RESUMEN
BACKGROUND: We have previously established tumour T-cell lines, both from the skin and from the blood of patients with a cutaneous T-cell lymphoma (CTCL). In one patient, the tumour cells and the derived cell lines had a CD3+ CD4+ CD8- phenotype and a trisomy of chromosome 7. They expressed three T-cell receptor (TCR) beta-chain transcripts, but only one was productively rearranged and expressed at the cell membrane. OBJECTIVES: In the present study, we tried to isolate a fast-growing new tumour T-cell line from the same patient. PATIENTS/METHODS: We performed direct cell cloning of the skin tumour lymphocyte population, which led to the isolation of an interleukin-2-dependent highly proliferative T-cell subclone, named Cou-L3, with a CD3+ TCR-Vbeta13+ CD4- CD8alphaalpha+ phenotype. RESULTS: We demonstrated that Cou-L3 was identical to the original clonal tumour CD3+ Vbeta13+ CD4+ CD8- cells, as it expressed the same rearranged TCR-Vbeta13 chain. We further studied the functional activity of these CD8alphaalpha+ Vbeta13+ Cou-L3 cells. We found that these cells exhibited CD3-redirected cytotoxic activity. CONCLUSIONS: An immunophenotypic shift, with a change from a CD4+ to a CD8+ phenotype, has been already reported in association with disease progression in CTCL. However, in these cases, there has been no demonstration that the phenotypic change involved the same T-cell clone. The present study is the first report of the phenotypic heterogeneity of the tumour clonal cell population in CTCL.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Linfoma Cutáneo de Células T/patología , Neoplasias Cutáneas/patología , Células Tumorales Cultivadas , Anciano , Anciano de 80 o más Años , Complejo CD3/análisis , Humanos , Técnicas para Inmunoenzimas , Inmunofenotipificación , Linfoma Cutáneo de Células T/inmunología , Masculino , Micosis Fungoide/patología , Células Madre Neoplásicas/inmunología , Células Madre Neoplásicas/patología , Receptores de Antígenos de Linfocitos T/metabolismo , Neoplasias Cutáneas/inmunología , Subgrupos de Linfocitos T/inmunologíaRESUMEN
Primary cutaneous T-cell lymphomas (CTCL) are a heterogeneous group of malignant mature T-cell proliferations most often presenting as mycosis fungoides (MF) or its leukemic variant, Sezary syndrome (SS). No specific cell surface markers are presently available to distinguish the circulating malignant clone from normal lymphocytes. Using the previously established CTCL cell lines, Cou-LS and Pno, we have detected two leucocyte cell surface antigens with aberrant expression on CTCL cells. The NK-receptor (NKR) p140/KIR3DL2 normally expressed by NK and CD8+ T-cells was detected on the surface of CTCL cell lines as well as on freshly isolated CD4+PBL from SS patients. Further on, p140 marked in situ SS cells, distinguishing them from p140-negative tumor cells of patch plaque MF. SC5 is a newly described activation-related intracellular inhibitory receptor expressed on the surface of a minor PBL subset. We found that SC5 expression was significantly increased in SS cells and correlated to p140 expression. Moreover, cross-linking of SC5 molecules inhibited the malignant cell proliferation induced by anti-CD3 mAbs. The identification of these new structures on circulating SS tumor cells seems to be important both for the understanding of CTCL pathophysiology and for the clinical management of SS patients.
Asunto(s)
Linfoma Cutáneo de Células T/metabolismo , Receptores de Superficie Celular/análisis , Receptores Inmunológicos/análisis , Neoplasias Cutáneas/metabolismo , Antígenos de Superficie/análisis , Biomarcadores de Tumor/análisis , Linfocitos T CD4-Positivos/química , Humanos , Linfoma Cutáneo de Células T/diagnóstico , Linfoma Cutáneo de Células T/terapia , Receptores KIR , Receptores KIR3DL2 , Síndrome de Sézary/metabolismo , Neoplasias Cutáneas/diagnóstico , Neoplasias Cutáneas/terapia , Células Tumorales CultivadasRESUMEN
We have established several tumor T cell lines, both from the skin and from the blood of a patient with an MHC class II-/class I+, CD4+ cutaneous T cell lymphoma (CTCL). These cell lines, like the initial tumor cells, had a CD3+CD4+CD8- phenotype. We also isolated two cytotoxic T lymphocyte clones from the tumor site of this CTCL patient. These clones displayed a CD4+CD8dim+ (TC5) and CD4+CD8- (TC7) phenotype and mediated a specific MHC class I-restricted cytotoxic activity toward noncultured tumor cells and autologous tumor cell lines. Despite surface expression of Fas on tumor cells and Fas-L induction on TC5 and TC7 cell membrane after coculture with autologous tumor cells, the CD4+ CTL clones did not use this cytotoxic mechanism to lyse their specific target. TC7 used a granzyme/perforin-dependent pathway, whereas TC5 used a TRAIL-dependent mechanism. Quantitative analysis of cytokine mRNA expression indicated that while the tumor cells displayed a Th2-type profile, the CTL clones expressed Th1-type cytokines. Preincubation of TIL clones with autologous tumor cells in a short-term culture induced their activation and subsequent amplification of the Th1-type response, which indicates a direct contribution of the malignant cells in the Th1/Th2 imbalance. However, we found that tumor cells produced high amounts of TGF-beta, which could explain the inhibition of a specific antitumor immune response. Another mechanism to avoid the host immune response was the expression of CD158a, CD158b, p70, and CD94/NKG2A inhibitory receptors by tumor-specific lymphocytes. Finally, we present recent data on new antigen structures expressed both by long-term CTCL lines and uncultured tumor cells.
