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

12th World AIDS Conference

Abstracts 31178-31182 531

31178 Effects of triple antiretroviral therapy on the expression
of Fas antigen in patients with HIV1 infection
Rui Victorino1, Ana Sousa2, A.F. Chaves2, M. Doroana3, F. Antunes3,
R.M.M. Victorino4. 1Medicina 2 Piso 2 Faculdade De Medicina AV Prof Egas
Moniz 1600 Lisboa; 2Cel Immunology Lab Faculty of Medicine Lisbon; 3Dep
Infectious Diseases FML-HSM Lisbon; 4Medicine 2-Cel Immunology Lab
FML-HSM Lisbon, Portugal
Background: Massive activation of the immune system is considered to play
a role in the pathogenesis of HIV immunodeficiency. The expression of the cell
surface protein Fas (CD95) is enhanced upon T cell activation and is associated
to the initiation of apoptosis after binding to the Fas ligand. We investigated here
the modulation of Fas by highly active antiretroviral therapy (HAART).
Methods: Fourteen HIV1 infected patients with mean CD4 count of 378//1l
(range 11 to 828) and a mean viral load of 75.174 RNA copies/ml (range
16.700-588.820) were treated with lamivudine, stavudine and nelfinavir. Expression of CD95 within different T cell subsets was assessed by flow cytometric
analysis performed on peripheral blood mononuclear cells at day zero and 2, 4,
6, 8, 16, 24 and 32 weeks of therapy. These results were correlated with the CD4
and CD8 counts, viral load and lymphocyte proliferative responses to PMA+I,
PHA, PWM, anti-CD3, anti-CD3+anti-CD28 and tetanus toxoid assessed by 3H
thymidine incorporation.
Results: All patients responded to therapy both in terms of viral load and CD4
count. At time zero there was an increased frequency of CD95+. A very significant
and progressive effect of HAART on the expression of Fas was observed which
was most marked for CD4 T cells: 78.1% (time zero), 72.4%(week 2), 67.7%(week
4), 59.5%(week 8), 55.1%(week 16), 53.5%(week 24). The magnitude of the decrease was independent of the initial CD4 counts as well as of the viral load, and
varied in the 14 subjects between 4.8% and 46.6% at week 24. In CD8 T cells the
reduction was from 85.5% to 70.4% and in total CD3 cells from 83.2% to 64.6%
at week 24. These reductions were concomitant with a progressive recovery of
lymphocyte proliferative responses.
Conclusion: The use of HAART in patients with viral loads above 15.000 and
no evidence of opportunistic infectious is associated to a marked reduction in the
T lymphocyte Fas expression even in patients with well preserved initial CD4 cell
count, which may contribute to the improvement of T cell function and possibly to
a reduction in activation induced cell death.
S31179 Immunological profile of HIV infected patients at the time
therapeutic follow up is envisaged in Yaounde
(Cameroon)
Clement Bertin Ndongmo, L. Zekeng, Y. Deloko, S. Kuate, A. Tamokoue,
P. Kenmogne, L. Kaptue. Hematology-Immunology Laboratory, Chu-Yaounde,
Cameroon
Introduction: Continuous assessment of the immune status, especially CD4
T-cells counts is essential in monitoring HIV disease progression. Measurement
of CD4 T-cells levels has also proved essential for clinical staging, for epidemiologic studies and for decision regarding prophylactic therapies for opportunistic
infections and the commencement of antiretroviral therapy.
Objective: To assess the immune status of HIV infected patients at the time
therapeutic follow up is envisaged.
Methods: Included in the study were known HIV patients referred to our
laboratory for CD4 counts by clinicians in view of therapeutic follow up. After
recording socio-demographic and clinical data of each patient, about 10 mls of
blood were drawn into EDTA tube. Samples were collected between 8 a.m and
1 p.m and analyzed within 6 hrs of collection for CD4, CD8 and CD3 T-lymphocytes by flow cytometry using FACScount (Becton Dickinson) immunocytometer.
Statistical analyses were done using Microsoft Excel 5.1 and Epi Info 5.
Results: Three hundred and forty seven samples were analyzed between April
96 and December 97. The patients were made up of 193 (55.67%) male and 154
(44.33%) female with a mean age of 39.8 and 32.7 respectively. According to CDC
classification, 178 (51.3%) of the patients were at an advanced stage with CD4
counts <200 cells/mm3, mean value 70.9 cells/mm3; 119 (34.3%) intermediate
cases, CD4 counts between 200 and 500, mean value 334.8 and only 50 (14.4%)
early cases with counts >500, mean value of 647.7. The rate of CD4 count
depletion was significantly greater in male than in female, p = 0.002 (<0.05).
Conclusion: These figures show that therapeutic follow up of HIV infected patients is generally envisaged at an advanced state of immunodeficiency because
most patients lack access or present to the hospital at a later stage of the disease.
Some, even when diagnosed earlier cannot afford the cost of treatments or delay
seeking care and only come back at the last stage. While viral load quantitation
techniques are not yet available, CD4 T-cells measurement remains the only tool
used for therapeutic management of HIV infected patients in Cameroon.
31180 CMV disease status and CMV-specific lymphocyte
responses measured by flow cytometry in HIV-1-infected
subjects

Joseph M. Mc Cune, Krishna V. Komanduri, M.N. Viswanathan, E.D. Wieder,
D.K. Schmidt, B.M. Bredt, M.A. Jacobson. Gladstone Institute of Virology &
Immunology PDO. Box 419100, San Francisco, CA, USA
Objectives: To correlate CMV-specific CD4+ lymphocyte responses measured
by a flow cytometry-based assay with CMV end organ disease (EOD) status in a
cohort of HIV-1-infected subjects.

