Dieter Kabelitz - Profile on Academia.edu (original) (raw)
Papers by Dieter Kabelitz
Journal of Immunology, Jun 1, 2006
The oncofetal Ag immature laminin receptor (OFA-iLR) is a potential target molecule for immunothe... more The oncofetal Ag immature laminin receptor (OFA-iLR) is a potential target molecule for immunotherapeutic studies in several tumor entities, including hematological malignancies. In the present study, we characterize two HLA-A*0201-presented epitopes eliciting strong OFA-iLR peptide-specific human cytotoxic T cell (CTLs) responses in vitro. Both allogeneic HLA-A*0201-matched and autologous CTLs recognized and killed endogenously OFA-iLR-expressing tumor cell lines and primary malignant cells from patients with hemopoietic malignancies in an MHC-restricted fashion but spared nonmalignant hemopoietic cells. Spontaneous OFA-iLR peptide-specific T cell reactivity was detectable in a significant proportion of leukemia patients. Interestingly, in patients with chronic lymphocytic leukemia and multiple myeloma but not in those with acute myeloid leukemia, significant frequencies of OFA peptide-specific CTLs could be detected in an early stage of disease but disappeared in patients with progressive disease. The identification of OFA-iLR-derived peptide epitopes provides a basis for tumor immunological studies and therapeutic vaccination strategies in patients with OFA-iLR-expressing malignancies.
Frontiers in Immunology, Sep 30, 2022
gamma delta (gd) T cells, tumor immunotherapy, butyrophilins, cytokine programming, gamma delta T... more gamma delta (gd) T cells, tumor immunotherapy, butyrophilins, cytokine programming, gamma delta TCR repertoire, cost-effective manufacturing of gamma delta t cell therapies, chimeric antigen receptor-engineered gamma delta T cells, phosphoantigens The discovery of the gd T-cell receptor (TCR) and its ability to confer potent cytotoxic activity in CD3+ cells some 35 years ago sparked the initial proliferation in research that garnered widespread interest in the biology and function of gd T cells. The identification of major human gd TCR clonotypes, their tissue distributions as well as dynamic changes throughout ontogeny and in disease states have contributed to the appreciation of human gd T cell diversity. Despite this, incomplete understanding of mechanisms underlying the complexity of various gd T cell subsets in homeostasis, inflammation and malignancy restricted the focus of most early studies to blood circulating Vg9Vd2 T cells which could be robustly activated and expanded ex vivo using aminobisphosphonates such as zoledronate compared with other gd T cell subsets for which activating ligands were then largely unknown. This galvanized attempts to harness the tumoricidal potential of Vg9Vd2 T cells in clinical trials. Although these cells exhibited highly promising safety profiles in patients, early trial data revealed suboptimal anti-tumor efficacy. Despite these setbacks, recent breakthroughs in deciphering the unique antigen (Ag) binding modes of gd TCR coupled with high dimensional analyses of tissue-and disease-specific gd T cell subsets at single cell resolution led to renewed excitement in the development of gd T cell-based therapeutics. This Research Topic has compiled a series of nine articles of which five review our hitherto understanding of the multifaceted nature of gd T cells and four report original research providing new insights into the molecular and cellular regulation of a diverse repertoire of gd T cells, paving the way for next generation gd T cell-based tumor immunotherapies.
Frontiers in Immunology, Nov 12, 2021
Vitamin C (VitC), in addition to its role as a general antioxidant, has long been considered to p... more Vitamin C (VitC), in addition to its role as a general antioxidant, has long been considered to possess direct anti-cancer activity at high doses. VitC acts through oxidant and epigenetic mechanisms, which at high doses can exert direct killing of tumor cells in vitro and delay tumor growth in vivo. Recently, it has also been shown that pharmacologicdose VitC can contribute to control of tumors by modulating the immune system, and studies have been done interrogating the role of physiologic-dose VitC on novel adoptive cellular therapies (ACTs). In this review, we discuss the effects of VitC on anti-tumor immune cells, as well as the mechanisms underlying those effects. We address important unanswered questions concerning both VitC and ACTs, and outline challenges and opportunities facing the use of VitC in the clinical setting as an adjunct to immune-based anti-cancer therapies. Keywords: immune checkpoint therapy (ICT), CAR (chimeric antigen receptor) T cells, vitamin C (ascorbic acid), cancer immunotherapies, cancer biology
The Journal of Immunology
We have investigated the frequency and specificity of gamma delta+ cytotoxic lymphocyte precursor... more We have investigated the frequency and specificity of gamma delta+ cytotoxic lymphocyte precursors (CLP) under limiting dilution culture conditions. E rosette separated total T cells and CD3+CD4-CD8-TCR alpha beta- double-negative (DN) T cells were cocultured with allogeneic or autologous PBMC stimulator cells, and frequencies of alloreactive and autoreactive CLP were determined after 12 to 14 days against Con A blast target cells. Freshly isolated DN cells consisting of 82.3 +/- 8.2% gamma delta+ T cells did not exert cytolytic activity against K562 or anti-TCR gamma delta mAb-producing hybridoma cells. In striking contrast to E+ cells, the vast majority of alloantigen-stimulated clonally developing DN CLP did not show specificity for stimulator-derived target cells. Thus, frequencies of alloreactive and autoreactive CLP after alloantigenic stimulation were in the range of 1/100 to 1/4800 and 1/450 to 1/5000, respectively. After coculture with autologous stimulator cells, frequenci...
