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1.
Journal of Peking University(Health Sciences) ; (6): 975-981, 2023.
Artículo en Chino | WPRIM | ID: wpr-1010156

RESUMEN

OBJECTIVE@#To investigate the regulatory effect of interferon-α (IFN-α) on the apoptosis and killing function of CD56dimCD57+ natural killer (NK) cells in systemic lupus erythematosus (SLE) patients, and to explore the specific mechanism.@*METHODS@#A total of sixty-four newly treated SLE patients and sixteen healthy controls (HC) enrolled in the Second Hospital of Dalian Medical University were selected as the research subjects. And the gene expression levels of molecules related to NK cell-killing function were detected by real-time quantitative polymerase chain reaction. CD56dimCD57+ NK cells were co-cultured with the K562 cells, and the apoptotic K562 cells were labeled with Annexin-Ⅴ and 7-amino-actinomycin D. Peripheral blood mononuclear cells were treated with 20, 40, and 80 μmol/L hydrogen peroxide (H2O2), and treated without H2O2 as control, the expression level of perforin (PRF) was detected by flow cytometry. The concentration of IFN-α in serum was determined by enzyme linked immunosorbent assay. The expression levels of IFN-α receptors (IFNAR) on the surface of CD56dimCD57+ NK cells were detected by flow cytometry, and were represented by mean fluorescence intensity (MFI). CD56dimCD57+ NK cells were treated with 1 000 U/mL IFN-α for 24, 48 and 72 h, and no IFN-α treatment was used as the control, the apoptosis and the expression levels of mitochondrial reactive oxygen species (mtROS) were measured by flow cytometry and represented by MFI.@*RESULTS@#Compared with HC(n=3), the expression levels of PRF1 gene in peripheral blood NK cells of the SLE patients (n=3) were decreased (1.24±0.41 vs. 0.57±0.12, P=0.05). Compared with HC(n=5), the ability of peripheral blood CD56dimCD57+ NK cells in the SLE patients (n=5) to kill K562 cells was significantly decreased (58.61%±10.60% vs. 36.74%±6.27%, P < 0.01). Compared with the control (n=5, 97.51%±1.67%), different concentrations of H2O2 treatment significantly down-regulated the PRF expression levels of CD56dimCD57+ NK cells in a dose-dependent manner, the 20 μmol/L H2O2 PRF was 83.23%±8.48% (n=5, P < 0.05), the 40 μmol/L H2O2 PRF was 79.53%±8.56% (n=5, P < 0.01), the 80 μmol/L H2O2 PRF was 76.67%±7.16% (n=5, P < 0.01). Compared to HC (n=16), the serum IFN-α levels were significantly increased in the SLE patients (n=45) with moderate to high systemic lupus erythematosus disease activity index (SLEDAI≥10) [(55.07±50.36) ng/L vs. (328.2±276.3) ng/L, P < 0.001]. Meanwhile, compared with HC (n=6), IFNAR1 expression in peripheral blood CD56dimCD57+ NK cells of the SLE patients (n=6) were increased (MFI: 292.7±91.9 vs. 483.2±160.3, P < 0.05), and compared with HC (n=6), IFNAR2 expression in peripheral blood CD56dimCD57+ NK cells of the SLE patients (n=7) were increased (MFI: 643.5±113.7 vs. 919.0±246.9, P < 0.05). Compared with control (n=6), the stimulation of IFN-α (n=6) significantly promoted the apoptosis of CD56dimCD57+ NK cells (20.48%±7.01% vs. 37.82%±5.84%, P < 0.05). In addition, compared with the control (n=4, MFI: 1 049±174.5), stimulation of CD56dimCD57+ NK cells with IFN-α at different times significantly promoted the production of mtROS in a time-dependent manner, 48 h MFI was 3 437±1 472 (n=4, P < 0.05), 72 h MFI was 6 495±1 089 (n=4, P < 0.000 1), but there was no significant difference at 24 h of stimulation.@*CONCLUSION@#High serum IFN-α level in SLE patients may induce apoptosis by promoting mtROS production and inhibit perforin expression, which can down-regulate CD56dimCD57+ NK killing function.


Asunto(s)
Humanos , Interferón-alfa/metabolismo , Perforina/metabolismo , Leucocitos Mononucleares/metabolismo , Peróxido de Hidrógeno/metabolismo , Interferón gamma/metabolismo , Antígeno CD56/metabolismo , Células Asesinas Naturales/metabolismo , Lupus Eritematoso Sistémico
2.
Journal of Experimental Hematology ; (6): 553-561, 2023.
Artículo en Chino | WPRIM | ID: wpr-982094

