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IMMUNOBIOLOGY
From the Myeloma and Transplantation Research Center,
University of Arkansas for Medical Sciences, Little Rock, AR.
Multiple myeloma (MM) is a B-cell malignancy. The monoclonal
immunoglobulin, secreted by myeloma plasma cells, carries unique antigenic determinants (idiotype [Id]) that can be regarded as a
tumor-specific antigen. Id-based immunotherapy has been explored in
myeloma patients for the purpose of enhancing or inducing Id-specific immune responses that might lead to tumor destruction. However, despite
some evidence obtained from mouse plasmacytoma models, it is still
unclear whether Id-specific immunity may play a role in the regulation
of tumor cells in MM. In the current study, using dendritic cells (DCs)
as antigen-presenting cells, autologous Id-specific cytotoxic T
lymphocyte (CTL) lines containing both CD4+ and
CD8+ T cells were generated from myeloma patients.
The results show that Id-specific CTLs not only recognized and
lysed autologous Id-pulsed DCs but also significantly killed the
autologous primary myeloma cells. The cytotoxicity against the primary
tumor cells was major histocompatibility complex (MHC) class I- and,
to a lesser extent, class II-restricted, indicating that myeloma cells could process Id protein and present Id peptides in the context of
their surface MHC molecules. Furthermore, the CTLs lysed the target cells mainly through the perforin-mediated pathway because Concanamycin A, but not Brefeldin A Multiple myeloma (MM) is a B-cell malignancy that
accounts for 1% of all cancers and 10% of all hematologic
malignancies. With conventional chemotherapy, complete remission rates
have not exceeded 5%, and median survival has not been extended beyond 3 years.1,2 High-dose chemotherapy followed by stem cell support results in long-term, disease-free survival in one third of
myeloma patients.3 However, relapse of the underlying
disease remains the primary cause of treatment failure.4
Therefore, novel alternative therapeutic strategies aiming at improved
outcome in these patients are much needed.
Tumor vaccines based either on immunization against specific tumor
antigens or on adoptive transfer of ex vivo-generated lymphocytes, with
high levels of specific reactivity against tumor antigens, represent attractive approaches.5-8 In MM, the idiotype
(Id) determinant of the paraprotein can be regarded as a tumor-specific marker and has been used for immunotherapy of MM.9-14
Although modest and transient, Id-specific immune responses can be
induced in 25% to 100% of these patients. However, clinical
responses, defined by a significant reduction in M components, have
been observed in few patients after Id vaccination.11,13
Thus, the role of Id-specific immune responses in the regulation of
tumor cell growth is poorly understood, and, more important, whether Id-specific cytotoxic T lymphocytes (CTLs) lyse the primary myeloma cells has not been demonstrated in MM. It is still unclear whether the
induced/enhanced Id-specific immune response after immunization is relevant to or responsible for the clinical response observed in patients.
The objectives of this study were to generate Id-specific CTLs from
myeloma patients and to examine the efficacy of these T cells in lysis
of their target cells, including Id-pulsed dendritic cells (DCs) and
autologous primary myeloma cells. Id-specific CTLs were generated by
using autologous adherent peripheral blood mononuclear cell-derived DCs
as antigen-presenting cells (APCs). We show for the first time that
Id-pulsed DCs reproducibly led to successful in vitro generation of
Id-specific CTLs capable of killing the autologous primary myeloma cells.
