RESUMEN
Human leukocyte antigen-G (HLA-G) molecules are non-classical HLA class I antigens with an important role in pregnancy immune regulation and inflammation control. Soluble HLA-G proteins can be generated through two mechanisms: alternative splicing and proteolytic release, which is known to be metalloprotease mediated. Among this class of enzymes, matrix metalloproteinases (MMPs) might be involved in the HLA-G1 membrane cleavage. Of particular interest are MMP-2 and MMP-9, which regulate the inflammatory process by cytokine and chemokine modulation. We evaluated the effect of MMP-9 and MMP-2 on HLA-G1 membrane shedding. In particular, we analyzed the in vitro effect of these two gelatinases on the secretion of HLA-G1 via proteolytic cleavage in 221-G1-transfected cell line, in JEG3 cell line, and in peripheral blood mononuclear cells. The results obtained by both cell lines showed the role of MMP-2 in HLA-G1 shedding. On the contrary, MMP-9 was not involved in this process. In addition, we identified three possible highly specific cleavage sites for MMP-2, whereas none were detected for MMP-9. This study suggests an effective link between MMP-2 and HLA-G1 shedding, increasing our knowledge on the regulatory machinery beyond HLA-G regulation in physiological and pathological conditions.
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Membrana Celular/metabolismo , Antígenos HLA-G/metabolismo , Metaloproteinasa 2 de la Matriz/metabolismo , Proteolisis , Secuencia de Aminoácidos , Anticuerpos Bloqueadores/farmacología , Línea Celular Tumoral , Membrana Celular/efectos de los fármacos , Cicloheximida/farmacología , Ácido Edético/farmacología , Antígenos HLA-G/química , Humanos , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/metabolismo , Metaloproteinasa 2 de la Matriz/química , Metaloproteinasa 9 de la Matriz/química , Metaloproteinasa 9 de la Matriz/metabolismo , Datos de Secuencia Molecular , Proteolisis/efectos de los fármacosRESUMEN
BACKGROUND AIMS: The beneficial activity of mesenchymal stromal cells (MSC) in allogeneic hematopietic stem cell transplantation requires correct use in terms of cell dose and timing of infusion and the identification of biomarkers for selection. The immunosuppressive bone marrow (BM)-derived MSC (BM-MSC) functions have been associated with the production of soluble HLA-G molecules (sHLA-G) via interleukin (IL)-10. We have established a reliable method for evaluating BM-MSC HLA-G expression without the influence of peripheral blood mononuclear cells (PBMC). METHODS: Thirteen BM-MSC from donors were activated with recombinant IL-10 or co-cultured with 10 different phytohemagglutinin (PHA)-treated PBMC (PHA-PBMC). Membrane-bound and sHLA-G expression was evaluated by flow cytometry and enzyme-linked immunosorbent assay (ELISA), respectively; lymphoproliferation was measured by (methyl-(3)H)thymidine. RESULTS: The results demonstrated the ability of IL-10 to stimulate both membrane-bound and sHLA-G production by BM-MSC. The levels of HLA-G expression induced by IL-10 in BM-MSC were associated with the inhibition of PHA-PBMC proliferation (sHLA-G, P = 0.0008, r = 0.9308; membrane HLA-G, P = 0.0005, r = 0.9502). CONCLUSIONS: We propose the evaluation of sHLA-G production in IL-10-treated BM-MSC cultures as a possible marker of immunoregulatory function.
