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

Tu.A.2095 - Tu.A.2100

Tuesday July 9, 1996

in that some licenrsed HIV antibody screening assays failed to detect some individuals infected with this strain. Most infections with HIV- I' group O strains have been reported among
people fronm- Cameroonr, (aon and Nigeria, some of them living in Belgium, France, and
Germany.
Patients and methods Because a high proportion of patients attending our institution are
immigr, ints, namely from West Africa, al sera tested for HIV were firstly analyzed using a
mixed antigen HIV I plus HIV 2 IA, and repeatedly reactive specimens are further con
firmed using both an HIVI vii lysate Western blot (VWB) and osynthetic peptide- based
assay (Pept-LAV 1+2). In this form, HIV2 antibodies are more definitively excluded than
excus vely using the recommended HIV I WB analysis. Given that mostly HIV - I roup (.)
sera are negative on Pepti- LAV after being EiA reactive or cut off, the mentioned algonrtm
provides us the opportunity to collect a group of that samples might be candidates for HIVI group O infection. Specimens selected by this serological critenra and additional sera from
HIV I + subjects from West Africa (epideririgcol crteria) were analyzed using a new line
immunoassay from Innogenetics (Ito LIA HIV type O) which can recognize antibodies
specifi for HIV type O.
Results: Since January 1993, 832 sera were found to be reactive by one HIV I/2 EIA.WB
and Peptl LAV confirmed 665 of them ais HIVseropos. None of 23 samples firom HIV I +
Africans yiel ded reactivity on the no-IA HIV type O. Howeve one sample reacted on
this test after testing 77 set a which yieded reactive or cut-off absorbances on EIA, Indetermlnate WB patterns and negattve resu lts on PL.The sera conrresponded to a 34-year old
wom born in Spain, which had a monogamous sexua relation for the ast 6 years with
35 -eor-oldir, lie, and previusly only spurd ialiy with two other persons, all of whom
denied risk pract ces. Howe ver he curenrt couple had been on duty n EquatorialGuinea
for 10 years (1983-1992), where he had had mutiple sex partners Besides, he had ti aveled
several times to Cameroon.The husband was EIA-reactive aond showed posit-ity for HIVI
on W and P-L. Iiowever ie orly ected against HIV group O d not r ant ed on the
Inno-L tA HIV,group O.
We uoed prewousl ides ibed nested primer pudrs fro the e region for differentiatron
between group and O cvruses n to wpatients. Both yielded posictve results exclusely for HIV group. Moreover; seq, ence analysis of e ov and f renomic regions confirmed
these results and allowed ui to cluster our cases into the ANT-70 lade nstead o the
MVP B 180orVAU.
Conclusion: The reported cases of HIV-I,roup 0 infection in two persons of Spanish ori
gin rase the possibility of the spread of dcerent HIV I isolates wordwide. In addition, it
reinforces the utility of periodic surveillance studies at the international level in order to
adapt current screennp oserological tests to the evolvin HIV epidemic.
Dr.Vincent Sorano. Service Infectious Diseases, ISC III, C/Rafrel Calvo 7, 2 A, 28010 -Madrid, Spain. Phone: (34 I) 3 140807. Fax: [phone]
Tu.A.2095
MOLECULAR INVESTIGATION OF HIV-1 VARIANTS IN UKRAINE
Grebenjuk Vladslv A', Anoprienko O ' Marichec I*', Kavsan V'. Institute of Moecular
