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Clinical Trials |
The Wistar Institute [S. V., A. C., D. S.], The University of Pennsylvania Cancer Center [D. L. P., S. L. L., L. S., M. K., K. D., E. A. S.], and The Fox Chase Cancer Center [M. H. T.], Philadelphia, Pennsylvania 19104
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ABSTRACT |
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 Top ABSTRACT INTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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,
interleukin-10, and activation markers (soluble interleukin-2 receptor
and soluble intercellular adhesion molecule-1) were often seen.
Only one patient developed anti-HLA class I antibody responses against
TALL-104 cells; specific CTL activity developed in three patients
during induction and in four patients during the maintenance boosts. In
conclusion, TALL-104 cells were well tolerated by patients with
metastatic breast cancer at the doses and regimen tested. The clinical
responses observed in this preliminary trial demonstrate that further
investigation of TALL-104 cell therapy is warranted. |
INTRODUCTION |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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The human major histocompatibility complex nonrestricted killer cell line TALL-104 (CD3+, CD8+, CD56+, CD16-) developed in this laboratory (2 , 3) might represent a new immunotherapeutic approach to cancer, as suggested by evidence in animal models. Specifically, adoptive transfer of lethally irradiated (40 Gy) TALL-104 cells into SCID3 mice has induced regression of transplantable human hematopoietic and nonhematopoietic malignancies (4, 5, 6, 7, 8) . In addition, remarkable antitumor effects were seen in immunocompetent mice bearing syngeneic leukemia (9) and in pet dogs with spontaneous tumors (10, 11, 12) . Although TALL-104 cell killing in vitro is mostly mediated by necrotic mechanisms involving the release of cytotoxic factors (granzymes, perforin, and TIA-1; Ref. 3 ), several tumor targets die by apoptosis (7 , 9) ; in addition, cytostatic mediators and cytokines (e.g., TNF), released by TALL-104 cells upon interaction with the tumors (5 , 10, 11, 12) , play a role in the induction of tumor death. In vivo, TALL-104 cells have been shown to induce tumor necrosis (5 , 10) ; moreover, multiple systemic delivery of TALL-104 cells into immunocompetent animals was followed by the development of antitumor immunity (immunological memory) that protected the animals from tumor rechallenge (in mice) or progression (in dogs; Refs. 9, 10, 11 ). Biodistribution studies in healthy animals showed that 111In-labeled TALL-104 cells, injected i.v, rapidly localized in the lungs, liver, and kidneys; 2 h after injection, the lungs were cleared, whereas liver, kidney, spleen, and bone marrow were the organs with major TALL-104 cell accumulation in the following 24 h (13 , 14) . Importantly, in tumor-bearing animals, 111In-labeled TALL-104 cells were present both within the primary tumor mass and at the site of distant metastases (6 , 14) .
Our previous preclinical studies in SCID mice engrafted with human breast carcinomas (6 , 14) and in pet dogs and cats with spontaneous metastatic mammary tumors (15) 4 have shown that breast cancer is highly sensitive to the antitumor effects of TALL-104 cells. These encouraging results prompted us to initiate this Phase I clinical trial to explore the safety and potential efficacy of escalating doses of lethally irradiated TALL-104 cells administered systemically to women with refractory metastatic breast cancer. The maximal dose tested (108/kg/day) was the one proven effective in preclinical veterinary studies (10, 11, 12 , 15) .
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MATERIALS AND METHODS |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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. Other eligibility criteria
included age >18 years, performance status 0–1 (by the criteria of
the Eastern Cooperative Oncology Group), life expectancy >8 weeks, and
adequate hematopoietic (absolute neutrophil count
>1,500/ml3 and platelets
>100,000/ml3), hepatic (total bilirubin level
<1.5 mg/dl), and renal (creatinine concentration <1.5 mg/dl)
functions.
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Trial Design.