RESUMEN

OBJECTIVE@#To explore the similarities and variations of biological phenotype and cytotoxicity of human umbilical cord blood natural killer cells (hUC- NK) after human umbilical cord blood-derived mononuclear cells (hUC-MNC) activated and expanded by two in vitro high-efficient strategies.@*METHODS@#Umbilical cord blood mononuclear cells (MNC) from healthy donor were enriched by Ficoll-based density gradient centrifugation. Then, the phenotype, subpopulations, cell viability and cytotoxicity of NK cells derived from Miltenyi medium (denoted as M-NK) and X-VIVO 15 (denoted as X-NK) were compared using a "3IL" strategy.@*RESULTS@#After a 14-day's culture, the contents of CD3-CD56+ NK cells were elevated from 4.25%±0.04% (d 0) to 71%±0.18% (M-NK) and 75.2%±1.1% (X-NK) respectively. Compared with X-NK group, the proportion of CD3+CD4+ T cells and CD3+CD56+ NKT cells in M-NK group decreased significantly. The percentages of CD16+, NKG2D+, NKp44+, CD25+ NK cells in X-NK group was higher than those in the M-NK group, while the total number of expanded NK cells in X-NK group was half of that in M-NK group. There were no significant differences between X-NK and M-NK groups in cell proliferation and cell cycle, except for the lower percentage of Annexin V+ apoptotic cells in M-NK group. Compared with X-NK group, the proportion of CD107a+ NK cells in M-NK group were higher under the same effector-target ratio (E∶T) (P<0.05).@*CONCLUSION@#The two strategies were adequate for high-efficient generation of NK cells with high level of activation in vitro, however, there are differences in biological phenotypes and tumor cytotoxicity.


Asunto(s)
Humanos , Sangre Fetal , Células Asesinas Naturales , Linfocitos T , Leucocitos Mononucleares/metabolismo , Proliferación Celular , Antígeno CD56/metabolismo
3.
Hematol., Transfus. Cell Ther. (Impr.) ; 44(2): 143-150, Apr.-June 2022. tab, graf
Artículo en Inglés | LILACS | ID: biblio-1385039

RESUMEN

Abstract Introduction Flow cytometric immunophenotyping (FCI) plays a major role in diagnosing hematologic malignancies. In patients diagnosed with precursor B-lineage acute lymphoblastic leukemia (B-ALL), expression of certain non-lineage/cross lineage antigens is of prognostic and cytogenetic relevance. There is a paucity of studies that have comprehensively analyzed the clinical and laboratory profiles of B-ALL patients showing aberrant T/natural killer (NK) cell antigen expression. Materials and methods This is a prospective study where 152 consecutive B-ALL patients were analyzed for aberrant expression of T/NK cell antigens (CD1a, CD5, CD4, CD7, CD8 and CD56) by FCI. The clinical and laboratory profile of these T/NK-cell antigen-expressing B-ALL patients was statistically analyzed against conventional B-ALL patients. Results In our B-ALL cohort, CD5, CD7 and CD56 expression were observed in one, six and nine patients, respectively. CD56-expressing B-ALL patients were predominantly children (89%) and presented as standard clinical risk (p = 0.010) disease with frequent ETV6-RUNX1 fusion (p = 0.021) positivity. On the contrary, CD7-expressing B-ALL patients were adolescent-young adult/adult-age skewed (83%) and had an adverse cytogenetic profile (p = 0.001), especially for the frequent presence of BCR-ABL1 fusion (p = 0.004) and KMT2A rearrangement (p = 0.045). CD7-expressing B-ALL patients had inferior event-free survival (p = 0.040) than their CD56-expressing counterparts, but there was no significant difference in the overall survival (p = 0.317). Conclusion In comparison to conventional B-ALL patients, there are significant differences in the age, cytogenetic profile and event-free survival of T/NK-cell antigen-expressing B-ALL patients.


Asunto(s)
Humanos , Masculino , Femenino , Lactante , Preescolar , Niño , Adolescente , Adulto , Persona de Mediana Edad , Adulto Joven , Leucemia-Linfoma Linfoblástico de Células Precursoras B , Citometría de Flujo , Inmunofenotipificación , Antígenos CD7 , Antígeno CD56
4.
Journal of Experimental Hematology ; (6): 49-55, 2022.
Artículo en Chino | WPRIM | ID: wpr-928669

RESUMEN

OBJECTIVE@#To explore the expression characteristics of antigens and functional markers of natural killer (NK) cells in patients with acute myeloid leukemia (AML).@*METHODS@#Multi-parameter flow cytometry was used to detect NK cell surface markers and their functional indicators in 56 newly diagnosed AML patients and 24 healthy controls, including activating receptors NKG2D, NKP46, DNAM-1, and killing indicators granzyme B, perforin.@*RESULTS@#Referring to the WHO hematopoiesis and lymph tissue tumor classification criteria, 56 cases were roughly divided into three types: AML M1, M2, and M4/M5. However, there was no differences about NK cells among the three types, so it was no longer subdivided. NK cells were divided into two groups: CD3-CD56hiCD16- (CD56hiNK) and CD3-CD56dimCD16+ (CD56dimNK). Compared with CD56dimNK cell population, except for NKP46, the positive expression levels of NKG2D and other receptors of CD56hiNK cells in AML patients decreased (P<0.001). Compared with healthy controls, the proportion of CD56hiNK cells in AML patients increased, while the number and proportion of NK cells and proportion of CD56dimNK cells significantly decreased (P<0.05). The proportion of perforin in CD56hiNK cells significantly increased (P<0.05). The expression of DNAM-1 in CD56hiNK cells, NKG2D, DNAM-1, and perforin in CD56dimNK cells decreased significantly (P<0.05). There was no statistically significant difference in expression of other functional indexes in AML patients compared with corresponding indexes of healthy controls. In addition, the proportion of CD56hiNK cells was positively correlated with the expression of CD34+ in AML (r=0.303).@*CONCLUSION@#Compared with CD56dimNK, the ratio of CD56hiNK and the expression of functional markers in AML patients are lower. Compared with healthy controls, the number and expression ratio of NK cells in AML patients decrease and the expression of functional markers is abnormal, indicating that its function is impaired.


