ABSTRACT
Natural killer (NK) cell subpopulations from 8 HLA-matched but killer cell immunoglobulin-like receptor (KIR)/HLA-ligand-mismatched patient-donor pairs were analyzed in the course of allogeneic hematopoietic stem cell transplantation (HCT). The patients' post-transplantation NKG2A-/LIR-1- NK cells, which expressed only inhibitory KIRs for which the patient had no HLA class I ligands, showed higher cytotoxic capacity than the NKG2A-/LIR-1- NK cells lacking any inhibitory KIRs that remained tolerant throughout the course of HCT. The NKG2A+ NK cell subpopulations displayed the highest levels of cytotoxic activation, which appeared to be significantly enhanced in comparison with that in allogeneic graft's donors. LIR-1- NK cells were much more frequent after HCT than LIR-1+ NK cells and LIR-1 expression on NKG2A+ or NKG2A- NK cells was associated with significantly lower cytotoxic activities. Thus NKG2A-/LIR-1- NK cells expressing only HLA-mismatched KIRs show a partial break in tolerance in the first year following HCT. The failure to exclude LIR-1+ cells within the NKG2A- NK cell subset in previous studies could explain the earlier conflicting results. Thus systemic immune activation in patients following HCT augments the GvL effect through both increasing overall NK cell activities and partially breaking tolerance of unlicensed NK cells.
Subject(s)
HLA Antigens/immunology , Hematopoietic Stem Cell Transplantation/methods , Killer Cells, Natural/immunology , Receptors, Immunologic/immunology , Adult , Aged , Graft vs Leukemia Effect , Humans , Male , Middle Aged , NK Cell Lectin-Like Receptor Subfamily C , Receptors, KIR/immunologyABSTRACT
Myelodysplastic syndromes (MDS) comprise a spectrum of clonal stem cell disorders which are currently defined according to the classification scheme of the revised 2008 WHO classification but which may be further refined in the future. The clinical presentation is often characterized by unexplained isolated or multiple peripheral blood cytopenias resulting in anemia, bleeding events or increased susceptibility to infections. The generally hypercellular, but rarely hypocellular and occasionally fibrotic bone marrow shows dysplastic features in ≥ 10 % of cells of at least one of the hematopoietic lineages. These features and enhanced apoptosis, stem cell senescence and immunologic dysregulation result in ineffective hematopoiesis. Diagnostics in MDS relies on complementary consideration of hematological, morphological and cytogenetic/molecular parameters. Methods include marrow and peripheral blood cytology, cytogenetics, fluorescence in situ hybridization (FISH), trephine bone marrow biopsy examination, immunophenotyping and the evaluation of molecular markers by established and new techniques. Mutations affecting growth factor receptors, cell cycle and apoptosis regulators, intracellular signaling, transcription factors, epigenetic regulation and the splicosome are involved in MDS pathogenesis and progression.
Subject(s)
Myelodysplastic Syndromes/genetics , Myelodysplastic Syndromes/pathology , Biomarkers, Tumor/genetics , Biopsy, Needle , Blood Cells/pathology , Bone Marrow/pathology , Cross-Sectional Studies , Cytogenetic Analysis/methods , DNA Mutational Analysis/methods , Hematopoietic Stem Cells/pathology , Humans , Immunophenotyping/methods , In Situ Hybridization, Fluorescence/methods , Myelodysplastic Syndromes/classification , Myelodysplastic Syndromes/epidemiology , Prognosis , Risk FactorsSubject(s)
Chromosome Aberrations , Chromosomes, Human, Pair 8/genetics , Histiocytosis, Langerhans-Cell/diagnosis , Histiocytosis, Langerhans-Cell/genetics , Sarcoma, Myeloid/diagnosis , Sarcoma, Myeloid/genetics , Trisomy/genetics , Diagnosis, Differential , Female , Humans , Middle Aged , PrognosisABSTRACT
Acute graft-versus-host disease (GvHD) limits the applicability of allogeneic hematopoietic cell transplantation for the treatment of leukemia. GvHD occurs as a consequence of multiple activating events in antigen-presenting cells (APCs) and T cells (Tcs). Spleen tyrosine kinase (Syk) is an intracellular non-receptor tyrosine kinase involved in multiple signaling events of immune cells. Therefore, we hypothesized that Syk may be a promising target to inhibit GvHD, which involves activation of different immune cell populations. In vivo expansion of luciferase(+) donor Tcs in mice developing GvHD was reduced by treatment with the Syk inhibitor Fostamatinib, which led to increased survival and reduced histologically confirmed GvHD severity. Importantly, in vivo and in vitro cytotoxicity against leukemia target cells and anti-murine cytomegalovirus immune responses were not impacted by Fostamatinib. In APCs Syk inhibition reduced the expression of costimulatory molecules and disrupted cytoskeletal organization with consecutive APC migratory defects in vitro and in vivo while phagocytic activity remained intact. On the basis of these immunomodulatory effects on different cell populations, we conclude that Syk targeting in alloantigen-activated Tcs and APCs with pharmacologic inhibitors, already applied successfully in anti-lymphoma therapy, has clinical potential to reduce GvHD, especially as anti-leukemia and anti-viral immunity were preserved.
