|
|||||||||
|
|
|
|
|
|
|
|
|
|
|
IMMUNOBIOLOGY
From the Department of Adult Oncology, Dana-Farber
Cancer Institute, Department of Medicine, Brigham and Women's
Hospital, Harvard Medical School, Boston, MA.
Chronic lymphocytic leukemia (CLL) cells are ineffective
antigen-presenting cells (APCs) although CD40-activated CLL cells can
stimulate proliferation of autologous and allogeneic T cells. We
examined the antigen-presenting capacity of CD40-activated CLL cells as
well as dendritic cells pulsed with apoptotic bodies of CLL cells to
generate autologous and allogeneic immune responses against CLL cells.
Both APC types were capable of generating T-cell lines that proliferate
specifically in response to unstimulated CLL cells. Whereas cytotoxic
responses against stimulated and unstimulated CLL cells could be
repeatedly generated by allogeneic healthy donors, autologous cytotoxic
immune responses against CD40-activated and native CLL cells were
rarely detected. However, T cells isolated from patients with CLL could
recognize and lyse allogeneic stimulated and unstimulated CLL cells,
demonstrating that cytotoxic T cells from these tumor-bearing patients
are functionally intact.
(Blood. 2002;100:167-173) Chronic lymphocytic leukemia (CLL) is an attractive
candidate to examine immunotherapeutic approaches because it is a
slow-growing tumor, allowing time for the generation of an immune
response against the tumor cells. CLL cells express major
histocompatibility complex (MHC) class I and class II molecules and
also express a specific tumor antigen, the idiotype, which has been
shown to be a target of T- and B-cell-mediated immune
responses.1-5 Following allogeneic stem cell
transplantation, CLL cells appear to be good targets of a
graft-versus-leukemia effect.6 However, a number of immune
dysregulations occur in CLL with patients often exhibiting severe
hypogammaglobulinemia, impaired immunoglobulin class switching, and
diminished germinal center formation. This results in an increased susceptibility to bacterial infections that contributes greatly to
morbidity and mortality in CLL.7-9
It is not clear what role T cells play in the observed B-cell defects
and immunosuppression in CLL. A number of T-cell defects have been well
characterized including inversion of the CD4/CD8 ratio,10
enhanced susceptibility of CD4 cells to FAS (CD95) ligand-induced
apoptosis,11 and expression of an oligoclonal T-cell
receptor (TCR) V Despite these T-cell alterations, we have previously demonstrated that
autologous cytotoxic T-cell lines can be generated against
immunoglobulin-derived peptides when these immunogenic peptides are
presented by antigen-presenting cells (APCs) and that once established
these T-cell lines are capable of killing CLL cells.21,22
These data demonstrate that because cytotoxic T cells can be expanded
and are capable of recognizing and killing autologous tumor cells, they
must be present within the T-cell repertoire. Although CLL cells can be
a target of a T-cell-mediated cytotoxic response once these cytotoxic
T-cell lines have been established ex vivo, CLL cells are ineffective
stimulator cells in allogeneic as well as autologous mixed lymphocyte
cultures and it has been extremely difficult to initiate autologous
T-cell responses against unstimulated CLL cells.23 The low
immunogenicity of the CLL cells is due at least in part to their low
levels of expression of costimulatory and adhesion molecules required
for the initiation of an effective immune response. CD40 has been shown
to be an effective tool to up-regulate expression of these molecules on
both normal and malignant B cells including CLL
cells.24-26 Moreover, CD40-activated normal and malignant
B cells induce T-cell responses27-29 and transfection of
CLL cells with replication-defective adenovirus encoding murine CD154
induced immune responses against unmodified leukemia
cells.30,31
Although CD40-activated normal and lymphoma B cells are effective APCs,
CD40-activated CLL cells might still have defects in antigen
presentation and induction of antitumor responses, because the CLL
cells themselves might be involved in dysregulation of autologous T
cells. Dendritic cells (DCs) have been shown to be effective in
inducing antitumor responses in many cancers including B-cell
lymphoma.2,4,32,33 However, the role of DCs to initiate T-cell immune responses has not previously been investigated in patients with CLL. In the present report we used both CD40-activated CLL cells as well as DCs pulsed with apoptotic bodies of CLL cells as
APCs to test their capacity to induce proliferative and cytotoxic autologous and allogeneic T-cell responses against CLL cells. Both APC
types induced specific proliferating T lymphocytes in response to
unstimulated CLL cells that could be partially blocked by antibodies
against MHC class I and class II molecules suggesting that
CLL-associated antigens are presented by both pathways. Cytotoxicity and specific proliferation could be induced against unstimulated allogeneic CLL cells. However, autologous cytotoxic T-cell responses could rarely be generated against naturally presented peptides. In
keeping with the notion that there are T-cell alterations in the
autologous tumor-bearing host, prior to stimulation we observed enhanced expression of the chemokine receptors CXCR4 and reduced expression of CCR2 on CD4+ cells of patients with CLL
compared to healthy donors. However, T cells isolated from patients
with CLL are capable of mounting cytotoxic immune responses against
allogeneic CLL cells. Therefore, although a number of defects have been
demonstrated in the T cells of CLL patients, including defective
granzyme-perforin pathway and reduced cell numbers, CD8+
cytotoxic T cells from CLL appear to be functionally capable of killing
CLL cells. Taken together these data are consistent with the notion
that to induce effective autologous immune responses against CLL cells,
it will likely be necessary to enhance antigen presentation, perhaps to
overcome tolerance mechanisms on both the antigen-presenting and
effector pathways.
