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

12th World AIDS Conference

Abstracts 31160-31164 527

31160 Effect of prolonged HAART on immune function and on
viral load in lymph tissue
Royt Steigbigel, E. Irwin, B. Craddock, E. Govek, M. Burk, S. Morrison. Division
of Infectious Diseases HSC Suny SB, Stony Brook NY 11794-8153, USA
Background: Highly active antiretroviral therapy (HAART) in many patients (pts)
results in undetectable viral load in blood for years. Its effects on restoration of
immune function, neutralizing antibody (NAb), and viral load in lymph tissue, is
incompletely known.
Methods: In a group of 16 pts receiving HAART for >1 y. (median 126 wks.)
we studied over time (1) hypergammaglobulinemia (HyG, an early abnormality in
pts with HIV infection); (2) levels of anti-gp120, potentially NAb; and (3) viral load
in whole lymph node (LN) biopsies by co-culture, bDNA analysis of HIV RNA,
and proviral DNA by PCR in situ.
Results: Sixteen pts, median viral load 45,860 copies/ml before therapy, received up to 144 wks. of indinavir with RT inhibitors. Fourteen of 16 pts tested
had HyG (>9 mg IgG/ml by ELISA) initially and it remained in all 9 HyG pts
who developed undetectable viral load in blood ("Responders," R), and in all 5
HyG nonR pts. However, in those with detectable anti-gp120 Ab, Ab declined
significantly (generally noticeable after 0.5 y.) in 4/7 R in whom it was measured
thus far, but in only 1/3 nonR. The decrease in anti-gpl20 appeared specific,
as anti-tetanus Ab was unchanged. LN viral load declined in parallel with that in
blood, but was always higher; proviral DNA at 30 wk. did not decrease in LN cells.
Two R (i.e. < 20 copies RNA/ml) had proviral DNA in -0.007% and ~0.002%
of CD4+ LN cells at 2 y. 10 mo. and 2 y. 7 mo., respectively, whereas none was
detected in their blood cells. Virus was grown from LN of one of these pts. It had
wild-type protease but multiple mutations in RT.
Conclusions: In pts responsive to HAART HyG persists even after 2 y. There
is evidence of proviral DNA in lymph tissue after 2.5 y. There is a decline in
potentially NAb with return of naive CD4 cells. Therefore, immunization with HIV
antigens should be attempted to see if it elicits immune control of residual HIV.
31161 Anti-chemokine antibodies in sera of HIV-1-infected
patients
Caner SOsal', V. Daniel1, S. Kissler', C. Uhle2, R. Zimmermann2, C. Opelz1.
SInst. of Immuno. Univ of Heidelberg INF 305 D-69120 Heidelberg; 2Kurpfalz
Hospital Heidelberg, Germany
Objective: To investigate whether antibodies against the /-chemokines MIP-lo
and RANTES and against the u-chemokine SDF-lu occur in sera of HIV-1-infected
patients and whether they play a role in disease progression.
Methods: Sera of HIV+ hemophilia patients and healthy controls were tested
for IgG-anti-chemokine activities in ELISA. Results are given as mean optical
density (OD) ~ SEM.
Results: When positivity was defined as > mean OD+2SD of healthy controls, 80% of the HIV+ sera were positive for anti-MIP-1, 58% for anti-RANTES,
and 50% for anti-SDF-1. 318 sera of 44 HIV+ patients had significantly higher
anti-MIP-1 (294 ~ 12), anti-RANTES (245 ~ 10), and anti-SDF-1 activities
(322 ~ 13) than sera of 35 healthy individuals (91 ~ 5, 105 ~ 6, and 130
~ 11; p - 0.0001 in all cases). Significant associations were observed between the 3 anti-chemokine activities (anti-MIP-1/anti-RANTES: r = 0.9; antiMIP-1/anti-SDF-1: r = 0.6; anti-SDF-1/anti-RANTES: 0.5; p < 0.0001). Although
the viral load was strikingly higher in patients with <50 CD4 cells//pl than in
patients with >50 CD4 cells/pl, significantly lower anti-MIP-1 and anti-RANTES
were found in 100 sera of patients with <50 CD4 cells//il than in 218 sera of
patients with:>50 CD4 cells//l (p = 0.001 for both), suggesting that disappearance
of M-tropic virus in advanced disease causes a decrease in anti-/l-chemokine
activity. Conversely, anti-SDF-1 was still high in late stage disease and correlated
with viral load (r = 0.25, p = 0.002). Especially in patients with >200 CD4 cells//l,
anti-MIP-1 and anti-RANTES correlated with CD8 counts (r = 0.4 and 0.35),
whereas anti-SDF1 did not. In patients with a high CD4 count of >500//l1, anti-/I
-chemokine activities were associated strikingly with anti-V3-loop activity (r = 0.7
and 0.8) and, emphasizing the importance of /-chemokines in early disease,
inversely with CD4 counts (r = -0.39 and -0.33). Anti-RANTES antibodies that
were isolated from sera of 5 different patients crossreacted with gp120 V3-loop
peptide-coated plates, and an affinity-purified anti-V3-loop antibody bound to
MIP-1-and RANTES-coated plates.
Conclusions: Our data indicate that anti-chemokine activities in sera of HIV+
patients are due to a crossreaction of anti-V3-loop antibodies with chemokines.
Anti-chemokine activity may play a deleterious role in AIDS pathogenesis by reducing the chemokines' potential to inhibit HIV-1 entry into CD4+ cells, especially
in early disease.
S31162 Anti-lymphocyte antibodies and viral load in AIDS
patients
leda M. Longon-Maugeri1, H.A. Harima', S.G. Oliveira', C.F.H. Granato2,
Z.F. Peixinhol, N.F. Mendes'. 1Rua Botucatu 862 4~ Andar, 04023-900 Sao