Asunto(s)
Humanos , Antígeno CD56 , Citometría de Flujo , Células Asesinas Naturales , Leucemia Mieloide Aguda
5.
Journal of Experimental Hematology ; (6): 827-831, 2021.
Artículo en Chino | WPRIM | ID: wpr-880154

RESUMEN

OBJECTIVE@#To investigate the significance of CD27 and CD56 in the prognosis of multiple myeloma (MM) patients, and to establish a simple and convenient prognostic risk score.@*METHODS@#One hundred and eleven newly diagnosed MM patients treated by bortezomib in Shengjing hospital from January 1, 2013 to January 1, 2019 were selected, and the relationship between clinical characteristics and survival time of patients was analyzed.@*RESULTS@#The overall survival (OS) of patients in CD27@*CONCLUSION@#Among patients with MM treated by bortezomib, CD27


Asunto(s)
Humanos , Bortezomib , Antígeno CD56 , Trasplante de Células Madre Hematopoyéticas , Mieloma Múltiple , Pronóstico , Estudios Retrospectivos
6.
Journal of Experimental Hematology ; (6): 553-556, 2021.
Artículo en Chino | WPRIM | ID: wpr-880112

RESUMEN

OBJECTIVE@#To investigate the expression of CD56 in multiple myeloma (MM) cells and its relationship between extramedullary disease and extramedullary relapse.@*METHODS@#Clinical data of 99 patients with MM treated in our hospital from January 2015 to December 2019 was retrospectively analyzed. The patients were divided into positive group and negative group according to the expression of CD56. The relationship between CD56 and multiple myeloma extramedullary disease, extramedullary relapse was analyzed.@*RESULTS@#Among 99 newly diagnosed patients with MM, the positive rate of CD56 was 65%, and the incidence of extramedullary disease of patients in the CD56 positive group was lower than that in the CD56 negative group (17.19% vs 48.57%) (P<0.01). Meanwhile, the incidence of extramedullary relapse of patients in the CD56 positive group was lower than that in the CD56 negative group (1.56% vs 34.29%) (P<0.01).@*CONCLUSION@#CD56 is highly expressed in MM, and its low expression is associated with the occurrence of extramedullary disease and extramedullary relapse, which suggests that CD56 may be an important indicator for predicting the occurrence of extramedullary disease and extramedullary relapse.


Asunto(s)
Humanos , Antígeno CD56 , Mieloma Múltiple , Recurrencia Local de Neoplasia , Estudios Retrospectivos
7.
Gac. méd. Méx ; 156(3): 188-194, may.-jun. 2020. tab, graf
Artículo en Inglés, Español | LILACS | ID: biblio-1249893

RESUMEN

Resumen Introducción: Después de un trasplante de células progenitoras hematopoyéticas (TCPH), la reconstitución de las células natural killer (NK) es la principal barrera contra las infecciones virales. Objetivo: Determinar que el conocimiento sobre la cinética de la reconstitución de las células NK posterior al TCPH contribuye a un eficiente monitoreo del trasplante, lo que incrementa la posibilidad de éxito de este. Método: Se incluyeron 21 pacientes sometidos a TCPH, así como un grupo control de individuos clínicamente sanos. En diferentes momentos después del trasplante (intervalo de 21 a 670 días), mediante citometría de flujo se cuantificaron las células NK CD3− CD16+ CD56+ en muestras de sangre periférica. Resultados: La recuperación de las células NK ocurre entre los tres y seis meses y entre los 10 y 12 meses postrasplante; su número fue significativamente menor (en comparación con el grupo control) en el tiempo restante del monitoreo. Conclusiones: El primer periodo de recuperación de las células NK ocurre entre los tres y seis meses posteriores al trasplante. La reconstitución es transitoria y el número de células NK varía en los primeros años.


Abstract Introduction: After hematopoietic stem cell transplantation (HSCT), natural killer (NK) cells reconstitution is the main barrier against viral infections. Objective: To determine that the knowledge on the kinetics of NK cell reconstitution after HSCT contributes to transplant efficient monitoring, which increases the possibility of its success. Method: Twenty-one patients undergoing HSCT were included, as well as a control group of clinically healthy individuals. At different time points after transplantation (range of 21 to 670 days), CD3- CD16+ CD56+ NK cells were quantified by flow cytometry in peripheral blood samples. Results: NK cell recovery occurs at three to six months and 10 to 12 months post-transplantation; their number was significantly lower (in comparison with the control group) in the rest of the monitoring time. Conclusions: The first period of NK cell recovery occurs between three and six months after transplantation. Reconstitution is transient and the number of NK cells varies in the first years.


