Abstract Book Vol. 2 [International Conference on AIDS (14th: 2002: Barcelona, Spain)]

26 Abstracts WePeA5798-WePeA5801

XIV International AIDS Conference

of ET-1-induced effects on BBB permeability was examined. Particular emphasis was given to the paracrine effects of ET-1 on IL-1 beta production by human
astrocytes.
Methods: A recently developped a human in vitro BBB model was used. Real
time RT-PCR and ELISA methods were used to demonstrate the synthesis and
production of ET-1 and ILlbeta. BBB permeability was assessed using a paracellular marker,sucrose. The expression of cyclooxygenase-2 (Cox-2) was determined by flow cytometry analysis.
Results: We showed that TNF-o induced productioon of ET-1 in an in vitro model
of human BBB. Both TNF alpha and ET-1 induce permeability of human brain EC.
This effect was associated with TNFalpa induced IL-1 beta syntheis by astrocytes.
treatment of human primary astrocytes in a single culture with ET-1 induced IL1beta syntheis associated with the Cox-2 expression in these cells.
Conclusions: We therefore suggest that the paracrine actions involving ET-1,
TNfalpha and IL-1beta between astrocyte cells and brain EC may play a relevant
role in BBB breakdown during CNS inflammation.
Presenting author: aloise mabondzo, drm/dsv/spi, cea saclay, 91191 gif sur
yvette cedex, France, Tel.: [phone], Fax: [phone], E-mail: aloise.
[email protected]
WePeA5798 Interleukin-6 co-operates with a new type I IFN,
IFN-tau, to inhibit early steps of HIV-I biological
cycle
C. Rogez', P. Clayette2, M. Martin3, N. Dereuddre-Bosquet2, J. Martal4,
D. Dormont3. 1CEA, INRA, svn/dsv/drm, cea, 60-68 avenue de la division leclerc
bp 6, 92265 fontenay-aux-roses cedex, France; 2SP-BIO, Fontenay-aux-Roses,
France; 3CEA, Fontenay-aux-Roses, France; 4INRA, Jouy-en-Josas, France
Background: Type I interferons (IFN) exhibit efficient antiviral activities notably
against HIV. IFN-alpha has been used as a treatment in AIDS-associated Kaposi's sarcoma, but its side effects restrict its clinical uses. IFN-tau is an ovine
or bovine non-cytotoxic type I IFN which displays higher inhibitory effects towards HIV replication than IFN-alpha, particularly in human monocyte-derived
macrophages (MDM). The antiretroviral activity of IFN-tau seems to involve antiviral and immunomodulatory mechanisms: IL-6 synthesis is increased in dosedependent manner in MDM treated with IFN-tau and a specific inhibition of IL-6
biological activity decreases the antiretroviral efficiency of IFN-tau.
Methods: Herein, the effects of IFN-tau on early steps of HIV biological cycle in
MDM were investigated by amplifying intracellular HIV RNA 1 hour after infection
and by quantifying intermediate reverse transcription (RT) products. Host cell antiviral factors, 2',5'-OAS, PKR and protein MxA were measured by RT-PCR and
Western Blot.
Results: After a one-hour infection, a significant decrease of intracellular HIV
RNA amount was found in MDM treated with IFN-tau. In parallel, no additive inhibition was observed with IFN-tau during the elongation of proviral DNA. These
results suggest either an inhibition of HIV nucleocapsid uptake or an immediate
HIV RNA degradation, and the expression of 2',5'-OAS, MxA protein and PKR
was then measured. IFN-tau induced the expression of these three host cell factors. The role of IL-6 on these different steps was evaluated and we showed that
IL-6 co-operates with IFN-tau during the very early steps of HIV biological cycle.
Indeed, the effects of IFN-tau on intracellular HIV RNA were decreased when IL-6
biological activity was neutralised.
Conclusion: Altogether, these results evidence that IFN-tau uses the same antiretroviral pathway as others type I IFN in MDM, and that IL-6 takes part to its
inhibition of early steps of HIV biological cycle.
Presenting author: christine rogez, svn/dsv/drm, cea, 60-68 avenue de la division leclerc bp 6, 92265 fontenay-aux-roses cedex, France, Tel.: [phone],
Fax: [phone], E-mail: [email protected]
WePeA5799 Low-dose rHuGM-CSF for the treatment of chronic
neutropenia in HIV-infected patients treated with
nucleoside analogues based HAART regimens and
concomitant prophylaxis for opportunistic
infections
A. Mastroianni, C. Cancellieri, F Allegrini, S. Pignatari. Division of Infectious
Diseases, "G.B. Morgagni" General Hospital, Piazza Solieri 1, Forli, Italy
Objective: The aim of this study was to investigate the use of long-term treatment
with low-dose rHuGM-CSF to prevent severe neutropenia and related infectious
complications in leukopenic patients (pts) treated with nucleoside analoguesbased HAART regimens and prophylaxis for opportunistic infections.
Methods: From January 1997 through December 2001 we enrolled in this study
28 HIV+ pts (26 males, 2 females, range of age 26-53 years) with a mean absolute neutrophil count (ANC) of 600/L (range, 500-800), recognized at high risk
to develop severe neutropenia and related infectious complications.Two delivery
schedules were tested.In the first type a total weekly dose of 600 mcg (300 mcg