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Células de la Médula Ósea/inmunología , Separación Celular/métodos , Antígenos HLA/análisis , Antígenos de Histocompatibilidad Clase I/análisis , Tolerancia Inmunológica , Terapia de Inmunosupresión , Células Madre Mesenquimatosas/inmunología , Adulto , Células de la Médula Ósea/efectos de los fármacos , Células Cultivadas , Técnicas de Cocultivo , Antígenos HLA/biosíntesis , Antígenos HLA-G , Antígenos de Histocompatibilidad Clase I/biosíntesis , Humanos , Interleucina-10/farmacología , Leucocitos Mononucleares/inmunología , Activación de Linfocitos , Células Madre Mesenquimatosas/efectos de los fármacos , Persona de Mediana Edad , Fitohemaglutininas/farmacología , Células del Estroma/efectos de los fármacos , Células del Estroma/inmunologíaRESUMEN
Bacterial LPS induces the release of ATP from immune cells. Accruing evidence suggests that extracellular ATP participates in the inflammatory response as a proinflammatory mediator by activating the inflammasome complex, inducing secretion of cytokines (IL-1, IL-18) and cell damaging agents such as oxygen radicals, cationic proteins, and metalloproteases. It is not known whether ATP can also act as a proinflammatory mediator by inhibiting production of molecules down-modulating the immune response. Here, we show that extracellular ATP impairs in an IL-10-dependent fashion the expression of the tolerogenic soluble and membrane-bound HLA-G Ag in human monocytes. The effect of ATP was mimicked by BzATP (3'-O-(4-benzoyl)benzoyl-ATP) and greatly reduced by pretreatment with oATP (periodate-oxidized ATP), KN-62 (1-[N,O-bis(5-isoquinoline-sulfonyl)-N-methyl-l-tyrosyl]-4-phenylpiperazine), and an anti-P2X(7) mAb, thus pointing to a specific role of the P2X(7) receptor. The effect of ATP was time- and dose-dependent and was not due to a decrease in expression of IL-10 receptor. Inhibition by ATP was reverted by supplementation of culture medium with exogenous IL-10. Due to the well-known immunosuppressive activity of IL-10 and soluble HLA-G, this novel effect of ATP might be relevant for the pathophysiology and therapy of inflammatory disorders.
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Regulación hacia Abajo/inmunología , Espacio Extracelular/inmunología , Antígenos HLA/inmunología , Antígenos HLA/metabolismo , Antígenos de Histocompatibilidad Clase I/inmunología , Antígenos de Histocompatibilidad Clase I/metabolismo , Mediadores de Inflamación/fisiología , Monocitos/inmunología , Monocitos/metabolismo , Receptores Purinérgicos P2/metabolismo , Línea Celular Tumoral , Membrana Celular/inmunología , Membrana Celular/metabolismo , Células Cultivadas , Espacio Extracelular/metabolismo , Antígenos HLA/biosíntesis , Antígenos HLA-G , Antígenos de Histocompatibilidad Clase I/biosíntesis , Humanos , Mediadores de Inflamación/metabolismo , Interleucina-10/antagonistas & inhibidores , Interleucina-10/biosíntesis , Interleucina-10/metabolismo , Lipopolisacáridos/sangre , Receptores Purinérgicos P2/fisiología , Receptores Purinérgicos P2X7RESUMEN
HLA-G antigens are non-classical HLA-class I anti-inflammatory molecules. Since styrene exposure has been suggested to induce immune alteration, we analyzed plasma levels and "in vitro" peripheral blood mononuclear cell (PBMC) production of soluble HLA-G (sHLA-G) and interleukin-10 (IL-10) molecules after lipopolysaccharide (LPS) stimulation, in styrene exposed workers and healthy subjects. Exposed workers showed reduced plasma levels of sHLA-G and IL-10 in comparison to healthy controls. Similarly, lower levels of sHLA-G and IL-10 molecules were observed in PBMC culture supernatants after LPS activation. These data propose styrene exposure as a mediator of impaired sHLA-G production.
