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 Previous Article | Table of Contents | Next Article 
Blood, Vol. 92 No. 6 (September 15), 1998:
pp. 1941-1949
IL-2 Adenovector-Transduced Autologous Tumor Cells Induce Antitumor
Immune Responses in Patients With Neuroblastoma
By
Laura Bowman,
Michael Grossmann,
Donna Rill,
Michael Brown,
Wan-yun Zhong,
Barbara Alexander,
Thasia Leimig,
Elaine Coustan-Smith,
Dario Campana,
Jesse Jenkins,
Diane Woods,
Geoffrey Kitchingman,
Elio Vanin, and
Malcolm Brenner
From the Departments of Hematology-Oncology, Pathology, and Virology,
St Jude Children's Research Hospital, Memphis, TN; and the Department
of Pediatrics, University of Tennessee, Memphis College of Medicine,
Memphis, TN.
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ABSTRACT |
In many different murine models, the immunogenicity of tumor cells
can be increased by transduction with a range of immunostimulatory genes, inducing an immune response that causes regression of
pre-existing unmodified tumor cells. To investigate the relevance of
these animal models to pediatric malignancy, we used autologous
unirradiated tumor cells transduced with an adenovirus-IL-2 to immunize
10 children with advanced neuroblastoma. In a dose-escalation study, we
found that this tumor immunogen induced a moderate local inflammatory response consisting predominantly of CD4+ T lymphocytes,
and a systemic response, with a rise in circulating CD25+
and DR+ CD3+ T cells. Patients also made a
specific antitumor response, manifest by an IgG antitumor antibody and
increased cytotoxic T-cell killing of autologous tumor cells.
Clinically, five patients had tumor responses after the tumor immunogen
alone (one complete tumor response, one partial response, and three
with stable disease). Four of these five patients were shown to have
coexisting antitumor cytotoxic activity, as opposed to only one of the
patients with nonresponsive disease. These results show a promising
correlation between preclinical observations and clinical outcome in
this disease, and support further exploration of the approach for
malignant diseases of children.
© 1998 by The American Society of Hematology.
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INTRODUCTION |
THE IMMUNOGENICITY of neoplastic cells
can be enhanced by transducing them with genes that encode cytokines or
other immunostimulatory agents. In animal models, these transduced
cells induce an immune response against unmodified tumor cells as well
as against modified cells and may lead to eradication of even
established solid tumors and hematologic malignancies.1-8
Because the cytokines are produced locally by transduced tumor cells,
they do not cause the adverse effects associated with administration of
the high systemic doses of cytokines otherwise needed to induce
antitumor responses. Clinical studies in adults using this approach
have produced variable, but often limited systemic immunologic and
antitumor effects.9,10
Neuroblastoma is the most common extracranial solid tumor of childhood.
The tumor is derived from embryonic neuroectoderm and when it occurs in
infants, it is frequently localized and responds well to
therapy11 or undergoes spontaneous
remission.12,13 In older children, the prognosis is far
worse.14 Although patients with localized disease may still
be cured by conventional therapy, 80% or more of those with
disseminated tumor can be expected to have a relapse within 3 years,
and virtually none of this subgroup will become long-term
survivors.14-17 Over the past decade, attempts to improve
the outcome of advanced neuroblastoma have focused on greater
intensification of the induction and consolidation phases of
chemoradiotherapy, with or without hematopoietic stem cell
rescue.15,16,18 Although remission rates have risen, long-term survival rates have improved only marginally. This failure has prompted a resurgence of interest in alternative methods of disease
eradication,19 particularly immune
modulation.20,21
Several aspects of neuroblastoma tumor biology suggest that the immune
system may be exploited to eradicate the tumor.12,13 Because neuroblastoma is derived from embryonic neuroectoderm, it
expresses antigens not widely detected in the comparatively "mature" tissues of children22-24 and may overexpress
other cellular antigens,14 including some that are
presented on the cell surface.25,26 Moreover, in animals
and in man, neuroblastoma cells are susceptible in vitro and in vivo to
cytotoxic effector mechanisms27-29 induced by cytokines
such as IL-2,30 and to monoclonal antibodies (MoAbs) against neuroectoderm-restricted antigens such as GD2.31-36
We have, therefore, investigated the effects of transferring the IL-2
gene to malignant neuroblasts in children with advanced disease. Based
on data from a murine model,37 we chose to use an
adenoviral vector to introduce the gene into autologous neuroblasts, and to inject escalating doses of these gene-modified cells into patients with relapsed neuroblastoma. We found evidence not only of a
local T-helper cell response associated with destruction of the
injected tumor cells, but also of a systemic antitumor immune response
that involved both helper and cytotoxic T lymphocytes and was
associated with systemic tumor reduction. These results suggest that
IL-2 transduction of neuroblasts provides an immunotherapeutic stimulus, and that adenoviral vectors are useful for this clinical approach.
