Abstracts Vol. 1 [International Conference on AIDS (11th: 1996: Vancouver, Canada)]

Track A: Basic Science

Tu.A.2085 - Tu.A.2093

Methods: DNA samples were derived from HIV- I isolates in Korea. Some of those HIV- I
isolates were propagated in normal PBMCs, and DNA was extracted from expanded
PBMC cultures.The other HIV-I DNA samples were derived from PBMCs which were separated from HIV infected bloods. In all cases, viral envelope gene fragments(V3 region)
were amplified by polymerase chain reaction (I st PCR:V3 I/env 3 I, 2nd PCR: env 5 I/V3 3 I).
The 2nd PCR products, 340bp fragments were cloned and sequenced. By comparsion of
sequence data (DNAsis) and by phylogenetic analysis (neighbor joining method, MEGA),
subtypes of HIV- I isolates from Korean were identified.
Results: 1I0 HIV- I isolates of the studied I I HIV-I isolates belonged to subtype B. Between
them, 9 HIV- I isolates showed sequence diversity of 2-5%, and one isolates showed 10 -20%The other one was not characterized clearly because it showed low sequence homology(50-60%) with other isolates and reference strains.To get more clear informations about
HIV- I isolates, the analysis of gag gene fragments or the bigger env gene fragments are
required.
Conclusions: Although we studied small portion of HIV- I infected persons in Korea, we
suppose that subtype B is the representative HIV- I subtype in Korea.To get more exact
information about HIV subtype distributions in Korea, we need further studies on more
specimens with known epidemiological data.
Joo-Shil Lee. #5, Nockbun-dong, Unpyung-ku, Seoul, 122-020, Korea Telephone: 82-2-380 -I 696 Fax: [phone]
Tu.A.2085
GENETIC VARIABILITY OF HIV-I IN BENIN AND EVIDENCE OFA HIV-I
GROUP O - GROUP M DUAL INFECTION.
Janssens W, Heyndrickx L, Alary M*, Nkengasong J, Fransen K, Willems B, Baganizi E*, Joly J*,
Guedemd A**, Davo N***, van der Groen G. Institute ofTropical Medicine, Antwerpen,
Belgium:; *Centre de recherche, Hfpital du St-Sacrement, Canada; **CREDESA, Pahou,
Benin; **Programme National de Lutte contre le Sida, Cotonou, Benin.
Objective:To study the genetic diversity of HIV- I strains circulating in Cotonou, Benin in
1993-1994.
Materials and Methods: From 2 I HIV- I seropositive Ghanese and Togolese prostitutes living
in Cotonou, Benin, the env C2V3 region (300bp) was amplified from serum by RT-PCR,
directly sequenced and phylogenetically analysed. 142 HIV-I seropositive individuals, being
either prostitutes (n= 128), pregnant women (n=8) or persons visiting STD hospitals (n=6),
were analysed for HIV- I group O antibodies by a synthetic V3 loop peptides based ELISA.
Reactive samples were retested in a line immunoassay (LIA, Innogenetics, Belgium, for
research purposes only) and specific "in house" group O (ANT70, CA9,VI686) western
blots. Specific PCR primers in the pol gene were used to discriminate between group O0
and group M infections (Janssens W, et al. 1995.The Lancet, 346, 451I-452).
Results: Out of 2 I HIV- I sera, 19 were phylogenetically classified in group M as subtype A,
and two were classified as subtype G. Out of 142 seropositive individuals, one pregnant
woman had serological indications for a mixed group O/M infection: reaction in group O V3