RESUMEN
BACKGROUND AND STUDY AIMS: Functional dyspepsia is an exclusion diagnosis requiring different tests, including endoscopy, often repeated over time. Duodenal biopsies are frequently resorted to, not rarely revealing duodenal microscopic inflammation. Aim of the study is to confirm a previously supposed role of antro-duodenal low-grade inflammation in functional dyspepsia, evaluating the frequency of duodenal lymphocytosis, H. pylori infection and their association in a group of patients with functional dyspepsia compared to asymptomatic control subjects. PATIENTS AND METHODS: A cross-sectional, observational study has been conducted screening all the patients who underwent duodenal biopsies during upper endoscopy, in a 30â¯months period. All the patients without endoscopic lesions were analysed. The study group consisted of patients compatible with the diagnosis of functional dyspepsia (Rome III criteria). The control group consisted of healthy asymptomatic subjects in the population subjected to endoscopy. The presence of duodenal lymphocytosis and of H. pylori infection in the two groups was evaluated. RESULTS: 216 patients were enrolled: 161 in the functional dyspepsia group and 55 as asymptomatic control group. The frequency of duodenal lymphocytosis was similar between cases and control groups (25.47% vs 25.45%; pâ¯=â¯0.99), as well as H. pylori infection (26.71% vs 23.64%; pâ¯=â¯0.78). Duodenal lymphocytosis was significantly associated with functional dyspepsia only in H. pylori positive dyspeptic patients (pâ¯=â¯0.047). 94% of the subjects with both lymphocytosis and H. pylori infection suffer from dyspepsia. Duodenal intraepithelial lymphocytosis is significantly associated with bloating (pâ¯=â¯0.0082). CONCLUSIONS: In our cohort of dyspeptic patients, duodenal lymphocytosis is significantly associated with bloating and the simultaneous presence of duodenal lymphocytosis and H. pylori infection is significantly more prevalent than in control subjects.
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Enfermedades Duodenales/patología , Dispepsia/complicaciones , Infecciones por Helicobacter/complicaciones , Helicobacter pylori , Mucosa Intestinal/patología , Linfocitosis/patología , Adulto , Anciano , Anciano de 80 o más Años , Biopsia , Estudios de Casos y Controles , Estudios Transversales , Enfermedades Duodenales/complicaciones , Endoscopía Gastrointestinal , Femenino , Humanos , Linfocitosis/complicaciones , Masculino , Persona de Mediana EdadRESUMEN
BACKGROUND: HLA-G antigens are nonclassical major histocompatibility complex (MHC) class I molecules characterized by tolerogenic and antiinflammatory properties. Recently, a different expression of HLA-G antigens has been observed between intestinal biopsies of ulcerative colitis (UC) and Crohn's disease (CD) patients. These data suggested a functional role for HLA-G molecules in the diseases and proposed the HLA-G modulation as a marker for the diagnosis of UC and CD. The soluble HLA-G antigens (sHLA-G) are circulating molecules mainly produced by activated peripheral blood CD14+ monocytes. METHODS: We tested, by specific enzyme-linked immunosorbent assay (ELISA), the sHLA-G molecule levels in the supernatants of unstimulated and bacterial lipopolysaccharide (LPS)-stimulated cultures of peripheral blood mononuclear cells (PBMC) from 30 healthy subjects, 10 CD, and 18 UC patients. The data were not influenced by treatment or disease activity. RESULTS: The results confirmed a different sHLA-G expression between the diseases, with a spontaneous secretion of sHLA-G in CD patients but not in UC and healthy subjects. Moreover, a lack of sHLA-G antigens has been reported in UC patient cultures after LPS activation but not in healthy subjects and CD patients. The defective sHLA-G production was related to an impaired IL-10 secretion in UC but not in CD. CONCLUSIONS: Overall, these results confirm the presence of a different biological characteristic between CD and UC patients and suggest sHLA-G production by PBMC as a noninvasive diagnostic tool in the early phases of the diseases.
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Colitis Ulcerosa/diagnóstico , Colitis Ulcerosa/inmunología , Enfermedad de Crohn/diagnóstico , Enfermedad de Crohn/inmunología , Antígenos HLA/biosíntesis , Antígenos de Histocompatibilidad Clase I/biosíntesis , Leucocitos Mononucleares/inmunología , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores , Análisis Químico de la Sangre , Ensayo de Inmunoadsorción Enzimática/métodos , Femenino , Antígenos HLA-G , Humanos , Interleucina-10/metabolismo , Masculino , Persona de Mediana EdadRESUMEN
Cerebrospinal fluid (CSF) levels of sHLA-G (sHLA-G1/HLA-G5) molecules and their soluble isoforms HLA-G5 and sHLA-G1 were measured by ELISA procedures in 68 relapsing-remitting Multiple Sclerosis (RR MS) patients, in 67 patients with other inflammatory neurological disorders (OIND) and in 70 subjects with non-inflammatory neurological disorders (NIND). CSF concentrations of sHLA-G1/HLA-G5 and HLA-G5 were higher in RR MS than in OIND and NIND, and in Magnetic Resonance Imaging (MRI) inactive as compared to MRI active RR MS. Our results indicate that the potential implication of sHLA-G proteins in the resolution of MS intrathecal inflammatory response is probably due to HLA-G5 isoform.