The Journal of Immunology
It is well established that IL-4 and IL-7 control the differentiation of TCR-gamma delta-expressi... more It is well established that IL-4 and IL-7 control the differentiation of TCR-gamma delta-expressing cells from CD3/TCR-negative thymic precursors. In this study, we have compared the in situ expression of IL-4 and IL-7 mRNA in human postnatal thymus with the in vitro effect of IL-4 and IL-7 on the expansion of TCR-gamma delta + cells from highly purified CD3-4-8- triple-negative thymocytes. IL-4 mRNA expression was restricted to subcapsular regions of the human thymus, whereas cells expressing IL-7 mRNA were distributed throughout the thymic tissue, with some enrichment in subcapsular and cortical regions. Epithelial cells of the outer layer of Hassall's corpuscle strongly expressed IL-7 mRNA. IL-7 but not IL-4 or IL-2, stimulated strong proliferative activity and cellular expansion of triple-negative thymocytes. All three induced the appearance of TCR-gamma delta + cells within 4 days of culture. In the presence of IL-4 or IL-2, 30 to 55% of viable cells were TCR-gamma delta + ...
Scientific Reports
The cyclic GMP-AMP synthase (cGAS)/stimulator of interferon genes (STING) pathway is a cytosolic ... more The cyclic GMP-AMP synthase (cGAS)/stimulator of interferon genes (STING) pathway is a cytosolic sensor of microbial and host-derived DNA and plays a key role in innate immunity. Activation of STING by cyclic dinucleotide (CDN) ligands in human monocytes induces a type I interferon response and production of pro-inflammatory cytokines associated with the induction of massive cell death. In this study we have re-evaluated the effect of signal strength of STING activation on the cytokine plasticity of human monocytes. CDN (2′3′c-GAMP) and non-CDN (diABZI, MSA-2) STING ligands in the range of EC50 concentrations (15 μM 2′3′c-GAMP, 100 nM diABZI, 25 μM MSA-2) induced IFN-β, IP-10, and large amounts of IL-1β and TNF-α, but no IL-10 or IL-19. Interestingly, LPS-induced production of IL-10 and IL-19 was abolished in the presence of diABZI or MSA-2, whereas IL-1β and TNF-α were not inhibited. Surprisingly, we observed that tenfold lower (MSA-2, i.e. 2.5 μM) or 100-fold lower (diABZI, i.e. 1...
F1000 - Post-publication peer review of the biomedical literature, 2016
Highlights d Gut epithelial butyrophilin-like 1 (Btnl1) shapes the local gd T cell compartment d ... more Highlights d Gut epithelial butyrophilin-like 1 (Btnl1) shapes the local gd T cell compartment d Other organ-specific epithelial Btnl genes select cognate gd cells in other sites d Btnl heteromers can specifically activate gd T cells with cognate T cell receptors d Human BTNL genes reveal a conserved biology of epithelial T cell regulation Authors
LaboratoriumsMedizin, 2000
Zusammenfassung: Die Durchflußzytometrie hat sich in den vergangenen Jahren von einer reinen Fors... more Zusammenfassung: Die Durchflußzytometrie hat sich in den vergangenen Jahren von einer reinen Forschungsmethode zu einem fest etablierten diagnostischen Verfahren entwickelt. Die verwendeten Protokolle und die Indikationen zur durchflußzytometrischen Diagnostik sind ebenso wie die technischen Ansätze und Datenanalyse in ständiger Entwicklung begriffen. Das Ziel des vorliegenden Positionspapiers ist es, anhand aktueller Literatur und moderner Protokolle und Techniken die diagnostische Wertigkeit durchflußzytometrischer Methoden kritisch zu beleuchten. Dabei werden Verfahren, die nachgewiesenermaßen einen Stellenwert in der Primärdiagnostik und Therapiekontrolle besitzen, ebenso dargestellt wie Verfahren, deren Bedeutung bei einem hohen diagnostischen Potential noch nicht ausreichend belegt ist. Die Autoren beabsichtigten nicht, mit der vorliegenden Übersicht Standards zu definieren, sondern wollen für die Weiterentwicklung der Durchflußzytometrie, aber auch für die Verständigung mit anderen interessierten Fachgesellschaften eine Diskussionsgrundlage bereitstellen.
MedChemComm, 2015
A novel fluorescent derivative of bisphosphonate zoledronate was synthesized and shown to have co... more A novel fluorescent derivative of bisphosphonate zoledronate was synthesized and shown to have comparable functional activity as native zoledronate.
Cancer research, 2000
Programmed cell death (apoptosis) is primarily mediated by Fas ligand (FasL; CD95L) and the Fas r... more Programmed cell death (apoptosis) is primarily mediated by Fas ligand (FasL; CD95L) and the Fas receptor (Fas; CD95). In this study, FasL was detected by immunohistochemical analysis in 85% of breast carcinomas and 14% of fibroadenomas randomly chosen, indicating that high expression of FasL might play a role in tumor pathology. FasL and Fas levels as well as FasL:Fas ratios were further ascertained in 215 human breast tumors, including 199 invasive ductal carcinomas, by real-time quantitative reverse transcription-PCR and compared with expression levels and ratios found in 25 normal human tissues, in 37 fibroadenomas, and in 5 normal breast tissues. Among breast carcinomas, high FasL mRNA expression seems to be positively correlated with histological grading (n ؍ 212; P < 0.0001). A ratio of FasL:Fas mRNA transcripts >1 is found to be significantly associated with decreased patient's disease-free survival (n ؍ 211; P < 0.03) and increased mortality (n ؍ 211; P ؍ 0.19). A FasL:Fas ratio >1 is related to tumor progression scored by histological grading (n ؍ 212; P < 0.02). The selection process leading to highly aggressive breast tumor variants might be enhanced by FasL-mediated tumor fratricide, eventually a possible target for novel therapeutic strategies.
