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

Tuesday, July 9, 1996

Tu.A.144 - Tu.A.152

Conclusions: Different HIV subtypes appear to differ in their ability to infet and replicate in
HBCE.The examination of HBCE anrid their interaction with other primary cells and various
HIV subtypes will provide explanations of how these interaction s may allow virus to cross
or affect the blood-brain barrier and potentiate the onset of ADCa i a.
Dr. Jens Oltrogge, Georg-Speyer-Haus, Paul Ehrlich-Str. 42-44, 60594 I a,1 i 1 r a, many.
Tel: [phone], Fax: [phone].
Tu.A. 144
INTERACTION OF HIV AND SIV ENVELOPE GLYCOPROTEINS WITH LIPIDS AND
GLYCOLIPIDS
E. BahraouiI, M. Moulard2, R. Elmeskini2, S. Canarelli2, And L. Montagr ie I 1.1 UlS IToulouse
France- 2: Universitd Aisx-Marseille II France - 3:Institut Pasteur France
Objective:To study the interaction between HIV -1I, H-IV2 and SIV enveloppe gilycoproteins
and glycolipids.
Methods:Two comnplementary approaches have been used to investi7.rte ther it.e action
between HIV- I, HIV 2 and SIV envelope glycoproteins and glycolipids (gailactosy ceramride,
glucosylceramrcide, lactosylceramide) or ceramide. In the first approach, glyclipids were
chromatographed on HPTLC or fixed on microtitratiorn plates and then incubated with
radiolabelled glycoproteins.The second approach used a plasma rrmeibrane model of
monomolecular films composed of ceramide and different glycolipids.
Results:The analysis of viral glycoproteins and glycolipids according the first approach,
showed an interaction between HIV or SIV envelope glycoproteins rand v alctosylceramide,
as described previously by others, but there was also an interaction with glucosylcerarmide
and lactosylcerarmide. More surprisingly this approach enabled us to demonstrate an interaction between glycoproteins and ceramide. In order to confirm these resuits,, second
assay involving a plasma model composed of ceramide and different glycoproteins was used
to study this interaction. Here again, it was found that in addition to the interaction of
recombinant HIV I glycoproteins gp160/gpl20 with the glycolipids (galactosylceramide, glucosylceramide and lactosylceramide), these glycoproteins could also interact with ceramide.
Conclusions: In summary our work, using different techniques, has showrmn thiat the binding of
HIV and SIV glycoproteins to GalCer, GlucCer, LacCer and ceranmide is dose dependent, can
be saturated and can be selectively inhibited by envelope glycoprotein antibodies. Ihese find
ings enable us to propose hypotheses to understand the infection of C 4negative and
GalCer-negative cells, as has recently be described for primary hurnan brain endoathelial cells.
Bahraoui Elmostafa - Universite Paul Sabatier-- Bat. 4R3 I 8, Route de Narbonne - 31062
Toulouse Cedex
Tu.A. 145
INFECTIVITY OF DIFFERENT HIV-I STRAINS IN THE HUMAN PLACENTA IN VITRO.
Polliotti Brun M.a, Demeter L.b, Subbarao S.c, Keesling S.d, lee G.R.d, Caba J.d, Panigel M.d,
Nahmias A.].u, Reichman R.b, Miller R.K.a. Depts of Obstet/Gyneco la, Env Med and Medb,
and University of Rochester; NY; NCID, CDCc and Dept of Ped Inf [Dis, rEmory
Universityd, Atlanta, GA.
Objective:The aim of this study is to investigate the infectivity of HIV I strains i; the
human placenta in vitro.
Methods: Three viral strains are utilized: two laboratory strains: Bal arid llb and VI 5, isolated
firom an HIV infected baby at birth (viral culture and PCR positive). Bal a dI VI-5 are nonsyncytium-inducing (non SI) as assessed in MT2 cells and both are rmacrophage tropic. IlIlb is
syncytium-inducing (SI) as assessed in MT2 cells and is lymphocytetc pic. Human placentae
from first and third trimesters are incubated for 24 hours with free virus from each strain
before an intensive rinse to remove the inoculum. Placental villi are cultured for 5 days post
inoculation, Infection is documented by p24 release into the culture medium and by viral
DNA detection via PCR - gag gene using two primer pairs (JM850/JM85 I and
JM852/JM853) after extraction from the incubated tissue.
Results: Only the two non-SI strains (Bal and VI-5) are able to infect placental tissue both
from first trimester and full term: p24 production increases during the third day post inoculation and remains sustained for the next three days; PCR detectionr of g g gene is consistently positive at the end of the incubation. P24 release increase can be completely abolished by the use of HIV-Ig (I/100 for one hour exposure before inoculation).The SI strain
(IlIb) cannot infect placental tissue: p24 remains at background levels and PCR detection,
like non-infected control, is negative.
Conclusion: We can infect first and third trimester human placental tissue with different
HIV I strains. In addition, non-SI viral strains may be more efficient in infecting i vit ro the
human placenta, which has been observed for strains that infect the baby i, utero.
(NIH grants # AI- 32319 anrd Al 3234 I)
Bruno M. Polliotti, Department of Obst/Gynec, Univ. of Rochester Medical Center; School of
Medicine & Dentistry 601 Elmwood Avenue Box, NY I 4642-8668. USA.
Tel: [phone]; Fax: [phone]; Email: [email protected] 1IESFER.EDU
Tu.A. 146
KINETICS OF HIV-I TRANSCRIPTION IN CELLS OF LYMPHOCYTES AND
MONOCYTE-MACROPHAGE LINEAGE
Vellani N. N.,* Mo, T,* Leung, B.,* Cassol, S.* *B.C. Centre for Excllece in HIV//\IDS, St
Pauls Hospital.Vancouve, B.C.
Objective: lo study the Kinetics of HIV- I transcription in the cells of m acropirage rronocyte
lineage and lymphocytes.
Method: GEM (lymphocytic) and U937 (promonocytic) cells infecte with AVI were har
vested from O to 20 hours post infection (hpi).Tat, rev and nef transcripts we-e quantified
as copy numbers by the method of RT Quantitative-Competitive PR. Ihis method included cDNA synthesis, amplification in the presence of a competitive tempiate, ard separation
and quantinfcation of the fluorescent PCR products by an Automated Sequencer (ABI).
Results: The transcription pattern of LAV I in GEM cells was that of a n rive infection and
in U937 cells was that of a latent-like infection. In both cell lines, n re uatory transcripts,
tat, rev and nef were detectable as early as 4 hpi, although the copy a':umbers of tat and rev
were considerably higher in GEM than in U937 cells. In U937, du ri tire early times, the
number of transcript copies of nef ranked the highest and tat raled the lowest. However
is the time progressed, the number of nef transcripts declined and tIe number of tat transcripts increased.

