| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
Clinical Trials |
-Galactosylceramide (KRN7000) in Patients with Solid Tumors
Departments of Medical Oncology [G. G., R. R., N. N., H. J. J. v.d. V., A. J. M. v. d. E., H. M. P.] and Pathology [B. M. E. v. B., R. J. S.], Vrije Universiteit Medical Center, HV 1081 Amsterdam, the Netherlands; Department of Medical Oncology, University Hospital Nijmegen St. Radboud, the Netherlands [C. J. A. P., M. P.]; Kirin Brewery Co. Ltd., Tokyo, Japan [Y. A., N. N.]; NDDO Oncology, Amsterdam, the Netherlands [M. R.]; Slotervaart Hospital, Amsterdam, the Netherlands [J. B.]; and Innsbruck University, Innsbruck, Austria [H. Z.]
 |
ABSTRACT |
|---|
 Top ABSTRACT INTRODUCTIONPATIENTS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
-galactosylceramide (KRN7000) is a glycosphingolipid that has been shown to inhibit tumor growth and to prolong survival in inoculated mice through activation of natural killer (NK) T cells. We performed a dose escalation study of KRN7000 in advanced cancer patients. Experimental Design: Patients with solid tumors received i.v. KRN7000 (50–4800 µg/m2) on days 1, 8, and 15 of a 4-weekly cycle. Patients were given 1 cycle and, in the absence of dose-limiting toxicity or progression, treatment was continued. Pharmacokinetics (PK) and immunomonitoring were performed in all patients.
Results: Twenty-four patients were entered into this study. No dose-limiting toxicity was observed over a wide range of doses (50–4800 µg/m2). PK was linear in the dose range tested. Immunomonitoring demonstrated that NKT cells (CD3+V24+Vß11+) typically disappeared from the blood within 24 h of KRN7000 injection. Additional biological effects included increased serum cytokine levels (tumor necrosis factor and granulocyte macrophage colony-stimulating factor) in 5 of 24 patients and a transient decrease in peripheral blood NK cell numbers and cytotoxicity in 7 of 24 patients. Importantly, the observed biological effects depended on pretreatment NKT-cell numbers rather than on the dose of KRN7000. Pretreatment NKT-cell numbers were significantly lower in patients compared with healthy controls (P = 0.0001). No clinical responses were recorded and seven patients experienced stable disease for a median duration of 123 days.
Conclusion: i.v. KRN7000 is well tolerated in cancer patients over a wide range of doses. Biological effects were observed in several patients with relatively high pretreatment NKT-cell numbers. Other therapeutic strategies aiming at reconstitution of the deficient NKT-cell population in cancer patients may be warranted.
|
INTRODUCTION |
|---|
|
TopABSTRACTINTRODUCTIONPATIENTS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
-GalCer3
(KRN7000) is a synthetic glycolipid, originally extracted from the marine sponge Agelas mauritianus. KRN7000 consists of a galactose and a ceramide molecule linked in an -configuration (Fig. 1
; Ref. 1
). On the basis of structure-activity relationship studies, KRN7000 was selected as the most potent agent of a series of compounds (2)
.
|
and IL-4, thereby activating other effector cells including NK cells, T cells, B cells, and macrophages (10, 11, 12)
. The administration of -GalCer in animals also induced the production of other cytokines, such as IL-2, IL-3, IL-6, IL-12, GM-CSF, and stem cell factor, which may also reinforce the immune system (13)
.
In in vivo animal studies, KRN7000 showed antitumor effects against various tumors (including melanoma, sarcoma, colon carcinoma, and lymphoma) in hepatic and lung metastases models (14
, 15)
. Repeated administration with intervals was superior to single injections. Interestingly, apart from microgranulomas in liver and spleen in the highest dose level, no side effects were reported in mice, rats, and monkeys with doses up to 2200 µg/kg, 440 µg/kg, and 660 µg/kg, respectively, given for 28 consecutive days.4
The administration of -GalCer (100 µg/kg) in mice resulted in a transient increase in alanine aminotransferase activity that was found to be mediated by activated NKT cells (16
, 17)
. Antitumor activity has been observed in mouse models at doses ranging from 0.01 to 100 µg/kg (14
, 15
, 18)
.
We performed a Phase I study of KRN7000 given weekly i.v. for 3 consecutive weeks to patients with refractory solid tumors. The PK and biological effects of KRN7000 were monitored. The major end point of this study was to identify the MTD and the OBAD of KRN7000.
|
PATIENTS AND METHODS |
|---|
|
TopABSTRACTINTRODUCTIONPATIENTS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
On-Study Evaluations.
History and physical examination were taken at study entry and before every cycle of treatment. Hematology, chemistries and urinalysis, electrocardiogram, and chest X-ray were also assessed before study entry. Proper diagnostic techniques were used to adequately assess the tumor extension. An ACTH test (synachten, 250 µg i.v.; blood sampling for serum cortisol were taken immediately before the injection and 30 min thereafter) was performed, and antithyroid antibodies were measured at study entry. Chemistries and full blood cell counts were repeated every week, and radiological assessment of tumor response was performed every two cycles.
Study Design.
This was an open-label nonrandomized dose-escalation study. The aims of the study were to evaluate the safety profile of KRN7000, to determine the MTD; to study the biological and immunological changes induced by KRN7000, and to identify an OBAD. Furthermore, drug concentrations in blood and urine and possible antitumor activity were investigated over time. Toxicity was assessed using the National Cancer Institute Common Toxicity Criteria (CTC), and antitumor activity according to the WHO criteria. The MTD was defined as a dose associated with dose-limiting toxicity in at least two of six patients. Dose-limiting toxicity was defined as any adverse drug reaction (ADR) of 
CTC grade 3, excluding nausea, vomiting, and grade 3 fever or any ADR that caused interruption of the weekly schedule administration. OBAD was to be defined on the basis of results obtained by immunological monitoring.