PLoS ONE, 2014
Pancreatic ductal adenocarcinoma (PDAC) is characterized by an extensive stroma being also presen... more Pancreatic ductal adenocarcinoma (PDAC) is characterized by an extensive stroma being also present in chronic pancreatitis (CP). Using immunohistochemistry, the stroma of CP and PDAC was comprehensively analyzed and correlated with epithelial/carcinoma-related alterations and clinicopathological patient characteristics. While there were no significant differences between CP and PDAC regarding the distribution of CD3+ T cells and a-SMA+ fibroblasts, proportions of CD4+ and CD8+ T cells were significantly lower and numbers of CD25+(CD4+) and FoxP3+(CD4+) regulatory T cells were greater in PDAC compared with CP. Macrophages were more prevalent in CP, but localized more closely to carcinoma cells in PDAC, as were cd-T cells. Duct-related FoxP3 and L1CAM expression increased from CP to PDAC, while vimentin expression was similarly abundant in both diseases. Moreover, stromal and epithelial compartments of well-differentiated tumors and CPs shared considerable similarities, while moderately and poorly differentiated tumors significantly differed from CP tissues. Analysis of 27 parameters within each pancreatic disease revealed a significant correlation of i) CD4+ and FoxP3+CD4+ T cells with FoxP3 expression in PDAC cells, ii) a-SMA+ fibroblasts with L1CAM expression and proliferation in PDAC cells, iii) CD3 and CD8 expression with cd-TCR expression in both pancreatic diseases and iv) CD68+ and CD163+ macrophages with vimentin expression in PDAC cells. High expression of FoxP3, vimentin and L1CAM in PDAC cells as well as a tumor-related localization of macrophages each tended to correlate with higher tumor grade. Multivariate survival analysis revealed a younger age at time of surgery as a positive prognostic marker for PDAC patients with the most frequently operated disease stage T3N1M0. Overall this study identified several interrelationships between stroma and epithelial/carcinoma cells in PDACs but also in CP, which in light of previous experimental data strongly support the view that the inflammatory stroma contributes to malignancy-associated alterations already in precursor cells during CP.
The Journal of Infectious Diseases, 2002
It has been shown that human gd T cells expressing the Vd1 T cell receptor (TCR) respond to the s... more It has been shown that human gd T cells expressing the Vd1 T cell receptor (TCR) respond to the spirochete Borrelia burgdorferi. Lysates of 3 Borrelia genospecies triggered the proliferation of peripheral-blood mononuclear cells not only from patients with skin manifestations of Borrelia infection and from patients with Lyme arthritis but also from healthy donors. However, with the exception of 1 patient with Lyme arthritis, no selective expansion of Vd1expressing gd T cells was induced. In contrast, synovial-fluid mononuclear cells (SFMC) from 3 of 5 patients with Lyme arthritis responded with a selective outgrowth of Vd1 gd T cells. Vd1 gd T cell lines established from SFMC coexpressed various TCR Vg chains, although Vg8 was preferentially used. Thus, the responsiveness to Borrelia antigens is not a general property of Vd1-expressing gd T cells but is largely restricted to Vd1 gd T cells recruited into the inflamed tissue. A minor subset of T cells expresses a CD3-associated gd T cell receptor (TCR) instead of the conventional ab TCR. In contrast to ab T cells, the germline-encoded TCR repertoire of gd T cells is strikingly small. In humans, there are only 6 expressed Vg genes (Vg2-Vg5 and Vg8 in the VgI family and Vg9 in the VgII family) and a similarly small number of Vd genes . In the peripheral blood of healthy adults, most (up to 95%) of gd T cells express Vd2Vg9, whereas Vd1 in association with various Vg genes is preferentially used by intraepithelial intestinal gd T cells [1-3]. Human Vd2Vg9 gd T cells recognize small phosphorylated molecules ("phosphoantigens") produced by various microorganisms, including Mycobacterium tuberculosis [2], whereas Vd1-expressing gd T cells recognize stress-inducible major histocompatibility complex (MHC) class I-related antigens and certain microbial ligands, such as cytomegalovirus and Borrelia burgdorferi lysates . In view of the known reactivity of Lyme arthritis synovialfluid Vd1 gd T cells to Borrelia lysates, we investigated whether
The Journal of Immunology, 2008
Chronic lymphocytic leukemia (CLL) is characterized by a highly variable clinical course. The rol... more Chronic lymphocytic leukemia (CLL) is characterized by a highly variable clinical course. The role of an autologous tumor-specific immune control contributing to the variable length of survival in CLL is poorly understood. We investigated whether humoral immunity specific for the CLL-associated Ag oncofetal Ag/immature laminin receptor (OFA/iLR) has a prognostic value in CLL. Among sera of 67 untreated patients with CLL, 23 (34.3%) had detectable OFA/iLR Abs that were reactive for at least one specific OFA/iLR epitope. Patients with humoral responses compared with patients with nonreactive sera had a longer progression-free survival (p = 0.029). IgG subclass analyses showed a predominant IgG1 and IgG3 response. OFA/iLR Abs were capable of recognizing and selectively killing OFA/iLR-expressing CLL cells in complement-mediated and Ab-dependent cellular cytotoxi cityassays. In the analysis of 11 CLL patients after allogeneic hematopoetic stem cell transplantation, 8 showed high values ...
The Journal of Immunology, 2006
The oncofetal Ag immature laminin receptor (OFA-iLR) is a potential target molecule for immunothe... more The oncofetal Ag immature laminin receptor (OFA-iLR) is a potential target molecule for immunotherapeutic studies in several tumor entities, including hematological malignancies. In the present study, we characterize two HLA-A*0201-presented epitopes eliciting strong OFA-iLR peptide-specific human cytotoxic T cell (CTLs) responses in vitro. Both allogeneic HLA-A*0201-matched and autologous CTLs recognized and killed endogenously OFA-iLR-expressing tumor cell lines and primary malignant cells from patients with hemopoietic malignancies in an MHC-restricted fashion but spared nonmalignant hemopoietic cells. Spontaneous OFA-iLR peptide-specific T cell reactivity was detectable in a significant proportion of leukemia patients. Interestingly, in patients with chronic lymphocytic leukemia and multiple myeloma but not in those with acute myeloid leukemia, significant frequencies of OFA peptide-specific CTLs could be detected in an early stage of disease but disappeared in patients with pro...
