RESUMEN
CD30 has been proposed to identify Th0/2-type clones. However, the in vivo relevance of this finding is still a matter of debate, as high serum levels of soluble CD30 have been found in both Th1- and Th2- dominated disorders. Among these, rheumatoid arthritis represents a condition where the Th1 predominance is combined with the presence of CD30(+) T-cell activity, particularly in specific stages of the disease. This article discusses the hypothesis that CD30(+) T cells might play a counter-regulatory role at sites of inflammation in Th1-mediated conditions, such as rheumatoid arthritis.
Asunto(s)
Artritis Reumatoide/inmunología , Antígeno Ki-1/biosíntesis , Subgrupos de Linfocitos T/inmunología , Células TH1/inmunología , Animales , Artritis Reumatoide/patología , Humanos , Subgrupos de Linfocitos T/patología , Células TH1/patologíaRESUMEN
INTRODUCTION: Naturally occurring antibodies (auto-Abs) recognizing human granulocyte-colony stimulating factor were detected with high frequency in serum samples obtained from umbilical cord blood of newborns (12 of 65 samples screened) and maternal peripheral blood serum samples from women at the end of gestation (seven of 56 cases tested). The aim of this paper was to demonstrate that auto-Abs anti-G-CSF revealed in the blood of newborns were produced during foetal life. MATERIALS AND METHODS: Mononuclear cells from cord blood samples of different newborns containing high titer anti-G-CSF Abs were infected with Epstein-Barr virus in vitro, and EBV-immortalized B-cell lines were isolated and characterized for specific anti-G-CSF Ab production. RESULTS: Six different, unrelated cell lines of male origin which showed the presence of EBNA-2 antigen in the nucleus, displayed a B-cell phenotype (CD30+, CD5-, CD10-, HLA-DR+, CD19+, CD20+, CD23+, CD38+, CD25+), coexpressed low intensity sIgM and sIgD, and produced only IgM with prevailing lambda clonal restriction and anti-rhG-CSF Ab reactivity. The Ab specificity was proven against either glycosylated or unglycosylated G-CSF by saturable binding in direct enzyme-linked immunosorbent assays, by competition binding and Western immunoblotting assays. CONCLUSION: The secreted Abs did not affect the in vitro generation of granulocyte colonies by human normal adult haemopoietic progenitor cells in soft agar clonogenic assays, suggesting that these Abs were not neutralizing.
Asunto(s)
Autoanticuerpos/biosíntesis , Linfocitos B/inmunología , Sangre Fetal/inmunología , Factor Estimulante de Colonias de Granulocitos/inmunología , Tercer Trimestre del Embarazo/inmunología , Adulto , Autoanticuerpos/inmunología , Linfocitos B/virología , Western Blotting , Línea Celular Transformada , Transformación Celular Viral , Ensayo de Unidades Formadoras de Colonias , Reacciones Cruzadas , Ensayo de Inmunoadsorción Enzimática , Femenino , Sangre Fetal/citología , Glicosilación , Factor Estimulante de Colonias de Granulocitos/química , Granulocitos/citología , Herpesvirus Humano 4/fisiología , Humanos , Inmunidad Innata , Inmunoglobulina D/biosíntesis , Inmunoglobulina D/inmunología , Inmunoglobulina M/biosíntesis , Inmunoglobulina M/inmunología , Inmunofenotipificación , Recién Nacido , Lenograstim , Masculino , Pruebas de Neutralización , Embarazo , Tercer Trimestre del Embarazo/sangre , Proteínas Recombinantes/química , Proteínas Recombinantes/inmunologíaRESUMEN
We detected natural antibodies (auto-Abs) binding human granulocyte-macrophage colony stimulating factor (GM-CSF) in umbilical cord blood (CB) (23 of 94 samples screened) and peripheral blood of women at the end of pregnancy (6 of 42 samples tested). To demonstrate that Abs detected in CB were produced by the fetus, CB mononuclear cells were infected with Epstein-Barr virus in vitro. Ten cell lines producing constitutively anti-recombinant human GM-CSF (rhGM-CSF) Abs were isolated and characterized. These cells displayed a male karyotype, an early activated B cell phenotype, coexpressed surface IgM and IgD, and secreted only IgM with prevailing lambda clonal restriction. Specific cell surface binding of biotinylated rhGM-CSF and high-level anti-rhGM-CSF IgM Ab production were typical features of early cell cultures. In late cell passages the frequency of more undifferentiated B cells increased. Serum Abs of either maternal or fetal origin or Abs produced in culture did not affect the granulocyte and macrophage colony stimulating activity of rhGM-CSF from bone marrow progenitors in soft agar, suggesting that the Abs produced were nonneutralizing.
