RESUMEN
OBJECTIVE: To study the correlation between expression of CD200 and regulatory T cells (Tregs) in multiple myeloma(MM) patients and to explore its significance in prognostic stratification. METHODS: CD200 and other immunophenotypes, including CD38, CD138, CD56, CD19, CD20, CD117, cytoplasm light chain Kappa and Lambda in bone marrow samples, and Tregs in peripheral blood were detected by flow cytometry from 78 newly diagnosed MM patients. Serum concentrations of hemoglobin(Hb), ß2 microglobulin (ß2-MG) and lactate dehydrogenase (LDH) were detected, respectively. The new risk stratification of patients was carried out according to international stage system (ISS) and cytogenetic characteristics. The correlation between expression of CD200 and Tregs in MM patients was analyzed and their differences in prognosis were compared. RESULTS: The positive rate of CD200 expression was 71.79% in 78 patients (56/78). The expression of CD200 in sex and age of patients was no significant different. The expression of CD117 in CD200+ group was significantly higher than that of in CD200- group (P=0.032). There was no significant difference in the expression of CD20, CD56 and CD19 between 2 groups. The level of Hb in CD200+ group was significantly lower than that in CD200- group (P=0.035). The level of ß2-MG in CD200+ group was significantly higher than that in CD200- group (P=0.013). There was no significant difference in the level of LDH between 2 groups. In CD200+ group, 17 patients were in stageâ , accounting for 58.62% (17/29), 30 patients were in stageâ ¡, accounting for 75% (30/40), 9 patients were in stage â ¢, accounting for 100% (9/9). With the increase of CD200 expression intensity, the risk of prognostic stratification went up (P=0). The brighter CD200 expressed, the worse the prognosis was. The percentage of Trges in CD200+ group was significantly higher than that of in CD200- group (P=0.043). The content of Tregs was positively correlated with the expression of CD200 (r=0.743, P=0.044). Overall survival in CD200- group was significantly higher than that in CD200+ group (P=0.036). Progression-free survival in CD200- group was a bit higher than that of in CD200+ group, but it wasn't statistically significant. CONCLUSION: The expression of CD200 positively correlates with the percentage of Tregs in MM patients. It is an important factor of poor prognosis and a reliable indicator for the evaluation by clinical efficacy in MM patients.
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Mieloma Múltiple , Linfocitos T Reguladores , Antígenos CD , Médula Ósea , Supervivencia sin Enfermedad , Citometría de Flujo , Humanos , Cadenas kappa de Inmunoglobulina , Inmunofenotipificación , L-Lactato Deshidrogenasa , Pronóstico , Resultado del TratamientoRESUMEN
OBJECTIVE: To investigate the expression and subcellular distribution of costimulatory molecules B7-H1, B7-H3 and B7-H4 in human hematologic malignancy cell lines. METHODS: The expression and subcellular distribution of B7-H1, B7-H3 and B7-H4 in 13 human hematologic malignancy cell lines were determined by RT-PCR, qPCR, Western blot and flow cytometry, the peripheral blood mononuclear cells (PB MNC) of 12 volunteers were used as control. RESULTS: The mRNA of B7-H1, B7-H3 and B7-H4 was widely expressed in PB MNC and hematologic malignancy cell lines, with a lower level of B7-H4. The mRNA expression of 3 molecules was highest in Maver, Z138, and HL-60, respectively, while among them the B7-H3 and B7-H4 had no expression in CZ1. The nuclear and cytoplasmic protein of 3 costimulatory molecules abnormally overexpressed only in hematologic malignancy cell lines, with the highest level in U937, Z138, and Raji, respectively, while the B7-H3 and B7-H4 had no expression in CZ1. There were differences among mRNA expression, nuclear and cytoplasmic protein expression of 3 molecules in cell lines derived from the same type of tumor, but the differences of expression in mRNA and protein levels were not exactly the same. The B7-H3 expression abundance in membrane localization was higher in U937, Maver and Z138, while the membrane protein of B7-H1 and B7-H4 had no or low expression in 13 cell lines. CONCLUSION: The mRNA expression of costimulatory molecules B7-H1, B7-H3 and B7-H4 can be widely detected. The protein level of 3 costimulatory molecules abnormally overexpressed only in hematologic malignancy cell lines, moreover the subcellular localizations mostly was found in nucleus and cytoplasm, while the membrane protein expresses in low level or had no expression. There are differences among the expression of 3 molecules in cell lines derived from the same type of tumor.
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Neoplasias Hematológicas , Leucocitos Mononucleares , Antígeno B7-H1 , Línea Celular Tumoral , Citoplasma , Citometría de Flujo , HumanosRESUMEN
Thymopentin is a group of biologically active peptide secreted mainly by the epithelial cells of thymic cortex and medulla. Whether it promotes T cells production from human embryonic stem cells(hESCs) in vitro remains an elusive issue. In the present study, we develop a novel strategy that enhances T-cell lineage differentiation of hESCs in collagen matrix culture by sequential cytokine cocktails treatment combined with thymopentin stimulation. We observed that approximately 30.75% cells expressed CD34 on day 14 of the cultures and expressed the surface markers of erythroid, lymphoid and myeloid lineages. The results of colony assays and gene expressions by RT-PCR analysis also demonstrated that hematopoietic progenitor cells (HPCs) derived from hESCs were capable of multi-lineage differentiation. Further study revealed that culturing with thymopentin treatment, the CD34(+)CD45RA(+)CD7(+) cells sorted from HPCs expressed T-cell-related genes, IKAROS, DNTT, TCRγ and TCRß, and T-cell surface markers, CD3, cytoplasmic CD3, CD5, CD27, TCRγδ, CD4 and CD8. The differentiated cells produced the cytokines including IFN-γ, IL-2 and TNF-α in response to stimulation, providing the evidence for T-cell function of these cells. In conclusion, thymopentin enhances T-cell lineage differentiation from hESCs in vitro by mimicking thymus peptide environment in vivo.