bent assay (ELISA). Results: NP 116M mutations were predominantly present in all batches of Pandemrix®, Arepanrix® and Focetria®. The wild-type NP109-123 (ILYDKEEIRRIWRQA), a mimic to HCRTR234-45 (YDDEEFLRYLWR), was not found to bind to DQ0602. Three or four subjects were found positive for anti-HCRTR2 autoantibodies using two strategies or the third one, respectively. None of the post Pandemrix® narcolepsy cases (0 of 40 sera) was found positive with all three strategies. Conclusion: Anti-HCRTR2 autoantibody is not a significant biological feature of narcolepsy or of post Pandemrix® autoimmune responses.

Original languageEnglish
Article numbere0187305
JournalPLoS One
Volume12
Issue number12
DOIs
Publication statusPublished - Dec 1 2017

Fingerprint

Orexin Receptors
Narcolepsy
autoantibodies
Autoantibodies
receptors
nucleoproteins
Nucleoproteins
Serum
Mass spectrometry
Assays
Mass Spectrometry
mass spectrometry
molecular mimicry
Vaccines
vaccines
antigen-antibody complex
Molecular Mimicry
autoimmunity
pandemrix
narcolepsy

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Luo, G., Lin, L., Jacob, L., Bonvalet, M., Ambati, A., Plazzi, G., ... Mignot, E. J. M. (2017). Absence of anti-hypocretin receptor 2 autoantibodies in post pandemrix narcolepsy cases. PLoS One, 12(12), [e0187305]. https://doi.org/10.1371/journal.pone.0187305

Absence of anti-hypocretin receptor 2 autoantibodies in post pandemrix narcolepsy cases. / Luo, Guo; Lin, Ling; Jacob, Louis; Bonvalet, Mélodie; Ambati, Aditya; Plazzi, Giuseppe; Pizza, Fabio; Leib, Ryan; Adams, Christopher M.; Partinen, Markku; Mignot, Emmanuel Jean Marie.

In: PLoS One, Vol. 12, No. 12, e0187305, 01.12.2017.