The trial was designed as a single-center, open-label, dose-escalation
study. The schedule of administration included a first 5-day cycle
(induction course) of lethally irradiated (40 Gy) TALL-104 cells
administered by i.v. infusion (100 ml) over 60 min, followed by 2-day
monthly boosts until disease progression or limiting toxicity (Fig. 1)
. Five dose levels of TALL-104 cells
(106/kg, 3 x 106/kg,
107/kg, 3 x 107/kg,
and 108/kg) were tested (three patients/dose
level; Table 2
). No intrapatient dose
escalation was allowed. Twenty-eight treatment courses were
administered in this trial, for a total of 101 cell injections (Table 2)
. All patients received premedication with diphenylhydramine and
acetaminophen within 1 h before the start of each cell infusion.
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Manufacturing of the TALL-104 Clinical Product.
Manufacturing of clinical grade TALL-104 cells was performed under Good
Laboratory Practice conditions in a specially designated facility at
The Wistar Institute. TALL-104 cells were grown in endotoxin-free
Iscove’s modified Dulbecco’s medium (Life Technologies, Inc., Grand
Island, NY) supplemented with 10% heat-inactivated fetal bovine serum
(Atlanta Biologicals, Norcross, GA) and 100 units/ml recombinant human
IL-2 (Chiron Therapeutics, Emeryville, CA) in humidified incubators at
37°C with 10% CO2 in T-175 vented-cap flasks
(Falcon, Franklin Lakes, NJ). Mycoplasma contamination was
monitored weekly on cell samples taken from at least two flasks from
each incubator using a commercial PCR kit (American Type Culture
Collection, Rockville, MD). Three times a week, cells were harvested by
centrifugation in 250-ml conical tubes (Corning, New York, NY), washed
twice in saline (Abbott Laboratories, King of Prussia, PA),
-irradiated (40 Gy) using a 137Cs source,
resuspended in freezing medium consisting of human plasma protein
fraction 5% (American Red Cross Blood Services, Philadelphia, PA) and
10% DMSO (Rimso; Tera Pharmaceuticals, Buena Park, CA), and then
transferred into Fenwal transfer pack containers (Baxter Healthcare
Corporation, Deerfield, IL). The TALL-104 cell packs were stored at
-70°C. Each lot of frozen TALL-104 cell packs was tested for
sterility, Mycoplasma contamination, endotoxin, purity,
identity, phenotype, proliferation, and cytotoxic function (quality
control assays; Table 3
). Only lots that
met the standard criteria summarized in Table 3
were used for patient
administration. When needed for infusion, frozen TALL-104 cell packs
were rapidly thawed in a 37°C water bath, and saline was added to
reach a final volume of 100 ml. Cell aliquots were removed from each
pack for determination of cytotoxic and proliferative activities and
sterility (quality control assays). The TALL-104 cell packs were then
handed to the clinical personnel for patient injection, as described
above. A Gram’s stain (statim) was performed at Hospital of the
University of Pennsylvania just before infusion.
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When available, tumor cells harvested from patients’ tumor biopsies by mechanical dissociation (6 , 17) were cultured briefly and tested in 18-h 51Cr release assays for susceptibility to the lytic activity of TALL-104 cells and autologous PBMCs.
Monitoring of anti-TALL-104 cell antibodies in the patients’ sera was performed by immunofluorescence. Briefly, the sera were diluted 1:100 and 1:1000 in FACS buffer (9, 10, 11, 12 , 15) and incubated for 1 h with the TALL-104 cells. After three washes in FACS buffer, FITC-conjugated goat antihuman IgG (whole molecule) and antihuman IgM (µ-chain specific; Sigma Chemical Co., St. Louis, MO) were added at a 1:100 dilution in FACS buffer for another hour. After final washes in FACS buffer, the percentage of reactivity of the sera with TALL-104 cells was calculated as described previously (9, 10, 11, 12 , 15) . HLA class I antibody screening in the patients’ sera was performed using a cell panel from 35 volunteers and the AHG-CDC assay. Furthermore, indirect immunofluorescence flow cytometry technique using HLA class I microbeads (One Lamba, Inc., Canoga Park, CA) was performed to specifically detect anti-HLA class I antibodies, as described (18 , 19) .