The Journal of Immunology, 2000
Taking advantage of a PCR technique that allows amplification of all variable region genes with e... more Taking advantage of a PCR technique that allows amplification of all variable region genes with equal efficiency, we defined three novel waves of TCR δ-chain transcription during thymic ontogeny. The canonical DV101-D2-J2 rearrangement was confined to a narrow window from days 14 to 18 of gestation, indicating that the postulated two consecutive γδ precursor waves bearing this canonical DV101 rearrangement will coincide on day 16. Neonatal δ-chain transcripts used a second wave of diverse Vα gene segments that are exclusively located in the δ locus-proximal gene cluster of intermingled single members of different Vα subfamilies. In the adult, only expression of a clan of three homologous subfamilies, ADV7, DV104, and ADV17, persists. The members of the ADV7 subfamily are also scattered across the α locus, but their usage does not show the position-dependent bias of the other Vα-to-δ rearrangements.
The Journal of experimental medicine, 1994
Structural diversity of lymphocyte antigen receptors (the immunoglobulin [Ig] of B cells and the ... more Structural diversity of lymphocyte antigen receptors (the immunoglobulin [Ig] of B cells and the alpha/beta or gamma/delta T cell receptor [TCR] of T cells) is generated through somatic rearrangements of V, D, and J gene segments. Classically, these recombination events involve gene segments from the same Ig or TCR locus. However, occurrence of "trans" rearrangements between distinct loci has also been described, although in no instances was the surface expression of the corresponding protein under normal physiological conditions demonstrated. Here we show that hybrid TCR genes generated by trans rearrangement between V gamma and (D) J beta elements are translated into functional antigen receptor chains, paired with TCR alpha chains. Like classical alpha/beta T cells, cells expressing these hybrid TCR chains express either CD4 or CD8 coreceptors and are frequently alloreactive. These results have several implications in terms of T cell repertoire selection and relationship...
The Journal of experimental medicine, 1988
From 1 to 23% of fetal human spleen or thymus cells (from the 20th to 24th week of gestation) wer... more From 1 to 23% of fetal human spleen or thymus cells (from the 20th to 24th week of gestation) were found to display a previously unrecognized CD2-/CD3+ phenotype. IL-2-dependent, long-term clones of CD2-/3+ T cells did not react with a panel of anti-CD2 mAbs and did not form rosettes with sheep erythrocytes. These results show that (a) significant numbers of CD2-/3+ T cells are present in fetal human spleen and/or thymus; and (b) in contrast to the widely accepted view, expression of CD2 is not a prerequisite for the expression of the CD3 molecular complex on human T cells.
Journal of Biological Chemistry, 2000
Upon binding of their ligands, death receptors belonging to the tumor necrosis factor (TNF) recep... more Upon binding of their ligands, death receptors belonging to the tumor necrosis factor (TNF) receptor family initiate a signaling pathway leading to the activation of caspases and ultimately apoptosis. TNF, however, in parallel elicits survival signals, protecting many cell types from cell death that can only be induced by combined treatment with TNF and inhibitors of protein synthesis. Here, we report that in NIH3T3 cells, apoptosis in response TNF and cycloheximide is not inhibited by the broad spectrum caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (zVAD.fmk). Moreover, treatment with zVAD.fmk sensitizes the cells to the cytotoxic action of TNF. Sensitization was also achieved by overexpression of a dominant-negative mutant of Fas-associated death domain protein and, to a lesser extent, by specific inhibition of caspase-8. A similar, but weaker sensitization of zVAD.fmk to treatment with the TNF-related apoptosis-inducing ligand (TRAIL) or anti-CD95 antibody was demonstrated. The unexpected cell death in response to TNF and caspase inhibition occurs despite the activation of nuclear factor B and c-Jun N-terminal kinases. The mode of cell death shows several signs of apoptosis including DNA fragmentation, although activation of caspase-3 was excluded. TNF/ zVAD.fmk-induced cell death is preceded by an accumulation of cells in the G 2 /M phase of the cell cycle, indicating an important role of cell cycle progression. This hypothesis is further strengthened by the observation that arresting the cells in the G 1 phase of the cell cycle inhibited TNF/zVAD.fmk-induced cell death, whereas blocking them in the G 2 /M phase augmented it.
Differential Regulation of Activation-Induced Cell Death in Individual Human T Cell Clones
International Archives of Allergy and Immunology, 2000
Background: Restimulation of T lymphocytes via the TCR/CD3 complex can result in CD95/CD95L-depen... more Background: Restimulation of T lymphocytes via the TCR/CD3 complex can result in CD95/CD95L-dependent activation-induced cell death (AICD). Although the correlation of AICD sensitivity to the T helper 1 phenotype was confirmed in different studies, the underlying mechanism is still debated. Thus, it has been suggested that in Th2 cells, AICD resistance is controlled by a TCR-induced upregulation of the CD95-associated inhibitory phosphatase, FAP-1. We and others demonstrated that AICD resistance is associated with a reduced surface expression of CD95L upon restimulation. Methods: Utilizing RT-PCR, Western blotting and flow cytometry, we analyzed time-dependent changes in levels of CD95L mRNA, cytosolic protein and surface expression in five long-term human T cell clones and polarized helper populations. Results: We confirm that the inducible CD95L surface expression is lower or absent in all tested AICD-resistant clones as compared to sensitive cells. It is of interest that striking...