Subject(s)
Antigen-Presenting Cells/immunology , Graft vs Host Disease/prevention & control , Intracellular Signaling Peptides and Proteins/antagonists & inhibitors , Leukemia/therapy , Oxazines/therapeutic use , Protein-Tyrosine Kinases/antagonists & inhibitors , Pyridines/therapeutic use , Spleen/enzymology , Aminopyridines , Animals , Blotting, Western , Bone Marrow Transplantation , Cell Movement , Cell Proliferation , Cytomegalovirus , Cytomegalovirus Infections/immunology , Cytomegalovirus Infections/virology , Dendritic Cells/immunology , Female , Flow Cytometry , Graft vs Host Disease/enzymology , Graft vs Host Disease/etiology , Graft vs Leukemia Effect , Intracellular Signaling Peptides and Proteins/metabolism , Leukemia/immunology , Lymphocyte Activation , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Morpholines , Phosphorylation , Protein-Tyrosine Kinases/metabolism , Pyrimidines , Syk Kinase , T-Lymphocytes/immunology , Transplantation, HomologousABSTRACT
BACKGROUND: The aim of this study was to investigate the prognostic effect of tumour-infiltrating lymphocytes (TILs) in serous stage III ovarian carcinoma to determine TIL clonality and to correlate this to Her2/neu expression. METHODS: Formalin-fixed and paraffin-embedded ovarian carcinomas were examined for CD20-, CD3-, CD4- and CD8-positive lymphocytes (n=100), and for Her2/neu-positive tumour cells (n=55/100) by immunohistochemistry. Clonality analysis was carried out by T-cell receptor gamma (TCRgamma) gene rearrangements (n=93/100). Statistical analyses included experimental and clinico-pathological variables, as well as disease-free (DFS) and overall (OS) survival. RESULTS: CD20-positive B lymphocytes were present in 57.7% (stromal)/33.0% (intraepithelial) and CD3-positive T lymphocytes in 99.0% (stromal)/90.2% (intraepithelial) of ovarian carcinomas. Intraepithelial CD3-positive T lymphocytes were correlated with improved DFS in optimally debulked patients (P=0.0402). Intraepithelial CD8-positive T lymphocytes were correlated with improved OS in all optimally debulked patients (P=0.0201) and in those undergoing paclitaxel/carboplatin therapy (P=0.0092). Finally, rarified and clonal TCRgamma gene rearrangements were detected in 37 out of 93 (39.8%) and 15 out of 93 (16.1%) cases, respectively. This was marginally associated with improved DFS (P=0.0873). Despite a significant correlation of HER2/neu status and intraepithelial CD8-positive lymphocytes (P=0.0264), this was non-directional (R=-0.257; P=0.0626). CONCLUSION: Improved survival of ovarian cancer patients is related to the infiltration, clonal selection and intraepithelial persistence of T lymphocytes.
