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

32 Abstracts WeOrB1342-WePpB2092

XIV International AIDS Conference

WeOrBi342 Successful implementation in West African
laboratories of an alternative method to accurately
enumerate at lower cost circulating CD4+ T
lymphocytes in HIV-infected patients
S. Diacbougal, C. Chazallon2, M.D. Kazatchkine3, P. Van de Perre1,
A. Inwoley4, S. M'boup5, M. Prince David6, A. Tioro7, R. Soudr68,
J.P. Aboulker2, L. Weiss3. 'Centre Muraz, Centre Muraz, BP390,
Bobo-Dioulasso, Burkina Faso; 2INSERM SC10, Villejuif, France; 3H6pital
Europden Georges Pompidou, Paris, France; 4CEDRES, Abidjan, Cote divoire;
5Hdpital Le Dantec, Dakar, Senegal; 6CHU campus, Lome, Togo; 7INRSP,
Bamako, Mali; 8Faculte de Medecine, Ouagadougou, Burkina Faso
An international multicentric study was set to implement and validate a method
(DynabeadsR), alternative to high-tech reference method (flow cytometry, FC) for
measuring CD4 cell counts. This alternative method is based on CD4 T cell isolation using anti-CD4 mAB-coated magnetic beads.
Methods: 657 pairs of values of CD4 cell counts have been obtained by both
FC (TruCountR) and DynabeadsR from 684 blood samples of 301 HIV-infected
patients seen in one (n=112), two (n=61), three (n=75), four (n=40) or five (n=13)
occasions in 6 sites (Abidjan, Bamako, Bobo-Dioulasso, Dakar, Lome, Ouagadougou). Samples have been analyzed by 45 technicians. CD4 cell counts,
measured by FC, were below 200/mi in 40.5% of samples, between 200 and 350
in 19.5%, 350 and 500 in 9.4%, 500 and 1000 in 20% and above 1000 in 10.5%.
Results: The correlation coefficient between the 2 techniques was 0.89 (p<10 -4). The overall median difference between DynabeadsR and FC was -16 cells/ml
(p<10-4). Median difference, which was insignificant (+7.5 cells) for CD4 cell
counts below 200, increased, in a non-linear fashion, with CD4 levels. Thus, median differences were -23 and - 43.5 cells for CD4 cell counts between 200 and
350 and between 350 and 500, respectively. In 88.7% of cases, patients were
consistently classified at the threshold of 200 by both methods. Among 74 discrepant pairs of values, only 31 (4.7%, 95C1: 3.1-6.3) exhibited a difference of
more than 100 cells. Analyses of individual longitudinal data suggested that the
failing technique was DynabeadsR in 15 cases and FC in 11 cases.
Conclusions: DynabeadsR appeared to be a valid alternative to the reference
method. The ability to consistently classify at thresholds of CD4 cell counts relevant to clinical care was close to 95%. Finally, the implementation of this method in
6 sites involving 45 technicians was easy and successful. Our results encourage
the use of this alternative technique in countries with limited economic resources.
Presenting author: Serge Diagbouga, Centre Muraz, BP390, Bobo-Dioulasso,
Burkina Faso, Tel.: [phone], Fax: [phone], E-mail: diagbougaserge
@ hotmail.com
WeOrB1343 Development of affordable and portable HIV and
CD4 diagnostic tests using microchips
W.R. Rodriguez1, N. Christodoulides2, M. Ali2, M. Rodriguez2, P. Floriano2,
N. Bono2, B.D. Walker1, J.T. McDevitt2. IMassachusetts General Hospital,
Partners AIDS Research Center, Massachusetts General Hospital, 149 13th
Street, Rm 5234, Charlestown, MA 02129, United States; 2University of Texas,
Austin, United States
Background: Although the costs of HIV drugs have been reduced in parts of
the developing world, the costs of HIV laboratory tests remain high, and make
the establishment of treatment programs problematic. Affordable HIV diagnostics
that could be performed at regional and local health centers in developing countries are urgently needed, particularly CD4 counts and HIV RNA levels. We have
applied microchip technology to the development of affordable and practical HIV
diagnostic tests.
Methods: Agarose microbeads (<250 Itm diameter) were covalently coated with
molecules to detect HIV Ab, p24 Ag, HIV RNA and CD4 cells, and placed in
the wells of stamp-sized silicon microchips. Serum samples were processed on
the microchips, and HIV Ab, p24 Ag, HIV RNA or CD4 cells bound to the beads
were detected by transmitted light or fluorescent signals collected on a video CCD
chip interfaced with the detection chip. Signals were processed by image analysis
software contained in the detection device. Results were compared with standard
ELISA, PCR or flow cytometry-based assays.
Results: HIV Ab and p24 Ag in serum samples can be detected by this automated microchip system with sensitivity equal to or better than standard ELISA
assays.