Paulo, Universidade Federal de Sao Paulo, Sao Paulo, SP; 2Laboratorio Anal.
Pesq. Gastao Fleury SC, Sao Paulo, SP Brasil
Objective: To investigate the presence of anti-lymphocyte antibodies and viral
load in AIDS patients. To standardize immunoenzymatic assays to detect these
antibodies and compare with microlymphocytotoxicity test.
Design: Retrospective, controlled study.

Methods: Serum samples of 19 AIDS patients were analysed for the presence
of anti-lymphocyte antibodies by microlymphocytotoxicity (MLCT) and immunoenzimatic assay using as antigen whole lymphocytes (CELISA) or supernatant from
lysate lymphocytes obtained by ultracentrifugation (MEM-ELISA). Lymphocyte
panel from 14 normal individuals was employed in both assays. Amplicor HIV-1
(ROCHE) was used to investigate viral load.
Results: 8/19 (42.1%) patients presented a viral load >100,000 copies, 7/19
(36.8%) between 10,000 to 100,000 copies and 4/19 (21%) <10,000 copies.
Thirteen sera were reactive (>50% of lysis of the target cells) with at least one
cell of the panel, by MLCT. Five of them (26.3%) were reactive with 50% to
100% of panel and presented a viral load over than 30,000 copies. The cytotoxic
anti-lymphocyte antibodies belong to the IgM class, since dithiothreitol treated
sera abolished the reaction. We standardized the CELISA and MEM-ELISA determining the ideal concentration and fixation of lymphocytes and/or their lysate
in the solid phase. Cut-off was established per plate = X HIV-1+ controls + X
HIV-1 controls x 0.15. In CELISA all sera were reactive with at least one cell
of the panel. Thirteen sera (68.4%) reacted with 50% to 100% against the cells
of the panel. The viral load of these sera was -7,400 copies. By MEM-ELISA,
anti-lymphocyte antibodies were detected in 9 (47.4%) against 50% to 100% of
the panel. The viral load of these sera was >7,400 copies.
Conclusion: The presence of anti-lymphocyte antibodies seems to be one of
the mechanisms of lymphocyte destruction in AIDS patients. We observed that
both CELISA and MEM-ELISA were more sensitive than microlymphocytotoxicity.
31163 Study of cellular and humoral immune factors in
colostrum of HIV infected and non-infected lactating
mothers
Manita Williamson', Rashid H. Merchant2, A.A. Manohar3, H.A. Kamat3,
G.V. Koppikar3. 1N. Wadia Mat. Hosp. (NWMH)-Div. Neonat. Acharya Dhonde
Marg, Parel, Mumbai-400012; 2Prog. Dir. -Perinatal HIV Center (NWMH)
Mumbai; 3T.N. Medical College & BYL NAIR Hosp. Mumbai, India
Background: This study was done to quantify cellular & humoral immune factors
in colostrum of HIV seropositive (Study group) & seronegative lactating women
(Control group).
Methods: 15 ml. colostrum was collected from 130 asymptomatic women 62
(Study) & 68 (Controls) in two sterile siliconised glass test tubes. The total cell