Asunto(s)
Linfocitos Infiltrantes de Tumor/inmunología , Linfoma Cutáneo de Células T/inmunología , Neoplasias Cutáneas/inmunología , Linfocitos T Citotóxicos/inmunología , Escape del Tumor , Antígenos de Neoplasias/metabolismo , Línea Celular , Células Clonales , Citocinas/biosíntesis , Humanos , Inmunofenotipificación , Receptores de Superficie Celular/metabolismo , Receptores Inmunológicos/metabolismo , Receptores KIR , Receptores KIR2DL1 , Receptores KIR2DL3 , Subgrupos de Linfocitos T/clasificación , Subgrupos de Linfocitos T/inmunología , Factor de Crecimiento Transformador beta/biosíntesisRESUMEN
Cutaneous T cell lymphomas are a clonal proliferation of CD4+ T lymphocytes primarily involving the skin. Mycosis fungoides is an epidermotropic CD4+ cutaneous T cell lymphoma, and a more aggressive form, Sezary syndrome, occurs when the malignant cells become nonepidermotropic. The role of neuropeptides in the growth and chemotaxis capacity of cutaneous T cell lymphoma cells remains unknown. In this report, we found that cutaneous T cell lymphoma cells, similarly to normal resting or activated peripheral lymphocytes, were able to bind neurotensin. We used an interleukin-2-dependent cutaneous T cell lymphoma malignant T cell line derived from cutaneous T cell lymphoma lesions in order to study the role of neurotensin in the proliferation and migration of these malignant cells. First, we determined that the malignant cells expressed neurotensin receptors on their cell membrane. Functional results indicated that neurotensin did not stimulate the growth of the cell line. In contrast, this neuropeptide inhibited the proliferation of the tumor cells in response to exogenous interleukin-2. Furthermore, we found that neurotensin enhanced both spontaneous and chemoattractant-induced migration of the malignant cells. This suggests that neurotensin in skin can play a role in the disease by locally limiting the growth of the cutaneous T cell lymphoma tumor cells in response to cytokines and by enhancing their chemotaxis capacity.
Asunto(s)
Linfoma Cutáneo de Células T/metabolismo , Receptores de Neurotensina/metabolismo , Neoplasias Cutáneas/metabolismo , Linfocitos T CD4-Positivos/metabolismo , Linfocitos T CD4-Positivos/patología , Citometría de Flujo , Humanos , Linfoma Cutáneo de Células T/patología , Neoplasias Cutáneas/patología , Células Tumorales CultivadasRESUMEN
Natural cytotoxic effector functions are regulated by a multitude of opposing signals provided by immunoglobulin and lectin-like functional molecules. While inhibitory receptors possess immunoreceptor tyrosine-based inhibition motif (ITIM) cytoplasmic sequences recruiting tyrosine phosphatases, activatory receptors require association with accesory immunoreceptor tyrosine-based activation motif (ITAM)-bearing molecules. One considerable group of natural cytotoxic cell receptors are specific for classical and non-classical class I antigens and detect both qualitative and quantitative changes in the autologous MHC-I phenotype. Non-MHC-I-specific receptors provide signaling in the absence of MHC-I antigens or in response to not well-known stress-induced antigens. NK cell receptors may equally participate in the regulation of target cell functions through contact or souble mediator-dependent mechanisms. The identification of NK cell regulating molecules has led to the elucidation of more general principles underlying immune homeostasis.