Patients and donors
Cell preparation and isolation of different cell types
Cell culture and cytokines Cells were cultured at 37°C in a 5% CO2 humidified atmosphere. To generate DCs, adherent cells were cultured in Iscoves modified Dulbecco medium (IMDM; Life Technologies, Rockville, MD) supplemented with 10% human serum, 50 µg/mL human transferrin (Boehringer Mannheim, Indianapolis, IN), 5 µg/mL human insulin (Sigma Chemical, St Louis, MO), 15 µg/mL gentamicin (Life Technologies), 2.4 mg/mL HEPES (Mediatech-Cellgro, Herndon, VA), and 2.9 mg/mL L-glutamine (Mediatech-Cellgro), and addition of 10 ng/mL recombinant human IL-4 (rhIL-4; R & D Systems, Minneapolis, MN) and 50 ng/mL recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF; Immunex, Seattle, WA) for 5 days. Cells were then loaded with apoptotic bodies of CLL cells in a DC apoptotic cell ratio
of 1:4. Apoptotic bodies were generated by irradiation of freshly
thawed CLL cells with 50 Gy. Apoptotic cell death was assayed by using
annexin V-fluorescein isothiocyanate (FITC) and propidium iodide
staining, according to the manufacturer's instructions. For maturation
of DCs, 1 µg/mL soluble CD40L (Immunex) was added 24 hours before
stimulation of T cells. Prior to T-cell stimulation residual apoptotic
bodies were depleted by rinsing the cell culture plates. DCs were then
harvested using a cell scraper, irradiated with 32 Gy, and used for
T-cell activation in a ratio of 1:5 to 1:10.
Murine NIH3T3 fibroblasts transfected with the human CD40L were used for stimulation of malignant B cells.29 Malignant B cells were cultured on irradiated CD40L-transfected fibroblasts in IMDM supplemented with 10% human serum, 50 µg/mL human transferrin, 5 µg/mL human insulin, 15 µg/mL gentamicin, 2.4 mg/mL HEPES, and 2.9 mg/mL L-glutamine, and addition of 5 ng/mL rhIL-4 for 3 days. Before stimulation of T cells, CD40L-activated CLL cells were irradiated with 32 Gy. Purified CD3+ T cells (2 × 106) from healthy donors or CLL patients were cultured in RPMI with 10% human serum, 50 U/mL penicillin, 50 µg/mL streptomycin, 15 µg/mL gentamicin (Life Technologies), 2.4 mg/mL HEPES, and 2.9 mg/mL L-glutamine. After stimulation with DCs or CD40L-activated CLL cells, the following cytokines were added to T-cell cultures: 1 ng/mL rhIL-7 (Pepro Tech, Rocky Hill, NJ) was added on day 0, rhIL-2 (kind gift from Dr J. Ritz, Dana-Farber Cancer Institute) was added on days 1 (50 U/mL) and 4 (25 U/mL). T cells were restimulated every week with stimulator cells in a ratio ranging from 1:4 to 1:10. Proliferation and cytotoxicity assays were performed 3 days after the fourth stimulation. Immunophenotyping Immunophenotyping was performed by fluorescence-activated cell sorter analysis to verify isolation and purification steps as well as to investigate activation markers and expression of specific receptors on T and B cells. The following mAbs conjugated with FITC or phycoerythrin (PE) were used: mouse IgG control, CD1c, CD3, CD4, CD5, CD8, CD19, CD20, CD21, CD23, CD28, CD54, CD56, CD70, CD80, CD83, CD86, CD95, CD152, CD154, HLA-ABC, HLA-DR, anti- , anti- (Dako,
Carpinteria, CA); CD2 (Coulter, Fullerton, CA); and CD40, CD45RA,
CD45RO, CD58 (BD Pharmingen, Franklin Lakes, NJ). The following
chemokine receptors were investigated using mAbs against CCR2, CCR3,
CCR5, CXCR1, CXCR2, CXCR3, CXCR4, and CXCR5 (R & D Systems). For
investigation of chemokine receptor expression on circulating T cells
freshly purified peripheral blood mononuclear cells from patients with
CLL and healthy donors were analyzed under exactly identical
circumstances. In case of nonconjugated antibodies a secondary staining
was performed with FITC-conjugated goat anti mouse/rat IgG antibody
(Jackson, West Grove, PA).
