Bridging the Gap: Conference Record [Abstract book, International Conference on AIDS (12th: 1998: Geneva, Switzerland)]

12th World AIDS Conference

Abstracts 22282-22286 319

620*/22282 HHV-8, proliferation, apoptosis and ploidy in
epidemic (AKS), endemic (EKS), and classical
(CKS) Kaposi's sarcoma (KS)
Peter Biberfeld1, Epahata Kaaya2, E. Castanous-Velez1, T. Heden1, M. Stuzl3,
J.Z. Zou4, J.N. Kitinya2. 1Karolinska Institute and Hospital, S 171 76 Stockholm;
3MTC Karolinska Institute, Stockholm, Sweden; 2Muhimbili Medical Hospital,
Dar Es Salaam, Tanzania;, Max-Planc Institute, Martinsried, Germany
Objectives: To study the possible interrelationship of HHV8 infection, cell proliferation, apoptosis and ploidy in AIDS associated (AKS), endemic Kaposi's sarcoma
(EKS) and classical Kaposi's sarcoma (CKS).
Design: Prospective, controlled study.
Methods: Formalin fixed and frozen biopsies of KS were studied by PCR
and in situ hybridization (ISH) for HHV8-DNA and sequencing of the KS330
Bam fragment. Ploidy was done by flow cytometry of cell nuclei from KS spindle
cells (SC). Immunohistochemistry was done for Ki67, bcl-2, c-myc, Mcl-1, CD40,
CD40L, CD95 and CD95L expression. Apoptotic cells were evaluated by the
TUNEL method.
Results: By PCR all KS biopsies contained HHV-8 DNA, which was shown by
ISH to be associated predominantly with the SC of KS. No consistent mutations
distinctive for African vs Swedish KS were detected. All KS types showed a diploid
or near diploid DNA content and S-G2 analysis showed a moderate proliferation
level (1.6-8.9%). All KS types expressed bcl-2 and c-myc but stronger in the SC
of nodular KS compared to patch or plaque stages. The Bax protein was weakly
but similarly expressed in SC as compared to bcl-2. The TNF/NGF receptor family
and ligands showed strong SC reactivity for CD40, weak to moderate for CD95
but not CD40 or CD95 ligands. KS lesions showed low rates of apoptotic cells
(- 1%) in most cases, which appeared to be associated with lymphoid cells.
Conclusion: The association of HHV8 with all types and stages of KS suggests
an important, but unclear pathogenic role for this novel virus. The KS lesions
appear to evolve through moderate proliferation and low apoptotic activity of SC
and other cells with diploid DNA content, suggesting a multifactorial reactive,
rather than clonal pathogenic mechanisms in KS evolution.
F619*/22283 North American phase 3 study (Protocol
L1057T-31) of Panretin.. gel (LGD1057, ALRT1057)
for cutaneous AIDS-related Kaposi's sarcoma
Alvin Frie Dman-Kien- 2, M. Conant3. New York University, Dept. of
Dermatology 550 First Street, New York, NY; 2New York University Medical
Center New York NY; 3Private Medical Practice San Francico CA, USA
Background: A Phase 3, multicenter, randomized, double blind clinical trial was
conducted to evaluate the safety, cutaneous tolerance and anti-tumor effects
of PANRETIN (9-cis-retinoic acid) Gel in patients with AIDS-related Kaposi's
sarcoma (KS).
Methods: HIV-positive patients with biopsy proven KS were randomized to
topical PANRETIN 0.1% Gel or vehicle gel, with intended treatment at least 12
weeks, starting TID advancing to QID. Six index cutaneous lesions and other
parameters were assessed at least every four weeks.
Results: 35 US and 3 Canadian study centers enrolled 266 male and 2 female
patients. Treatment arms (PANRETIN Gel vs. vehicle gel) were well matched
for age, ethnicity, gender, time since KS diagnosis, Karnofsky status, history of
opportunistic infections, KS lesions, presence of visceral KS, baseline CD4 count
(median 154 vs. 144 cells/mm3), % receiving prior systemic (36% vs. 34%) and
prior topical/local (49% vs. 53%) anti-KS therapy, and % receiving concurrent
antiretroviral (ARV): any ARV (93% vs. 93%), 3 or more ARV (72% vs. 79%),
and protease inhibitor (74% vs. 82%). The overall response (complete or partial)
rate determined by applying AIDS Clinical Trial Group (ACTG) criteria to each
patient's 6 index lesions was 35% (47/134) for PANRETIN Gel vs. 18% (24/134)
for vehicle patients (p = 0.002). The cumulative response rate for all patients
treated with PANRETIN Gel including the follow-on, open label phase was 49%
(90/184). One patient had complete clearing of all index lesions on PANRETIN
Gel. While responses for the vehicle arm patients correlated with the degree of
ARV therapy, after adjusting for concurrent ARV therapy the response rate for
PANRETIN Gel was still substantially higher than for vehicle. The Physician's
Global Assessment (PGA) responses were substantially higher for PANRETIN
Gel vs. vehicle (p = 0.000003). Response according to ACTG criteria was strongly
corroborated not only by PGA, but also by the patient quality of life responses.
PANRETIN Gel-related adverse events were nearly exclusively confined to the
site of application, with the most frequent consisting of rash (72% incidence), pain
(32%) and pruritus (13%).
Conclusions: PANRETIN Gel produced responses according to ACTG criteria in nearly 50% of patients treated, and was statistically superior to vehicle
gel in the initial blinded phase of study, even after adjusting for ARV therapy.
Adverse events were nearly exclusively confined to the site of application and
were generally easily manageable. PANRETIN Gel may provide a very tolerable, non-invasive, patient-controlled alternative to systemic chemotherapy for the
treatment of cutaneous AIDS-related KS.