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Antígenos HLA/líquido cefalorraquídeo , Antígenos de Histocompatibilidad Clase I/líquido cefalorraquídeo , Esclerosis Múltiple Recurrente-Remitente/líquido cefalorraquídeo , Adulto , Encéfalo/patología , Ensayo de Inmunoadsorción Enzimática/métodos , Femenino , Antígenos HLA-G , Humanos , Imagen por Resonancia Magnética/métodos , Masculino , Persona de Mediana Edad , Esclerosis Múltiple Recurrente-Remitente/patologíaRESUMEN
Human leukocyte antigen-G (HLA-G) is thought to play a key role in implantation by controlling trophoblast invasion and maintaining a local immunosuppressive state. The secretion of soluble HLA-G antigens (sHLA-G) by early embryos seems necessary for a successful implantation and could be a marker of increased pregnancy rate following in vitro fertilization. We have reviewed the results obtained during the last years (from 1987 to 2005). They overall confirmed the predictive role of sHLA-G production in pregnancy outcome. Furthermore, we have examined the technical procedures utilized, with a particular attention to the monoclonal antibodies used in the enzyme-linked immunosorbent assay (ELISA) techniques. New functional roles for HLA-G molecules in pregnancy could be suggested by the relationship observed between the presence of sHLA-G antigens in follicular fluids and sHLA-G expression in the corresponding fertilized oocyte. Furthermore, since maternal mRNA is fundamental for protein production in early embryos, the biologic role of the HLA-G 14 base pair polymorphism could be explored.
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Antígenos HLA/genética , Antígenos de Histocompatibilidad Clase I/genética , Embarazo/inmunología , Femenino , Antígenos HLA/biosíntesis , Antígenos HLA-G , Antígenos de Histocompatibilidad Clase I/biosíntesis , HumanosRESUMEN
The nonclassical human leukocyte antigen (HLA) class Ib gene HLA-G may be important for the induction and maintenance of immune tolerance between the mother and the semi-allogeneic fetus during pregnancy. Expression of HLA-G can influence cytokine and cytotoxic T-lymphocyte responses. Different HLA-G mRNA isoform expression patterns have been associated with HLA-G polymorphism, especially with a 14-bp insertion deletion polymorphism in the 3' untranslated region (3'UTR) of the HLA-G gene. A significantly high level of interleukin-10 (IL-10) secretion is observed in homozygous +14/+14-bp HLA-G peripheral blood mononuclear cells after lipopolysaccharide (LPS) stimulation. This study finds that polymorphism in the 5' upstream regulatory region (5'URR) of the HLA-G gene may also be implicated in differences in IL-10 secretion. However, this may also be due to linkage disequilibrium with the 14-bp polymorphism. A single-nucleotide polymorphism located -477 bp from the start site of exon 1 had a significant association with IL-10 concentrations but not after correction (p=0.011; pc=0.154). This polymorphism is located next to a heat shock element. Eighteen 5'-URR/3'-UTR HLA-G haplotypes were defined; one common homozygous genotype based on these haplotypes was significantly associated with a high IL-10 level after LPS stimulation compared to certain other genotypes. This study indicates that polymorphism in the 5'-URR of the HLA-G gene may have functional significance, although a new line of investigations is needed to elucidate these findings.
