Final Program and Oral Abstracts [International Conference on AIDS (8th: 1992: Amsterdam, Netherlands)]

MoA 0007-MoA 0012

MONDAY, 20 JULY 1992

MoA 0007 ANUID=CYE-OLONYSTIMUIATING FACIOR
MoA 0007 -CSF IN AIDS-PATIENIS WITH LEUKOCYTES < 1.0 x 109/L.
Bratt G6ran, Griitzmeier S, Lund B, Sandstr6m E.
Deparuttnt of Dermatovenerology, South Hospital, S-118 83 STOCKHOIM
Issue: AIDS-patients, that are treated with chemotherapy for Kaposis
sarkoma and Ganciclovir/DHPG) for CMV-retinitis often get severe neutropenia, that
limits the treatment and increasis the incidense of bacterial infections. Can treatment with G-CSF increase white blood cell count WBC, lcoer the incidense of bacterial
infections in these patients? Which dose of G-CSF is optimal in these patients?
Methods: 17 AIDS-patients (B with KS, 8 with CMV-infection and one with malignant
lymphcma) and WBC < 1.0 x 10 /L. have been treated with G-CSF 5g/kAg s c. 6 were
pilot cases and 11 (6 with KS and 5 with CMV-retinitis) are in a prospektive study
to evaluate the efficasy of intermittant G-CSF therapy. In the prospektive study
G-CSF was administered 5ug/kg until WBC < 1.5 x 109/L. WBC and differential count
were followed every 12 hours the first 72 hours and then daily the first week. A new
dose of G-CSF was given as soon as WBC was < 1.5 x 109/L.
Results: In the 6 pilot patients WBC increased to 9.46 x 109/L (M) (0.2-17.1 x 109/L)
on the third day after daily injections of G-CSF. All 11 patiens in the prospektive
study had an increase of neutrophile granulocytes to > 1.0 x 10 /L within 24 hours.
intenance treatment varied between agA/kg daily and 5e/g/kg every 14 days. Two patients had transient skeletal pain but no other side effekts were seen. Most patients
learned to inject G-CSF themselves. No bacterial infections were seen when WBC was
> 1.5 x 109/L.or neutrophils were > 1.0 x 109/L. Follow up time was 7 month (M).
Conclusion: G-CSF causes a rapid increase in WBC and neutrophile count in AIDS-patients with neutropenia. Intermittant G-CSF seems to increase neutrophils to a level
with a low risk of infections and thus makes it possible to continue chemotherapy
and ganciclovir treatment in these patients.
MoA 0009 Specificity of antibodies produced against native or desialylated
recombinant HIV-1 gp160. Benjouad. Abdelaziz*:; Gluckman,J.C. ";
Rochat, HW; Montagnier, L."'; Bahraoui, E'.*'CNRS URA 1455, Marseille,
*"CNRS URA 1463, CERVI, H6pital de la Pitid-Salpdtrirre, Paris, "'Unitd
d'oncologie virale, Institut Pasteur, Paris, France.
QOiective: In contrast to antibodies produced against native or fully deglycosylated HIV-1 gp160 in rabbits,
antibodies raised against desialylated HIV-1 gp160 recognized also gp140 from HIV-2 at high titers. The
population of antibodies responsible for this cross-reactivity was further analyzed.
Methods: Rabbits were immunized with native or desialylated gpl60. Sera were analyzed by ELISA and by
RIA against HIV-1, HIV-2 and SIV envelope glycoproteins. Synthetic peptides were used to map the regions
recognized by these antibodies.
Results: While all sera from immunized rabbits recognized HIV-1 gp160, only antibodies elicited by
desialylated gp160 cross-reacted with HIV-2 gp140 at high titers, even when it was fully deglycosylated.
ELISA analysis against a panel of synthetic peptides showed that anti-desialylated gp160 antibodies
recognized a P41 synthetic peptide (residues 252 to 266) in addition to the reactivity pattem obtained with
anti-native gp160 antibodies. Inhibition assays using synthetic peptides as competitors showed that crossreactivity with gpl40 was due to an antibody population specific to the region encompassing the HIV-1 gp41
immunodominant epitope mimicked by peptide P39 (aa: 583-610), the latter being able to totally inhibit the
formation of complexes between HIV-2 radiolabled gp140 and antibodies against desialylated HIV-1 gp160
antibodies. This was al the more surprinsing that both anti-native and anti-desiatylated gp160 antibodies
recognized peptide P39 with the same titers. In addition, when anti-desialylated gp160 was adsorbed on
P39 affinity column, the retained antibodies still bound HIV-2 gp140. Interstingly, this cross-reactive antibody
population did not recognize SIV gpl40 which share more than 90% homology with HIV-2 gp140 within this
region.
Conclusion: Cross-reactivity with HIV-2 gpl40 of antibodies elicited by desialylated HIV-1 gpl60 illustrates
the influence of carbohydrate moieties on the specificity of produced antibodies. Moreover, this crossreactivity could explain the positive reactivity observed, against both HIV-1 and HIV-2, in some human sera.