Variations in serum levels of two markers for nonspecific immune activation, i.e., sIL2-R and sICAM-1, were also investigated, using commercially available ELISA kits (Endogen, Woburn, MA), according to the manufacturer’s instructions. The sensitivity of the assays was 24 units/ml for sIL2-R and 0.3 ng/ml for sICAM-1.
Cytokine Assays.
Patients’ sera were collected pre-TALL-104 cell infusion (time 0) and
at 2, 4, and 6 h after TALL-104 cell infusion; on day 1, before
each TALL-104 infusion on days 2, 3, and 5; and weekly thereafter until
disease progression. The presence of IFN-, TNF-
, TNF-ß, GM-CSF,
and IL-10 in the sera was tested using human cytokine-specific ELISA
kits, according to the manufacturer’s instructions. The sensitivity of
the assays was 2 pg/ml for IFN- and GM-CSF, 5 pg/ml for TNF-, 3
pg/ml for IL-10 (Endogen), and 7 pg/ml for TNF-ß (R&D, Minneapolis,
MN).
PCR Analysis.
Patient PBMC samples, obtained immediately before each TALL-104 cell
infusion, daily during infusions, and at subsequent visits after cell
infusions, were subjected to DNA extraction using standard techniques
(6
, 10, 11, 12
, 15)
. The presence of circulating TALL-104
cells in each cell extract was evaluated by PCR analysis using two
primers specific for the human minisatellite region YNZ.22. An
oligonucleotide probe recognizing 24 nucleotides in the middle of the
amplified sequence was used to demonstrate the specificity of the PCR
products by Southern blot hybridization, as described previously
(6
, 10, 11, 12
, 15)
.
Statistical Analysis.
The significance of hematological and immunological changes within each
patient during and after TALL-104 cell infusions was tested by paired
Student’s t test.
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RESULTS |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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. Median age was 49 years (range, 33–63 years);
the primary diagnosis was infiltrating ductal carcinoma (12 patients),
infiltrating lobular carcinoma (2 patients), and medullary carcinoma (1
patient). All but one patient had failed a combination of at least two
different treatments including chemotherapy, radiation therapy,
hormonal therapy, immunotherapy with HER2/neu antibodies, or high-dose
chemotherapy with SCT. One patient (008) had progressed after only one
chemotherapy regimen but was ineligible for further chemotherapy
because of a severe cardiopulmonary syndrome. Two patients (001 and
003) had platelets <100,000/ml (3)
attributable to a
history of autologous SCT, two patients (010 and 011) had increased
alkaline phosphatase levels attributable to metastatic disease, and one
patient (015) had a history of brain metastases.
Clinical Responses.
Nine patients had progressive disease by 1 month after the induction
course and received no further therapy with TALL-104 cells. Five
patients receiving 106 (n = 2),
3 x 106 (n = 2), and
108 (n = 1) cells/kg had SD for
2–6 months after the induction course, thus qualifying for monthly
maintenance boosts; among them, patient 001 had a significant reduction
in narcotic requirements for bone pain. One patient (011) had a
decrease in size of liver metastases (many of the lesions having become
necrotic) and in the amount of perihepatic and pelvic fluid 3 weeks
after receiving the TALL-104 induction course at 3 x
107/kg. Unfortunately, this patient withdrew from
the study for personal reasons before receiving the first monthly
boost, precluding further evaluation. As of February 2000 (16 months
after the end of the study), three patients (013, 014, and 015) are
still alive with active disease. Elevated levels of the CEA tumor
marker (492, 68.6, 254, and 175.5 units/l) were present pretreatment in
four patients (001, 009, 011, and 014, respectively). Only 1 of these
patients (001) had SD after the induction course, although the CEA rose
to 2.6 times the pretreatment level, 2 months after the first TALL-104
infusion. No patient had a decrease in CEA levels during this trial.
Toxicity.