loop ELISA, reaction with group 0 ANT70V3 loop peptide and group M V3 consensus and
gp4 I peptides in LIA, and antibodies to p66, gp4 I, p33, p24 on group 0 western blot.This
was confirmed by a positive PCR with as well group 0 and group M specific pol primers.
Additional evidence was obtained by sequencing and phylogenetic analysis of the amplified
pol fiagments (267bp).
Conclusion:This is the first report indicating that double infection with HIV- I group O and
M infection may occur Benin is besides Cameroon, Gabon, Equatorial Guinea and Nigeria,
the fifth African country where group O infection is recognized.
Wouter Janssens, Institute of Tropical Medicine, Nationalestraat 155, 2000 Antwerpen,
Belgium; Tel. [phone]; Fax: [phone]
Tu.A.2086
SPECIFIC DIAGNOSIS OF HIV- I GROUP O BY RESTRICTION ANALYSIS OF THE
POL GENE FRAGMENT.
Heyndrickx L, Janssens W, Fransen K, Nkengasong J,Vereecken K, Loussert-Ajaka I*, Girtler
L *. van der Groen G. Institute ofTropical Medicine, Antwerpen, Belgium; *Hfpital BichatClaude Bernard, Paris, France; **University of Munich, Munchen, Germany
Objective: To develop a specific diagnostic tool to monitor an HIV- I group 0 infection.
Material and Methods: Over all, 79 isolates including 23 HIV- I group 0, 2 SlVcpz, and 54
HIV- I group M (subtypes A-H) viruses were analysed, by using highly sensitive (93%) and
specific (100%) diagnostic primers in the I-l gene for nested PCR of HIV- I infected primary lymphocytes 1,2. We obtained fragments of respectively 175 and 239 bp long. After treatment with a ricstriction enzyme (Pst I) the PCR fragment is electrophoresed on apolyacrylaimide gel (7.5%) and stained with ethidiumbromide.
Results: For the pl fragments belonging to HIV- I group 0 and the two SIVCPZ isolates, a
unique Pt I site e was present with exception of one fragment belonging to an HIV-I group
O isolate (BCF08), generating fragments of 107 bp + 68 bp or 107 bp + 132 bp (depending on the fiagment used). For the pol fragments belonging to HIV- group M no such
restriction site was present resulting in only one band of 175 bp. Restriction analysis of psi
(RAP) is much faster and cheaper as phylogenetic analysis of sequences.
Conclusion: Restriction analysis of a I175 bp or 239 bp long fragment of J is a promising
fast and simple (no sequencing) method to confirm an HIV- I group 0 infection.
Leo Heyndrickx, Institute of Tropical Medicine, Natiunalestraat I155, 2000 Antwerpen,
BelgiuinTol: 32 3 247 63 26, Fax: [phone]
Tu.A.2087
HUMAN IMMUNODEFICIENCY VIRUS VARIANTS IN CUBA.
Rolo Felipe1, Blanco M I, Lobaina LI, Cassol S2, Balotta C3, Gomez C4, Duarte C4. Martin
Z t, Gaili M3, Machado F, 0) Shaugnessy M2. I -National AIDS Res. Lab. Cuba. 2-B. C.
Cent-e for Excellence in HIV/AIDS.Vancouver. Canada. 3-Lab. lnf. Dis. Clinic. Milan. Italy. 4 -C.I.G.B. Cuba.
Objective: To identify the HIV- I C2V3 region sequences in Cuban patients.
Methods: The peripheral blood mononuclear cells (PBMC) from six young people who
weire infected in Cuba, were lysed, the DNA was obtained and further amplified in two diffceet laboratories by nested polymerase chain reaction (PCR) using different primers pairs

