RESUMEN
A Dengue induz uma resposta exacerbada e transitória das células secretoras de anticorpos (ASCs) no sangue de pacientes cerca de sete dias após o início dos sintomas. A frequência dessas ASCs chega a representar mais de 50% de todas as células B circulantes neste período. No entanto, ainda é desconhecido se a magnitude dessa resposta tem relação com a gravidade da Dengue. Nosso grupo de pesquisa já mostrou que a cultura de células nucleares do sangue periférico (PBMCs) de indivíduos saudáveis com partículas do vírus da Dengue (DENV), por 7 dias, levava a diferenciação de células B em ASCs em magnitude similar àquelas estimuladas por mitógenos. Essas culturas apresentavam um consumo significativamente maior de triptofano (TRP), associado à maior expressão das enzimas IDO1 e IDO2, e, consequentemente, maior síntese de quinurenina (KYN) em relação ao estímulo por mitógenos. Considerando que as concentrações de TRP e KYN detectadas nos sobrenadantes dessas culturas eram diretamente proporcionais ao aumento de ASCs, decidimos investigar o papel desse metabolismo do TRP e de seus respectivos metabólitos na diferenciação das ASCs. Para isso, análises do transcriptoma público de células únicas do sangue periférico de pacientes com Dengue (estudo E-MTAB- 9467) foram realizadas para inferir a real participação do metabolismo do TRP na geração de ASCs. Com o programa R foram executadas análises de Downstream. Identificamos um aumento massivo das ASCs nas amostras dos pacientes infectados com Dengue. No entanto, os principais genes desencadeadores da ativação do metabolismo do TRP (IDO1 e IDO2) não foram expressos nas subpopulações de células B, mas sim em células dendríticas e monócitos CD14+ respectivamente. Isso sugeriria que esta via não seria ativada nos linfócitos B. Por outro lado, genes codificadores de outros participantes da via do TRP (HSD17B10, ECHS1 e SIRT3) foram detectados em células B e podem estar relacionados com a proliferação das ASCs. Além disso, a análise de enriquecimento mostrou uma aumentada expressão de genes associados com moléculas de MHC de classe II em plasmablastos e plasmócitos de pacientes com Dengue. Porém, com a expressão aumentada de ENTPD1 nessas células durante a fase sintomatológica, nossos dados sugerem também que um eventual papel de plasmablastos e plasmócitos como apresentadoras de antígenos na Dengue poderia induzir uma resposta supressora de células T
Dengue can cause an exacerbated and transient antibody-secreting cell (ASC) response in the blood of patients nearly 7 days of symptomatology. The ASC frequency reaches more than 50% of all circulating B cells during that period. However, it is still unknown whether the magnitude of this response may be directly related to the severity of Dengue. Our research group has already shown that the culture of peripheral blood mononuclear cells (PBMCs) from healthy individuals with Dengue virus (DENV) particles for 7 days led to a differentiation of B cells into ASCs to a magnitude similar to those stimulated by mitogens. These cultures showed significantly higher consumption of tryptophan (TRP), associated with higher expression of enzymes IDO1 and IDO2, and consequently, higher synthesis of quinurenine (KYN) compared to mitogen stimulation. The concentrations of TRP and KYN detected in the supernatants of these cultures were directly proportional to ASC frequency increase. Thus, we have decided to investigate the role of TRP metabolism and its respective metabolites in ASC differentiation. For this, we performed an analysis of single-cell transcriptome with peripheral blood from Dengue patients (dataset from E-MTAB-9467 study). Downstream analyses were performed with R software. Corroborating with literature, we identified a massive increase in ASC frequency of Dengue infected patients. However, the main genes triggering TRP metabolism activation (IDO1 and IDO2) were not expressed in B-cell subsets, but in dendritic cells and CD14+ monocytes, respectively. This would suggest that this pathway would not be activated in B lymphocytes. Nevertheless, genes encoding other participants in the TRP pathway (HSD17B10, ECHS1, and SIRT3) were detected in B cells and may be related to ASC proliferation. Futhermore, a Gene Ontology analysis showed an increased expression of genes associated with MHC class II molecules in plasmablasts and plasma cells of Dengue patients. As these cells also presented an increased expression of ENTPD1 during the symptomatic phase, our data suggest a potential role of plasmablasts and plasma cells as antigen-presenting cells associated with a suppressive T cell response in Dengue
Asunto(s)