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Regiones no Traducidas 3'/genética , Antígenos HLA/genética , Antígenos de Histocompatibilidad Clase I/genética , Interleucina-10/metabolismo , Polimorfismo Genético , Elementos Reguladores de la Transcripción/genética , Secuencia de Bases , Expresión Génica , Antígenos HLA-G , Humanos , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/inmunología , Desequilibrio de Ligamiento , Lipopolisacáridos/farmacología , Datos de Secuencia MolecularRESUMEN
BACKGROUND: Deregulated immune response fails to control biofilm-forming bacteria, as Pseudomonas aeruginosa, in the lungs of cystic fibrosis (CF) patients. HLA-G is an immune-modulatory molecule involved in respiratory diseases and infections. MATERIALS & METHODS: HLA-G mRNA and protein were analyzed in plasma and exhaled breath condensate from CF patients undergoing intravenous antibiotic treatment, CF cell line and murine model. RESULTS: Therapy normalizes HLA-G plasmatic in CF patients suggesting a systemic anti-inflammatory role while in CF airway system, higher expression of HLA-G is associated with P. aeruginosa infection. CF cell line and murine model expressed higher HLA-G molecules in the presence of P. aeruginosa. CONCLUSION: Plasmatic and lung HLA-G expression suggest a role in reducing systemic inflammation and supporting P. aeruginosa infection.
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Fibrosis Quística/complicaciones , Fibrosis Quística/genética , Antígenos HLA-G/genética , Infecciones por Pseudomonas/genética , Infecciones por Pseudomonas/inmunología , Pseudomonas aeruginosa , Administración Intravenosa , Animales , Antibacterianos/administración & dosificación , Antibacterianos/uso terapéutico , Bronquios/inmunología , Líquido del Lavado Bronquioalveolar/inmunología , Línea Celular , Fibrosis Quística/microbiología , Modelos Animales de Enfermedad , Antígenos HLA-G/análisis , Antígenos HLA-G/sangre , Antígenos de Histocompatibilidad Clase I/genética , Interacciones Huésped-Patógeno , Humanos , Inflamación , Pulmón/inmunología , Ratones , Estudios Prospectivos , Infecciones por Pseudomonas/tratamiento farmacológico , Pseudomonas aeruginosa/inmunología , Mucosa Respiratoria/inmunologíaRESUMEN
We have investigated the presence of non-classical soluble HLA-G molecules (sHLA-G) in cerebrospinal fluid (CSF) of multiple sclerosis (MS) patients and the possible relationships between CSF levels of sHLA-G, classical soluble HLA-I (sHLA-I) molecules, IL-10 amounts and Magnetic Resonance Imaging (MRI) findings were evaluated. We studied by ELISA technique the sHLA-I, sHLA-G and IL-10 levels in CSF of 50 relapsing-remitting (RR) MS patients stratified according to clinical and MRI evidence of disease activity. Thirty-six patients with other inflammatory neurological disorders (OIND) and 41 with non-inflammatory neurological disorders (NIND) were used as controls. CSF mean levels were significantly higher in MS and OIND than in NIND for sHLA-I (p<0.001) and in MS than in controls for sHLA-G (p<0.001), with no differences among the various groups for IL-10 mean concentrations. An increase in CSF sHLA-I was found in MS patients with Gd-enhancing lesions (p<0.01), while sHLA-G and IL-10 were more represented in MS patients without lesional activity on MRI scans (p<0.02). In MRI-inactive MS, CSF IL-10 mean concentrations were significantly greater in patients with CSF-detectable levels of sHLA-G than in those without any evidence of CSF sHLA-G expression (p<0.05). Our findings suggest that CSF classical sHLA-I and non-classical sHLA-G levels may modulate MS activity as assessed by MRI acting in opposite directions. The association observed between sHLA-G and IL-10 when Gd-enhancing lesion resolved indicates a potential immunoregulatory role for IL-10 in the control of MS disease activity by shifting the sHLA-I/sHLA-G balance towards sHLA-G response.