Asunto(s)
Citotoxicidad Inmunológica , Receptores Inmunológicos/metabolismo , Animales , Humanos , Inmunoglobulinas/metabolismo , Células Asesinas Naturales/inmunología , Lectinas/metabolismo , Receptores KIRRESUMEN
Using a newly generated monoclonal antibody we identified the 96 kDa transmembrane receptor SC5 expressed simultaneously on a human Sezary cell line and a minor T cell subset in normal individuals. SC5 antigen was detected mostly on CD45RO+ lymphocytes from both CD4+ and CD8+ subsets as well as on natural killer and B lineage cells. SC5 surface expression increased very early after polyclonal stimulation of CD3+ cells due to the transfer of intracellular SC5 molecules to the cell membrane. Engagement of SC5 receptor by its monoclonal antibody inhibited the anti-CD3-induced proliferation and cytokine secretion of peripheral blood T cells and cell clones, whereas SC5 monoclonal antibody did not affect the cytotoxic activity of CD8+ T cell clones. Extensive phenotypic analysis revealed that the percentage of SC5+ CD4+ circulating lymphocytes in Sezary syndrome patients was significantly increased in comparison with controls (p < 0.01) and correlated with the morphologically detected percentage of Sezary syndrome cells in peripheral blood (p < 0.001). In one patient we clearly demonstrated that the circulating malignant T cells coexpress SC5 molecules. Importantly, ligation of SC5 receptor in a cutaneous T cell lymphoma cell line profoundly inhibited the anti-CD3-induced proliferation. Consequently, the expression of SC5 receptor in the peripheral blood of Sezary syndrome patients may serve not only to detect the presence of circulating malignant CD4+ cells but also as a target for immunotherapy.
Asunto(s)
Linfoma de Células T/metabolismo , Receptores de Superficie Celular/metabolismo , Neoplasias Cutáneas/metabolismo , Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/farmacología , Células Sanguíneas/metabolismo , Complejo CD3/inmunología , División Celular/efectos de los fármacos , Células Cultivadas , Humanos , Linfocitos/patología , Linfoma de Células T/inmunología , Linfoma de Células T/patología , Receptores de Superficie Celular/química , Receptores de Superficie Celular/inmunología , Síndrome de Sézary/metabolismo , Neoplasias Cutáneas/inmunología , Neoplasias Cutáneas/patologíaRESUMEN
Interleukin (IL) -8 is a neutrophil chemoattractant cytokine with proinflammatory and growth-promoting activities, which is involved in the pathogenesis of several inflammatory diseases. It is found in high amounts in lesional biopsies of pustular diseases such as psoriasis and palmoplantar pustulosis. We report a 50-year-old woman with a 10-year history of erythroderma with disseminated pustulosis. Skin biopsies showed an epidermotropic infiltrate composed of atypical CD4+ CD8+ lymphocytes with numerous admixed neutrophils. Peripheral blood flow cytometric analysis revealed a major clonal subset of CD3+ CD4+ CD8+ T-cell receptor Vbeta22+ atypical lymphocytes. Bone marrow biopsy, lymph node biopsy and computed thoracoabdominal tomography were normal. Serologies for human T-cell lymphotropic virus type I and human immunodeficiency virus were negative. Our patient's status deteriorated despite topical (nitrogen mustard, psoralen plus ultraviolet A) and systemic (interferon, methotrexate, multiagent chemotherapy) treatments, and she finally died. We showed that our patient's peripheral blood lymphocytes (PBL) spontaneously produced high amounts of IL-8. In contrast, PBL of patients with classical Sézary syndrome produced lower amounts of IL-8. The production of IL-8 by tumour T cells could explain this unusual clinical and histopathological presentation of cutaneous T-cell lymphoma as disseminated pustulosis.
Asunto(s)
Dermatitis Exfoliativa/inmunología , Interleucina-8/biosíntesis , Linfoma Cutáneo de Células T/inmunología , Proteínas de Neoplasias/biosíntesis , Neoplasias Cutáneas/inmunología , Dermatitis Exfoliativa/patología , Resultado Fatal , Femenino , Humanos , Linfoma Cutáneo de Células T/patología , Persona de Mediana Edad , Neoplasias Cutáneas/patologíaRESUMEN
Vaccine-based therapies are being developed for a variety of cancers and their efficacy will be determined by their ability to stimulate T cells in the secondary lymphoid tissue. We found that T cells isolated from human secondary lymphoid organs (LT-T), in contrast to peripheral blood T cells (PB-T) are hyporesponsive to cross-linked anti-CD3 mAb (CD3c) even in the presence of exogenous IL-2. Using mAb to trigger CD2 and CD28 co-stimulatory molecules, we found that such dual co-stimulation of LT-T induces profound and sustained responses including CD25 expression, IL-2 secretion and proliferation. Different levels of co-stimulation produced a hierarchical pattern of responses in LT-T, which correlated with the degree of CD3-TCR down-regulation. Mature antigen-presenting cells (APC) restored the capacity of LT-T to proliferate to stimulation of the CD3-TCR complex. Blocking studies demonstrated that optimal proliferation was critically dependent on co-stimulation via CD2 and CD28 engaged by their ligands on the APC. Therefore, LT-T have increased co-stimulatory requirements as compared to PB-T, i.e. multiple co-stimulatory signals coupled to CD3-TCR triggering. Furthermore, LT-T were found to be dependent on APC for survival, in contrast to PB-T. Clearly, LT-T do not behave in a comparable way to PB-T and in vitro experiments assessing novel cancer vaccines should therefore use LT-T as the most appropriate population of responder T cells.