Subject(s)
CD8-Positive T-Lymphocytes/immunology , Lymphocytes, Tumor-Infiltrating/immunology , Ovarian Neoplasms/immunology , Ovarian Neoplasms/mortality , Adult , Aged , Aged, 80 and over , Female , Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor , Humans , Middle Aged , Neoplasm Staging , Ovarian Neoplasms/pathology , Receptor, ErbB-2/analysisABSTRACT
BACKGROUND: Chronic neutrophilic leukaemia (CNL) is a rare myeloproliferative disorder of elderly patients characterised by sustained neutrophilia and splenomegaly. The diagnosis of CNL requires the exclusion of BCR/ABL positive chronic myelogenous leukaemia (CML) and of leukaemoid reactions (LRs). The differentiation between CNL and LR is problematic because both conditions share similar morphological features; it is also important because patients with CNL generally have a poor prognosis. AIMS: To determine whether CNL and LR could be distinguished on the basis of different clonality patterns. METHODS: Blood samples from 52 women were studied using the human androgen receptor gene assay (HUMARA). RESULTS: Monoclonality was found in the neutrophils in all 17 patients with different myeloproliferative syndromes (MPSs), including those with CNL. In four of the patients with CNL, autologous T cells were also monoclonal, suggesting that they belonged to the neoplastic clone. This finding was in contrast to other MPSs in which T cells were almost always polyclonal. Of nine patients with clinically suspected LR, the neutrophils of five were polyclonal, whereas three patients had monoclonal neutrophils, suggesting that they might be in the process of developing an MPS. Among 26 healthy blood donors, 20 had polyclonal neutrophils and five showed skewed clonality patterns. One case of LR and one normal blood donor were scored "not informative" at the HUMARA locus. CONCLUSIONS: Clonality studies of blood neutrophils using HUMARA aid in distinguishing female patients with monoclonal CNL from those with LR. For the diagnosis of CNL, monoclonality of the neutrophils should be demonstrated whenever possible.
Subject(s)
Leukemia, Neutrophilic, Chronic/diagnosis , Leukemoid Reaction/diagnosis , Neoplastic Stem Cells/pathology , Adult , Aged , Aged, 80 and over , Diagnosis, Differential , Dosage Compensation, Genetic , Female , Humans , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/diagnosis , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics , Leukemia, Neutrophilic, Chronic/genetics , Leukemoid Reaction/genetics , Middle Aged , Neutrophils/pathology , Receptors, Androgen/genetics , Receptors, Androgen/metabolismABSTRACT
There is growing interest in the use of innate immune reactions in the therapy and prophylaxis of various diseases. Natural T (NT) lymphocytes that recognize infected cells or microbial compounds without the classical genetic restriction by polymorphic MHC molecules are crucial components of innate immunity. NT cells bearing the Vgamma9Vdelta2 T-cell receptor (TCR) are broadly reactive against intracellular pathogens, can lyse human immunodeficiency virus (HIV) infected cells, and release cytokines capable of regulating HIV replication. The potent antiviral activities of Vgamma9Vdelta2 T cells may help to contain viral spread during acute HIV infection and/or to prevent the establishment of viral persistence. Substantial changes in the composition and function of circulating gammadelta T-cell pools occur in HIV-infected patients. These changes a) may contribute to the etiopathogenesis of opportunistic infections and neoplasms, and b) are partly reversed by highly active anti-retroviral therapy (HAART). In addition to direct antiviral activities, activated gammadelta T cells influence dendritic cell maturation and the adaptive alphabeta T-cell response. Vgamma9Vdelta2 T cells can be stimulated in vivo and in vitro by various nonpeptidic antigens (NpAgs) and recent animal experimental data suggest that activated Vgamma9Vdelta2 T cells may help to control SIV replication. Currently, NpAgs are being assessed as potential therapeutic agents in AIDS, tuberculosis and certain cancers susceptible to Vgamma9Vdelta2 T-cell effector mechanisms.
Subject(s)
HIV Infections/immunology , HIV Infections/virology , Receptors, Antigen, T-Cell, gamma-delta/metabolism , Receptors, Antigen, T-Cell, gamma-delta/physiology , T-Lymphocytes/immunology , T-Lymphocytes/virology , Acquired Immunodeficiency Syndrome/therapy , Animals , Antiretroviral Therapy, Highly Active , Antiviral Agents/pharmacology , B-Lymphocytes/virology , Cell Differentiation , Cytokines/metabolism , Humans , Killer Cells, Natural/virology , Ligands , Models, Biological , Polymorphism, GeneticABSTRACT
Chronic neutrophilic leukaemia (CNL) is a rare BCR/ABL negative myeloproliferative disorder of elderly patients, showing sustained neutrophilia and splenomegaly. Differentiation between CNL and leukaemoid reactions (LR) is problematic since both conditions share similar morphological features but is essential because CNL patients generally have a poor prognosis. We studied blood samples from 10 female patients with CNL or LR using the HUMARA assay to determine clonality patterns in neutrophils. T-lymphocytes of the patients were investigated as an internal control cell population. In all five CNL patients the neutrophils, and in four of them also T-lymphocytes were monoclonal, indicating that the latter may also originate from the neoplastic clone. In LR patients the neutrophils and T-lymphocytes were generally polyclonal except in one patient showing monoclonal neutrophils suggesting that this patient might be in the process of developing a myeloproliferative disorder. In females clonality studies of blood neutrophils using HUMARA aid in distinguishing patients with monoclonal CNL from polyclonal LR.