Asunto(s)
Autoanticuerpos/biosíntesis , Linfocitos B/citología , Linfocitos B/inmunología , Sangre Fetal/citología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/inmunología , Formación de Anticuerpos , Especificidad de Anticuerpos , Línea Celular , Transformación Celular Viral , Técnicas de Cultivo/métodos , Femenino , Células Madre Hematopoyéticas/inmunología , Herpesvirus Humano 4 , Humanos , Inmunoglobulinas/biosíntesis , Recién Nacido , FenotipoAsunto(s)
Antígenos CD/genética , Leucemia Mieloide Aguda/genética , Polimorfismo Genético/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Receptores del Factor de Necrosis Tumoral/genética , Adolescente , Adulto , Anciano , Alelos , Análisis Mutacional de ADN , Femenino , Frecuencia de los Genes , Genotipo , Humanos , Leucemia Mieloide Aguda/inmunología , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Polimorfismo Conformacional Retorcido-Simple , Leucemia-Linfoma Linfoblástico de Células Precursoras/inmunología , Receptores Tipo I de Factores de Necrosis Tumoral , Receptores Tipo II del Factor de Necrosis TumoralRESUMEN
Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a widely expressed EGF superfamily member that induces mitogenic and/or chemotactic activities toward different cell types through binding to EGF receptors 1 or 4. Membrane-bound HB-EGF exerts growth activity and adhesion capabilities and possesses the unique property of being the receptor for diphtheria toxin (DT). Using molecular and functional techniques, we show that human polymorphonuclear granulocytes (PMN), which did not express HB-EGF in resting conditions, expressed it at mRNA and protein level, following incubation with granulocyte-macrophage colony-stimulating factor (GM-CSF). Other classic agonists for PMN (including lipopolysaccharide, phagocytable particles, tumor necrosis factor-alpha, or G-CSF) failed to induce HB-EGF. The effects of GM-CSF on HB-EGF mRNA levels were concentration-dependent, reached a plateau after 1 to 2 hours of stimulation, and did not require protein synthesis. After GM-CSF treatment, membrane-bound HB-EGF was detected by flow cytometry. At the same time, PMN acquired sensitivity to the apoptosis-promoting effect of DT, which, moreover, specifically suppressed the GM-CSF-induced priming of formyl-methionyl-leucyl-phenylalanine-stimulated superoxide anion release. Finally, soluble HB-EGF was detected in the PMN culture medium by a specific enzyme-linked immunosorbent assay. Thus, we provide evidence that HB-EGF is specifically inducible by GM-CSF in PMN and represents a novel peptide to be included in the repertoire of PMN-derived cytokines.
Asunto(s)
Toxina Diftérica/farmacología , Factor de Crecimiento Epidérmico/biosíntesis , Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Neutrófilos/metabolismo , Receptores de Superficie Celular/biosíntesis , Citometría de Flujo , Factor de Crecimiento Similar a EGF de Unión a Heparina , Humanos , Péptidos y Proteínas de Señalización Intercelular , ARN Mensajero/biosíntesisRESUMEN
BACKGROUND: Patients completely asymptomatic with extremely high levels of IgE have rarely been reported. One such case, in which the immunophenotype pattern of lymphocyte subsets and their cytokine profile were investigated, is described here. OBJECTIVE: To assess whether the cytokine production was consistent with a T helper 2-type immune response, as suggested by theories regarding the functional polarization of helper and cytotoxic T cells in hyper-IgE conditions. METHODS: An asymptomatic 79-year-old man presented with persistent high levels of serum IgE and sporadic hypereosinophilia without any evidence of an underlying pathologic condition. We investigated the immunophenotype of circulating lymphocytes, the expression/release of CD30 (a member of the tumor necrosis factor receptor family preferentially associated with T helper 2-type immune responses) and the intracellular patterns of interferon-gamma (IFN-gamma), Interleukin-2 (IL-2), IL-4, IL-5, and IL-10 production by T cell subsets, as evaluated by single-cell flow-cytometric analysis. RESULTS: The majority of lymphocytes displayed the membrane immunophenotype of NK cells. Both CD4+ and CD8+ T cells were reduced and expressed the "memory" (CD4+/CD45RO+) and the "naive" (CD8+/CD45RA+) phenotypes, respectively. Among CD4+ T cells, CD30 expression was increased in the resting condition and was further inducible following stimulation with mitogenic anti-CD3. Interleukin-4, IL-2, and IL-10 production by CD4+ T cells was increased, whereas IFN-gamma was reduced as compared with normals. CONCLUSIONS: The data suggest that a polarization of CD4+ T cells towards a T helper 0/2-type cytokine pattern occurred in this patient in spite of CD4+ cell reduction and NK cell expansion.