No significant toxicity was associated with TALL-104 cell
infusions, although the DMSO present in the cell product was
responsible for taste change and alithosis in almost all patients. One
patient (014) experienced grade IV nausea, vomiting, and fevers related
to necrosis of hepatic metastases, 3 weeks after TALL-104 induction,
but no other grade III and IV toxicities were seen (Table 4)
. Several patients experienced grade
I-II toxicities as outlined in Table 4
, which were transient and of
uncertain relationship to TALL-104 cell infusions. No toxic effects
were observed in any of the patients who underwent maintenance therapy
(not shown).
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Detection of Circulating TALL-104 Cells.
Short-term (1 week) and long-term (up to 2 months) monitoring of
circulating TALL-104 cells was performed in 5 patients by PCR analysis.
TALL-104 cells were detectable in the peripheral blood of all five
patients during the 5-day induction treatment but could no longer be
detected by day 7 from the last cell injection in each cycle (not
shown).
Humoral and Cellular Immune Responses against TALL-104 Cells.
Only one patient (012) developed antibodies against TALL-104 cells;
serum from this patient became 89.1% reactive with TALL-104 cells as
early as day 7 from the start of therapy and remained positive until
the date off study (2 weeks later). By the AHG-CDC bioassay and
indirect immunofluorescence flow cytometry analyses with HLA class I
microbeads, it could be determined that the postimmune serum from
patient 012 reacted with HLA class I antigens, whereas the preimmune
serum was not reactive (Fig. 2)
.
Furthermore, the postimmune serum was reactive by AHG-CDC with the cell
panel. The HLA specificity of this serum was broad and included the
TALL-104 HLA class I antigens (not shown). Importantly, the postimmune
serum did not inhibit TALL-104 killer activity against the patient’s
own tumor biopsy in in vitro cytotoxicity assays (not
shown). No other patient developed detectable antibodies against
TALL-104 cells, including the five patients who received monthly boosts
for 2–6 months (not shown).
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, although in three
patients (002, 007, and 012) specific killing of TALL-104 cells either
developed or increased (Fig. 3B)
, regardless of cell dose.
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. The limited number of patients in the
study and the large variability of NK activity among the patient
population precluded an analysis of the statistical significance of
this observation.
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Four of the tumor specimens were sensitive to in vitro lysis
by TALL-104 cells (Table 5)
. Cytokine
production was detected in the supernatants of all five tumor/TALL-104
cocultures (Table 5
and Fig. 5
). The
levels of cytokines in the cocultures with tumor cells isolated from
lymph node biopsies were low (003, 012, and 015; Fig. 5A)
or
intermediate (009; Fig. 5B)
, whereas those with tumor cells
from the chest wall of one patient (008) were remarkably high (Fig. 5C)
.
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Serum Cytokines and Immune Activation Markers.
The presence of cytokines (IL-10, IFN-, TNF-, TNF-ß, and
GM-CSF) known to be released by TALL-104 cells in vitro
(20)
and in animal studies (10, 11, 12)
upon
interaction with tumor cells was measured in patients’ sera before and
at different time points during the TALL-104 induction course. None of
the patients had detectable serum levels of TNF-, TNF-ß (<5
pg/ml), or GM-CSF (<2 pg/ml) before and during cell treatment, except
for patient 001, who had increased TNF- levels (7.6 pg/ml) starting
on day 4 of the induction course.
IFN- was present in the sera of 13 patients pretreatment and
increased by >20% in three cases, at 2 h (n =
1), 4 h (n = 1), or 5 days (n = 1)
after the first TALL-104 infusion. Two additional patients who had no
detectable levels of IFN- before treatment had significant levels of
this cytokine during the induction course, starting at 4 h on day
1 (patient 008) or on day 3 (patient 005). Nine patients had baseline
elevated levels of IL-10, which increased by >20% in three patients
during treatment, starting at 2 h (014) or 4 h (012) of day 1
or on day 4 (007) of the induction course, independent of TALL-104 cell
dose. Four patients with undetectable levels of IL-10 before treatment
had significant levels of this cytokine starting at 2 h
(n = 1) and 4 h (n = 3) on day 1
of the induction course.
Serum levels of the sIL-2R and sICAM-1 markers, associated with
nonspecific immune activation, were measured at the same time points.