Asunto(s)
Humanos , Masculino , Femenino , Lactante , Preescolar , Niño , Adolescente , Células Asesinas Naturales/citología , Trasplante de Células Madre Hematopoyéticas/métodos , Factores de Tiempo , Estudios Prospectivos , Receptores de IgG , Complejo CD3 , Antígeno CD56 , Proteínas Ligadas a GPI , Citometría de Flujo
8.
Int. j. morphol ; 38(2): 247-251, abr. 2020. tab, graf
Artículo en Inglés | LILACS | ID: biblio-1056430

RESUMEN

Nine tumor and various potential biomarkers were measured and combined the information to diagnose disease, all patients accepted fiber bronchoscopy brush liquid based cytologyand histopathology examination in order to reliably detect lung cancer. The samples from 314 Chinese lung cancer patients were obtained and CK5/6, P63, P40, CK7, TTF-1, NapsinA CD56, Syn and CgA were measured with the immunohistochemical SP method and analyzed correlation of the expression of these markers with pathological and clinical features of squamous cell carcinoma, adenocarcinoma, and small cell lung carcinoma. Squamous cell carcinoma, adenocarcinoma and small cell carcinoma were 61 cases, 114 cases and 139 cases,CK5/6 and P63 expression were more frequent in squamous cell carcinoma, with sensitivity and specificity of 77.05 % and 96.44 %, 83.61 % and 88.93 %,and compared with adenocarcinoma and small cell carcinoma difference was statistically significant (P<0.05), The incidences of a positive P40 expression were 100 % in squamous cell carcinoma, with specificity of 98.81 %.CK7, TTF-1 and NapsinA expression were more frequent in adenocarcinoma, with sensitivity and specificity of 85.09 % and 78.69 %, 79.82 % and 93.44 %, 56.14 % and 95.08 %, and compared with squamous cell carcinoma and small cell carcinoma difference was statistically significant (P<0.05). TTF-1, Syn, CgA and CD56 expression were more frequent in adenocarcinoma, with sensitivity and specificity of 86.33 % and 93.44 %, 89.21 % and 98.36 %, 74.10 % and 100 %, 96.40 % and 96.72 %. The combined detection of CK5/6, P63 and P40 were more useful and specific in differentiating squamous cell carcinoma. CK7, TTF-1 and NapsinA were more useful and specific in differentiating lung adenocarcinoma. The impaired CD56, TTF-1, Syn and CgA reflects the progression of small cell lung cancer.


Se midieron tumores y utilizaron nueve biomarcadores potenciales y se analizó la información para diagnosticar la enfermedad. A todos los pacientes se les realizó citología en líquido con broncoscopía de fibra y examen histopatológico para detectar de manera confiable el cáncer pulmonar. Se obtuvieron muestras de 314 pacientes chinos con cáncer de pulmón y CK5 / 6, P63, P40, CK7, TTF-1, Napsina A, CD56, Syn y CgA se midieron a través de histoquímica SP y analizaron la correlación de la expresión de estos marcadores con características patológicas y clínicas de carcinoma de células escamosas, adenocarcinoma y carcinoma de células pequeñas en el cáncer de pulmón. El carcinoma de células escamosas, el adenocarcinoma y el carcinoma de células pequeñas fueron 61 casos, 114 casos y 139 casos, respectivamente, la expresión de CK5 / 6 y P63 fueron más frecuentes en el carcinoma de células escamosas, con una sensibilidad y especificidad del 77,05 % y 96,44 %, 83,61 % y 88,93 %, y en comparación con el adenocarcinoma y el carcinoma de células pequeñas, la diferencia fue estadísticamente significativa (P <0,05). La incidencia de ap la expresión positiva P40 fue del 100 % en el carcinoma de células escamosas, con una especificidad del 98,81 %. La expresión de CK7, TTF-1 y NapsinA fueron más frecuentes en el adenocarcinoma, con una sensibilidad y especificidad del 85,09 % y 78,69 %, 79,82 % y 93,44 %, 56,14 % y 95,08 %, y en comparación con el carcinoma de células escamosas y la diferencia de carcinoma de células pequeñas fue estadísticamente significativa (P <0,05) .TTF-1, Syn, CgA y la expresión de CD56 fueron más frecuentes en adenocarcinoma, con sensibilidad y especificidad de 86.33 % y 93.44 %, 89.21 % y 98.36 %, 74.10 % y 100 %, 96.40 % y 96.72 %. La detección combinada de CK5 / 6, P63 y P40 fue más útil y específica en la diferenciación del carcinoma de células escamosas. CK7, TTF-1 y NapsinA fueron más útiles y específicos para diferenciar el adenocarcinoma de pulmón. El deterioro de CD56, TTF-1, Syn y CgA refleja la progresión del cáncer de pulmón de células pequeñas.


Asunto(s)
Humanos , Carcinoma/metabolismo , Carcinoma/patología , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Fragmentos de Péptidos/metabolismo , Factores de Transcripción/metabolismo , Inmunohistoquímica , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patología , Adenocarcinoma/metabolismo , Adenocarcinoma/patología , Biomarcadores de Tumor/metabolismo , Ácido Aspártico Endopeptidasas/metabolismo , Sensibilidad y Especificidad , Carcinoma de Células Pequeñas/metabolismo , Carcinoma de Células Pequeñas/patología , Antígeno CD56/metabolismo , Proteínas Supresoras de Tumor/metabolismo , Queratinas Tipo II/metabolismo , Queratina-7/metabolismo , Factor Nuclear Tiroideo 1/metabolismo
9.
Journal of Experimental Hematology ; (6): 1899-1903, 2020.
Artículo en Chino | WPRIM | ID: wpr-879990

RESUMEN

OBJECTIVE@#To investigate the expression of costimulatory molecule Tim3 and its subsets on NK cells in patients with acute myeloid leukemia.@*METHODS@#30 patients with acute myeloid leukemia treated in our hospital from August 2016 to August 2018 were randomly selected as the AML group, and 30 healthy persons in our hospital during the same period were randomly selected as the control group. The expression levels of CD56@*RESULTS@#The expression levels of CD56@*CONCLUSION@#The expression of costimulatory molecule Tim3 in NK cells and its subsets of patients with acute myeloid leukemia is down-regulated.