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MATERIALS AND METHODS |
Patients.
A detailed clinical protocol has been described previously and was
approved by our Institutional Review Board, the Food and Drug
Adminstration (FDA), and the Recombinant DNA Advisory committee of the
National Institutes of Health (NIH).38 Briefly, patients were eligible for study if they had neuroblastoma that had relapsed after one or more intensive courses of multiagent chemotherapy, with or
without autologous stem cell rescue. Although the presence of
measurable disease was not an entry criteria, all patients on study
were potentially evaluable for disease response. Patients were enrolled
not less than 4 weeks after any preceding therapy. Exclusion criteria
included a serum creatinine >3 times normal for age, hepatic
transaminases >3 times normal, Karnofsky score <60%, rapidly
progressive disease, and/or a life expectancy <6 weeks (the
evaluation period of the study). Although the presence of n-myc
amplification was not a direct exclusion criterion, in fact none of the
patients on study had this mutation. Patients were monitored for local
and systemic toxicity by physical examination and blood chemistry
analysis at weekly intervals. Patients received two injections of
autologous neuroblasts 1-week apart, with the second dose 10-fold
higher than the first injection. If both injections were tolerated, and
there was no evidence of rapid progression, patients received two
additional weekly injections at the higher dose. All injections were
given subcutaneously. We used a dose-escalation study, giving between
104/105 and 106/107
gene-modified neuroblastoma cells/kg, up to a maximum of
108 cells/child. Three children were entered at each of the
first two dose levels, and four at the third (n=10).
Tumor response.
At 6 weeks, disease status was determined by clinical assessment,
two-site bone marrow aspiration and biopsy, and by imaging (computed
tomography [CT] or magnetic resonance imaging [MRI] scan of the
chest and abdomen and/or isotope bone scans) where appropriate.
Patients were reassessed at 3-month intervals or more frequently as
clinically indicated. Immunologic assays were performed at 1- to 2-week
intervals for 6 to 8 weeks after the first injection. A complete
response (CR) was defined as complete resolution of all disease
symptoms and signs and regression of all measurable disease. Very good
partial response (VGPR) was defined as >90% reduction in measurable
disease (determined from clinical examination and imaging), whereas
partial response (PR) was defined as >50% but <90% reduction.
Progressive disease was defined as >25% increase in the extent of
established disease, or the appearance of new lesions. These response
criteria have been described in detail elsewhere.38
Adenoviral vector.
The adenoviral vector Ad-IL-2 was constructed by recombining the
plasmid pAVs6-IL2 (linearized with NotI), with the ClaI fragment of
Ad-dl327, an E3 deletion mutant of human adenovirus serotype 5 that has
been used in a cystic fibrosis protocol.39 We have described the structure, production, and activity of the adenoviral vector previously.39,40 Briefly, the vector lacks E1a, E1b, and E3 regions and thus is replication incompetent and devoid of genes
able to inhibit immune responses.