Mixed lymphocyte reaction Irradiated (32 Gy) unstimulated CLL cells were used as stimulator cells in a 72-hour thymidine incorporation assay. Briefly, 5 × 104 cultured T cells were used in triplicate and stimulated with stimulator cells in a ratio of 1:1 to 1:4 in a 96-well microtiter U-bottom culture plate. After 72 hours, cells were pulsed with 0.5 µCi 3H-thymidine (0.0185 MBq; NEN, Boston, MA), cultured for another 12 hours, and harvested onto glass fiber filters (Wallac, Turku, Finland). 3H-thymidine incorporation was measured using a Wallac Microbeta Plus Scintillation Counter.Blocking experiments Experiments for blocking of proliferation were performed using murine mAbs against MHC class I and class II. W6/32 (kind gift of Dr K. Anderson, Dana-Farber Cancer Institute) was used to block MHC class I and CR3/43 (Dako) to block MHC class II molecules.Cytotoxicity assay A standard chromium (51Cr) release assay was performed.37 Unstimulated CLL cells, CD40L-activated CLL cells, and K562 cells were labeled with 100 µCi 51Cr (3.7 MBq; NEN) and seeded in 96-well U-bottom microtiter plates at a concentration of 2.5 × 103 in triplicates. Effector cells were added in a ratio of 1:3, 1:10, and 1:30 and cocultured for 4 hours at 37°C in a 5% CO2 humidified atmosphere. After 4 hours the supernatants were harvested and the released 51Cr was measured in a -Counter (Wallac). Spontaneous release was determined by incubation of target cells in medium alone and maximum release was determined by resuspending the wells with 2% Triton X-100.
Specific lysis was determined for each individual experiment as
follows: specific lysis (%) = (experimental Cr51
release  spontaneous 51Cr release)/maximum
51Cr release spontaneous 51Cr
release) × 100.
Statistical analysis Differences between experimental groups were analyzed using the Wilcoxon 2-sample test.
Activation and proliferation of autologous and allogeneic T cells with CD40-activated CLL cells or DCs pulsed with apoptotic bodies of CLL cells as APCs Our previous studies have demonstrated that cytotoxic T-cell responses can be generated against immunoglobulin-derived peptides expressed in CLL cells.21,22 However, cytotoxicity induced against autologous CLL cells in this system was weak. Other tumor-associated antigens are likely also to be presented on CLL cells but do not naturally induce effective antitumor responses. Because these antigens are not yet characterized, it is not possible to use DCs to present these antigens. We sought to investigate specific cellular responses against such naturally presented antigens in CLL. Because CD40 activation is an effective mechanism to increase APC function of malignant B cells including CLL cells, we investigated the proliferative potential of allogeneic and autologous T cells stimulated with CD40-activated CLL cells. Proliferation of allogeneic T cells was enhanced compared to autologous T cells in all experiments performed in response to CD40-activated CLL cells (Figure 1A). DCs have been shown to be effective in inducing autologous antitumor responses against malignant cells in several tumor models. We therefore used DCs pulsed with apoptotic bodies of autologous CLL cells as described in "Patients, methods, and materials." After 28 days of culture, T-cell proliferation was equivalent in response to pulsed DCs compared to CD40-activated CLL cells as stimulator cells (Figure 1B). Immunophenotypic investigation of T cells after stimulation with CD40-activated CLL cells showed no significant difference in proliferation of CD4+ T cells after 8 to 22 days in the autologous compared to the allogeneic setting (Figure 2A). In contrast, reduced proliferation of CD8+ T cells was observed in the autologous compared to the allogeneic setting within this period (Figure 2B).
Specific proliferation and cytotoxicity of activated T cells in response to unstimulated CLL cells To determine whether T-cell lines generated after stimulation with CD40-activated CLL cells could proliferate in response to the native, unstimulated CLL cells, T cells were restimulated weekly 4 times with CD40-activated CLL cells. After 4 weekly stimulations with CD40-activated CLL cells, T cells proliferated specifically in response to unstimulated CLL cells in 3 of 5 allogeneic cases tested (data not shown) and in 5 of 7 autologous cases (Figure 3A). T-cell lines generated against DCs pulsed with apoptotic bodies of CLL cells also demonstrated specific proliferation in response to unstimulated CLL cells in 3 of 4 autologous samples tested (data not shown), demonstrating that DCs in CLL patients must be capable of presenting antigens from the apoptotic CLL cells. Therefore, although unstimulated CLL cells cannot initiate a proliferative T-cell response, once these T cells have been generated, we here demonstrate that unstimulated CLL cells can function to induce proliferation of the established T-cell lines. Addition of either anti-MHC class I or anti-MHC class II mAbs partially reduced specific proliferation against the unstimulated CLL cells, suggesting that antigens are presented by both MHC class I and class II pathways (Figure 3B). Although allogeneic cytotoxic T cells that induced specific killing of CD40-activated or unstimulated CLL cells (Figure 4) could be generated in 4 of 6 experiments performed, cytotoxic immune responses against CD40-activated CLL cells could be generated in this manner in only 1 of 9 autologous patient samples tested (data not shown). Comparable results were obtained after initial stimulation with DCs pulsed with autologous CLL apoptotic bodies, because cytotoxic immune responses could be generated only weakly in 1 of 4 different patient samples tested (data not shown). Addition of anti-CD80 mAb resulted in greatly reduced proliferation and cytotoxicity in both the autologous and allogeneic system (P < .01; data not shown), suggesting that the CD86/CD28 interaction could not substitute for CD28/CD80-mediated costimulation in this system. Although addition of anti-CD86 mAb partially reduced proliferation (P < .05) of both autologous and allogeneic T cells, cytotoxicity of the resulting allogeneic T cells against CLL cells was equal or even enhanced in 5 of 5 experiments performed (data not shown). Addition of IL-12 or mAbs against IL-10 or transforming growth factor (TGF-)
did not enhance cytotoxic immune responses in the autologous system
(data not shown).
