-
Djordje Atanackovic,
Ulrike Nowottne,
Eva Freier,
Cora Stefanie Weber,
Sabrina Meyer, Katrin Bartels,
York Hildebrandt,
Yanran Cao,
Nicolaus Kröger,
Monika Christine Brunner-Weinzierl,
Carsten Bokemeyer,
Hans-Christian Deter
[show abstract]
[hide abstract]
ABSTRACT: Abstract Acute psychological stress has primarily been investigated regarding its effects on conventional lymphocytes such as natural killer (NK) cells and CD4(+) and CD8(+) T cells. However, it might be important, to focus on more "specialized" lymphocyte subsets, playing a role, for instance, in allergic conditions and autoimmunity, to identify links between stress, the immune system, and somatic diseases. Using flow cytometry we determined frequencies of circulating T helper (Th)1- (CD226(+)) and Th2-type (CRTH2(+)) T cells, γδ T cells, conventional CD56(+) natural killer T (NKT) cells, and invariant NKT cells (iNKT) in healthy young males (N=31; median age 26 years) undergoing a laboratory computer-based stressor lasting 12 minutes. We found that acute psychological stress induced a prolonged increase in CD4(+) and CD8(+) T cells expressing a Th2 phenotype. We also detected an acute increase in CD4- and CD8-double negative γδ T cells. Finally, we found that the well-known increase in NK cells under stressful conditions was paralleled by a significant increase in numbers of conventional CD56(+) NKT cells. In contrast, numbers of iNKT was not altered by stress. This study adds further evidence to a psychoneuroimmunological model proposing that under stressful conditions certain lymphocyte subsets, including iNKT and less mature T cells, are retained in lymphoid tissues while antigen-experienced effector-type T cells with a Th2 phenotype, γδ T cells, and conventional CD56(+) NKT cells are mobilized into the peripheral blood. We suggest that in case of frequent stress exposure this might result in the promotion of, for example, allergic conditions.
Stress (Amsterdam, Netherlands) 02/2013; · 3.21 Impact Factor
-
Djordje Atanackovic,
York Hildebrandt,
Julia Templin,
Yanran Cao,
Christiane Keller,
Jens Panse,
Sabrina Meyer,
Henrike Reinhard, Katrin Bartels,
Nesrine Lajmi,
Orhan Sezer,
Axel R Zander,
Andreas H Marx,
Ria Uhlig,
Jozef Zustin,
Carsten Bokemeyer,
Nicolaus Kröger
[show abstract]
[hide abstract]
ABSTRACT: Multiple myeloma is a malignancy characterized by the expansion of a plasma cell clone that localizes to the human bone marrow. Myeloma cells and bone marrow stromal cells produce soluble factors that promote the survival and progression of multiple myeloma. Interleukin 16 (IL-16) is involved in regulating the migration and proliferation of normal leukocytes. However, the role of IL-16 in human cancers, including multiple myeloma, is unclear.
We investigated IL-16 expression in cell lines (n = 10) and in the bone marrow of myeloma patients (n = 62) and healthy bone marrow donors (n = 12) by quantitative reverse transcription-polymerase chain reaction, immunoblot analysis, enzyme-linked immunosorbent assay, flow cytometry, and immunohistochemistry. Transfection of two human multiple myeloma cell lines with small interfering RNAs was used to examine the effect of IL-16 gene silencing on apoptosis by flow cytometry, on proliferation by bromodeoxyuridine incorporation, and on colony formation. Protein neutralization assays were performed by treating multiple myeloma cells with a monoclonal antibody against the carboxyl-terminal fragment of IL-16. All statistical tests were two-sided.
IL-16 was strongly overexpressed in the bone marrow of myeloma patients compared with healthy donors. Myeloma cell lines as well as primary tumor cells from myeloma patients constitutively expressed IL-16 and its receptors CD4 and/or CD9 and spontaneously secreted soluble IL-16. Silencing of IL-16 reduced the proliferative activity of myeloma cells by approximately 80% compared with untreated cells (mean relative proliferative activity IL-16 siRNA vs untransfected cells, EJM cells: 20.1%, 95% confidence interval [CI] = 14.3% to 26.0%, P = .03; KMS-12-BM cells: 22.8%, 95% CI = 5.5% to 40.0%, P = .04), and addition of a recombinant carboxyl-terminal IL-16 peptide reversed that effect. A monoclonal antibody directed against IL-16 or its receptors had a comparably strong growth-inhibiting effect on the tumor cells.
IL-16 is an important growth-promoting factor in multiple myeloma and a candidate for novel diagnostic, prognostic, and therapeutic applications for this incurable human malignancy.
