INVESTIGADORES
FERNANDEZ LARROSA Pablo Nicolas
congresos y reuniones científicas
Título:
ENHANCEMENT OF HIV-1 VIRAL PRODUCTION IN VITRO BY SUPERINFECTION.
Autor/es:
P. N. FERNÁNDEZ LARROSA; A. CEBALLOS; R.D. RABINOVICH; S. MARQUINA; L. A. MARTÍNEZ PERALTA
Lugar:
Paris, France
Reunión:
Conferencia; 2st IAS Conference on HIV pathogenesis and treatment; 2003
Resumen:
Superinfection with HIV-1 occurs in vitro and in vivo, and is
considered to be the main cause of the high frequency of circulating
recombinant forms worldwide. Viral production is normally high in
patients and different viral variants accumulate over time causing the
necessary conditions for superinfection. Previous experiments in our
laboratory suggested pseudotype production due to superinfection with
different HIV-1 strains. However, no dynamic studies about viral
production have been made in a superinfection system. In order to study
the influence of superinfection on viral production, H9-HTLVIIIB cells,
infected with HXB2 viral strain, were pretreated with 2.2 microg/ml
polybrene for 30 min at 37$#176;C and centrifugated at 4$#176;C, before
being superinfed with MN strain at moi=1. Viral production was
quantified by syncytia production using MT2 cells by limiting dilution,
and by a commercial ELISA for p24 antigen (HIVAGTM-1 Monoclonal Abbott,
Abbott Laboratories). The virus produced by the superinfected cells was
used to infect MT2 cells and characterization of proviral DNA in MT2
cells was made by amplification of the C2V3 region of env gene followed
by digestion with an enzime (BsmA I) that differentiates both viral
strains. Syncytia production revealed a 1.7 log increase at the first
day post-superinfection (ps) as regard to a non superinfected
H9HTLVIIIB control, while p24 antigen quantification suggested a 1.5
increase. Viral production during following days did not differ
significantly from the control. Viral characterization showed both
strains production at first day, while at following days, only the
original strain (HXB2) was detected. Superinfection in vitro of
H9HTLVIIIB cell line with HIVMN strain increased total viral production
only at first day post-superinfection. The cytophatic effect observed
since 24 h ps could probably explain why this enhancement was not
observed later on. This viral mechanism could play an important role in
vivo in the phisiophatology of HIV infection and persistence.