Isolation and transduction of neuroblastoma cells.
Fresh neuroblastoma cells were obtained for transduction from 10 mL to
30 mL of bone marrow or from physically disrupted tumor biopsy
specimens. Cells were initially allowed to adhere to tissue culture
flasks (Collaborative Biomedical, Bedford MA) at a density of
106/cm2 in RPMI supplemented with 10% fetal
calf serum (FCS). After 48 hours, nonadherent cells were discarded, and
trypsinised adherent cells were stained with a panel of five MoAbs
(UJ13A, UJ27.11, M340, Thy1, 5H11.1) that react with >95% of
neuroblastoma cells from >90% of patients. The cells that bound the
antibody were then separated by magnetic beads or by high-speed cell
sorting. This technology allowed us to isolate large numbers of
neuroblastoma cells of >90% purity. Cells obtained in this manner
were transduced at a multiplicity of infection (m.o.i.) of
0.5 to 2 in EHS Matrix T-75 flasks (Collaborative Biomedical) at a cell
density of 105/cm2, to induce IL-2
secretion at levels between 2,000 pg/106 cells/24 hours and
5,000 pg/106 cells/24 hours. After 24 hours, cells were
washed in serum-free medium and cryopreserved. Detailed standard
operating procedures for cell preparation and transduction are
available on request.
Phenotyping of local lesions.
Six to eight µm cryostat sections were prepared from snap-frozen skin
biopsies taken 1 week after each of the first two injections. Sections
were fixed in acetone for 10 minutes at 20°C. After rehydrating in
phosphate-buffered saline (PBS) MoAbs were added. These were anti-CD3
(Dako, Carpinteria, CA), -CD4 (Becton Dickinson, San Jose, CA), -CD8
(RDI, Flanders, NJ), -GD2 (produced in the laboratory from clone MoAb
126 obtained from the American Type Culture Collection, Rockville, MD),
and the Diversi-T panel of anti-T-cell receptor variable regions V
5(a,b and c), V 6.7, V 8(a), V 12, V 2 (T Cell Diagnostics,
Cambridge, MA), or isotype-matched unreactive control antibodies
(Dako). After 30 minutes of incubation, sections were washed in PBS for
10 minutes and incubated for another 30 minutes with goat
antisera to mouse IgG conjugated to fluorescein isothyocyanate (FITC;
Jackson Immunoresearch, West Grove, PA) or to mouse IgM conjugated to
Texas Red (TXR; Southern Biotechnology Associates, Birmingham, AL).
After a final wash in PBS, sections were mounted with a mixture
glycerol:PBS 9:1 containing p-phenylenediamine (1 mg/mL; Sigma, St.
Louis, MO) as antifading agent. Slides were examined with a Zeiss
Axioplan (Zeiss, Thornwood, NY) immunofluorescence microscope equipped with blocking filter for FITC and TXR.
Measurement of antineuroblastoma antibody production.
Nontransduced neuroblastoma cells from each patient (obtained as
described above) were incubated with 100 µL of autologous plasma
obtained immediately before or 3 to 6 weeks after vaccination. Bound
IgG was shown by biotinylated F(ab)2 fragments of donkey antihuman IgG (Jackson ImmunoResearch) then Neutralite-Avidin-R-PE (Southern Biotechnology Associates). All incubation steps comprised 10 minutes at room temperature. The two detection steps were repeated. A
FACScan instrument (Becton Dickinson) was used to analyze
105 cells.
Phenotyping.
Peripheral blood mononuclear cells were phenotyped before and after
immunization by flow cytometry analysis (FACScan, Becton Dickinson)
using the following antibodies: HLA Class I A, B, and C (Olympus, Lake
Success, NY); HLA Class II DR (Becton Dickinson); and CD4, CD8, CD25,
and CD56 (Becton Dickinson). Cells were stained according to the
manufacturers' recommendations, and isotype-matched negative controls
were used for all antibodies.