CancerSpectrum Knowledge Environment 06/2012; 104(13):1005-20. · 14.07 Impact Factor
-
Sebastian Kobold,
Tim Luetkens,
Britta Marlen Bartels,
Yanran Cao,
York Hildebrandt,
Orhan Sezer,
Henrike Reinhard,
Julia Templin, Katrin Bartels,
Nesrine Lajmi,
Friedrich Haag,
Carsten Bokemeyer,
Nicolaus Kröger,
Djordje Atanackovic
[show abstract]
[hide abstract]
ABSTRACT: Multiple myeloma (MM) and its therapies may induce a severely compromised humoral immunity. We have performed a longitudinal analysis of IgG-antibody responses against influenza virus (FLU) and tetanus toxoid (TT) as surrogate markers for the B cell-mediated immunity in MM patients.
1094 serum samples of 190 MM patients and samples from 100 healthy donors were analyzed by ELISA for FLU- and TT-specific antibodies.
MM patients evidenced lower levels of FLU- and TT-specific antibodies than healthy controls (P < 0.001). Immunoreactivity decreased with progressing disease and worsening clinical status. Levels of FLU- and TT-specific antibodies increased shortly (0-6 months) after alloSCT (P < 0.001), a time-period during which intravenous immunoglobulin (IVIG) is routinely applied. Thereafter, antibody concentrations declined and remained suppressed for 3 years in the case of FLU-specific and for more than 5 years in the case of TT-specific antibodies.
We found that MM is associated with a profound disease- and therapy-related immunosuppression, which is compensated for a few months after alloSCT, most likely by application of IVIG. This and the differences regarding the recovery of anti-FLU and anti-TT antibody titers during the following years need to be taken into account for optimizing IVIG application and immunization after alloSCT.
Clinical and Developmental Immunology 01/2012; 2012:134081. · 1.84 Impact Factor
-
Djordje Atanackovic,
Tim Luetkens,
Benjamin Kloth,
Gregor Fuchs,
Yanran Cao,
York Hildebrandt,
Sabrina Meyer, Katrin Bartels,
Henrike Reinhard,
Nesrine Lajmi,
Susanna Hegewisch-Becker,
Georgia Schilling,
Uwe Platzbecker,
Guido Kobbe,
Thomas Schroeder,
Carsten Bokemeyer,
Nicolaus Kröger
[show abstract]
[hide abstract]
ABSTRACT: Cancer-testis antigens (CTA) represent attractive targets for tumor immunotherapy. However, a broad picture of CTA expression in acute myeloid leukemia (AML) is missing. CTA expression was analyzed in normal bone marrow (BM) as well as in AML cell lines before and after treatment with demethylating agents and/or histone acetylase inhibitors. Presence of selected CTA with a strictly tumor-restricted expression was then determined in samples of patients with AML before and after demethylating therapy. Screening AML cell lines for the expression of 20 CTA, we identified six genes (MAGE-A3, PRAME, ROPN1, SCP-1, SLLP1, and SPO11) with an AML-restricted expression. Analyzing the expression of these CTA in blast-containing samples from AML patients (N = 64), we found all samples to be negative for MAGE-A3 and SPO11 while a minority of patients expressed ROPN1 (1.6%), SCP-1 (3.1%), or SLLP1 (9.4%). The only CTA expressed in substantial proportion of patients (53.1%) was PRAME. Following demethylating treatment with 5'-aza-2'-deoxycytidine, we observed an increased or de novo expression of CTA, in particular of SSX-2, in AML cell lines. In AML patients, we detected increased expression of PRAME and induction of SSX-2 after demethylating therapy with 5-azacytidine. With the exception of PRAME, CTA are mostly absent from AML blasts. However, demethylating treatment induces strong expression of CTA, particularly of SSX-2, in vitro and in vivo. Therefore, we propose that CTA-specific immunotherapy for AML should preferentially target PRAME and/or should be combined with the application of demethylating agents opening the perspective for alternative targets like CTA SSX-2.
American Journal of Hematology 07/2011; 86(11):918-22. · 4.67 Impact Factor
-
Djordje Atanackovic,
Jens Panse,
York Hildebrandt,
Adam Jadczak,
Sebastian Kobold,
Yanran Cao,
Julia Templin,
Sabrina Meyer,
Henrike Reinhard, Katrin Bartels,
Nesrine Lajmi,
Axel R Zander,
Andreas H Marx,
Carsten Bokemeyer,
Nicolaus Kröger
[show abstract]
[hide abstract]
ABSTRACT: To date, multiple myeloma remains an incurable malignancy due to the persistence of minimal residual disease in the bone marrow. In this setting, monoclonal antibodies against myeloma-specific cell surface antigens represent a promising therapeutic approach, which is however hampered by a lack of appropriate target structures expressed across all pathogenic myeloma cell populations. We, therefore, investigated functionally relevant immunoreceptors specifically associated with myeloma cells as well as their clonogenic precursors.
Potential target proteins were identified using antibody arrays against phosphorylated immunoreceptors with lysates from myeloma cell lines. CD229 expression was confirmed in primary myeloma cells by reverse transcriptase polymerase chain reaction, western blot, fluorescence-activated cell sorting, and immunohistochemistry. Apoptosis, clonogenic growth, and sensitivity to chemotherapy were determined following short-interfering RNA-mediated downregulation of CD229. Antibody-dependent cellular and complement-dependent cytotoxicity were analyzed using a monoclonal antibody against CD229 to demonstrate the antigen's immunotherapeutic potential.