As shown in Fig. 6
, there was a trend
toward an increase in both markers during TALL-104 cell infusions,
which was evident as soon as day 1 after treatment and returned to
baseline by day 5. No statistically significant difference was observed
when high (Fig. 6, A and C)
versus low
(Fig. 6, B and D)
pretreatment levels were
compared.
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) were immunologically
monitored during the 2-day monthly maintenance infusions (see treatment
schema, Fig. 1
). None of these patients had detectable serum levels of
TNF-, TNF-ß, or GM-CSF before or during the boosts. The IFN-
and IL-10 levels observed on day 5 of the induction course consistently
returned to baseline levels before each subsequent monthly boost and
always increased on day 2 of each boost (not shown). In the case of
patient 001, the increased levels of IL-10 (61.8 pg/ml) that developed
4 h after the first TALL-104 infusion were still present in the
patient’s serum up to 3 months after the start of therapy (not shown).
sIL2-R and sICAM-1 serum activation markers followed the same
pattern described for the cytokines, with return to baseline
levels before each TALL-104 cell infusion, followed consistently by
an increase on day 2 of each monthly boost (not shown).
Peripheral blood NK activity against K562 cells was highly variable
among patients and also varied within each individual patient when
tested at different times. An example is shown in Fig. 7
for patient 005, whose NK activity,
seen at day 5 of the induction course, remained stable until after the
second boost; her NK activity increased at this time, stabilized until
administration of the third boost, decreased until administration of
boost 4, and rose again after boost 4 and during boost 5 (6 months
after initiation of cell therapy), reaching the maximum value (Fig. 7)
.
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DISCUSSION |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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Unlike HLA-matched, sibling/donor-derived effector cells, TALL-104 cells represent a universal donor system and provide an unlimited and reliable source of tumoricidal cells with stable cytotoxic activity, which is ideal for adoptive immunotherapy approaches. Although dependent on IL-2 for expression of cytotoxicity and long-term survival in vitro, TALL-104 cells display antitumor effects in animal models when used as a single agent (4, 5, 6, 7, 8, 9, 10, 11, 12 , 15) , thus offering a potentially nontoxic cell therapy approach as compared with LAK or tumor-infiltrating lymphocyte therapies in association with IL-2 (21 , 22) . The major objective of the present study was to evaluate the potential toxicities associated with multiple systemic administrations of lethally irradiated, nondividing, TALL-104 cells in women with metastatic refractory breast cancer. In addition, immunological studies were performed in an attempt to identify surrogate markers for toxicity and/or clinical responses.
Adverse effects from TALL-104 cell therapy were generally minimal, limited to grade I-II toxicity, and were often of uncertain relationship to cell infusions. Mild gastrointestinal toxicity (nausea and vomiting) was observed during TALL-104 treatment in 40% of the treated patients. This has also been reported to occur in 80% of cancer patients during LAK/IL-2 therapy (31) and was reported in 5% of TALL-104-treated dogs with spontaneous tumors in our previous study (10) . Taste change and alithosis was associated with the presence of DMSO used as cryostabilizer in the TALL-104 cell packs. The range of DMSO given with each TALL-104 cell infusion varied from 0.06 to 0.2 g/kg; this dose is much lower than that (0.2–1.3 g/kg) administered during autologous SCT using cryopreserved grafts, perhaps explaining, at least in part, the lack of toxic signs (flushing and pulmonary and abdominal toxicity) attributed to DMSO in those trials (32 , 33) . Another explanation for the low incidence of side effects in our study may be the low level of cell-lysed products infused (TALL-104 cell viability after thawing was 85–90%). Comparatively, in thawed marrow grafts, the final amount of cell lysis products injected may be very high, depending on the quantity of contaminant mature cells present, which are usually poorly preserved (32 , 33) .