Design: Cross-sectional clinical study.
Methods: We have optimized a flow cytometry-based method of quantitating
CNW-specific CD4+ lymphocyte responses. Peripheral blood mononuclear cells
(PBMC) are isolated from whole blood and stimulated ex vivo with CMV antigens in
the presence of antibody to CD28 (for costimulation) and brefeldin A (to inhibit cytokine secretion). Cells are then fixed, permeabilized and stained with monoclonal
antibodies specific for CD4, CD69, and for intracellular cytokines (interferon-y
and TNF-u). Responses are analyzed by multiparameter flow cytometry by determining the frequency of CD4+CD69+ cells with detectable intracellular cytokine
production.
Results: 21 HIV-1-infected subjects were assessed, including 13 CMV-seropositive subjects with no history of EOD, 3 with active EOD (2 with retinitis, one with
confirmed CMV encephalopathy) and 5 with quiescent CNW disease after initiation of highly active antiretroviral therapy (HAART). All subjects without a history of
EOD demonstrated strong responses to CMV (range 0.70-49%). In contrast, none
of the subjects with active EOD had significant CMV-specific responses (range
0-0.16%, p = 0.009 relative to patients with no history of EOD). Responses of the
patients with quiescent retinitis after HAART were significantly higher than those
with active EOD (range 0.2-3.01%, p = 0.025) and not significantly lower than
those without a history of EOD (p > 0.05).
Conclusion: Loss of CMV-specific CD4' lymphocyte responses strongly correlates with the presence of active CMV end organ disease in HIV-1-infected
subjects. In contrast, patients with no history of EOD and those with quiescent
EOD after HAART demonstrated CMV-specific CD4~ lymphocyte responses.
These data suggest a role for this assay in clinical risk assessment and in studies
of immune reconstitution.
S31181 Coexpression of CD69 and CD28 antigens in
T lymphocytes of HIV-1(+) individuals
Helen Choremi-Papadopouldu1, Kostas C. Pitsios1, Vassilis Viglis1,
Konstantina Zissi1, Elisa Samouilidi1, Nikos Panagiotou1, Theodore Kordossis2.
1Immunology Dept. Laiko General Hospital, Agiou Thoma 17; 2Dept. Pathologic
Physiology Medical School, Athens, Greece
Introduction: CD69 antigen is one of the earliest markers expressed on activated
T lymphocytes following stimulation by a variety of mitogenic antigens. CD28 is a
costimulatory molecule necessary for the proliferation of T lymphocytes through
CD3/TCR. After stimulation of T lymphocytes expression of CD69 is increased
but this increase is lower in HIV-1(+) individuals compared to normal controls. As
we and others have shown CD28 is also decreased in T lymphocytes with the
progression of HIV-1 disease. In this work we studied the expression of CD69 in
CD28+ T lymphocytes after stimulation in HIV-1 infection.
Methods: We used the whole blood culture method stimulated with anti-CD3
monoclonal antibody and phytohemagglutinin (PHA). Triple immunofluorescence
was used for the coexpression of CD3CD69CD28 antigens (%) by FACSCalibur.
We studied 25 HIV-1(+) individuals of different stages (CDC 1993) and 10 normal
controls.
Results: HIV-1(+) individuals have much lower coexpression of %
CD69++CD28+ antigens after stimulation of lymphocytes with both anti-CD3
= 16.2 ~ 8.5 and PHA = 9.5 ~ 6.5 compared to normal controls anti-CD3 = 34
~ 12.5 (p < 0.0001) and PHA = 37.7 ~ 21 (p < 0.0001). The % CD69 in CD28+
T lymphocytes was in HIV-1(+) individuals after stimulation with anti-CD3 = 40 ~
18.6 and with PHA = 28.3 ~ 16.6 compared to normal controls 70 ~ 17.9 (p <
0.0001) and 77.4 ~ 23.3 (p < 0.0001) correspondintly.
Conclusion: After stimulation the coexpression of CD69 and CD28 was significantly lower in HIV-1(+) individuals compared to controls. These results suggest
that the coexpression of these antigens may be used as a marker to monitor
HIV-1(+) patients.
31182 Tolerance of skin allografts in advanced HIV infected
patients
Raymond Beaulieu1, B. Lamtu2, M. Cadotte2. 13840 St-Urbain Montreal Quebec
H2W 1T8; 2CHUM-Campus Hotel-Dieu, Canada
Objectives: Assessement of T-cells in HIV infected patients has been done in
several ways but there is no single screening test which if positive, categorically
rules out a significant defect or, if negative, is diagnostic forT-cell dysfunction
although skin grafts come close to fullfilling these criteria.
Methods: Dermatologic skin biopsy punches were employed. A 4 mm fullthickness skin punch was removed from the volar aspect of the patient's forearm.
A 5 mm split-thickness skin punch was similarly removed from a healthy HLA
mismatched but ABO compatible HIV negative donor and placed in the prepared
recipient site. Pressure was applied and care taken to eliminate hematoma formation. The graft was covered with a pressure dressing for 5 days. Thereafter, the
graft was covered with light gauze and observed three times a week for 3 weeks
and subsequently every month for 3 months and thereafter every 2 months, for
signs of rejection.
Results: Six HIV infected patients with less than 100 CD4 cell count were

studied (4 males and 2 females). Macroscopic and microscopic examinations
were done and no sign of rejection was observed after 4 months for one patient since he died of PCP. Rejection was not seen in an other patient up to
his death 8 months later due to a primary brain lymphoma. A third one died
18 months later of wasting syndrome and the skin graft was still in place with
no evidence of rejection. One subject rejected her skin graft after 11 months
following a total change of her antiretroviral therapy. Her viral load had become