Cytotoxicity assays.
Two types of assay were used to measure the effects of
AdIL-2-transduced neuroblasts on cytotoxic effector function:
Statistical analysis.
Phenotypic data and cytotoxic activity before and during treatment were
compared by paired t testing.
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RESULTS |
Patients
All patients in this study had relapsed neuroblastoma (patient details
are listed in Table 1). AdIL-2
gene-modified neuroblast doses were escalated as described in Materials
and Methods, so that each child received between 105 and
107 modified neuroblasts/kg per subcutaneous injection, up
to a maximum of 108 cells per dose. Ten patients entered on
the study.
The injections produced minimal adverse effects. A grade 1-2 inflammatory response was consistently observed at the injection site
in patients receiving 105 or more cells/kg, and all but two
of these patients also developed myalgia, which persisted for up to 2 weeks after the last injection. One patient had a more severe,
influenza-like illness (patient 9), but was found to have a rapidly
progressive malignancy to which these symptoms might have been related.
Replication-competent adenovirus was not cultured from any patient, and
no adenovirus vector was detected in peripheral blood samples by
polymerase chain reaction (PCR) analysis.
Local Immune Response
Examination of injection-site biopsies taken after 1 week showed a mild
panniculitis with scanty infiltrating monocytes but a more apparent
infiltration of CD3+ T cells around necrotic tumor cells.
In 7 of the 10 patients the T-cell infiltrate was classified as
moderate (10 to 100 cells/hpf: Fig 1A), in 2 it was light
(<10 cells/hpf) , and in one heavy (>100 cells/hpf). Differential
staining with CD4 and CD8 antibodies showed a preponderance of
CD4+ T cells over CD8+, with a median ratio of
10:1 (range, 2:1 to 100:1) (Fig 1A and 1B). Staining with V -specific
antibodies (see Materials and Methods) showed that the infiltrate was
polyclonal with no evidence of receptor skewing (data not shown).
Finally, staining with the GD2 MoAb showed scanty, mainly necrotic,
residual neuroblasts (Fig 1C, 1D)
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| Fig 1.
Immunofluorescence studies of frozen sections
of tissue taken from the injection sites of patients 3 and 7 one week
after injection of IL-2-transduced autologous tumor cells. The figures
show a moderate infiltration with T cells (A), and scanty surviving
neuroblasts (B). (A,B) Patient 3 (A) and 7 (B) CD4+ T
cells are green, and CD8+ T cells red; there is a
preponderance of CD4+ cells. (C,D) Patient 3 (C) and 7 (D) CD4+ T cells are green, GD2+
neuroblasts are red.
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Systemic Immune Response: Natural Killer (NK) Cells, T-Helper
Lymphocytes, and Cytotoxic T-Lymphocytes
NK cell and T-lymphocyte populations.
Changes observed in systemic immunity involved both NK cells and T
lymphocytes. Phenotypically (Fig 2A and B),
there was a modest but consistent and statistically significant rise in
the proportions and absolute numbers of
CD3+HLADR+ and
CD3+CD25+ cells in the absence of any change in
the overall proportions or numbers of CD3+ T cells. The
mean proportion of CD3+CD4+ cells changed
little during treatment (from 28.2% ± 4.9% standard error
[SE] to 32% ± 5.3%), whereas
CD3+CD8+ cells were similarly unaffected (from
21.4% ± 3.9% to 23.4% ± 4.2%). Figure 2A and B
also shows that there was a small increase in the representation of
CD16+ cells, but CD56 cell percentages and numbers were
unaffected by the treatment. No shift was observed in Vb-receptor usage
(data not shown), indicating the responses induced were likely
polyclonal. There was also a significant rise in the proportions and
absolute numbers of circulating eosinophils and monocytes
(Fig 3A and B).
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| Fig 2.