Our screening assay identified CD229 as the most strongly over-expressed/phosphorylated immunoreceptor in myeloma cell lines. Over-expression was further demonstrated in the CD138-negative population, which has been suggested to represent myeloma precursors, as well as on primary tumor cells from myeloma patients. Accordingly, CD229 staining of patients' bone marrow samples enabled the identification of myeloma cells by flow cytometry and immunohistochemistry. Down-regulation of CD229 led to a decreased number of viable myeloma cells and clonal myeloma colonies, and enhanced the anti-tumor activity of conventional chemotherapeutics. Targeting CD229 with a monoclonal antibody resulted in complement- and cell-mediated lysis of myeloma cells.
Our results demonstrate that the immunoreceptor CD229 is specifically over-expressed on myeloma cells including their clonogenic precursors and contributes to their malignant phenotype. Monoclonal antibodies against this protein may represent a promising diagnostic and immunotherapeutic instrument in this disease.
Haematologica 05/2011; 96(10):1512-20. · 6.42 Impact Factor
-
Sebastian Kobold,
Sinje Tams,
Tim Luetkens,
Yanran Cao,
Orhan Sezer,
Britta Marlen Bartels,
Henrike Reinhard,
Julia Templin, Katrin Bartels,
York Hildebrandt,
Nesrine Lajmi,
Andreas Marx,
Friedrich Haag,
Carsten Bokemeyer,
Nicolaus Kröger,
Djordje Atanackovic
[show abstract]
[hide abstract]
ABSTRACT: The occurrence of SOX2-specific autoantibodies seems to be associated with an improved prognosis in patients with monoclonal gammopathy of undetermined significance (MGUS). However, it is unclear if SOX2-specific antibodies also develop in established multiple myeloma (MM). Screening 1094 peripheral blood (PB) sera from 196 MM patients and 100 PB sera from healthy donors, we detected SOX2-specific autoantibodies in 7.7% and 2.0% of patients and donors, respectively. We identified SOX2(211-230) as an immunodominant antibody-epitope within the full protein sequence. SOX2 antigen was expressed in most healthy tissues and its expression did not correlate with the number of BM-resident plasma cells. Accordingly, anti-SOX2 immunity was not related to SOX2 expression levels or tumor burden in the patients' BM. The only clinical factor predicting the development of anti-SOX2 immunity was application of allogeneic stem cell transplantation (alloSCT). Anti-SOX2 antibodies occurred more frequently in patients who had received alloSCT (n = 74). Moreover, most SOX2-seropositive patients had only developed antibodies after alloSCT. This finding indicates that alloSCT is able to break tolerance towards this commonly expressed antigen. The questions whether SOX2-specific autoantibodies merely represent an epiphenomenon, are related to graft-versus-host effects or participate in the immune control of myeloma needs to be answered in prospective studies.
Clinical and Developmental Immunology 01/2011; 2011:302145. · 1.84 Impact Factor
-
Tim Luetkens,
Phillipe Schafhausen,
Frederike Uhlich,
Tim Stasche,
Ruken Akbulak,
Britta M Bartels,
York Hildebrandt,
Arthur Gontarewicz,
Sebastian Kobold,
Sabrina Meyer,
Maja Gordic, Katrin Bartels,
Nesrine Lajmi,
Yanran Cao,
Nicolaus Kröger,
Carsten Bokemeyer,
Tim H Brümmendorf,
Djordje Atanackovic
[show abstract]
[hide abstract]
ABSTRACT: Cancer-testis (CT) antigens represent attractive targets for tumor immunotherapy based on their tumor-restricted expression and immunogenicity. However, a broad picture of the expression of CT antigens and associated humoral immune responses in chronic myeloid leukemia (CML) is still missing.
We screened CML cell lines and bone marrow (BM) samples from healthy donors by RT-PCR for the expression of 31 CT antigens before and after treatment with epigenetic agents. Expression of tumor-restricted antigens was further examined in 60 CML patients and humoral immune responses against 15 CT antigens were screened by ELISA.
In untreated cell lines we detected the expression of 17 CT antigens that were absent from normal BM. Expression of most antigens increased following demethylating treatment with 5'-Aza-2'-Deoxycytidine. In these samples, only PRAME was repeatedly detected and expression correlated with several clinicopathological parameters and decreased overall survival. We further show that a lower frequency of PRAME-positive samples during imatinib treatment was not caused by gene-specific downregulation. Analyzing the patients' antibody responses we found that the vast majority of patients lacked spontaneous immunity against CT antigens including PRAME.
CT antigen expression can be increased by the application of epigenetic agents and the expression of PRAME correlates with clinicopathological parameters and overall survival in patients with CML, but does not lead to humoral immune responses. PRAME-specific immunotherapy might represent a promising approach for the eradication of residual therapy-resistant leukemic cells due to its frequent expression and stability under imatinib treatment.
