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

Track A: Basic Science

Mo.A.1099 - Mo.A.l 104

than uninfectrd,cells.This feature of HIV infected cells may be expl ', t!h se c f
hygromycin B r other selectively permeable compounds to treat A!0)S
Paul J. Gatti, Dept. Micro/Immunol,Tulane Medical School, 1430 Tul s i HJsew
Orleans, LA, USA Telephone:[phone] Fax:[phone]
Mo.A. 1099
PROTEOLYTIC FRAGMENTS OF ANTI-HIV PROTEINS MAP30 AND GAP3 I ARE
BIOLOGICALLY ACTIVE
Lee-Huang, Sylvia,* Huang, P Lee,*" Huang, R Lin,** Bourinbaiar, A. ' i I (
Kung, H F.* * *Biochemistry Department, NYU Medical School, N", I'. ', If l/arvard
Medical School, Boston, MA; **NICHD, NIH, Bethesda; **-*NCI-F( PIX I- erick, MD, USA
Objective: The plant sources of anti-HIV proteins MAP30 and GAP I, i;,-ern,used in
traditional medicine by oral administration.Thus the bioavailable for ' se r i iii-HIV
agents may be proteolytic fragments.The objective of this study is to deir ne whether
proteolytic fragments of MAP30 and GAP3 I are biologically active.
Methods: Proteolytic fragments were prepared by treating the anti HIV prnt'ins with proteolytic enzymes trypsin, pepsin and V8 protease.The enzymes wes r e i ved from the
reaction mixture by enyzmeinhibitor affinity columns.The fragments,were ientfied and
purified by SDS--PAGE and HPLC.The anti-HIV activity and biochenc ac wiitites of the
fragments were determined.
Results: The proteolytic fragments of anti-HIV proteins, MAP30 andii,APi retain essentially full biological activity of their parent proteins.The fragments inhibitl IVI infection and
replication to the same extent as their parent molecules as assayed in ac it y as well as
chronically infected cells by syncytial formation, viral core protein p24 expression and HIVassociated reverse transcriptase activity Like their intact counterpsarts, the -Cso of these
fragments are in the sub-nano molar ranges, 0.3-0.4 nM. In the dose level of the assays, they
exhibit little cytotoxicity In addition, the fragments also remain fully ative i their activities
on HIV integrase inhibition, HIV DLTR topological inactivation and ribosome inactivation.
Conclusion: These results suggest a possible explanation for the b.iislifiliy of these antiHIV agents following oral administration.
Dr. Sylvia Le e-Huang, Biochemistry Department, NYU Medical School, 50 I st Ave. New
York, New York 10016, USA
Mo.A.I 100
ANTI-HIV EFFECT OF PREGNANCY HORMONE, HUMAN CHORIONIC
GONADOTROPIN (hCG) AND j3hCG IN VITRO
Bourinbaiar A.S., Fruhstorfer, Eric C. Metatron, Inc., 31 Stuyvcsant St eet, New York, NY
10003, USA
Objective: To identify further the anti HIV mechanism of human chonoric i)ondotropin
(hCG) in vitro with the goal of using this pregnancy hormone in AIDS prevention and therapy
Methods: The scrial ten-fold dilutions of a and 3 subunits (range 100 p i t100 pg per ml)
and the intact hCG as a control (1000 - 0.01 IU/ml) were tested for the suppression of
viral synthesis in chronically infected ACH-2 lymphocytes and U I nonocytes. Following 3
days of incubation with hCG preparations, the supernatants of ACI-t1 aI iI tl cultures were
harvested and tested for p24 release by p24 ELISA and for suppression o f reverse transcriptase activity hCG preparations were also tested against cell-toe ceil transmission of HIV
resulting from a physical contact of virus infected lymphocytes with trop1roblasts - he fetal
cells lining the outer layer of placenta.
Results: 3hCG exerts more potent effect on HIV replication than a sut;unit. Select peptide
fragments of j3hCG molecule were also antivirally active. Although significant disparity has
been observed between U I and ACH 2 cells, in both cases the doe respoise curve
appeared to be U-shaped.This peculiar reaction is due to the enhlr erment of viral synthesis by extremely high doses of hCG- concentrations that are severs i orders cif magnitude
above the normal levels found in pregnant women.