Research output: Contribution to journalArticle

Luo, G, Lin, L, Jacob, L, Bonvalet, M, Ambati, A, Plazzi, G, Pizza, F, Leib, R, Adams, CM, Partinen, M & Mignot, EJM 2017, 'Absence of anti-hypocretin receptor 2 autoantibodies in post pandemrix narcolepsy cases', PLoS One, vol. 12, no. 12, e0187305. https://doi.org/10.1371/journal.pone.0187305
Luo, Guo ; Lin, Ling ; Jacob, Louis ; Bonvalet, Mélodie ; Ambati, Aditya ; Plazzi, Giuseppe ; Pizza, Fabio ; Leib, Ryan ; Adams, Christopher M. ; Partinen, Markku ; Mignot, Emmanuel Jean Marie. / Absence of anti-hypocretin receptor 2 autoantibodies in post pandemrix narcolepsy cases. In: PLoS One. 2017 ; Vol. 12, No. 12.
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abstract = "Background: A recent publication suggested molecular mimicry of a nucleoprotein (NP) sequence from A/Puerto Rico/8/1934 (PR8) strain, the backbone used in the construction of the reassortant strain X-179A that was used in Pandemrix{\circledR} vaccine, and reported on anti-hypocretin (HCRT) receptor 2 (anti-HCRTR2) autoantibodies in narcolepsy, mostly in post Pandemrix{\circledR} narcolepsy cases (17 of 20 sera). In this study, we re-examined this hypothesis through mass spectrometry (MS) characterization of Pandemrix{\circledR}, and two other pandemic H1N1 (pH1N1)-2009 vaccines, Arepanrix{\circledR} and Focetria{\circledR}, and analyzed anti-HCRTR2 autoantibodies in narcolepsy patients and controls using three independent strategies. Methods: MS characterization of Pandemrix{\circledR} (2 batches), Arepanrix{\circledR} (4 batches) and Focetria{\circledR} (1 batch) was conducted with mapping of NP 116I or 116M spectrogram. Two sets of narcolepsy cases and controls were used: 40 post Pandemrix{\circledR} narcolepsy (PP-N) cases and 18 age-matched post Pandemrix{\circledR} controls (PP-C), and 48 recent (6 months) early onset narcolepsy (EO-N) cases and 70 age-matched other controls (O-C). Anti-HCRTR2 autoantibodies were detected using three strategies: (1) Human embryonic kidney (HEK) 293T cells with transient expression of HCRTR2 were stained with human sera and then analyzed by flow cytometer; (2) In vitro translation of [35S]-radiolabelled HCRTR2 was incubated with human sera and immune complexes of autoantibody and [35S]-radiolabelled HCRTR2 were quantified using a radioligand-binding assay; (3) Optical density (OD) at 450 nm (OD450) of human serum immunoglobulin G (IgG) binding to HCRTR2 stably expressed in Chinese hamster ovary (CHO)-K1 cell line was measured using an in-cell enzyme-linked immunosor-bent assay (ELISA). Results: NP 116M mutations were predominantly present in all batches of Pandemrix{\circledR}, Arepanrix{\circledR} and Focetria{\circledR}. The wild-type NP109-123 (ILYDKEEIRRIWRQA), a mimic to HCRTR234-45 (YDDEEFLRYLWR), was not found to bind to DQ0602. Three or four subjects were found positive for anti-HCRTR2 autoantibodies using two strategies or the third one, respectively. None of the post Pandemrix{\circledR} narcolepsy cases (0 of 40 sera) was found positive with all three strategies. Conclusion: Anti-HCRTR2 autoantibody is not a significant biological feature of narcolepsy or of post Pandemrix{\circledR} autoimmune responses.",
author = "Guo Luo and Ling Lin and Louis Jacob and M{\'e}lodie Bonvalet and Aditya Ambati and Giuseppe Plazzi and Fabio Pizza and Ryan Leib and Adams, {Christopher M.} and Markku Partinen and Mignot, {Emmanuel Jean Marie}",
note = "Ricercatori distaccati presso IRCCS a seguito Convenzione esclusiva con Universit{\`a} di Bologna (Plazzi Giuseppe, Pizza Fabio). Richiesto CORRIGENDUM per modifica di affiliazione (affiliazione imprecisa)",
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TY - JOUR

T1 - Absence of anti-hypocretin receptor 2 autoantibodies in post pandemrix narcolepsy cases

AU - Luo, Guo

AU - Lin, Ling

AU - Jacob, Louis

AU - Bonvalet, Mélodie

AU - Ambati, Aditya

AU - Plazzi, Giuseppe

AU - Pizza, Fabio

AU - Leib, Ryan

AU - Adams, Christopher M.

AU - Partinen, Markku

AU - Mignot, Emmanuel Jean Marie

N1 - Ricercatori distaccati presso IRCCS a seguito Convenzione esclusiva con Università di Bologna (Plazzi Giuseppe, Pizza Fabio). Richiesto CORRIGENDUM per modifica di affiliazione (affiliazione imprecisa)