Leukemia research 12/2010; 34(12):1647-55. · 2.36 Impact Factor
-
Yanran Cao,
Tim Luetkens,
Sebastian Kobold,
York Hildebrandt,
Maja Gordic,
Nesrine Lajmi,
Sabrina Meyer, Katrin Bartels,
Axel R Zander,
Carsten Bokemeyer,
Nicolaus Kröger,
Djordje Atanackovic
[show abstract]
[hide abstract]
ABSTRACT: The interaction of multiple myeloma (MM) with its bone marrow (BM) microenvironment is important for the homing pattern, survival, and proliferation of malignant plasma cells. We aimed at answering the question which cytokines, chemokines, and growth factors are typically found in the BM of untreated MM patients as well as in MM patients after allogeneic stem cell transplantation (alloSCT).
We determined the concentrations of 34 cytokines/chemokines in the supernatants of 10 myeloma cell lines, as well as in the plasma derived from BM and peripheral blood samples of 10 newly diagnosed MM patients, 20 MM patients who had received allogeneic stem cell transplantation (alloSCT), and 20 healthy donors.
Besides cytokines/chemokines known to be secreted by myeloma cell lines, such as interleukin-1 receptor antagonist (IL-1RA), IL-8, monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein (MIP)-1α, MIP-1β, and MIP-3α, we also detected significant levels of epidermal growth factor (EGF), hepatocyte growth factor (HGF), IL2R, IL-12p40/p70, IL-22, interferon-γ (IFN-γ)-inducible protein 10 (IP-10), monokine induced by IFN-γ (MIG), and regulated on activation normally T-cell expressed and secreted (RANTES) in culture supernatants. The BM environment in MM patients evidenced elevated concentrations of HGF, IL-2R, IL-16, EGF, IL-1RA, IP-10, MCP-1, and monokine induced by IFN-γ. Additionally, in the BM of MM patients post alloSCT, we found selectively elevated concentration of IL-4, IL-6, IL-8, IL-12p40/p70, and eotaxin. Eotaxin levels were particularly high in patients with chronic graft-vs-host disease.
Our study demonstrates characteristic cytokine/chemokine patterns in the BM environment of MM patients before and after alloSCT. Certain factors, such as MIP-1α, MCP-1, HGF, IL-16, IP-10, and eotaxin, might not only be developed into diagnostic instruments and/or predictive biomarkers, but are also potential targets for future myeloma- or graft-vs-host disease-specific therapies.
Experimental hematology 10/2010; 38(10):860-7. · 3.11 Impact Factor
-
Yanran Cao,
Maja Gordic,
Sebastian Kobold,
Nesrine Lajmi,
Sabrina Meyer, Katrin Bartels,
York Hildebrandt,
Tim Luetkens,
Anna Sophie Ihloff,
Nicolaus Kröger,
Carsten Bokemeyer,
Djordje Atanackovic
[show abstract]
[hide abstract]
ABSTRACT: In the framework of our current study we set out to develop an optimized assay for the quantification of antigen-specific B cells in different compartments of the human body.
Mononuclear cells (MNC) derived from the peripheral blood, bone marrow (BM), or human tonsils were incubated with different combinations of stimuli. The stimulated cells and culture supernatants were then applied to IgG-ELISPOT and ELISA read-out assays and tetanus toxoid (TT)-specific B cell responses were quantified.
We found that a combination of CD40L, CpG, and IL21 was optimal for the induction of TT-specific IgG-producing cells from memory B cell (mBc) precursors. This cocktail of stimuli led to a proliferation-dependent induction of CD19(intermediate)CD38(high)CD138(high)IgD(negative) terminally differentiated plasma cells. Applying our optimized methodology we were also able to quantify mBc specific for cytomegalovirus and influenza virus A. Most importantly, the same method proved useful for the comparison of mBc frequencies between different compartments of the body and, accordingly, we were able to demonstrate that TT-specific mBc preferably reside within tonsillar tissue.
Here, we optimized an assay for the quantification of antigen-specific B cells in different human tissues demonstrating, for example, that TT-specific mBc preferably reside in human tonsils but not in the BM or the peripheral blood. We suggest that our approach can be used for the enumeration of mBc specific for a wide variety of Ag (microbial, tumor-related, auto-antigens), which will lead to significant improvements regarding our knowledge about the biology of humoral immunity.
Journal of immunological methods 03/2010; 358(1-2):56-65. · 2.35 Impact Factor
-
Djordje Atanackovic,
York Hildebrandt,
Adam Jadczak,
Yanran Cao,
Tim Luetkens,
Sabrina Meyer,
Sebastian Kobold, Katrin Bartels,
Caroline Pabst,
Nesrine Lajmi,
Maja Gordic,
Tanja Stahl,
Axel R Zander,
Carsten Bokemeyer,
Nicolaus Kröger
[show abstract]
[hide abstract]
ABSTRACT: Multiple myeloma is a life-threatening disease and despite the introduction of stem cell transplantation and novel agents such as thalidomide, lenalidomide, and bortezomib most patients will relapse and develop chemoresistant disease. Therefore, alternative therapeutic modes for myeloma are needed and cancer-testis antigens such as MAGE-C1/CT7 and MAGE-A3 have been suggested to represent a class of tumor-specific proteins particularly suited for targeted immunotherapies. Surprisingly, the biological role of cancer-testis genes in myeloma remains poorly understood.
