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

Tu.A.2067 - Tu.A.2072

Tuesday July 9, 1996

Results: Our sequence data confirm arid extend the previous notion that,,,ytur o nduc
ing (SI) variants with basic amino acid substitutions at particular positions ir V c3 evolve and
become a distinct population during disease progression. Elongation ofV2 vas furthier
noticed in 12 cases, which was most likely caused by duplication of a sequenc e of 9, 12:r
15 nucleotides.There appeared to be a strong correlation between V2 elon itirn nd rna I
erate/slow disease progression, since all the analyzed patients with a modirate/slow rate of
disease progression featured elongated V2, while rapid progressors so far did not.The nai ii
feature for the latter was basic amino acid substitutions in V3.
Conclusions: Our results suggest the association of particular sequcie leaturer f both V3
and V2 with particular features of disease progression. Elongation of V2 loop nay I>e a good
predictor of moderate/slow disease progression, while basic substitutions of V3 ithout
elongation of V2 is characteristic of rapid progression.
T. Shioda,4 4-6 I Shiroganedai, Minato ku,Tokyo 108, Japan telephone: 8 I 3-5449-5282
Fax:[phone]
Tu.A.2067
IDENTIFICATION OF THE PRESENCE OF SELECTIVE PRESSURES ACTING ON
ENVELOPE C2/V3 REGION IN HIV- I SUBTYPE E INFECTED ASYMPTOMATIC
CARRIERS
Sato, Hironori*,Taniguchi K*.Tomita Y*, Miyakuni-*, Foongladda 5*'*, OVasi C' '+,rlskebe
Y*. *National Institute of HealthTokyo, Japan; * Naha Prefectural Hospital, Olkinawa, Japan;
** Mahidol University Bangkok,Thailand
Objective: Genetic characterization of HIIV- I subtype E strains undergoing person to person transmission and persistent infection.
Methods: We determined envelope C2/\V3 nucleotide sequences (324bp) >f I IIV I subtype
E proviral DNAs derived from an epiderniologically-linked intrafamilial infection case in
Japan and from 2 pairs of asymptomatic couples in Thailand. Blood specirmens vwere collected between June 1993 and September 1995. Extent of sequence variation in envelope C2
(125 bp),V3 loop (105 bp) and post-V3 loop regions (94 bp) was compar 'd in an each
individual or among individuals.
Results: In a virus-transmitter of the Japanese family V3 loop sequences were more heterogeneous than other regions in the C2/V3 region (P < 0.0001), while in two other asyrnpto
matic virus-recipients, the V3 loop sequences were less heterogenreous than post V3 region
(P < 0.000 I). Among other 2 asyrnptomatic couples in Thailand, one for each couple,
showed less heterogeneity in the V3 loop than in the post-V3 region (P < 0.0001). In all
cases, C2 region was highly homogeneous.The V3 loop sequences which showed less het
erogeneity than post V3 in an individual were sirnilar among different individuals as well as
to sequences fiorn asymptomatic carriers sampled between 1990 and 1992 in thailand,
while post-V3 loop sequences were highly divergent among them. These V'3 loops all
retained a potential glycosylation site and low net-positive charge in a V3 loop.
Conclusions: Unexceptional homogeneity of the C2 region suggest a critical roe of thtis