Journal of Immunology, Jun 1, 2006
The oncofetal Ag immature laminin receptor (OFA-iLR) is a potential target molecule for immunothe... more The oncofetal Ag immature laminin receptor (OFA-iLR) is a potential target molecule for immunotherapeutic studies in several tumor entities, including hematological malignancies. In the present study, we characterize two HLA-A*0201-presented epitopes eliciting strong OFA-iLR peptide-specific human cytotoxic T cell (CTLs) responses in vitro. Both allogeneic HLA-A*0201-matched and autologous CTLs recognized and killed endogenously OFA-iLR-expressing tumor cell lines and primary malignant cells from patients with hemopoietic malignancies in an MHC-restricted fashion but spared nonmalignant hemopoietic cells. Spontaneous OFA-iLR peptide-specific T cell reactivity was detectable in a significant proportion of leukemia patients. Interestingly, in patients with chronic lymphocytic leukemia and multiple myeloma but not in those with acute myeloid leukemia, significant frequencies of OFA peptide-specific CTLs could be detected in an early stage of disease but disappeared in patients with progressive disease. The identification of OFA-iLR-derived peptide epitopes provides a basis for tumor immunological studies and therapeutic vaccination strategies in patients with OFA-iLR-expressing malignancies.
Frontiers in Immunology, Sep 30, 2022
gamma delta (gd) T cells, tumor immunotherapy, butyrophilins, cytokine programming, gamma delta T... more gamma delta (gd) T cells, tumor immunotherapy, butyrophilins, cytokine programming, gamma delta TCR repertoire, cost-effective manufacturing of gamma delta t cell therapies, chimeric antigen receptor-engineered gamma delta T cells, phosphoantigens The discovery of the gd T-cell receptor (TCR) and its ability to confer potent cytotoxic activity in CD3+ cells some 35 years ago sparked the initial proliferation in research that garnered widespread interest in the biology and function of gd T cells. The identification of major human gd TCR clonotypes, their tissue distributions as well as dynamic changes throughout ontogeny and in disease states have contributed to the appreciation of human gd T cell diversity. Despite this, incomplete understanding of mechanisms underlying the complexity of various gd T cell subsets in homeostasis, inflammation and malignancy restricted the focus of most early studies to blood circulating Vg9Vd2 T cells which could be robustly activated and expanded ex vivo using aminobisphosphonates such as zoledronate compared with other gd T cell subsets for which activating ligands were then largely unknown. This galvanized attempts to harness the tumoricidal potential of Vg9Vd2 T cells in clinical trials. Although these cells exhibited highly promising safety profiles in patients, early trial data revealed suboptimal anti-tumor efficacy. Despite these setbacks, recent breakthroughs in deciphering the unique antigen (Ag) binding modes of gd TCR coupled with high dimensional analyses of tissue-and disease-specific gd T cell subsets at single cell resolution led to renewed excitement in the development of gd T cell-based therapeutics. This Research Topic has compiled a series of nine articles of which five review our hitherto understanding of the multifaceted nature of gd T cells and four report original research providing new insights into the molecular and cellular regulation of a diverse repertoire of gd T cells, paving the way for next generation gd T cell-based tumor immunotherapies.
Frontiers in Immunology, Nov 12, 2021
Vitamin C (VitC), in addition to its role as a general antioxidant, has long been considered to p... more Vitamin C (VitC), in addition to its role as a general antioxidant, has long been considered to possess direct anti-cancer activity at high doses. VitC acts through oxidant and epigenetic mechanisms, which at high doses can exert direct killing of tumor cells in vitro and delay tumor growth in vivo. Recently, it has also been shown that pharmacologicdose VitC can contribute to control of tumors by modulating the immune system, and studies have been done interrogating the role of physiologic-dose VitC on novel adoptive cellular therapies (ACTs). In this review, we discuss the effects of VitC on anti-tumor immune cells, as well as the mechanisms underlying those effects. We address important unanswered questions concerning both VitC and ACTs, and outline challenges and opportunities facing the use of VitC in the clinical setting as an adjunct to immune-based anti-cancer therapies. Keywords: immune checkpoint therapy (ICT), CAR (chimeric antigen receptor) T cells, vitamin C (ascorbic acid), cancer immunotherapies, cancer biology
The Journal of Immunology
We have investigated the frequency and specificity of gamma delta+ cytotoxic lymphocyte precursor... more We have investigated the frequency and specificity of gamma delta+ cytotoxic lymphocyte precursors (CLP) under limiting dilution culture conditions. E rosette separated total T cells and CD3+CD4-CD8-TCR alpha beta- double-negative (DN) T cells were cocultured with allogeneic or autologous PBMC stimulator cells, and frequencies of alloreactive and autoreactive CLP were determined after 12 to 14 days against Con A blast target cells. Freshly isolated DN cells consisting of 82.3 +/- 8.2% gamma delta+ T cells did not exert cytolytic activity against K562 or anti-TCR gamma delta mAb-producing hybridoma cells. In striking contrast to E+ cells, the vast majority of alloantigen-stimulated clonally developing DN CLP did not show specificity for stimulator-derived target cells. Thus, frequencies of alloreactive and autoreactive CLP after alloantigenic stimulation were in the range of 1/100 to 1/4800 and 1/450 to 1/5000, respectively. After coculture with autologous stimulator cells, frequenci...