Asunto(s)
Antígenos de Superficie/genética , Citocinas/biosíntesis , Inmunoglobulina E/sangre , Líquido Intracelular/química , Síndrome de Job/sangre , Linfocitos/inmunología , Anciano , Linfocitos T CD4-Positivos/citología , Linfocitos T CD4-Positivos/inmunología , Humanos , Inmunofenotipificación , Interferón gamma/sangre , Interleucina-4/sangre , Antígeno Ki-1/sangre , Células Asesinas Naturales/citología , Recuento de Linfocitos , MasculinoRESUMEN
In order to investigate the T cell cytokine profile during age-dependent maturation of the immune response, we evaluated the cytokine expression of CD4+ and CD8+ circulating cells by flow cytometric single-cell analysis after non-specific stimulation in vitro in different age groups of normal individuals, from cord blood to adulthood. Moreover, we correlated these lymphocyte cytokine patterns with the expression/release of CD30, a member of the tumour necrosis factor (TNF) receptor superfamily, which has been suggested to be related to the T helper/cytotoxic (Th(c))2-type immune responses, in order to verify this association in vivo, in non-pathological conditions. The results showed a progressive increase of circulating Th(c)1-type, interferon-gamma (IFN-gamma)- and/or IL-2-producing T cells along with ageing and, conversely, a stable number, although higher than in cord blood samples, of CD4+/IL-4+ T cells in the post-natal groups. In addition, serum levels of soluble CD30 (sCD30) and numbers of circulating CD4+/CD30+ and CD8+/CD30+ T cells were significantly higher in children aged < 5 years in comparison with those found either in cord blood or in blood from both older children and adults. These data support the concept of a progressive polarization of the Th(c) cell cytokine profile towards the Th(c)1 pattern during age-dependent maturation of the immune response. Moreover, the peak of CD30 expression/release in early infancy before the Th(c)1 shifting occurs, although not associated with a significant increase of circulating IL-4+ T cells, raises the question of the possible relationship in vivo between CD30 and Th(c)2-type immune responses.
Asunto(s)
Envejecimiento/inmunología , Citocinas/biosíntesis , Citotoxicidad Inmunológica , Antígeno Ki-1/biosíntesis , Antígeno Ki-1/sangre , Subgrupos de Linfocitos T/metabolismo , Células TH1/metabolismo , Adolescente , Adulto , Diferenciación Celular/inmunología , Membrana Celular/inmunología , Membrana Celular/metabolismo , Células Cultivadas , Niño , Preescolar , Sangre Fetal , Humanos , Lactante , Líquido Intracelular/inmunología , Líquido Intracelular/metabolismo , Solubilidad , Subgrupos de Linfocitos T/inmunología , Células TH1/inmunologíaRESUMEN
BACKGROUND AND OBJECTIVE: Measles virus infection (MVI) has been reported to be characterized by an imbalanced Th(1/2)-type cytokine profile. CD30 has been proposed as a receptor preferentially associated with the Th(0/2)-type cytokine pattern. The aim of this study was therefore to define the peripheral T lymphocyte cytokine profile and to test which CD30 expression pattern it was associated with in MVI. DESIGN AND METHODS: The design of the study was a prospective evaluation with comparative analysis. The serum levels of the soluble form of CD30 (sCD30) were determined at diagnosis and at weekly intervals up to 4 weeks, using an ELISA, in 23 males (median age 19), who developed MVI while serving in the Italian army and who were admitted to the Infectious Disease Unit of the Military Hospital in Padua. In 10 of the patients at diagnosis we studied the lymphoid immunophenotype and, after non-specific ex vivo stimulation, the expression of IFNgamma, IL-2 and IL-4 by peripheral T cells using flow cytometry single cell analysis. In 3 patients such evaluations were also performed 7 weeks later. RESULTS: At diagnosis, we found (i) reduction of IFNgamma+/CD4+ T cells (p=0.048 vs controls) in the absence of substantial variation of IL-2+ and IL-4+ T cells (p=ns vs controls); (ii) expansion of CD30+/ CD4+ and CD30+/CD8+ T cell subsets (p<0.01 vs controls); (iii) high sCD30 values (median 61 U/mL; p<0.001 vs controls); (iiii) a context of lymphopenia (0. 728+/-0.292 lymph x10(9)/L). sCD30 remained elevated up to 4 weeks from MVI onset [median values 53, 49, 50, 34 U/mL after 1, 2, 3 and 4 weeks, respectively (p=ns between different time points)]. In 3 patients tested 7 weeks after diagnosis, we still observed decreased IFNgamma production by CD4+ and CD8+ T cells (p=0.05 and <0.01, respectively vs controls) and reduction of CD4+ and CD8+/IL-2+ T cells (p<0.01). INTERPRETATION AND CONCLUSIONS: MVI was characterized by featuresof inadequate Th/Tc(1) activation associated with increased circulating CD30+ T cells and elevated sCD30 levels, supporting a correlation between Th/Tc status and CD30 expression/release pattern in vivo.