Pacientes/clasificación , Células Plasmáticas/clasificación , Antígenos de Histocompatibilidad Clase II/farmacología , Dengue/patología , Análisis de Expresión Génica de una Sola Célula/instrumentación , Triptófano/análogos & derivadosRESUMEN
The increasingly recognized role of different types of B cells and plasma cells in protective and pathogenic immune responses combined with technological advances have generated a plethora of information regarding the heterogeneity of this human immune compartment. Unfortunately, the lack of a consistent classification of human B cells also creates significant imprecision on the adjudication of different phenotypes to well-defined populations. Additional confusion in the field stems from: the use of non-discriminatory, overlapping markers to define some populations, the extrapolation of mouse concepts to humans, and the assignation of functional significance to populations often defined by insufficient surface markers. In this review, we shall discuss the current understanding of human B cell heterogeneity and define major parental populations and associated subsets while discussing their functional significance. We shall also identify current challenges and opportunities. It stands to reason that a unified approach will not only permit comparison of separate studies but also improve our ability to define deviations from normative values and to create a clean framework for the identification, functional significance, and disease association with new populations.
Asunto(s)
Linfocitos B Reguladores/inmunología , Linfocitos B/inmunología , Memoria Inmunológica/inmunología , Células Plasmáticas/inmunología , Animales , Linfocitos B/clasificación , Linfocitos B Reguladores/metabolismo , Citocinas/inmunología , Citocinas/metabolismo , Humanos , Inmunidad Humoral/inmunología , Inmunoglobulina D/inmunología , Inmunoglobulina D/metabolismo , Inmunoglobulina M/inmunología , Inmunoglobulina M/metabolismo , Células Plasmáticas/clasificaciónRESUMEN
Plasma cell segmentation is the first stage of a computer assisted automated diagnostic tool for multiple myeloma (MM). Owing to large variability in biological cell types, a method for one cell type cannot be applied directly on the other cell types. In this paper, we present PCSeg Tool for plasma cell segmentation from microscopic medical images. These images were captured from bone marrow aspirate slides of patients with MM. PCSeg has a robust pipeline consisting of a pre-processing step, the proposed modified multiphase level set method followed by post-processing steps including the watershed and circular Hough transform to segment clusters of cells of interest and to remove unwanted cells. Our modified level set method utilizes prior information about the probability densities of regions of interest (ROIs) in the color spaces and provides a solution to the minimal-partition problem to segment ROIs in one of the level sets of a two-phase level set formulation. PCSeg tool is tested on a number of microscopic images and provides good segmentation results on single cells as well as efficient segmentation of plasma cell clusters.
Asunto(s)
Diagnóstico por Computador/métodos , Mieloma Múltiple/diagnóstico , Células Plasmáticas/clasificación , Algoritmos , Humanos , Procesamiento de Imagen Asistido por Computador , Microscopía , Mieloma Múltiple/patología , Reconocimiento de Normas Patrones Automatizadas/métodos , Células Plasmáticas/citología , Programas InformáticosRESUMEN
Plasma cells are rare cells that have been notoriously difficult to detect by flow cytometry. New advances have described B220+ CD138+ plasma cells in the bone marrow that are particularly difficult to distinguish between CD138 intermediate B220+ developing B cells. Herein we describe a novel method for detecting plasma cells in the bone marrow using a combination of CD138 and Sca-1 staining.
Asunto(s)
Antígenos Ly/análisis , Citometría de Flujo/métodos , Inmunofenotipificación/métodos , Proteínas de la Membrana/análisis , Células Plasmáticas/clasificación , Células Plasmáticas/inmunología , Sindecano-1/análisis , Animales , Células de la Médula Ósea/inmunología , Antígenos Comunes de Leucocito/análisis , Ratones , Ganglios Linfáticos Agregados/citología , Ganglios Linfáticos Agregados/inmunología , Factor 1 de Unión al Dominio 1 de Regulación Positiva , Bazo/citología , Bazo/inmunología , Factores de Transcripción/análisisRESUMEN
We provide a robust four-color fluorescence-based flow cytometry protocol that distinguishes viable dividing plasmablasts from nondividing plasma cells and, based on CD19 surface abundance, identifies two mature plasma cell populations in the spleen and the bone marrow of mice.
