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

368 Abstracts ThPeA7137-ThPeA7140

XIV International AIDS Conference

tion. To further understand the determinants and parameters of heterosexually
acquired HIV-1 disease, we investigated cellular activation and chemokine receptor expression on T lymphocytes,
Methods: Cervical Intraepithelial and peripheral blood mononuclear cells
(PBMC) were harvested in parallel from HIV+ women at different stages of
disease. Using flow cytometry T- lymphocyte subsets (CD4+ and CD8+) were
quantitated. Expression of activation markers (CD69, CD25, HLA-DR) and the
chemokine receptor CCR5 on T-lymphocytes were determined in matched cervical and blood preparations.
Results: The number of CD4 + T lymphocytes was decreased in both cervical
(p<0.05) and blood (P<0.05) samples with progressive HIV-1 disease coincidental with decline in CD8 T cells. Coincidentally, CD4+ and CD8+ T-lymphocyte activation markers were elevated (CD69 p<0.001, CD25 p<0.02, HLA-DR p<0.05).
In contrast, T lymphocyte chemokine receptor expression did not change in the
two compartments.
Conclusion: These findings suggest cervical mucosa supports productive HIV infection with HIV-induced cytopathicity leading to local CD4 lymphocyte depletion
and reduced capacity to respond to opportunistic infection. Increased exposure
to such infection drives T-lymphocyte activation allowing latently infected resting
cells to produce more HIV, thus, perpetuating the cycle.
Presenting author: Moses Kapembwa, Dept Immunology, Chelsea & Westminster Hospital, 368 Fulham Road, London SW10 9NH, United Kingdom, Tel.: +44
(0)20 8746 5987, Fax: [phone], E-mail: [email protected]
ThPeA7137 Evaluation of viral burden and immune cell
sub-sets in cervical epithelium of women receiving
combination anti-retroviral therapy (CART)
M. Prakash1, S. Patterson1, F Gotch', M. Kapembwa2. 1Imperial College,
London, United Kingdom; 2 Dept of GUM/HIV Medicine, Imperial College,
Northwick Park Hospital, Harrow, United Kingdom
Background: HIV disease progression is characterised by depletion of the CD4
lymphocyte subpopulation and increased plasma viral load(VL). Highly active
anti-retroviral therapy (HAART) effectively suppresses VL with resultant increase
in CD4 T cells. Whether similar change in VL and CD4 T-lymphocyte repopulation occur in cervical mucosa is unknown. We hypothesised that loss of cervical
intraepithelial (CIE) CD4 T cells and macrophages are features of HIV infection
and reflect continued viral replication at this site.
Methods: Endocervical samples were obtained using cytobrush from 28 HIVinfected, pre-menopausal women during midcycle. 17 received HAART and 11
were therapy-naive. 12 sero-negative women were controls. CIE leucocytes
were surface-stained using monoclonal antibodies and mononuclear cells bearing CD3, CD4, CD8 T cells; CD14 macrophage/monocyte markers determined
by fluorescent antibody cell-sorter analysis. For proviral DNA, cervical T cells and
macrophages were isolated by positive selection using Dynal beads and cells
lysed. DNA was purified and the proportion of CIE lymphocytes and macrophages
containing proviral sequences determined by limiting dilution nested PCR using
primers corresponding to the HIV-1 pol gene.
Results: CIE CD4 lymphocytes were decreased in women with VL >200 RNA
copies/ml compared with healthy controls patients (31+13 vs 91+22, p<0.05) resulting in reduced CD4/CD8 ratio (0.8+0.3 vs 2.2+0. p<0.02). Macrophage subpopulations were affected to a lesser extent. CIE Proviral DNA was detectable in
both T-lymphocytes and macrophages in almost all treated (13/15) women with
highest levels in those with VL >200 (180+74, 145+56 copies, respectively) compared to VL <200 (7+3,17+19 copies, p<0.01).
Conclusion: Cervical mucosa is a potential reservoir of HIV infection despite
'successful' HAART and drives local immunodeficiency including emergence of
drug resistant virus. New strategies are urgently required to eliminate HIV reservoirs.
Presenting author: Moses Kapembwa, Dept of GUM/HIV Medicine, Imperial
College, Northwick Park Hospital, Watford Road, Harrow, United Kingdom, Tel.:
[phone], Fax: [phone], E-mail: [email protected]
ThPeA71 38 Administration of HAART in primary and acute
HIV-1 infection induces transient specific T cell
responses
A. Pires', M. Nelson2, A. Pozniak2, B. Gazzard2, F Gotch', N. Imami'.
'Imperial College of Science Technology and Medicine, Dept Immunology
Chelsea & Westminster Hospital, 368 Fulham Road, London, United Kingdom;
2HIV & GU Medicine, Chelsea & Westminter Hospital, London, United Kingdom
Administration of HAART during acute/primary HIV-1 infection leads to augmentation of strong virus-specific helper T lymphocyte (HTL) responses. It is suggested
that this might be because potent anti-retroviral therapy protects the activated
virus specific HTL so that these cells are not lost during acute primary HIV-1 infection. Recently it has been shown that despite loss of functionality, detectable

interferon-y (IFN-g) producing HIV-1 specific cells persist in the peripheral pool.
Seven patients presenting with symptomatic acute HIV-1 infection were assessed
for: lymphoproliferation using 3H-TdR incorporation, antigen induced intracellular
IFN-g production by flow cytometry and detailed phenotypic analysis at baseline
and weeks 4, 12, 24, and 36 after initiation of HAART. Correlation with CD4+ T
cell counts and viral load was also carried out.
Lack of proliferation was seen in 6 of 7 patients early in HIV-1 infection despite

