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

Tu.A.153 - Tu.A.162

Tuesday, July 9, 1996

Discussion: Viral load of HCV does differ, although not significantly, according to different
routes of transmission.The highest HCV levels were observed in the confected group who
acquired their infection parenterally compared to the single infected group. Most of the single infected group had a previous history of one transfusion while coinfected patients
icluded mostly IDUs. HCV repli cation was 2.4-fold higher in coinfected than in single
infected patients but there was no evidence that HIV replication was increased by the presence of HCVThe Amplcor f CV and IlV issays were able to detect very low titers of
RNA ( 102 103 opies/ml) and will prove useful in determining virologic responses to specific treatment inr larger studies of coinfected individuals.
L< I iura Lewis Ximenz Departan ento de Virologia, Instituto 0swaldo Cr uz - FIOCRUZ
Av. Fri il, 365, RPo de Janeiro, RJ,.Brasl-210'0- 360 Telephone: [phone]
Fax: S (1) 270 6397
Tu.A. 153
CONSISTENT SEQUENTIAL DETECTION OF RNA,ANTIGEN AND ANTIBODY IN
EARLY HIV INFECTION:ASSESMENT OF THE WINDOW PERIOD.
Bus h M., "SchumacherjRichardT., **Stmer 5., S siGarrett PE. "lrwin Memorial Blood
Center, San Francisco, CA USA, "Boston Biomedica., Inc.,W Bridgewater, MA, USA,
* *ARC, Ro(kville, MIl), USA.
Objectives: 1to evaluate the evolution of significant markers of early HIV infection and assess
their potential for reducing the HIIV window period.
Methods: Between 1987 and 1995, serial bleeds collected from individuals recently infected
with HIV (seroconversion, SC) were tested with methods designed to detect HIV antibody
(@ HlV), antigen (HIV Ag), and RNA. Stability of frozen SC samples allowed the comparison
over 8 years of detection with early @ HIV methods, so-called 3rd generation @-HIV
method,s. HIV Ag tests, and recently developed target amplfication test methods for HIV
RNA, and the concomitant reduction in time fromr infection with HIV to first marker detection (window period, WP).
Results: In 1987, the WP for e -HIV was estimated at 12 days, based on tests of transfusion- infected individuails, surveillance programs of high risk cohorts, and retrospectively recognized seroconverters from plasmapheresis programs. In 1995, a study of 31 SCs with currentl/avilible (if HIV tests (7 methods), HIV Ag tests (5 methods), and newly available HIV
RNA tests (3 methods) indicated that the WP for the more sensitive @-HIV tests was
redued to 1 8. 5 days, that screening for -HIV Ag could produce a further reduction to 8
dys, snd that screening with HIV RNA tests could reduce the WP further still, to 5.3 days.
Conclusions: I) Seroconversions consistently demonstrate the sequential appearance in
blood of I flV RNA, HIV Ag, and a HIV. 2) Detection of HIV RNA and/or HIV Ag can be
used to confirm early infection. 3) RNA and/or HIV Ag tests are potentially useful for earlier
detection of HIV nfection (e.. blood screening). 4) SCs provide a valuable benchmark for
following the improvement in sensitivity of tests for early HIV infection.
Rich<rd T Schursacher Boston Biomedica, Inc.,W Bridgewater, MA 02379
Telephone: [phone]. Fax: [phone]
Tu.A. 154
MULTIPLEX POLYMERASE CHAIN REACTION FOR DIAGNOSIS OF AIDS- RELATED