Asunto(s)
Citometría de Flujo/métodos , Inmunofenotipificación/métodos , Células Plasmáticas/clasificación , Células Plasmáticas/inmunología , Coloración y Etiquetado/métodos , Animales , Antígenos CD19/análisis , Células de la Médula Ósea/inmunología , Diferenciación Celular , Proteínas Fluorescentes Verdes , Inmunofenotipificación/instrumentación , Antígenos Comunes de Leucocito/análisis , Ratones , Factor 1 de Unión al Dominio 1 de Regulación Positiva , Bazo/inmunología , Sindecano-1/análisis , Factores de Transcripción/análisisRESUMEN
OBJECTIVE: To study the clinical, pathologic, immunophenotype, molecular characteristics and prognosis of HIV-negative plasmablastic lymphoma (PBL). METHODS: Twelve cases of HIV-negative PBLs diagnosed between 2005 and 2014 in Guangdong General Hospital were identified according to WHO classification of tumors of haematopoietic and lymphoid tissues (2008). The clinicopathologic features and outcome were analyzed and the relevant literatures were reviewed. RESULTS: The patients were predominantly male (11/12) with a median age of 55.5 years. The tumor cells showed the characteristic combination of immunoblastic/plasmablastic morphology, plasma cell phenotype and high proliferation, no expression of mature B cell markers. 7/10 of the cases were EBER positive. Two cases were positive for C-myc translocation. Four of twelve patients were died. CONCLUSIONS: PBL is a rare, aggressive B-cell lymphoma. HIV-negative PBL has lower rate of oral involvement and EBER expression than HIV-positive patients, the differential diagnosis is very challenging, and the prognosis is worse.
Asunto(s)
Seronegatividad para VIH , Linfoma Plasmablástico/diagnóstico , Linfoma Plasmablástico/patología , Diagnóstico Diferencial , Femenino , Humanos , Inmunofenotipificación , Linfoma de Células B/diagnóstico , Masculino , Persona de Mediana Edad , Células Plasmáticas/clasificación , Pronóstico , Translocación GenéticaRESUMEN
Normal and reactive plasma cells (PC) are easy to ascertain on human bone marrow films, due to their small mature-appearing nucleus and large cytoplasm, the latter usually deep blue after Giemsa staining. Cytoplasm is filled with long strands of rough endoplasmic reticulum and one large Golgi apparatus (paranuclear hof), demonstrating that PC are dedicated mainly to protein synthesis and excretion (immunoglobulin). Deregulation of the genome may induce clonal expansion of one PC that will lead to immunoglobulin overproduction and eventually to one among the so-called PC neoplasms. In multiple myeloma (MM), the number of PC is over 10% in most patients studied. Changes in the morphology of myeloma PC may be inconspicuous as compared to normal PC (30-50% patients). In other instances PC show one or several morphological changes. One is related to low amount of cytoplasm, defining lymphoplasmacytoid myeloma (10-15% patients). In other cases (40-50% patients), named immature myeloma cases, nuclear-cytoplasmic asynchrony is observed: presence of one nucleolus, finely dispersed chromatin and/or irregular nuclear contour contrast with a still large and blue (mature) cytoplasm. A peculiar morphological change, corresponding to the presence of very immature PC named plasmablasts, is observed in 10-15% cases. Several prognostic morphological classifications have been published, as mature myeloma is related to favorable outcome and immature myeloma, peculiarly plasmablastic myeloma, is related to dismal prognosis. However, such classifications are no longer included in current prognostic schemes. Changes related to the nucleus are very rare in monoclonal gammopathy of unknown significance (MGUS). In contrast, anomalies related to the cytoplasm of PC, including color (flaming cells), round inclusions (Mott cells, Russell bodies), Auer rod-like or crystalline inclusions, are reported in myeloma cases as well as in MGUS and at times in reactive disorders. They do not correspond to malignant changes of PC but are related to abnormal synthesis, trafficking, or excretion of the immunoglobulin that is stored in excess within the cytoplasm. Occurrence of crystalline inclusions within PC may be the first anomaly leading to the diagnosis of adult Fanconi syndrome. After a historical perspective, the authors report on the various morphological aspects of PC that may occur in multiple myeloma and related disorders, and discuss about their clinical and pathophysiological significance. Today, morphological identification and accurate determination of % PC within bone marrow remain ancillary criteria for the diagnosis of MM and help for the diagnosis of rare renal disorders.