every 3 days) of rHuGM-CSF was delivered in 17 pts with an ANC >500<700
neutrophils/mmc at baseline.We further tested a second dose schedule of 150
mcg of rHuGM-CSF administered two times weekly every three days in 11 pts
with an AN >700<900 neutrophils/mmc.ANC was eveluated weekly during the
first month and every two weeks subsequently. All pts were treated with nucleoside analogues-based HAART regimens associated with use of TMP-SMX,
and/or pyrimethamine,and/or ganciclovir, and/or amphotericin B.

Results: Pts received rHuGM-CSF for 4-32 weeks (median 8 weeks) without
loss of effect. Overal a complete response defined by a stable ANC of 1,500 neutrophils/mmc was observed in all cases. rHuGM-CSF-related side effects were
recognized in 65% of pts, including fever and bone pain. No pt presented severe neutropenia or infectious complications.No pt presented a modified HIV viral
load,while there was evidence of a moderate increase of the CD4+ lymphocytes
count.
Conclusion: The results of this study suggest that long-term treatment with
lo-dose rHuGM-CSF may reverse drugs-related neutropenia in pts treated with
HAART and prophylaxis for opportunistic infections. Long-term immunotherapy
with rHuGM-CSF may also induce an improvement in the CD4+ cells count in pts
receiving HAART regimens.
Presenting author: Antonio Mastroianni, G.B. Morgagni General Hospital, Piazza Solieri 1, Forli, Italy, Tel.: [phone], Fax: [phone], E-mail:
antoniomastroianni @yahoo.it
WePeA5800 Role of cytokines and C-reactive protein in the
diagnosis and outcome of HIV-1 infected patients
with pulmonary infections
N. Benito', A. Moreno', X. Filella2, J.M. Mir61, J. GonzIlez3, T. Pumarola3,
M.E. Vails3, M. Luna', A. Rah64, A. Torres4, J.M. Gatell'. 'Service of Infectious
Diseases. Institut Clinic Infeccions i Immunologia. Hospital Clinic, Hospital
Clinic, Villarroel 170, 08036 Barcelona, Spain; 2Centre de Diagnostic Biomedic.
Hospital Clinic, Barcelona, Spain; 3Service of Microbiology Institut Clinic
Infeccions i Immunologia. Hospital Clinic, Barcelona, Spain; I Service of
Pneumology Institut Clinic de Pneumologia i Cirugia Toracica. Hospital Clinic,
Barcelona, Spain
Objective: To evaluate the role of several cytokines and C-reactive protein (CRP)
in the diagnosis and outcome of HIV patients with pulmonary infections (PI).
Methods: Prospective study of all HIV patients with PI in our institution between
April '98 and May 2001. Plasma CRP, IL-10, IL-6, IL-8, IL-10 and TNF-o were
determined at admission and five days later. Patients were included in a protocol
addressed to study the etiology and outcome.
Results: 249 PI were diagnosed in 220 patients (160 males). The main etiologic groups were: bacterial pneumonia (BP) in 114 episodes, Pcarinii pneumonia (PCP) in 37 and tuberculosis (TB) in 36. Twenty-four (10%) patients died: 6
PCP cases, 4 BP and 1 TB. Median levels at admission of CRP and cytokines for