At least three patients were entered on each dose level, and increased to six if dose-limiting toxicities were encountered. Expansion of one or more cohorts was also planned in case of identification of biological activities.
Drug Formulation and Administration.
The clinical dosage form of KRN7000 was supplied as vials containing lyophilized KRN7000 (0.2 mg), sucrose (56 mg), L-histidine (7.5 mg), and polysorbate 20 (5 mg) and was stored at 2°C to 8°C. Just before administration, KRN7000 was reconstituted with 1 ml of water. KRN7000 was administered by slow i.v. infusion on days 1, 8, and 15 of a 4-weekly cycle. Before drug administration, signs and symptoms of toxicity had to have disappeared. Doses administered and number of patients included at each dose level are summarized in Table 1
.
|
Urines were collected in 24-h aliquots both before and after drug administration. All of the samples were stored at -20°C until analysis. KRN7000 concentrations in urine and serum were determined with 13C-labeled KRN7000 as an internal standard using validated high-performance liquid chromatography tandem mass spectrometry. After thawing, urine, to which 10% Tween 20 was added, and serum samples were mixed with an internal standard in phosphate buffer with 1 M NaOH. Then KRN7000 and an internal standard in samples were extracted twice into ethylacetate. The organic phase was evaporated after phase separation. The residue was reconstituted with methanol, and the aliquot was injected into a liquid chromatography system with a tandem mass spectrometry, either a Quattro II (Micromass, Manchester, United Kingdom) or an API 365 (PE Sciex, Foster City, CA), with electrospray, by which product ions, galactose of KRN7000 and 13C-labeled KRN7000, from deprotonated ions were detected.
Immunomonitoring.
To assess biological activities of KRN7000, extensive immunomonitoring was performed, including phenotyping of PBMCs, NK cell lytic activity, stimulation of the MLR, and serum cytokine levels. A total of 20 ml of heparinized PB were collected before the first administration (day 1), and on days 2, 3, 5, and 8, then before the third administration (day 15) and on days 16, 19, and 22, to perform cellular assays. Serum samples for cytokine evaluation were stored at -20° before, and 1, 2, 4, 6, 8, 10, and 24 h after, the first KRN7000 injection and before, and 2, 4, 10, and 24 h after, the third injection.
Preparation of PBMCs for Cryopreservation.
PBMCs were isolated from heparinized diluted PB by Lymphoprep (Nycomed Pharma, Oslo, Norway) density gradient centrifugation. Cells were washed and resuspended at 4°C at 6 x 106 viable cells/ml in RPMI 1640 (BioWhittaker, Verviers, Belgium) containing 25% FCS and 12.5% DMSO. Cells were then cryopreserved and stored in liquid nitrogen until use, allowing for simultaneous testing of all of the PBMC samples from each individual patient.
PBMC Proliferation Assay.
Thawed PBMCs were cultured in Iscove‘s modified Dulbecco‘s medium (BioWhittaker) containing 20% heat-inactivated HPS (CLB, Amsterdam, the Netherlands). Two hundred thousand PBMCs were cultured in the presence of 0, 1, 10, and 100 ng/ml KRN7000 (150 µl/well) for 7 days at 37°C with 5% CO2 in humidified air. Then [3H]thymidine (0.4 µCi; Amersham Pharmacia Biotech, Roosendaal, the Netherlands) was added to each well, and cells were incubated for an additional 18 h. Cells were harvested (Skatron micro96 harvester; Skatron Instruments, Lier, Norway), and radioactivity was measured by a beta counter (Microbeta counter; Wallac Inc., Gaithersburg, MD).
Allogeneic MLR Assay.
Thawed PBMCs of the patients were used as allogeneic stimulatory cells for healthy responder PBMCs in the MLR. For each patient, optimal responder PBMCs were first selected from a panel of three donors. Triplicate cultures were set up with 0.25 x 104, 1.0 x 104, and 4.0 x 104 irradiated (30 Gy) stimulator cells and 2 x 105 responder cells in Iscove‘s modified Dulbecco‘s medium containing 10% HPS in a 96-well round-bottomed plate (Corning Inc., Corning, NY). After 5 days of culture, lymphocyte proliferation was measured as [3H]thymidine uptake as described above.
Flowcytometric Cell Surface Analysis.
Antibodies used in this study were: FITC-labeled anti-CD1c (Ancell, Bayport, MN); RPE-Cy5-labeled anti-CD3 (Dako, Carpinteria, CA); FITC-labeled anti-CD4 (Becton Dickinson & Co., Franklin Lakes, NJ); PE-labeled anti-CD8 (Becton Dickinson); RPE-Cy5-labeled anti-CD14 (Coulter, Fullerton, CA); PE-labeled anti-CD56 (Becton Dickinson); PE-labeled anti-CD86 (PharMingen); FITC-labeled anti-V24 (Immunotech, Marseille, France); PE-labeled anti-Vß11 (Immunotech); and FITC-labeled anti-HLA-DR (Becton Dickinson). Isotype controls used: were FITC-labeled mIgG1 (Becton Dickinson); PE-labeled mIgG1 (Becton Dickinson); RPE-Cy5-labeled mIgG1 (DAKO); FITC-labeled mIgG2a (Becton Dickinson); and RPE-Cy5-labeled mIgG2a (Coulter). All of the samples from a patient were tested in the same run and one vial of a large batch of control cells was thawed and stained simultaneously in every experiment. Thawed cells were suspended in PBS and fixed with 1% paraformaldehyde. After washing, cells were resuspended in PBS containing, as blocking agent, 10% HPS, mIgG1 (1:10 dilution) and mIgG2a (1:10 dilution) for 30 min at room temperature. After washing, antibodies and isotype controls were added to the cell pellets. Cells were kept in darkness at 4°C until analysis. Analysis was done using a FACStar plus (Becton Dickinson).