Phenotypic changes in patient cell populations before
treatment and 0 to 7 days after last injection. (A) Bars represent the
mean proportion ± SEM (n = 10) of cells positive for each marker
combination at each study point. (B) Bars represent the absolute number
(per µL) ± standard error of mean (SEM) (n = 10) of cells
positive for each marker combination at each study point.
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| Fig 3.
Changes in monocytes and eosinophils before treatment and
0 to 7 days after last injection. (A) Bars represent the mean
proportion ± SEM (n = 10) of monocytes and eosinophils. each
study point. (B) Bars represent the absolute number (per microliter) ± SEM (n = 10) of monocytes and eosinophils.
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Tumor-specific responses.
The limited phenotypic changes observed were associated with much more
striking increases in the specific responses to the tumor. In the
T-lymphocyte population, there was recruitment of both helper and
cytotoxic T-cell responses with antitumor specificity. Helper T-cell
activity was evident from the development of IgG antitumor antibodies
postimmunization in four of nine immunologically evaluable patients
(Fig 4).
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| Fig 4.
(A-D) Appearance of IgG antibody to tumor cells after
immunization. Pretreatment fluorescence in all patients was at the
level of controls (normal human serum). Three to 6 weeks after the
first immunization, four of the patients analyzed (numbers 1, 3, 5, 6)
who are shown here had increased antibody binding to autologous tumor
cells, shown by an increase in fluorescence activity. Increased
activity persisted at least 3 months after completion of therapy. (E)
For comparison, a nonresponding patient is shown.
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We also investigated whether the adenovirus vector was able to generate
an antiadenovirus response that might affect the efficacy or toxicity
of the vaccine. We found no change in titers of antibody directed to
adenovirus (Table 2), even in patients who
were already highly immune to adenovirus.
Longitudinal analysis of tumor-specific cytotoxicity.
Availability of samples allowed us to make a longitudinal analysis of
tumor-specific cytotoxic activity in eight patients. Initially, PBM
were cultured with nontransduced neuroblasts in a standard 4-hour
Cr51 release assay, to measure circulating tumor-specific
CTL. Figure 5 shows that four patients
(numbers 4, 6, 7, and 8) showed a significant and substantial rise in
CTL activity against autologous targets. In patient 6, marked elevation
was observed only at one time point. However, this cytotoxicity was
titratable through all E:T ratios tested (from 51% to 11%). In all
patients with measurable cytotoxicity, addition of anti-Class-I MHC
antibody reduced killing by a mean of 57% (range, 23% to 91%;
P< .001), suggesting that a significant proportion of the
cells responsible for effector function were classical CTLs. Of the
patients whose freshly isolated PBM showed no increased killing of
tumor cells, one (patient 2) showed a 40-fold rise in
neuroblastoma-specific CTL precursor frequency during treatment
(Fig 6). Hence, five of the eight patients
examined for a cytotoxic antitumor response showed a significant
increase in activity during treatment. These shifts in tumor-specific
response were not associated with any consistent alteration in NK
activity (measured by killing of K562 cells), which remained
essentially unchanged throughout the treatment period
(Fig 7).
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| Fig 5.
Longitudinal analysis of cytotoxic activity against
autologous neuroblasts. Patient lymphocytes were cultured with
51Cr-labeled autologous neuroblasts at an E:T ratio of
40:1, 20:1, and 10:1 at increasing time intervals after immunization.
Each line shows specific isotope release from a single patient, at an
E:T ratio of 40:1.
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| Fig 6.
Rise in CTL precursor frequency. Patients who showed no
increase in direct cytotoxic activity were examined for a rise in CTL
precursor frequency of these individually. Patient 2 (shown here)
showed a 40-fold rise over the treatment period.
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| Fig 7.
Longitudinal analysis of natural killer cell activity.