Subject(s)
Leukemia, Neutrophilic, Chronic/pathology , Adult , Aged , Aged, 80 and over , Diagnosis, Differential , Follow-Up Studies , Humans , Middle Aged , Neutrophils/pathology , Platelet Count , T-Lymphocytes/pathology , Treatment OutcomeABSTRACT
In 1986 we diagnosed chronic idiopathic myelofibrosis (CIMF) in a 45-year-old asymptomatic patient with hepatosplenomegaly. In 1996 splenectomy was performed because of hypersplenism, and chemotherapy with hydroxyurea was initiated. In 1999 generalised lymphadenopathy of chronic lymphocytic leukaemia (B-CLL) developed. A trephine biopsy showed leukaemic bone marrow infiltration. On heteroduplex analysis we found a clonal rearrangement of IgH in the leukaemic lymphocytes. The coincidence of chronic myeloproliferative and lymphoproliferative diseases in the same patient is a rare phenomenon. According to the relevant literature, seven cases with a combination of CIMF/CLL have been reported. Possible pathomechanisms for the development of such coincidences are: 1) a bilineage manifestation of a pluripotent stem cell proliferation, 2) independent proliferations of two distinct cell lines under a common leukaemogenic stimulus or 3) an accidental association. These coincidence cases often showed a mild clinical course and also in our case, the patient is still alive and in a stable disease condition 16 years after the initial diagnosis.
Subject(s)
Leukemia, Lymphocytic, Chronic, B-Cell/complications , Leukemia, Lymphocytic, Chronic, B-Cell/pathology , Primary Myelofibrosis/complications , Primary Myelofibrosis/pathology , Diagnosis, Differential , Humans , Male , Middle AgedABSTRACT
Local immunosuppression may explain the failure of an effective immune response against solid tumors. Although it is well known that the interstitial pH is significantly lower in solid tumors than in normal tissue, only a few studies in the mouse system have investigated the influence of this acidic milieu on the anti-tumoral cytotoxic response. Here the authors report the suppression of human non-major histocompatibility complex (MHC)-restricted cytotoxicity against tumor cells by an acidic extracellular pH (pHe). Unstimulated peripheral blood mononuclear cells, lymphokine-activated killer (LAK) cells, and natural killer cell clones were used as effector cells. According to pH measurements in solid tumors, representative pH values of 7.2 to 5.3 were chosen during the cytotoxic assays. Target cell lysis was measured using two nonradioactive fluorometric methods, namely two-color flow cytometry and a modified calcein-release assay, which allowed cell-mediated cytotoxicity to be measured and compared with that in adherent targets. Using K562, Daudi, or Raji as suspended target cell lines, the cytotoxic activity of unstimulated peripheral blood mononuclear cells and of LAK cells was markedly reduced by a decreasing pHe. An extracellular pH of 5.8 to 5.3 resulted in a nearly complete loss of the cytotoxic response. This pHe-dependent impairment of the killing activity could also be shown for killer cells stimulated with interleukins-7 and -12, phytohemagglutinin, or lipopolysaccharide. The lytic potential of homogeneous natural killer cell clones as effectors was also strictly influenced by the surrounding pH. The pHe dependence of the non-MHC-restricted killer cell functions against tumor cells seems to be a general phenomenon, because the cytolytic activity of LAK cells against six human adherent tumor cell lines (HeLa, HepG2, LS174T, LS174Te, MCF-7, and RT112) was also clearly reduced under acidic conditions. To initiate the killing process, adhesion molecules play an important role in recognition and binding of the target cell. However, flow cytometric analysis revealed that the expression pattern of relevant adhesion molecules was unaffected by acidic pHe. In conclusion, these data clearly indicate an inhibition of non-MHC-restricted cytotoxicity against tumor cells by an acidic pHe, which may contribute to the failure of immunosurveillance against solid tumors. Consequently, efforts to enhance the anti-tumoral cytotoxicity by immunotherapies may have limited success.
