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

532 Abstracts 31183-31187

12th World AIDS Conference

undetectable and her CD4 cells had raisen to 211. The two remaining subjects
are still alive 19 months later with no sign of skin rejection. No important betterment of surrogate markers were seen in subjects who tolerate there skin
grafts.
Conclusion: Major improvement of surrogate markers after changing antiretroviral regimen suggests that HIV immunoincompetent subject could restore their
immune status since a skin allograft properly applied cannot be retained except
in T-cell important immunodeficiency. As far as we know, it is the first time that
tissue allograft is used to assess T-cell dysfunction in HIV infected patients.
31183 Immunohaematological reference ranges for adult HIV
Ethiopians: Effects of HIV-1 infection
Aster Tsegaye1, T. Mesele1, T. Tilahun1, E. Hailu1, R. Doorly1, A. Fontanet1,
T. Rinke de Wit1. Ethiopian Netherlands AIDS Research Project/ENARP Addis
Abeba; 2EHNRI, Addis Abeba, Ethiopia
Objectives: Establishment of immuno-haematological reference ranges for adult
HIV-Ethiopians and comparison with adult HIV+ Ethiopians.
Design: Cross sectional study.
Methods: For haematology, Coulter Counter measurements on whole blood
of Hb, Hct, RBC, Pit and WBC were performed on 635 apparently healthy HIVEthiopians (346 M/289 F) and 89 HIV+ Ethiopians (46 M/43 F), who participated in
a cohort study on the progression of HIV-1 infection in Ethiopia. For immunology,
FACScan measurements on whole blood for various lymphocyte subsets were
performed on 113 HIV- (62 M/51 F) and 83 HIV+ (43 M/40 F) Ethiopian cohort
participants. Results were compared to healthy Dutch blood donor controls.
Results: Immuno-haematological 95% percentile ranges for healthy adult HIVEthiopians were established. Gender specific differences, independent of HIV
status, were detected for CD4+ T-cells, RBC, Hb and HCT. Immunological 95%
percentile ranges were: lymphocytes 1042-3089/ul, T-cells 754-2412/ul, CD4+
T-cells 366-1315/ul, CD8+ T-cells 312-1421/ul, CD4/CD8 ratio's 0.5-2.5, B-cells
49-401/ul and NK cells 83-635/ul. The absolute CD4+ T-cell counts of healthy
Ethiopians (mean 770; median 753) are significantly lower than of European
controls (mean 1066, median 980), while the opposite is true for CD8+ T-cells
(Ethiopia: mean 714, median 637; Europe: mean 528, median 500). In HIV+
Ethiopians the following parameters changed significantly compared to HIV- individuals: decrease of CD4+ T-cells, lymphocytes, Hb, HCT, RBC, WBC and
increase of CD8+ T-cells. In 31 HIV+ individuals with follow up values, the rate of
CD4 and Hb declines over 1 year were 95 cells/ul and 1 g/dl respectively.
Conclusion: Immuno-haematological reference values have been established
for healthy Ethiopian adults, which will be of use for interpreting changes in these
values in HIV infected Ethiopians during progression of the disease.
31184 Cell surface markers of immune activation in dual
infection with HIV and Mycobacterium tuberculosis
Lalit Dar1, G.K. Singh1, P. Jain2, R. Kumar2, S. Brook1, J.P. Walil, P. Seth1.
1 Department of Microbiology, A....M.S., Ansari Nagar, New Delhi 110029;
2Haematology IRCH AIIMS, Ansari Nagar, India
Objectives: Flow cytometric comparison of lymphocytic immune activation markers in persons with HIV infection alone and persons with dual infection with HIV
and Mycobacterium tuberculosis.
Methods: Expression of CD38 and CD45RO, on CD8 and CD4 lymphocytes
respectively, was analysed by flow cytometry in 15 persons with dual infection due
to HIV and M. tuberculosis, 20 persons with HIV infection alone (asymptomatic
and symptomatic) and 20 matched healthy controls. Whole blood was collected
in EDTA. Fluorescent staining was carried out with FITC and PE conjugated
monoclonal antibodies to CD4, CD8, CD38 and CD45RO (Becton Dickinson);
gifted by Fogarty program through Dr. J. L. Fahey, Director, CIRID, UCLA, USA.
Whole blood samples were stained and lysed by single step commercial lysing
solution (Optilyse C, Immunotech) and analysed by two-color flow cytometry on
a Coulter Elite flow cytometer.
Results: The percentage of CD4 positive cells decreased from a mean of 36%
in healthy controls to 15.4% in persons with HIV infection and 9.7% in persons
with dual infection (with HIV and M. tuberculosis). Mean percentage of CD8 positive cells decreased from 22.1% to 44.3% and 39.4% in these study groups. The
percentage of activated CD8+ cells (i.e. CD38+CD8+/CD8+) also increased from
18.7% in control group to 54.6% in HIV infection. A further increase in activated
CD8+ lymphocytes to 73.4% was demonstrable in dual infection, as evidenced by
this phenotypic marker of immune activation. The percentage of activated CD4+
cells (i.e. CD45RO+CD4+/CD4+) decreased from a mean of 41.5% in the healthy
control group to 23.3% in HIV infected persons, but showed an increase to a
mean of 40.8% in dual infection.
Conclusions: In HIV infection alone, a decrease was seen in activated CD4+
cells (based on expression of cell phenotypic marker of immune activation,
CD45RO) as compared to healthy controls. In dual infection this increased to
levels comparable to healthy controls. Activated CD8+ cells (based on CD38
expression), increased in HIV infected patients from the normal levels seen in
healthy controls, and showed a further increase in dual infection.