Conclusion: ilt and nef transcription displayed opposite Kinetic trends in U937, contrary to
that seen in CEM.The predominant expression of nef and/or low expression of tat during
the early timres in U937 cells may be involved i promoting the latent like state. Cells of the
macrophage -nmorocyte lineage and lymphocytes are itajor reservoir in infected individuals.
Therefore, sttdy ng HIV- I eplication i these cell-types woul d be of cnsiderab le brenefit as
it may give rs ur better tinder standing of HIV replicitio arid its asiarairtior to te pr ocess
of AIDS.
Nina N.Vellani. RRin. 6 I 3, 1081 Burn and st.Vancouver. B.(.,V67 I Y6
Tel: [phone] 5615 email: cass lIthivn ubc. a
Tu.A. 150
SENSITIVE DETECTION OF HIV-I DNA BY ONE-TUBE SEMINESTED PCR
COMBINED WITH SOLUTION HYBRIDIZATION-EIA
Awatch ia r garn u PKunakorrn r orogkol, Raksakait K. Petcclai B. Clinrical Irnmmurnology IaIbo atory,
Depar tment of P tology Faculty of Medicine Ranmath ibodi Hospit al, Mahidol University,
Bangkokl f hailand. ThIs work is su(sllaorted by thie jpfrnese Foundstirn for AIDS Prevention
Objective: Single round polymerase chain reaction (PCR) followed by menmbrane hybridization, or nested PCR without hybridization are the usual stra tegies used for detection of
IIIV I DNA in peripheral blood cells. However, membrane hybridization is laborious and
time consuminy while contamination remains the major problem of nested PCR. In this presentationr, we showed the development of one tribe seniinested PCR (()SPCR) combnined
w Irt soluior Iylridizatiot n enzyne inm sunoass;,ry (SI IIA) whic ch i, ahieve hir t s sensitivity
and avoid thie problems previously mentioned.
Method: A low annealing temperature nested primer were used altogether with the high
anntealngh temnperature outer primners in OSPCR which was successively run for 60 cycles.
1 he PdR products was hybridized with digoxigenin probe in liquid phase, then captured by
streptavidin microtitrationr plate. via the biotin icroa p labeled onto one of the pr irwie, and
detected by clir irmetric method.
Result: Conpased at same nurnber of cycles, )SPCR has a higher sensitivity than the ordinary PCR In addition, SI HEA showed quant ttiave optical density values in detection of
OSPCR product s from the template ranges of 5 1000 HIV I DNA copies.
Conclusion: [he OSPCR conmbined with SI El/\ provides a sensitive arind rapid method for
dete, tior or HIV I DNA.
Mongkol Kunako rn, Dept. of Pathology Ramrathibodi Hosp., 270 R aman VI Road, Bklk 10400,
I hailand.-Eel 66 2011 I 1379, Fax [phone], email: rarsknsrahidol.ac.th
Tu.A.15 I
MULTI-CENTER EVALUATION OF AN HIV-1/2 RAPID ASSAY IN 9 COUNTRIES AND
USING SEROCONVERSION PANELS
C onstantine N I Mohamed Abdel-Hamid, Zhang X, San gare A, HIolm -ansen 2.
University of Mlaryland School of Medicine, Baltimnnore, MD, USA; I Institute Pasteur;s Abidjan.
Cote d'Ivoire, '-University of Bergen, Bergen, Norway.
Objective: io evaluate the accuracy of a new I IV 1/2 rapid assay with sera fiom 9 coun
tries, and using 4 HIV seroconversion panels.
Methods: A total of 1,472 sera were included and originated fron:Ta nzania (I 7 1), Cote
d'Ivoire (20), Per u (283), Egypt (200), Uganda (I 07),Vietnam (10), Indonesia (43), Ecuador
