Bridging the Gap: Conference Record [Abstract book, International Conference on AIDS (12th: 1998: Geneva, Switzerland)]

12th World AIDS Conference

Abstracts 60314-60318 1057

LP-BM5 murine leukemia virus, were used as MAIDS mice. One day after the
infection of C. neoformans (strain 613D, 1 x 106 cells/mouse), MAIDS mice were
treated with 10 mg/kg dose of Z-100 once every other day for 10 injections. A
mixture of type 2 cytokines (IL-4 and IL-10) was injected at 50 ng/mouse to normal
mice infected with C. neoformans. The decrease in numbers of C. neoformans
cells in brain tissues of MAIDS mice treated with Z-100 was considered as a result
of the antifungal activity of the compound. The amount of fungus in brains were
measured by the plating of 10% homogenized brain tissues on Sabouraud agar
plates. The amounts of cytokines (IL-2, IL-4, IL-10 and IFN-y) in culture fluids of
spleen cells (5 x 106 cells/ml) from MAIDS mice treated with or without Z-100
were measured by ELISA.
Results: The growth of C. neoformans was markedly suppressed in brains of
MAIDS mice treated with Z-100, as compared with those of MAIDS mice treated
with saline. Conversely, the growth of C. neoformans was increased in brains
treated with a mixture of type 2 cytokines. After stimulation with anti-CD3 mAb
(10 pg/ml), splenic lymphocytes from MAIDS mice produced type 2 cytokines into
their culture fluids. However, these cytokines were not demonstrated in culture
supernatants of splenic cells from MAIDS mice treated with Z-100.
Conclusion: Dissemination of C. neoformans in MAIDS mice should be influenced by type 2 T cell response stimulated by LP-BM5 virus infection, and
through the regulation of these type 2 T cell responses, Z-100 may inhibit the
growth of C. neoformans in the brains of these mice.
60314 Pathogenic roles of type 2 T cell responses on
Cryptococcus neoformans infection in MAIDS mice
Katsunori Furukawa, H. Sasaki, R.B. Pollard, F. Suzuki. University of Texas
Medical Branch, Galveston, Texas 77555-0835, USA
Objective: Cryptococcus neoformans (C. neoformans) has been reported as a
severe pathogen in AIDS patients, though it rarely disseminates in healthy individuals. Since T cells play an important contribution on the host's defense against
this infection, a pathogenic role of type 2 T cell response (T2 response) on the
severity of C. neoformans infection was investigated in this study.
Methods: C57BL/6 mice (4-weeks-old) were exposed to LP-BM5 murine
leukemia virus (4.5 x 102 PFU/mouse). Fifty days after infection, these mice
(MAIDS mice) were infected intratracheally with C. neoformans, strain 613D
grown for 2 days in Sabouraud dextrose agar. The 10% homogenates from lungs
and brains removed from these mice 3 weeks after infection were prepared, and
subjected to the colony forming assay. T2 responses were determined as the titer
of IL-4 and IL-10 in culture fluids of splenic cells derived from infected MAIDS
mice. The cytokines produced by splenic cells after stimulation with anti-CD3
mAb were determined by ELISA.
Results: As compared to normal mice, the severity in the growth of C. neoformans was increased greatly in MAIDS mice. A predominance of T2 response was
demonstrated in MAIDS mice. The growth of C. neoformans in MAIDS mice was
reduced to the levels observed in normal mice, when they were treated with a
mixture of mAbs against type 2 cytokines. Conversely, the susceptibility of normal
mice to infection with C. neoformans was increased when they received a mixture
of type 2 cytokines.
Conclusions: In this study, an important role of type 2 T cell responses on the
severity of C. neoformans infection is demonstrated. In MAIDS mice, the growth
of C. neoformans was accelerated by type 2 T cells induced by the association
with LP-BM5 murine virus infection.