Isolation of idiotype protein
Isolation of primary myeloma plasma cells
Generation of dendritic cells Peripheral blood mononuclear cells (PBMCs) from the same 2 patients were isolated from leukapheresis-collected blood cells using Ficoll-Hypaque (Amersham Pharmacia Biotech AB) gradient centrifugation. Aliquots of PBMCs were cryopreserved in 80°C until use. The cryopreserved PBMCs were recovered and resuspended at
5 × 106 cells/mL in RPMI 1640 supplemented with 10%
fetal calf serum, 1 mM glutamine, 100 U/mL penicillin, and 100 µg/mL
streptomycin (referred to as complete medium) (Gibco, Grand Island,
NY). PBMCs were incubated in a humidified incubator at 37°C in 5%
CO2 for 2 hours to allow cell adhesion. Nonadherent cells
were removed by gentle washes. The adherent cells were then cultured in
complete medium supplemented with GM-CSF (100 ng/mL) and IL-4 (100 ng/mL; both from Schering-Plough Research Institute, Kenilworth, NJ) for 7 days, with further addition of cytokines on days 3 and 5. During
the culture, the Id or an allogeneic paraprotein (both at 50 µg/mL)
was added on days 1 and 5 to pulse the cells. After 7 days of culture,
cells were harvested and phenotypically characterized to ensure they
met the typical phenotype of immature DCs that is, CD3 ,
CD4, CD14, CD19,
CD83, CD1a+, CD40+,
CD80+, CD86+, HLA-DR+,
HLA-ABC+.15,16
Generation of idiotype-specific cytotoxic C lymphocytes CTLs were elicited from autologous PBMCs using repeated stimulation by DCs pulsed with myeloma Id. Washed PBMCs (2 × 106/mL) were resuspended in complete medium containing recombinant IL-2 (30 IU/mL; Chiron, Emeryville, CA) and IL-7 (5 ng/mL; R&D Systems, Minneapolis, MN), and Id-pulsed autologous DCs (2-5 × 105/mL) were added to primary cultures. The cells were incubated in 50-mL tissue culture flasks at 37°C in 5% CO2/95% air. Responder cells were restimulated with Id-pulsed autologous DCs every 2 weeks, and the cultures were fed every 5 days with fresh medium containing recombinant IL-2 and IL-7. After 3 to 4 cycles of antigen stimulation and selection, T-cell lines were established and cells were expanded in complete medium containing rIL-2 (30 IU/mL) and IL-7 (5 ng/mL) for 2 weeks and then subjected to functional tests.Proliferation assays T cells (5 × 104/100 µL/well) were seeded into 96-well flat-bottom tissue culture plates. DCs pulsed with the autologous Id, the allogeneic paraprotein from the other patient, or an isotype-matched Id protein from other patients were added to cultures in different concentrations, and cultures were incubated at 37°C in 5% CO2/95% air for 4 days. T-cell proliferation was measured using overnight incubation with [3H]-thymidine (0.5 µCi/well; Amersham Pharmacia Biotech, Piscataway, NJ). Results from triplicate cultures are given as the arithmetic means.Cytotoxicity assays Target cells included autologous DCs pulsed with the autologous Id, the allogeneic paraprotein from the other patient and an isotype-matched Id protein, thawed autologous or allogeneic primary myeloma cells from the other patient, and Ficoll-Hypaque centrifuged to get rid of dead cells. Cells were labeled with sodium chromium Cr 51 chromate for 1 hour, and 1 × 104 target cells/well were mixed with various numbers of effector cells in a standard 4-hour cytotoxicity assay using 96-well round-bottom plates. All assays were performed at least in triplicate. Results are shown as mean percentage 51Cr release calculated as follows: [(sample counts spontaneous counts)/(maximum counts spontaneous counts)]
×100. Spontaneous release was less than 25% of the maximum
[51Cr] uptake.
To determine whether the cytolytic activity was restricted by major histocompatibility complex (MHC) class I or II molecules, antibodies against human leukocyte antigen (HLA)-ABC (W6/32; Serotec, Oxford, United Kingdom) or HLA-DR (B8.12.2; Immunotech, Marseilles, France) and an isotopic control mouse IgG (Immunotech) at 10 µg/mL were added to the cultures at the initiation of the assay. Inhibition of perforin- and Fas-mediated pathways of cytolysis Concanamycin A (CMA; Sigma), an inhibitor of vacuolar type H+-adenosine triphosphate (ATPase), or Brefeldin A (Sigma) was used as a selective inhibitor of perforin-mediated and Fas-mediated cytotoxicity,17 respectively. Effector cells were pretreated with 100 nmol/L CMA or 10 µmol/L Brefeldin A for 2 hours and assayed for cytotoxicity in the presence of the drugs.Enzyme-linked immunospot assay The detailed methods of the enzyme-linked immunospot (ELISPOT) assay for the enumeration of antigen-specific, cytokine-secreting cells have been described previously.18,19 Briefly, plates (Millititer-HAM; Millipore, Bedford, MA) were coated with mouse monoclonal anti-human interferon (IFN-) (1 µg/mL), IL-4 (2.5 µg/mL), or tumor
necrosis factor (TNF)- (2.5.µg/mL) (R&D Systems). Cultured T cells
(104 cells/well) were added and incubated with Id-pulsed
DCs for 36 hours at 37°C. Cells incubated without target cells or
with phytohemagglutinin (10 µg/mL; Sigma) were used as controls.
After incubation, cells were detached from the plates by washing, and
polyclonal anti-human IFN- (250 ng/mL), IL-4 (1 µg/mL) (R&D
System), or TNF- (4 µg/mL; Serotec) was added and incubated for 2 hours. The spots were developed by sequential incubation with
biotinylated antigoat (1/500; IFN- and IL-4) or antirabbit (1/100;
TNF-) antibodies (Vector Labs, Burlingame, CA), followed by
peroxidase staining, using the substrate 3-amino-9-ethyl-carbazol
(Sigma). Spots corresponding to the cytokine-secreting cells were
enumerated under a dissection microscope (Stemi SV6; Zeiss, Jena,
Germany). All samples were run in duplicate. Data are expressed as the
mean number of cytokine-secreting cells/104 T cells.
Immunophenotyping Immunophenotyping was carried out on all cultured cells using a fluorescence-activated cell scan (Facscan; Becton Dickinson, San Jose, CA) and analyzed by Lysis II program. Briefly, cells were first washed twice in washing buffer (phosphate-buffered saline + 2% bovine serum albumin and 0.5% sodium azide). Phycoerythrin- or fluorescein isothiocyanate-conjugated monoclonal antibodies (Immunotech) at the manufacturer's recommended concentrations were added to each cell pellet, and the cells were incubated on ice for 30 minutes before they were washed 3 times and resuspended in phosphate-buffered saline ready for analysis. Controls consisted of cells stained with irrelevant mouse IgG antibodies.
Idiotype-pulsed dendritic cells induce idiotype-specific T-cell responses in vitro To enhance the immunogenicity of Id, we used DCs as APCs. DCs were successfully generated from adherent cells of PBMCs after 7 days of culture and confirmed by immunophenotyping showing that the cells were CD3, CD4, CD14,
CD19, CD83, CD1a+,
CD40+, CD80+, CD86+,
HLA-DR+, and HLA-ABC+ (Figure
1).
We initially examined the immune response to myeloma Id in primary
cultures. No significant proliferative response was observed in any of
the repeated experiments using either freshly recovered PBMC or rIL-2
(50 IU/mL) pretreated PBMCs (data not shown). However, after repeated
rounds of T-cell stimulation with Id-pulsed DCs, Id-specific T-cell
lines were generated and propagated from PBMCs from both myeloma
patients. Id-specific proliferative responses were observed when T
cells were rechallenged with the autologous Id-pulsed DCs (Figure
2A-B). No proliferative response was
noted when T cells were restimulated with autologous DCs pulsed with the allogeneic or isotype-matched Id proteins. These results suggest that autologous T cells could recognize immune epitope(s) within the
myeloma Id proteins, which had been taken up, processed, and presented
by DCs.
The T-cell proliferative responses observed in both patients were blocked partially by monomorphic monoclonal antibody to HLA class I and II molecules (Figure 2C). In patient 1, the blocking was observed predominantly with the HLA class I antibody, whereas in patient 2, the blocking was observed mainly with the HLA class II antibody. A combination of these 2 antibodies produced a synergistic inhibition of the proliferative response (data not shown). These results indicate that the recognition of Id T-cell epitopes was mediated through MHC class I and II molecules. This was supported by flow cytometry analysis showing that the Id-specific T-cell lines consisted of approximately 45% CD8+ and 55% CD4+ T cells. Cytolytic activity of the T cells against idiotype-pulsed dendritic cells To test whether an Id-specific CTL response could be effectively induced with Id-pulsed DCs, we next assessed the cytotoxic activity of Id-specific T cells in vitro. As target cells, autologous DCs and DCs pulsed with autologous, allogeneic, or isotype-matched Id proteins were used. As shown in Figure 3A, Id-specific CTL lines lysed the autologous DCs pulsed with the autologous Id. No significant killing was observed in autologous DCs without antigen pulsing or in autologous DCs pulsed with the allogeneic or isotype-matched Id proteins (Figure 3B). The results indicate that the T cells were able to mediate Id-specific cytotoxicity.