Presenting author: William Rodriguez, Partners AIDS Research Center, Massachusetts General Hospital, 149 13th Street, Rm 5234, Charlestown, MA 02129,
United States, Tel.: [phone], Fax: [phone], E-mail: wrodriguez
@partners.org
WePpB2091 Liposomal doxorubicin (TLC D99, Myocet) in
combination with cyclophosphamide, vincristine
and prednisone is an active regimen for HIV
associated lymphoma
A.M. Levine1, A. Tulpulel, B.M. Espinal, R.S. Mocharnuk', D. Dharmapalal,
W.D. Boswell', L. Welles2. 1 USC Keck School of Medicine/Norris Cancer
Hospital, Los Angeles, CA, United States; 2Elan Corp, Princeton, United States
Background: TLC D-99 is a liposome-encapsulated formulation of doxorubicin,
with less cardiac toxicity than conventional doxorubicin, but retaining anti-tumor
activity. In this Phase 1/11 trial of CHOP-type chemotherapy, we substituted TLC
D-99 for conventional doxorubicin in patients (pts) with newly diagnosed HIVlymphoma.
Methods: Four dose levels of TLC-D99 were studied: 40, 50, 60, and 80 mg/m2
IV over one hour on day 1. For all levels, fixed doses of cyclophosphamide (750
mg/m2) and vincristine (1.4 mg/m2) were given IV on day 1; prednisone (100 mg)
was given orally days 1-5. Treatment was repeated every 21 days.
Results: 20 males and 1 female have been accrued; median age 41 (range 24 -61). The median CD4+ count was 115 /mm3, range (19-743); 16 (80%) pts had
a detectable HIV viral load. All pts were on combination antiretroviral therapy.
Pathologies included diffuse large cell in 16 pts; high grade in 2 pts; peripheral
T-cell, Burkitt, and Burkitt-like in 1 patient each. 15 (71%) had stage
IV disease. The main side effect of treatment was reversible grade 3 (24%) or
grade 4 (57%) neutropenia. Grade 3/4 anemia or thrombocytopenia was uncommon. Non-hematologic toxicities were primarily grade 1 or 2 in severity. No dose
limiting toxicity (grade 3 or 4 non-hematologic or prolonged hematologic) was
observed. Of 16 evaluable patients, 13 (81%) have attained complete remission
(CR) and 2 (13%) have had a partial remission (PR); 4 are too early in their treatment and 1 is inevaluable. The median duration of CR is not reached at 6.5+
months (range 1.7+-23.5+); to date 3 CRs have relapsed, all within 6 months of
CR. No significant difference in response rate was observed among the 3 dose
levels.
Conclusions: TLC D-99 substituted for conventional doxorubicin in the CHOP
regimen has significant activity in patients with HIV associated lymphoma, with
an overall response rate of 94%, and acceptable toxicity. Additional accrual is
ongoing.
Presenting author: Alexandra Levine, Norris Cancer Hospital, 1441 Eastlake
Avenue MS-34, Los Angeles, CA 90033, United States, Tel.: [phone],
Fax: [phone], E-mail: [email protected]
WePpB20921 Rituximab in HIV-related high-grade
non-Hodgkin's lymphomas: preliminary results of
an international multicenter pilot trial of rituximab
plus infusional cyclophosphamide, doxorubicin,
and etoposide (cde)
U. Tirellil, M. Spina, D. Bernardi1, E. Vaccher1, C. Simonelli, U. Jaeger2,
J.A. Sparano3. 'Division of Medical Oncology a cro, Aviano (pn), Italy;
2University Vienna, Austria; 3Montefiore medical center, New York, United States
Background: recent data suggest that the combination of rituximab plus
chemotherapy (ct) is more effective in the treatment of high grade non-hodgkin's
lymphoma (nhl). methods: with the aim to evaluate the efficacy and activity
of combining infusional cde plus rituximab, in june 1998 we started a phase
ii study using infusional cde (cyclophosphamide 187.5 mg/m2/day, doxorubicin
12.5 mg/m2/day and etoposide 60 mg/m2/day) administered by continuous intravenous infusion for 4 days every 4 weeks for up to 6 cycles and rituximab 375
mg/m2 i.v. on day 1 prior to each. highly active antiretroviral therapy (haart) was
given concomitantly with ct, irrespectively of cd4+ cell count and hiv viral load.
results: from june 1998 to october 2001 41 patients have been enrolled and 38
patients are evaluable for response and toxicity, thirty-five patients (85%) were
male and median age was 38 years (range 29-65). the median cd4+ cell count
was 120 (range 3-578) and the median performance status (ecog) was 1 (range
0-2). sixty-one percent of patients had advanced stage (iii-iv) disease and 49%
had b symptoms. twenty-nine out of 38 patients (76%) achieved a complete remission (cr), 2/38 (5%) had a partial remission and 7 patients progressed. only 3
patients out of 29 crs have relapsed and 32/41 patients are alive, the major cause
of death was nhl (8/9 patients). grade 3-4 neutropenia, anemia and thrombocytopenia were observed in 80%, 37% and 29% of patients respectively, twentynine percent of patients developped bacterial infections during neutropenia no
toxic deaths were observed, with a median follow-up of 12 months, the actuarial overal survival and progression-free survival at 2 years were 70% and 86%
respectively, conclusions: the combination of rituximab and infusional cde in pa
tients with hiv-associtated nhl is safe and feasible, even if there is an increase
in bacterial infections, however, all infections were cured with antibiotics and no
toxic deaths were observed.
Presenting author: Umberto Tirelli, Division of medical oncology a, National
Cancer Institute, via pedemontana occidentale 12, 33081- aviano (pn), Italy, Tel.:
[phone], Fax: [phone], E-mail: [email protected]

Fig 1 (left). CD4 cells bound to beads detected by a) transmitted light or b) fluorescence.
Fig 2 (right). Serum containing both HBV Ab and HIV-1 Ab at titers of 1:200 detected on a
microchip.
Preliminary studies suggest that microchip-based assays for HIV RNA detection
and CD4 cell count are also feasible.
Conclusions: Microchip-based detection of HIV Ab, p24 Ag, RNA and CD4
counts are feasible. These tests may be combined on a single disposable and
inexpensive microchip. A battery-powered, hand-held chip reader has also been
developed. Microchip-based HIV diagnostics could make affordable laboratory
testing in resource-poor settings a possibility.