Chemokine receptor expression on unstimulated and stimulated autologous and allogeneic T cells We hypothesized that there might be differences in the T-cell populations and activation state of autologous compared to allogeneic healthy T cells prior to priming. We saw no association between T-cell number or the CD4 and CD8 immunophenotype and the observed proliferative and cytotoxic responses (data not shown). To further investigate the T-cell populations, we analyzed chemokine receptor expression on CD4+ T cells of CLL patients compared to healthy donors and observed an altered chemokine receptor expression (Figure 5). CD4+ T cells from CLL patients exhibited enhanced expression of CXCR4 (P = .02) and decreased expression of CCR2 (P = .02).
Cytotoxic response after stimulation with pulsed DCs of T cells from patients with CLL against unstimulated and CD40-activated CLL cells Given that we observed alterations in the chemokine receptor expression profile on T cells from patients with CLL compared to healthy donors, we further questioned whether basic defects reside in the T cells of patients with CLL or potentially in immunomodulatory effects of T cells seen in the autologous setting. We therefore examined whether T cells from patients with previously untreated CLL are capable of killing allogeneic CLL cells. Three patients with CLL underwent leukopheresis and from these preparations T cells were stimulated with DCs pulsed with apoptotic bodies of allogeneic CLL cells (Table 2). The resulting T-cell lines were then examined for their ability to kill the CD40-activated or unstimulated CLL target cells of the same patient from whom the apoptotic CLL cell bodies were generated. Under these circumstances, T cells from patients with CLL induced specific cytotoxicity in all 3 experiments performed using CD40-activated CLL cells as targets and killing of unstimulated CLL cells was induced in 2 of the 3 cases examined (Table 2). Again there was enhanced killing of CD40-activated CLL cells compared to unmodified CLL cells. These data suggest that T cells from patients with CLL are capable of killing allogeneic CLL cells and further that DCs from patients with CLL are capable of providing sufficient antigen presentation to initiate this response. They further demonstrate that T cells from patients with CLL are not globally defective in their cytotoxic function and that the generalized defects in granzyme-perforin or death ligand pathways are unlikely to be solely responsible for the observed absence of autologous cytotoxicity. However, these experiments do not rule out the possibility that in the autologous setting tolerance may occur that is specific for naturally presented peptides expressed on the autologous tumor cells.
The aim of this study was to investigate further the nature of immune responses of autologous and allogeneic T cells against CLL cells. Autologous cytotoxic T cells can be generated against immunoglobulin-derived peptides presented by APCs. The resulting T-cell lines kill unmodified CLL cells, demonstrating that CLL cells must be capable of presenting this tumor antigen. To examine whether T cells could be primed against naturally processed and presented tumor antigens by CD40-activated CLL cells and by DCs pulsed with apoptotic bodies of CLL cells, autologous and allogeneic T cells were stimulated with CD40-activated CLL cells and proliferative and cytotoxic responses against CD40-activated and unmodified CLL cells were examined. Whereas there was no difference in proliferation of CD4+ autologous and allogeneic T cells, we observed reduced proliferation of autologous CD8+ T cells. In most cases we were able to generate T-cell lines that exhibited specific proliferation in response to unmodified CLL cells, an observation that has not previously been reported. Proliferation of the autologous T-cell lines against unmodified CLL cells could be partially blocked by either MHC class I or class II mAbs, suggesting that both class I- and class II-restricted CLL-associated antigens are being presented and recognized by autologous T cells. Cytotoxic immune responses could be generated against autologous CD40-activated and unmodified CLL cells in only 2 patients, suggesting that these specifically proliferating autologous T cells do not provide sufficient T-cell help for induction of cytotoxic T-cell responses. Autologous DCs pulsed with apoptotic bodies of allogeneic CLL cells can present allogeneic CLL cell peptides and induced cytotoxic T-cell responses, suggesting that T cells of patients with CLL are functionally able to generate cytotoxic immune responses against allogeneic CLL cells. The low autologous cytotoxic responses might also be due to primary or secondary alterations in the effector populations, in keeping with differences in chemokine receptor expression on T cells from patients with CLL and healthy donors. Therefore, the interaction in vivo between autologous CLL cells, APCs, and T cells might favor induction of tolerance rather than induction of active antitumor responses. Several reasons might be responsible for the low capacity to induce cytotoxic T-cell responses against autologous CLL cells. CLL cells fail to express or express weakly, critical adhesion and costimulatory molecules and CD154 has been shown