31185 rlL-2 rescues in vitro functions of lymphocytes from
HIV-infected individuals
Pia Afzelius, Jo Nielsen, Jes Hansen. Dept. of Infectious Dis. 144, Huidoure
Hospital, 2650 Huidoure, Denmark
Introduction: CD4+ lymphocyte counts and helper functions decline during HIVinfection as assessed by lymphocyte proliferation and IL-2 production as does
IL-2R expressions. rlL-2 has recently been shown to be effective in increasing CD4+ lymphocyte counts in HIV-infected individuals, and has also been
demonstrated to improve functionality of PBMC from the patients.
Methods: PBMC from eight HIV-seropositive individuals and eight healthy
blood donors were treated in vitro for 14 days with IL-2 plus PHA day 0-3. The
cells were studied before and after treatment for mRNA expression of IL-2 and
IL-2R by RT-PCR. The proliferative capacities as responses to mitogens were
studied in a standard proliferation assay. FACS analyses were performed for
activation markers and clonality. HIV-antigen was measured in supernatants from
PBMC of HIV-seropositive individuals by ELISA.
Results: Without rlL-2 the proliferation responses of patients were significantly
lower than those of donors (p = 0.002), whereas responses to PHA + rlL-2 were
comparable. Expressions of CD25 on CD4+ T cells tended to be higher and were
higher on CD8+ T cells in patients than in donors. Prior to IL-2 treatment, the
ability to respond to PHA with increased mRNA expressions of IL-2 and IL-2R was
lower in patients than in donors (p = 0.02 and p = 0.05). After IL-2 treatment the
ability to respond with mRNA expression of IL-2 and IL-2R was improved to the
levels of donors. The responses of patient cells to PHA with mRNA expression
of IL-2R correlated with the ability to increase the CD25 percentage on PBMC
(p = 0.007) after IL-2 treatment. The amount of HIV-antigen of infected cells was
unchanged.
Conclusion: These findings suggest existence of a reversible IL-2 and IL-2R
defect at the pretranscriptional or transcriptional level in PBMC from HIV-infected
individuals. This may partly explain T lymphocyte anergy during HIV-infection.
338*/31186 Immunologic function and reconstitution in
HIV-infected patients on highly active antiretroviral
therapy (HAART)
Ilana Fogelman1, V.J. Davey2, M. Sneller2, M.B. Feinberg3, H.D. Ochs4,
J.P. Siegel1, H.C. Lane2. 1FDA 1401 Rockville Pike HFM 570, Rockville, MD
20852; 2NIAID, Bethesda; 30AR-NIH, Bethesda; 4University of Washington,
Seattle, WA, USA
Objectives: To evaluate the effects of HAART and disease stage on immune
function and reconstitution in HIV-infected patients.
Design & Methods: Open label, controlled study. 16 HIV-infected patients and
8 controls received three immunizations with the neo-antigen bacteriophage phi
X174, six weeks apart. Since memory amplification of antibody (Ab) titers with
repeated immunizations and antibody isotype switching (AblS) from IgM to IgG
require adequate T cell function, these parameters were used as indicators of T
cell function in vivo.
Results: 3 types of Ab responses were identified: I. Ab titers that reached the
normal range. II. Ab titers that increased with repeated immunizations, but were
below the normal range. III. Ab titers that decreased with repeated immunizations.
5 patients had a type I, 4 had a type II and 7 had a type III response. All type I
and II patients had a CD4 count >200 cells/uL. All type I and II patients had a
viral load (VL) < 500 copies/mL, except for a type I long-term non-progressor not
on therapy and a type II patient who switched to HAART two weeks after primary
immunization and then developed a VL < 500. Within type III, 4 patients had a
CD4 count <200; two with VL > 500, one with VL < 500 and one who received
HAART two weeks after primary immunization and then developed a VL < 500.
Type III also included 2 patients with CD4 counts of 471 and 291 and VL > 500,
and 1 patient with a CD4 count of 892 and VL < 500. The proportion of IgG in
tertiary responses was 45% for type I, 32% for type II and 4% for type III (normal
range is >75%).
Conclusion: Higher CD4 counts and lower viral load were associated with a
better immune response to bacteriophage phi X174 immunization and evidence
of T cell function in vivo. However, the majority of patients in our cohort failed
to mount a totally normal immune response to this neo-antigen, despite viral
suppression sustained for several months and normal CD4 counts in several
instances.
31187 Enhanced of phagocytosis and germicidal activity in
originating neutrophils and monocytes from bone
marrow cells cultures of AIDS patients
Carlos Sergio Barragan, G.J. Deluchi, A.M. Cahizal, J. Benetucci. Fundai Htal
Muniz Uspallata 2272 CPI282, Rosario 294 11B (23) CP1424 Buenos Aires
Capital Federal, Buenos Aires, Argentina
Objetives: To study originating neutrophils and monocytes from bone marrow
stem cells culture with recombinant IL-3 and GM-CSF of AIDS patients.

Methods: All patients were classified as belonging to group 3C according to
the CDC classification. Funtional activity of polymorphonuclear neutrophils and
monocytes from peripheral blood was tested in this HIV-1 infected patients. Ten
milliliters of bone marrow fluid was aspirated from esternon into an heparinized
syringe. In 22 cases primary cell culture of bone marrow were performed as
follow: