Objective： To study the characteristics of immunophenotype in acute lymphoblastic leukemia （ALL） and its clinical significance. Methods： Immunophenotyping was performed on 81 ALL patients by three-color flow cytom...Objective： To study the characteristics of immunophenotype in acute lymphoblastic leukemia （ALL） and its clinical significance. Methods： Immunophenotyping was performed on 81 ALL patients by three-color flow cytometry analysis using CD45/SSC gating, meanwhile the cytogenetic analysis was performed on 45 cases out of 81 ALL patients. Results： （1） CD19 was the most commonly expressed of all B-lineage antigens detected with the positive rate being 100%. In T-ALL, the positive expression rate of CD5 and CD7 was the highest, being 90%. Both B-ALL and T-ALL overlapped in expression of lineage antigens. There was no significant difference in the complete remission rate （CR rate） between T-ALL and B-ALL. （2） The incidence of ALL with rayeloid antigens expression （My＋ALL） was 39.5%. CD13 was most often seen among the myeloid markers. My＋ALL always involved in B-lineage antigens and the CR rate in children and adults was 72.2% and 78.6% respectively. （3） The incidence of HAL was 19.8%. Coexpression of B-lineage and myeloid-assoeiated antigens was the commonest subtype in HAL. The expression of CD34 was commonly seen in HAL patients （81.3%）. The CR rate was low in HAL, 50% for children and 40% for adults. （4） Compared to T-ALL, B-ALL, My＋ALL, and HAL had a higher positive rate of CD34 expression with the difference being significant （P〈0.025）. Conclusion： Immunophenotyping had remarkable predominance in diagnosing special category of ALL （such as HAL and My＋ALL）; CD19 and CD5 were highly sensitive in diagnosing B-ALL and T-ALL, but less special, and overlapping was found in expression. No significant association was found between the expression of CD34 or myeloid antigens and CR rate, while low CR rate was found in HAL patients, especially for those coexpressing CD34 antigen.展开更多
The immunophenotyping expression levels of lymphocyte in the peripheral blood from 21 patients with active systemic lupus erythematosus were analyzed by using the immunofluorescence labeling flow cytometry technique...The immunophenotyping expression levels of lymphocyte in the peripheral blood from 21 patients with active systemic lupus erythematosus were analyzed by using the immunofluorescence labeling flow cytometry technique to investigate the immunophenotyping expression of lymphocytes T and B in the peripheral blood of active SLE patients and its clinical value. It was showed that, compared with normal controls, the expression of CD + 3, CD + 4 and the ratio of CD + 4/CD + 8 in the peripheral blood of these patients were decreased , while the expression of CD + 8, CD + 20 was significantly increased . It was suggested that both T and B cells in patients with active SLE involved in immunoregulation, were activated. The abnormal expression of lymphocyte immunophenotyping could influence the immune reaction in SLE patients, which might be one of the important pathogenesis factors in SLE.展开更多
AIM:To assess the capacity to isolate and expand mesenchymal stem cells(MSC)from bone marrow of CBA/Ca,ICR and Balb/c mice. METHODS:Bone marrow of tibia and femur were flushed,cultured and maintained in supplemented D...AIM:To assess the capacity to isolate and expand mesenchymal stem cells(MSC)from bone marrow of CBA/Ca,ICR and Balb/c mice. METHODS:Bone marrow of tibia and femur were flushed,cultured and maintained in supplemented Dulbecco’s modified Eagle’s medium.MSC immunophenotype of cultures were tracked along increasing passages for positivity to CD106,Sca-1 and CD44 and negativity to CD45,CD11b and MHC classⅡ.Differentiation capacity of MSC towards osteogenic and adipo-genic lineages were also assessed. RESULTS:MSC were successfully cultured from bone marrow of all 3 strains,albeit differences in the temporal expression of certain surface antigens.Their differentiation into osteocytes and adipocytes were also observed. MSC from all 3 mouse strains demonstrated a shift from a haematopoietic phenotype(CD106-CD45+CD11b+Sca-1low)to typical MSC phenotype(CD106+CD45-CD11b-Sca-1high)with increasing passages. CONCLUSION:Information garnered assists us in the decision of selecting a mouse strain to generate MSC from for downstream experimentation.展开更多
Glioblastoma-multiforme(GBM), the most aggressive glial tumor, has a worldwide age-adjusted incidence ranging from 0.59-3.69/100000 persons. Despite current multimodal-treatment approach, median-survival time and prog...Glioblastoma-multiforme(GBM), the most aggressive glial tumor, has a worldwide age-adjusted incidence ranging from 0.59-3.69/100000 persons. Despite current multimodal-treatment approach, median-survival time and progression-free survival(PFS) remains short. Glioblastomas display a variety of molecular alterations, which necessitates determining which of these have a prognostic significance. This is a case of a 45-yearold patient who presented with progressive slurring of speech and features of raised intracranial pressure. Computed tomography(CT) scan revealed a large heterogeneously enhancing lesion in the left front-temporalperisylvian region with solid, cystic areas, suggestive of malignant glioma. Partial tumor-excision was followed by concurrent chemo-radiotherapy. Histopathologically, the tumor was astrocytoma grade-IV. Patient had an extended PFS of 12 mo, with an overall survival of 26 mo. Primary-GBM was confirmed using molecular markers and the immunophenotypic signature was defined by evaluating systemic expression of human telomerase reverse transcriptase, interleukin-6, neutrophil-lymphocyte ratio, tissue inhibitor of metalloproteinases-1, human chitinase-3-like-protein-1(YKL-40) and high mobility group-A1. Current findings suggest that this signature can identify worst outcomes, independent of clinical criteria.展开更多
AIM:To study the immunophenotype of hematopoietic progenitor cells from cord blood (CB) grafts (n = 39) in comparison with adult apheresis grafts (AG, n = 229) and pre-apheresis peripheral blood (PAPB) samples (n = 90...AIM:To study the immunophenotype of hematopoietic progenitor cells from cord blood (CB) grafts (n = 39) in comparison with adult apheresis grafts (AG, n = 229) and pre-apheresis peripheral blood (PAPB) samples (n = 908) using flow cytometry analysis.METHODS: First, we performed a qualitative analysis of CD34+ cell sub-populations in both CB and PAPB grafts using the standardized ISHAGE protocol and a wide panel of 20 monoclonal antibodies. Next, we stud-ied some parameters, such as the age of mothers and the weight of newborns, which can influence the qual-ity and the quantity of CD34+ cells from CB. RESULTS: We found that the percentage of apoptotic cells was high in CB in comparison to PAPB (PAPB: 4.6% ± 2.6% vs CB: 53.4% ± 5.2%, P < 0.001). In CB, the weight of newborn and the age of the mother have the influence on CD34+ cells. The follow-up of Ag CD133in the ISHAGE double platform protocol in association with CD45, CD34 and the 7’AAD shows an equal rate between the two cell populations CD133+CD45+CD34+ high and CD34+CD45+ high with a higher percentage. So, is the inclusion of Ac CD133 necessary in the pres-ent panel included in the ISHAGE methodflLast part, we showed a signif icant presence of interferon γ in CB in comparison to PAPB, the annexin showing the high number of apoptotic cells in CB. CONCLUSION: This study demonstrates that many different obstetric factors must be taken into account when processing and cryo-banking umbilical CB units for transplantation.展开更多
In order to study the effects of 1-(2,6-dimethylphenoxy)-2-(3,4-dimethoxyphenylethylamino) propane hydrochloride (DDPH) on proliferation and immunophenotypes of newborn rat pulmonary vascular pericytes induced by hypo...In order to study the effects of 1-(2,6-dimethylphenoxy)-2-(3,4-dimethoxyphenylethylamino) propane hydrochloride (DDPH) on proliferation and immunophenotypes of newborn rat pulmonary vascular pericytes induced by hypoxic endothelial cell conditioned medium (HECCM) from porcine pulmonary arteries, the cultured pericytes were divided into 4 groups according to the endothelial cell conditioned medium (ECCM) used: normoxic ECCM (NECCM) group, NECCM+DDPH group, HECCM group and HECCM+DDPH group. Cell culture, immunocytochemical staining, image analysis and flow cytometric method were used to investigate the effects of HECCM and DDPH on the expression of α-smooth muscle actin (α-SM-Actin) antigen, CD34 antigen, S-100 antigen and proliferating cell nuclear antigen (PCNA) and cell cycle in pericytes. The results showed that the α-SM-Actin antigen in the pericytes in HECCM group was stronger positively expressed than in the other three groups, but CD34 antigen and S-100 antigen were negatively expressed. The expression of α-SM-Actin antigen, CD34 antigen and S-100 antigen was positive in the groups of NECCM, NECCM+DDPH and HECCM+DDPH; The expression of α-SM-Actin and PCNA in HECCM group was 1.32 times (P<0.01) and 1.24 times (P<0.05) that in NECCM group, 1.30 times (P<0.01) and 1.21 times (P<0.05) that in HECCM+DDPH group, respectively. The percentage of the cells in the GO-G1 phase in the HECCM group was lower by 11.7 % and 9.1 %, in S phase higher by 5.6 % and 4.2 %, in G2-M phase higher by 6.1 % and 4.9 % than in the groups of NECCM,HECCM+DDPH, respectively. The inhibitory rate of DDPH on the increased α-SM-Actin and PCNA syntheses in pericytes induced by HECCM were 23.4 % and 17.1 % respectively. The inhibitory rate on the increased pericytes from GO-G1 phase to S phase was 8.3 %. These results suggest that DDPH can directly inhibit pericytes from proliferation and immunophenotypical transformation of smooth muscle-like cells induced by HECCM.展开更多