Asunto(s)
Humanos , Antígeno CD56 , Citometría de Flujo , Receptor 2 Celular del Virus de la Hepatitis A , Células Asesinas Naturales , Leucemia Mieloide Aguda , Pacientes
10.
Braz. dent. j ; 29(4): 400-404, July-Aug. 2018. graf
Artículo en Inglés | LILACS | ID: biblio-974165

RESUMEN

Abstract Melanotic neuroectodermal tumor of infancy (MNTI) is a rare neoplasm that affects mainly children under 1 year of age. A 4-month-old boy was referred for evaluation of a lesion with 1 month of evolution. Intra-oral examination detected a firm upon palpation submucosal nodular mass, measuring 1.5 cm in diameter, affecting the anterior maxillary alveolar ridge and covered by a slightly blue mucosa with evident telangiectasia. The patient underwent an incisional biopsy and histological and immunohistochemical analyses revealed nests of AE1/AE3 positive epithelioid cells with abundant melanin pigmentation. Other cell types, resembling neuroblasts, were also present and positive for CD56, synaptophysin and enolase. The diagnosis of MNTI was established and the patient was referred for treatment. Conservative surgical resection was performed along with 3 adjacent teeth under general anesthesia. The patient is in follow-up for 1,5 year without recurrence. Conservative surgical management of MNTI may be an alternative to maxillectomy, contributing to the patient´s quality of life.


Resumo Tumor neuroectodérmico melanótico da infância (TNMI) é um neoplasma raro que afeta principalmente crianças com idade abaixo de 1 ano. Um menino de 4 meses foi referenciado para avaliação de uma lesão com 1 mês de evolução. O exame intra-oral detectou uma massa nodular submucosa firme à palpação, medindo 1,5 cm de diâmetro, afetando rebordo alveolar anterior da maxila e recoberta por mucosa de coloração levemente azulada com telangiectasia evidente. O paciente foi submetido à biopsia incisional e as análises histológica e imunohistoquímica revelaram ninhos compostos por células com abundante pigmento de melanina, positivas para AE1/AE3. Outro tipo celular, semelhante à neuroblastos, também estava presente e foram positivas para CD56, sinaptofisina e enolase. O diagnóstico de TNMI foi estabelecido e o paciente encaminhado para tratamento. Ressecção cirúrgica conservadora sob anestesia geral ao longo de 3 dentes adjacentes foi realizada. O paciente está em acompanhamento há 1 ano e meio sem sinais de recorrência. O tratamento cirúrgico conservador do TNMI pode ser uma alternativa à maxilectomia, contribuindo para a qualidade de vida do paciente.


Asunto(s)
Humanos , Masculino , Lactante , Tumor Neuroectodérmico Melanótico/diagnóstico , Biopsia , Inmunohistoquímica , Tumor Neuroectodérmico Melanótico/cirugía , Tumor Neuroectodérmico Melanótico/metabolismo , Tumor Neuroectodérmico Melanótico/patología , Antígeno CD56/metabolismo , Melaninas/metabolismo
11.
Journal of Experimental Hematology ; (6): 547-551, 2018.
Artículo en Chino | WPRIM | ID: wpr-690952

RESUMEN

<p><b>OBJECTIVE</b>To develop an easy method to amplify natural killer (NK) cells by using mononuclear cells in vitro, so as to lay the basis for NK cell therapy.</p><p><b>METHODS</b>Umbilical cord blood from 3 healthy full-term pregnant women was collected, and the peripheral blood mononuclear cells (PBMNC) were harvested by density gradient centrifugation. Each sample of PBMNC was divided into 3 groups: CD16mAb, CD3 mAb and CD16mAb+CD3mAb- groups. The culture flasks were pre-coated with CD16, CD3 or CD3 plus CD16 mAb. The PBMNCs were cultured in serum-free media containing autologous plasma, recombinant human IL-2, IL-15 and IL-21 for 14 days under the same conditions. The total viable cell count was calculated. Flow cytometry was used to determine the ratio of CD56CD3 cells, MTT assay was used to measure the killing rate of NK cells under different effector/target ratio, by using the K562 cells as the target cells.</p><p><b>RESULTS</b>After 14 days of culture, the total cell numbers of CD16mAb, CD3mAb and CD16mAb +CD3mAb groups increased by 45.71±5.54, 87.41±19.77 and 4.88±51.84 times, respectively, and those of CD3mAb group were significantly higher than the other 2 groups (P<0.05). The ratio of CD56CD3 cells before culture was 0.1663±0.0201, which was 0.8167±0.0500, 0.8077±0.0589 and 0.8077±0.0273 after incubation with CD16mAb, CD3mAb and CD16mAb +CD3mAb for 14 days, respectively (P>0.05). MTT test showed that the killing efficiencies were not significantly different among the 3 groups when the effector/target ratios were 1:1, 5:1 and 10:1 (P>0.05).</p><p><b>CONCLUSION</b>By incubation with anti-CD3 monoclonal antibody, IL-2, IL-15 and IL-21, the highly purified NK cells can be obtained from mononucleated cells, thus providing a simple method for NK cell therapy.</p>