Patient lymphocytes were cultured with 51Cr-labeled K562
cells at effector:target ratios of 20:1,10:1, and 5:1 at increasing
time intervals after immunization. Each line shows specific isotope
release from a single patient at an E:T ratio of 20:1.
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Tumor Response
Results are summarized in Table 3. At the
initial 6-week evaluation, patient 7 showed regression of her
paratracheal mass. Four additional patients (numbers 2, 4, 8, and 10)
were initially classified as having stable disease at 6 weeks. Of these
individuals, patient 8 with extensive marrow involvement subsequently
entered complete remission and remains free of measurable disease >6
months after therapy, whereas patient 10 remains with stable disease. Patients 2 and 4 were treated with oral etoposide immediately after the
evaluation period: patient 2 had a >90% tumor response, and patient
4 entered a complete remission that has been sustained for >319 days.
Overall, 3 of 10 patients showed tumor responses (1 complete, 1 partial, 1 stable disease) to the vaccine alone, whereas 3 additional
patients responded (1 complete, 2 partial) to therapy with a
combination of vaccine followed by oral etoposide.
Of the 5 patients who had stable disease or tumor regression at week 6 (patients 2, 4, 7, 8, and 10), 4 were evaluated for antitumor cytotoxic
responses; all had coexisting activity (Figs 5 and 6);
conversely, of the 4 patients with progressive disease, only 1 (patient 6) made an antitumor cytotoxic response. Of the 10 patients
studied, 7 are alive 150 to 485 days after completing treatment; 6 have
measurable residual disease.
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DISCUSSION |
Studies in various animal models have shown that tumor cells, including
IL-2-transduced neuroblasts, can generate a systemic immune response
capable of delaying the growth of unmodified tumor cells that express
the same tumor-related or tumor-specific antigens.1-8 As we
have shown, these observations can translate well into clinical practice. Both a local and a systemic antitumor immune response were
generated in children given injections of autologous unirradiated neuroblastoma cells gene-modified to secrete IL-2. Although the local
response consisted predominantly of CD4 T cells, the systemic antitumor
response was more broadly based, and included both a T-helper cell
response (manifested by antitumor antibody production) and increased
cytotoxic T- lymphocyte activity directed against autologous tumor
cells. CTL activity was discernible both by direct killing of
autologous neuroblasts and by an increase in the specific CTL-precursor
frequency. There was also evidence for limited recruitment of
nonspecific effector mechanisms, with an increase in monocytes and
eosinophils.
Although a potent increase in antitumor activity was observed
after injection of IL-2-modified neuroblasts, there was no
discernible increase in humoral antiadenoviral immune response, even in
patients with prior adenovirus immunity. Presumably, the ex vivo
transduction protocol did not transfer sufficient virus particles to
induce a virion-specific response, whereas the E1/E3-deleted vector
itself likely expresses too few adenovirus antigens in neuroblastoma cells to induce an adenovirus-specific antibody response through the
endogenous antigen processing pathway.42,43 We were unable to obtain sufficient material from these pediatric patients to discover
whether a cellular immune response to the adenovirus vector developed.