BP, PCP and TB were: CRP 10.2, 3.8 and 5 mg/dL respectively (p=0.0001); IL-8:
19, 3 and 2.9 respectively (p=0.045); TNFo: 46.5, 44 and 75 pg/mL respectively
(p=0.029). The sensitivity, specificity, negative predictive value (NPV) and positive
predictive value of CRP > or =10 plus IL-8 > or =20 for BP vs. PCP and TB were
69%, 83%, 71% and 82% respectively. The NPV of TNF > or =60 for TB vs. PCP
and BP was 93%. Concerning the levels of IL-11, IL-6 and IL-10, there were no
differences among the three etiologic groups. There were no correlation between
etiologies and cytokines and CRP values determined on the 5th day from admission. Compared with survivors, patients who died had at admission higher levels
of IL6: 95 vs 36 (p=0.014), IL-10: 22 vs 6 (p=0.01) and TNFo 70 vs 46 (p=0.041).
At day 5 post-admission, the IL-6 remained statistically higher in those patients
who died (126.5 vs 24; p=0.043).
Conclusions: HIV patients with BP had higher plasma CRP and IL-8 levels at
admission than PCP and TB patients. TNFo was higher in those with TB. Patients with PI and a worse outcome had higher levels of IL-6, IL-10 or TNF-o at
admission and maintained high IL-6 levels on the 5th day
Presenting author: Natividad Benito, Hospital Clinic, Villarroel 170, 08036
Barcelona, Spain, Tel.: [phone], Fax: [phone], E-mail: nbenito
@clinic.ub.es
WePeA5801I Production of macrophage inflammatory proteins
MIP-Ice, MIP-113 and RANTES in HIV-1 infection
from antigen-stimulated seminal cells and whole
blood: correlation with Thi type cytokine
responses
C. Panaiotidi', M. Kotsianopoulou', D. Papadopoulou', G. Panos2,
A. Roumeliotou'. 'National School of Public Health, AIDS Reference Center,
Athens, National School of Public Health, 196 Alexandras Avenue, 115 21,
Athens, Greece; 2,H/V UNIT Second Dept. of Internal Medicine, 1st /KA
Hospital, Penteli, Athens, Athens, Greece
Introduction: The effect of 1-chemokines in the seminal plasma on the transmission of HIV-1 remains uncertain. Higher RANTES levels in semen may increase
HIV concentration in the seminal inoculum by the recruitment of target cells or
may reduce the ability of M-tropic virus to infect recipient host cells.
Objective: To evaluate the potential clinical relevance of the 1-chemokines in
HIV-1 infected, and correlate the results with the Interleukin level of the same
samples.

Method: Peripheral blood and semen were collected from 40 HIV seropositive
patients, selected cross-sectionally, and 10 negative controls and stimulated in
vitro with (PHA). We measured the levels of RANTES, MIP-lca, MIP-1t, IFN-y,
IL-4 and IL-10 from the supernatants of the cell cultures using ELISA assays.
Results: We found that changes in the cytokine profile in blood and semen reflected different stages of the HIV-1 disease and correlated to the production of
l-chemokines. Seminal cell supernatants stimulated with PHA, and gave produc