Asunto(s)
Femenino , Humanos , Embarazo , Complejo CD3 , Antígeno CD56 , Técnicas de Cultivo de Célula , Células Cultivadas , Células Asesinas Naturales , Leucocitos Mononucleares
12.
Journal of Experimental Hematology ; (6): 552-556, 2018.
Artículo en Chino | WPRIM | ID: wpr-690951

RESUMEN

<p><b>OBJECTIVE</b>To establish a novel method for ex vivo expansion of natural killer cells from human umbilical blood, so as to provide the basis for NK cell therapy.</p><p><b>METHODS</b>Mononucleated cells from human umbilical blood were harvested and suspended in a serum-free medium containing 5% autologous plasma, recombinant human IL-15 (50 ng/ml) and hydrocortisone sodium succinate (5×10 mol/L) at a concentration of 1.5×10/ml, then the cells were seeded into flasks pre-coated with heparin sodium (100 U/cm) or/and anti-human CD16 antibody (1 µg/cm). After culture for 2 weeks, the cells were harvested and counted. Ratios of CD3/CD56 of the cells were determined by flow cytometry. MTT test was performed to assess the cytotoxicity against K562 cells with graded ratios of effector/target cells.</p><p><b>RESULTS</b>In contrast to the cells in flasks without pre-coating, the attached colonies appeared predominantly within 1 week of culture from heparin- and antibody-coated groups. The cell numbers from the pre-coated groups were significantly higher than that of uncoated one after culture for 2 weeks. Furthermore, the ratios of CD3/CD56 cells were much higher in pre-coated groups, and that of the cells from flasks pre-coated with heparin and antibody were the highest (all the P values <0.01). MTT test showed that the cytotoxic activity of the cells stimulated by precoating were much more potent than that of the cells without the stimulation.</p><p><b>CONCLUSION</b>Advantageous expansion of NK cells can be achieved by precoating with heparin and anti-CD16 antibody, and also by supplement with IL-15 and hydrocortisone into the media, so the umbilical NK cells with high purity and potent cytotoxicity can be obtained.</p>


Asunto(s)
Humanos , Antígeno CD56 , Células Cultivadas , Citotoxicidad Inmunológica , Sangre Fetal , Heparina , Células Asesinas Naturales
13.
Journal of Experimental Hematology ; (6): 727-732, 2018.
Artículo en Chino | WPRIM | ID: wpr-689585

RESUMEN

<p><b>OBJECTIVE</b>To investigate the clinical significance of RUNX1-RUNX1T1 expression level in bone marrow of patients with acute non-M3 myeloid leukemia (AML non-M3), and to understand the biological characteristics of RUNX1-RUNX1T1 positive AML expressing lymphoid antigens CD19, CD56 and its effect on the initially induced remission rate and prognosis.</p><p><b>METHODS</b>The expression level of RUNX1-RUNX1T1 in bone marrow of 200 patients with newly diagnosed AML (non-M3) was detected by real-time fluorescent Q-PCR, the expression level of lymphoid antigens was detected by flow cytometry, and the relationship of the initially induced remission rate (CR1) with the overall survival (OS) rate was analyzed, the CR1 and OS differences also were analyzed between CD56 and CD56 patients as well as CD19 and CD17 patients in RUNX1-RUNX1T1 positive patients with AML.</p><p><b>RESULTS</b>The CD56 patients at the initial diagnosis had lower CR1(P<0.05) in RUNX1-RUNX1T1 positive AML patients, the CR1 of CD19 patients was higher than that in CD19 patients at the initial diagnosis (P<0.05). The OS of CD56 patients was significantly high in comparison with CD56 patients (P<0.05), while the OS between CD19 patients and CD19 patients was not significantly different.</p><p><b>CONCLUSION</b>The bone marrow CD56 in RUNX1-RUNX1T1 positive AML patients suggests poor prognosis. The CD19 only correlates with CR1, but does not with OS.</p>


Asunto(s)
Humanos , Antígenos CD19 , Antígeno CD56 , Subunidad alfa 2 del Factor de Unión al Sitio Principal , Leucemia Mieloide Aguda , Mutación , Pronóstico , Proteína 1 Compañera de Translocación de RUNX1
14.
Egyptian Journal of Pediatric Allergy and Immunology [The]. 2017; 15 (2): 69-69
en Inglés | IMEMR | ID: emr-189228
15.
Appl. cancer res ; 37: 1-6, 2017. ilus
Artículo en Inglés | LILACS, Inca | ID: biblio-911493