Asunto(s)
Ganglios Linfáticos/inmunología , Linfocitos T/inmunología , Adulto , Anticuerpos Monoclonales/farmacología , Células Presentadoras de Antígenos/inmunología , Antígenos CD2/inmunología , Antígenos CD28/inmunología , Complejo CD3/inmunología , Supervivencia Celular , Células Cultivadas , Niño , Regulación hacia Abajo , Humanos , Interleucina-2/farmacología , Leucocitos Mononucleares/inmunología , Ganglios Linfáticos/citología , Activación de Linfocitos , Linfoma no Hodgkin/inmunología , Complejo Receptor-CD3 del Antígeno de Linfocito T/inmunología , Receptores de Interleucina-2/inmunología , Bazo/citología , Bazo/inmunologíaRESUMEN
CD100 is the first semaphorin described in lymphoid tissues, where it has been shown to be associated with a serine kinase activity. Semaphorins are molecules involved in axon pathfinding during nerve development and act as repellent guidance cues. In the nervous system semaphorins exist as either membrane-bound or secreted forms. We report here a spontaneous processing of membrane CD100, suggesting that it is also produced as a diffusable semaphorin from lymphoid cells. Monomeric and homodimeric forms of CD100 are expressed by T lymphocytes and CD100-transfected fibroblasts. We demonstrate that CD100 is released through a proteolytic process blocked by metalloprotease inhibitors. In T cells, only soluble CD100 dimers are produced, suggesting that CD100 dimerization is required for proteolysis. In agreement, we observe that increasing membrane dimers strongly favors shedding of the molecule. By expressing a CD100 molecule mutated at cysteine 674 into a COS cell system, we additionally demonstrate that this particular residue in the extracellular domain of the molecule is required for dimerization. Finally, we show that staurosporine, a serine kinase inhibitor, enhances the membrane cleavage of CD100. Together these results demonstrate that membrane CD100 is cleaved by a metalloprotease-dependent process, which is probably regulated by phosphorylation. Mainly, these findings shed light on a possible function for the semaphorin region of CD100 as a long range guidance cue in the immune system.
Asunto(s)
Antígenos CD , Endopeptidasas/metabolismo , Espacio Extracelular/inmunología , Glicoproteínas de Membrana/metabolismo , Semaforinas , Linfocitos T/metabolismo , Células 3T3 , Adyuvantes Inmunológicos/farmacología , Animales , Células COS , Membrana Celular/enzimología , Membrana Celular/inmunología , Membrana Celular/metabolismo , Cisteína/genética , Cisteína/fisiología , Dimerización , Ácido Edético/farmacología , Ácido Egtácico/farmacología , Inhibidores Enzimáticos/farmacología , Espacio Extracelular/enzimología , Espacio Extracelular/metabolismo , Humanos , Hidrólisis/efectos de los fármacos , Yodoacetamida/farmacología , Células Jurkat , Glicoproteínas de Membrana/antagonistas & inhibidores , Glicoproteínas de Membrana/genética , Metaloendopeptidasas/antagonistas & inhibidores , Ratones , Mutagénesis Sitio-Dirigida , Estructura Terciaria de Proteína/genética , Solubilidad , Estaurosporina/farmacología , Linfocitos T/enzimología , Linfocitos T/inmunología , TransfecciónRESUMEN
CD100 is a human 150-kDa homodimer expressed at the surface of most hemopoietic cells, and its gene belongs to the Ig and semaphorin gene families. Semaphorin genes encode soluble and membrane-bound proteins, most of which have been shown to act as chemorepellents on growth cone guidance. CD100 is discrete, as it is a transmembrane leukocyte surface molecule that can also exist in a soluble form. While our previous studies using mAbs suggested that the transmembrane form of CD100 plays a role in lymphocyte activation, no function was shown for its soluble form. Here, we investigated the effect of soluble CD100 in a cell migration assay; both CD100 spontaneously shed from a stable transfectant and soluble recombinant CD100 inhibited spontaneous and chemokine-induced migration of human monocytes. Interestingly, only the dimeric form of CD100 exerted an effect. Moreover, soluble CD100 inhibited migration of cells from monocytic and B cell lineages. A similar inhibitory effect on migration was observed with H-SemaIII, but not H-SemaIV, semaphorins. In addition, both CD100 and H-SemaIII were recognized by two CD100 mAbs in an ELISA, and one of these mAb abolished the inhibitory effect of each of these semaphorins. We also provide evidence that CD100 and H-SemaIII act through the same receptor on immune cells, which is not neuropilin-1. Furthermore, we describe a function on immune cells for H-SemaIII, a semaphorin to date only studied in the nervous system.