NK Cell Cytotoxicity Assay.
PBMCs were tested for NK cell cytotoxic activity in a 51Cr release assay. After thawing, PBMCs (0.5 x 106 cells/ml in RPMI 1640 with 10% heat inactivated FCS were precultured overnight to recover. NK-sensitive target cells (K562) were labeled with 100 µCi Na2[51Cr]O4 (Amersham Pharmacia Biotech) at 37°C for 1 h. 51Cr-labeled K562 cells were washed three times and resuspended at 0.1 x 106 cells/ml in RPMI 1640 with 10% FCS. PBMCs and 51Cr-labeled target cells (1 x 104 cells/well) were added to 96-wells round-bottomed plates at E:T ratios of 10:1, 20:1, and 40:1 (triplicate cultures). Additional wells were set up for the measurement of spontaneous and total [51Cr] release by adding target cells to wells with 100 µl of medium or 1% Triton X-100, respectively. Plates were centrifuged (150 x g) for 1 min and then incubated at 37°C with 5% CO2 for 4 h. One hundred µl of supernatant were harvested from each well and transferred to a 96-well sample plate (Beta plate, Wallac Inc.), 100 µl of scintillation fluid were added, and plates were shaken for at least 15 min. Radioactivity was measured by a beta counter. The percentage of specific 51Cr release was calculated as follows: [(test cpm - spontaneous cpm)/(total cpm - spontaneous cpm)] x 100. A large batch of frozen healthy PBMCs were used in all of the experiments to evaluate interassay variation.
Serum Cytokine Measurement.
All of the samples were kept at -80°C until testing. IL-1ß, IL-2, IL-4, IL-10, IL-12, IFN-, GM-CSF, and TNF- in serum were measured using ELISA kits according to the manufacturer’s instructions. The IL-2 ELISA was purchased from Cell Sciences Inc. (Diaclone; Norwood, MA) and all of the other ELISAs (high sensitive kits except for IFN-) from R&D Systems Europe Ltd. (Abingdon, United Kingdom).
|
RESULTS |
|---|
|
TopABSTRACTINTRODUCTIONPATIENTS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
. On all dose levels, three patients were entered, with the exception of dose level 1, in which six patients were entered because distinct biological changes were seen in this first cohort of patients.
|
In conclusion, no serious drug-related adverse event occurred during this study. The ACTH test was not negatively affected in the patients who were tested again after therapy discontinuation. Further dose escalation could not be performed using the current drug formulation; therefore, the MTD could not be identified.
Immunological Monitoring.
Pretreatment, KRN7000-induced in vitro lymphocyte proliferation was observed in 4 of 24 patients (doubling of cpm counts induced by at least one dose of KRN7000): in 2 with high NKT cells at the start and 2 with low NKT cells (see NKT cells below). There were no consistent KRN7000-induced changes in the numbers of granulocytes, monocytes, B cells, and CD4+ or CD8+ T cells. Effects on the expression of HLA-DR, CD86, and CD1c on either CD14-positive or CD14-negative nonlymphocytic (based on light scatter) PBMCs were also inconsistent. Moreover, there were no KRN7000-related effects on the stimulatory capacity in MLR assays.
NKT Cells.
NKT cells, expressing the invariant T-cell receptor consisting of a V24 chain paired with a Vß11 chain (8)
, were measured in the PB of all of the patients except in two of the first- and one of the second-dose level. The number of circulating NKT cells in cancer patients was significantly lower than that of healthy volunteers (Fig. 2
; P = 0.0001, Wilcoxon two-sample test). Using the median number of NKT cells in patients (333 cells/ml PB) as the cutoff value, we defined an NKT-high group and an NKT-low group (Fig. 2)
. The NKT cells in the PB in the NKT-high group decreased to undetectable levels within 24 h after the first administration of KRN7000 at all of the dose levels, and recovery to the preadministration levels was not observed within a week (Fig. 3)
.
|
|
and IL-12, both of which can trigger NK cells, increased when KRN7000 was administered to mice (18)
. Here, only one patient (patient 3 on the first cohort), who had a relatively high pretreatment NKT cell count, showed a faint but detectable increase in serum IFN- at 6 h after the first KRN7000 administration (Fig. 4A)
, and an increase in serum IL-12 peaking 8 h after KRN7000 administration (Fig. 4B)
. Detectable levels of IFN- or IL-12 (>15 or 0.8 pg/ml, respectively) were never observed in the NKT-low group (data not shown).
|
levels, peaking 4–6 h after KRN7000 injection (Fig. 5, A and C)
. Of these, patient 3, who experienced severe fever after the first administration of KRN7000, besides showing both IL-12 and IFN- production, also developed the highest levels of GM-CSF and TNF-. In contrast, in the NKT-low group, only slight fluctuations of GM-CSF and TNF- serum levels were seen after KRN7000 administration (Fig. 5, B and D)
.
|
NK Cells.
The number of NK cells as well as the NK cytotoxicity of the NKT-high group are shown in Fig. 6
. Seven of 10 patients with high pretreatment NKT cell numbers showed a transient decrease in NK cell numbers and NK cell cytolytic activity 24 h after KRN7000 treatment. No apparent response in NK cells was observed in patients in the NKT-low group (data not shown).
|
PKs.