to be an effective tool to up-regulate costimulatory molecules on the surface of CLL cells.24-26 We have previously demonstrated that killing of CLL cells by cytotoxic T cells generated against immunoglobulin-derived peptides presented by APCs was increased by either addition of exogenous peptide or by CD40 activation of the CLL target cells.21 This suggests that inadequate antigen presentation as well as costimulation contribute to the low levels of cytotoxicity observed against native CLL cells. In the present study, cytotoxic immune responses against CD40-activated CLL cells were always higher than against unmodified CLL cells. Addition of blocking mAbs against CD80 and CD86 reduced proliferative and cytotoxic immune responses, further demonstrating that adequate costimulation enhances T-cell effector function as well as being necessary for induction of T-cell-mediated immune responses. However, in contrast to other B-cell malignancies, enhancement of costimulation by CD40 activation or by presentation of whole tumor cells by DCs does not appear to be sufficient to induce autologous cytotoxic T-cell responses in CLL and additional factors probably play a role in regulating this response. Tolerance induced by low differences for
self-/non-self-discrimination, deletion of potentially self-reactive T
cells due to thymic expression of the self-protein, or induction of
peripheral T-cell tolerance in the tumor-bearing host may be
responsible for T-cell unresponsiveness in CLL and the difficulty of
initiating antitumor responses against naturally presented antigens.
The superiority of peptide-pulsed APCs in inducing cytotoxic responses compared to CD40-activated CLL cells or pulsed DCs could be explained by the fact that antigen-derived peptides with low natural
immunogenicity might represent subdominant epitopes normally ignored in
the presence of tolerogenic dominant epitopes naturally presented by
CD40-activated CLL cells and pulsed CCs.38 Dominant
epitopes presented by autologous APCs might activate CD4
immunoregulatory cells responsible for induction of tolerance. Thus,
peptides naturally presented by autologous APCs in vivo might not be
ideal to induce autologous cytotoxic immune responses.39
Alteration of the peripheral T-cell pool possibly induced in vivo by
CLL cells might be involved in the reduced capability to induce
cytotoxic T-cell responses against autologous CLL cells. We observed
significantly increased CXCR4 expression and decreased CCR2 expression
on T cells in patients with CLL compared to healthy donors,
suggesting alterations in the circulating T-cell pool in patients with
CLL. CXCR4, which has significantly enhanced expression on
peripheral T cells of patients with CLL compared to healthy donors, is
highly expressed on naïve T cells compared to memory
cells.40 Peripheral cytokines seem to regulate its
expression as IL-2 and TGF In conclusion, these data further demonstrate that specific allogeneic and autologous proliferative T-cell responses can be generated against CLL cells. However, the role of these specific proliferating T cells after stimulation with autologous APCs is not clear and does not result in enhancement of cytotoxicity. These results need to be considered in the development of novel immunotherapeutic strategies in this disease. Future studies are needed to identify and further stimulate the antitumor effector cells and to prevent or reverse induction of tolerance.
We thank David Zahrieh and Donna Neuberg for statistical analysis and Peter Varney for assistance.
Submitted March 14, 2001; accepted February 21, 2002.
Supported by a grant to J.G.G. from the National Cancer Institute (CA 81534) and by the Grayce B. Kerr Foundation. A.M.K. and M.W. were supported by grants from the Deutsche Krebshilfe.
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: John G. Gribben, Dana-Farber Cancer Institute, 44 Binney St, Boston, MA 02115; e-mail: john_gribben{at}dfci.harvard.edu.
1. George AJ, Folkard SG, Hamblin TJ, Stevenson FK. Idiotypic vaccination as a treatment for a B cell lymphoma. J Immunol. 1988;141:2168-2174[Abstract]. 2. Kwak LW, Campbell MJ, Czerwinski DK, Hart S, Miller RA, Levy R. Induction of immune responses in patients with B-cell lymphoma against the surface-immunoglobulin idiotype expressed by their tumors. N Engl J Med. 1992;327:1209-1215[Abstract]. 3. Tao MH, Levy R. Idiotype/granulocyte-macrophage colony-stimulating factor fusion protein as a vaccine for B-cell lymphoma. Nature. 1993;362:755-758[CrossRef][Medline] [Order article via Infotrieve]. 4. Hsu FJ, Benike C, Fagnoni F, et al. Vaccination of patients with B-cell lymphoma using autologous antigen-pulsed dendritic cells. Nat Med. 1996;2:52-58[CrossRef][Medline] [Order article via Infotrieve]. 5. Bendandi M, Gocke CD, Kobrin CB, et al. Complete molecular remissions induced by patient-specific vaccination plus granulocyte-monocyte colony-stimulating factor against lymphoma. Nat Med. 1999;5:1171-1177[CrossRef][Medline] [Order article via Infotrieve]. 6. Khouri IF, Keating M, Korbling M, et al. Transplant-lite: induction of graft-versus-malignancy using fludarabine-based nonablative chemotherapy and allogeneic blood progenitor-cell transplantation as treatment for lymphoid malignancies. J Clin Oncol. 1998;16:2817-2824[Abstract].