To determine MHC restriction of these Id-specific CTL lines, we studied the inhibitory effect of anti-HLA antibodies on their cytolysis activity. Blocking MHC class I (HLA-ABC) or class II (HLA-DR) molecules of the target cells resulted in an inhibition of the cytotoxicity of the Id-specific CTL against Id-pulsed DCs (Figure 3C). The result further confirmed that the immune epitopes within myeloma Id were presented in association with MHC class I and class II molecules of the patients. Idiotype-specific cytotoxic T lymphocytes lyse primary myeloma plasma cells in vitro To evaluate whether the Id-specific immune response is relevant to a clinical antimyeloma effect, we examined whether Id-specific CTLs lysed autologous primary myeloma cells. Using the CTL lines generated from the patients as effector cells, significant killing of the autologous myeloma cells was observed (Figure 4A). No killing was noted when the allogeneic primary myeloma cells from the other patient or when the natural killer-sensitive target cells K562 were used as target cells (Figure 4B). Blocking of HLA-ABC and, to a lesser extent, HLA-DR molecules of the target cells inhibited the cytotoxicity of the CTLs (Figure 4C). Thus, the results demonstrate that Id-specific CTLs are capable of mediating MHC-restricted cytotoxicity against the autologous primary myeloma cells and suggest that myeloma cells express, on their surfaces, T-cell epitopes derived from Id proteins in the context of MHC molecules.
To examine whether the same T cells mediated the killing of both the Id-pulsed DCs and primary myeloma cells, a cold target inhibition assay was performed. In these experiments, CTLs were first incubated for 2 hours at 37°C with cold, unlabeled autologous DCs pulsed with the Id or the allogeneic Id proteins or with autologous primary tumor cells. As shown in Figure 4D, preincubation of CTLs with the autologous DCs pulsed with the Id, but not the allogeneic or isotype-matched Id proteins, led to diminished killing of the autologous 51Cr-labeled primary tumor cells. On the other hand, incubation of CTLs with autologous, but not allogeneic, primary tumor cells first resulted in reduced lysis of the autologous Id-pulsed DCs (Figure 4E). These results indicate that the same T cells were responsible for the killing of both target cells. Idiotype-specific cytotoxic T lymphocyte response is mediated by the perforin exocytosis pathway To investigate whether the cytotoxic function of Id-specific CTLs was mediated by the FasL/Fas or perforin systems, we examined the effect of CMA and Brefeldin A, the selected inhibitors of perforin-mediated and Fas-mediated cytotoxicity,17 respectively. Treatment of the CTL lines with Brefeldin A induced less than 20% inhibition in the lytic activity of Id-specific CTLs (Figure 5), indicating that the Fas/FasL system was not the main pathway of cytotoxicity mediated by our Id-specific CTLs. This result was supported by flow cytometry analysis showing low or absent FasL expression by the T cells (data not shown). In contrast, when we studied the inhibition of cytotoxicity by pretreatment with CMA, the cytolytic activity was almost completely abrogated (Figure 5). Treatment of the CTLs with Brefeldin A or CMA did not induce apoptosis in the cells (data not shown). Taken together, these results indicate that the cytotoxic function of Id-specific CTLs was mediated mainly by the perforin-dependent pathway.
Cytokine production To characterize cytokine-secretion profile and its corresponding subsets of T cells, an ELISPOT assay was used to detect cytokine secretion. As exemplified by the experiments depicted in Figure 6A-B, a small number of IFN- secreting
cells was detected in the unstimulated T cells. After rechallenge with
Id-pulsed autologous DCs, elevated numbers of IFN-- and
TNF--secreting cells were demonstrated. Levels of IL-4 secreting
cells were extremely low or undetectable. Incubation with DCs only, or
with DCs pulsed with the allogeneic Id protein from the other patient,
did not significantly affect the number of cytokine-secreting cells.