Asunto(s)
Antígenos CD , Proteínas Portadoras/fisiología , Inhibición de Migración Celular , Movimiento Celular/inmunología , Glicoproteínas de Membrana/fisiología , Proteínas del Tejido Nervioso/fisiología , Semaforina-3A , Semaforinas , Células 3T3 , Animales , Anticuerpos Monoclonales/metabolismo , Anticuerpos Monoclonales/farmacología , Células COS , Proteínas Portadoras/metabolismo , Movimiento Celular/genética , Células Clonales/citología , Células Clonales/inmunología , Ensayo de Inmunoadsorción Enzimática , Humanos , Células Jurkat , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/inmunología , Glicoproteínas de Membrana/metabolismo , Ratones , Monocitos/citología , Monocitos/inmunología , Factores de Crecimiento Nervioso/farmacología , Proteínas del Tejido Nervioso/metabolismo , Neuropilina-1 , Receptores de Superficie Celular/metabolismo , Solubilidad , Linfocitos T/citología , Linfocitos T/inmunología , Transfección , Células U937/citología , Células U937/inmunologíaRESUMEN
Tumor cells of patients with cutaneous T-cell lymphoma (CTCL) have the cell surface phenotype of mature T-helper lymphocytes, and it may be impossible to differentiate them from nonmalignant lymphocytes in skin and blood. Until now, no specific cell membrane marker of CTCL has been reported. In the current study, it is reported for the first time that CTCL cells express the major histocompatibility complex class I binding p140-killer cell immunoglobulin-like receptor, which has been described on a minor subset of natural killer lymphocytes and on a marginal circulating CD8(+) T lymphocyte subset. Interestingly, the molecular characterization of this KIR expressed by CTCL allowed us to isolate a novel allelic form of p140-KIR3DL, resulting in 4 amino acid substitutions, 3 in the extracellular immunoglobulin-like domain of the protein and one in the cytoplasmic region. This finding is likely to be important both for the pathophysiology and for the clinical treatment of patients with CTCL.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Células Asesinas Naturales/inmunología , Linfoma Cutáneo de Células T/diagnóstico , Linfoma Cutáneo de Células T/inmunología , Receptores Inmunológicos/metabolismo , Secuencia de Aminoácidos , Anticuerpos Monoclonales , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/patología , Humanos , Linfoma Cutáneo de Células T/patología , Datos de Secuencia Molecular , Proteínas de Neoplasias/inmunología , Proteínas de Neoplasias/metabolismo , Pruebas de Precipitina , Isoformas de Proteínas/inmunología , Isoformas de Proteínas/metabolismo , Receptores Inmunológicos/inmunología , Receptores KIR , Alineación de Secuencia , Células Tumorales CultivadasRESUMEN
Inhibitory receptors on natural killer cells and on a minority of T lymphocytes are major histocompatibility complex class Ia or Ib specific. We have previously reported several tumor-specific cytotoxic T cell clones infiltrating a CD4(+) V beta 13(+) cutaneous T cell lymphoma. These clones mediated a specific major histocompatibility complex class I-restricted cytotoxic activity toward the uncultured tumor cells and autologous long-term tumor T cell lines. In this study, we cultured with interleukin-2 the peripheral blood lymphocytes of the same patient a few weeks before invasion of the blood by tumor cells. We report the rapid and selective expansion of a CD8(+) V beta 13(+) lymphoid population. This population was clonal, as it expressed a unique T cell receptor-V beta junctional region. V beta 13(+) tumor cells and V beta 13(+) reactive T cells were shown to have different junctional sequences. The CD8(+) reactive clone was functional, as it had a specific autologous tumor-specific, human leukocyte antigen-A2 restricted, cytotoxic activity. This clone coexpressed high levels of CD158a, CD158b, p70, and CD94/NKG2A inhibitory receptors. Interestingly, we found that anti-CD158a and anti-CD158b monoclonal antibodies could inhibit anti-CD3 redirected cytotoxicity mediated by the reactive clonal population. Further, an anti-human leukocyte antigen-B/C monoclonal antibody enhanced the specific cytotoxic activity of the clone against autologous tumor cells. These results are the first evidence that inhibitory receptor expression can lead to the inhibition of cutaneous T cell lymphoma-specific T cell responses.