7.
Rozman C, Montserrat E.
Chronic lymphocytic leukemia [published erratum appears in N Engl J Med 1995 Nov 30;333(22):1515].
N Engl J Med.
1995;333:1052-1057 8. Keating MJ. Chronic lymphocytic leukemia. Semin Oncol. 1999;26:107-114[Medline] [Order article via Infotrieve]. 9. Cheson BD. Chronic lymphocytic leukemia and hairy-cell leukemia. Curr Opin Hematol. 1994;1:268-277[Medline] [Order article via Infotrieve].
10.
Foa R, Giovarelli M, Jemma C, et al.
Interleukin 2 (IL 2) and interferon-gamma production by T lymphocytes from patients with B-chronic lymphocytic leukemia: evidence that normally released IL 2 is absorbed by the neoplastic B cell population.
Blood.
1985;66:614-619
11.
Tinhofer I, Marschitz I, Kos M, et al.
Differential sensitivity of CD4+ and CD8+ T lymphocytes to the killing efficacy of Fas (Apo-1/CD95) ligand+ tumor cells in B chronic lymphocytic leukemia.
Blood.
1998;91:4273-4281 12. Farace F, Orlanducci F, Dietrich PY, et al. T cell repertoire in patients with B chronic lymphocytic leukemia. Evidence for multiple in vivo T cell clonal expansions. J Immunol. 1994;153:4281-4290[Abstract].
13.
Rezvany MR, Jeddi-Tehrani M, Osterborg A, Kimby E, Wigzell H, Mellstedt H.
Oligoclonal TCRBV gene usage in B-cell chronic lymphocytic leukemia: major perturbations are preferentially seen within the CD4 T-cell subset.
Blood.
1999;94:1063-1069 14. Goolsby CL, Kuchnio M, Finn WG, Peterson L. Expansions of clonal and oligoclonal T cells in B-cell chronic lymphocytic leukemia are primarily restricted to the CD3(+)CD8(+) T-cell population. Cytometry. 2000;42:188-195[CrossRef][Medline] [Order article via Infotrieve].
15.
Totterman TH, Carlsson M, Simonsson B, Bengtsson M, Nilsson K.
T-cell activation and subset patterns are altered in B-CLL and correlate with the stage of the disease.
Blood.
1989;74:786-792 16. Cantwell M, Hua T, Pappas J, Kipps TJ. Acquired CD40-ligand deficiency in chronic lymphocytic leukemia. Nat Med. 1997;3:984-989[CrossRef][Medline] [Order article via Infotrieve]. 17. Rossi E, Matutes E, Morilla R, Owusu-Ankomah K, Heffernan AM, Catovsky D. Zeta chain and CD28 are poorly expressed on T lymphocytes from chronic lymphocytic leukemia. Leukemia. 1996;0:494-497. 18. Dazzi F, D'Andrea E, Biasi G, et al. Failure of B cells of chronic lymphocytic leukemia in presenting soluble and alloantigens. Clin Immunol Immunopathol. 1995;75:26-32[CrossRef][Medline] [Order article via Infotrieve]. 19. Prieto A, Garcia-Suarez J, Reyes E, Lapena P, Hernandez M, Alvarez-Mon M. Diminished DNA synthesis in T cells from B chronic lymphocytic leukemia after phytohemagglutinin, anti-CD3, and phorbol myristate acetate mitogenic signals. Exp Hematol. 1993;21:1563-1569[Medline] [Order article via Infotrieve]. 20. Cerutti A, Kim EC, Shah S, et al. Dysregulation of CD30+ T cells by leukemia impairs isotype switching in normal B cells. Nature Immunol. 2001;2:150-156[CrossRef][Medline] [Order article via Infotrieve]. 21. Trojan A, Schultze JL, Witzens M, et al. Immunoglobulin framework-derived peptides function as cytotoxic T-cell epitopes commonly expressed in B-cell malignancies. Nat Med. 2000;6:667-672[CrossRef][Medline] [Order article via Infotrieve].
22.
Harig S, Witzens M, Krackhardt AM, et al.
Induction of cytotoxic T-cell responses against immunoglobulin V region-derived peptides modified at human leukocyte antigen-A2 binding residues.
Blood.
2001;98:2999-3005
23.
Buhmann R, Nolte A, Westhaus D, Emmerich B, Hallek M.
