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

Tu.A.105 -Tu.A.143

Tuesday, July 9, 1996

Tu.A. 105
V3-BASED PEIA FOR SERODIFFERENTIATION OF HIV TYPES AND SUBTYPES: NON-B
HIV-I SUBTYPES CONTRIBUTE TO THE EPIDEMIC IN GERMANY, SPAIN AND ISRAEL
Dietrich, Ur sula: Ruppach H, Gehring 5*, MaayanS 5+, Soriano V#, Kneclen 1$, Jaeger -!,
Ruebsanirmen Wairmansn. (I) Georg Speyer -HHaus,FrankfurlGermanmy; + adassah
Medical University, Jerusalern,srael; #Instituto de Salud Carlos IIl,Madrid,Spain; $Praxen-zentrrn BlondelstnAacien,(Gerrnanry; ~Kurtonum furr Immunschwaeche,Muenchen.Germany:
BAYER AG Wuppertal, Germany
Objective:To provide practical methods in order to monitor the distribution of HIV types
and ubt'ypo n different countries and to be able to follow changes in the HIV epidernmic.
Methods: AV3 based PEIA system was developed and evaluated for efficient serodifferentiato of HIV' I arind HIV-2 as well as several HIV I subtypes.The seroreactivity of about 700
HIV positive sera was evaluated, originating from Germany Spain, India, Rwanda, Brazil and
Th, land and representing HIV I subtypes A, B, C, E and F as well is HIV2. In order to cor
relate sr ological reaction patterns in the PEIA with the corresponding HIV genotypes,
either lare inurnber of sera with already known genotype was tested or genetic analysis
of time V3 region was performed subsequently by direct sequencing. Fhe feasibility of the
system was demonstrated testing 150 sera from Israel, a country where nothing was known
so fr -about the HIV types and subtype, present.
Results: Differentiation between HIflV I and HIV 2 as well as between HIV I subtypes A,
B, C and E crn be achieved at the sbased on the seroreactivity pattern with four different
V3 peptides. Ambigous ori absent seroreactivity patterns are mainly due to the presence
Of subftypes olther than A, B, C or E, to single amino acid exchanges in the V3-loop or to
low antibody titers in seroconvertors. Based on the V3-PEIA reactivity we could detect
Imultiple HIV- I subtypes in Israel. Whereas Israelis and Palestinians were infected by HIVI B, he large Ethiopian population ws infected by HIV I C. Occasionally, we also found
HIV IA and D Land an intersubtype recombinant. Genetic analysis was performed on 23
samiles from Israel representing characteristic or abnormral reactivity patterns in the
PEIA, different risk groups or foreign origin of Infection. In only one sample, serotype and
genotype did not rmatcIh. In additiorn,, we used the V3-PEIA to classify recent HIV- I infection in Germanyi according to viral subtype. Among II HIVI infections acquired by
Ger n-ans in the last 3 to 12 months, we found 3 times subtype E, I time subtype C and
7 t ines subtype B.Thus, non- B HIV I subtypes clearly contribute to the Western HIV
epidemic amonyg t he new infections.
Conclusion: This V3 PEIA allows to differentiate between HIV I iand HIVi2, as well as
betwveen -HIV- I subtypes A, B, C and E based on the seroreactivity pattern with 4 peptides.
Therefore, the ystve r y s very suited to monitor the HIV epidemic in large populations in
teris of the prvalence of these HIV types and subtypes. New features of the epidemic
like,ntering of new viral subtypes into a population can be recognized as well as infections
with more than one subtype. (I) and the WHO Network for HIV Isolation and