Conclusions: hCG as well as j3hCG subunit were found to suppr'es HIV replication as well
as to prevent the spread of the virus in utero model.These findings suggest thsat hCG may
be responsible for the low rate of HIV transmission from mother to -fetus. Based on preliminary clinical trial it is likely that low doses of hCG could be usefil iri AIDSL prevention and
therapyThe therapeutic use of free 3hCG subunit is more promiin- since i has no effect
on reproductive function and is, thus, considered as biologically inert
Aldar S. Bourinbaiar; Metatron, Inc., 3 I Stuyvesant St., NewYork, 1 ii.1[ tUSA/ lel: 2 I 2 -260-8814 Fax: [phone] email:emballon@cpipoline,com
Mo.A.I 101
LITHIUM y-LINOLENIC ACID (LiGLA) - ASSOCIATED CYTOTOXICITY IN ACUTE
AND CHRONIC HIV INFECTION MODELS IN VITRO.
Mpanju, Onesmo, Manning, J., Randall*, S., Winther**, M., Montaner J. ( 'Shausghnessy M.,
Conway B. British Columbia Centre for Excellence in HIVIAIDS.VaV:ncov British
Columbia, Canada *St. Bartholomew's Hospital, London, U.K. and S ' colit Pharmaceuticals
(Canada) Ltd, Kentville, Nova Scotia, Canada.
Objective: To carry out a comparatsve study of LiGLA - associated t tPxi ty in chronic
and acute models of HIV infection.
Methods: (i) PHA stimulated PBMCs were infected with HTLV-IIIB or I 0 6 isolates of
HIV (MOI0.t00 0.05). Cells were then cultured in the presence of t),0 smL LiGLA.
Viable cells were enumerated by Trypan Blue dye exclusion every da s o I.I days, with
supernatants collected for p24 antigen determination at the sance time Ali naitively infected and uninfected cells were grown in parallel for 7 days without dru slIer vhih they
were placed in flesh viral culture medium in the presence of dru.-I tree ad seven days
later, viable cells were enumerated and supernatants collected as above. (s) As a model of
chronic infection, 8E5 cells (and the uninfected parent A3.01 cell) wl as he OM-10.1
cells (before and after stimulation with [NF-ot to induce active v ial -,pli 5on) were con
sidered. Cell viability and viral growth (as measured by p24 antge cv' n ulture super
natants) were considered nve 7 dart, in th e presence or absen' ot d y t
Results: I C]LA -associated differential cytotoxcsty was discernible rn cuts' y shorcted
PBMCs in which 10Jpg/mL LiGLA reduced viability by 12% and 3i% in te H hLV IIIB and
G910-6 - (MOI0.05) infected cells respectively measured 10 dae po-t inetion.Varying
the MOI did not significantly influence this observation. However',hen the i5flection was
allowed to become established for 7 days before addition of dru te 0, cytotoxic con

centration of LiCGLA was reduced from 20 to 10pg/mL in the HTLV-IIIB infected cultures
but remained at 3tag/mL and higher in the mock infected cells. G9 10-6 - infected cultures
showed a similar ind In the chronic infection model, 10tg/mL LiGLA reduced viability of
8E5 or stimulated O)M- 10. I cells by approx. 60% with little effect on A3.0I or unstimulated
OM 10.1 cells.
Conclusion: Chronically infected cell lines are more susceptible than acutely infected prima
rycells to iGLA -associated cytotoxioty even when both are actively producing progeny
virus. [his may be. of particular importance in vivo, in the design of a novel antiretroviral
agent aimed at viral tissue reservoirs of such chronically infected cells. Work is underway in
our centr e to elucidate the mechanism of action of LiGLA, within the context of a phase I
clinical t-ii in Canada.
Mr. Onesrmo F"Ipanjiu, British Columbia Centre for Excellence in HIV/AIDS, 667-1081
Burird St.Vancouve BCV6Z IY6.Tel: [phone] Ext. 3179; Fax: [phone];
Email: ompa, u0hivnet.ubc.ca.
Mo.A.I 102
ANTIVIRAL PROFILE OF HBYO97,A NONNUCLEOSIDIC INHIBITOR OF HIV-I RT IN
A PHASE I STUDY
R0bsamer 'ain2i ann H.-I, Wainberg, M.A.2, Huguenel, E3. Shah, A3, Paessens. AI, Kleim,
J. Rosn e I I. Institute of Virology Bayer Pharma Research Center, 42096 Wuppertal,
Germany 2. JewiMI General Hospital, Montreal, Canada, 3. Bayer Corp.West Haven, USA, 4.
Hoechsi AG, Frankfurt, Germany
Objective: hLis study was designed to characterize viral load and viral characteristics during, 14 d 'y trealt ent of HIV- I -positive individuals with HBY 097.