Asunto(s)
Mieloma Múltiple/patología , Células Plasmáticas/patología , Médula Ósea/patología , Recuento de Células , Forma de la Célula , Crioglobulinemia/patología , Citoplasma/ultraestructura , Síndrome de Fanconi/patología , Citometría de Flujo , Histiocitosis/patología , Humanos , Cuerpos de Inclusión/ultraestructura , Hierro/análisis , Leucemia de Células Plasmáticas/patología , Gammopatía Monoclonal de Relevancia Indeterminada/patología , Mieloma Múltiple/diagnóstico , Proteínas de Mieloma/análisis , Células Madre Neoplásicas/patología , Orgánulos/ultraestructura , Células Plasmáticas/clasificación , Pronóstico , Coloración y EtiquetadoRESUMEN
Specific serum antibodies mediating humoral immunity and autoimmunity are provided by mature plasma cells (PC) residing in the bone marrow (BM), yet their dynamics and composition are largely unclear. We here characterize distinct subsets of human PC differing by CD19 expression. Unlike CD19(+) PC, CD19(-) PC were restricted to BM, expressed predominantly IgG, and they carried a prosurvival, distinctly mature phenotype, that is, HLA-DR(low)Ki-67(-)CD95(low)CD28(+)CD56(+/-), with increased BCL2 and they resisted their mobilization from the BM after systemic vaccination. Fewer mutations within immunoglobulin VH rearrangements of CD19(-) BMPC may indicate their differentiation in early life. Their resistance to in vivo B-cell depletion, that is, their independency from supply with new plasmablasts, is consistent with long-term stability of this PC subset in the BM. Moreover, CD19(-) PC were detectable in chronically inflamed tissues and secreted autoantibodies. We propose a multilayer model of PC memory in which CD19(+) and CD19(-) PC represent dynamic and static components, respectively, permitting both adaptation and stability of humoral immune protection.
Asunto(s)
Antígenos CD19/metabolismo , Células de la Médula Ósea/citología , Células de la Médula Ósea/inmunología , Inmunoglobulina G/metabolismo , Células Plasmáticas/inmunología , Suero Antilinfocítico/administración & dosificación , Células de la Médula Ósea/clasificación , Ácidos Borónicos/administración & dosificación , Bortezomib , Diferenciación Celular , Supervivencia Celular , Vacuna contra Difteria y Tétanos/administración & dosificación , Humanos , Inmunidad Humoral , Memoria Inmunológica , Inflamación/inmunología , Inflamación/patología , Depleción Linfocítica , Modelos Inmunológicos , Mutación , Fenotipo , Células Plasmáticas/clasificación , Células Plasmáticas/citología , Pirazinas/administración & dosificación , Recombinación V(D)JRESUMEN
Circulating myeloma tumor cells (CTCs) as defined by the presence of peripheral blood (PB) clonal plasma cells (PCs) are a powerful prognostic marker in multiple myeloma (MM). However, the biological features of CTCs and their pathophysiological role in MM remains unexplored. Here, we investigate the phenotypic, cytogenetic, and functional characteristics as well as the circadian distribution of CTCs vs paired bone marrow (BM) clonal PCs from MM patients. Our results show that CTCs typically represent a unique subpopulation of all BM clonal PCs, characterized by downregulation (P < .05) of integrins (CD11a/CD11c/CD29/CD49d/CD49e), adhesion (CD33/CD56/CD117/CD138), and activation molecules (CD28/CD38/CD81). Fluorescence in situ hybridization analysis of fluorescence-activated cell sorter-sorted CTCs also unraveled different cytogenetic profiles vs paired BM clonal PCs. Moreover, CTCs were mostly quiescent and associated with higher clonogenic potential when cocultured with BM stromal cells. Most interestingly, CTCs showed a circadian distribution which fluctuates in a similar pattern to that of CD34(+) cells, and opposite to stromal cell-derived factor 1 plasma levels and corresponding surface expression of CXC chemokine receptor 4 on clonal PCs, suggesting that in MM, CTCs may egress to PB to colonize/metastasize other sites in the BM during the patients' resting period.