The Journal of Immunology
It is well established that IL-4 and IL-7 control the differentiation of TCR-gamma delta-expressi... more It is well established that IL-4 and IL-7 control the differentiation of TCR-gamma delta-expressing cells from CD3/TCR-negative thymic precursors. In this study, we have compared the in situ expression of IL-4 and IL-7 mRNA in human postnatal thymus with the in vitro effect of IL-4 and IL-7 on the expansion of TCR-gamma delta + cells from highly purified CD3-4-8- triple-negative thymocytes. IL-4 mRNA expression was restricted to subcapsular regions of the human thymus, whereas cells expressing IL-7 mRNA were distributed throughout the thymic tissue, with some enrichment in subcapsular and cortical regions. Epithelial cells of the outer layer of Hassall's corpuscle strongly expressed IL-7 mRNA. IL-7 but not IL-4 or IL-2, stimulated strong proliferative activity and cellular expansion of triple-negative thymocytes. All three induced the appearance of TCR-gamma delta + cells within 4 days of culture. In the presence of IL-4 or IL-2, 30 to 55% of viable cells were TCR-gamma delta + ...
Scientific Reports
The cyclic GMP-AMP synthase (cGAS)/stimulator of interferon genes (STING) pathway is a cytosolic ... more The cyclic GMP-AMP synthase (cGAS)/stimulator of interferon genes (STING) pathway is a cytosolic sensor of microbial and host-derived DNA and plays a key role in innate immunity. Activation of STING by cyclic dinucleotide (CDN) ligands in human monocytes induces a type I interferon response and production of pro-inflammatory cytokines associated with the induction of massive cell death. In this study we have re-evaluated the effect of signal strength of STING activation on the cytokine plasticity of human monocytes. CDN (2′3′c-GAMP) and non-CDN (diABZI, MSA-2) STING ligands in the range of EC50 concentrations (15 μM 2′3′c-GAMP, 100 nM diABZI, 25 μM MSA-2) induced IFN-β, IP-10, and large amounts of IL-1β and TNF-α, but no IL-10 or IL-19. Interestingly, LPS-induced production of IL-10 and IL-19 was abolished in the presence of diABZI or MSA-2, whereas IL-1β and TNF-α were not inhibited. Surprisingly, we observed that tenfold lower (MSA-2, i.e. 2.5 μM) or 100-fold lower (diABZI, i.e. 1...
F1000 - Post-publication peer review of the biomedical literature, 2016
Highlights d Gut epithelial butyrophilin-like 1 (Btnl1) shapes the local gd T cell compartment d ... more Highlights d Gut epithelial butyrophilin-like 1 (Btnl1) shapes the local gd T cell compartment d Other organ-specific epithelial Btnl genes select cognate gd cells in other sites d Btnl heteromers can specifically activate gd T cells with cognate T cell receptors d Human BTNL genes reveal a conserved biology of epithelial T cell regulation Authors
LaboratoriumsMedizin, 2000
Zusammenfassung: Die Durchflußzytometrie hat sich in den vergangenen Jahren von einer reinen Fors... more Zusammenfassung: Die Durchflußzytometrie hat sich in den vergangenen Jahren von einer reinen Forschungsmethode zu einem fest etablierten diagnostischen Verfahren entwickelt. Die verwendeten Protokolle und die Indikationen zur durchflußzytometrischen Diagnostik sind ebenso wie die technischen Ansätze und Datenanalyse in ständiger Entwicklung begriffen. Das Ziel des vorliegenden Positionspapiers ist es, anhand aktueller Literatur und moderner Protokolle und Techniken die diagnostische Wertigkeit durchflußzytometrischer Methoden kritisch zu beleuchten. Dabei werden Verfahren, die nachgewiesenermaßen einen Stellenwert in der Primärdiagnostik und Therapiekontrolle besitzen, ebenso dargestellt wie Verfahren, deren Bedeutung bei einem hohen diagnostischen Potential noch nicht ausreichend belegt ist. Die Autoren beabsichtigten nicht, mit der vorliegenden Übersicht Standards zu definieren, sondern wollen für die Weiterentwicklung der Durchflußzytometrie, aber auch für die Verständigung mit anderen interessierten Fachgesellschaften eine Diskussionsgrundlage bereitstellen.
MedChemComm, 2015
A novel fluorescent derivative of bisphosphonate zoledronate was synthesized and shown to have co... more A novel fluorescent derivative of bisphosphonate zoledronate was synthesized and shown to have comparable functional activity as native zoledronate.
Cancer research, 2000
Programmed cell death (apoptosis) is primarily mediated by Fas ligand (FasL; CD95L) and the Fas r... more Programmed cell death (apoptosis) is primarily mediated by Fas ligand (FasL; CD95L) and the Fas receptor (Fas; CD95). In this study, FasL was detected by immunohistochemical analysis in 85% of breast carcinomas and 14% of fibroadenomas randomly chosen, indicating that high expression of FasL might play a role in tumor pathology. FasL and Fas levels as well as FasL:Fas ratios were further ascertained in 215 human breast tumors, including 199 invasive ductal carcinomas, by real-time quantitative reverse transcription-PCR and compared with expression levels and ratios found in 25 normal human tissues, in 37 fibroadenomas, and in 5 normal breast tissues. Among breast carcinomas, high FasL mRNA expression seems to be positively correlated with histological grading (n ؍ 212; P < 0.0001). A ratio of FasL:Fas mRNA transcripts >1 is found to be significantly associated with decreased patient's disease-free survival (n ؍ 211; P < 0.03) and increased mortality (n ؍ 211; P ؍ 0.19). A FasL:Fas ratio >1 is related to tumor progression scored by histological grading (n ؍ 212; P < 0.02). The selection process leading to highly aggressive breast tumor variants might be enhanced by FasL-mediated tumor fratricide, eventually a possible target for novel therapeutic strategies.