To investigate the immunophenotypings of malignant epithelial mesothelioma (MEM), and to seek the valuable markers in distinguishing peritoneal MEM from peritoneal metastatic ovarian adenocarcinoma (OA) and colorectal...To investigate the immunophenotypings of malignant epithelial mesothelioma (MEM), and to seek the valuable markers in distinguishing peritoneal MEM from peritoneal metastatic ovarian adenocarcinoma (OA) and colorectal adenocarcinoma (CA), immunohistochemical SP method was used to detect expressions of HBME-1, E-cadherin, CA19-9, MOC-31 and CK7 in paraffin-embedded tissues of 18 cases of MEM, 20 OA and 20 CA. The results showed that there was a significant difference in the expressions of E-cadherin, CA19-9 and MOC-31 between MEM and OA group (P<0.05). Similarly, the difference in the expression of HBME-1, E-cadherin, CA19-9, MOC-31 and CK7 between MEM and CA groups is significant (P<0.05). These results indicate that HBME-1 could be used as a positive marker in distinguishing MEM from CA. E-cadherin, CA19-9 and MOC-31 are considered to be useful negative markers in diagnostic distinction between MEM and metastatic adenocarcinomas, including OA and CA. CK7 is the best positive marker in distinguishing MEM from CA, but this marker appears to be valueless in discriminating MEM from OA.展开更多
To identify the knowledge of rare lymphoproliferative disorder, the clinical and biological features of three kinds of lymphoproliferative disorders with cytoplasmic projections were compared The clinical manifestat...To identify the knowledge of rare lymphoproliferative disorder, the clinical and biological features of three kinds of lymphoproliferative disorders with cytoplasmic projections were compared The clinical manifestations, ultrastructure and immunophenotype were analyzed The results showed that hairy cell leukemia (HCL), splenic lymphoma with villous lymphocyte (SLVL) and hairy cell leukemia-variant (HCL-V) had some common characters including splenomegaly, peripheral blood and bone marrow infiltration by villous lymphocyte and B lymphocyte immunophenotype; but these three disorders had specific features respectively It was concluded that overall analysis of clinical and laboratory features might be contributive to the differential diagnosis of these three disorders展开更多
BACKGROUND Multipotent mesenchymal stromal cells(MSCs)are widely used in the clinic due to their unique properties,namely,their ability to differentiate in all mesenchymal directions and their immunomodulatory activit...BACKGROUND Multipotent mesenchymal stromal cells(MSCs)are widely used in the clinic due to their unique properties,namely,their ability to differentiate in all mesenchymal directions and their immunomodulatory activity.Healthy donor MSCs were used to prevent the development of acute graft vs host disease(GVHD)after allogeneic bone marrow transplantation(allo-BMT).The administration of MSCs to patients was not always effective.The MSCs obtained from different donors have individual characteristics.The differences between MSC samples may affect their clinical efficacy.AIM To study the differences between effective and ineffective MSCs.METHODS MSCs derived from the bone marrow of a hematopoietic stem cells donor were injected intravenously into allo-BMT recipients for GVHD prophylaxis at the moment of blood cell reconstitution.Aliquots of 52 MSC samples that were administered to patients were examined,and the same cells were cultured in the presence of peripheral blood mononuclear cells(PBMCs)from a third-party donor or treated with the pro-inflammatory cytokines IL-1β,IFN and TNF.Flow cytometry revealed the immunophenotype of the nontreated MSCs,the MSCs cocultured with PBMCs for 4 d and the MSCs exposed to cytokines.The proportions of CD25-,CD146-,CD69-,HLA-DR-and PD-1-positive CD4+and CD8+cells and the distribution of various effector and memory cell subpopulations in the PBMCs cocultured with the MSCs were also determined.RESULTS Differences in the immunophenotypes of effective and ineffective MSCs were observed.In the effective samples,the mean fluorescence intensity(MFI)of HLAABC,HLA-DR,CD105,and CD146 was significantly higher.After MSCs were treated with IFN or cocultured with PBMCs,the HLA-ABC,HLA-DR,CD90 and CD54 MFI showed a stronger increase in the effective MSCs,which indicated an increase in the immunomodulatory activity of these cells.When PBMCs were cocultured with effective MSCs,the proportions of CD4+and CD8+central memory cells significantly decreased,and the proportion of CD8+CD146+lymphocytes increased more than in the subpopulations of lymphocytes cocultured with MSC samples that were ineffective in the prevention of GVHD;in addition,the proportion of CD8+effector memory lymphocytes decreased in the PBMCs cocultured with the effective MSC samples but increased in the PBMCs cocultured with the ineffective MSC samples.The proportion of CD4+CD146+lymphocytes increased only when cocultured with the inefficient samples.CONCLUSION For the first time,differences were observed between MSC samples that were effective for GVHD prophylaxis and those that were ineffective.Thus,it was shown that the immunomodulatory activity of MSCs depends on the individual characteristics of the MSC population.展开更多
Monoclonal antibodies were used to label cutaneous germinal center cell-derived lymphomas <CGCCL) obtained from 10 patients. According to the Kiel classification, they were classified into 2 types. Eight patients h...Monoclonal antibodies were used to label cutaneous germinal center cell-derived lymphomas <CGCCL) obtained from 10 patients. According to the Kiel classification, they were classified into 2 types. Eight patients had centroblastic/centrocytic <CB/CC) lymphomas while 2 patients and centrocytic (CC) lymphomas. After monoclonal antibody labelling, the results were consistent with those of the clinical and morphologic analyses. Of the 10 cases, 9 were B1 positive, 6 were K positive, and 4 were λ positive. In 8 cases labeled with immunoglobulin, 6 were IgGFab positive, 2 were IgM positive and 8 were IgA negative. Five cases (CB/CC 3, CC 2) were both Bl, K and IgG positive (γ K). Four cases CB/CC were both Bl and A positive. Only one case (CB/CC) was both K and IgM positive (μ K). Two cases (CB CC) were both A and IgG positive (γ λ). The results indicate that Bl, K and A are the most important markers to phenotype cutaneous B-cell lymphomas. Our findings also show a higher percentage of y K types in CGCCL as compared with Western countries.展开更多
[Objectives]This study was conducted to establish a method for detecting the immunophenotypes of venous blood CIK cells in healthy donors and patients with triple-negative breast cancer or lung cancer by flow cytometr...[Objectives]This study was conducted to establish a method for detecting the immunophenotypes of venous blood CIK cells in healthy donors and patients with triple-negative breast cancer or lung cancer by flow cytometry,and to further analyze and discuss the proportion of cellular immunophenotypes such as CD3^(+),CD4^(+),CD8^(+),CD3^(+)CD8^(+),CD3^(+)CD56^(+)in CIK cells.[Methods]Human venous blood was drawn,then anticoagulated with heparin and isolated with lymphocyte isolation solution,and the relevant immunophenotypes were detected by flow cytometry after 21 d of culture.[Results]The expression of CD3^(+)CD8^(+)and CD3^(+)CD56^(+)in the venous blood CIK cells was significantly higher in healthy donors than that in triple-negative breast and lung cancer patients.[Conclusions]CD3^(+)CD8^(+)and CD3^(+)CD56^(+)CIK cells have high anti-tumor activity.展开更多
Introduction: Diffuse large B-cell lymphoma (DLBCL), not otherwise specified, is a large B-cell lymphoma with a diffuse growth pattern and aggressive clinical course. It is divided in subgroups according to its morpho...Introduction: Diffuse large B-cell lymphoma (DLBCL), not otherwise specified, is a large B-cell lymphoma with a diffuse growth pattern and aggressive clinical course. It is divided in subgroups according to its morphology, immunophenotype, and primary site. Dissemination to bone marrow occurs in 11% to 35% of cases and can be of concordant or discordant morphology. Objective: To examine the association, the type of bone marrow involvement in relation to the primary site, morphology, immunohistochemistry of DLBCLs and to determine the cases of Epstein-Barr virus positive DLBCLs. Materials and Methods: We reviewed lymph node and extranodal biopsies as well as the respective bone marrow biopsies in all cases of DLBCL diagnosed in the Hospital General de México during the period from 2002 to 2010. We used immunohystochemistry for immunophenotype identification (Hans’s algorithm) and an in-situ hybridization technique to detect presence of Epstein Barr encoded RNA (EBER). Results: We included 108 patients with a mean age of 51.9 years, 59 (55%) were men. DLBCL involved lymph nodes in 60% of cases and palatine tonsils in 13%. The centroblastic variant predominated (80%) and 58% originated from activated B-cells. Infiltration of bone marrow was present in 30% of cases and was discordant in 55% of these cases. Correlation between morphology and bone marrow infiltration was statistically significant (P = 0.0003). Presence of Epstein-Barr virus was demonstrated in 15% of patients older than 50 years. Conclusions: Dissemination to bone marrow occurred in 30% of cases and discordant involvement was most common. DLBCL originating from activated B-lymphocytes predominated and the most common extranodal sites were palatine tonsils, suggesting that our population has a clinical behavior similar to Asiatic populations.展开更多
cytometric immunophenotyping has evolved from two-parameter quantitative measurement of peripheral blood lymphocytes to five-or more parameter qualitative evaluation of bone marrow for hematopathology.Leukemia/lymphom...cytometric immunophenotyping has evolved from two-parameter quantitative measurement of peripheral blood lymphocytes to five-or more parameter qualitative evaluation of bone marrow for hematopathology.Leukemia/lymphoma immunophenotyping represent an important addition to histomorphology in the diagnosis,classification and monitoring of hematolymphoid neoplasms.The complexity of five-or more parameter analyses and the interpretation of the data rely on standardization and validation of the instrument,the reagent and the procedure.In addition,clinical flow cytometry laboratories in U.S.are required to document proficiency testing,sample preparation,method accuracy,specificity,sensitivity and precision.CLSI and the U.S.-Canadian Consensus Conference have provided recommendations,but each laboratory is responsible for validating its own qualitative and quantitative procedures.This paper introduces the procedures for quality control of all levels of the operation in a clinical flow cytometry laboratory in USA.展开更多
During B-cell development and maturation,the antigen receptor,which is encoded by the immunoglobulin heavy-(IgH) and light-chain genes,rearrange to associate one of a number of variable,diverse,and joining gene segmen...During B-cell development and maturation,the antigen receptor,which is encoded by the immunoglobulin heavy-(IgH) and light-chain genes,rearrange to associate one of a number of variable,diverse,and joining gene segments.A single mature B-cell expresses an IgH chain and either a kappa or lambda light chain,which is known as allelic or isotypic exclusion.In normal or reactive conditions,lymphoid cells comprise the mixtures of lymphocytes with either kappa or lambda expression.展开更多
To investigate the changes on the immunopbenotypes and the clinical effects of treatment of the late cancer patients with infusion of human peripheral blood lymphocytes stimulated by anti-CD28 and anti-CD80 monoclonal...To investigate the changes on the immunopbenotypes and the clinical effects of treatment of the late cancer patients with infusion of human peripheral blood lymphocytes stimulated by anti-CD28 and anti-CD80 monoclonal antibodies in combination with radiotherapy and chemotherapy, 42 patients with late cancers were collected for study, among which 22 patients were treated with infusion of stimulated lymphocytes in combination with radiotherapy and chemotherapy. The immunological treatment procedure was given twice per week, and one course of treatment consisted of 8 times of giving infusion of lymphocytes. Another 20 patients were selected for control group, in which only radiotherapy and chemotherapy were given without lymphocyte infusions. Flow cytometry was used to examine the immunophenotypes and the clinical symptoms were observed before and after treatments. It was found that the numbers of the CD3^ ＋ , CD4^＋ cells increased, while those of the CD8 ^＋ cells decreased, with an increase of CD4/CD8 radios, but no significant difference existed in case of 22 patients treated with lymphocyte infusion as well as with radiotherapy and chemotherapy. Fifteen patients out of these 22 cases （68.18%）, the immunophenotypes changed obviously with increased numbers of CD3^ ＋ , CD4^ ＋ cells in comparison with those before treatment, and the number of CD95^ ＋ cells was increased after treatment. The PS value in this group of patients decreased after treatment. In comparison with 20 cases in the control group, the immunophenotypes showed no differences before and after treatment. While the PS value decreased obviously. Seven out of the 22 cases （31.83 % ） treated with lymphocyte infusions as well as with radiotherapy and chemotherapy illustrated no major changes in their i mmunophenotypes, compared with the situation before treatment, but the PS value also decreased. In case of treatment with lymphocyte infusions in combination with radiotherapy and chemotherapy, the alteration of phenotypes was reversely correlated with the changes of clinical grades. Although there were 7 cases showing no major alterations of the immunological phenotypes, but their correlation was still evident. In the control group, neither alteration of immunophenotypes nor changes in clinical grades was found. It is concluded that immunotherapy in combination with radiotherapy and chemotherapy can relieve the side effects induced by radiotherapy and chemotherapy and also enhance the therapeutic efforts.展开更多
文摘Objective： To study the characteristics of immunophenotype in acute lymphoblastic leukemia （ALL） and its clinical significance. Methods： Immunophenotyping was performed on 81 ALL patients by three-color flow cytometry analysis using CD45/SSC gating, meanwhile the cytogenetic analysis was performed on 45 cases out of 81 ALL patients. Results： （1） CD19 was the most commonly expressed of all B-lineage antigens detected with the positive rate being 100%. In T-ALL, the positive expression rate of CD5 and CD7 was the highest, being 90%. Both B-ALL and T-ALL overlapped in expression of lineage antigens. There was no significant difference in the complete remission rate （CR rate） between T-ALL and B-ALL. （2） The incidence of ALL with rayeloid antigens expression （My＋ALL） was 39.5%. CD13 was most often seen among the myeloid markers. My＋ALL always involved in B-lineage antigens and the CR rate in children and adults was 72.2% and 78.6% respectively. （3） The incidence of HAL was 19.8%. Coexpression of B-lineage and myeloid-assoeiated antigens was the commonest subtype in HAL. The expression of CD34 was commonly seen in HAL patients （81.3%）. The CR rate was low in HAL, 50% for children and 40% for adults. （4） Compared to T-ALL, B-ALL, My＋ALL, and HAL had a higher positive rate of CD34 expression with the difference being significant （P〈0.025）. Conclusion： Immunophenotyping had remarkable predominance in diagnosing special category of ALL （such as HAL and My＋ALL）; CD19 and CD5 were highly sensitive in diagnosing B-ALL and T-ALL, but less special, and overlapping was found in expression. No significant association was found between the expression of CD34 or myeloid antigens and CR rate, while low CR rate was found in HAL patients, especially for those coexpressing CD34 antigen.
文摘The immunophenotyping expression levels of lymphocyte in the peripheral blood from 21 patients with active systemic lupus erythematosus were analyzed by using the immunofluorescence labeling flow cytometry technique to investigate the immunophenotyping expression of lymphocytes T and B in the peripheral blood of active SLE patients and its clinical value. It was showed that, compared with normal controls, the expression of CD + 3, CD + 4 and the ratio of CD + 4/CD + 8 in the peripheral blood of these patients were decreased , while the expression of CD + 8, CD + 20 was significantly increased . It was suggested that both T and B cells in patients with active SLE involved in immunoregulation, were activated. The abnormal expression of lymphocyte immunophenotyping could influence the immune reaction in SLE patients, which might be one of the important pathogenesis factors in SLE.
基金Supported by The Research University Grant Scheme UPM,04-02-10-0924RUExploratory Research Grant Scheme,Ministry of Higher Education,ERGS/1/2012/5527106
文摘AIM:To assess the capacity to isolate and expand mesenchymal stem cells(MSC)from bone marrow of CBA/Ca,ICR and Balb/c mice. METHODS:Bone marrow of tibia and femur were flushed,cultured and maintained in supplemented Dulbecco’s modified Eagle’s medium.MSC immunophenotype of cultures were tracked along increasing passages for positivity to CD106,Sca-1 and CD44 and negativity to CD45,CD11b and MHC classⅡ.Differentiation capacity of MSC towards osteogenic and adipo-genic lineages were also assessed. RESULTS:MSC were successfully cultured from bone marrow of all 3 strains,albeit differences in the temporal expression of certain surface antigens.Their differentiation into osteocytes and adipocytes were also observed. MSC from all 3 mouse strains demonstrated a shift from a haematopoietic phenotype(CD106-CD45+CD11b+Sca-1low)to typical MSC phenotype(CD106+CD45-CD11b-Sca-1high)with increasing passages. CONCLUSION:Information garnered assists us in the decision of selecting a mouse strain to generate MSC from for downstream experimentation.
基金Supported by M.P.Biotech Council,M.P.for financial assistance and BMHRC for infrastructural facilities,No.249
文摘Glioblastoma-multiforme(GBM), the most aggressive glial tumor, has a worldwide age-adjusted incidence ranging from 0.59-3.69/100000 persons. Despite current multimodal-treatment approach, median-survival time and progression-free survival(PFS) remains short. Glioblastomas display a variety of molecular alterations, which necessitates determining which of these have a prognostic significance. This is a case of a 45-yearold patient who presented with progressive slurring of speech and features of raised intracranial pressure. Computed tomography(CT) scan revealed a large heterogeneously enhancing lesion in the left front-temporalperisylvian region with solid, cystic areas, suggestive of malignant glioma. Partial tumor-excision was followed by concurrent chemo-radiotherapy. Histopathologically, the tumor was astrocytoma grade-IV. Patient had an extended PFS of 12 mo, with an overall survival of 26 mo. Primary-GBM was confirmed using molecular markers and the immunophenotypic signature was defined by evaluating systemic expression of human telomerase reverse transcriptase, interleukin-6, neutrophil-lymphocyte ratio, tissue inhibitor of metalloproteinases-1, human chitinase-3-like-protein-1(YKL-40) and high mobility group-A1. Current findings suggest that this signature can identify worst outcomes, independent of clinical criteria.
文摘AIM:To study the immunophenotype of hematopoietic progenitor cells from cord blood (CB) grafts (n = 39) in comparison with adult apheresis grafts (AG, n = 229) and pre-apheresis peripheral blood (PAPB) samples (n = 908) using flow cytometry analysis.METHODS: First, we performed a qualitative analysis of CD34+ cell sub-populations in both CB and PAPB grafts using the standardized ISHAGE protocol and a wide panel of 20 monoclonal antibodies. Next, we stud-ied some parameters, such as the age of mothers and the weight of newborns, which can influence the qual-ity and the quantity of CD34+ cells from CB. RESULTS: We found that the percentage of apoptotic cells was high in CB in comparison to PAPB (PAPB: 4.6% ± 2.6% vs CB: 53.4% ± 5.2%, P < 0.001). In CB, the weight of newborn and the age of the mother have the influence on CD34+ cells. The follow-up of Ag CD133in the ISHAGE double platform protocol in association with CD45, CD34 and the 7’AAD shows an equal rate between the two cell populations CD133+CD45+CD34+ high and CD34+CD45+ high with a higher percentage. So, is the inclusion of Ac CD133 necessary in the pres-ent panel included in the ISHAGE methodflLast part, we showed a signif icant presence of interferon γ in CB in comparison to PAPB, the annexin showing the high number of apoptotic cells in CB. CONCLUSION: This study demonstrates that many different obstetric factors must be taken into account when processing and cryo-banking umbilical CB units for transplantation.
文摘In order to study the effects of 1-(2,6-dimethylphenoxy)-2-(3,4-dimethoxyphenylethylamino) propane hydrochloride (DDPH) on proliferation and immunophenotypes of newborn rat pulmonary vascular pericytes induced by hypoxic endothelial cell conditioned medium (HECCM) from porcine pulmonary arteries, the cultured pericytes were divided into 4 groups according to the endothelial cell conditioned medium (ECCM) used: normoxic ECCM (NECCM) group, NECCM+DDPH group, HECCM group and HECCM+DDPH group. Cell culture, immunocytochemical staining, image analysis and flow cytometric method were used to investigate the effects of HECCM and DDPH on the expression of α-smooth muscle actin (α-SM-Actin) antigen, CD34 antigen, S-100 antigen and proliferating cell nuclear antigen (PCNA) and cell cycle in pericytes. The results showed that the α-SM-Actin antigen in the pericytes in HECCM group was stronger positively expressed than in the other three groups, but CD34 antigen and S-100 antigen were negatively expressed. The expression of α-SM-Actin antigen, CD34 antigen and S-100 antigen was positive in the groups of NECCM, NECCM+DDPH and HECCM+DDPH; The expression of α-SM-Actin and PCNA in HECCM group was 1.32 times (P<0.01) and 1.24 times (P<0.05) that in NECCM group, 1.30 times (P<0.01) and 1.21 times (P<0.05) that in HECCM+DDPH group, respectively. The percentage of the cells in the GO-G1 phase in the HECCM group was lower by 11.7 % and 9.1 %, in S phase higher by 5.6 % and 4.2 %, in G2-M phase higher by 6.1 % and 4.9 % than in the groups of NECCM,HECCM+DDPH, respectively. The inhibitory rate of DDPH on the increased α-SM-Actin and PCNA syntheses in pericytes induced by HECCM were 23.4 % and 17.1 % respectively. The inhibitory rate on the increased pericytes from GO-G1 phase to S phase was 8.3 %. These results suggest that DDPH can directly inhibit pericytes from proliferation and immunophenotypical transformation of smooth muscle-like cells induced by HECCM.
文摘To investigate the immunophenotypings of malignant epithelial mesothelioma (MEM), and to seek the valuable markers in distinguishing peritoneal MEM from peritoneal metastatic ovarian adenocarcinoma (OA) and colorectal adenocarcinoma (CA), immunohistochemical SP method was used to detect expressions of HBME-1, E-cadherin, CA19-9, MOC-31 and CK7 in paraffin-embedded tissues of 18 cases of MEM, 20 OA and 20 CA. The results showed that there was a significant difference in the expressions of E-cadherin, CA19-9 and MOC-31 between MEM and OA group (P<0.05). Similarly, the difference in the expression of HBME-1, E-cadherin, CA19-9, MOC-31 and CK7 between MEM and CA groups is significant (P<0.05). These results indicate that HBME-1 could be used as a positive marker in distinguishing MEM from CA. E-cadherin, CA19-9 and MOC-31 are considered to be useful negative markers in diagnostic distinction between MEM and metastatic adenocarcinomas, including OA and CA. CK7 is the best positive marker in distinguishing MEM from CA, but this marker appears to be valueless in discriminating MEM from OA.