Asunto(s)
Interferón gamma/inmunología , Interleucina-2/inmunología , Interleucina-4/inmunología , Antígeno Ki-1/inmunología , Sarampión/inmunología , Subgrupos de Linfocitos T/inmunología , Adulto , Citometría de Flujo , Humanos , Inmunofenotipificación , Interferón gamma/biosíntesis , Interleucina-2/biosíntesis , Interleucina-4/biosíntesis , Antígeno Ki-1/biosíntesis , MasculinoRESUMEN
Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is an EGF family member expressed by numerous cell types that binds to EGF receptor 1 (HER-1) or 4 (HER-4) inducing mitogenic and/or chemotactic activities. Membrane-bound HB-EGF retains growth activity and adhesion capabilities and the unique property of being the receptor for diphtheria toxin (DT). The interest in studying HB-EGF in acute leukemia stems from these mitogenic, chemotactic, and receptor functions. We analyzed the expression of HB-EGF in L428, Raji, Jurkat, Karpas 299, L540, 2C8, HL-60, U937, THP-1, ML-3, and K562 cell lines and in primary blasts from 12 acute myeloid leukemia (AML) cases, by reverse-transcriptase polymerase chain reaction (RT-PCR) and Northern blot and by the evaluation of sensitivity to DT. The release of functional HB-EGF was assessed by evaluation of its proliferative effects on the HB-EGF-sensitive Balb/c 3T3 cell line. HB-EGF was expressed by all myeloid and T, but not B (L428, Raji), lymphoid cell lines tested, as well as by the majority (8 of 12) of ex vivo AML blasts. Cell lines (except for the K562 cell line) and AML blasts expressing HB-EGF mRNA underwent apoptotic death following exposure to DT, thus demonstrating the presence of the HB-EGF molecule on their membrane. Leukemic cells also released a fully functional HB-EGF molecule that was mitogenic for the Balb/c 3T3 cell line. Factors relevant to the biology of leukemic growth, such as tumor necrosis factor-alpha (TNF-alpha), 1alpha,25-(OH)2D3, and especially all-trans retinoic acid (ATRA), upregulated HB-EGF mRNA in HL-60 or ML-3 cells. Granulocyte-macrophage colony-stimulating factor (GM-CSF) induced HB-EGF mRNA and acquisition of sensitivity to DT in one previously HB-EGF-negative leukemia case. Moreover, the U937 and Karpas 299 cell lines expressed HER-4 mRNA. This work shows that HB-EGF is a growth factor produced by primary leukemic cells and regulated by ATRA, 1alpha, 25-(OH)2D3, and GM-CSF.
Asunto(s)
Factor de Crecimiento Epidérmico/biosíntesis , Leucemia Mieloide/metabolismo , Enfermedad Aguda , Adulto , Anciano , Apoptosis , Femenino , Células HL-60 , Factor de Crecimiento Similar a EGF de Unión a Heparina , Humanos , Péptidos y Proteínas de Señalización Intercelular , Células Jurkat , Células K562 , Leucemia Mieloide/patología , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , ARN Mensajero/análisis , Células U937RESUMEN
The tumour necrosis factor (TNF)/TNF-receptor (TNFR) complex plays a role in the growth of leukaemic cells. We retrospectively investigated the relationship between pretreatment serum concentration of soluble TNFR (p55- and p75-sTNFRs) and outcome in adult acute myeloid (AML 82 cases) and lymphoid (ALL 44 cases) leukaemia. Both sTNFRs were significantly higher in AML (p55-sTNFR 4.53 +/- 3.7, median 3.75; p75-sTNFR 6.51 +/- 5.25 ng/ml, median 4.72) and ALL sera (3.31 +/- 1.5, median 2.95; 5.30 +/- 2.3 ng/ml, median 4.56, respectively) than in controls (1.89 +/- 0.5, median 1.98; 2.22 +/- 0.8 ng/ml, median 2.37) (P < 0.01 for both sTNFRs). Fresh leukaemic cells expressed p55- and p75-sTNFRs, which were modulated and released into the supernatant (SN) following short-term in vitro culture, suggesting that in vivo sTNFRs were also leukaemia-derived. Whereas no correlation was observed between sTNFRs and outcome in ALL, in AML higher p55-sTNFR levels (> 3.75 ng/ml) were associated with shorter disease-free survival (DFS) (P = 0.006) and overall survival (OS) (P = 0.0004). At multivariate analysis p55-sTNFR was the most significant predictor of DFS (P = 0.006) and OS (P < 0.001). Our data suggest that the prognostic significance of p55-sTNFR in AML could be related to relevant biological features of AML blasts.