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Antígenos HLA/líquido cefalorraquídeo , Antígenos de Histocompatibilidad Clase I/líquido cefalorraquídeo , Interleucina-10/líquido cefalorraquídeo , Imagen por Resonancia Magnética , Esclerosis Múltiple Recurrente-Remitente/líquido cefalorraquídeo , Esclerosis Múltiple Recurrente-Remitente/inmunología , Adulto , Anticuerpos Monoclonales , Western Blotting , Ensayo de Inmunoadsorción Enzimática/estadística & datos numéricos , Femenino , Gadolinio , Antígenos HLA/inmunología , Antígenos HLA-G , Antígenos de Histocompatibilidad Clase I/inmunología , Humanos , Inflamación/líquido cefalorraquídeo , Inflamación/diagnóstico , Inflamación/inmunología , Imagen por Resonancia Magnética/métodos , Imagen por Resonancia Magnética/estadística & datos numéricos , Masculino , Persona de Mediana Edad , Esclerosis Múltiple Recurrente-Remitente/diagnóstico , Enfermedades del Sistema Nervioso/líquido cefalorraquídeo , Enfermedades del Sistema Nervioso/diagnóstico , Enfermedades del Sistema Nervioso/inmunología , SolubilidadRESUMEN
It has recently become clear that interferon-beta (IFN-beta) treatment is effective in ameliorating relapsing-remitting multiple sclerosis (RRMS) through an as yet unidentified mechanism. As there is no recognisable biological indicator to predict responsiveness to IFN-beta treatment, we have investigated fluctuations in serum sHLA-I levels in MS patients undergoing IFN-beta 1b therapy. Serum sHLA-I concentrations measured by enzyme-linked immunosorbent assay (ELISA) were assessed at baseline and, longitudinally, over a period of 18 months after the start of treatment in 29 RRMS patients grouped as responders and nonresponders according to their clinical response to IFN-beta 1b therapy. Thirty-nine healthy volunteers served as controls. Serum sHLA-I concentrations were significantly higher (p<0.001) in pretreated RRMS patients than in healthy donors. In MS patients, changes in mean serum levels of sHLA-I from baseline showed a temporal pattern characterized by a strong increase in the first trimester of treatment, a return toward basal values in the following 6 months, a slight decline at 12th and 15th months and a further moderate increase at the 18th month. Mean serum sHLA-I levels were significantly more elevated in responders than in nonresponders at the first (p<0.02), second (p<0.01), and at third (p<0.02) months after the beginning of treatment and significantly lower (p<0.01) at the time of relapses in comparison to baseline values. Overall, these results seem to indicate that IFN-beta 1b can modulate fluctuations in serum sHLA-I levels and argue in favour of a potential role for serum levels of sHLA-I as a sensitive marker to monitor response to IFN-beta treatment in MS.
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Antígenos HLA/sangre , Interferón beta/uso terapéutico , Esclerosis Múltiple Recurrente-Remitente/tratamiento farmacológico , Adulto , Antineoplásicos/uso terapéutico , Evaluación de la Discapacidad , Ensayo de Inmunoadsorción Enzimática/métodos , Femenino , Humanos , Interferon beta-1b , Estudios Longitudinales , Masculino , Persona de Mediana Edad , Esclerosis Múltiple Recurrente-Remitente/sangre , Estadísticas no Paramétricas , Factores de TiempoRESUMEN
The goal of our study was to clarify the contribution of soluble human leukocyte antigens class I (sHLA-I) in multiple sclerosis (MS) immune dysregulation. We retrospectively evaluated by ELISA cerebrospinal fluid (CSF) and serum sHLA-I levels in 79 relapsing-remitting (RR), 26 secondary progressive (SP) and 15 primary progressive (PP) MS patients stratified according to clinical and Magnetic Resonance Imaging (MRI) evidence of disease activity. One hundred and nine patients with other inflammatory neurological disorders (OIND), 88 with noninflammatory neurological disorders (NIND) and 82 healthy donors were used as controls. An intrathecal synthesis of sHLA-I detected by a specific index was significantly more consistent in MS than in controls, with more pronounced values in MS patients with relapses and MRI enhancing brain lesions. A decrease in serum sHLA-I concentrations was observed in MS patients with demyelinating attacks, while an increase in CSF levels of sHLA-I was found in MS patients with lesional activity on MRI scans. This association between intrathecal synthesis and reciprocal fluctuations of CSF and serum levels of sHLA-I in clinically and MRI active MS seems to suggest a potential role for CSF and serum levels of sHLA-I as a sensitive marker of immune activation taking place both intrathecally and systemically in MS.