Biolog and Genetics, Klev, Ukrine: Institute of Epidemiology & Infection Diseases, Kiev
Ukraine.
Objective: 1o determine the extent of HIV- strains heterogeneity in Ukraine.
Methods: Blood samples were collected from four (n=-4) HIVI seropositive male residents
of Ukraine.Two of them are presumed to be nfected through heterosexsual contact and
other two through homosexual contact. Al indrAiduals were first tested to be HIV. I postie
within last two years and are stil asymptomatic. DNA samples derived from peripheral
blood mononuclear cells of these pat0ents were used as templates for nested polymerise
chain reaction. The 0,58 kb fiagments of gagene encoding p17 - p24 portion of ga
polyprotein and, I kb fragmoents of env,ene encodinggpl60 C2 C5 porton were
amplified, cloned and serquenced Sequences of PCR products were aligned and phylogenetic neighbour-jorning trees eerated.
Results: All obtair ned nucleotde sequences of portions of gag gene when compared to
HIV I reference strains of different subtypes were clustered to subtype B.iThe same dstribution was found for env sequences. Both obtained env and gag sequences from two homosexually inlected patients were most closely related.This i i a good correlation with the
suggestion that they were infected by each other.
Conclusion: Thus, we found that HIVI strains derived f-om four Ukrainian residents belonp
to subtype B, which is widespread in Europe and also in Russia. Existence of any other -HIVI subtypes in Ukrainian population require further investigations.
VA Grebenjuk 50 Zabolotnogo Street. Kiev 252627, Ukraine Telephone: [phone] Fax:[phone] emal: kavsan( oimbig.kiev.ua
Tu.A.2096
CHARACTERIZATION OF HIV-I QUASISPECIES REPERTOIRE IN INFANTS AND
CHILDREN BY HETERODUPLEX GEL SHIFT ANALYSIS
Essajee, Shaffiq M. Pollack H, Rochford GC. Oransky I, Kr-asinski K. Borkowsky W. Department
of Pediatric Infectiotus Dsease, New York University Medical C renter New York, USA
Objective: Most individuals infected with HIV harbor not one but several different genotypes df the cirus. Stud es i-lult h ie estbishod a rorielaon between greater viral
dicorspty rid looper eurvah ihilr o V3 does not icst for te pedi tic population Du
m s to hiarate innz oe th on of DIV quasspecies in dnts an d children and
roetoepatters of dre no v with ucif e outcom e and parameters of rioune function.
Methods: Too subeconts 6 slow progressors nd 4 rspd propessos), were selected frorn
cohent of DIV infected chidrer followed pr ospectivelp si000 b i th.Threy had boon seen it
monthly intercils when cl ncal ecillustronr were performrod. aod DIV-I nncwtro ant body
(IVAb) production from o troed per phersl hlood lymphocytes winscmeasured. Nested DIV
DNA PCR win performertd or sequent.l specimens of peripherl ifblood mononucloar cells
to aroplify the V3-VS ropion of the onc poee Saroples wore seror-qurnified by end-point
dilutin o staindardize0copy n urobe Usrngp radio-labelied probe gonerasled irene the initial
specor en closest to Ibirth, honoroduplox gol sIt Bnlsos (GSA) was performed no yielda
piclopiraphic record fo eah child of V3-VS Bonertic vaten rover time.
Results: Ir '4 of the rnfir Itsomore thant ooe quasispOes was identlfied it hirth.The 4 rap d
propriessors showed no ecdence Iorn fus pc en ecolton duriop the period of th e study
B8-1 Bmton) By conrt, t of too p} slor pi0Bo5rs5 shtowed the appoararoo of new