We performed the first investigation of the function of two cancer-testis antigens most commonly expressed in myeloma, MAGE-C1/CT7 and MAGE-A3, using an RNA interference-based gene silencing model in myeloma cell lines. Functional assays were used to determine changes in proliferation, cell adhesion, chemosensitivity, colony formation, and apoptosis resulting from gene-specific silencing.
We show that the investigated genes are not involved in regulating cell proliferation or adhesion; however, they play an important role in promoting the survival of myeloma cells. Accordingly, knock-down of MAGE-C1/CT7 and MAGE-A3 led to the induction of apoptosis in the malignant plasma cells and, importantly, both genes were also essential for the survival of clonogenic myeloma precursors. Finally, silencing of cancer-testis genes further improved the response of myeloma cells to conventional therapies.
Cancer-testis antigens such as MAGE-C1/CT7 and MAGE-A3 play an important role in promoting the survival of myeloma cells and clonogenic precursors by reducing the rate of spontaneous and chemotherapy-induced apoptosis and might, therefore, represent attractive targets for novel myeloma-specific therapies.
Haematologica 12/2009; 95(5):785-93. · 6.42 Impact Factor
-
Eva Freier,
Cora Stefanie Weber,
Ulrike Nowottne,
Christiane Horn, Katrin Bartels,
Sabrina Meyer,
York Hildebrandt,
Tim Luetkens,
Yanran Cao,
Caroline Pabst,
Julia Muzzulini,
Benjamin Schnee,
Monika Christine Brunner-Weinzierl,
Maurizio Marangolo,
Carsten Bokemeyer,
Hans-Christian Deter,
Djordje Atanackovic
[show abstract]
[hide abstract]
ABSTRACT: We have previously shown that acute psychological stress alerts the adaptive immune response causing an increase in antigen-experienced effector T cells in the peripheral blood. T regulatory cells (Tregs) play a central role in maintaining self-tolerance and controlling autoimmune responses. Here, we analyzed for the first time the behaviour of Tregs in the context of a stress-induced activation of the adaptive immune response. 31 healthy young males underwent a brief laboratory stressor and, in a crossover design, served as their own unstressed controls. We quantified effects of acute stress on CD4(+)FOXP3(+) T regulatory cells and other T cell subpopulations using flow cytometry. In addition, the expression of Treg-related effector molecules and stress hormone receptors were analyzed in the subjects' peripheral T cells. We confirmed our previous observation of a stress-induced decrease in CD45RA(+)CCR7(+) "naïve" and CD45RA(-)CCR7+ "central memory" T cells while CD45RA(-)-CCR7(-) "memory effector" and CD45RA(+)CCR7(-) "terminally differentiated" effector T cells remained stable or increased. Importantly, we found acute psychological stress to cause a concomitant decrease in CD4(+)FOXP3(+) Tregs and in CD4(+) T cells expressing Treg-related effector molecules cytotoxic T-lymphocyte antigen-4 (CTLA-4) and latency associated peptide (LAP). Finally, we observed beta(1)-adrenergic and glucorticoid alpha receptors to be overexpressed in Tregs, suggesting that these molecules might mediate stress-related effects on Tregs. In conclusion, inhibiting components of the adaptive immune response, like Tregs, are down-regulated during a stress-induced activation of the adaptive immune response. In situations of chronic stress, this scenario might result in an exacerbation of inflammatory conditions such as autoimmune diseases.
Psychoneuroendocrinology 12/2009; 35(5):663-73. · 5.81 Impact Factor
-
Sacha Gnjatic,
Yanran Cao,
Uta Reichelt,
Emre F Yekebas,
Christina Nölker,
Andreas H Marx,
Andreas Erbersdobler,
Hiroyoshi Nishikawa,
York Hildebrandt, Katrin Bartels, [......],
Vincenzo Cerundolo,
Tim Luetkens,
Gerd Ritter,
Kay Friedrichs,
Rudolf Leuwer,
Susanna Hegewisch-Becker,
Jakob R Izbicki,
Carsten Bokemeyer,
Lloyd J Old,
Djordje Atanackovic
[show abstract]
[hide abstract]
ABSTRACT: NY-CO-58/KIF2C has been identified as a tumor antigen by screening antibody responses in patients with colorectal cancer. However, expression had not consequently been examined, and nothing was known about its ability to induce spontaneous T cell responses, which have been suggested to play a role in the development of colorectal cancer. We analyzed 5 colorectal cancer cell lines, and tumor samples and adjacent healthy tissues from 176 patients with epithelial cancers for the expression of NY-CO-58/KIF2C by RT-PCR and Western Blot. T cell responses of 43 colorectal cancer patients and 35 healthy donors were evaluated by ELISpot following stimulation with 30mer peptides or full-length protein. All cell lines and tumor samples from colorectal cancer patients expressed NY-CO-58/KIF2C on the protein and RNA level, and expression levels correlated strongly with Ki-67 expression (r = 0.69; p = 0.0003). Investigating NY-CO-58/KIF2C-specific T cell responses, CD8(+) T cells directed against 1 or more peptides were found in less than 10% of patients, whereas specific CD4(+) T cells were detected in close to 50% of patients. These T cells were of high avidity, recognized the naturally processed antigen and secreted IFN-gamma and TNF-alpha. Depletion of CD4(+)CD25(+) T cells before stimulation significantly increased the intensity of the preexisting response. NY-CO-58/KIF2C is significantly overexpressed in colorectal and other epithelial cancers and expression levels correlate with the proliferative activity of the tumor. Importantly, NY-CO-58/KIF2C was able to induce spontaneous CD4(+) T cell responses of the Th1-type, which were tightly controlled by peripheral T regulatory cells.