CD40-activated B-cell chronic lymphocytic leukemia cells for tumor immunotherapy: stimulation of allogeneic versus autologous T cells generates different types of effector cells.
Blood.
1999;93:1992-2002
24.
Ranheim EA, Kipps TJ.
Activated T cells induce expression of B7/BB1 on normal or leukemic B cells through a CD40-dependent signal.
J Exp Med.
1993;177:925-935 25. Yellin MJ, Sinning J, Covey LR, et al. T lymphocyte T cell-B cell-activating molecule/CD40-L molecules induce normal B cells or chronic lymphocytic leukemia B cells to express CD80 (B7/BB-1) and enhance their costimulatory activity. J Immunol. 1994;153:666-674[Abstract]. 26. Van den Hove LE, Van Gool SW, Vandenberghe P, et al. CD40 triggering of chronic lymphocytic leukemia B cells results in efficient alloantigen presentation and cytotoxic T lymphocyte induction by up-regulation of CD80 and CD86 costimulatory molecules. Leukemia. 1997;11:572-580[CrossRef][Medline] [Order article via Infotrieve].
27.
Dorfman DM, Schultze JL, Shahsafaei A, et al.
In vivo expression of B7-1 and B7-2 by follicular lymphoma cells can prevent induction of T-cell anergy but is insufficient to induce significant T-cell proliferation.
Blood.
1997;90:4297-4306
28.
Schultze JL, Seamon MJ, Michalak S, Gribben JG, Nadler LM.
Autologous tumor infiltrating T cells cytotoxic for follicular lymphoma cells can be expanded in vitro.
Blood.
1997;89:3806-3816 29. Schultze JL, Michalak S, Seamon MJ, et al. CD40-activated human B cells: an alternative source of highly efficient antigen presenting cells to generate autologous antigen-specific T cells for adoptive immunotherapy. J Clin Invest. 1997;100:2757-2765[Medline] [Order article via Infotrieve]. 30. Kato K, Cantwell MJ, Sharma S, Kipps TJ. Gene transfer of CD40-ligand induces autologous immune recognition of chronic lymphocytic leukemia B cells. J Clin Invest. 1998;101:1133-1141[Medline] [Order article via Infotrieve]. 31. Takahashi S, Rousseau RF, Yotnda P, et al. Autologous antileukemic immune response induced by chronic lymphocytic leukemia B cells expressing the CD40 ligand and interleukin 2 transgenes. Hum Gene Ther. 2001;12:659-670[CrossRef][Medline] [Order article via Infotrieve]. 32. Banchereau J, Steinman RM. Dendritic cells and the control of immunity. Nature. 1998;392:245-252[CrossRef][Medline] [Order article via Infotrieve]. 33. Hart DN, Hill GR. Dendritic cell immunotherapy for cancer: application to low-grade lymphoma and multiple myeloma. Immunol Cell Biol. 1999;77:451-459[CrossRef][Medline] [Order article via Infotrieve]. 34. Smith JL, Barker CR. T and B cell separation by sheep-red-cell rosetting. Lancet. 1973;1:558-559[Medline] [Order article via Infotrieve]. 35. Pilling D, Kitas GD, Salmon M, Bacon PA. The kinetics of interaction between lymphocytes and magnetic polymer particles. J Immunol Methods. 1989;122:235-241[CrossRef][Medline] [Order article via Infotrieve].
36.
Romani N, Gruner S, Brang D, et al.
Proliferating dendritic cell progenitors in human blood.
J Exp Med.
1994;180:83-93 37. Brunner KT, Mauel J, Cerottini JC, Chapuis B. Quantitative assay of the lytic action of immune lymphoid cells on 51-Cr-labeled allogeneic target cells in vitro; inhibition by isoantibody and by drugs. Immunology. 1968;14:181-196[Medline] [Order article via Infotrieve]. 38. Disis ML, Gralow JR, Bernhard H, Hand SL, Rubin WD, Cheever MA. Peptide-based, but not whole protein, vaccines elicit immunity to HER-2/neu, oncogenic self-protein. J Immunol. 1996;156:3151-3158[Abstract].
39.
Sotomayor EM, Borrello I, Rattis FM, et al.
Cross-presentation of tumor antigens by bone marrow-derived antigen-presenting cells is the dominant mechanism in the induction of T-cell tolerance during B-cell lymphoma progression.
Blood.
2001;98:1070-1077
40.
Bleul CC, Wu L, Hoxie JA, Springer TA, Mackay CR.
The HIV coreceptors CXCR4 and CCR5 are differentially expressed and regulated on human T lymphocytes.
Proc Natl Acad Sci U S A.
1997;94:1925-1930
41.
Wang J, Guan E, Roderiquez G, Norcross MA.
Synergistic induction of apoptosis in primary CD4(+) T cells by macrophage-tropic HIV-1 and TGF-beta1.
J Immunol.
2001;167:3360-3366
42.
Peters W, Dupuis M, Charo IF.