quasispecies by 4-6 months In addition, none of the rapid progressors made IVAb during
the first year of life, whereas in all but one of the sow progressors, IVAb production was
detected at the time of or prior to changes in quasispecies repertoire. There was no association between the presence of more quasspecies it birth and either more rapid d esese
progression or more rapid viral evolut ion.
Conclusion: More than one quasispecies is often transm itted to verticaly infected nf nts.
For slow progressors, changes in the population of quasispeces occurred in most cases by
the fi st 6 months of life, and appeared to be associated with evidence of a strong nfant
immune response. Infants who progressed rapidlyfailed to show any changes i, t-e quasspecies repertoire and also failed to produce signficant amounts ofin- vitro ant;body in this
relatively small study w e have shown that heteroduplex analysis of viral diversity has some
predictive value in determining clinical0 outcome.The ssociation between ncreased viral
diversity and ncreased IVAb product on lends weight to the hypothesis that immune pressure plays a role in driving viral evolution. Further study is needed to shed Ight on the significance of these findings.
Shaffiq M1 Essajee, 415 East Ninth Street 91 NewYork, NY 10009 USATel: (212) 562 361 2
Fax: [phone] email:essajs0lO!gcrc.med.rnyu.edu
Tu.A.2098
NONINFECTIOUS HIV- I -LIKE PARTICLES WITH HETEROLOGOUS
IMMUNOLOGICAL MARKERS AS NOVEL AIDS VACCINE CANDIDATES
Yao Fei- LongCao S X, Cates G, JtmesO C), Chong Persson R, Matthews T, Kle n MP, Roinsk
B. Connaught Laboratories Ltd, Ontan oiCaiida.
Objective: To eng ineer anti genicall s-marked HIV ike prtIcles whi ch permi t mmunol oical
dislin ctr between vaccination and H IV-I nfecti or.
Methods: An nduoble, humosa n metalloth onein -bas eeof pr-esoruce r n sbe f I poducpn HI- like partices with chimeric gp 120MN/,p1LAI envelope lycoprote!rs wis mut- i
genized to en neer a vect orcapable of prsoducing vorus-Ike partices 0 contain- aeterologous transmembrian re anchor sequence fu sed to the carboxy-termnus of gpp120oMN. othis
end, a DNA sequence encoding a 57 me peptide contaninp 22 amino acids fom the
human influenza virus HA2 protein was inserted into specific locus wthin the HIV oenvelopeg ene by gene assembly-aided mutagenes, (GAAM).
Results: Monkey kidneyVero cells transfected with this vector were shown to produce DIVI-like prtces devoid of gp41 and contoining the modified gpl20 protein. Immunization
owith these particles induci ed HIV- type-specific net cra zins antibod ies as well as antibodies
recognizingr the heterologous angenic marker peptide.
Conclusions: These studies demonstrate the lasibn' i ity of producing noni nfectious HI - I -ke
paricles wttho pl20 envelope glycopinoteins anchored to the viralorane surace via
heteroloous antigenic peptide fragments. Since those particles doo nt contain the HIV-I
immunodominant region which is present within the tn s enrbrane glycoproten,mmunization with these HIVIlike antigens should allow a 0lewr distinction between vaccnation
and live virus nfection.
F-LYao, 1755 Steeles Ave.West. North York ON M2R 3T4 Telephone [phone] Fax:
(4 1 6) 667 2740
Tu.A.2099
MONOCLONAL ANTIBODIES AGAINST HIV- I p17 CROSS-REACT WITH ENV V3
LOOR
Ota, Akemi, Watanabe M, ShangJ, Nakanish M, Leda S. Osaka Universt Csaka, Japan
Objective: We found antibodies in serum samples of a HIV-1 carrier that cross-reacted with
envV3 loop and p17 peptides of HIV-I. Among monoclonal antbodies (mAbs) against p17,
some cross-reacted with envV3 oop.To confir rthis phenomenon, we examined newly
obtained mAbs against p17 for their cross-reactlvty with env V3 loop.
Method: MAbs aainst p 17 were obtained by immuni zin a pepti de, P30 52 (covenrng
ami no acid position between 30th and 52nd), of p 17 n Balb/c mice. Cross reactvty and
affinity constants were examined by ELISA in IBIAcore system.
Results: Among ISMAbs newly obtained, 7 were IgM type and 8 were IgG type. ELISA tests
using Covalsin k plates (Nun c) revealed that 14 of these rAbn s cross-reacted with an envV3
loop peptide. Affinity constants determined with BIAcore biosensor were between 1.4 x
10 and 7.6 x 107 (I/M'1). Indirect fluorescent antibody staining revealed that 8 mAbs reacted wth rhe cell surface of MT4 cells infected with JMH-I strain of HIV.Fou mAbs of
gM type showed neutralizing activity and 4 of I1M type suppressed multiplication of the
JM1 strain.
Conclusion: MAbs against p 17 of HIV- Icross-reacted with an env V3 loop peptde. Some
of them reacted with cel surface antigens of MT-4 cels infected with HIV I and showed
suppressive effect on HIV I multiplication.
A.Ota, 3-1 Yamada-oka, Suita city, Osaka. 565 Japan Tel:[phone] Fax:[phone]
ema l:otadobiken.osaka-u.ac.jp
Tu.A.2100
TCA3 AND INTERLEUKIN- 12 AS CYTOKINE ADJUVANTS ON CANDIDATE HIV- I
DNA VACCINE
Tesus Tadash Hamaia K. Ishir N. ADr Yan K, Saraid S. fwa rho d Fu clnoa J, Ododa
K. Yokohanta City UnroersitY hoohans sapin
Objective: To detemino whetherTCA3 and inter eudmLp 2 expression pami dn have
adf oct1 stiites or DIV-i speciic nmed 'ed immu nity p CMIw iuced 'p Ocr nddateO
DIVI DNA vacione.
Methods: TCA3 or IL- 12 expre sson plisnid was cornocuilotd roth DIVI 1118 p160 expresion planmid (hereafter DNA va50iref into cionse gastrocremtic rouncles.Two woods after
inooculat on. Delayed typo hypersenicvty (DTD roan assayed by osn 0 foolpad sine opg
nosponse.Three woods after inoculsIon, rytoocT lymphocyte CTb) acitr~teo wis 0000u
ated Op using standard 5Co~reloise is syTmo and four- weeds ifter rnoculation, the DIVspecifho antibody Inter nO rorounizod nero mrs evalcaned by using enzynto lided rmmunosor
Lent asay
Results: The DIVI specific CMI (DTD an d C TL rctortes roa errhinced by 000nocula100
of eich cytodrne oxpressron plasrord rod DNA orccno comparood itL vco no anne.TIhe
DIV-i specific honoram immunity mas rather ouppressed whenr rho vacorne mms rootled
with TCA3 exprossnor plasroidI

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