Asunto(s)
Mieloma Múltiple/sangre , Células Neoplásicas Circulantes/patología , Antígenos CD/sangre , Ciclo Celular , Ritmo Circadiano , Análisis Citogenético , Humanos , Inmunofenotipificación , Mieloma Múltiple/genética , Mieloma Múltiple/inmunología , Células Neoplásicas Circulantes/clasificación , Células Neoplásicas Circulantes/inmunología , Células Plasmáticas/clasificación , Células Plasmáticas/inmunología , Células Plasmáticas/patología , Pronóstico , Estudios Prospectivos , Ensayo de Tumor de Célula MadreRESUMEN
Bone marrow flow cytometric analysis is a powerful and rapid tool for evaluating aberrant plasma cell. In this study, we have examined the utility of multiparameter flow cytometry (MFC) in 52 patients with multiple myeloma (MM) and in 45 patients with monoclonal gammopathy with unknown significance (MGUS) into routine evaluation for the management of patients with plasma cell-related disorders. The plasma cells (PC) were identified by their light scatter distribution and reactivity patterns to CD138, CD38, and CD45. The combination of these parameters was helpful for identifying distinct subpopulations of PCs. Moderate to bright expression of CD56, CD20, CD24, CD28, and CD117 was detected in 67%, 26%, 13%, 27%, and 57% of MM cases and in 58%, 20%, 11%, 43% and 44% of MGUS cases, respectively. In MGUS group, the median percentage abnormal PCs/total PCs was 88% with 37 patients out of 45 (82%) with ratio <95%. The median ratio of the MM group was 98.9% and a ratio ≥ 95% was observed in 37 samples out of 44 (84%). In conclusion, MFC immunophenotyping of PCs has obvious clinical relevance in differential diagnosis between MM and others monoclonal gammopathies, identification of high-risk MGUS and smouldering MM, and minimal residual disease monitoring of MM. Our results showed that this tool can be easily applied in haematology laboratories.
Asunto(s)
Citometría de Flujo/métodos , Inmunofenotipificación/métodos , Gammopatía Monoclonal de Relevancia Indeterminada/diagnóstico , Gammopatía Monoclonal de Relevancia Indeterminada/terapia , Células Plasmáticas/clasificación , Adulto , Anciano , Anciano de 80 o más Años , Estudios de Cohortes , Color , Estudios de Factibilidad , Femenino , Humanos , Masculino , Persona de Mediana Edad , Gammopatía Monoclonal de Relevancia Indeterminada/inmunología , Gammopatía Monoclonal de Relevancia Indeterminada/patología , Mieloma Múltiple/diagnóstico , Mieloma Múltiple/inmunología , Mieloma Múltiple/patología , Valor Predictivo de las Pruebas , PronósticoRESUMEN
While endometrial neutrophils and plasma cells are criteria used to diagnose histologic endometritis in epidemiologic pelvic inflammatory disease (PID) research, plasma cell misidentification and nonspecificity may limit the accuracy of these criteria. Herein, we examined: (1) the identification of endometrial plasma cells with conventional methyl green pyronin-based methodology versus plasma cell-specific (CD138) immunostaining, (2) the prevalence of endometrial plasma cells among women at low risk for PID, and (3) endometrial leukocyte subpopulations among women diagnosed with acute or chronic histologic endometritis by conventional criteria. We observed an absence of CD138+ cells in 25% of endometrial biopsies in which plasma cells had been identified by conventional methodology, while additional immunohistochemical analyses revealed indistinguishable inflammatory infiltrates among women diagnosed with acute or chronic endometritis by conventional criteria. Among women considered at lower risk for PID development, flow cytometric analyses detected plasma cells in 30% of endometrial biopsy specimens, suggesting that these cells, even when accurately identified, only nonspecifically identify upper genital tract inflammatory processes. Combined, our findings underscore the limitations of the criteria used to diagnose histologic endometritis in PID-related research and suggest that satisfactory understanding of PID pathogenesis, treatment, and prevention is hindered by continued use of these criteria.