PLoS ONE, 2014
Pancreatic ductal adenocarcinoma (PDAC) is characterized by an extensive stroma being also presen... more Pancreatic ductal adenocarcinoma (PDAC) is characterized by an extensive stroma being also present in chronic pancreatitis (CP). Using immunohistochemistry, the stroma of CP and PDAC was comprehensively analyzed and correlated with epithelial/carcinoma-related alterations and clinicopathological patient characteristics. While there were no significant differences between CP and PDAC regarding the distribution of CD3+ T cells and a-SMA+ fibroblasts, proportions of CD4+ and CD8+ T cells were significantly lower and numbers of CD25+(CD4+) and FoxP3+(CD4+) regulatory T cells were greater in PDAC compared with CP. Macrophages were more prevalent in CP, but localized more closely to carcinoma cells in PDAC, as were cd-T cells. Duct-related FoxP3 and L1CAM expression increased from CP to PDAC, while vimentin expression was similarly abundant in both diseases. Moreover, stromal and epithelial compartments of well-differentiated tumors and CPs shared considerable similarities, while moderately and poorly differentiated tumors significantly differed from CP tissues. Analysis of 27 parameters within each pancreatic disease revealed a significant correlation of i) CD4+ and FoxP3+CD4+ T cells with FoxP3 expression in PDAC cells, ii) a-SMA+ fibroblasts with L1CAM expression and proliferation in PDAC cells, iii) CD3 and CD8 expression with cd-TCR expression in both pancreatic diseases and iv) CD68+ and CD163+ macrophages with vimentin expression in PDAC cells. High expression of FoxP3, vimentin and L1CAM in PDAC cells as well as a tumor-related localization of macrophages each tended to correlate with higher tumor grade. Multivariate survival analysis revealed a younger age at time of surgery as a positive prognostic marker for PDAC patients with the most frequently operated disease stage T3N1M0. Overall this study identified several interrelationships between stroma and epithelial/carcinoma cells in PDACs but also in CP, which in light of previous experimental data strongly support the view that the inflammatory stroma contributes to malignancy-associated alterations already in precursor cells during CP.
The Journal of Infectious Diseases, 2002
It has been shown that human gd T cells expressing the Vd1 T cell receptor (TCR) respond to the s... more It has been shown that human gd T cells expressing the Vd1 T cell receptor (TCR) respond to the spirochete Borrelia burgdorferi. Lysates of 3 Borrelia genospecies triggered the proliferation of peripheral-blood mononuclear cells not only from patients with skin manifestations of Borrelia infection and from patients with Lyme arthritis but also from healthy donors. However, with the exception of 1 patient with Lyme arthritis, no selective expansion of Vd1expressing gd T cells was induced. In contrast, synovial-fluid mononuclear cells (SFMC) from 3 of 5 patients with Lyme arthritis responded with a selective outgrowth of Vd1 gd T cells. Vd1 gd T cell lines established from SFMC coexpressed various TCR Vg chains, although Vg8 was preferentially used. Thus, the responsiveness to Borrelia antigens is not a general property of Vd1-expressing gd T cells but is largely restricted to Vd1 gd T cells recruited into the inflamed tissue. A minor subset of T cells expresses a CD3-associated gd T cell receptor (TCR) instead of the conventional ab TCR. In contrast to ab T cells, the germline-encoded TCR repertoire of gd T cells is strikingly small. In humans, there are only 6 expressed Vg genes (Vg2-Vg5 and Vg8 in the VgI family and Vg9 in the VgII family) and a similarly small number of Vd genes . In the peripheral blood of healthy adults, most (up to 95%) of gd T cells express Vd2Vg9, whereas Vd1 in association with various Vg genes is preferentially used by intraepithelial intestinal gd T cells [1-3]. Human Vd2Vg9 gd T cells recognize small phosphorylated molecules ("phosphoantigens") produced by various microorganisms, including Mycobacterium tuberculosis [2], whereas Vd1-expressing gd T cells recognize stress-inducible major histocompatibility complex (MHC) class I-related antigens and certain microbial ligands, such as cytomegalovirus and Borrelia burgdorferi lysates . In view of the known reactivity of Lyme arthritis synovialfluid Vd1 gd T cells to Borrelia lysates, we investigated whether
The Journal of Immunology, 2008
Chronic lymphocytic leukemia (CLL) is characterized by a highly variable clinical course. The rol... more Chronic lymphocytic leukemia (CLL) is characterized by a highly variable clinical course. The role of an autologous tumor-specific immune control contributing to the variable length of survival in CLL is poorly understood. We investigated whether humoral immunity specific for the CLL-associated Ag oncofetal Ag/immature laminin receptor (OFA/iLR) has a prognostic value in CLL. Among sera of 67 untreated patients with CLL, 23 (34.3%) had detectable OFA/iLR Abs that were reactive for at least one specific OFA/iLR epitope. Patients with humoral responses compared with patients with nonreactive sera had a longer progression-free survival (p = 0.029). IgG subclass analyses showed a predominant IgG1 and IgG3 response. OFA/iLR Abs were capable of recognizing and selectively killing OFA/iLR-expressing CLL cells in complement-mediated and Ab-dependent cellular cytotoxi cityassays. In the analysis of 11 CLL patients after allogeneic hematopoetic stem cell transplantation, 8 showed high values ...
The Journal of Immunology, 2006
The oncofetal Ag immature laminin receptor (OFA-iLR) is a potential target molecule for immunothe... more The oncofetal Ag immature laminin receptor (OFA-iLR) is a potential target molecule for immunotherapeutic studies in several tumor entities, including hematological malignancies. In the present study, we characterize two HLA-A*0201-presented epitopes eliciting strong OFA-iLR peptide-specific human cytotoxic T cell (CTLs) responses in vitro. Both allogeneic HLA-A*0201-matched and autologous CTLs recognized and killed endogenously OFA-iLR-expressing tumor cell lines and primary malignant cells from patients with hemopoietic malignancies in an MHC-restricted fashion but spared nonmalignant hemopoietic cells. Spontaneous OFA-iLR peptide-specific T cell reactivity was detectable in a significant proportion of leukemia patients. Interestingly, in patients with chronic lymphocytic leukemia and multiple myeloma but not in those with acute myeloid leukemia, significant frequencies of OFA peptide-specific CTLs could be detected in an early stage of disease but disappeared in patients with pro...