Method Blood,ias drawn from the patients at day 0, 14 and 21 (i.e. 7 days after the last
day under treatment with HBY 097).Viral load determinations were done by PCR and
bDNA.Virs u,,as vcultured from PBMC on human umbilical cord cells and analysed for phenotype and genotype.
Results PRsults of l viral load determination varied considerably between the methods, but
were qulitatively comparable. All viruses before and after therapy were of the noncytopathoer/i NSI or c/d, von Bri esen et al (1990) J.Virol. 187,597) phenotype with moderate or Ilsv replii.ation capacity. Genotypic analysis of viral RNA from serum, of cultured
virusesia t pi oviral DNA showed no induction of a relevant NNRTI resistance mutation,
however in orn patient, a NNRTI mutation was present before treatment. Results on pharr akokinec propersties of HBY 097 are given in an accompanying paper by Shah et al.
Acknowledgements: We would like to acknowledge the help by Dr. Immelmann, Analysis
Fra kfurt, crnsiy and Dr: Dietrich, Georg-Speyer Haus, Frankfurt, Germany in the deteranratin ') ' vi; al ()ad and resistance mutations.
I- Rubssarnen a igmann, PH-R Al II Virology BAYER AG, D-42096 Wuppertal
Teleph, ne: 0019 202-364143, Fax: [phone]
Mo.A.I 103
SYNERGISTIC INHIBITION OF HUMAN IMMUNODEFICIENCY TYPE I REPLICATION IN VITRO BY TWO AND THREE-DRUG COMBINATION OF DELAVIRDINE,
LAMIVUDINE AND ZIDOVUDINE
Keng 1:hcrg, Prul J. Pagano. Cancer & Infectious Diseases Research, Pharmacia & Upjohn,
Kalamazoo, Michigan 49001-0199 USA
Objective: To e',,aluate combination therapy involving Delavirdine (DLV), a non-nucleoside
reverse trans ciplase inhibitor, inr two- and three drug combination with Lamivudine (3TC)
and zidovsdine (ZDV).
Methods: Viral growth was assayed in peripheral blood mononuclear cells infected with a
mcolecul ily cloned clinical isolate (HIVJR-CSF). Drug synergy was estimated by the combination index method.
Results: Over a range of drug concentrations, and several ratios of drug combination, DLV
in twodrug combination with 3TC or ZDV synergistically (combination index <.0) inhibit
ed the replicatio of HIV IjR-CSF.A synergistic drug effect was also observed when all
three compounds were employed together. In general, there was greater suppression of
viral growth as the number of drugs in culture medium was increased from one to three.
None of these combination showed any significant additive or synergistic drug toxicity.
Conclusions: These in vitro synergy data support the potential use of DLV in two- or threedrug cohbr ation with 3TC and ZDV in patients with AIDS.
KT Chong, Cancer & Infectious Diseases Research, Pharmacia and Upohn, Inc., 30 I
Henretta St., Kalmazoo, MI 49001 USA Telephone: [phone] Fax: [phone]
Mo.A. I 104
ANTI-HIV ACTIVITY BY MGN-3 IN VITRO
Ghorneum Manmdooh. Drew University of medicine and science. Los Angeles, California
Objective: to examine the effect of MGN-3 on HIV induced syncytia formation (SF) In Vitro
M a-3 is an arabinoxylane from rice bran, that is enzymatically treated with an extract
from basidiomnycetes mycelia.
Method: Peripheral Blood Lymphocytes (PBL) from AIDS patients were cultured with PHA
is the pieaence or absence of different concentrations of MGN-3 (I12.5 - 100 ug / ml) for
7days. Te number of syncytia were evaluated, and the size of each was also recorded.The
effec of MtGN t on PLIA induced PBL proliferation was studied by MTT assay
Results: resatrent with MGN 3 resulted in: I) Signiicant inhibition in the SF, 2) The effect
was dose dependent, percentage of inhibition in SF was 38.5, 50, 62.5, and 75% at concentrations o c 2., 25, 50, and 100 ug /mli respectively; 3) Complete absence of SF having large
and median s ze post treatment, and 4) MGN 3 caused 25 30% inhibition of PBL proliferConclusion: W conclude that MGN-3 is a natural product that possesses a potent effect
aain 5t sy"u aormation by HIVThis propery of MGN 3 may be of potential value in
therapy of It1V, infection. Supported by Daiwa Pharmaceutical Co. Ltd.
[-1Mi. e as., 51 E. 120th Street, L.A., CA. 90059 Telephone: [phone]
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