International Journal of Cancer 11/2009; 127(2):381-93. · 5.44 Impact Factor
-
Djordje Atanackovic,
Tim Luetkens,
York Hildebrandt,
Julia Arfsten, Katrin Bartels,
Christiane Horn,
Tanja Stahl,
Yanran Cao,
Axel R Zander,
Carsten Bokemeyer,
Nicolaus Kröger
[show abstract]
[hide abstract]
ABSTRACT: Reliable data on the persistence of tumor expression of cancer-testis (CT) antigens over time and consequent analyses of the effect of CT antigen expression on the clinical course of malignancies are crucial for their evaluation as diagnostic markers and immunotherapeutic targets.
Applying conventional reverse transcription-PCR, real-time PCR, and Western blot, we did the first longitudinal study of CT antigen expression in multiple myeloma analyzing 330 bone marrow samples from 129 patients for the expression of four CT antigens (MAGE-C1/CT7, MAGE-C2/CT10, MAGE-A3, and SSX-2).
CT antigens were frequently and surprisingly persistently expressed, indicating that down-regulation of these immunogenic targets does not represent a common tumor escape mechanism in myeloma. We observed strong correlations of CT antigen expression levels with the clinical course of myeloma patients as indicated by the number of bone marrow-residing plasma cells and peripheral paraprotein levels, suggesting a role for CT antigens as independent tumor markers. Investigating the prognostic value of CT antigen expression in myeloma patients after allogeneic stem cell transplantation, we found that expression of genes, such as MAGE-C1, represents an important indicator of early relapse and dramatically reduced survival.
Our findings suggest that CT antigens might promote the progression of multiple myeloma and especially MAGE-C1/CT7, which seems to play the role of a "gatekeeper" gene for other CT antigens, might characterize a more malignant phenotype. Importantly, our study also strongly supports the usefulness of CT antigens as diagnostic and prognostic markers as well as therapeutic targets in myeloma.
Clinical Cancer Research 03/2009; 15(4):1343-52. · 7.74 Impact Factor
-
Djordje Atanackovic,
Yanran Cao,
Tim Luetkens,
Jens Panse,
Christiane Faltz,
Julia Arfsten, Katrin Bartels,
Christine Wolschke,
Thomas Eiermann,
Axel R Zander,
Boris Fehse,
Carsten Bokemeyer,
Nicolaus Kroger
[show abstract]
[hide abstract]
ABSTRACT: Very little is known about the number and function of immunosuppressive CD4(+)CD25(+)FOXP3(+) T regulatory cells (Treg) in the human bone marrow and it is unclear whether bone marrow-residing Treg are capable of regenerating following allogeneic stem cell transplantation. This is particularly surprising since the bone marrow represents a major priming site for T-cell responses and Treg play important roles in the prevention of T-cell-mediated graft-versus-host disease and in promoting tumor escape from T-cell-dependent immunosurveillance.
Applying flow cytometry, real-time polymerase chain reaction, and functional assays, we performed the first study on bone marrow and peripheral blood Treg in healthy donors as well as multiple myeloma patients before and after allogeneic stem cell transplantation.
We found that, following the allogeneic transplantation, donor-derived CD4(+)CD25(+)FOXP3(+) Treg expanded faster than conventional CD4(+) T cells, leading to an accumulation of Treg in the bone marrow of transplanted patients who lack relevant thymic function. The reconstituted bone marrow-residing CD4(+)CD25(+)FOXP3(+) Treg of myeloma patients after allogeneic stem cell transplantation consisted preferably of CD45RA(-)CCR7(-) memory T-cells and contained low numbers of T-cell receptor excision cycles, indicating that Treg had indeed expanded outside the thymus. Importantly, bone marrow-residing Treg of newly diagnosed and myeloma patients after allogeneic stem cell transplantation expressed high levels of transforming growth factor beta and cytotoxic T-lymphocyte antigen 4, and showed a strong inhibitory function.