Asunto(s)
Linfoma Cutáneo de Células T/metabolismo , Linfoma Cutáneo de Células T/patología , Receptores Inmunológicos/biosíntesis , Anciano , Anciano de 80 o más Años , Anticuerpos Monoclonales/farmacología , Linfocitos T CD8-positivos/inmunología , Células Clonales , Antígenos HLA-A/inmunología , Humanos , Células Asesinas Naturales/inmunología , Masculino , Receptores KIR , Receptores KIR2DL1 , Receptores KIR2DL3 , Linfocitos T Citotóxicos/citología , Linfocitos T Citotóxicos/inmunologíaRESUMEN
Since the CD101 molecule is expressed on a major subpopulation of HLA-DR(+), CD1a(+), CD1c(+) cutaneous dendritic cells (DC), we studied the functional role of CD101 on cutaneous DC. Anti-CD101 monoclonal antibody (mAb) inhibited the proliferation of T cells induced by cutaneous DC. There was a synergistic inhibition between anti-CD101 mAb and anti-CD86/anti-CD80 mAb. Anti-CD101 mAb exerted its inhibitory effect when binding to the CD101 expressed on cutaneous DC. No positive role of CD101 putative ligand expressed by T cells in T cell proliferation was demonstrated, as T cells proliferated in response to soluble anti-CD3 mAb in the presence of CD86-transfected cells but not in the presence of CD101-transfected cells. Of major significance is the fact that IL-10 was produced by cutaneous DC after CD101 triggering with anti-CD101 mAb, while IL-10 secretion was up-regulated in mixed cutaneous DC-T cell cultures after CD101 triggering. Furthermore, IL-10-neutralizing mAb could reverse the inhibition induced by anti-CD101 mAb. Our results demonstrate that the CD101 triggering on cutaneous DC inhibits T cell proliferation via IL-10 production, suggesting an important regulatory role played by the CD101 molecule on DC during T cell activation.
Asunto(s)
Comunicación Celular/inmunología , Células Dendríticas/inmunología , Interleucina-10/inmunología , Glicoproteínas de Membrana/inmunología , Linfocitos T/inmunología , Animales , Antígenos CD , División Celular/inmunología , Línea Celular , Humanos , Interleucina-10/biosíntesis , Activación de Linfocitos , RatonesRESUMEN
CDR3 of the functional rearranged T-cell receptor variable beta region (TCR-Vbeta) transcript was sequenced in order to demonstrate for the first time the identity between a long-term cultured T-cell line derived from a cutaneous T-cell lymphoma (CTCL) patient and the malignant T-cell clone present in the blood. The patient's peripheral blood lymphocyte-derived cultured T-cell line had a CD3(+)Vbeta22(+)CD4(+)CD8alphaalpha(+)CD25(-) phenotype. It was named Pno and had been cultured for more than 1 year. Both fresh and long-term-cultured tumor cells proliferated highly in response to interleukin-7 (IL-7), and exogeneous IL-7 prevented Pno lymphocytes from apoptosis and maintained high levels of Bcl-2 expression. This unique malignant cloned lymphocyte line was further used to carry out functional studies. The results indicated that the CD3/TCR structures expressed by the Pno lymphocytes were functional because an immobilized anti-CD3 monoclonal antibody (mAb) or the combination of a soluble anti-CD3 mAb with submitogenic doses of phorbol 12 beta-myristate 13 alpha-acetate induced a proliferative response. Further, the CD2 and CD28 coreceptors were functional because they were able to induce a strong proliferative response upon their specific stimulation. Finally, the Pno T cell line had a Th3-type cytokine profile because it produced high amounts of the immunosuppressor cytokine tumor growth factor-beta1 (TGF-beta1). This high production of TGF-beta1 may inhibit antitumor specific responses in CTCL.