Asunto(s)
Antígenos CD/sangre , Biomarcadores de Tumor/sangre , Leucemia Mieloide/sangre , Leucemia-Linfoma Linfoblástico de Células Precursoras/sangre , Receptores del Factor de Necrosis Tumoral/sangre , Adolescente , Adulto , Anciano , Supervivencia sin Enfermedad , Femenino , Estudios de Seguimiento , Humanos , Leucemia Mieloide/terapia , Masculino , Persona de Mediana Edad , Análisis Multivariante , Leucemia-Linfoma Linfoblástico de Células Precursoras/terapia , Pronóstico , Receptores Tipo I de Factores de Necrosis Tumoral , Receptores Tipo II del Factor de Necrosis Tumoral , Estudios Retrospectivos , Solubilidad , Tasa de Supervivencia , Resultado del TratamientoRESUMEN
Cyclin-dependent kinase-6 (CDK6) is the earliest inducible member of the CDK family in human T lymphocytes, involved in growth factor stimulation and cell cycle progression. CDK6 is one of the targets of p16 and p15, CDK inhibitors encoded by MTS1 and MTS2, two tumor suppressor genes that are frequently deleted in T-cell leukemia. In this study we have investigated CDK6 expression in normal and neoplastic lymphoid tissues using immunohistochemistry and flow cytometry. In normal (six samples) and hyperplastic (four samples) thymuses, strong CDK6 expression was observed in a discrete proportion of cortical thymocytes (10 to 15%), mainly located in the peripheral (subcapsular) zone of the cortex. All tested cases of T-cell lymphoblastic lymphoma/leukemia (T-LBL/ALL) showed strong CDK6 expression in the majority (up to 100%) of neoplastic lymphoid cells. Western blot analysis confirmed the expected CDK6 protein size (40 kd). According to Southern blot analysis, CDK6 overexpression in neoplastic T lymphoblasts was not due to gene amplification. In all other lymphomas investigated (28 peripheral T-cell non-Hodgkin's lympohomas (T-NHLs), 7 CD30+ anaplastic NHLs, 22 high-grade B-NHLs, 15 low-grade B-NHLs, 25 B-cell precursor ALLs), CDK6 was not expressed or expressed at low levels, with the only exception of three nasal angiocentric T-NHLs, all exhibiting CDK6 immunoreactivity comparable to that observed in T-LBL/ALL. These data provide evidence that CDK6 is abnormally expressed in T-LBL/ALL and may be involved in the pathogenesis of this malignancy. In addition, the quantitative difference of CDK6 expression between neoplastic and non-neoplastic cortical thymocytes can be potentially useful in the differential diagnosis of thymic neoplasms on histological and cytological specimens.
Asunto(s)
Quinasas Ciclina-Dependientes , Leucemia/enzimología , Linfoma de Células T/enzimología , Leucemia-Linfoma Linfoblástico de Células Precursoras/enzimología , Proteínas Serina-Treonina Quinasas/metabolismo , Timo/metabolismo , Southern Blotting , Western Blotting , Quinasa 6 Dependiente de la Ciclina , Feto/citología , Feto/metabolismo , Citometría de Flujo , Humanos , Lactante , Ganglios Linfáticos/enzimología , Linfoma de Células B/enzimología , Valores de Referencia , Timoma/enzimología , Timo/citología , Timo/embriología , Neoplasias del Timo/enzimologíaRESUMEN
CD30 has been suggested to play a role in HIV infection. In this study the serum concentration of soluble CD30 (sCD30) was determined by an ELISA essay on samples collected from patients with acute primary HIV-1 infection during the acute phase (n = 17) and after seroconversion (n = 13). sCD30 during acute infection was consistently elevated (137.58 +/- 120.33 versus 6.4 +/- 5.4 U/ml (mean +/- s.d.) in normal controls, P<0.0001) and decreased after seroconversion (49.1 +/- 66.17 U/ml; P = 0.0018 compared with acute infection). This trend mirrored the disappearance of detectable levels of HIV antigen in the blood, resulting in a direct correlation between sCD30 and HIVAg values (P = 0.002). These data suggest that the high levels of sCD30 observed during the peak concentration of HIVAg in acute primary HIV infection might reflect the high rate of viral replication.