Cha, icteriztion, Geneva, Switzerland
U. Dietrich, Georg Speyer Haus, Paul -Ehrlch- Str: 42-44, 60596 Frankfurt, Germany
Tele:one: 49- 69-63395 21 6 Fax: [phone] email: brieser(a)em.uni-frankfurt.de
Tu.A. 140
HUMAN HEMATOPOIETIC PROGENITOR CELLS BECOME SUSCEPTIBLE TO
HUMAN IMMUNODEFICIENCY VIRUS TYPE-I (HIV-I) INFECTION AFTER
EXPOSURE TO HUMAN HERPESVIRUS 6 (HHV-6).
Fur liri G', RE MC*,Vigsnoli M, Ranirazzotti E*, La Placa M' *Institute of Microbiology of the
Un sersity of Bologna. 9,Via Massrenti. I 0138 Bologsna, Italy).
Objective: To study the effect of concurrent HHV-6 infection on cell sufice markers and
susceptibility to HIV I infection of two different herrmatopoietic progenitor cell lines:TFand KG-IA, and normal bone marrow human hernatopoietic (CD34 +) progenitor cells.
Methods: TF I (100% C)D31-positive, erythromyeloid progenitors which express CD4
molecules at a very low level) and KG- IA (100% CD34-positive, 100% CD4-negative, lynmphomyeloid progenitors),is well as samples of normal bone marrow human hematopoletic (CD3 4 +-) progenitor cells purified using anti- CD34 MoAb -coated magnetic beads, were
cocultivated with HHV-6 -infected JJahn cells in Transwell Col plastic plates, separated by a
porous membrane. Two days before and after HHV6 contact, cells were challenged with
HIV I (strains 111B andc Bis). Culture supernatants and cells samples were collected every
'othery 1dfor two weeks,and tsteed for the presence of HIVI protein (p24 antigen detecs,ssay) and irtegrated HI/ i [DNAorr unintegrated or cell- free HIV I RNA by PCR
and PT PCR respectively. A number of surface markers (integrins and virus related molecule,) were an,zed by cytofluorinet-ic analysis before and after contact with either
HIV I and HHV 6
Results: Both cell line s and purified C(D34+ bone marrow cells were not susceptible to
HIV I infection. In the presence of a concurrent HHV 6 infection, both cell lines became
susceptible to HI1V I infection and per-rmissive to HIV I replication, lthough with a hmited
virus yield. IF-I cells did not show any consistent modification in the cell surface markers
tested, while at least 50% of KG- I cells became CD4 -positive (antigen and mRNA). Normal
bon marrow CLD34+ cel were not always susceptible to HHV6 infection. In fact only
two olof seve n samples were found IHHf/ 6 postive and only those showed signs of
HIVI rephie Ion
Conclusions: TI ese results, wh i suggest a further possible mechanism leading to penpher
al bood d,.topenis n HIIV I rfnected subects, ccay help to ciarify tle controversial issue on
the,usceptiblity f humnr hentopoietic progenitor cells to HIV I infection.
G. Fuirs, 9 Via I-assarenti, 1icrobioloysia St. Orsola Gen. Hospital, 40 I 38 Bolorna, Italy
Telephone: + 39 51 34i652 Fax: A 39 5I-341632 email: got [email protected]
Tu.A. 141
HIV AND SIV DOWN-REGULATE THEIR OWN INFECTIVITY POTENTIAL
THROUGH VIRION SURFACE SIALYLATION
NtaiY, Hu H, Moriya C, Xin X, Hasan M, Shioda T Institute of Medical Science, University
of Tokyo,-Tokyo, Japan
Objective: To learn the signmcance of heavy glycosylation and sialylation of virion surface for
the.nfectivity and cytopathogenicity of IIV I and other human and non human lentiviruses.
Methods: T cell line tropic HIV I strains SF2 and NL43 were treated in vitro or cm culture
with neuramin dase and their replication kinetics and cytopathogenicity were examined.