count, cell viability, differential count, T cell count by E. rosette assay and phagocytic activity of macrophages as well as the IgA, IgM, IgG levels of colostrum
were estimated in all.
Results: The percentage phagocytosis and percentage T cell number in the
study group was 57.3 ~ 7.1 & 37.5 ~ 5.4 as compared to 70.3 ~ 7.1 & 50.5 ~
3.1 in controls (p < 0.001). The IgA & IgG content in the study group were 241.4
~ 110.3 & 9.6 ~ 7.0 as compared to 289.3 ~ 129.1 & 12.6 ~ 7.9 in controls (p
< 0.05). There was a statistically significant different in the phagocytosis, T cell
number, as also the IgA & IgG contents in the two groups.
Conclusion; Results of this study indicate decrease in some cellular & humoral
defence factors in colostrum of HIV seropositive women compared to seronegative ones. Whether these reduce the overall beneficial effects of breast milk is a
moot question. The synergistic effect of not breast feeding & poor bottle hygeine
can be disastrous in developing countries.
S31164 Role of mannon-biding protein (MBP) in infectious
episodes in pediatric AIDS
Yu Ching Lian', M. Della Negra', A.S. Grumach2, M. Kirschfink5, H.M. Yang4,
W. Queirozl. 'Instituto de Infectologia Emilio Ribas, Rua Pamplona 356, 11
Andar, Sao Paulo; 2lnst. Crianga H.C.F.M.J.S.FP, So Paulo; 4 Unicamp IMECC
DMA, Campinas, SP, Brazil; 5lnst. Immunol. of Heidelberg University,
Heidelberg, Germany
Objective: The aim of this study is to analyze the value of Mannon-binding
Protein (MBP) and possible correlation with clinical manifestations presented in
each stage of HIV infection in the pediatric population.
Methods: One hundred twenty seven HIV infected, vertically exposed children
were studied. Their HIV diagnosis and classification followed the current CDC
criteria. MBP dosage (ELISA-Nunc Maxisorp F 96) and CD4/CD8 cell count (flow
cytometry) were correlated with infectious and clinical manifestations. Statistical
analysis was performed using ANOVA (parametric) and Kuskal-Wallis (non-parametric) tests.
Results: Sixty five (51.2%) were female and 62 (48.8%) were male. Age ranged
from 11 to 134 months (mean 59.2 mo.). The most frequent infectious episodes
were bacterial pneumonia (44.5%), otitis media (33.6%), tuberculosis (32.0%),
diarrheal illness (20.3%), PCP (9.4%). We did not observed any statistically
significant relationship between MBP values and clinical categories or CD4 cell
count.
N1 N2 N3 Al A2 A3 B1 B2 B3 C1 C2 C3 P
# 9 5 2 6 10 5 12 13 11 14 16 24

Mean Age 51.3 80.4 53.0 65.5 72.1 62.2 53.1 55.4 48.2 58.5 53.8 64.8 0.62
CD4 1322 478 420 1022 458 279 1327 541 334 1174 580 208 0.000
CD8 2059 1693 396 1547 1506 1023 1503 2199 1615 1978 2544 1207 0.02
MBP 4.34 1.65 3.45 4.52 3.41 5.87 3.60 3.43 4.18 5.11 6.99 5.73 0.66
Obs.: Reference values for MBP: up to 10/ g/ml.