These results were reproduced in the repeated experiments and indicate that the effector cells were of the type-1 T
cells.20,21
In this study, by using Id-pulsed DCs as APCs, we have generated autologous Id-specific CTL lines from patients with MM. These Id-specific CTLs were able to recognize and lyse not only autologous Id-pulsed DCs but also autologous primary myeloma plasma cells, justifying the exploration of Id-based immunotherapy in MM. Results observed in this study indicate that myeloma Id contains T-cell
epitopes recognized by cells mediating proliferative and cytotoxic
responses in an MHC-restricted manner. Both the proliferative and the
cytotoxic responses of the CTLs generated from 2 patients were
restricted by MHC class I and class II molecules, and the CTLs
contained CD4+ and CD8+ T cells. The
observation is in agreement with some previous studies showing that the
Id produced by malignant B cells contains tumor-specific protein
sequences that stimulated CD4+ and CD8+ T
cells.23-29 Furthermore, the cytokine release by the CTLs
was analyzed by the ELISPOT assay. The production of a significant amount of IFN- Plasma cells represent the terminal differentiation stage of B-cell development. In MM, plasma cells constitute at least 10%, and can even reach more than 90%, of the total bone marrow cell count.22 One important aspect is whether the myeloma plasma cells can be recognized and regulated by tumor-specific T cells. Our previous study has shown that freshly isolated primary myeloma cells can activate T cells; they stimulate alloreactive T cells and present recall antigens to autologous T cells,30 suggesting that myeloma cells can act as APCs. The results of the current study demonstrate that Id-specific CTLs lyse the autologous plasma cells in an MHC-restricted manner, indicating that the tumor cells can process Id protein and present Id peptides in the context of MHC molecules on the cell surfaces. Taken together, these findings indicate that myeloma plasma cells express tumor antigens on their surfaces and are susceptible to tumor-specific, T cell-mediated lysis. Because we used CTL lines in the current study that might have contained T cells with different specificity, it was important to examine whether the same T cells lysed both Id-pulsed DCs and primary tumor cells. As shown by the results of cold target inhibition experiments in Figure 4, it is clear that preincubation of the CTLs with Id-pulsed DCs inhibited the killing of primary tumor cells and vice versa. Thus, we can conclude that the same T cells mediated the killing. This is an important issue because it indicates that by using the same Id proteins to vaccinate patients, the generation of a strong CTL response against the Id proteins could lead to tumor destruction. Our results also demonstrate that the immune response may be directly against the idiotypic determinants because the T cells responded only to the autologous Id protein, not to the allogeneic or isotype-matched Id proteins. Furthermore, we and others11,28,29 have shown that T-cell responses induced by Id proteins also were against synthetic peptides corresponding to the complementarity-determining region I-III of the Id proteins. CTL recognition of target cells through their T-cell receptors activates 2 distinct mechanisms of cell lysis.31,32 The first is granule exocytosis mediated by the pore-forming perforin and granzyme A and B. The second involves interaction between the Fas ligand on effector cells and Fas molecules expressed on the target cells. In the current study, the cells appeared to lyse the target cells mainly through the perforin-mediated pathway because CMA, the selective blocking agent, induced 90% inhibition of cytotoxicity. Brefeldin A, a selective inhibitor of the Fas-mediated pathway, resulted in less than 20% inhibition. Hence, these findings are in agreement with our previous study showing that myeloma-reactive allospecific T cells also lysed target cells through the perforin pathway33 and are of special importance in view of published results on Fas expression on myeloma cells. Landowski et al34 have shown that Fas antigen point mutation was detected in 10% of patients' bone marrow samples. The mutations were located in the cytoplasmic region involved in the transduction of an apoptotic signal and, thus, render the cells resistant to Fas-induced apoptosis. Furthermore, they found that myeloma cells induced to be drug resistant also became resistant to Fas-mediated apoptosis.35 Thus, the use of CTLs that are cytotoxic through the Fas-mediated pathway may be limited, but the pore-forming CTLs can be used for the treatment of drug-resistant myeloma. In conclusion, our results demonstrate that by using DCs as APCs, autologous Id-specific CTLs can be generated from myeloma patients. These T cells are efficient in killing Id-pulsed autologous DCs and primary myeloma cells from patients. Thus, our study provides strong and direct evidence to support the application of Id-based immunotherapies in MM.
Submitted August 9, 2000; accepted October 31, 2000.
Supported in part by a grant from the Leukemia and Lymphoma Society. Q.Y. is a recipient of the Leukemia and Lymphoma Society Translational Research Award.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Qing Yi, Myeloma and Transplantation Research Center, University of Arkansas for Medical Sciences, 4301 W Markham St, Slot 776, Little Rock, AR 72205; e-mail: yiqing{at}exchange.uams.edu.