A mechanism for the impaired IFN-gamma production in C-C chemokine receptor 2 (CCR2) knockout mice: role of CCR2 in linking the innate and adaptive immune responses.
J Immunol.
2000;165:7072-7077
© 2002 by The American Society of Hematology.
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
|
 |
|
  G. Gorgun, A. G. Ramsay, T. A. W. Holderried, D. Zahrieh, R. Le Dieu, F. Liu, J. Quackenbush, C. M. Croce, and J. G. Gribben E{micro}-TCL1 mice represent a model for immunotherapeutic reversal of chronic lymphocytic leukemia-induced T-cell dysfunction PNAS, April 14, 2009; 106(15): 6250 - 6255. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Galletti, C. Canones, P. Morande, M. Borge, P. Oppezzo, J. Geffner, R. Bezares, R. Gamberale, and M. Giordano Chronic Lymphocytic Leukemia Cells Bind and Present the Erythrocyte Protein Band 3: Possible Role as Initiators of Autoimmune Hemolytic Anemia J. Immunol., September 1, 2008; 181(5): 3674 - 3683. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. G. Schuster, D. H. Busch, E. Eppinger, E. Kremmer, S. Milosevic, C. Hennard, C. Kuttler, J. W. Ellwart, B. Frankenberger, E. Nossner, et al. Allorestricted T cells with specificity for the FMNL1-derived peptide PP2 have potent antitumor activity against hematologic and other malignancies Blood, October 15, 2007; 110(8): 2931 - 2939. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. P. Kater, M. H. J. van Oers, and T. J. Kipps Cellular immune therapy for chronic lymphocytic leukemia Blood, October 15, 2007; 110(8): 2811 - 2818. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Gary-Gouy, A. Sainz-Perez, J.-B. Marteau, A. Marfaing-Koka, J. Delic, H. Merle-Beral, P. Galanaud, and A. Dalloul Natural Phosphorylation of CD5 in Chronic Lymphocytic Leukemia B Cells and Analysis of CD5-Regulated Genes in a B Cell Line Suggest a Role for CD5 in Malignant Phenotype J. Immunol., October 1, 2007; 179(7): 4335 - 4344. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Palena, K. A. Foon, D. Panicali, A. G. Yafal, J. Chinsangaram, J. W. Hodge, J. Schlom, and K. Y. Tsang Potential approach to immunotherapy of chronic lymphocytic leukemia (CLL): enhanced immunogenicity of CLL cells via infection with vectors encoding for multiple costimulatory molecules Blood, November 15, 2005; 106(10): 3515 - 3523. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Hoogendoorn, J. Olde Wolbers, W. M. Smit, M. R. Schaafsma, I. Jedema, R. M.Y. Barge, R. Willemze, and J.H. F. Falkenburg Primary Allogeneic T-Cell Responses against Mantle Cell Lymphoma Antigen-Presenting Cells for Adoptive Immunotherapy after Stem Cell Transplantation Clin. Cancer Res., July 15, 2005; 11(14): 5310 - 5318. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. S. Gricks, D. Zahrieh, A. J. Zauls, G. Gorgun, D. Drandi, K. Mauerer, D. Neuberg, and J. G. Gribben Differential regulation of gene expression following CD40 activation of leukemic compared to healthy B cells Blood, December 15, 2004; 104(13): 4002 - 4009. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. von Bergwelt-Baildon, B. Maecker, J. Schultze, and J. G. Gribben CD40 activation: potential for specific immunotherapy in B-CLL Ann. Onc., June 1, 2004; 15(6): 853 - 857. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. R. Muller, G. Tsakou, F. Grunebach, S. M. Schmidt, and P. Brossart Induction of chronic lymphocytic leukemia (CLL)-specific CD4- and CD8-mediated T-cell responses using RNA-transfected dendritic cells Blood, March 1, 2004; 103(5): 1763 - 1769. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Grube, K. Rezvani, A. Wiestner, H. Fujiwara, G. Sconocchia, J. J. Melenhorst, N. Hensel, G. E. Marti, L. W. Kwak, W. Wilson, et al. Autoreactive, Cytotoxic T Lymphocytes Specific for Peptides Derived from Normal B-Cell Differentiation Antigens in Healthy Individuals and Patients with B-Cell Malignancies Clin. Cancer Res., February 1, 2004; 10(3): 1047 - 1056. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. J. M. Mackus, F. N. J. Frakking, A. Grummels, L. E. Gamadia, G. J. de Bree, D. Hamann, R. A. W. van Lier, and M. H. J. van Oers Expansion of CMV-specific CD8+CD45RA+CD27- T cells in B-cell chronic lymphocytic leukemia Blood, August 1, 2003; 102(3): 1057 - 1063. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
 |
 |
|||||||
 |
 |
 |
 |
 |
 |
 |
 |
||
| Copyright © 2002 by American Society of Hematology Online ISSN: 1528-0020 | |||||||||
Top
Abstract
Introduction
-chain of the TCR, and CD28.