If such a response does occur, it evidently does not destroy the
infected cells so rapidly as to preclude a substantial antitumor
response.42,43
Injection of the AdIL-2-transduced tumor cells was well tolerated apart
from systemic myalgia. Because we did not detect an antiadenovector
immune response, and because no patient released measurable adenovirus
after treatment, we believe the systemic symptoms were a consequence of
the immune activation induced by the gene-modified cells. Certainly,
systemic recruitment of T cells and eosinophils requires release of a
range of cytokines, including IL-1, TNF, IL-6, gamma interferon, and
IL-2, any of which may have contributed to the patients'
symptoms. We are now investigating whether systemic cytokine release
could account for the adverse effects.44
At the initial 6-week evaluation, four patients had stable disease and
only one had shown a PR. Subsequently, however, one of the patients
with stable disease progressed to a complete tumor response. Hence, in
contrast to the relatively rapid responses usually observed after
chemotherapy and radiotherapy, a clinically evident antitumor immune
response may require many weeks, and evaluation should be
correspondingly delayed. Our data also suggest a link between the
development of antitumor immunity and the detection of a tumor
response. Four of five patients with tumor regression or stable disease
had a tumor-specific CTL response, whereas only one of the four
patients with progressive disease had antitumor activity. In this last
individual (patient 6), a lymph node taken from a site of disease
progression was densely infiltrated with T cells, with only scanty
residual tumor. Although these data are consistent with a link between
CTL activity and tumor response, this connection will need to be
confirmed in a larger study. It is less likely that modulation of NK
cells contributed to the antitumor response. NK activity showed little
or no change in response to immuniztion, and primary neuroblasts have
only limited susceptibility to killing by this effector
population.27,28 If a link between CTL activity and disease
response is genuine, subsequent disease persistence or progression may
be caused by either the gradual dwindling of the antitumor response or
the emergence of antigenic loss variants of neuroblastoma. This
distinction is an important one. If late relapse is due to a decline in
immune responsiveness, then repeated immunization over a prolonged
period may be required. If recurrence is due to the emergence of
antigenic loss variants, the approach might best be limited to
individuals with minimal residual disease in whom the risk of further
tumor mutation would be lower.
We have shown that IL-2-transduced autologous neuroblasts can induce
antitumor immune responses in patients with advanced neuroblastoma. How
may the benefits of this approach be increased? One possibility is to
transfer alternative immunostimulatory genes. Human tumor cells have
been transduced with genes encoding many such molecules, and of these,
granulocyte-macrophage colony-stimulating factor (GM-CSF) and HLA-B7
have been reported to induce localized antitumor
immunity.9,10,45 We cannot directly compare the immunologic
or antitumor potency of these alternative agents with our own approach,
because none of the other studies were performed in children with
neuroblastoma. Although other single immunostimulatory molecules may
possibly produce an antineuroblastoma response superior to that of
IL-2, we believe that to obtain the optimal T-cell immune response to
human tumors from this approach will require combining different
classes of immunostimulatory molecules. In animal models, for example,
the combination of the T-lymphocyte chemokine lymphotactin and the T
lymphocyte growth factor IL-2 enhance and accelerate antitumor
responses as compared with any one agent alone.46,47
Because current single-agent genetic immunotherapy approaches evidently
produce effective systemic antitumor responses in humans, there is
reason to hope that these molecular combinations will be successful in
the clinical setting. To be effective, the combinatorial approach will
require the availability of safe vectors that can rapidly and reliably
transduce a high percentage of primary human tumor cells. For
neuroblastoma at least, our results show that adenovirus vectors meet
these requirements.
In conclusion, we have evidence that clinically effective antitumor
immune responses can be safely produced in children with advanced
neuroblastoma by adenovector-mediated transfer of the IL-2 gene. It
will now be of interest to discover whether the immune response and
antitumor response can be further enhanced by appropriate combinations
of immunostimulatory molecules.48
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FOOTNOTES |
Submitted March 5, 1998;
accepted May 8, 1998.
Supported by National Institutes of Health Core Grants No. RO1 CA58211
and P30 CA21765, by the Assisi foundation, and by ALSAC (American, Lebanese, Syrian Associated Charities).
Address correspondence to Malcolm K. Brenner, Texas Children's
Hospital, 6621 Fannin MC 3-3320, Houston, TX 77030.
The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" is accordance with 18 U.S.C. section 1734 solely to indicate this fact.
| |
ACKNOWLEDGMENT |
We would like to thank Jing Feng Zhao, Marti Holladay, and Michael
Khouri for technical assistance; Sharon Naron for scientific editing;
and Tracie Keahey for word processing. We are grateful to all our
medical colleagues who referred patients for this study.
| |
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Abstract
Introduction
Abstract