region in functional envelope formation. In contrast,V3 loop honmogeneity limited in one of
asymptomatic pair suggests a selective pressure which reduces sequence heterogeneity in
either viral transmission or asymptornatic persistence. Sequence similarity among the selected V3 loop sequences mnay suggest a selective advantage of the particular subset ofV3 loop
sequences.
H. Sato, AIDS Research Center, National Institute of HIealth,-loyarn,, I 2 1 I, Shiritiku.u,
Tokyo 162, Japan.Tel: [phone] III ext. 2532; Fax: (81)- 135285-1177; Email:
[email protected]
Tu.A.2068
HIGHLY DIVERGENT env SEQUENCE OF HIV- I B SUBTYPE AND DUAL INFECTION
OF HIV-I B AND E SUBTYPES
Xin K- *, Cao X-Ri, Shapshak P"*, Crandall KA*i*, Nislhoka K * *, Bukawu ItI, IHareajinr
K*, Fukushima J*, KanekoT*, Kawamoto S*, Okudaci K*. 'Yokohama City lnnv.Yokohara,
Japan; "Univ. MiMiami,iami, FL.;**Univ.Texas, Austin,TX; **Viral Hepatitis Research
Foundation of Japan.
Objects: Extensive heterogeneity of HI-lV is a major issue of I 1/ vaccine i'rrlopni ntTic he
ideal HIV vaccine should cover all HI-V subtypes and most of HIV isolates. In this,ibstract,
we report highly divergent env sequences of H IV I B subtype derived forno patient A frore
Miami, Florida, and dual HIV- I B and E subtypes infect patient B fron 1 Thai.
Materials and methods: Genomic DNA was extracted from peripheral blood monironucleari
cells (PBMC) of patient A and from the PBMC co culture of patient B. HIV I envelope gene
was amplified by polymerase chain reaction (PCR). Nested amplified PCR product', were
directly subcloned into pCRII vector Ten and thirteen clones were sequenced from patient
A and B, respectivelyTwo phylogenetic trees were constructed from necleotide sequences
of the env C2 V3 domain using the neighbor joining algorithm.
Results: The sequences derived fromi patient A are menmbers of the HIVI B suutype: how
ever, they atre highly divergent from known sequences of the HIV- I B subltype.The
sequences derived fr-om patient B demonstrated two distinct HIV subtypes, B and E.
Discussion and conclusion: to our knowledge, this is the first repcort of dual HIV I suiltype'
infection.The character of highly divergent env sequences is needed flture study.
K-Q Xrn, Dept. BactenoIYokoharn City Un. 3 9 1-ukuura, Kanazawa-ku,Yuiw ohtmY',i 236,
Japan. Eel. 045-787-2602 Fax: [phone],
Tu.A.2069
COMPARISON AMONG THE GENETIC DIVERSITIES OF WHOLE AND INFECTIOUS
HIV-I POPULATIONS IN BOTH PLASMA AND PERIPHERAL BLOOD MONONUCLEAR CELLS
Kato. Shingo', Hiraishi Y", Sugita T, Asakawa M*, Iranabusa Hl',tilsarkto T*. Keio
University Tokyo, Japan; "Ogikubo Hospital, lokyo, Japan
Objective:To investigate the genetic differences between HIV I popurations in plasma and
in peripheral mononuclear cells (PBMC) and between infectiorss and whole HIV I populations.
Methods: Plasma and PBMC were obtained from three HIV I positive patients A, B, and C.
HIV I RNA in plasma and HIV I DNA in PB1-C were clonally amplifed by PCR. Infectious
HIV- I in plasma and PBMC were cloned by the plaque hybridization assayThe enyV3
regions of ten clones from each H-IV- I population were sequenced Unrooted phylogenetic