Table 3
summarizes the major PK parameters. There was no indication of drug accumulation or saturation in the serum PK. Of note, plasma concentrations associated with activity in mice (18)
were already achieved at dose level 3. There was a linear correlation between dose administered and area under the curve (AUC). Fig. 7
shows the serum PK at all of the dose levels. No apparent drug accumulation was observed after multiple dosing. KRN7000 was not detectable in urine at any dose level.
|
|
|
DISCUSSION |
|---|
|
TopABSTRACTINTRODUCTIONPATIENTS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
-GalCer, with potent tumor-inhibitory activity in metastatic tumor models in mice (1
, 2
, 14
, 15
, 18) . In the dose range tested, spanning almost a 100-fold range in doses, we did not observe any dose-limiting toxicity. No accumulation was observed with weekly administrations, and urine excretion of KRN7000 was essentially under the detection limit of our method. In only one patient, treated at the lowest-dose level of 50 µg/m2, did we observe occurrence of high fever, with shivering, which was clearly drug related. In this patient, we also detected increased serum levels of IFN-, IL-12, GM-CSF, and TNF-. Although cytokine production was infrequently observed in the study, this was limited to patients with relatively high circulating NKT-cell numbers before treatment. NKT cells have been shown to play an important role in antitumor immune responses. A protective role of NKT cells in tumor-immunosurveillance was demonstrated (19) , and NKT cells were reported to be essential for the antitumor effects of IL-12 (20) . The antitumor effects of KRN7000 are crucially dependent on NKT cells, because no antitumor activity was observed in NKT cell-deficient mice (9) . Although KRN7000-activated NKT cells are able to directly lyse a wide variety of tumor cell lines (9 , 21 , 22) , they also enhance antitumor immune responses by rapidly releasing large amounts of cytokines that secondarily activate NK cells, T cells, B cells, dendritic cells, and monocytes (10, 11, 12 , 14 , 23) . Clinical benefit for cancer patients was anticipated, because both human and mouse CD1d can present KRN7000 to NKT cells from either species, illustrating a strong conservation of recognition (5 , 6) .
An important finding of our study was the observation that the initial size of the circulating NKT-cell pool in cancer patients was significantly lower compared with that in healthy controls. This was previously observed in patients with melanoma and prostate cancer (21
, 24)
, but our data suggest that the decrease is more widespread among cancer patients. Although the mechanism of this decrease has not been investigated, it could be caused by tumor-derived immunosuppressive factors or the intra- or peritumoral accumulation of NKT cells. Alternatively, the size of the NKT-cell population could have been reduced before tumor development, thereby acting as a risk factor. Another important finding was the observation that (residual) PB NKT cells rapidly disappeared from the circulation when KRN7000 was administered. Interestingly, a similar KRN7000-induced depletion of NKT cells from liver, spleen, and lymph nodes has been reported in mice and has been attributed to apoptosis after NKT-cell activation (and subsequent cytokine production; Refs. 11
, 17
, 25
). The exact duration of the NKT-cell depletion after KRN7000 administration was not investigated in our study, but NKT-cell repopulation was not observed in the week after the injection of KRN7000. The 1-week intervals for this study were based on the best antitumor activity obtained by i.v. administration of KRN7000 on days 1, 5, and 9 in mice (15)
. However, weekly i.v. injections may well be too frequent because they may suppress NKT-cell repopulation. This is supported by our observation that individuals showing increased serum levels of GM-CSF and TNF- in response to KRN7000 showed this response only after the first administration, when relatively high numbers of NKT cells were still present in the circulation. Interestingly, because human NKT cells were indeed shown to produce GM-CSF and TNF- on activation in vitro, a role for KRN7000-activated NKT cells in the recruitment and maturation of dendritic cells has been inferred (26)
.
In mice, NKT cells were reported to rapidly activate NK cells on i.v. administration of KRN7000 (10) . Other studies demonstrated that KRN7000 administration resulted in an increase in NK cell numbers and cytotoxicity in the liver (11 , 17) . Here, we could not identify any sign of NK cell activation or an increase in NK cell lytic activity on KRN7000 administration; actually, in the majority of patients with high pretreatment NKT cell numbers, a decrease in NK cell cytotoxicity was observed 24 h after KRN7000 administration, whereas no apparent change was observed in patients with low NKT cell counts. Although this decrease in PB NK cell numbers and cytotoxicity, at first glance, contrasts with murine studies, it should be noted that a temporary loss of NK cells in the spleen, accompanied the KRN7000, induced an increase in liver NK cells in mice (17) . Recently, KRN7000 was shown to enhance the in vitro tumor-killing activity of human hepatic CD56+ NK cells, underscoring the strong functional similarities in function between mouse and human NKT cells (27) .
No clinical responses were observed in our study, although a number of patients had relatively long-lasting stable disease. The absence of clinical responses in combination with the low frequency of patients in which signs of immunological activation were observed, may suggest that this schedule and/or route of administration should not be further investigated in a Phase II study. The low number of NKT cells in cancer patients most likely contributed to the lack of immunoreactivity after KRN7000 injections. This view is supported by the fact that immune activation was observed in only those patients who had relatively high pretreatment NKT-cell numbers. An intriguing observation was, however, that even at the lowest dose tested (50 µg/m2), cytokine production and NKT-cell depletion were observed. This may support the study of lower doses than those tested in our trial. However the fact that -GalCer inhibits the development of autoimmune diabetes (28
, 29)
and multiple sclerosis (30, 31, 32)
in mice, and that the elimination of CD1-dependent NKT cells has been shown to improve tumor rejection (33)
, adds further complexity, which suggests that additional preclinical studies are needed to allow optimal clinical development of this compound.