Asunto(s)
Endometritis/diagnóstico , Endometrio/patología , Neutrófilos/patología , Enfermedad Inflamatoria Pélvica/diagnóstico , Células Plasmáticas/clasificación , Sindecano-1/metabolismo , Adolescente , Adulto , Anticuerpos Monoclonales , Biopsia , Endometritis/epidemiología , Estudios Epidemiológicos , Femenino , Citometría de Flujo , Humanos , Inmunohistoquímica , Leucocitos/citología , Enfermedad Inflamatoria Pélvica/epidemiología , Células Plasmáticas/patología , Valor Predictivo de las Pruebas , Prevalencia , Sensibilidad y Especificidad , Adulto JovenRESUMEN
Multiple myeloma (MM) is a hematological malignancy caused by clonal proliferation of malignant plasma cells (PC). The aim of the work is to determine prognostic significance of morphological subtypes of PC in relation to overall treatment response, long-term survival and other conventional prognostic parameters. One hundred and thirty-nine newly diagnosed MM patients who underwent autologous transplantation in clinical trials conducted in one center were included. Percentual representation of subtypes of plasma cells in bone marrow was measured based on progressive analysis of nucleolus, nuclear chromatin and ratio of nuclei to the volume of cytoplasm (N/C ratio) creating 8 subtypes P000-P111 and four subclassifications of cells. Mature plasma cells (P000, P001) were found in 42.4% of patients; proplasmocytes I (P010, P011, P100) in 38.1% of patients, and proplasmocytes II (P101, P110) in 19.4% of patients. Patients who reached treatment response after autologous transplantation had statistically significant lower frequency of mature plasma cells than patients with no treatment response (median 24.0% vs. 36.0 %; p=0.032). Patients with mature plasma cells of subtype P000 an patients with value P000 ≥ 37% (median 46.8 months vs. 77.8 months; p = 0.020). Patients with proplasmocytes II subtype P110 rings valuable prognostic information and correlation with other prognostic factors as well as total treatment response and survival in MM patients who underwent autologous transplantation.
Asunto(s)
Mieloma Múltiple/mortalidad , Mieloma Múltiple/patología , Células Plasmáticas/clasificación , Células Plasmáticas/patología , Trasplante de Médula Ósea , Nucléolo Celular/genética , Núcleo Celular/genética , Cromatina/genética , Citoplasma/metabolismo , Progresión de la Enfermedad , Femenino , Humanos , Hibridación Fluorescente in Situ , Masculino , Persona de Mediana Edad , Mieloma Múltiple/terapia , Pronóstico , Tasa de Supervivencia , Factores de Tiempo , Trasplante AutólogoRESUMEN
The technological development of flow cytometry (FC) together with new findings reveal the need for immunophenotyping in research of monoclonal gammopathy (MG) because of its diagnostic, prognostic and predictive significance. The aim of the European Myeloma Network (EMN) is to standardize this analytical method and implement it into routine clinical examination. Since the overall significance and application of FC are still analysed, standardisation could help obtain more clinical relevant information in terms of MG pathophysiology.
Asunto(s)
Citometría de Flujo , Paraproteinemias/inmunología , Células Plasmáticas/inmunología , Antígenos CD/análisis , Citometría de Flujo/métodos , Citometría de Flujo/normas , Humanos , Inmunofenotipificación , Paraproteinemias/patología , Células Plasmáticas/clasificaciónRESUMEN
In this paper, initial processing of biological material, cell separation algorithms and other procedures are discussed. For samples with initial infiltration of plasma cells > 5%, CD138 MicroBeads and Auto-Magnetic-Activated Cell Sorting program are used. Fluorescence-Activated Cell Sorting is used exclusively for cell populations with low-abundance; these samples are detected using fluorescently labeled antibodies only. Isolated plasma cells are further processed for molecular biological studies, for cytogenetics and protein analyses. Furthermore, this work examines the pitfalls of research related to multiple myeloma; some of them we have overcome, while the others are still problematic.