CENTRAL NERVOUS SYSTEM LYMPHOMA AND TOXOPLASMOSIS
RobertsTlC., Storch, G.A. Washington University School of Medicine, St. Louis, MO USA
Objective: Toxoplsrn encephalitis and EBVassociated lymphoma are leading causes of
space-occupying lesions of the brain in patients with AIDS. A multiplex polymerase chain
reaction (PCR) was developed for the simultaneous diagnosis of AIDS-related central nervous system (CNS) lymphoma and toxoplasmosis, and its performance was compared with
the PCR assays for Epstein-Barr virus (EBV) and Toxoplsrtn gondi carried out individually
Methods: A nested multiplex PCR assay was designed for the amplification of segments of
the I x lsirrs,ndi BI gene and the B n HI W region of the long internal direct
euere of EIBV TFhe rmultiplex PCR was per]ormed on cerebrospinal fluid samples from 52
AIDS patients with either cinically diagnosed or histologically proven CNS disease and
compared with the PcR assays for EBV and T gofdi performed individually
Results: Using the multiplex assay, Txolrsmr DNA was detected in 2 of 2 patients with
histologically proven Toxo/lasmr encephalitis, in 5 of 6 patients with a clinical diagnosis of
toxoplasmosis and in none of 44 patients without disease. EBV DNA was detected in 5 of
9 patients with proven CNS lymphoma, in 4 of 5 patients with a clinical diagnosis of lymphomai and in 2 of 38 patients without disease.These results were comparable to those
obtained when t he separate PCR assays for gondn and EBV were performerd.
Conclusion: Fhe multiplex PCR assay accurately diagnoses AIDS related CNS lymphoma
and toxoplasmoss. Given the reduction in time and costs by the simultaneous detection of
T i gonidu and EBV in one assay the multiplex PCR assay may become an important component in the evailuation of CNS mass lesions in patients with AIDS.
T.C. Roberts, Depairtment of Pediatrics, Washington University School of Medicine, One
Children's Place, St. Lour', MO 63 I 0 USA,Tel. [phone], FAX [phone],
emal: roberts Tcafl.kids.wustLedu
Tu.A. 155
HIV-I SUBTYPE SPECIFIC DIFFERENCES IN PLASMA RNA LEVELS RELATED TO RATE
AND AMOUNT OF VIRUS PRODUCED IN PBMC CULTURE
DDe f tlf rkrVon Br esNer HU oles M5 'akkor N", Cornelissen C*, Goudsmit J*.
D rmenr t of furan Retrviioley, AMh, University of Ars terdam, the Nether ands;
''freog-Slidber flaus, Fr-anskurt, Corn ny; *' NIBSC, Petters Br, DR; the 0/DO network
for-I VlY islto,irsd ch srace losit isr, cnvi, Sw Ies Iard
Objective: To rstI se whethe- differenies is rep irte o could be found between HIV-I
ifbtipes 17 c pir he eficiency of PBMC culture, as well as HIV I genornic RNA levocs ue~sed in piiisa cf rdvidus rowdy iofeted witf DIV-I subtype B, E, A or D.
Materials and Methods: IV I RNA lever were mesured by NASBA n the supereatant of
stinr sdized PBMC susr s, re 'wel as diroctly is plasews of 55 indiw duals infected with DIVsubtype B (n- I) A ( 2), D ( 3) or E (I5) smpled at a time point within 2 years of
DIV eI iboy sreccversrice
Results: Differ errs in Iise o soerrvnversicc anrd rsgo~ of DIV- infection as assessed by
moe',irnp f2 soisroy1ifusic lovels is plescca, did eel rise srn iccpact on the r-esults anid northf s'id Ih cIuso beri on lisma tches If the srice n the pli iies/procbe cogions used in the