In conclusion, this Phase I study of KRN7000 did not reach the MTD and did not record substantial toxicities across a broad range of doses. Moreover, no OBAD was defined, despite a very intensive immunological monitoring in all of the patients included in the study. However, the strong, and evolutionarily highly conserved, similarities between mouse and human NKT cells warrant intensive research into alternative approaches and routes of administration (e.g., s.c.), to try to promote KRN7000-induced antitumor effects in humans. Among these are adoptive transfer of NKT cells and the treatment of cancer patients with KRN7000-pulsed dendritic cells; both strategies have resulted in strong antitumor immune responses in mice (34 , 35) .
|
ACKNOWLEDGMENTS |
|---|
|
FOOTNOTES |
|---|
1 To whom requests for reprints should be addressed, at Vrije Universiteit Medical Center, Division of Medical Oncology, De Boelelaan 1117, HV 1081 Amsterdam, the Netherlands. Phone: 31-20-4444321; Fax: 31-20-4444079; E-mail: G.Giaccone{at}vumc.nl 
2 EORTC Biological Therapeutics Development Group.
3 The abbreviations used are: -GalCer, -galactosylceramide; NK, natural killer; NKT, NK T (cell); TNF, tumor necrosis factor; GM-CSF, granulocyte macrophage colony-stimulating factor; IL, interleukin; PK, pharmacokinetic(s); MTD, maximum tolerated dose; OBAD, optimal biologically active dose; ACTH, adrenocorticotropic hormone; PB, peripheral blood; PBMC, PB mononuclear cell; MLR, mixed lymphocyte reaction; HPS, human pooled serum; RPE, R-phyco erythrin.
4 G. Giaccone et al., unpublished results.
Received 3/29/02; revised 8/ 8/02; accepted 8/20/02.
|
REFERENCES |
|---|
|
TopABSTRACTINTRODUCTIONPATIENTS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
-galactosylceramides against B16-bearing mice. J. Med. Chem., 38: 2176-2187, 1995.[CrossRef][Medline]
14 NKT cells by glycosylceramides. Science (Wash. DC), 278: 1626-1629, 1997.24+ CD4-CD8- T cells. J. Exp. Med., 186: 109-120, 1997.-galactosylceramide by natural killer T cells is highly conserved through mammalian evolution. J. Exp. Med., 188: 1521-1528, 1998. upon activation by anti-CD3 or CD1. J. Immunol., 159: 2240-2249, 1997.14 NKT cells. Proc. Natl. Acad. Sci. USA, 95: 5690-5693, 1998. and NK cells, but not perforin-mediated cytotoxicity, to anti-metastatic effect of -galactosylceramide. Eur. J. Immunol., 31: 1720-1727, 2001.[CrossRef][Medline]
-galactosylceramide (AGL-517) treatment protects mice from lethal irradiation. Exp. Hematol., 25: 935-944, 1997.[Medline]
-galactosylceramide. KRN7000. Cancer Res., 58: 1202-1207, 1998.-galactosylceramide in mice. J. Immunol., 166: 6578-6584, 2001.-galactosylceramide. Eur. J. Immunol., 30: 1919-1928, 2000.[CrossRef][Medline]
-galactosylceramide. KRN7000, in mice with EL-4 hepatic metastasis and its cytokine production. Oncol. Res., 10: 561-568, 1998.[Medline]
14 NKT cells in IL-12-mediated rejection of tumors. Science (Wash. DC), 278: 1623-1626, 1997.24 NKT cells. Cancer Res, 59: 5102-5105, 1999.-galactosylceramide demonstrates its immunopotentiating effect by inducing interleukin (IL)-12 production by dendritic cells and IL-12 receptor expression on NKT cells. J. Exp. Med., 189: 1121-1128, 1999. production by invariant NK T cells in advanced cancer. J. Immunol., 167: 4046-4050, 2001.24JQ T-cells: a feedback mechanism for immune regulation. J. Immunol., 165: 3759-3762, 2000.