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L. Hansson, A. O. Abdalla, A. Moshfegh, A. Choudhury, H. Rabbani, B. Nilsson, A. Osterborg, and H. Mellstedt Long-term Idiotype Vaccination Combined with Interleukin-12 (IL-12), or IL-12 and Granulocyte Macrophage Colony-Stimulating Factor, in Early-Stage Multiple Myeloma Patients Clin. Cancer Res., March 1, 2007; 13(5): 1503 - 1510. [Abstract] [Full Text] [PDF]
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J. Qian, S. Wang, J. Yang, J. Xie, P. Lin, M. E. Freeman III, and Q. Yi Targeting Heat Shock Proteins for Immunotherapy in Multiple Myeloma: Generation of Myeloma-Specific CTLs Using Dendritic Cells Pulsed with Tumor-Derived gp96 Clin. Cancer Res., December 15, 2005; 11(24): 8808 - 8815. [Abstract] [Full Text] [PDF]
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R. B. Batchu, A. M. Moreno, S. M. Szmania, G. Bennett, G. C. Spagnoli, S. Ponnazhagan, B. Barlogie, G. Tricot, and F. van Rhee Protein Transduction of Dendritic Cells for NY-ESO-1-Based Immunotherapy of Myeloma Cancer Res., November 1, 2005; 65(21): 10041 - 10049. [Abstract] [Full Text] [PDF]
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K. Kawano, S. Ferrone, and C. G. Ioannides Functional Idiotopes: Tumor Antigen-Directed Expression of CD8+ T-Cell Epitopes Nested in Unique NH2-terminal VH Sequence of Antiidiotypic Antibodies? Cancer Res., July 15, 2005; 65(14): 6001 - 6004. [Abstract] [Full Text] [PDF]
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S. B. Rew, K. Peggs, I. Sanjuan, A. R. Pizzey, Y. Koishihara, S. Kawai, M. Kosaka, S. Ozaki, B. Chain, and K. L. Yong Generation of Potent Antitumor CTL from Patients with Multiple Myeloma Directed against HM1.24 Clin. Cancer Res., May 1, 2005; 11(9): 3377 - 3384. [Abstract] [Full Text] [PDF]
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C. Choi, M. Witzens, M. Bucur, M. Feuerer, N. Sommerfeldt, A. Trojan, A. Ho, V. Schirrmacher, H. Goldschmidt, and P. Beckhove Enrichment of functional CD8 memory T cells specific for MUC1 in bone marrow of patients with multiple myeloma Blood, March 1, 2005; 105(5): 2132 - 2134. [Abstract] [Full Text] [PDF]
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Q. Yi and L. W. Kwak Monocyte-Derived Dendritic Cells: A Promising Armament for Immunotherapy in Human Malignancies Clin. Cancer Res., February 1, 2005; 11(3): 966 - 967. [Full Text] [PDF]
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J. L. Murray, M. Gillogly, K. Kawano, C. L. Efferson, J. E. Lee, M. Ross, X. Wang, S. Ferrone, and C. G. Ioannides Fine Specificity of High Molecular Weight-Melanoma-Associated Antigen-Specific Cytotoxic T Lymphocytes Elicited by Anti-Idiotypic Monoclonal Antibodies in Patients with Melanoma Cancer Res., August 1, 2004; 64(15): 5481 - 5488. [Abstract] [Full Text] [PDF]
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B. Barlogie, J. Shaughnessy, G. Tricot, J. Jacobson, M. Zangari, E. Anaissie, R. Walker, and J. Crowley Treatment of multiple myeloma Blood, January 1, 2004; 103(1): 20 - 32. [Abstract] [Full Text] [PDF]
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L. Hansson, H. Rabbani, J. Fagerberg, A. Osterborg, and H. Mellstedt T-cell epitopes within the complementarity-determining and framework regions of the tumor-derived immunoglobulin heavy chain in multiple myeloma Blood, June 15, 2003; 101(12): 4930 - 4936. [Abstract] [Full Text] [PDF]
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T. Rasmussen, L. Hansson, A. Osterborg, H. E. Johnsen, and H. Mellstedt Idiotype vaccination in multiple myeloma induced a reduction of circulating clonal tumor B cells Blood, June 1, 2003; 101(11): 4607 - 4610. [Abstract] [Full Text] [PDF]