detection of HIV-1-induced intracellular IFN-g in 4 of 7 patients. Both proliferation
and IFN-g production were transient, with stronger responses at week 24 in 4 of 7
patients which inversely correlates with HLA-DR surface expression (r=-0.95). No
patient maintained the same response throughout the study. Phenotypic analysis
reveals hyperactivation at baseline as seen by high levels CD38 on both CD4+
and CD8+ T cells, which decreased on both T lymphocyte subsets during HAART
(p<0.01).
The CD4+ HTL dysfunction seen in HIV-1 infection early during infection is
marked by a loss of HIV-1-specific responses in terms of proliferation, which
appears distinct from the antigen induced intracellular IFN-g production. Upregulation of HLA-DR during immune hyperactivation is suggestive of T:T presentation which is associated with anergy. Early treatment with HAART reverses
the anergic state, however the transient pattern of the virus-specific responses,
and their long-term effect on the control of HIV-1 disease, remain to be elucidated.
Presenting author: Antonio Pires, Dept Immunology, Chelsea & Westminster
Hospital, 368 Fulham Road, London, United Kingdom, Tel.: [phone], Fax:
++442087465997, E-mail: [email protected]
ThPeA7139 Rantes pre-and post-3 y of HAART and Ccr5
expression in HIV-1 infected children
C.I. Perez-Maldonado', M. Hernandez2, I. Caragol2, J.M. Bertran3, T. Espanol2.
' Laboratorio de Inmunologia, Facultad de Farmacia, Universidad de Los Andes,
Correos de Merida, Venezuela; 2U. Inmunologia, Hospital Vail d'hebron,
Barcelona; 3U. Inmunodeficiencies, Hospital Vall d'Hebron, Barcelona, Spain
Background: Chemokines are proinflamatory cytokines that attract and activate
specific leukocyte subsets whose receptors are used as coreceptors by some
viral strains. Increased levels of RANTES have been associated with better prognosis and immune recovery.
Objectives: to evaluate the role of RANTES and CCR5 in immunological reconstitution of HIV-1 infected children under HAART.
Patients and methods: Lymphocytes subsets, CCR5 expression on CD4+ cells
and intracellular production of IL-2 and IFN y were determined by flow cytometry (FCM). RANTES levels were assesed by ELISA in plasma and lymphocyte
culture supernatants. 32 infected children (4- 12 y of age) and an age-matched
control group were studied.
Results: RANTES plasma levels were 886 and 1025 pg/ml pre and post HAART,
respectively Mean CCR5 expression on CD4+ cells was 18 % and lower in 3
cases heterozygous for CCR5 8 32 mutation. In the control group mean CCR5
expression on CD4+ cells was 38 % and plasma RANTES levels 687 pg/ml.
RANTES levels and CCR5 expression were also analyzed in relation to age and
no significant differences were observed. Intracellular IL-2 and IFN y values were
no significantly different from controls, although CD4+/CD8+ ratio was much lower
in infected patients even after therapy.
Conclusions: Lower CCR5 expression than controls and increased RANTES
levels post therapy correlate with the improvement in total CD4+ cells, clinical
conditions and decreased viral load in the post HAART period, and could be a
reliable marker of immune recovery.
Presenting author: Cesar Perez Maldonado, Ap. Postal 611, Correos de Merida,
Venezuela, Tel.: [phone], Fax: [phone], E-mail: cesarp01
@cantv.net
ThPeA7140 HAART allow the reconstitution of cytomegalovirus
(CMV)-specific cell-mediated immunity in
HIV-infected patients
M. Alfonzo Diaz', D. Blanc2, C. Troadec2, M. Eliaszewicz2, G. Gonzalez2,
D. Scott Algara'1. ' Institut Pasteur, 25 Rue du Dr Roux, Paris 75015, France;
2CIRBS, Hopital St. Joseph, Paris, France
Background: Because CMV retinitis could be observed after initiation HAART,
we aimed better understand how HAART reconstitute the CMV-specific immunity.
We prospectively studied patients receiving HAART whose CMV retinitis was quiescent in order to establish the degree of T lymphocyte reconstitution obtained
during one year of therapy.
Methods: We measured the number of circulating T-cells and their subsets,
the plasmatic HIV-RNA concentration, the specific proliferation cellular (LPR),
the specific secretion of cytokines (IL-2, IL-4, IL-5, IL-10, IFN-gamma and TNFalpha), and the cytolytic activity of CD8+ T cells against autologous fibroblasts
infected with CMV. All tests were done prospectively at five time points during
follow-up: baseline, 3 months, 6 months, 9 months and 12 months.
Results: In untreated patients, we found a CMV-specific CTL activity despite
the absence of CMV-specific LPR, secretion of IFN-gamma, TNF-alpha and IL2. However, the reconstitution of these CMV-specific responses was observed
(LPR and cytokines secretion) after 6 months of HAART simultaneously with a
significant increase of CMV-specific CTL activity and a suppression of HIV load.
Moreover, the reconstitution of CD4+ T cells during HAART, showed a character
istic delay of increase in naive CD4+ T cells compared to the memory CD4+ T
cells.
Conclusions: Our findings suggests that one year of HAART, allowed the recovery of various CMV-specific responses, probably by a redistribution of memory
CD4+ T cells in our cohort of patients. Measurements of CMV-specific responses
during the reconstitution by HAART are needed to reduce the risk of CMV disease
when the anti-CMV therapy could be discontinued in HIV-infected patients.