Asunto(s)
Antígenos CD4/inmunología , Antígenos CD8/inmunología , Interleucina-7/inmunología , Linfoma Cutáneo de Células T/inmunología , Subgrupos de Linfocitos T/inmunología , Femenino , Humanos , Linfoma Cutáneo de Células T/patología , Persona de Mediana Edad , Subgrupos de Linfocitos T/patología , Células Tumorales CultivadasRESUMEN
We have previously described two cytotoxic T lymphocyte clones isolated from lymphocytes infiltrating a human major histocompatibility complex class II-/class I+, CD4+ cutaneous T cell lymphoma. These clones displayed a CD4+CD8dim+ (TC5) and CD4+ CD8- (TC7) phenotype and mediated a specific major histocompatibility complex class I-restricted cytotoxic activity toward Cou-LB autologous tumor cell line. Our studies were performed to elucidate the mechanism involved in T-cell-clone-mediated cytotoxicity and to determine the cytokine profile of both the lymphoma cell line and specific cytotoxic T lymphocyte clones. The results indicate that, despite surface expression of Fas receptor on Cou-LB and Fas ligand induction on TC5 and TC7 cell membranes, the CD4+ cytotoxic T lymphocyte clones do not use this cytotoxic mechanism to lyse their specific target. The TC7 clone uses instead a granzyme-perforin-dependent pathway. Furthermore, quantitative analysis of Th1 and Th2 cytokine mRNA expression in the cutaneous T cell lymphoma cell line as well as in TC5 and TC7 clones indicated that, whereas the tumor cells display a Th2-type profile (interleukin-4, interleukin-6, and interleukin-10), the cytotoxic T lymphocyte clones express Th1-type cytokines (interferon-gamma, granulocyte macrophage colony stimulating factor, and interleukin-2). In addition, preincubation of the tumor-infiltrating lymphocyte clones with autologous tumor cells induced their activation and subsequent amplification of the Th1-type response. These results indicate a direct contribution of the malignant cells in the Th1/Th2 imbalance observed frequently in cutaneous T cell lymphoma patients and suggest their potential role in depressed cell-mediated immunity. Identification of CD4+ Th1-type cytotoxic T lymphocyte clones, the tumor antigen they recognize, and optimization of their cytokine expression profile should be useful for the design of new immunotherapy protocols in cutaneous T cell lymphoma.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Citocinas/fisiología , Linfoma Cutáneo de Células T/patología , Neoplasias Cutáneas/patología , Linfocitos T Citotóxicos/citología , Células TH1/metabolismo , Receptor fas/metabolismo , Anciano , Anciano de 80 o más Años , Células Clonales , Factor Estimulante de Colonias de Granulocitos y Macrófagos/fisiología , Humanos , Inmunoterapia , Interferón gamma/fisiología , Interleucina-10/fisiología , Interleucina-2/fisiología , Interleucina-4/fisiología , Interleucina-6/fisiología , Masculino , Factor de Crecimiento Transformador beta/fisiología , Células Tumorales CultivadasRESUMEN
AIMS: Our objective was to study the expression of a recently identified cell surface molecule, CD101 and in Langerhans cell histiocytosis (LCH) patients as CD101 has been shown to be present on dendritic cells. We wanted to determine if CD101 expression could be helpful for the diagnosis of LCH in conjunction with other markers (CD1a, S100 protein), and could be predictive of the evolution and dissemination of the disease. METHODS AND RESULTS: The expression of CD101 was studied by immunohistochemical technique in 11 cases of Langerhans cell histiocytosis on frozen sections. The expression of CD101 was positive in nine cases, high in six cases and low in three cases. There was no expression in the other two cases. No correlation with the evolution, the localization or the dissemination of the disease could be evidenced. CONCLUSIONS: CD101 is a new phenotypic marker that might be useful in combination with other markers for the diagnosis of LCH. However, as the anti-CD101 antibody works only in frozen sections, its value is limited compared to anti-CD1a antibody.
Asunto(s)
Histiocitosis de Células de Langerhans/inmunología , Glicoproteínas de Membrana/inmunología , Presentación de Antígeno , Antígenos CD , Antígenos de Diferenciación de Linfocitos T/inmunología , Histiocitosis de Células de Langerhans/patología , Humanos , Inmunohistoquímica , Lactante , Recién Nacido , Glicoproteínas de Membrana/biosíntesisRESUMEN
Human CD100, the first semaphorin identified in the immune system, is a transmembrane protein involved in T-cell activation. In the present study, we showed that activation of peripheral blood or tonsillar B lymphocytes induced the expression of CD100 in CD38(+)CD138(-) cell populations, including in CD148(+) subpopulations, thus expressing a memory B-cell-like phenotype. Using an in vitro enzymatic assay, we found that protein tyrosine phosphatase (PTP) activities were immunoprecipitated with CD100 in these cell populations, which were isolated by cell sorting, as well as in most B-cell lines representing various stages of B-cell differentiation. Immunodepletion and Western blotting experiments demonstrated that CD45 was the PTP associated with CD100 in cell lines displaying pre-B, activated B, and pre-plasma cell phenotypes. CD45 also accounted for PTP activity immunoprecipitated with CD100 in CD38(+)CD138(-) cells sorted after activation of peripheral blood or tonsillar B lymphocytes. In contrast, no CD100-CD45 association was observed in plasma cell lines corresponding to the terminal B-cell differentiation stage. CD148, the other transmembrane PTP known to be implicated in lymphocyte signaling pathways, was either only partly involved in the CD100-associated PTP activity or not expressed in plasma cell lines, indicating the association of CD100 with another main PTP. Our data show that CD100 is differentially expressed and can functionally associate with distinct PTPs in B cells depending on their activation and maturation state. They also provide evidence for a switch in the CD100-associated PTP at terminal stage of B-cell differentiation.