MOA 0008 (r-HuEO) TREATMENT IND RECOMBINANT HUMAN ERYTHROPOIETIN
MoA 0008 (r-HuEPO) TREATMENT IND PROTOCOL FOR THE ANEMIA OF AIDS - OVERALL
RESULTS AND AZT SUBGROUP ANALYSIS. Phair, John*; Abels, R.**
*Northwestern University Medical School, Chicago, IL, USA, **The
R.W. Johnson Pharmaceutical Research Institute, Raritan, NJ, USA
Oblectives: Anemia associated with HIV infection may be due to reduced erythropoiesis
related to the disease itself or to concomitant medications (e.g., zidovudine [AZT]).
Clinical studies have shown r-HuEPO to be effective in correcting the anemia of AZTtreated HIV-infected patients with baseline erythropoietin (EPO) levels <500 mU/mL.
Results of a Treatment IND (T-IND) protocol which provided r-HuEPO to approximately
2,000 anemic AIDS patients are presented here.
Methods: Enrollment criteria included a clinical diagnosis of AIDS, serum EPO level
<500 mU/mL, hematocrit (HCT) <30%, and age >12 years. Initial r-HuEPO dosage was
4,000 Units (U) subcutaneously (sc) for 6 days each week. This could be increased
sequentially to 8,000 U sc for 6 days per week.
Results: Overall, 40% of patients (N=1,943) required at least 1 transfusion in the
6 weeks prior to study entry (baseline). Corresponding percentages were 22% (N=1,387)
and 18% (N=650) after 12 and 24 weeks of r-HuEPO treatment. HCT levels were 28.0% at
baseline, 33.1% at week 12, and 33.8% at week 24. Response to therapy, defined as an
increase from baseline in HCT of >6 percentage points with no transfusion 28 days
prior to achieving that HCT, was observed in 44% of patients. Improved quality of
life with respect to increased energy and reduced health distress was noted after 24
weeks in a subset of patients completing quality of life questionnaires. Correction
of anemia by r-HuEPO was also observed in a subset of 671 patients who received
concomitant AZT therapy for at least 75% of their participation in the T-IND.
Transfusion requirement decreased from 40% at baseline to 24% at week 12 (N=553) and
19% at week 24 (N=277). HCT increased from 28.7% at baseline to 33.1% at week 12 and
33.8% at week 24. Response to therapy was noted in 46% of patients. The efficacy of
r-HuEPO was unrelated to reductions in AZT dosage. Adverse experiences not clearly
related to AIDS were reported by 11% of patients.
conclusions: Overall, in a study population of 1,943 AIDS patients, r-HuEPO therapy
corrected anemia and was well tolerated. Results with respect to increased HCT and
decreased transfusion requirements were comparable in a subgroup of patients who had
received AZT for at least 75% of the duration of r-HuEPO treatment.
John P. Phair, M.D., Northwestern University Medical School, 680
N. Lake Shore Drive, Suite 1106, Chicago, IL 60611-4402, USA
Telephone: [phone]; FAX: [phone]
M oA 0010 POTENTIAL OF SELECTIVELY DEGLYCOSYLATED HIV-1 GP120 AS A MORE
EFFECTIVE VACCINE ANTIGEN
Lee. Tun-Hou. Lee, C.N., Lee, W.R., Cheng, Y. L.,in, iH. L. and Essex, M.
Harvard School of Public Health, Boston, Massachusetts 02115 USA
Obiectives: Immune decoy mediated by carbohydrate moieties of viral envelop proteins was observed
previously in some retroviruses. HIV-1 gp120 is a heavily glycosylated protein with approximately half of
its molecular mass contributed by N-linked sugars. One prediction of the hypothesis that certain N-linked
sugars of gpl20 may serve as immune decoys is that the removal of these N-linked sugars will have no
effect on in vitro replication of HIV-1. The objective of this study is to evaluate our prediction that HIV-1
gp120 contains carbohydrate moieties that are dispensible for in vitro replication of HIV-1.
Methods: N-linked glycosylation mutants of HIV-1 were constructed by site-directed mutagenesis. The
infectivity of these mutants was assayed on CD4-positive cells.
Results: Dispensible N-linked glycosylation sites are found to be clustered in the C-terminal region of
gpl20, which contains some of the putative CD4-binding sites. The removal of as many as seven of the
ten N-linked sites located between the V3 region and the C-terminus of gpl20 did not affect the
processing of the gpl60 precusor, nor did it abolish viral infectivity. In contrast, the removal of two Nlinked sites at some positions in the N-terminal half of the gp120 was sufficient to abolish viral infectivity.
Conclusions: Our data showed that a large number of N-linked sites located in the functionally important
region of gp120 were not required for viral replication. This finding is compatible with the hypothesis that
some of the N-linked sugars of gpl20 may be evolutionarily conserved to evade a host immune
response. A prediction of our finding is that selectively deglycosylated gpl20 may represent a more
effective vaccine antigen than the fully glycosylated gp120. In this regard, it should be noted that
asymptomatic HIV carriers lacking gpl20 antibodies to an unglycosylated reconibinant peptide covering
the C-terminal 170 amino acid residues was previously found to be more likely to progress to AIDS than
those with such antibodies. Taken together, our finding raise the possibility that selectively
deglycosylated gp120 may represent a more effective vaccine antigen than the fully glycosylated gp120.