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Sistema Nervioso Central/inmunología , Antígenos de Histocompatibilidad Clase I/sangre , Antígenos de Histocompatibilidad Clase I/líquido cefalorraquídeo , Esclerosis Múltiple/sangre , Esclerosis Múltiple/líquido cefalorraquídeo , Adulto , Sistema Nervioso Central/metabolismo , Sistema Nervioso Central/patología , Progresión de la Enfermedad , Femenino , Humanos , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Esclerosis Múltiple/patología , Valor Predictivo de las Pruebas , Estudios RetrospectivosRESUMEN
The current management of early rheumatoid arthritis (ERA) is to start an intensive treatment as soon as possible. To avoid under/overtreatment, it is important to identify reliable ERA evolution biomarkers. HLA-G molecules has been associated with rheumatoid arthritis, suggesting a role in disease regulation. HLA-G antigens are expressed as membrane bound and soluble isoforms (mHLA-G, sHLA-G) that act as ligand for immune-inhibitory receptors (ILT2, ILT4, KIR2DL4). Expression of HLA-G is influenced by a 14 bp insertion/deletion polymorphism in exon 8 of the gene, where the deletion is associated with mRNA stability. We analyzed 23 ERA patients during a 12 months follow-up disease treatment for sHLA-G, IL-1beta, IL-6, IL-10 and TNF-alpha levels in plasma samples by ELISA, mHLA-G and ILT2 expression on peripheral blood CD14 positive cells by flow cytometry and typed HLA-G 14 bp deletion/insertion polymorphism by Real-Time PCR. Disease status (DAS28), ultrasonography with power Doppler and laboratory data were checked. Cytokine levels confirmed the anti-inflammatory effect of the treatment. sHLA-G, mHLA-G and ILT2 expression inversely correlated with DAS28 disease scores. The frequency of 14 bp deletion allele increased in patients with disease remission. Based on these results, HLA-G may be a candidate biomarker to evaluate early prognosis and disease activity in ERA patients.
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Artritis Reumatoide/sangre , Artritis Reumatoide/diagnóstico , Antígenos HLA-G/inmunología , Mutación , Anciano , Alelos , Antígenos CD/sangre , Antígenos CD/inmunología , Artritis Reumatoide/genética , Artritis Reumatoide/inmunología , Biomarcadores/sangre , Citocinas/sangre , Citocinas/inmunología , Análisis Mutacional de ADN , Diagnóstico Precoz , Exones , Femenino , Expresión Génica , Antígenos HLA-G/sangre , Humanos , Receptor Leucocitario Tipo Inmunoglobulina B1 , Masculino , Persona de Mediana Edad , Pronóstico , Isoformas de Proteínas/sangre , Isoformas de Proteínas/inmunología , Receptores Inmunológicos/sangre , Receptores Inmunológicos/inmunologíaRESUMEN
Human umbilical cord blood units (UCBs) are an alternative source in allogeneic-stem-cell transplantation. Human leukocyte antigen (HLA)-G is a tolerogenic molecule with a possible implication in UCB immunoregulatory effect. HLA-G expression was observed in UCB myeloid and plasmacytoid dendritic cells; in contrast, CD34(+) cells did not produce this molecule. CD34(+) cells are primitive hematopoietic progenitor cells that are present in UCB and are necessary for long-term engraftment via production of immunoregulatory molecules and a hematopoietic progeny that supports cellular recovery. The role of these cells in UCB transplantation needs further evaluation of HLA-G expression in CD34(+) cells and their hematopoietic progeny. We confirmed the absence of HLA-G expression in CD34(+) cells, whereas CD34(+)-derived progeny secreted HLA-G molecules and expressed HLA-G mRNA in in vitro cultures. Furthermore, soluble HLA (sHLA)-G molecules purified from the culture supernatants of CD34(+)-derived progeny were able to suppress lymphoproliferative response in an HLA-G dose-dependent manner. Overall these results identify CD34(+)-derived hematopoietic progeny as producers of HLA-G molecules and support a role of this antigen as an immuno-modulatory factor in UCB.