Asunto(s)
Antígenos CD , Linfocitos B/metabolismo , Antígenos Comunes de Leucocito/metabolismo , Glicoproteínas de Membrana/biosíntesis , Proteínas Tirosina Fosfatasas/metabolismo , Semaforinas , ADP-Ribosil Ciclasa , ADP-Ribosil Ciclasa 1 , Antígenos de Diferenciación/análisis , Linfocitos B/citología , Diferenciación Celular , Línea Celular , Regulación del Desarrollo de la Expresión Génica , Humanos , Memoria Inmunológica , Inmunofenotipificación , Antígenos Comunes de Leucocito/genética , Antígenos Comunes de Leucocito/aislamiento & purificación , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/aislamiento & purificación , NAD+ Nucleosidasa/análisis , Tonsila Palatina/citología , Fosforilación , Procesamiento Proteico-Postraduccional , Proteínas Tirosina Fosfatasas/genética , Proteínas Tirosina Fosfatasas/aislamiento & purificación , Proteínas Tirosina Fosfatasas Clase 3 Similares a Receptores , Transducción de SeñalRESUMEN
Toxic epidermal necrolysis (TEN) is a rare life-threatening adverse drug reaction characterized by a massive destruction of the epidermis. Immunohistological studies of skin biopsies of TEN showed infiltrates of predominantly CD8+ T lymphocytes even though other authors reported a prominent involvement of cells of the monocyte-macrophage lineage. The aim of this study was to characterize phenotypically and functionally the cells present in the cutaneous blister fluid of four patients with TEN. We first determined that lymphocytes were predominant in blister fluid obtained early, while monocytes/macrophages later became the most important population. We then showed that this lymphocyte population, mainly CD3+CD8+, corresponded to a peculiar cell subset as they expressed cutaneous leucocyte antigen, killer inhibitory receptors KIR/KAR and failed to express CD28 molecule. Functionally, we determined that blister T lymphocytes had a cytotoxic T lymphocyte (CTL)- and NK-like cytotoxicity. The role of this cytotoxic lymphocyte population present at the site of lesions during TEN remains to be understood.
Asunto(s)
Vesícula/inmunología , Síndrome de Stevens-Johnson/inmunología , Subgrupos de Linfocitos T/inmunología , Adulto , Líquidos Corporales/inmunología , Complejo CD3/metabolismo , Antígenos CD8/metabolismo , Citotoxicidad Inmunológica , Femenino , Humanos , Células Asesinas Naturales/inmunología , Masculino , Persona de Mediana Edad , Receptores Inmunológicos/metabolismo , Receptores KIR , Receptores de Células Asesinas Naturales , Síndrome de Stevens-Johnson/etiología , Linfocitos T Citotóxicos/inmunologíaRESUMEN
DEFINITION: T-cell lymphomas localized predominantly or primarily in skin comprise a heterogeneous group of tumors. The most frequent is fungoid mycosis. T-CELL TUMORS AND GROWTH FACTORS: When growth factors of T-cell tumors, particularly IL-7 and IL-5, were identified, it became possible to grow T-cell tumoral clones in long-term in vitro cultures used to evidence anti-tumoral cellular immune reactions in T-cell skin cancers. Reactional T-cell clones with specific anti-tumoral reactions can be expaned ex vivo. This advance has made it possible to identify tumoral antigens and better understand the mechanisms leading to tumoral escape from anti-tumoral defense systems. THERAPEUTIC PERSPECTIVES: It is now possible to develop immunotherapy strategies based on reinjection of T lymphocytes with specific cytotoxic activity after ex vivo expansion. Injections of immunogenic peptides associated or not with dendritic cells or injection of DNA coding for these tumor antigens are other possibilities.