Asunto(s)
Citometría de Flujo/métodos , Mieloma Múltiple/diagnóstico , Bancos de Muestras Biológicas , Investigación Biomédica , Médula Ósea/patología , Humanos , Células Plasmáticas/clasificación , Células Plasmáticas/inmunología , Manejo de Especímenes , Sindecano-1/análisisRESUMEN
OBJECTIVES: Castleman's disease (CD) is a group of rare atypical lymphoproliferative disorders classified as hyaline vascular (HV) and plasma cell (PC) types. CD may be closely mimicked by IgG4-related sclerosing disease (IgG4-SD) involving the lymph nodes. We retrospectively analyzed findings in patients with CD to elucidate the relationship between CD and IgG4-SD. METHODS: Clinicopathological and immunophenotypical characteristics, including IgG+ and IgG4 expression by plasma cells, were analyzed in 87 consecutive patients diagnosed with CD from 1999 to 2010 at two major Korean hospitals. RESULTS: The numbers of IgG+ (p < 0.001) and IgG4+ (p < 0.001) cells and the IgG4:IgG ratio (p = 0.003) were significantly higher in the PC than in the HV group. The mean IgG4+:IgG+ plasma cell ratio in the PC group was 25.1%, with 10 patients having a ratio >40%, the threshold IgG4:IgG ratio in patients with IgG4-SD. CONCLUSIONS: Patients with the PC form of CD and IgG4-related lymphadenopathy share some features, including the pattern of plasma cell distribution and high-level infiltration of IgG4+ cells. Some patients thought to have the PC form of CD could be reclassified as showing IgG4-related lymphadenopathy.
Asunto(s)
Enfermedad de Castleman/inmunología , Enfermedad de Castleman/patología , Inmunoglobulina G/metabolismo , Enfermedades Linfáticas/inmunología , Enfermedades Linfáticas/patología , Células Plasmáticas/patología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Enfermedad de Castleman/clasificación , Enfermedad de Castleman/diagnóstico , Niño , Diagnóstico Diferencial , Femenino , Humanos , Inmunohistoquímica , Enfermedades Linfáticas/diagnóstico , Masculino , Persona de Mediana Edad , Células Plasmáticas/clasificación , Células Plasmáticas/inmunología , Estudios Retrospectivos , Esclerosis , Adulto JovenRESUMEN
Sustained long-term antibody levels are the cornerstone of protective immunity, yet it remains unclear how they are durably maintained. A predominant theory implicates antigen-independent antibody production by a subset of long-lived plasma cells (LLPCs) that survive within bone marrow (BM). Central tenets of this model--that BM LLPCs constitute a subset defined by intrinsic biology distinct from PCs in other tissues and contribute to long-term antibody titers--have not been definitively demonstrated. We now report that long-term humoral immunity depends on the PC-intrinsic function of CD28, which selectively supports the survival of BM LLPC but not splenic short-lived PC (SLPC). LLPC and SLPC both express CD28, but CD28-driven enhanced survival occurred only in the LLPC. In vivo, even in the presence of sufficient T cell help, loss of CD28 or its ligands CD80 and CD86 caused significant loss of the LLPC population, reduction of LLPC half-life from 426 to 63 d, and inability to maintain long-term antibody titers, but there was no effect on SLPC populations. These findings establish the existence of the distinct BM LLPC subset necessary to sustain antibody titers and uncover a central role for CD28 function in the longevity of PCs and humoral immunity.