Subject(s)
Cytotoxicity, Immunologic , Hydrogen-Ion Concentration , Immunosuppression Therapy , Major Histocompatibility Complex/immunology , Neoplasms/immunology , Neoplasms/therapy , T-Lymphocytes, Cytotoxic/immunology , Acids , HeLa Cells , Humans , K562 Cells , Killer Cells, Lymphokine-Activated/immunology , Killer Cells, Lymphokine-Activated/metabolism , Lymphocyte Activation/immunology , Lymphocyte Subsets/immunology , Lymphocyte Subsets/metabolism , T-Lymphocytes, Cytotoxic/metabolism , Tumor Cells, CulturedSubject(s)
Antigens, Neoplasm/immunology , Autoimmunity/immunology , Receptors, Antigen, T-Cell, gamma-delta/immunology , T-Lymphocytes/immunology , Antigen Presentation/immunology , Antigens, Bacterial/immunology , Burkitt Lymphoma/immunology , Histocompatibility Antigens Class I/immunology , Humans , T-Lymphocytes, Cytotoxic/immunologyABSTRACT
Because of their hypervariable regions and somatic mutations, the antigen receptor molecules of lymphomas (idiotypes) are tumor-specific antigens and attractive targets for antilymphoma immunotherapy. For the optimal induction of human idiotype-specific cytotoxic T cells (CTL), idiotype was presented to CD8(+) peripheral blood mononuclear cells by monocyte-derived autologous dendritic cells (DC) after the endocytosis of idiotype protein or by idiotype-expressing DC. Recombinant idiotype was obtained as a functionally folded Fab fragment by periplasmic expression in Escherichia coli. Idiotype-expressing DC were generated by transduction with recombinant Semliki forest virus vectors encompassing heavy- or light-chain idiotype genes. Autologous lymphoblastoid cell lines stably transfected with Epstein-Barr virus-based idiotype expression vectors were used as target cells to detect idiotype-specific lysis. CTL stimulated with idiotype-loaded DC showed strong specific, CD8-mediated, and major histocompatibility complex (MHC) class I-restricted cytotoxicity against autologous heavy- and light-chain idiotype. In contrast, stimulation with idiotype-transduced DC resulted in only moderate natural killer cell activity. These data confirm the existence of idiotype-specific CTL in patients with lymphoma, define a "good manufacturing practice"-compatible protocol for the generation of these cells without the requirement of viable lymphoma cells, and favor the processing of exogenous antigen over DC transduction for the induction of MHC I-restricted CTL against idiotypes with unknown antigenicity. (Blood. 2000;95:1342-1349)
Subject(s)
CD8-Positive T-Lymphocytes/immunology , Dendritic Cells/immunology , Immunoglobulin Idiotypes/immunology , Leukemia, Lymphocytic, Chronic, B-Cell/immunology , Lymphoma, Follicular/immunology , T-Lymphocytes, Cytotoxic/immunology , Adult , Amino Acid Sequence , Antibodies, Viral/immunology , Base Sequence , Cytotoxicity, Immunologic , DNA Primers , Female , Humans , Immunoglobulin Heavy Chains/genetics , Immunoglobulin Heavy Chains/immunology , Immunoglobulin Idiotypes/chemistry , Immunoglobulin Idiotypes/genetics , Immunoglobulin Light Chains/genetics , Immunoglobulin Light Chains/immunology , Leukemia, Lymphocytic, Chronic, B-Cell/genetics , Leukemia, Lymphocytic, Chronic, B-Cell/pathology , Lymphoma, Follicular/genetics , Lymphoma, Follicular/pathology , Male , Middle Aged , Molecular Sequence Data , Monocytes/immunology , Polymerase Chain Reaction , Recombinant Proteins/chemistry , Recombinant Proteins/immunology , Semliki forest virus/immunologySubject(s)
Red-Cell Aplasia, Pure/etiology , Anticonvulsants/adverse effects , Antitubercular Agents/adverse effects , Autoimmune Diseases/immunology , Azathioprine/adverse effects , Bone Marrow Cells/pathology , Fanconi Anemia/etiology , Female , Humans , Immunosuppressive Agents/adverse effects , Infant , Isoniazid/adverse effects , Killer Cells, Natural/immunology , Leukemia, Lymphoid/etiology , Male , Parvovirus B19, Human , Phenytoin/adverse effects , Pregnancy , Pregnancy Complications, Hematologic , Red-Cell Aplasia, Pure/immunology , Red-Cell Aplasia, Pure/pathology , T-Lymphocytes/immunologyABSTRACT
An expression system for rapid and standardized production of human recombinant immunoglobulin Fab fragments in E. coli was developed. Functional folding of the Fab fragments was accomplished by the dicistronic expression vector pFab.gammakappa containing specialized leader sequences to direct the immunoglobulin heavy and light chains to the periplasmic bacterial space. A C-terminal hexahistidine tag of the Fd chain facilitated metal affinity chromatography and purification to homogeneity as assessed by SDS PAGE and silver staining. Specific antigen recognition by a hybridoma-derived Fab fragment was indistinguishable from that of the corresponding monoclonal antibody. This protocol may be useful for analysis of the antigen specificity of human B cells and for convenient production of lymphoma-derived idiotype protein for vaccination strategies. To obtain unmodified immunoglobulin cDNA sequences from small human biopsies for insertion into pFab.gammakappa, oligo(dG)-tailed cDNA was amplified with an oligo(dC)- and nested mu or kappa constant region-specific primers. Using single sets of primers for each class of immunoglobulin transcripts, the products of this anchored PCR reflected the relative abundance of the starting cell population and permitted reliable identification of clonal, lymphoma-derived sequences for subsequent expression cloning.