Asunto(s)
Infecciones por VIH/inmunología , VIH-1 , Antígeno Ki-1/sangre , Enfermedad Aguda , Adulto , Femenino , Antígenos VIH/sangre , Infecciones por VIH/sangre , Infecciones por VIH/virología , VIH-1/inmunología , Humanos , Masculino , Persona de Mediana Edad , SolubilidadRESUMEN
Hodgkin's disease (HD) is characterised by a complex architectural and functional derangement of involved tissues. The interactions between neoplastic cells and the heterogeneous microenvironment lead to the expression and release of different cellular messengers [cytokines, soluble (s) forms of cytokine receptors and other membrane-associated molecules] which can be detected in the circulation and evaluated as biological markers. We and others investigated several of these molecules looking for their possible role as diagnostic or prognostic parameters in patients with HD. We update here the results of serum determination of sIL-2Ralpha, sCD8, sICAM-1, sTNFRs, and sCD30 in a large series of cases from our institution. We found that their levels are generally increased at presentation and during the active phase of the disease. They correlate with stage and clinical aggressiveness and have some prognostic implication. However, we were unable to demonstrate a prognostic usefulness for their detection, with the exception for sCD30 which was found to directly correlate with disease spread and burden at presentation and, most importantly, to have an independent prognostic significance. The prognostic significance of sCD30 might derive from a crucial involvement of this molecule in the pathophysiology of HD.
Asunto(s)
Biomarcadores de Tumor/sangre , Enfermedad de Hodgkin/sangre , Adulto , Antígenos CD8/sangre , Enfermedad de Hodgkin/diagnóstico , Enfermedad de Hodgkin/metabolismo , Humanos , Molécula 1 de Adhesión Intercelular/sangre , Antígeno Ki-1/sangre , Pronóstico , Receptores de Interleucina-2/sangre , Receptores del Factor de Necrosis Tumoral/sangreRESUMEN
BACKGROUND: In myeloid blasts, the expression and release of the multifunctional chemokine IL-8 could be expected to be differentiation-associated. METHODS: We investigated the profile of interleukin-8 (IL-8) expression and release by leukemic cells obtained at diagnosis from 42 untreated adult patients with acute myeloid leukemia of various FAB subtypes (2 M0, 7 M1, 6 M2, 6 M3, 10 M4 and 11 M5). IL-8 transcripts were evaluated by Northern blot and densitometric analysis. IL-8 release by myeloid blasts was evaluated by a specific ELISA either in sera at diagnosis or in supernatants (SN) obtained from cultured leukemic cells. RESULTS: In basal conditions, Northern blot analysis revealed detectable IL-8 transcripts in 15/29 cases, eleven of which were classified as M4-M5 and 4 as FAB M0-M3. Densitometric analysis of IL-8 transcript bands showed higher expression in M4-M5 than in M0-M3 cases (mean values +/- SD: 16.5 +/- 21 and 0.77 +/- 1.36 densitometric units, respectively; p = 0.012). Higher IL-8 serum levels were observed in leukemic patients as opposed to normal controls (mean values +/- SD: 0.53 +/- 0.75 vs 0.003 +/- 0.014 ng/mL, respectively; p = 0.006). Furthermore, a trend (though not of statistical significance) towards higher IL-8 serum values was observed in M4-M5 as opposed to M0-M3 subtypes. After 24 hours of culture, the majority of myeloid blasts (95%) spontaneously released detectable amounts of IL-8 into SN. However, M4-M5 released substantially higher amounts of IL-8 than M0-M3 blasts (mean +/- SD: 68 +/- 46 and 8.5 +/- 12 ng/mL, respectively; p < 0.001). This difference between M0-M3 and M4-M5 blasts was already observed after 6 hours of culture and increased over 72 hours. CONCLUSIONS: Our findings confirm and further support the preferential release of high levels of IL-8 by myeloid blasts showing monocytic differentiation.
