tetano
Editor, Senior Moderator
PLoS One. 2015 Aug 12;10(8):e0135383. doi: 10.1371/journal.pone.0135383. eCollection 2015.
[h=1]An Innovative Pseudotypes-Based Enzyme-Linked Lectin Assay for the Measurement of Functional Anti-Neuraminidase Antibodies.[/h] Prevato M[SUP]1[/SUP], Cozzi R[SUP]2[/SUP], Pezzicoli A[SUP]2[/SUP], Taddei AR[SUP]3[/SUP], Ferlenghi I[SUP]2[/SUP], Nandi A[SUP]4[/SUP], Montomoli E[SUP]5[/SUP], Settembre EC[SUP]6[/SUP], Bertholet S[SUP]2[/SUP], Bonci A[SUP]2[/SUP], Legay F[SUP]7[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Antibodies (Ab) to neuraminidase (NA) play a role in limiting influenza infection and might help reduce the disease impact. The most widely used serological assay to measure functional anti-NA immune responses is the Enzyme-Linked Lectin Assay (ELLA) which relies on hemagglutinin (HA) mismatched virus reassortants, or detergent treated viruses as the NA source to overcome interference associated with steric hindrance of anti-HA Ab present in sera. The difficulty in producing and handling these reagents, which are not easily adapted for screening large numbers of samples, limits the routine analysis of functional anti-NA Ab in clinical trials. In this study, we produced influenza lentiviral pseudoparticles (PPs) containing only the NA antigen (NA-PPs) with a simple two-plasmid co-transfection system. NA-PPs were characterized and tested as an innovative source of NA in the NA inhibition (NI) assay. Both swine A/California/07/2009 (H1N1) and avian A/turkey/Turkey/01/2005 (H5N1) N1s within NA-PPs retained their sialidase activity and were specifically inhibited by homologous and N1 subtype-specific, heterologous sheep sera. Moreover, A/California/07/2009 N1-PPs were a better source of NA compared to whole live and detergent treated H1N1 viruses in ELLA, likely due to lack of interference by anti-HA Ab, and absence of possible structural modifications caused by treatment with detergent. This innovative assay is safer and applicable to all NAs. Taken together, these results highlight the potential of NA-PPs-based NI assays to be developed as sensitive, flexible, easy to handle and scalable serological tests for routine NA immune response analysis.
PMID: 26267900 [PubMed - in process] Free full text
[h=1]An Innovative Pseudotypes-Based Enzyme-Linked Lectin Assay for the Measurement of Functional Anti-Neuraminidase Antibodies.[/h] Prevato M[SUP]1[/SUP], Cozzi R[SUP]2[/SUP], Pezzicoli A[SUP]2[/SUP], Taddei AR[SUP]3[/SUP], Ferlenghi I[SUP]2[/SUP], Nandi A[SUP]4[/SUP], Montomoli E[SUP]5[/SUP], Settembre EC[SUP]6[/SUP], Bertholet S[SUP]2[/SUP], Bonci A[SUP]2[/SUP], Legay F[SUP]7[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Antibodies (Ab) to neuraminidase (NA) play a role in limiting influenza infection and might help reduce the disease impact. The most widely used serological assay to measure functional anti-NA immune responses is the Enzyme-Linked Lectin Assay (ELLA) which relies on hemagglutinin (HA) mismatched virus reassortants, or detergent treated viruses as the NA source to overcome interference associated with steric hindrance of anti-HA Ab present in sera. The difficulty in producing and handling these reagents, which are not easily adapted for screening large numbers of samples, limits the routine analysis of functional anti-NA Ab in clinical trials. In this study, we produced influenza lentiviral pseudoparticles (PPs) containing only the NA antigen (NA-PPs) with a simple two-plasmid co-transfection system. NA-PPs were characterized and tested as an innovative source of NA in the NA inhibition (NI) assay. Both swine A/California/07/2009 (H1N1) and avian A/turkey/Turkey/01/2005 (H5N1) N1s within NA-PPs retained their sialidase activity and were specifically inhibited by homologous and N1 subtype-specific, heterologous sheep sera. Moreover, A/California/07/2009 N1-PPs were a better source of NA compared to whole live and detergent treated H1N1 viruses in ELLA, likely due to lack of interference by anti-HA Ab, and absence of possible structural modifications caused by treatment with detergent. This innovative assay is safer and applicable to all NAs. Taken together, these results highlight the potential of NA-PPs-based NI assays to be developed as sensitive, flexible, easy to handle and scalable serological tests for routine NA immune response analysis.
PMID: 26267900 [PubMed - in process] Free full text