Effect of ir1bo - r. n m iriosidase I (deoxymannojirimnycin) and mannosidase II (swanso
nine) on t e, -, n was also examined. Similar studies were extended to the
macropha 'e p 62 and primary isolates of HIV I as well as HIV2 strain
GH123adi', - 2'39.
Results: Eri, -,,-,ion of HIV- I strain NL43 and SF2 imm-ediately resulted in aug
mentation of i. t,--ction with host cells in a fashion hlikely undescnbed before and
lec to remark ' Len r, ed viral replication and cytopathogenicity.Well complemsent ng
results camnoe frri stde w ith the inhibitors of mannosidase I and mannosidase II, demonstrmting that these inhLtors cause strikingly similar enhancement of HIV I relication and
ctopathogencity as found with neurami;idase. iEnharncement of infection by neurm ini idase
was also found with HIVI strain SF162 and primary isolates as well as HIV-2 iand SiV.
Conclusions: The human and non-hurnan primate lentiviruses appear to down-regulate
their own infectivity potential through the virion surface sialylation, thereby determinan;
their individual ordinary infection efficiency Full processing of the viral olgosaccharde chains
into the complex type appears to be impedimental, but neither- necessary nor indifferent, to
the expression of full infetivity
Y Nagai, 4-6- I ShirokunedaM, Minato-ku,Tokyo 108, Japan Telephone: [phone]
Fax: [phone] email: ynagal(ihg. ms u-tokyo. ac. ijp
Tu.A. 142
HIV-I REPLICATION IN EPITHELIAL INTESTINAL CELLS
Chenine Agnmes--Laurence, Sanchez G, LeContel C, Godard C, Chermann J-C, Hirsch I.
INSERM U322, Unit de Recherches sur les Retrovirus et Maladies Assocees, B.P 33. I 373
Marseille Cedex 09, FRANCE
Objective: The human colon epithelial line HT29 was used is a model for study of persistent and abortive infection with HIV I in CD4 negative cells.
Methods: infection of HT29 cells with different subtypes of HIV was followed by produc
tion of reverse tr anscriptase activity or p24g to cell free supernatant or by cocul,tlon
with MT4 lymphocytes.
Results: At least thousand HT29 cells had to be rinfected with HIV- NDK. a chde D
Za srian virus highly cytopathic for CD4 positive lymphocytes, to establish a massive and persistent virus production in a total population of 106 intestinal cells. Below this eritical pro
po tion, the HIV-I NDK infected cells were progressively aborted from the culture, like ir
the case of the prototype clide B virus HIV I LAV Infection with primary clinical solates of
subtypes A, B, C, D, E and F was also abortive. Low production (<_102 TCIU/ml), charactenstic for a period of several weeks after infection of HT29 cells with HIV I NDK produced n
CEM lymphocytes (HIV I NDKCEM), was followed by an adaptation and massive virus production (I 06 TCIU/mi).The adapted HIVI NDKHT29 virus was about hundred times and
ten thousands times more infectious for HT29 cells than HIV- I NDKCEM and HIV- I LAV
respectively After adaptation, HIV I NDKH-T29 was produced and its DNA was detected
about 10% of HT29 cells whereas the rest of cells was free of the virus or its DNA. HIV
NDKHT29 producing cel! clones derived from persistently nfected culture divided about
50% slowlier th in non- infected cells. Infection of intestinal cells with HIV NDK is not followed by cytopath ic changes visible by optical microscopy Their cocultivation with
macrophages, but not with lymphocytes, infected with macrophage -tropic virus HIV- I PAR
resulted in drastic changes in their growth properties and morphology
Conclusions: Our results indicate that difference between persstent and abortive infection
depends on dynamic equilibrium of (i) kinetics of HIV Iproduction. (i) its infectivityfor
intestinal cellsI. i) reduced rowth rate of infected cells related to their negative selection
and (iv) overgrowth by uninfected cells
A.L. Chenine, INSERM Unite 322. B.P 33, 13273 Marselle Cedex 09, France.
Teephone: (3) 9 I82 75 40 - Fax [phone]
Tu.A. 143
CAPILLARY ENDOTHELIAL CELLS DERIVED FROM HUMAN BRAIN TUMORS ARE
PERMISSIVE FOR HIV-INFECTION.
OItrogge, J.", H. von Briesen*, B. Engelhardt*", C. Pereda-Fernandez*, U. Woelk ",W
Schlote*', R. Lorenz'Y, H. Rubsamen-Waigmannit**, R. E. Unger*. Chemotherapeutical
Research Institute, Georg Speyer Haus, Frankfurt, University of Frankfurt" *. Max PlanckInstitut for Physiology and Clinical Research, Bad Nauheir'"*, and Baye. AG.
Wuppertal"* "', GERMANY
Objective: Infection with the human immunodeficiency virus (HIV) not only results,
inmmune system dysfunction (AIDS) but also frequently in u denenting neurological disor-der
of the brain, or AIDS demnentia complex (ADC), which can be clinically distinguished from
other CNS complications due to AID)S. Individuals with ADC exhibit abnormalities in motor,
cognitive and behavioral functions, which become progressively worse as the health status
declines.Virus has been detected within macrophage, glial and neuronal cells in the brain,
and ADC is beheved to be a consequence of infection of these cells. How HIV crosses the
bIcod -brain barrier made up of tightly packed brain cap illary endothel l cells, which is normally impermeaoe to other cells, bacteria, viruses or sntibod ies, to reach these target cells
within the brain is not knowThe objectrve was to solate human br-ain caplars endothelial
ells (HBCE) and to determine their infectivity with different HI ssubtypes and the conse
quences of HIV infoctIon.
Methods: Opt mal conditiors were established for the r-ecover isolation and mintenance
of human brain capillary endothelii cells (HBCE) fromn freshly removed brain sunor- c
as meningiomas, gioblastomas, adenocarsnomas n tissue culture systems from HIV
se onegative adults. A number of different freshly solated HIV subtypes as well as lab straun
of HIV which exhibed different ir vitro cell tropisms were used for infectity studies and
the effects on ctopathology of IBCE, replication of vrus, ard interacton with other cells
were examined.
Results: Endothelial cells were isolated from cells that grew out of bran caplary segrcents
recovered by enzycatc dgestions and gradient centrnfugitions of homogen zed bra n
tumors. Cultures were greater than 95% endothelial cells after the second passage mnd
could be maintained for at least 10 passages.These cells formed simlar tight junctions as
s-n rive, which are responsible for the impermeabi ity of the blood-bran barrier and
exhibited typical endothelial von Willebrand factor staining and binding o UEA-I
Preiminary HIVinfectivity experiments rndicated that nfection of HBCE occurred but was
dependent on viral subtype and the method of propagation of vra stocks.

217