PY - 2017/12/1

Y1 - 2017/12/1

N2 - Background: A recent publication suggested molecular mimicry of a nucleoprotein (NP) sequence from A/Puerto Rico/8/1934 (PR8) strain, the backbone used in the construction of the reassortant strain X-179A that was used in Pandemrix® vaccine, and reported on anti-hypocretin (HCRT) receptor 2 (anti-HCRTR2) autoantibodies in narcolepsy, mostly in post Pandemrix® narcolepsy cases (17 of 20 sera). In this study, we re-examined this hypothesis through mass spectrometry (MS) characterization of Pandemrix®, and two other pandemic H1N1 (pH1N1)-2009 vaccines, Arepanrix® and Focetria®, and analyzed anti-HCRTR2 autoantibodies in narcolepsy patients and controls using three independent strategies. Methods: MS characterization of Pandemrix® (2 batches), Arepanrix® (4 batches) and Focetria® (1 batch) was conducted with mapping of NP 116I or 116M spectrogram. Two sets of narcolepsy cases and controls were used: 40 post Pandemrix® narcolepsy (PP-N) cases and 18 age-matched post Pandemrix® controls (PP-C), and 48 recent (6 months) early onset narcolepsy (EO-N) cases and 70 age-matched other controls (O-C). Anti-HCRTR2 autoantibodies were detected using three strategies: (1) Human embryonic kidney (HEK) 293T cells with transient expression of HCRTR2 were stained with human sera and then analyzed by flow cytometer; (2) In vitro translation of [35S]-radiolabelled HCRTR2 was incubated with human sera and immune complexes of autoantibody and [35S]-radiolabelled HCRTR2 were quantified using a radioligand-binding assay; (3) Optical density (OD) at 450 nm (OD450) of human serum immunoglobulin G (IgG) binding to HCRTR2 stably expressed in Chinese hamster ovary (CHO)-K1 cell line was measured using an in-cell enzyme-linked immunosor-bent assay (ELISA). Results: NP 116M mutations were predominantly present in all batches of Pandemrix®, Arepanrix® and Focetria®. The wild-type NP109-123 (ILYDKEEIRRIWRQA), a mimic to HCRTR234-45 (YDDEEFLRYLWR), was not found to bind to DQ0602. Three or four subjects were found positive for anti-HCRTR2 autoantibodies using two strategies or the third one, respectively. None of the post Pandemrix® narcolepsy cases (0 of 40 sera) was found positive with all three strategies. Conclusion: Anti-HCRTR2 autoantibody is not a significant biological feature of narcolepsy or of post Pandemrix® autoimmune responses.

AB - Background: A recent publication suggested molecular mimicry of a nucleoprotein (NP) sequence from A/Puerto Rico/8/1934 (PR8) strain, the backbone used in the construction of the reassortant strain X-179A that was used in Pandemrix® vaccine, and reported on anti-hypocretin (HCRT) receptor 2 (anti-HCRTR2) autoantibodies in narcolepsy, mostly in post Pandemrix® narcolepsy cases (17 of 20 sera). In this study, we re-examined this hypothesis through mass spectrometry (MS) characterization of Pandemrix®, and two other pandemic H1N1 (pH1N1)-2009 vaccines, Arepanrix® and Focetria®, and analyzed anti-HCRTR2 autoantibodies in narcolepsy patients and controls using three independent strategies. Methods: MS characterization of Pandemrix® (2 batches), Arepanrix® (4 batches) and Focetria® (1 batch) was conducted with mapping of NP 116I or 116M spectrogram. Two sets of narcolepsy cases and controls were used: 40 post Pandemrix® narcolepsy (PP-N) cases and 18 age-matched post Pandemrix® controls (PP-C), and 48 recent (6 months) early onset narcolepsy (EO-N) cases and 70 age-matched other controls (O-C). Anti-HCRTR2 autoantibodies were detected using three strategies: (1) Human embryonic kidney (HEK) 293T cells with transient expression of HCRTR2 were stained with human sera and then analyzed by flow cytometer; (2) In vitro translation of [35S]-radiolabelled HCRTR2 was incubated with human sera and immune complexes of autoantibody and [35S]-radiolabelled HCRTR2 were quantified using a radioligand-binding assay; (3) Optical density (OD) at 450 nm (OD450) of human serum immunoglobulin G (IgG) binding to HCRTR2 stably expressed in Chinese hamster ovary (CHO)-K1 cell line was measured using an in-cell enzyme-linked immunosor-bent assay (ELISA). Results: NP 116M mutations were predominantly present in all batches of Pandemrix®, Arepanrix® and Focetria®. The wild-type NP109-123 (ILYDKEEIRRIWRQA), a mimic to HCRTR234-45 (YDDEEFLRYLWR), was not found to bind to DQ0602. Three or four subjects were found positive for anti-HCRTR2 autoantibodies using two strategies or the third one, respectively. None of the post Pandemrix® narcolepsy cases (0 of 40 sera) was found positive with all three strategies. Conclusion: Anti-HCRTR2 autoantibody is not a significant biological feature of narcolepsy or of post Pandemrix® autoimmune responses.

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