We suggest that allogeneic stem cell transplantation provides a short but significant window of opportunity for CD8(+) T cells before an exuberant regeneration of immunosuppressive Treg sets in. Later after transplantation, bone marrow-residing Treg probably contribute to suppressing graft-versus-host disease but may also undermine persistent immune control of multiple myeloma.
Haematologica 04/2008; 93(3):423-30. · 6.42 Impact Factor
-
Djordje Atanackovic,
Yanran Cao,
Ji-Won Kim,
Stephan Brandl,
Ina Thom,
Christiane Faltz,
York Hildebrandt, Katrin Bartels,
Andreas de Weerth,
Susanna Hegewisch-Becker,
Dieter Kurt Hossfeld,
Carsten Bokemeyer
[show abstract]
[hide abstract]
ABSTRACT: Malignant effusions offer a unique opportunity for the study of interactions between the human immune system and cancer. We have recently demonstrated that malignant effusions are characterized by an accumulation of T cells expressing chemokine receptors such as CCR4, which is commonly found on Th2 cells. In contrast, effector T cells expressing chemokine receptors typical for Th1 cells, such as CCR5, showed a diminished homing into malignant effusions.
We analyzed concentrations of 12 different cytokines and 9 chemokines within malignant and nonmalignant effusions and investigated cytokine expression by effusion-infiltrating leukocytes.
We observed that concentrations of the immunoregulatory cytokine TGF-beta(1) and of angiogenic factors VEGF and IL-8 were markedly increased within effusions caused by malignancies. However, we did not observe signs of a typical Th1 or Th2 milieu. Analyzing concentrations of 9 different chemokines, we found elevated concentrations of the chemokines MDC, eotaxin, I-TAC, and MCP-1 in malignant effusions. Interestingly, tumor-infiltrating leukocytes themselves seemed to contribute strongly to the creation of a distinct cytokine/chemokine pattern within cancer-related effusions. Additional analyses suggested that this cytokine/chemokine milieu might support an enrichment of immunosuppressive leukocytes.
The local cytokine and chemokine milieu within malignant effusions seems to promote angiogenesis and to block an efficient immune-mediated antitumor response. An elimination of such tumor-promoting influences will be necessary in order to transform local immunotolerance into clinically relevant immune recognition of tumors causing malignant effusions.
Tumor Biology 02/2008; 29(2):93-104. · 1.94 Impact Factor
-
Djordje Atanackovic,
Julia Arfsten,
Yanran Cao,
Sacha Gnjatic,
Frank Schnieders, Katrin Bartels,
Georgia Schilling,
Christiane Faltz,
Christine Wolschke,
Judith Dierlamm,
Gerd Ritter,
Thomas Eiermann,
Dieter Kurt Hossfeld,
Axel R Zander,
Achim A Jungbluth,
Lloyd J Old,
Carsten Bokemeyer,
Nicolaus Kröger
[show abstract]
[hide abstract]
ABSTRACT: Immunotherapies using cancer-testis (CT) antigens as targets represent a potentially useful treatment in patients with multiple myeloma (MM) who commonly show recurrent disease following chemotherapy. We analyzed the expression of 11 CT antigens in bone marrow samples from patients with MM (n=55) and healthy donors (n=32) using reverse transcriptase-polymerase chain reaction (RT-PCR). CT antigens were frequently expressed in MM with 56% (MAGEC2), 55% (MAGEA3), 35% (SSX1), 20% (SSX4, SSX5), 16% (SSX2), 15% (BAGE), 7% (NY-ESO-1), and 6% (ADAM2, LIPI) expressing the given antigen. Importantly, CT antigens were not expressed in healthy bone marrow. Analyzing patients with MM (n=66) for antibody responses against MAGEA3, SSX2, and NY-ESO-1, we found strong antibody responses against CT antigens preferentially in patients who had received allogeneic stem cell transplantation (alloSCT). Antibody responses against NY-ESO-1 correlated with NY-ESO-1-specific CD4+ and CD8+ T-cell responses against peptide NY-ESO-1(51-62) and CD4+ responses against NY-ESO-1(121-140) in 1 of these patients. These allogeneic immune responses were not detectable in pretransplantation samples and in the patients' stem cell donors, indicating that CT antigens might indeed represent natural targets for graft-versus-myeloma effects. Immune responses induced by alloSCT could be boosted by active CT antigen-specific immunotherapy, which might help to achieve long-lasting remissions in patients with MM.