NASBA. Infectivity of these subtypes (TCID50) was proportional to the HIV-I RNA levels
in the virus cultures. Monitoring PBMC cultures for p24 positivity revealed that subtype B
became positive after 4 and A after 8 days of culture. Subtype D (12 days) and E (I 4 days)
cultures took significantly (p=0.000I and 0.0001) longer. HIVI RNA concentrations n the
cultures of subtype E were lower and rose ait a slower rate thn in thcte cultures infected
with the three other subtypes. HIV I RNA copy numbers directly measured in plasma were
significantly lower in subtype E infected Individuals compared to ndividuals infected by sub
types A (p=0.003), B (p=0.0 I) and D (p-0.003).
Discussion: The HIV-I RNA steady state level established during the first years of infection
wilth HIV I subtype B predicts clinical outcome. HIV- I RNA level. n plasm/serum are also
predictive of the success rate of HIV-I isolason from PBMC, indicatin a link between
replicative capacty of isolates in vitro and in vivo levels of HIV IRNA. Our results suggest that
subtype-specific differences in HIV-I RNA levels rin vvo occur and are rel ated to the rIte and
amount of virus produced from peripheral blood cells of newly infected individus.
Frank de Wolf, Department of Human Retrovirology Academic Medical Center, Uni ersity
of Amsterdam, Meibergdreef 15, 1105 AZ Amsterdam, the Netherlands tel I-1 20 5665057;
fax [phone]
Tu.A. 160
ANTIBODY REACTIVITIES TO P53 AND HTLV-I REGULATORY PROTEINS-TAX,
REX AND TOF IN HTLV-I INFECTED PEOPLE
Yi-Mling A. Chen*, Shiu-Hung Chen*, Mltsuhro Osame. 'Nat onal Yang- HMng Unversity
Taipei,Taiwan; "R*Kagoshima University Japan
Objective: To understand the antibody react vities and interaction between anti pb3, antTelof anti-Rex and anti-Tax antibodies in HTLV-I -infected people.
Methods: A GSTTof expression plasmid was constructed and the fusion protein was used
for Western blot assay (WB). In addition,WBs with different antigens which nclud ing a
GST-p53 recombinant protein, aTax recombinant protein and cell lysate from HeLa cells
transfected with Rex-expressing plasmid were also developed for serological analyses.
Results: Four% (2 /50) of HAM/TSP patients and none of 50 asymptomai c carnles (AC)
or 50 ATL patients had anti-Telof antibody; while 2% of ACs, 4% of ATT L patients and 6% of
HAM/TSP patients had anti-p53 antibody The rate of anti- Rex antibody of HAM/ISP
patients (1 6%) was significantly higher than that of ACs (2%) or ATL patients (0%). Only
one HAM/TSP patient had both anti- Rex and anti-Tof antibody.
Conclusions: This is the first study demonstrating that there is anti p53, ant Rex and antTof antibodies in HTLVI infected people. Correlation analysis showed th t anti -p antbody did not have any correlation with anti iax antibody in HTLV-I-nfected people
(McNemar's test, p < 0.05). However the correlation between ant- p53 and nti-Rex anti
bodies or anti -p53 and anti Telof antibodies can not be ruled out in this study
Yi-Ming A. Chen, Nationa Yang Ming University Shih- Pai,.Taipei 1221. Tiwan;
Phone: [phone] Fax: [phone] email: ArthurOdns.ym.edu.tw
Tu.A.161
A PROSPECTIVE STUDY OF HTLV-I INFECTION
Tayor Graham P I,Tosswill JHC2, Matutes E3, Daenke S ', Bangham CRM 14, Rossor M,
Thomas D,Weber JN. St Mary's Hosp ital Medical S chool, Lon don UK; Central Public
Health Laboratory, London, UK; 3Royal Marsden Hospital, London. UK: 4 nstitute of
Mo ecular Medicne, Oxford, UK
Objectives: To determine the natural history of HTLV I infect ion in carriers, the spectrum of
subclinical abnormalities and to identify prognostic markers.
Methods: A longitudinal, prospective, clinical study of 16 HTLV I asymptomatic carriers and 4
patients with HTLV-I associated myelopathy between 199 1 and 1995. Lymphocytes were
examined in May-Grunwald-Giemsa stained peripheral blood films, Immunophenotypes
determined by FACScan, proviral load determined by nested PCR on errs dlitons of
PBMC DNA and fresh and stimulated CTL responses measured n chromnium reletse assays.
Results: We have documented episodes of uveitis,alveolitis, polymyositis and hepatitis. FThree
carriers have developed mild CNS abnormalities whilst a patient preseno1 n acutely with
HAM, progressively deteriorated over I 2 months and died. All patients and cariers have
abnormal lymphocytes (4.2% and 2.9% respectively). Flower cells are found in patients and
carriers, including those who have acquired infection in adult fe. HLA DR. CD25 and
CD38, markers of lymphocyte activation are expressed by 22.5%. 8.2% and.9% of
peripheral blood lymphocytes.The median proviral oad in PBM(s was 2.8% in the iners
and varied little with time but was I log higher in patients and carriers developing H TLVI
associated disease.The effector frequency of tx specic cytotxir T lymphocytes was simiar
in both groups.
Conclusions: HTLV-I carriage is not silent. CTL a 'vityIs the.ane in rersfnd pients
High proviral load is assocated with the development of HTLV I ssocIted dc ie ind
predates symptoms.
G.PTaylor St Mary's Hospital Medical School, Praed St. London W2 I NY UK
Tel: [phone] Fax: [phone] e-mail: GPT30(Tic.aL
Tu.A. 162
ISOLATION OF A NOVEL SIMIAN T-CELL LYMPHOTROPIC VIRUS (STLV)
FROM PAN TROGLODYTES, RELATED TO THE HUMAN T-CELL LYMPHOTROPIC
VIRUS TYPE-Il (HTLV-II)
Ya rr, R., McLed, Dan. National Laboratory forV nal Oncolopy LCDC, Health Canada,
Otte we, Canada
Objective:To irete and cha acter iz the HTLV II lae retrovrus from seropos
ci opanzee indigenous to the tropical western Aftrlirs(Gabon
Methods: HTLV-I ELISA positive chimpanzee plasma sampes were further tested by WB
sape rsing the rost advanced DTLV BLOF 2.4 k )tGeneb D nostIcs S spore) contarn ng dirtrspted DTLV-I visions sspplemeeted cct 18e combinasri?props pciic er
DTIV-I end DTLV-II. Specimens reactiso to GAG Irs19 cc p24 snd icc [NV CL]5: sead
rgp'16 I) were considered as HTLV-I positive, wh le those reacte to p24. GD21 nd r- p46 -II were consideted as DTLVII positive Coc ultures of PBMC fm si c posive ain ns wth
hunran mononuclear cells were enred out. Suscces of suna isolatccionwa mc foeted dv
postive RT detection, syncytism formatlon, RIPA, RTPCR end sod-we] shyerIclt~n

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