Benjouad, Abdelaziz, Laboratoire de Biochimie Facultf de Medecine Nord, Bd. P.
Dramard 13015 Marseille France, Telephone [phone], Fax [phone].
VACCINE-THERAPY WITH GP160: EFFECT ON
HUMAN ANTI-GP120 ANTIBODY SPECTROTYPE.
Biselli. Roberto*; Loomis, L.D.^; Del Bono, V.*; Burke, D.S.*; Redfield, R.R.'; Birx, D.L.*
*Department of Retroviral Research, Walter Reed Army Institute of Research, and ^Retroviral
Research Laboratory, Henry M. Jackson Foundation, Rockville, MD 20850

MoA 0011

OBJECTIVES. Infection with HIV type 1 causes a progressive failure of the immune system
and, among other things, a decline in the number and/or function of B-cell clones recruited
in specific humoral responses. Spectrotypic analysis, as obtained by Isoelectric Focusing (IEF)
and reverse blotting, is one technique for indirectly evaluating the activity and the number
of specific B-cell clones. The aim of this study has been to evaluate by IEF the modulation of
the anti-gp120 specific B-cell clones generated by post-infection vaccination with
recombinant gpl60. This method allows specifically for measurement of conformationally
intact epitopes.
METHODS. We studied the anti-HIVI(LAI)gpl20 spectrotype in 30 volunteer HIV-infected
patients in early stage (Walter Reed stage 1 or 2) of the disease, undergoing vaccine therapy
with gpl60 (LAI, from baculovirus, MicroGeneSys), and in 8 patients in a blinded matched
cohort, receiving a placebo vaccine. The duration of the follow-up evaluation was 21 months.
RESULTS. Twenty-five out of the 30 vaccinated patients displayed a clear banding pattern in
IEF: the spectrotype was basically oligoclonal, with bands mainly in alkaline and neutral pH
range. Of the reacting subjects, 7 (28%) showed the same pattern in all samples, while 10
(40%) displayed the same spectrotype but with a progressive increase in band intensity
during the course of immunization. Finally, the last 8 patients (32%) showed both an increase
in band intensity and number of bands. Sera from the 8 patients receiving placebo vaccine
showed no change in band intensity and no new bands were seen.
CONCLUSION. In the present study we demontrated that a vaccine therapy with recombinant
gpl60 has been able to induce an enlargement of the anti-HIVI(LAI)gpl20 B-cell clone pool
in 8 out of the 30 HIV-infected patients. In additional 10 subjects, the spectrotype showed an
increase in band intensity, suggesting a raise in antibody synthesis by B-cell clones already
recruited for the specific immune response. This is in sharp contrast with previous studies
showing only decreases in bands during the long course of the disease.
Biselli.Rober to Aeronautica Mili o DASRS.Re Me9icina, Aeroporto ratIca 01 mare_, uu omezai Ifoma),
ITALIA. Tel:..-396-9108119, FAX:..-396-9124179

MoA 0012 SIGNIFICANCE OF CELL ANTIGENS IN SIV VACCINES
Stott James, Mills KHG, Chan WL, Page M, Taffs LF, Kitchin
PA. National Institute for Biological Standards and Control,
Potters Bar, Herts, UK
Objective: To establish the role of cell antigens in protection against SIV.
Methods: Groups of 4 cynomolgus monkeys were vaccinated with inactivated SIV,
virus-infected C8166 cells, uninfected C8166 or RK-13 cells. Vaccinated macaques
were then challenged intravenously with 10 MID50 of SIV grown in either human
cells or simian peripheral blood cells.
Results: Monkeys vaccinated with inactivated SIV, virus-infected cells or
uninfected C8166 cells were protected against challenge with SIV grown in human
cells but notagainst virus grown in simian cells. Vaccination with RK-13 cells did
not protect against SIV.
Conclusions: Cell antigens alone can induce protection. Cell antigens present in
inactivated virus vaccines are sufficient to account for the protection they
induce. The protection observed is influenced by the cells in which the challenge
virus is grown.
Stott, James, NIBSC, Blanche Lane, South Mimms, Potters Bar,
Herts EN6 3QG. Telephone: [phone] FAX: [phone]

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