Blood 03/2007; 109(3):1103-12. · 9.90 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Cancer-Testis (CT) antigens are by definition expressed in tumor but not in healthy tissue except testis and might represent ideal targets for antigen-specific immunotherapy. Here, we present the first comprehensive analysis of CT antigen expression in patients with head and neck squamous cell carcinoma (HNSCC). Tumor samples (N = 51), and adjacent healthy tissue from patients with HNSCC were analyzed for the expression of 23 genes designated CT antigens using RT-PCR. Patient sera (N = 39) were screened for IgG antibody responses against NY-ESO-1, MAGEA3, and SSX2. According to their expression pattern antigens were divided into four groups. ADAM2, LIP1, SLLP1, AKAP3, CTAGE, ZNF165, CAGE, and FTHL17 were expressed in tumor and healthy tissue at comparable frequencies. NY-TLU-57, GAGE1, SAGE1 were expressed more frequently in tumor samples than in healthy tissues. TPTE, LDHC, SPO11 were expressed neither in tumor samples nor in healthy tissue. 9 CT antigens were expressed only in the tumor tissue and may represent ideal candidates for active immunotherapy in HNSCC: MAGEA3 was expressed in 72%, SSX1 in 45%, MAGEC2 in 33%, MAGEC1 in 28%, BAGE in 17%, SSX2 in 16%, SCP1 in 12%, NY-ESO-1 in 6%, and HOM-TES-85 in 4% of cases. 86% of tumor samples expressed at least one, 69% expressed at least two, and 43% expressed at least three of these antigens. Three patients showed an antibody response against NY-ESO-1. In conclusion, we demonstrate here that HNSCC frequently express CT antigens. Furthermore, a relatively high percentage of tumors express more than one CT antigen opening the perspective for polyvalent antigen-specific immunotherapy.
Cancer biology & therapy 10/2006; 5(9):1218-25. · 2.64 Impact Factor
-
Djordje Atanackovic,
Benjamin Schnee,
Gunter Schuch,
Christiane Faltz,
Julia Schulze,
Cora S Weber,
Philippe Schafhausen, Katrin Bartels,
Carsten Bokemeyer,
Monika Christine Brunner-Weinzierl,
Hans-Christian Deter
[show abstract]
[hide abstract]
ABSTRACT: Influences of psychological stress on the acquired immune system have not consequently been investigated. We found acute psychological stress to cause an increase in CD56+ and CCR5+ effector T cells in the peripheral blood of healthy human subjects (N=22), while skin-homing CLA+ T cells decreased. At the same time, we observed a stress-induced decrease in CD45RA+/CCR7+ naive and CD45RA-/CCR7+ central memory T cells, while CD45RA-/CCR7- effector memory and CD45RA+/CCR7- terminally differentiated T cells increased. This T cell redistribution translated into an increase in T cells expressing perforin/granzyme B and in Epstein-Barr virus-specific, cytomegalovirus-specific and influenza virus-specific CD8+ T cells. Thus, acute stress seems to promote the retention of less mature T cells within lymphoid tissue or skin while effector-type T cells are mobilized into the blood in order to be able to rapidly migrate into peripheral tissues.
Journal of Neuroimmunology 08/2006; 176(1-2):141-52. · 2.96 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Tumor-infiltrating T cells have a positive influence on the clinical course of B cell non-Hodgkin's lymphoma (NHL). T cells in the peripheral blood of patients with B cell NHL, however, have so far rarely been examined.
Using flow cytometry we examined lymphocyte subpopulations and numbers of naïve/memory T cell subtypes among peripheral T cells of patients with B cell NHL (N=22), patients with metastasized solid tumors (N=27), and healthy controls (N=20). In addition, we analyzed the intracellular content of effector molecules granzyme B and perforin and expression of the T cell receptor zeta chain.
We observed increased percentages of potentially highly cytotoxic CD8+CD56+ T cells in the peripheral blood of patients with NHL. Both, patients with NHL and patients with solid tumors showed a much higher expression of the chemokine receptors CCR4 and CCR5 on their T cells than healthy controls, suggesting a polarization of their T cells following stimulation with antigen and/or cytokines in vivo. Furthermore, patients with B cell NHL and patients with solid tumors had far lower percentages of naïve CD45RA+CCR7+ T cells than healthy controls and, in the case of CD4+ T cells, patients with solid tumors. In contrast, patients with B cell NHL showed markedly increased levels of memory effector CD45RA-CCR7- CD4(+) T cells when compared to healthy controls and patients with metastasized solid tumors. Patients with NHL also showed elevated levels granzyme B within CD8(+) T cells, indicating that the increase in memory effector cells was of functional relevance.
These findings indicate a marked shift in the composition of peripheral T cells of patients with B cell NHL from naïve to memory effector-type cells.
Leukemia Research 10/2005; 29(9):1019-27. · 2.92 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Diabetes-prone BB (dpBB) rats develop autoimmune insulin-dependent diabetes mellitus (IDDM) at high frequency as a consequence of a defect in T cell development, caused by a mutation in a single gene locus on rat chromosome 4 (lyp) which has recently been identified as immune-associated nucleotide 4 (ian4). A phenotype similar to dpBB rat lymphopenia has recently been described in the mouse as the result of the targeted inactivation of the gene for the transcription factor LKLF (Lung Krüppel-like factor, KLF2) in the immune system. We cloned the LKLF gene of the rat and screened a panel of rat/hamster radiation hybrid cell lines to determine its chromosomal localization. We conclude that the LKLF gene is not defective in dpBB rats and that its expression is not compromised by the lyp mutation.
Annals of the New York Academy of Sciences 01/2004; 1010:548-51. · 3.15 Impact Factor