No severe toxicities, such as hypotension, secondary to increased capillary permeability, were observed in our study. These effects have been reported to be induced by autologous LAK/IL-2 therapy (34) . Although the mechanism by which IL-2 and LAK cells induce a vascular leak syndrome is unknown, LAK cells have been shown to bind and lyse normal human vascular endothelial cells in vitro (35 , 36) . Whether this applies also to TALL-104 cells is now under investigation in this laboratory. However, the absence of vascular leak syndrome in mice, dogs, cats, and monkeys treated with up to 5 x 108/kg TALL-104 cells in the absence of exogenous sIL-2 (4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 , 37) suggests that this effect does not reach biological significance with TALL-104 cells.
None of the common hematological abnormalities induced by LAK/IL-2 therapy (anemia, lymphopenia followed by rebound lymphocytosis, and thrombocytopenia with coagulation disorders; Ref. 38 ) were detected in the present study. Grade I-II leukopenia with neutropenia was seen in two patients and rapidly reversed once therapy was halted. Most patients developed a mild degree of monocytosis and eosinophilia; this abnormality has also been reported in patients treated with IL-2 and LAK cells (38) .
Previous studies in pet dogs with spontaneous cancers (10, 11, 12
, 15)
indicated that the presence of human (TALL-104 released)
TNF-, TNF-ß, and IFN- in the sera of the treated dogs
correlated with clinical efficacy. In the present studies, no changes
were observed in serum levels of TNF and GM-CSF, but an increase in the
levels of IFN- and IL-10 was observed in the sera of 69 and 27% of
the patients treated, respectively. Although the cytokine profile
observed in the small patient population in this study is difficult to
interpret, such cytokine responses may represent a secondary reaction
against the introduction of the allogeneic TALL-104 cells rather than
being directly involved in an antitumor effect by the cell line. The
cellular source (whether TALL-104 or the host’s immune cells) of
IFN- and IL-10 remains unclear; however, IL-10 has been detected in
primary breast cancers and is associated with the induction of T-cell
anergy (39)
, as a result of T-cell proliferation and
function, reduced IL-2 production, and antigen presentation
(40)
. In a previous study in SCID mice engrafted with a
variety of human solid tumors (8)
, the serum levels of
sICAM-1 in the TALL-104-treated animals correlated with tumor burden
and were indicative of treatment efficacy. In the present study, we
evaluated the changes in serum levels of sICAM-1 and sIL-2R because
these markers are known to be associated with human breast cancer
progression (41
, 42)
. The changes observed, however, did
not reach statistical significance and are difficult to interpret
because of the limited number of patients treated at each dose level.
Although it has been suggested that IL-2-activated lymphocytes are hepatotoxic (38 , 43) , we noted mild and transient liver function abnormalities in only two patients. Severe toxicity (grade IV) was observed in only one patient receiving the highest cell dose and was temporarily related to hepatic tumor necrosis developing 3 weeks after the induction course. Although it is difficult to prove that this toxicity was consequent to TALL-104 cell therapy, it is tempting to speculate that this event reflected a marginal response, based on the fact that TALL-104 cells do induce necrotic tumor cell death in vitro (3 , 16) and in animal studies (4, 5, 6, 7, 8, 9, 10, 11, 12 , 15) . However, no liver tissue was available from this patient to conduct histological and/or in situ hybridization studies to definitively document an antitumor response.