22284 Genetic abnormalities in Kaposi's sarcoma tumors
Charles Barkin1, S. Janz2, S. Pack3, A. Coleman3, E. Musaba4, R. Biggar1,
Z. Zhuang3. 1 Viral Epidemiology Branch, NCI EPN/434 6130 Executive Bldg,
Rockville, MD; 2Laboratory of Genetics, Bethesda, MD; 3Lab. of Pathology, NCI,
Bethesda, MD, USA; 4Univ. Teaching Hospital, Lusaka, Zambia
Background: Kaposi's sarcoma is a complex tumor with features of both hyperplastic proliferation and neoplastic growth. We have used a variety of molecular
techniques to investigate genetic abnormalities underlying this disorder.
Methods: To determine if Kaposi's sarcoma is monoclonal, we used the HUMARA assay to assess methylation patterns in tumor DNA from female patients
with AIDS-related disease. Clonal relatedness was determined by comparing
the inactivated alleles in multiple lesions from individual patients. Alterations in
genomic content was investigated by comparative genomic hybridization. Loss of
heterozygosity was analyzed with selected microsatellite markers. DNA for these
studies was obtained from spindle cells isolated by microdissection in order to
increase the sensitivity of these techniques.
Results: Microdissected tumor DNA showed highly skewed methylation (i.e.,
predominant methylation of one HUMARA allele) in most evaluable tumors. Multiple lesions from individual subjects shared the same methylated allele, and no
patient had tumors with discordantly skewed methylation. Comparative genomic
hybridization on directly examined samples has been unsuccessful because of
inadequate DNA recovery, but further studies are ongoing utilizing DNA amplified
by the polymerase chain reaction. No loss of heterozygosity was detected with a
limited number of markers.
Conclusions: Kaposi's sarcoma is a monoclonal neoplasm, at least in the setting of HIV infection. Multicentric tumors originate from a single clone of precursor
cells that circulate to multiple sites and proliferate. Further studies are needed
to identify the genetic changes permitting neoplastic transformation and clonal
outgrowth.
22285 Production of human /3-chemokines in AIDS-Kaposi's
sarcoma patients
Maria Caterina Sirianni1, E. Scala2, L. Vincenzi1, S. Topino1, M. Capuano3,
F. Aiuti1, B. Ensoli4. 1 Chair of Allergy and Clinical Immunology, Viale Dell'
Universita' Roma; 2lstituto Dermopatico Immacolata, Rome; 3Dermatology
Catholic University, Rome; 4lstituto Superiore Di Sanita; Virology, Rome, Italy
Background: Kaposi's sarcoma (KS) is the most commom neoplasm of AIDS
patients. At least two viruses, HIV-1 and the human herpesvirus 8 (HHV-8).
whose genome encodes for some cytokines and f chemokines, are involved in
its pathogenesis. The aim of the study was to assess the production of the human
Macrophage Inflammatory Protein la (MIPl) and t (MIP1t) as well of RANTES
[regulated-upon-activation, normal T expressed and secreted] in peripheral blood
CD8+ cells and in skin cultures from KS patients.
Methods: 53 clones were obtained from 3 AIDS-KS patients in the presence
of allogeneic feeder cells and of recombinant Interleukin-2 (rlL-2), after isolation
of peripheral blood mononuclear cells on a Ficoll Hypaque gradient and subsequent expansion in rlL-2. In parallel, cultures were set up from skin biopsies of
9 AIDS-KS and 9 Classical KS (C-KS) lesions and from 3 uninvolved skin of
C-KS patients. Skin cultures were established in the presence of rlL-2 enriched T
conditioned medium (TCM). Peripheral and skin lymphocytes were phenotyped
by monoclonal antibodies to CD3, CD4, CD8, CD19, CD16/56 surface antigens
and to the,u/ chains of the T Cell Receptor. Human chemokines as well as human ylnterferon (ylFN) and IL-4 cytokines were measured by ELISA (Quantikine,
R&D System, Minneapolis, MN).
Results: The majority of the clones from AIDS-KS patients presented a
u/Ip+CD3+CD8+ phenotype and were mainly secreting ylFN as well as MIP1
proteins. Rare production of RANTES was found. The same spectrum of cytokine
and chemokine production was found in TCM-expanded cultures of lesional skin
lymphocytes, whose phenotype was also,u/ý+CD3+CD8+. This was observed
both in AIDS-KS and C-KS.
Conclusions: HHV-8 encoded chemokines and cytokines are poorly expressed
in KS tissue, whereas human cytokines and chemokines appear to be highly produced. In macrophages from the same skin cultures and in the clones from one
AIDS-KS patient HHV-8 DNA sequences were found. Thus, it appears that type
1 human cytokines and chemokines are not effective in the control of HHV-8
replication, contributing to further recruitment and adhesion of inflammatory cells.
with disease amplification.
22286 Kaposi's sarcoma: A new approach to follow-up using
epiluminescent light microscopy
Marl Delhere1, Joachim Krischer1, T.L. Trellu Laurence2, Sonia Roten1,
Jean-Hilaine Saurat1, Bernard Hirschel2. 1Department of Dermatology, Geneva
Hospital 1211-Geneva 14; 2lnfectious Disease Unit/Geneva Hospital Geneva,
Switzerland
Background: AIDS Clinical Trials Group (ACTG) criteria are widely used to

follow-up HIV-related Kaposi's sarcoma (KS); they are based on macroscopic
observation. Epiluminescent light microscopy (ELM) is a non invasive method
designed to improve diagnostic accuracy of pigmented skin lesions. Vascular tumors also exhibited ELM features which can be precisely measured and followed.
The aim of this study was to assess the suitability of ELM in KS as a tool for
lesional follow-up during local and systemic treatments.