trees were created from the sequence data of all HIV I clones obtained from each patient.

Since the unuer 4in..fe'.tious IHIV I clones obtained from plasma of patients A and C
were far less tfhan 10, the genetic analysis of these HIV- I populations was not carried out.
Results: In ptient A,iti no symptom and CD4 count of 544, the clones from the whole
HIV I populaticnr i iasian were more diverged than those in PBMC, and the clones froom
the infectious Hi i popution in PBMC were the most homogeneous. In patienrt B with
AIDS and CD4 cou it of. the clones from the four HIV-1 populations (whole and infectious HIV- I in both pla, r-,,i and PBMC) were equally homogeneous. In patient C, fiom
whom HIV I clones were isolated five times consecutively over 2.2 years (CD4 count was
changed from 272 to 131), the clones were clustered into six groups on the phylogenetic
analysis. One of the groups was specific for the HIV- I population in plasma, and another
was specific for the whole HIV I population in PBMC.T-he clones from the infectious HIV I
population in PBMC almost fell into one group.
Conclusions: Iie genetic characteristic of HIV-I qucasispecies in vsivo i quite different in
plasma and PBM( and depends on infectivity.-he genetic diversity of infectious HIV- I pop
ulation is not higher than that of whole HIV I population and tends to decrease with disease progression.The HIV-I populations of plasma and PBMC may corne from different origins.
S. Kato, 35 Shinanornarhi, Shinjukuku, Tokyo, 160 Japan Telephone: [phone] ext. 2692
Fax: [phone]
Tu.A.2070
MONOTYPIC HIV- I SUBTYPE B IN GUADALAJARA, MEXICO.
Vazquez-Valls E*, Cheingsong Popov R*., Sierra-Quevedo J.J*, Lister S*., Lopez Marquez
F*., Weber J., Zuniga Gonzalez L*., Campos-Lopez P'., Esparza J**". 'Centro de
Investigacion Biomedica de Occidente, IMSS, Guadalajara, Mexico. '*St. Mary's Hospital
Medical School, London U.K. '**UNAIDS, Geneve, Switzerland.
Objective: To identify HIV- I subtypes circulating in the west of Mexico.
Methods: Thirty HI\/ I infected subjects were enrolled in this study in Guadalajara, Mexico
between 1994-95 with full consent. A confidential questionnaire was given to the studied
subjects in order to obtain epidemiological data on sex, age, sexual preferences, and risk
factors to HIV- I infection. Serum samples and peripheral blood mononuclear cells were
collected.The serum samples were tested for HIV- I antibody (Abbott) and confirmed by
Western Blot (Organon). HIV- I viral subtypes indentified by V3 peptide ELISA and by het
eroduplex mobility assay (HMA) were analyzed with the epidemiological data.
Results:The majority of the subjects were infected with HIV-I subtype B (f- 13, m - 17)
regardless of their sex and risk factors.These were 24 subjects who acquired HIV- I infection by sexual transmission and 5 through blood transfusion, and I by paediatric HIV I
infected case Among the male subjects (n- 17) there were 10 homosexual, 6 heterosexual
and I bisexual nuen. Six male subjects have had sexual contacts in the US, Europe, Central
and South America.
Conclusions: I-IV I subtype B is prevalent in Guadalajara, Mexico.There are no differences
in the frequency of the B subtype in the studied subjects in terms of their sex, sexual prac
tices, and the routes of transmission.This report indicates that HIV I subtype B ican be
transmitted among heterosexual men and women as well as in homosexual rmen and via
blood transfusion.The biological properties of the B subtype in terms of cell ttropism in
relation to transmission needs further investigation.
Dr. Eduardo Vazquez Valls.Apdo. Postal 2 227 Guadalajara, Jalisco 04428 I. exico. Phone
(523) 6189410 fax [phone]
Tu.A.207 I
GENOTYPING OF MEXICAN HIV- I ISOLATES.
Gudino ose (armen, Martinez Fernanda, Alcantara Patricia and Soler Carmen. Unidad de
Investigacion en Retrovirus Hfumanos. Istituto de Investigaciones Biomeclicas, UNAM and
Instituto Nactional de Diagnostico y Referencia Epidemiologicos, SSA, Mexico
Objectives.To define the genotype of Mexican HIV I isolated during 89 93.
Methods. Nested PCR arnmplification reactions were carried out with the Ieteroduplex
Mobility Analysis HIV- I env Subtyping Kit provided by the NIH AIDS Research and
Reference Reagent Program.Three different DNA firagments (1,.2 kb; 0.7 kb and 0.5 kb)
were amplified isolates from hemophilic and blood transfusion and sexually and vertically
infected patients were used for DNA extractions, as well as maintained in cell cultures.
Twenty one reference plasmids belonging to the 8 different genotypes reported to date,
provided with the kit, were used as reference strains.The results are reported as the ratio
between the distance from the well bottom to the midpoint of the heteroduplex bands
and the midpoint of the hIomoduplex bands.
Results. Our- results show that the Mexican HIV- I isolates of 89-93 origin all belong unanmbiguously to the B genotype by -MA typing with the 3 env regions tested (VI toVS,V3 to
V5 and V2 to V4) Blood transfusion and hemophilic related viruses as opposed to sexually
and vertically transmited isolates show discrete and clear heteroduplex bands with the C
subtype viruses, although their ratio is smaller than with the B viruses showing a greater
genetic distance with them.This observation is particularly important for the V2 V4 frag
moent.
Conclusions. Our results corroborate the epidemiological data about the origin of the epidemic in Mexico as belonging to the Western type of transmission with the B subtype.
I omeet drue to our greait interchange 'with Central Arnetrca through our Sorrthernm borde,
monitoring of the appearance of other subtypes becones a necessity
J.C. Gudino, Carpio 470, Santo Tomas. CP II 340. Mexico D.E Mexico Tel: 34 I4 106,
3414700, 3414820. Fax: [phone] E-mail: [email protected]
Tu.A.2072
SEQUENCE ANALYSIS AND BIOLOGICAL CHARACTERIZATION OF HIV - I
ISOLATES FROM ROMANIAN ORPHANS AND ADULTS
Holm - Hanson Carol*, Rustad S*, Pascu R", Negut E'*, Asjd B'. 'Universit, of Bergen,
Norway; '0University of Medicine and Pharmacy Tirgu Mures, Romania; 'Cantacuzion
Institute, Bucharest, Romania.
Objective:To characterize HIV I isolates obtained from orphans in Romtnit 2 yers after
the 1993 baseline study and to identif, HIV I subtypes in Romanian adults
Methods: Peripheral blood mononuclear cells (PBMC) were separated fnom ED FA treated
whole blood within 48 hours after collection from 29 surviving orphans included in the
I993 baseline stud, and 9 IV - I seropositive Romania adults. CD4+/CD8+ T lymphocyte

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