The Journal of Immunology, 2000
Taking advantage of a PCR technique that allows amplification of all variable region genes with e... more Taking advantage of a PCR technique that allows amplification of all variable region genes with equal efficiency, we defined three novel waves of TCR δ-chain transcription during thymic ontogeny. The canonical DV101-D2-J2 rearrangement was confined to a narrow window from days 14 to 18 of gestation, indicating that the postulated two consecutive γδ precursor waves bearing this canonical DV101 rearrangement will coincide on day 16. Neonatal δ-chain transcripts used a second wave of diverse Vα gene segments that are exclusively located in the δ locus-proximal gene cluster of intermingled single members of different Vα subfamilies. In the adult, only expression of a clan of three homologous subfamilies, ADV7, DV104, and ADV17, persists. The members of the ADV7 subfamily are also scattered across the α locus, but their usage does not show the position-dependent bias of the other Vα-to-δ rearrangements.
The Journal of experimental medicine, 1994
Structural diversity of lymphocyte antigen receptors (the immunoglobulin [Ig] of B cells and the ... more Structural diversity of lymphocyte antigen receptors (the immunoglobulin [Ig] of B cells and the alpha/beta or gamma/delta T cell receptor [TCR] of T cells) is generated through somatic rearrangements of V, D, and J gene segments. Classically, these recombination events involve gene segments from the same Ig or TCR locus. However, occurrence of "trans" rearrangements between distinct loci has also been described, although in no instances was the surface expression of the corresponding protein under normal physiological conditions demonstrated. Here we show that hybrid TCR genes generated by trans rearrangement between V gamma and (D) J beta elements are translated into functional antigen receptor chains, paired with TCR alpha chains. Like classical alpha/beta T cells, cells expressing these hybrid TCR chains express either CD4 or CD8 coreceptors and are frequently alloreactive. These results have several implications in terms of T cell repertoire selection and relationship...
The Journal of experimental medicine, 1988
From 1 to 23% of fetal human spleen or thymus cells (from the 20th to 24th week of gestation) wer... more From 1 to 23% of fetal human spleen or thymus cells (from the 20th to 24th week of gestation) were found to display a previously unrecognized CD2-/CD3+ phenotype. IL-2-dependent, long-term clones of CD2-/3+ T cells did not react with a panel of anti-CD2 mAbs and did not form rosettes with sheep erythrocytes. These results show that (a) significant numbers of CD2-/3+ T cells are present in fetal human spleen and/or thymus; and (b) in contrast to the widely accepted view, expression of CD2 is not a prerequisite for the expression of the CD3 molecular complex on human T cells.
Journal of Biological Chemistry, 2000
Upon binding of their ligands, death receptors belonging to the tumor necrosis factor (TNF) recep... more Upon binding of their ligands, death receptors belonging to the tumor necrosis factor (TNF) receptor family initiate a signaling pathway leading to the activation of caspases and ultimately apoptosis. TNF, however, in parallel elicits survival signals, protecting many cell types from cell death that can only be induced by combined treatment with TNF and inhibitors of protein synthesis. Here, we report that in NIH3T3 cells, apoptosis in response TNF and cycloheximide is not inhibited by the broad spectrum caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (zVAD.fmk). Moreover, treatment with zVAD.fmk sensitizes the cells to the cytotoxic action of TNF. Sensitization was also achieved by overexpression of a dominant-negative mutant of Fas-associated death domain protein and, to a lesser extent, by specific inhibition of caspase-8. A similar, but weaker sensitization of zVAD.fmk to treatment with the TNF-related apoptosis-inducing ligand (TRAIL) or anti-CD95 antibody was demonstrated. The unexpected cell death in response to TNF and caspase inhibition occurs despite the activation of nuclear factor B and c-Jun N-terminal kinases. The mode of cell death shows several signs of apoptosis including DNA fragmentation, although activation of caspase-3 was excluded. TNF/ zVAD.fmk-induced cell death is preceded by an accumulation of cells in the G 2 /M phase of the cell cycle, indicating an important role of cell cycle progression. This hypothesis is further strengthened by the observation that arresting the cells in the G 1 phase of the cell cycle inhibited TNF/zVAD.fmk-induced cell death, whereas blocking them in the G 2 /M phase augmented it.
Differential Regulation of Activation-Induced Cell Death in Individual Human T Cell Clones
International Archives of Allergy and Immunology, 2000
Background: Restimulation of T lymphocytes via the TCR/CD3 complex can result in CD95/CD95L-depen... more Background: Restimulation of T lymphocytes via the TCR/CD3 complex can result in CD95/CD95L-dependent activation-induced cell death (AICD). Although the correlation of AICD sensitivity to the T helper 1 phenotype was confirmed in different studies, the underlying mechanism is still debated. Thus, it has been suggested that in Th2 cells, AICD resistance is controlled by a TCR-induced upregulation of the CD95-associated inhibitory phosphatase, FAP-1. We and others demonstrated that AICD resistance is associated with a reduced surface expression of CD95L upon restimulation. Methods: Utilizing RT-PCR, Western blotting and flow cytometry, we analyzed time-dependent changes in levels of CD95L mRNA, cytosolic protein and surface expression in five long-term human T cell clones and polarized helper populations. Results: We confirm that the inducible CD95L surface expression is lower or absent in all tested AICD-resistant clones as compared to sensitive cells. It is of interest that striking...