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Antígenos CD34/inmunología , Sangre Fetal/citología , Sangre Fetal/inmunología , Antígenos HLA-G/inmunología , Adulto , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , Femenino , Regulación de la Expresión Génica , Antígenos HLA-G/aislamiento & purificación , Antígenos HLA-G/farmacología , Humanos , Factores Inmunológicos/farmacología , EmbarazoRESUMEN
HLA-G is believed to act as an anti-inflammatory molecule in Multiple Sclerosis (MS). The 3' untranslated region of the HLA-G gene is characterized by two polymorphisms, DEL/INS14bp and +3142C>G, which control soluble HLA-G (sHLA-G) production. The influence of these two HLA-G variants on sHLA-G serum and cerebrospinal fluid (CSF) levels was investigated in 69 Relapsing-Remitting MS patients grouped in magnetic resonance imaging (MRI) inactive and active disease. Serum and CSF sHLA-G levels were more elevated in high than in low DEL/INS 14bp and +3142C>G sHLA-G producers and were different among the various combined HLA-G genotypes in both MRI inactive and active diseases. The highest and the lowest sHLA-G values were identified in MS patients with C/C,DEL/DEL and G/G,INS/INS genotypes, respectively. Our preliminary findings suggest that serum and CSF sHLA-G levels in MS could be influenced by HLA-G polymorphisms irrespective of the inflammatory microenvironment.
Asunto(s)
Antígenos HLA-G/genética , Esclerosis Múltiple Recurrente-Remitente/genética , Esclerosis Múltiple Recurrente-Remitente/metabolismo , Mutagénesis Insercional , Polimorfismo de Nucleótido Simple , Eliminación de Secuencia , Adulto , Alelos , Femenino , Frecuencia de los Genes , Genotipo , Antígenos HLA-G/sangre , Antígenos HLA-G/líquido cefalorraquídeo , Humanos , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Esclerosis Múltiple Recurrente-Remitente/diagnósticoRESUMEN
CONTEXT: Type 2 diabetes mellitus (T2DM) and obesity are characterized by a low-grade inflammation, which might be related to the development of insulin resistance. Human leukocyte antigen-G (HLA-G) shows antiinflammatory and tolerogenic properties, including the modulation of CD8+ T-cell cytotoxicity and regulation of CD4+ T-lymphocyte function. These functions are partially shared with IL-10, whose levels are reduced in insulin-resistant states. OBJECTIVE: The aim was to explore the relationship between HLA-G and the metabolic and inflammatory pattern of obesity or T2DM. PATIENTS AND MAIN OUTCOME MEASURES: Soluble HLA-G, IL-6, and IL-10 were measured and related with metabolic and biochemical parameters in 230 volunteers with normal glucose tolerance, impaired glucose tolerance, or T2DM by oral glucose tolerance test. RESULTS: sHLA-G, detected in 144 subjects (sHLA-G positive), was more frequent in T2DM or impaired glucose tolerance subjects than in normal glucose tolerance (chi(2) =18.6; P < 0.0001), and its plasma levels increased progressively across the classes of glucose tolerance. sHLA-G-positive individuals had higher body mass index, systolic blood pressure, and cholesterol levels; a reduced degree of insulin sensitivity; and almost 2-fold higher levels of IL-6, a cytokine related to insulin sensitivity, whereas IL-10 was similar. In the sHLA-G-positive subgroup, by a multivariate regression model, sHLA-G was significantly related to 2-h glucose, the area under insulin curve, and IL-6 levels (multiple r(2) = 0.14; P < 0.001), independently of age, gender, and body mass index. CONCLUSIONS: A frequent expression of sHLA-G, linked to a typical biomarker of insulin resistance like IL-6, seems to characterize subjects with an impaired glucose metabolism.