(8), and the U.S. (600).The total number of positives were 339, and included 16 HIV-2, and
4 HIV 1/2 dual reactors. Testing was pearformed using the RED-DOT HIV I &2 rapid assay
(Catalina, USA) and results were compared to those of routine ELISAs anid Western blots.
Also, the ability of the rapid assay to detect early infection was assessed using 48 samples
from 4 HIIV seroconversion panels (Serologicals, USA).
Results: All confirmed positive samples, including the HIV 2 positives were detected by the
RED DOT I /IV I &2, yielding a sensitivity of I00%.The number of false positive results
ranged from 0-5 per study, resulting in specificities of 97.7-100%.The overall specificity was
99% (I1I 33/ I 144). When testing the seroconversion panels, the RED DOT HIV- I &2 assay
detected infection at the same time, or earlier, than four ELISAs and a Western blot.
Conclusions: In this multi-center evaluation, the RED-DOTF HIV I &2 rapid assay exhibited
a 100% sensitivity and an acceptable specificity. It is capable of detecting HIV-2 positive sarnples, and is equivalent to other screening assays for detecting early HIV infection.
Mohanmed AbdelI-Harnid, 758 MSTF, 10 S. Pine St., Baltimnnore, MD, USA.
Telephone: [phone], Fax: [phone]
Tu.A.152
QUANTITATIVE DETECTION OF HEPATITIS C VIRUS AND HUMAN
IMMUNODEFICIENCY VIRUS RNA IN BRAZILIAN COINFECTED PATIENTS.
L.L. Lewis-Ximenez, M. Schechter2, H. Schatzinayr1, C-r.TYoshida,TC. Quinn3. I National
Reference Ctr for Viral Hepatitis, FIOCRUZ Rio de Janeiro,;2Univ. Hospital Clemnentino
Fraga Filho, Federal Univ. of Rio de Janeiro; aNIAID Bethesda, MD, U.S.A.
Objective: To determine the effect of HCV and HIV coinfection on viral load of each viral
infection cmpared to individuals infected with only one in fection. and to investigateI teeir
possible inerractions.
Materials and Methods: Sera fIor three groups of idividuals were divided according to
their HlV/IlV status: group I: / I IHCV+/HIV sera; yroup 2: 58 HCfV+/HIV+ sera; group 3:
69 HCV-/HIV+ sera. Quantitative levels of HCV and HIV RNA were assessed by combined
R FICR sAmplicor HCV & HIV Monitor Roche, USA).
Results: Group I: HCV RNA was detectable in 52 (73%) of 71 with a rediau of 14,372
copiesil (ranie < 100 - 1,005,556). 48 patients (68%) had previous history o transfusion
w th a median of 13,323 copies/ml, while nontransfused group has 51,449 copies/ml.
(p0.7, NS). Group 2: HCV RNA was detected in 37 (64%) and HIV RNA in 57 (98%) of
58. Overall redirn viral levels for HCV and HflV RNA were 34,598 copies/mI (rnuge <
100 3,/1,207 copies/mil) and 16,401 copies/mI (<I00 - 891,000 copies/rl), respectively.
Sex al trTiSsion was observed in 38 (65%), parenteral transmission in II (19%), both
se-ua and parenteral in 6 (I0%) and 4 (5%) with unnown modes of transmission. HCV
lNA median levels were respect vely: 12, 1-1 copies/nl, 75,204 copies/ml, 146,653
copies/ml and 0 copies/ml. Both HCV and HIV RNA levels inreased in patients with
decreasing CD4 counts. Group 3: HIV RNA was detected in 15 (94%) of 69 t atents with
a medunr of 12,000 copies/ir (r me < 100 l7,000 ies/rim)

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