Asunto(s)
Interleucina-8/metabolismo , Leucemia Mieloide/metabolismo , Monocitos/citología , Enfermedad Aguda , Adolescente , Adulto , Anciano , Diferenciación Celular/fisiología , Humanos , Leucemia Mieloide/patología , Persona de Mediana Edad , Células Tumorales CultivadasRESUMEN
There is evidence that both cellular and humoral components of the immune response are required for viral clearance to occur in chronic hepatitis B. Recent studies demonstrated that CD30 molecule, a member of the tumour necrosis factor superfamily of membrane cytokine receptors, is expressed on, and released as a soluble molecule (sCD30) by activated T cells producing T helper 2 (Th2) cytokines, which modulate antibody responses. To better characterize the immunoregulatory mechanisms in chronic hepatitis B virus (HBV) infection, sCD30 values were evaluated by an ELISA in 90 hepatitis B surface (HBsAg)-positive patients with chronic hepatitis, selected on the basis of active viral replication and biochemical activity. At presentation abnormal levels (> 20 U/ml) of sCD30 were detected in 57 (63%) out of 90 patients with chronic hepatitis B, and median value was significantly higher in this group of patients compared with that of healthy HBsAg carriers (26.7 versus 10.5 U/ml, P < 0.000 05) and with normal controls (26.7 versus 3 U/ml P < 0.000 01). Sequential studies of chronic hepatitis B did confirm the association of raised sCD30 levels with the active phase of the illness. On the other hand, a significant decrease was noted when sCD30 levels at diagnosis and after termination of HBV replication and biochemical remission of hepatitis were compared in 10 untreated patients (median, 28 U/ml at entry versus 8 U/ml at remission, P < 0.01) and in six patients responding to interferon-alpha therapy (median, 29.5 U/ml at entry versus 6 U/ml at remission, P < 0.05). The high serum sCD30 levels reported during the active phase of HBsAg-positive chronic hepatitis suggest a certain degree of immune competence of these patients, at least with respect to a Th2-type response. These data are in agreement with recent serologic surveys showing that most chronic hepatitis B patients do demonstrate ongoing humoral immune response to HBV antigens, using novel immunoassays designed to detect antibody in the presence of excess serum viral antigen. Th2 functions that mainly promote humoral immunity to HBV antigens may be critical, in association with a competent virus-specific cytotoxicity, for efficient termination of HBV replication in chronic hepatitis B.
Asunto(s)
Hepatitis B/inmunología , Antígeno Ki-1/sangre , Adolescente , Adulto , Anciano , Femenino , Hepatitis B/sangre , Hepatitis B/patología , Hepatitis Crónica/sangre , Hepatitis Crónica/inmunología , Hepatitis Crónica/patología , Humanos , Masculino , Persona de Mediana Edad , SolubilidadRESUMEN
The two tumor necrosis factor receptors (TNF-Rs) exist in their soluble form in different biological fluids. In this study we investigated the concentrations of soluble tumor necrosis factor receptors (sTNF-Rs) in the culture supernatants of leukemic cells and in the serum obtained from 33 patients: 12 with hairy cell leukemia (HCL) and 21 with B cell chronic lymphocytic leukemia (B-CLL). In seven patients with HCL, sTNF-Rs were also evaluated following in vivo treatment with interferon-alpha (IFN-alpha). Purified leukemic cells from patients with HCL and B-CLL spontaneously released sTNF-R75 but not sTNF-R55. The levels of sTNF-R75 were higher in supernatants obtained from cultured hairy cells than from cultures of B-CLL cells. The shedding of sTNF-R75 was further increased both in HCL and B-CLL subjects by some B cell-related stimuli, including BCGF, PMA, SAC and was partially inhibited by IFN-alpha in patients with HCL. Sera from HCL patients presented increased levels of both sTNF-Rs with respect to normal controls. Treatment of HCL patients with IFN-alpha resulted in a decrease in serum levels of sTNF-Rs, particularly sTNF-R75. These findings suggest that leukemic cells account for the increased serum levels of sTNF-R75 observed in patients with HCL and B-CLL, but not for sTNF-R55.