Asunto(s)
Formación de Anticuerpos , Antígenos CD28/metabolismo , Células Plasmáticas/inmunología , Animales , Antígeno B7-1/genética , Antígeno B7-1/metabolismo , Antígeno B7-2/genética , Antígeno B7-2/metabolismo , Células de la Médula Ósea/clasificación , Células de la Médula Ósea/citología , Células de la Médula Ósea/inmunología , Antígenos CD28/genética , Supervivencia Celular , Femenino , Inmunidad Humoral , Interleucina-6/biosíntesis , Masculino , Ratones , Ratones de la Cepa 129 , Ratones Endogámicos C57BL , Ratones Noqueados , Modelos Inmunológicos , Células Plasmáticas/clasificación , Células Plasmáticas/citología , Bazo/citología , Bazo/inmunología , Factores de TiempoRESUMEN
Conventional measurement of antibody responses to vaccines largely relies on serum antibodies, which are primarily produced by bone marrow plasma cells and may not represent the entire vaccine-induced B cell repertoire, including important functional components such as those targeted to mucosal sites. After immunization or infection, activated B cells differentiate into plasmablasts in local lymphoid organs, then traffic through circulation to the target sites where they further develop into plasma cells. On day 7 after influenza vaccination, a burst of plasmablasts, highly enriched for vaccine-specific antibody secreting cells, appears in the peripheral blood. This provides a unique window to the overall B cell response to the vaccine, without interference of pre-existing cross-reactive serum antibody. In this study we isolated B cells from volunteers on day 7 after immunization with the inactivated influenza vaccine and cultured them ex vivo to collect plasmablast-derived polyclonal antibodies (PPAb). The PPAb contained secreted IgG and IgA, which was approximately 0.2ng per antibody secreting cell. Influenza-specific IgG and IgA binding activity was detected in PPAb at dilutions up to 10(5) by ELISA. The ratio of the titers of influenza-specific IgA to IgG by ELISA was 4-fold higher in PPAb than in day 28 post-vaccination sera, suggesting that vaccine-induced IgA is enriched in PPAb compared to sera. Functional activity was also detected in PPAb as determined by microneutralization and hemagglutination inhibition assays. In addition to bulk B cell cultures, we also cultured plasmablast subsets sorted by cell surface markers to generate PPAb. These results suggest that PPAb better reflects the mucosal IgA response than serum samples. Since PPAb are exclusively produced by recently activated B cells, it allows assessing vaccine-induced antibody response without interference from pre-existing cross-reactive serum antibodies and permits an assessment of antibody avidity based on antigen specific binding and antibody quantity. Therefore this assay is particularly useful for studying vaccine/infection-induced antibodies against antigens that might have previously circulated, such as antibody responses to rotavirus, dengue or influenza viruses in which cross-reactive antibodies against different virus serotypes/subtypes play a critical role in immunity and/or pathogenesis.
Asunto(s)
Anticuerpos Antivirales/biosíntesis , Vacunas contra la Influenza/administración & dosificación , Células Plasmáticas/inmunología , Adolescente , Adulto , Anticuerpos Antivirales/sangre , Especificidad de Anticuerpos , Diferenciación Celular , Separación Celular , Células Cultivadas , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Pruebas de Inhibición de Hemaglutinación , Humanos , Inmunidad Mucosa , Inmunoglobulina A/biosíntesis , Inmunoglobulina A/sangre , Inmunoglobulina G/biosíntesis , Inmunoglobulina G/sangre , Técnicas In Vitro , Pruebas de Neutralización , Orthomyxoviridae/inmunología , Células Plasmáticas/clasificación , Células Plasmáticas/citología , Factores de Tiempo , Vacunación , Vacunas de Productos Inactivados/administración & dosificación , Adulto JovenRESUMEN
Although integration of laboratory data and radiological findings are required, quantity and qualitative evaluation of plasma cells in bone marrow is still useful for the diagnosis of multiple myeloma. However, there are considerable variations in morphology of myeloma and proliferation pattern among the individual patients. Therefore, it is important to recognize cytopathological features of myeloma cells for better understanding of patients' status. For example, the finding of immature plasmablastic myeloma cells predicts the poor prognosis of the patients. In the present review, we described the cytopathological features, which is useful for diagnosis and estimation of the patients' prognosis and discrimination from reactive non-tumorous plasma cells.
Asunto(s)
Células de la Médula Ósea/citología , Células de la Médula Ósea/patología , Mieloma Múltiple/diagnóstico , Mieloma Múltiple/patología , Células Plasmáticas/citología , Células Plasmáticas/patología , Núcleo Celular , Proliferación Celular , Citoplasma , Humanos , Células Plasmáticas/clasificación , PronósticoRESUMEN
Increases in free light chain (FLC) production are associated with disease progression in multiple myeloma (MM). Using a double immunofluorescence staining method to produce a differential count of plasma cells in bone marrow, single populations were demonstrated, containing intact monoclonal immunoglobulins (M-Igs) in 74% and FLCs only in 8% of cases. However, 18% contained a mixture of both cell populations. Progression from cells making intact M-Ig to cells restricted to FLC only production occurred in individual cases during the course of their disease. The presence of FLC only cells was associated with shortened survival.