RESUMEN

Background: Atypical teratoid/rhabdoid tumors (AT/RT) are aggressive embryonal tumors of the central nervous system. They are largely characterized by inactivating mutations of the SMARCB1 tumor suppressor gene. AT/RT patients have a very poor prognosis and no standard therapeutic protocol has been defined yet. Recently, multimodal therapy with multiple drug combinations has slightly improved the overall survival, however drug toxicity remains high. In this scenario, a better understanding of the pathophysiology of the disease is needed. Methods: We evaluated the gene expression profile of AT/RT samples to find new genetic factors contributing to the pathophysiology of the disease. We found target genes significantly differentially expressed between AT/RT and medulloblastoma (MB), the most common embryonal brain tumor. The mRNA expression was validated by quantitative real-time PCR and, at the protein level, expression was validated by immunohistochemistry in an independent set of tumors. Results: The Neural cell adhesion molecule 1 (NCAM1) gene was found to be consistently downregulated in AT/RT samples when compared to MB and normal brain tissue. Immunohistochemistry showed that the expression of NCAM1 in AT/RT was significantly lower than that of MB. Conclusion: NCAM1 is an important molecule involved in neuron-to-neuron and neuron-to-muscle adhesion during development. Downregulation of NCAM1 has been implicated in several human cancers suggesting that it might have a tumor repressor role. In this study we found a significantly reduced expression of NCAM1 in AT/RT when compared to MB and we suggest that this feature can be used as a diagnostic marker, along with demonstration of SMARCB1 (INI1) or SMARCA4 (BRG1) inactivation. The roles of NCAM1 in the pathophysiology of AT/RT are still to be determined (AU)


Asunto(s)
Humanos , Teratoma/diagnóstico , Inmunoglobulinas , Biomarcadores de Tumor , Tumor Rabdoide/diagnóstico , Antígeno CD56
16.
Chinese Journal of Pathology ; (12): 78-82, 2016.
Artículo en Chino | WPRIM | ID: wpr-278560

RESUMEN

<p><b>OBJECTIVE</b>To study the clinicopathologic features and significance of aberrant CD56 expression in diffuse large B-cell lymphoma (DLBCL).</p><p><b>METHODS</b>The clinical and pathologic profiles of 10 cases of DLBCL with aberrant expression of CD56 were investigated. Immunohistochemical staining, in-situ hybridization for Epstein-Barr virus encoded RNA and gene rearrangement for IgH and Igκ were carried out.</p><p><b>RESULTS</b>There were 6 male and 4 female patients. The medium age of patients was 46 years. All of them presented with extranodal lymphoma involvement, with gastrointestinal tract being the commonest site (5/10). Histologic examination showed that most of the atypical lymphoid cells were centroblast-like and demonstrated a diffuse growth pattern. Apoptosis and necrosis were identified in some cases. Immunohistochemical study showed that the tumor cells were positive for CD20 or CD79α and aberrantly expressed CD56. Five cases had the GCB phenotype while the remaining cases had the non-GCB phenotype, according to Hans classification. Bcl-6 was positive in most cases (9/10). All cases showed a high proliferation index by Ki-67. The tumor cells were negative for CD3ε, CD138 and granzyme B. In-situ hybridization for Epstein-Barr virus encoded RNA was performed in 7 cases and none of them showed positive signals. IgH gene rearranged bands were detected in 4 cases (4/6) and Igκ was detected in 3 cases (3/6). Follow-up data were available in 8 patients. Two patients died of disease progression within 5 to 13 months after diagnosis and the other 6 patients were alive 8 to 60 months after therapy.</p><p><b>CONCLUSIONS</b>DLBCL with aberrant expression of CD56 is rare. Most of them present with extranodal involvement, show high frequency of bcl-6 expression and high proliferation index. The patients often have good response to chemotherapy.</p>


Asunto(s)
Femenino , Humanos , Masculino , Persona de Mediana Edad , Antígenos CD20 , Metabolismo , Apoptosis , Antígeno CD56 , Metabolismo , Antígenos CD79 , Metabolismo , Progresión de la Enfermedad , Reordenamiento Génico , Granzimas , Metabolismo , Herpesvirus Humano 4 , Genética , Inmunofenotipificación , Hibridación in Situ , Linfoma de Células B Grandes Difuso , Genética , Metabolismo , Patología , Necrosis , Fenotipo , Proteínas Proto-Oncogénicas c-bcl-6 , Metabolismo , ARN Viral
18.
Chinese Journal of Pathology ; (12): 644-647, 2015.
Artículo en Chino | WPRIM | ID: wpr-358946

RESUMEN

<p><b>OBJECTIVE</b>To study the expression of squamous cell markers p63, p40 and CK5/6 in small cell carcinoma of lung (SCLC).</p><p><b>METHODS</b>Immunohistochemical study for squamous cell markers (p63, p40 and CK5/6), neuroendocrine markers (chromogranin A, synaptophysin and CD56) and TTF1 was carried out in 283 cases of SCLC. The diagnostic value of these markers was evaluated.</p><p><b>RESULTS</b>The expression rate of p63, p40 and CK5/6 were 20.7% (54/261), 7.9% (5/63) and 0.5% (1/221), respectively in the cases of SCLC studied. Amongst the squamous cell markers, CK5/6 had the lowest rate of positivity (P < 0.01). On the other hand, chromogranin A, synaptophysin and CD56 were positive in 61.8% (170/275), 85.5% (242/283) and 89.2% (248/278), respectively. The positivity rate for chromogranin A was lower than that for synaptophysin and CD56 (P < 0.01). TTF1 was expressed in 77.2% (217/281).</p><p><b>CONCLUSIONS</b>p63 and p40 are expressed in a subset of SCLC. In contrast, CK5/6 is rarely positive in SCLC. An immunohistochemical panel of CK5/6, synaptophysin and CD56 is recommended for differential diagnosis of SCLC.</p>


