Balla-Jhagjhoorsingh, SS

Corti, D

Davis, D

Heyndrickx, L

Lanzavecchia, A

Vanham, G

Vereecken, K

Weiss, R

Willems, B

English

PubMed

SS Balla-Jhagjhoorsingh

B Willems

K Vereecken

L Heyndrickx

D Davis

D Corti

A Lanzavecchia

R Weiss

G Vanham

Springer - Publisher Connector

P04-52 LB. The importance of well-defined HIV-1 neutralization assays

BioMed Central
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Retrovirology
Open AccessPoster presentation
P04-52 LB. The importance of well-defined HIV-1 neutralization 
assays
SS Balla-Jhagjhoorsingh*1, B Willems1, K Vereecken1, L Heyndrickx1, 
D Davis2, D Corti3, A Lanzavecchia3, R Weiss4 and G Vanham1
Address: 1Institute of Tropical Medicine, Antwerp, Flanders, Belgium, 2Biomedical Primate Research Centre, Rijswijk, Netherlands, 3Institute for 
Research in Biomedicine, Bellinzona, Switzerland and 4University College London, London, UK
* Corresponding author    
Background
Plasma from HIV-1 seropositive individuals show a range
of specificities from variant specific to broad cross-neu-
tralizing. The plasma also show different kinetics of neu-
tralization which may be linked to their specificity. The
properties may be a consequence of being a polyclonal
mixture of antibodies. Isolating monoclonal antibodies
from these people would help to resolve these issues.
Methods
Panels of primary isolates of HIV-1 subtypes A, B, C and
circulating recombinant form CRF02_AG were prepared
in mitogen stimulated PBMCs. Neutralization assays were
performed with different lengths of incubation (1 h vs. 24
h), absorption (1 h vs. 24 h) and culture phases (7 days or
14 days for PBMCs and 2 days for cell lines). Plasma from
patients infected with either HIV-1 subtype C or
CRF02_AG and attending the HIV clinic in Antwerp were
screened in PBMC neutralization assays. Monoclonal
antibodies were then isolated from memory B cells of
individuals whose plasma cross-neutralized strains from
multiple subtypes in a high throughput HOS cell based
assay using pseudoviruses.
Results
The original donors' plasma were active against relatively
resistant HIV-1 isolates in assays with extended incuba-
tion phases. In contrast, the monoclonal antibodies were
active against relatively sensitive isolates in assays with
extended absorption phases. Pseudoviruses had a ten-
dency to be easier neutralized compared to their primary
viruses.
Conclusion
The neutralizing properties of the monoclonal antibodies
do not reflect those of the plasma from the original
donors. Either the method of selecting the monoclonals
or the proportion of monoclonals isolated may be a prob-
lem. Mixtures of monoclonals may be required for activity
against the relatively resistant isolates and the amount of
specific antibodies in the plasma is unknown. Alterna-
tively, there may be a discrepancy between specificities of
the bone marrow plasma cells and the circulating memory
B cells.
from AIDS Vaccine 2009
Paris, France. 19-22 October 2009
Published: 22 October 2009
Retrovirology 2009, 6(Suppl 3):P392 doi:10.1186/1742-4690-6-S3-P392
<supplement> <title> <p>AIDS Vaccine 2009</p> </title> <editor>Anna Laura Ross</editor> <note>Meeting abstracts - A single PDF containing all abstracts in this Supplement is available <a href="http://www.biomedcentral.com/content/files/pdf/1742-4690-6-S3-full.pdf">here</a>.</note> <url>http://www.biomedcentral.com/content/pdf/1471-2105-10-S12-info.pdf</url> </supplement>
This abstract is available from: http://www.retrovirology.com/content/6/S3/P392
© 2009 Balla-Jhagjhoorsingh et al; licensee BioMed Central Ltd. 


Lanzavecchia A

Corti D

Davis D

Heyndrickx L

Vereecken K

Willems B

Balla-Jhagjhoorsingh SS

Weiss R

Vanham G

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P04-52 LB. The importance of well-defined HIV-1 neutralization assays

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