Subject(s)
Cancer Vaccines/immunology , Immunoglobulin Fab Fragments/biosynthesis , Lymphoma, B-Cell/immunology , Recombinant Proteins/biosynthesis , Cloning, Molecular , Epitopes , Escherichia coli/genetics , Genes, Immunoglobulin , Humans , Immunoglobulin Fab Fragments/genetics , Immunoglobulin Fab Fragments/immunology , Polymerase Chain Reaction , Recombinant Proteins/immunology , Tumor Cells, Cultured , VaccinationSubject(s)
Bone Marrow Transplantation/adverse effects , Mycobacterium Infections/blood , Mycobacterium Infections/complications , Mycobacterium bovis , Receptors, Antigen, T-Cell, gamma-delta/analysis , Severe Combined Immunodeficiency/etiology , Humans , Infant , Male , Severe Combined Immunodeficiency/pathology , T-Lymphocyte Subsets/chemistryABSTRACT
OBJECTIVE: To determine the feasibility of a transport IVF program involving air transportation of oocytes. DESIGN: Prospective cohort study. SETTING: Regional hospital (Hôpital de Chicoutimi) and University Infertility Center (McGill Reproductive Center, Montreal). PATIENT(S): The first series of patients referred for IVF or IVF and ICSI, for a variety of indications, who opted for inclusion in the transport IVF program. INTERVENTION(S): The IVF-ET with ovarian stimulation and oocyte collection at the peripheral unit and transport of the oocytes by airplane to the McGill Reproductive Center where IVF or ICSI was performed. MAIN OUTCOME MEASURE(S): Clinical pregnancy. RESULT(S): Seven couples, in the first series, underwent nine cycles of transport IVF treatment. Two also underwent ICSI. There were two clinical pregnancies. CONCLUSION(S): Transport IVF using air travel is possible and opens the possibility for this type of program to be implemented in large countries with scattered populations, such as the United States, Canada, and Australia.
Subject(s)
Aircraft , Fertilization in Vitro/methods , Microinjections , Specimen Handling , Adult , Buserelin/administration & dosage , Cohort Studies , Embryo Transfer , Estradiol/blood , Female , Humans , Infertility/therapy , Male , Ovulation Induction , Pregnancy , Prospective Studies , TwinsABSTRACT
OBJECTIVES: The objectives of this study were to assess cytokine secretion in human renal cell carcinoma (RCC) and to identify cytokines contributing to the immunomodulatory effect of tumor cells. METHODS: Cytokine secretion in the supernatant of primary tumor cell cultures (PTCC) and corresponding cell lines (CL) was assayed using ELISA. Tumor cells were characterized by morphology, immunocytochemistry, and flow-cytometric analysis. Tumor-cell-induced T cell activation was determined by coculture of gamma delta and alpha beta T cell clones with tumor CL. RESULTS: We assessed the cytokine secretion of tumor cells from 27 PTCC and their corresponding CL (3/27) of RCC. We found that RCC predominantly produced both pro-inflammatory and T-cell-inhibitory cytokines, such as IL-8, IL-6, GM-CSF, TNF-alpha, IL-10 and TGF-beta 1. CL were adapted to serum-free medium which may prove as a useful tool in future studies of cytokine secretion in RCC. In addition, we used gamma delta and alpha beta T cell clones to assess the immunomodulatory effect of tumor cells from RCC and found that predominantly gamma delta T cells were activated by RCC. CONCLUSIONS: Our data suggest that RCC produce large amounts of both pro-inflammatory and T-cell-inhibitory cytokines that potentially could influence the immune response of the host, especially tumor-specific cytotoxic T cells.