Conclusion: Representative seroepidemiologic studies performed in "more
exposed populations" are very helpful to design adequate preventive health policies. In this regard, STD clinics, mutually connected and wih the public hospital
network, are highly appropriate centers for identifying subjects.
S60316 Reverse transcriptase and protease genotypes in
patients with HIV-1 symptomatic primary infection on
antiretroviral tritherapy (ANRS053 trial)
Martine Harzic1, M. Denayrolles2, B. Dumon2, B. Dubeaux', I. Pellegrin2,
E. Mornet3, H. Fleury2. 1 C.H. Versailles, Hopital A. Mignot, Sce De
Microbiologie, 78157 Le Chesnay; 2Hopital Pellegrin, 33-Bordeaux; 3Sesep
Universite de Versailles, 78-Versailles, France
Objective: To study reverse transcriptase (RT) resistance mutations (codons 184
and 215) and viral protease polymorphism in patients with symptomatic primary
HIV-1 infection on highly active antiretroviral therapy (HAART): zidovudine (ZDV),
lamivudine (3TC) and ritonavir.
Design: Analysis was done on the plasma at day 0 (DO) for RT and protease
genes in 41 patients among 61 enrolled in the ANRS053/053B trial. On follow up
(6 to 18 months) analysis of RT gene was performed in 23 patients from plasma
or peripheral blood lymphocytes.
Methods: RT gene was analysed by a differential hybridization assay using
probes specific for the codons 184 and 215. Protease viral gene was analysed
by sequencing.
Results: At DO, 2 patients out of 41 were infected by RT mutated viruses:
one with T215Y/F mutation and another with M184V mutation. Protease gene
polymorphism was mainly located in 3 regions (codons 10 to 19, codons 33 to
41 and codons 57 to 77). Key mutations associated with resistance to ritonavir
were detected in three samples: one with a mutation V361, one with a mutation
A71V and one with mutations V361 and A71T. On follow up, analysis on the RT
gene did not show any new resistance mutations to AZT and/or 3TC, even in 4
patients with persistent HIV-1 replication (plasma HIV-1-RNA > 1000 copies per
mL) who were poorly compliant to therapy.
Conclusion: Prevalence of genotypic resistance to ZDV, 3TC and ritonavir is
low in patients with primary HIV-1 infection.
Persistence of a viral replication in some patients was not explained by the
development of known resistance mutations to used drugs.
60317 Genotypic analysis of the HIV-1 reverse transcriptase
gene using differential hybridization
Martine Harzic, P. Sagert, B. Dubeaux, M. Joannes, F. Doucet Populaire,
J.C. Ghnassia. C.H. Versailles, Hopital A. Mignot, SCE De Microbiologie, 78157
Le Chesnay, France
Objective: To develop a differential hybridization assay on microplates to detect
mutations in the codons 184 and 215 of the HIV-1 reverse transcriptase gene.
Methods: Culture supernatants of wild or mutated HIV-1 strains were used to
develop the method. Viral RNA was amplified by nested RT-PCR. Amplified products were coated on microplates using Hybridowell' (Argene-Biosoft, France)
and hybridized with biotinilated probes. Different probes, specific for the wild or
mutated, 184 or 215, codons were tested. Forty plasma samples from HIV-1
infected patients were analysed by this method. Results were compared with
those obtained by selective PCR assay using two different antisens primers, each
with a 3'nucleotide specific for the wild or the mutated codon.
Results: Culture supernatants: All the amplified products were detected by
hybridization with one of the two probes with a 100% specificity. Analysis of
amplified product obtained from a viral population of two different strains allowed
detection of a minor population of 12.5%.
Plasma: In 30 samples there was a single viral population detected by both
methods. In ten samples there was a double, wild and mutated, viral population.
In that case discordant results were obtained for 4 samples. This could be due to
a known lack of selective PCR specificity.
Conclusion: This differential hybridization assay is rapid and reproducible. It
allows a good discrimination between wild and mutated populations and detect a
12.5% minor viral population.
|60318 HHV-8 sequence variability in French HIV-1 infected
patients
Bernard Masquelier1, Franck Boralevi', P. Caminade1, M. Denayrolles1,
C. Cazorla2, M. Dupon2, H.J.A. Fleury'. 'Laboratoire de Virologie Chu Bordeaux
Place Amelie Raba Leon 33076 Bordeaux Cedex; 2Departement D'lnfectiologie
Chu Bordeaux, Bordeaux, France
Objectives: To study the genotypic variability of HHV-8 in France and to correlate
it with epidemiological and clinical data.
Methods: 33 HIV-1 infected patients followed-up in Bordeaux University
hospital, France, had AIDS-Kaposi's sarcoma and underwent skin biopsies. One

1603151

Evolution of HIV seroprevalence in intravenous drug
users, homosexual/bisexual males and prostitutes in
Madrid area (1986-1998)

Jorge Del Romero Guerrero, S. Garcia, J. Ballesteros, I. Clavo, D. Carrio,
B. Baza, C. Rodriquez. Centro Sanitario Sandoval (Comunidad De Madrid),
C/Sandoval, Num. 728010-Madrid, Spain
Objective: To know the evolution over time of the prevalence of HIV infection in
more vulnerable groups in the Autonomous Community of Madrid.
Study Population: From March 1986 to January 1998, 22.775 subjects
(14.567 males and 8.208 females) first attended an STD/HIV clinic of the Autonomous Community of Madrid in order to know their serological HIV status.
6.419 of them were homosexual or bisexual males (HM/BI), 3.924 were intravenous drug users (IDU), and 1.777 women non-IDU prostitutes.
Methods: All these patients completed an anonymous epidemiological questionnaire ir order to identify risk practices for HIV and use the adequate information
to change risk behaviour for this infection. HIV serology testing was also performed
(ELISA+WB).
Results: The evolution of HIV annual seroprevalence in the study groups is
shown in the table below. From 1988, a significant steady decrease in requests
for HIV tests occurs in IDU. Women non-IDU prostitutes showed infection rates
of about 1% in the last five years. A steady increasing mean age was observed
among HIV-positive patients, as much in IDU (25 years in 1986 and 32 in 1997)
as in HM/BI (30 years in 1996 and 35 in 1997).

60315: Table
1986 1987
IDU n = 3.924 58.1 56.4
HM/BI n= 6.419 19.7 18.3
Prost. n =1.777 0 2.9

1988
51
24.6
1.7

1989
58.9
25.1
3.4

1990
51.1
29.7
5

1991
54.5
25.7
1.5

1992
52.4
21.9
4.1

1993
45.9
17.1
1.1

1994
45.5
12
1.3

1995
50.4
15.3
1.1

1996 1997
41.8 44.4
12.5 11.7
1.1 0.5

Total
2058/3924 (52.4%)
1206/6419 (18.8%)
27/1777 (1.5%)