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E. Orsini, R. Bellucci, E. P. Alyea, R. Schlossman, C. Canning, S. McLaughlin, P. Ghia, K. C. Anderson, and J. Ritz Expansion of Tumor-specific CD8+ T Cell Clones in Patients with Relapsed Myeloma after Donor Lymphocyte Infusion Cancer Res., May 15, 2003; 63(10): 2561 - 2568. [Abstract] [Full Text] [PDF]
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C. Milazzo, V. L. Reichardt, M. R. Muller, F. Grunebach, and P. Brossart Induction of myeloma-specific cytotoxic T cells using dendritic cells transfected with tumor-derived RNA Blood, February 1, 2003; 101(3): 977 - 982. [Abstract] [Full Text] [PDF]
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 M. V. Dhodapkar, J. Krasovsky, and K. Olson T cells from the tumor microenvironment of patients with progressive myeloma can generate strong, tumor-specific cytolytic responses to autologous, tumor-loaded dendritic cells PNAS, October 1, 2002; 99(20): 13009 - 13013. [Abstract] [Full Text] [PDF]
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 C. Paludan, K. Bickham, S. Nikiforow, M. L. Tsang, K. Goodman, W. A. Hanekom, J.-F. Fonteneau, S. Stevanovic, and C. Munz Epstein-Barr Nuclear Antigen 1-Specific CD4+ Th1 Cells Kill Burkitt's Lymphoma Cells J. Immunol., August 1, 2002; 169(3): 1593 - 1603. [Abstract] [Full Text] [PDF]
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Y.-J. Wen, R. Min, G. Tricot, B. Barlogie, and Q. Yi Tumor lysate-specific cytotoxic T lymphocytes in multiple myeloma: promising effector cells for immunotherapy Blood, May 1, 2002; 99(9): 3280 - 3285. [Abstract] [Full Text] [PDF]
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A. C. Armstrong, S. Dermime, C. G. Allinson, T. Bhattacharyya, K. Mulryan, K. R. Gonzalez, P. L. Stern, and R. E. Hawkins Immunization with a Recombinant Adenovirus Encoding a Lymphoma Idiotype: Induction of Tumor-Protective Immunity and Identification of an Idiotype-Specific T Cell Epitope J. Immunol., April 15, 2002; 168(8): 3983 - 3991. [Abstract] [Full Text] [PDF]
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 A. Badros, B. Barlogie, E. Siegel, M. Cottler-Fox, M. Zangari, A. Fassas, C. Morris, E. Anaissie, F. Van Rhee, and G. Tricot Improved Outcome of Allogeneic Transplantation in High-Risk Multiple Myeloma Patients After Nonmyeloablative Conditioning J. Clin. Oncol., March 1, 2002; 20(5): 1295 - 1303. [Abstract] [Full Text] [PDF]
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K. Kronenberger, A. Dieckmann, M. Selmayr, J. Strehl, U. Wahl, H. Lindhofer, G. Kraal, and R. Mocikat Impact of the lymphoma idiotype on in vivo tumor protection in a vaccination model based on targeting antigens to antigen-presenting cells Blood, February 15, 2002; 99(4): 1327 - 1331. [Abstract] [Full Text] [PDF]
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S. Harig, M. Witzens, A. M. Krackhardt, A. Trojan, P. Barrett, R. Broderick, A. J. Zauls, and J. G. Gribben Induction of cytotoxic T-cell responses against immunoglobulin V region-derived peptides modified at human leukocyte antigen-A2 binding residues Blood, November 15, 2001; 98(10): 2999 - 3005. [Abstract] [Full Text] [PDF]
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D. M.-Y. Sze, G. Giesajtis, R. D. Brown, M. Raitakari, J. Gibson, J. Ho, A. G. Baxter, B. Fazekas de St Groth, A. Basten, and D. E. Joshua Clonal cytotoxic T cells are expanded in myeloma and reside in the CD8+CD57+CD28- compartment Blood, November 1, 2001; 98(9): 2817 - 2827. [Abstract] [Full Text] [PDF]
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Top
Abstract
Introduction
), the
allogeneic Id from the other patient (
), or an isotype-matched (
)
Id protein. (C) Inhibition by antibodies against MHC class I and class
II or an isotypic control IgG on Id-induced proliferation of CTLs from
patients 1 (
) and 2 (
). These results are representative of 3 independent experiments.
).