for the C2V3 region.The PCR products were cloned and sequenced in an automatic
sequencer from Applied Biosystem.
Results: The results of both laboratories were in agreement and they showed a distinct
tetramer that is GRGR in the tip of the V3 loop.There was evidence of transmission in two
of these young people who were sexually related. As we could appreciate by the phylogenetic analysis and the epidemiological information, they were all probably related.
Conclusions: We identified an uncommon amino acid in the tip ofV3 loop, the GRGR
motif has not been previously reported.We discuss the possibility of a phenomenon of
convergence or simply the presence of a different lineage for subtype B in the HIV strains
population in Cuba.
Felipe Rolo, Lab. de Investigaciones del SIDA. Ministerio de Salud Publica 23 y N Vedado
Habana. Cuba. Fax [phone]Teldfono: 537 064 62162.
Tu.A.2088
HIV- I VIRAL CHARACTERISTICS, SERUM NEUTRALIZING ACTIVITY AND DISEASE
PROGRESSION IN A GROUP OF HIV+ PATIENTS
Massetti A. Paola, *Ercoli L, Galati V., *Nicastri E., d'Ettorre G., *Ciapetti C., Marchese R.,
VulloV., *Andreoni M. Dept. Inf.Trop. Dis., La Sapienza Univ. and *Chair Inf. Dis.,TorVergata
Univ., Rome, Italy
Objective: To evaluate virological and immunological parameters which might influence HIV
progression.
Methods: The study was performed on 35 patients with CD4 cell count <250/mmc, showing a different progression rate during a follow up of >2 years: 18 patients remained clinically and immunologically stable and 17 showed signs of progression.The 2 groups were
homogeneous for sex, risk factor, age, follow up period and mean CD4 cell count at start of
the study; all patients were treated with zidovudine.The following parameters were studied:
replication rate of plasma HIV strains, plasma viremia titer, viral load measurement by HIVRNA quantitative PCR, MT-2 syncytium assay determination of codon 21 5 reverse transcriptase mutation, zidovudine sensitivity neutralizing activity
Results:The results obtained in the 2 groups of patients are shown in the table (*p<0.005;
**p<0.00 I).
Non-progressors (18) Progressors (17)
Plasma isolation rate 88.8% 100%
Median plasma isolation time (days)* 22. I 15.2
Mean plasma viremia titer (TCID50/ml)* 15 1 030
Mean HIV-RNA copy number/ml** 129000 465000
Syncytium inducing phenotype 68.7% 88.2%
Codon 215 mutation 68.7% 88.2%
Zidovudine resistant strains 61.5% 73.3%
Neutralizing activity in serum* 53.8% 26.7%
Conclusions: AZT-treated patients with CD4 <250/mmc and without signs of clinical
and/or immunological deterioration have plasma viral titer and HIV-RNA copy number significantly lower than progressed patients. Our results seem to indicate that this could be
mainly due to a different response to the infection (higher neutralizing activity in serum of
non-progressors) than to the biological characteristics of the infecting strain.
A. Paola Massetti, Dept. Infectious and Tropical Diseases, La Sapienza University Policlinico
Umberto I,Viale del Policlinico 155, 00161 Rome, ItalyTel. 39 6 491749 Fax: [phone]
Tu.A.2092
STRUCTURAL PECULARITIES OF HIV AT THE EARLY STAGE OF HIV/AIDS EPIDEMIC
IN ST.PETERSBURG, RUSSIA.
A.EmeljanovI, A.Malykh2, O.Barabitskaya2, H.Farzadegan3, M.C.Reitz,Jr2, A.Kozlov1. I
Biomedical Center for AIDS, Cancer and Related Problems, Research Institute of Pure
Biochemicals, St.Petersburg, Russia. 2 Laboratory of Tumor Cell Biology, National Cancer
Institute, National Institute of Health, Bethesda, Maryland 20892 3 Johns Hopkins University,
School of Public Health, Baltimore, Maryland 21218
Objectives: To study whether structural peculiarities of HIV exist at the early stage of
HIV/AIDS epidemic in Russia
Methods: An approach of nested PCR with multiple pairs of primers was used to amplify
and sequence env-specific fragments from HIV-infected patients at the early stages of infection (CDC: liIA-liB).
Results: Analysis of nucleotide and amino acid sequences has shown both similarities with
already published sequences and some peculiarities.The latter include high mutation fiequency in C2 and C3 regions and high variability in amino acids' positions 18, 25 and 29 of
V3. Sequences APGS, APGG and HMGPGGTF at the apex ofV3 in the St. Petersburg isolates ru38 18, ru426I and ru154 were not found in HIV Sequences Database. Molecular
biological data suggest that St. Petersburg isolates belong to low/slow, NSI/monocytotropic
phenotype.The intrasubtype genetic diversity is characterised by a broad distribution with
median II,67%.
Conclusions: Structural peculiarities of the St Petersburg isolates as well as the broad intrasubtype genetic diversiny (with median I,67%) may constitute peculiarity of the early stage
of HIV/AIDS epidemic in Russia.
The study was supported by Grants NoR I B000 and NoR I 8300 from International Science
Foundation and Grant NoSD3TV00010 from the togarty International Center.
A.Emeijanos, Lab of Molecular Virology, Institute of Pure Biochemicals, 7, Pudozskaja St.,
St.Petersburg, Russia I 97 I I O.Te.: 7-(81I2)-21t0-7869, Fax: 7-(81I2)-230-4948, E-mail:
hivaidsimod.hpb.spb.ru
Tu.A.2093
HIV-l GROUP 0 INFECTION IN SPAIN
Soriano Vincent, Garcia-Lerma G, Mas A, Gutidrrez M, Bravo R, Heredia A*, Pdrez-Labad
MaL, Hewlett I*, Domingo E**,GOonz~lez-Lahoz J. CIC. Instituto do Salad Carlos Ill, Madrid.
**Lab. Mlecular Virology, CBER, FDA, Bethesda, MA, USA. **CBM, Universidad Autmail:
[email protected].
Introduction: HIV- I group 0 was first isolated in I1987. It has distinct serological behaviour

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