Asunto(s)
Interferón-alfa/uso terapéutico , Leucemia de Células Pilosas/inmunología , Leucemia Linfocítica Crónica de Células B/inmunología , Receptores del Factor de Necrosis Tumoral/biosíntesis , Antígenos CD/análisis , Humanos , Leucemia de Células Pilosas/sangre , Leucemia de Células Pilosas/terapia , Leucemia Linfocítica Crónica de Células B/sangre , Leucemia Linfocítica Crónica de Células B/terapia , Células Tumorales CultivadasRESUMEN
PURPOSE: To determine serum levels of the soluble form of CD30 molecule (sCD30) in patients with Ki-1/CD30+ anaplastic large-cell lymphoma (ALCL), and to evaluate its correlation with clinical features at presentation and its possible role as a tumor marker to monitor response to treatment and subsequent follow-up. PATIENTS AND METHODS: sCD30 serum levels were measured with an improved commercial sandwich enzyme-linked immunosorbent assay (ELISA) test kit in 24 patients with CD30+ ALCL at diagnosis and in 13 after treatment. RESULTS: Increased values (> 20 U/mL) at diagnosis were observed in 23 of 24 cases (median, 842.5 U/mL; range, 16 to 37,250) as compared with controls (P < .0001). These values were greater than those of 60 stage-matched cases of Hodgkin's disease (HD) (P < .0001). The highest median value was observed in patients with T-cell-type ALCL (1,690 U/mL), with a significant overall difference as compared with B- and null-cell types (P = .004). Phenotype maintained its significance when results were corrected for other parameters, such as age, sex, and stage (P = .03). sCD30 values returned to the normal range in complete remission (CR), but remained increased in one patient who only partially responded to treatment. Subsequent increases of sCD30 levels were recorded in four of four patients after relapse. CONCLUSION: sCD30 appears to be a new biologic serum tumor marker of possible use in the clinical setting of CD30+ ALCL.
Asunto(s)
Biomarcadores de Tumor/sangre , Antígeno Ki-1/sangre , Linfoma Anaplásico de Células Grandes/sangre , Adolescente , Adulto , Anciano , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Niño , Femenino , Enfermedad de Hodgkin/sangre , Humanos , Linfoma Anaplásico de Células Grandes/tratamiento farmacológico , Linfoma Anaplásico de Células Grandes/patología , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Inducción de RemisiónRESUMEN
OBJECTIVE: To measure the levels of serum soluble CD30 (sCD30), a marker of cells producing T helper 2(Th2)-type cytokines, in systemic lupus erythematosus (SLE) and undifferentiated connective tissue disease (UCTD), and to determine its value in assessing disease activity. METHODS: Serum levels of sCD30 were measured by ELISA in 21 patients with SLE, in 17 patients with UCTD and in 40 normal donors. Disease activity was evaluated according to the ECLAM scoring system. RESULTS: sCD30 values were 53.84 +/- 58.24 U/mL in SLE, 22.65 +/- 9.82 U/mL in UCTD and 5.3 +/- 5.7 in normal controls (p < 0.0005 SLE vs controls; p < 0.05 SLE vs UCTD). sCD30 levels were directly related to the disease activity (p < 0.002). CONCLUSION: These data support a relationship between the Th2-type immune response and the pathogenesis of SLE, and suggest that sCD30 can be used as a simple marker for the evaluation of disease activity.
Asunto(s)
Citocinas/biosíntesis , Antígeno Ki-1/sangre , Lupus Eritematoso Sistémico/inmunología , Linfocitos T Colaboradores-Inductores/metabolismo , Adolescente , Adulto , Anticuerpos Antinucleares/sangre , Progresión de la Enfermedad , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Masculino , Persona de Mediana Edad , Linfocitos T Colaboradores-Inductores/inmunologíaRESUMEN
One hundred and twenty-four sera from patients with various leukemic B-cell chronic lymphoproliferative diseases were investigated at diagnosis by ELISA for their soluble CD23 content. Immunophenotyping was carried out in all patients, and in a selected subset the mean number of membrane-bound CD23 molecules per cell was also investigated. Seventy-three patients had typical B chronic lymphocytic leukemia, 41 leukemic B-cell disorders with atypical morphological and/or immunophenotypic features, 5 had low-grade follicular cell lymphoma in the leukemic phase, and 5 had splenic lymphoma with villous lymphocytes. Soluble CD23 levels were significantly higher than in normal sera (mean +/- SD: typical B chronic lymphocytic leukemia 3,650 +/- 4,654 U/ml, atypical B chronic lymphocytic leukemia 3,440 +/- 4,671 U/ml, follicular cell lymphoma 3,200 +/- 1,511 U/ml, splenic lymphoma with villous lymphocytes 8,236 +/- 7,294 U/ml, controls 137 +/- 128 U/ml; P < 0.001). More advanced Rai's stages were related to higher soluble CD23 levels (P < 0.01), both in typical and atypical B chronic lymphocytic leukemias, the highest levels and the best correlation with the absolute number of circulating CD19+ cells (r = 0.50) being observed in the typical form. The number of membrane-bound CD23 molecules per cell was significantly higher in typical than in atypical B chronic lymphocytic leukemias (mean number 156,727 +/- 94,668 vs. 12,010 +/- 10,643, P < 0.001). Our data suggest that soluble CD23 levels correlate with the clinical and biological features of leukemic B-cell lymphoproliferative disorders.