-glycosylceramides enhance the antitumor cytotoxicity of hepatic lymphocytes obtained from cancer patients by activating CD3-CD56+ NK cells in vitro. J. Immunol., 165: 1659-1664, 2000.-galactosylceramide treatment prevents the onset of autoimmune type I diabetes. Nat. Med., 7: 1057-1062, 2001.[CrossRef][Medline]
-galactosylceramide prevents autoimmune diabetes in non-obese diabetic mice. Nat. Med., 7: 1052-1056, 2001.[CrossRef][Medline]
-galactosylceramide. J. Immunol., 163: 2387-2391, 1999.14 NKT cells. Int. J. Cancer, 91: 523-528, 2001.[CrossRef][Medline]This article has been cited by other articles:
|
|
 |
|
  D. Bagnara, A. Ibatici, M. Corselli, N. Sessarego, C. Tenca, A. De Santanna, A. Mazzarello, A. Daga, R. Corvo, G. De Rossi, et al. Adoptive immunotherapy mediated by ex vivo expanded natural killer T cells against CD1d-expressing lymphoid neoplasms Haematologica, July 1, 2009; 94(7): 967 - 974. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. H. Tripp, F. Sparber, I. F. Hermans, N. Romani, and P. Stoitzner Glycolipids Injected into the Skin Are Presented to NKT Cells in the Draining Lymph Node Independently of Migratory Skin Dendritic Cells J. Immunol., June 15, 2009; 182(12): 7644 - 7654. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Lindqvist, J. Persson, K. Thorn, and A. M. Harandi The Mucosal Adjuvant Effect of {alpha}-Galactosylceramide for Induction of Protective Immunity to Sexually Transmitted Viral Infection J. Immunol., May 15, 2009; 182(10): 6435 - 6443. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Bricard, V. Cesson, E. Devevre, H. Bouzourene, C. Barbey, N. Rufer, J. S. Im, P. M. Alves, O. Martinet, N. Halkic, et al. Enrichment of Human CD4+ V{alpha}24/V{beta}11 Invariant NKT Cells in Intrahepatic Malignant Tumors J. Immunol., April 15, 2009; 182(8): 5140 - 5151. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H.-J. Choi, H. Xu, Y. Geng, A. Colmone, H. Cho, and C.-R. Wang Bacterial infection alters the kinetics and function of iNKT cell responses J. Leukoc. Biol., December 1, 2008; 84(6): 1462 - 1471. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Moreno, J. W. Molling, S. von Mensdorff-Pouilly, R. H. M. Verheijen, E. Hooijberg, D. Kramer, A. W. Reurs, A. J. M. van den Eertwegh, B. M. E. von Blomberg, R. J. Scheper, et al. IFN-{gamma}-Producing Human Invariant NKT Cells Promote Tumor-Associated Antigen-Specific Cytotoxic T Cell Responses J. Immunol., August 15, 2008; 181(4): 2446 - 2454. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Biburger and G. Tiegs Activation-induced NKT cell hyporesponsiveness protects from {alpha}-galactosylceramide hepatitis and is independent of active transregulatory factors J. Leukoc. Biol., July 1, 2008; 84(1): 264 - 279. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Ito, K. Ando, T. Ishikawa, T. Nakayama, M. Taniguchi, K. Saito, M. Imawari, H. Moriwaki, T. Yokochi, S. Kakumu, et al. Role of V{alpha}14+ NKT cells in the development of Hepatitis B virus-specific CTL: activation of V{alpha}14+ NKT cells promotes the breakage of CTL tolerance Int. Immunol., July 1, 2008; 20(7): 869 - 879. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. A. Berzofsky and M. Terabe NKT Cells in Tumor Immunity: Opposing Subsets Define a New Immunoregulatory Axis J. Immunol., March 15, 2008; 180(6): 3627 - 3635. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Torres, C. Paget, J. Fontaine, T. Mallevaey, T. Matsuoka, T. Maruyama, S. Narumiya, M. Capron, P. Gosset, C. Faveeuw, et al. Prostaglandin D2 Inhibits the Production of IFN-{gamma} by Invariant NK T Cells: Consequences in the Control of B16 Melanoma J. Immunol., January 15, 2008; 180(2): 783 - 792. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Wu and L. V. Kaer Role of NKT Cells in the Digestive System. II. NKT cells and diabetes Am J Physiol Gastrointest Liver Physiol, November 1, 2007; 293(5): G919 - G922. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Shimizu, Y. Kurosawa, M. Taniguchi, R. M. Steinman, and S.-i. Fujii Cross-presentation of glycolipid from tumor cells loaded with {alpha}-galactosylceramide leads to potent and long-lived T cell mediated immunity via dendritic cells J. Exp. Med., October 29, 2007; 204(11): 2641 - 2653. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
O. Adotevi, B. Vingert, L. Freyburger, P. Shrikant, Y.-C. Lone, F. Quintin-Colonna, N. Haicheur, M. Amessou, A. Herbelin, P. Langlade-Demoyen, et al. B Subunit of Shiga Toxin-Based Vaccines Synergize with {alpha}-Galactosylceramide to Break Tolerance against Self Antigen and Elicit Antiviral Immunity J. Immunol., September 1, 2007; 179(5): 3371 - 3379. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Ebensen, C. Link, P. Riese, K. Schulze, M. Morr, and C. A. Guzman A Pegylated Derivative of {alpha}-Galactosylceramide Exhibits Improved Biological Properties J. Immunol., August 15, 2007; 179(4): 2065 - 2073. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. W.L. Teng, J. A. Westwood, P. K. Darcy, J. Sharkey, M. Tsuji, R. W. Franck, S. A. Porcelli, G. S. Besra, K. Takeda, H. Yagita, et al. Combined Natural Killer T-Cell Based Immunotherapy Eradicates Established Tumors in Mice Cancer Res., August 1, 2007; 67(15): 7495 - 7504. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y.-J. Chang, J.-R. Huang, Y.-C. Tsai, J.-T. Hung, D. Wu, M. Fujio, C.