Asunto(s)
Humanos , Antígeno CD56 , Genética , Metabolismo , Cromogranina A , Genética , Metabolismo , Proteínas de Unión al ADN , Genética , Metabolismo , Diagnóstico Diferencial , Queratina-5 , Genética , Metabolismo , Queratina-6 , Genética , Metabolismo , Neoplasias Pulmonares , Genética , Metabolismo , Carcinoma Pulmonar de Células Pequeñas , Genética , Metabolismo , Sinaptofisina , Genética , Metabolismo , Factores de Transcripción , Genética , Metabolismo , Proteínas Supresoras de Tumor , Genética , Metabolismo
19.
Chinese Journal of Hematology ; (12): 676-681, 2015.
Artículo en Chino | WPRIM | ID: wpr-296176

RESUMEN

<p><b>OBJECTIVE</b>To explore the clinical features and survival of patients with CD56 expression in de- novo acute myeloid leukemia(AML)with t(8;21). .</p><p><b>METHODS</b>Clinical data of 82 de novo AML with t(8;21)who were newly diagnosed from Jan 2008 to Apr 2014 were analyzed retrospectively, 50 expressed CD56 and 32 not. Clinical characteristics and prognoses were compared between patients expressing and nonexpressing CD56.</p><p><b>RESULTS</b>There were no statistically significant differences in terms of age, gender, white blood cell count(WBC), percentage of bone marrow blasts, extramedullary infiltration rate, the early mortality or the presence of additional cytogenetic abnormalities between CD56 + and CD56- groups(P>0.05). The expressions of lymphatic antigens CD19 between CD56 + and CD56- groups showed significant difference (30.0% vs 53.1% , P=0.036). The complete remission and 3-year overall survival(OS)showed no significant differences between CD56+ and CD56-groups, while 3- year disease- free survival(DFS)showed significant differences(25.8% vs 46.9%, P=0.014). Multivariable analysis for DFS identified CD56 positivity as an independent predictor. DFS of who received allogeneic hematopoietic stem cell transplantation(HSCT)was better than those treated with intermediate- dose cytarabine/high dose cytarabine(IDAC)as postremission therapy.</p><p><b>CONCLUSION</b>The expression of CD56 in de-novo AML with t(8;21) appeared to be associated with poorer prognosis.</p>


Asunto(s)
Humanos , Médula Ósea , Antígeno CD56 , Aberraciones Cromosómicas , Cromosomas Humanos Par 21 , Cromosomas Humanos Par 8 , Citarabina , Supervivencia sin Enfermedad , Trasplante de Células Madre Hematopoyéticas , Leucemia Mieloide Aguda , Pronóstico , Inducción de Remisión , Estudios Retrospectivos , Análisis de Supervivencia
20.
Journal of Experimental Hematology ; (6): 1133-1137, 2015.
Artículo en Chino | WPRIM | ID: wpr-274080

RESUMEN

<p><b>OBJECTIVE</b>To study the amount change of peripheral blood NK cells in patients with hematologic malignancies and its significance.</p><p><b>METHODS</b>A total of 200 patients with hematologic malignancies in our hospital from June 2013 to March 2015 were chosen as study objects, out of them 105 patients were in aute stage and 95 patients were in remisson stage. At same time 100 people from healthy medical examination in our hospital were chosen as control group. The mumber change and subgroups of their peripheral blood NK cells were analyzed and compared.</p><p><b>RESULTS</b>In control group the absolute number of NK cells was (412.91 ± 167.35)/µl, the relative number of NK cells was (13.31 ± 2.56) %; in group at acute stage of leukemia the absolute number of NK cells was (97.84 ± 23.18)/µl, the relative number of NK cells was (6.79 ± 0.78) %; in group at acute stage of lymphoma, the absolute number of NK cells was (101.79 ± 25.63)/µl, and the relative number of NK cells was (7.12 ± 1.03) %; in group at remission stage of leukemia, the absolute number was (297.17 ± 87.56)/µl, and the relative number was (10.15 ± 1.64) %; In group at remission of lymphoma, the absolute number of NK cells was (288.52 ± 118.52)/µl, and the relative number of NK cells was (10.82 ± 1.97) %. The number of NK cells between different groups showed statistical difference (P < 0.05). In remission group, the number of NK cells before and after treatment had statistical difference (P < 0.05). In control group, the number of CD56(bright) subgroup was (25.28 ± 4.72) %, the number of CD56(bright) subgroup at the acute stage of leukemia was (65.46 ± 11.21) %, and the number of CD56(bright) subgroup at the acute stage of lymphoma was (70.71 ± 12.14) %, the number of CD56(bright) subgroup at remission stage of leukemia was (23.35 ± 4.67) %, the number of CD56(bright) subgroup at remission stage of lymphoma was (24.89 ± 4.58) %. The number of CD56(bright) subgroup between different groups showed statistical significance (P < 0.05).</p><p><b>CONCLUSION</b>The number and function of peripheral blood NK cells in patients with hematologic malignancies have been confirmed to be obvious decrement, but after treatment the number of NK cells in those patients showed increment.</p>


Asunto(s)
Humanos , Antígeno CD56 , Neoplasias Hematológicas , Células Asesinas Naturales
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