Xenogeneic antibody responses were consistently observed in our preclinical studies involving TALL-104-treated mice, dogs, and monkeys; although such antibodies did not have neutralizing activity in vitro, they were responsible for the progressively faster kinetics of TALL-104 cell clearance with boosts (10, 11, 12, 13 , 15 , 37) . Surprisingly, in the present study, nonneutralizing anti-HLA class I antibodies against TALL-104 cells developed in only 1 of 15 patients. It is possible that the dose, route, and time of TALL-104 cell administration, combined with the host compromised immune status, resulted in a humoral tolerant response versus the allogeneic cells. This explanation would be consistent with previous findings demonstrating that female, first-transplant candidates induced panel-reactive antibodies in only 5–10% of the recipients (44 , 45) . The mechanism of low anti-HLA antibody responses is not known, but it has been suggested that each transfusion induces some degree of clonal expansion that is too low in magnitude to produce a persistent antibody response; with higher numbers of transfusions, an increasing proportion of patients develops a critical mass of memory T and B cells capable of sustaining a persistent antibody response (significant clonal expansion; Refs. 44 and 45 ). In this respect, TALL-104 cells might represent a setting of alloimmunization similar to the one described for allogeneic transfusions and/or transplantations (44 , 45) . A larger number of patients receiving longer periods of treatment needs to be studied to clarify this important issue. Contrary to the antibody responses, specific CTL responses to TALL-104 cells occurred in three patients during the induction course, and in four of five patients over the monthly boosts. Although the cellular responses observed in this immunosuppressed patient population were low, the development of CTL activity to TALL-104 cells might have important therapeutic implications for the design of long-term administration schedules. In fact, our data in healthy dogs (13) clearly indicated a progressively faster clearance of TALL-104 cells from the blood and organs after multiple daily injections as well as at the time of each monthly boost, when host immune responses against the xenogeneic cells had developed. Although the overall lower response to TALL-104 cells observed in cancer patients might not lead to antibody- and cell-mediated rejection of the allogeneic effectors in human trials, studies to analyze the kinetics of distribution and clearance of TALL-104 after multiple injections will be necessary to design effective treatment regimens. One question that needs to be answered is the need for repeated boosts to achieve efficacy; in fact, in preclinical studies (10, 11, 12 , 15) , the number of TALL-104 cell infusions did not seem to correlate with clinical efficacy. Ongoing trials will elucidate whether the administration of higher doses of TALL-104 cells, in a short time period (days) before immune responses develop, would be effective enough to control or eradicate cancer growth through direct tumoricidal effects and/or induction of specific antitumor immunity, thus eliminating the need for long-term boosting.
On the basis of the advanced clinical stage and heavy pretreatment of the 15 patients in this Phase I study, observing even a marginal clinical response in one patient and noting significant pain relief in a second patient is encouraging. These findings, together with the disease stabilization seen in five patients for 2–6 months, suggest biological activity of TALL-104 cells and point to their potential clinical benefit. Clinical trials have been planned to identify patient populations with highly responsive tumor types.
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ACKNOWLEDGMENTS |
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FOOTNOTES |
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1 Supported by United States Army Medical Research
and Materiel Command and partially by the Breast Cancer Fund,
the Expedition Inspiration Fund, a gift from Felicity and Peter
Benoliel, and the NIH Core Grant CA10815-30. 
2 To whom requests for reprints should be
addressed, at The Wistar Institute, 3601 Spruce Street, Philadelphia,
PA 19104. Phone: (215) 898-3978; Fax: (215) 573-7919; E-mail: santoli{at}wistar.upenn.edu
3 The abbreviations used are: SCID, severe
combined immunodeficiency; TNF, tumor necrosis factor; IL, interleukin;
sIL-2R, soluble IL-2 receptor; sICAM, soluble intercellular adhesion
molecule; SCT, stem cell transplantation; CEA, carcinoembryonic
antigen; PBMC, peripheral blood mononuclear cell; NK, natural
killer; FACS, fluorescence-activated cell sorting; HLA,
histocompatibility antigen; AHG-CDC, antihuman globulin-enhanced
complement-dependent lymphocytotoxicity; GM-CSF,
granulocyte/macrophage-colony stimulating factor; SD, stable disease;
LAK, lymphokine-activated killer.
4 S. Visonneau, K. A. Jeglum, and D. Santoli,
unpublished data.
Received 10/28/99; revised 2/18/00; accepted 2/18/00.
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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 C. A. Kruse, S. Visonneau, B. K. Kleinschmidt-DeMasters, C. J. Gup, G. G. Gomez, D. B. Paul, and D. Santoli The Human Leukemic T-Cell Line, TALL-104, Is Cytotoxic to Human Malignant Brain Tumors and Traffics through Brain Tissue: Implications for Local Adoptive Immunotherapy Cancer Res., October 1, 2000; 60(20): 5731 - 5739. [Abstract] [Full Text]
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