-H. Wong, and A. L. Yu Potent immune-modulating and anticancer effects of NKT cell stimulatory glycolipids PNAS, June 19, 2007; 104(25): 10299 - 10304. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. J.J. van derVliet, H. B. Koon, S. C. Yue, B. Uzunparmak, V. Seery, M. A. Gavin, A. Y. Rudensky, M. B. Atkins, S. P. Balk, and M. A. Exley Effects of the Administration of High-Dose Interleukin-2 on Immunoregulatory Cell Subsets in Patients with Advanced Melanoma and Renal Cell Cancer Clin. Cancer Res., April 1, 2007; 13(7): 2100 - 2108. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. F. Hermans, J. D. Silk, U. Gileadi, S. H. Masri, D. Shepherd, K. J. Farrand, M. Salio, and V. Cerundolo Dendritic Cell Function Can Be Modulated through Cooperative Actions of TLR Ligands and Invariant NKT Cells J. Immunol., March 1, 2007; 178(5): 2721 - 2729. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. M. Coquet, K. Kyparissoudis, D. G. Pellicci, G. Besra, S. P. Berzins, M. J. Smyth, and D. I. Godfrey IL-21 Is Produced by NKT Cells and Modulates NKT Cell Activation and Cytokine Production J. Immunol., March 1, 2007; 178(5): 2827 - 2834. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Yanagisawa, M. A. Exley, X. Jiang, N. Ohkochi, M. Taniguchi, and K.-i. Seino Hyporesponsiveness to Natural Killer T-Cell Ligand {alpha}-Galactosylceramide in Cancer-Bearing State Mediated by CD11b+ Gr-1+ Cells Producing Nitric Oxide Cancer Res., December 1, 2006; 66(23): 11441 - 11446. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Hong, H. Lee, M. Oh, C.-Y. Kang, S. Hong, and S.-H. Park CD4+ T Cells in the Absence of the CD8+ Cytotoxic T Cells Are Critical and Sufficient for NKT Cell-Dependent Tumor Rejection J. Immunol., November 15, 2006; 177(10): 6747 - 6757. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. J.J. van der Vliet, S. P. Balk, and M. A. Exley Natural Killer T Cell-Based Cancer Immunotherapy. Clin. Cancer Res., October 15, 2006; 12(20): 5921 - 5923. [Full Text] [PDF]
|
|
|
|
|
|
S. Motohashi, A. Ishikawa, E. Ishikawa, M. Otsuji, T. Iizasa, H. Hanaoka, N. Shimizu, S. Horiguchi, Y. Okamoto, S.-i. Fujii, et al. A Phase I Study of In vitro Expanded Natural Killer T Cells in Patients with Advanced and Recurrent Non-Small Cell Lung Cancer Clin. Cancer Res., October 15, 2006; 12(20): 6079 - 6086. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Shimizu, M. Hidaka, N. Kadowaki, N. Makita, N. Konishi, K. Fujimoto, T. Uchiyama, F. Kawano, M. Taniguchi, and S.-i. Fujii Evaluation of the Function of Human Invariant NKT Cells from Cancer Patients Using {alpha}-Galactosylceramide-Loaded Murine Dendritic Cells. J. Immunol., September 1, 2006; 177(5): 3484 - 3492. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Lizee, L. G. Radvanyi, W. W. Overwijk, and P. Hwu Improving Antitumor Immune Responses by Circumventing Immunoregulatory Cells and Mechanisms. Clin. Cancer Res., August 15, 2006; 12(16): 4794 - 4803. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. R. Brutkiewicz CD1d Ligands: The Good, the Bad, and the Ugly J. Immunol., July 15, 2006; 177(2): 769 - 775. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. H. Meyer, S. Goya, O. Akbari, G. J. Berry, P. B. Savage, M. Kronenberg, T. Nakayama, R. H. DeKruyff, and D. T. Umetsu Glycolipid activation of invariant T cell receptor+ NK T cells is sufficient to induce airway hyperreactivity independent of conventional CD4+ T cells PNAS, February 21, 2006; 103(8): 2782 - 2787. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Liu, J. Idoyaga, A. Charalambous, S.-i. Fujii, A. Bonito, J. Mordoh, R. Wainstok, X.-F. Bai, Y. Liu, and R. M. Steinman Innate NKT lymphocytes confer superior adaptive immunity via tumor-capturing dendritic cells J. Exp. Med., December 5, 2005; 202(11): 1507 - 1516. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. P. Uldrich, N. Y. Crowe, K. Kyparissoudis, D. G. Pellicci, Y. Zhan, A. M. Lew, P. Bouillet, A. Strasser, M. J. Smyth, and D. I. Godfrey NKT Cell Stimulation with Glycolipid Antigen In Vivo: Costimulation-Dependent Expansion, Bim-Dependent Contraction, and Hyporesponsiveness to Further Antigenic Challenge J. Immunol., September 1, 2005; 175(5): 3092 - 3101. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.-Y. Ko, H.-J. Ko, W.-S. Chang, S.-H. Park, M.-N. Kweon, and C.-Y. Kang {alpha}-Galactosylceramide Can Act As a Nasal Vaccine Adjuvant Inducing Protective Immune Responses against Viral Infection and Tumor J. Immunol., September 1, 2005; 175(5): 3309 - 3317. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Haraguchi, T. Takahashi, A. Matsumoto, T. Asai, Y. Kanda, M. Kurokawa, S. Ogawa, H. Oda, M. Taniguchi, H. Hirai, et al. Host-Residual Invariant NK T Cells Attenuate Graft-versus-Host Immunity J. Immunol., July 15, 2005; 175(2): 1320 - 1328. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Matsuda, T. Suda, J. Sato, T. Nagata, Y. Koide, K. Chida, and H. Nakamura {alpha}-Galactosylceramide, a Ligand of Natural Killer T Cells, Inhibits Allergic Airway Inflammation Am. J. Respir. Cell Mol. Biol., July 1, 2005; 33(1): 22 - 31. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Ota, K. Takeda, H. Akiba, Y. Hayakawa, K. Ogasawara, Y. Ikarashi, S. Miyake, H. Wakasugi, T. Yamamura, M. Kronenberg, et al. IFN-{gamma}-mediated negative feedback regulation of NKT-cell function by CD94/NKG2 Blood, July 1, 2005; 106(1): 184 - 192. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. J. Smyth, M. E. Wallace, S. L. Nutt, H. Yagita, D. I. Godfrey, and Y. Hayakawa Sequential activation of NKT cells and NK cells provides effective innate immunotherapy of cancer J. Exp. Med., June 20, 2005; 201(12): 1973 - 1985. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. H. Chang, K. Osman, J. Connolly, A. Kukreja, J. Krasovsky, M. Pack, A. Hutchinson, M. Geller, N. Liu, R. Annable, et al. Sustained expansion of NKT cells and antigen-specific T cells after injection of {alpha}-galactosyl-ceramide loaded mature dendritic cells in cancer patients J. Exp. Med., May 2, 2005; 201(9): 1503 - 1517. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. S. Bezbradica, A. K. Stanic, N. Matsuki, H. Bour-Jordan, J. A. Bluestone, J. W. Thomas, D. Unutmaz, L. Van Kaer, and S. Joyce Distinct Roles of Dendritic Cells and B Cells in Va14Ja18 Natural T Cell Activation In Vivo J. Immunol., April 15, 2005; 174(8): 4696 - 4705. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Ishikawa, S. Motohashi, E. Ishikawa, H. Fuchida, K. Higashino, M. Otsuji, T. Iizasa, T. Nakayama, M. Taniguchi, and T. Fujisawa A Phase I Study of {alpha}-Galactosylceramide (KRN7000)-Pulsed Dendritic Cells in Patients with Advanced and Recurrent Non-Small Cell Lung Cancer Clin. Cancer Res., March 1, 2005; 11(5): 1910 - 1917. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Hashimoto, S. Asakura, S. Miyake, T. Yamamura, L. Van Kaer, C. Liu, M. Tanimoto, and T. Teshima Stimulation of Host NKT Cells by Synthetic Glycolipid Regulates Acute Graft-versus-Host Disease by Inducing Th2 Polarization of Donor T Cells J. Immunol., January 1, 2005; 174(1): 551 - 556. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. V. Parekh, A. K. Singh, M. T. Wilson, D. Olivares-Villagomez, J. S. Bezbradica, H. Inazawa, H. Ehara, T. Sakai, I. Serizawa, L. Wu, et al. Quantitative and Qualitative Differences in the In Vivo Response of NKT Cells to Distinct {alpha}- and {beta}-Anomeric Glycolipids J. Immunol., September 15, 2004; 173(6): 3693 - 3706. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. S. Mehta, B. Gu, B. Conyers, S. Ouzounov, L. Wang, R. M. Moriarty, R. A. Dwek, and T. M. Block {alpha}-Galactosylceramide and Novel Synthetic Glycolipids Directly Induce the Innate Host Defense Pathway and Have Direct Activity against Hepatitis B and C Viruses Antimicrob. Agents Chemother., June 1, 2004; 48(6): 2085 - 2090. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. S. Metelitsa, H.-W. Wu, H. Wang, Y. Yang, Z. Warsi, S. Asgharzadeh, S. Groshen, S. B. Wilson, and R. C. Seeger Natural Killer T Cells Infiltrate Neuroblastomas Expressing the Chemokine CCL2 J. Exp. Med., May 3, 2004; 199(9): 1213 - 1221. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Nieda, M. Okai, A. Tazbirkova, H. Lin, A. Yamaura, K. Ide, R. Abraham, T. Juji, D. J. Macfarlane, and A. J. Nicol Therapeutic activation of V{alpha}24+V{beta}11+ NKT cells in human subjects results in highly coordinated secondary activation of acquired and innate immunity Blood, January 15, 2004; 103(2): 383 - 389. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Schmieg, G. Yang, R. W. Franck, and M. Tsuji Superior Protection against Malaria and Melanoma Metastases by a C-glycoside Analogue of the Natural Killer T Cell Ligand {alpha}-Galactosylceramide J. Exp. Med., December 1, 2003; 198(11): 1631 - 1641. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. T. Wilson, C. Johansson, D. Olivares-Villagomez, A. K. Singh, A. K. Stanic, C.-R. Wang, S. Joyce, M. J. Wick, and L. Van Kaer The response of natural killer T cells to glycolipid antigens is characterized by surface receptor down-modulation and expansion PNAS, September 16, 2003; 100(19): 10913 - 10918. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Hayakawa, S. Rovero, G. Forni, and M. J. Smyth {alpha}-Galactosylceramide (KRN7000) suppression of chemical- and oncogene-dependent carcinogenesis PNAS, August 5, 2003; 100(16): 9464 - 9469. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.-i. Fujii, K. Shimizu, C. Smith, L. Bonifaz, and R. M. Steinman Activation of Natural Killer T Cells by {alpha}-Galactosylceramide Rapidly Induces the Full Maturation of Dendritic Cells In Vivo and Thereby Acts as an Adjuvant for Combined CD4 and CD8 T Cell Immunity to a Coadministered Protein J. Exp. Med., July 21, 2003; 198(2): 267 - 279. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. J. J. van der Vliet, J. W. Molling, N. Nishi, A. J. Masterson, W. Kolgen, S. A. Porcelli, A. J. M. van den Eertwegh, B. M. E. von Blomberg, H. M. Pinedo, G. Giaccone, et al. Polarization of V{alpha}24+ V{beta}11+ Natural Killer T Cells of Healthy Volunteers and Cancer Patients Using {alpha}-Galactosylceramide-loaded and Environmentally Instructed Dendritic Cells Cancer Res., July 15, 2003; 63(14): 4101 - 4106. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| Cancer Research | Clinical Cancer Research |
| Cancer Epidemiology Biomarkers & Prevention | Molecular Cancer Therapeutics |
| Molecular Cancer Research | Cancer Prevention Research |
| Cancer Prevention Journals Portal | Cancer Reviews Online |
| Annual Meeting Education Book | Meeting Abstracts Online |
, patient 3; 
, patient 5; •, patient 9; 
, patient 17).