文摘To identify the knowledge of rare lymphoproliferative disorder, the clinical and biological features of three kinds of lymphoproliferative disorders with cytoplasmic projections were compared The clinical manifestations, ultrastructure and immunophenotype were analyzed The results showed that hairy cell leukemia (HCL), splenic lymphoma with villous lymphocyte (SLVL) and hairy cell leukemia-variant (HCL-V) had some common characters including splenomegaly, peripheral blood and bone marrow infiltration by villous lymphocyte and B lymphocyte immunophenotype; but these three disorders had specific features respectively It was concluded that overall analysis of clinical and laboratory features might be contributive to the differential diagnosis of these three disorders
文摘BACKGROUND Multipotent mesenchymal stromal cells(MSCs)are widely used in the clinic due to their unique properties,namely,their ability to differentiate in all mesenchymal directions and their immunomodulatory activity.Healthy donor MSCs were used to prevent the development of acute graft vs host disease(GVHD)after allogeneic bone marrow transplantation(allo-BMT).The administration of MSCs to patients was not always effective.The MSCs obtained from different donors have individual characteristics.The differences between MSC samples may affect their clinical efficacy.AIM To study the differences between effective and ineffective MSCs.METHODS MSCs derived from the bone marrow of a hematopoietic stem cells donor were injected intravenously into allo-BMT recipients for GVHD prophylaxis at the moment of blood cell reconstitution.Aliquots of 52 MSC samples that were administered to patients were examined,and the same cells were cultured in the presence of peripheral blood mononuclear cells(PBMCs)from a third-party donor or treated with the pro-inflammatory cytokines IL-1β,IFN and TNF.Flow cytometry revealed the immunophenotype of the nontreated MSCs,the MSCs cocultured with PBMCs for 4 d and the MSCs exposed to cytokines.The proportions of CD25-,CD146-,CD69-,HLA-DR-and PD-1-positive CD4+and CD8+cells and the distribution of various effector and memory cell subpopulations in the PBMCs cocultured with the MSCs were also determined.RESULTS Differences in the immunophenotypes of effective and ineffective MSCs were observed.In the effective samples,the mean fluorescence intensity(MFI)of HLAABC,HLA-DR,CD105,and CD146 was significantly higher.After MSCs were treated with IFN or cocultured with PBMCs,the HLA-ABC,HLA-DR,CD90 and CD54 MFI showed a stronger increase in the effective MSCs,which indicated an increase in the immunomodulatory activity of these cells.When PBMCs were cocultured with effective MSCs,the proportions of CD4+and CD8+central memory cells significantly decreased,and the proportion of CD8+CD146+lymphocytes increased more than in the subpopulations of lymphocytes cocultured with MSC samples that were ineffective in the prevention of GVHD;in addition,the proportion of CD8+effector memory lymphocytes decreased in the PBMCs cocultured with the effective MSC samples but increased in the PBMCs cocultured with the ineffective MSC samples.The proportion of CD4+CD146+lymphocytes increased only when cocultured with the inefficient samples.CONCLUSION For the first time,differences were observed between MSC samples that were effective for GVHD prophylaxis and those that were ineffective.Thus,it was shown that the immunomodulatory activity of MSCs depends on the individual characteristics of the MSC population.
文摘Monoclonal antibodies were used to label cutaneous germinal center cell-derived lymphomas <CGCCL) obtained from 10 patients. According to the Kiel classification, they were classified into 2 types. Eight patients had centroblastic/centrocytic <CB/CC) lymphomas while 2 patients and centrocytic (CC) lymphomas. After monoclonal antibody labelling, the results were consistent with those of the clinical and morphologic analyses. Of the 10 cases, 9 were B1 positive, 6 were K positive, and 4 were λ positive. In 8 cases labeled with immunoglobulin, 6 were IgGFab positive, 2 were IgM positive and 8 were IgA negative. Five cases (CB/CC 3, CC 2) were both Bl, K and IgG positive (γ K). Four cases CB/CC were both Bl and A positive. Only one case (CB/CC) was both K and IgM positive (μ K). Two cases (CB CC) were both A and IgG positive (γ λ). The results indicate that Bl, K and A are the most important markers to phenotype cutaneous B-cell lymphomas. Our findings also show a higher percentage of y K types in CGCCL as compared with Western countries.
基金Supported by The 2021 Undergraduate Innovation and Entrepreneurship Training Program of Guangdong Province(S202110564094)Horizontal Project of South China Agricultural University(h20220138).
文摘[Objectives]This study was conducted to establish a method for detecting the immunophenotypes of venous blood CIK cells in healthy donors and patients with triple-negative breast cancer or lung cancer by flow cytometry,and to further analyze and discuss the proportion of cellular immunophenotypes such as CD3^(+),CD4^(+),CD8^(+),CD3^(+)CD8^(+),CD3^(+)CD56^(+)in CIK cells.[Methods]Human venous blood was drawn,then anticoagulated with heparin and isolated with lymphocyte isolation solution,and the relevant immunophenotypes were detected by flow cytometry after 21 d of culture.[Results]The expression of CD3^(+)CD8^(+)and CD3^(+)CD56^(+)in the venous blood CIK cells was significantly higher in healthy donors than that in triple-negative breast and lung cancer patients.[Conclusions]CD3^(+)CD8^(+)and CD3^(+)CD56^(+)CIK cells have high anti-tumor activity.
文摘Introduction: Diffuse large B-cell lymphoma (DLBCL), not otherwise specified, is a large B-cell lymphoma with a diffuse growth pattern and aggressive clinical course. It is divided in subgroups according to its morphology, immunophenotype, and primary site. Dissemination to bone marrow occurs in 11% to 35% of cases and can be of concordant or discordant morphology. Objective: To examine the association, the type of bone marrow involvement in relation to the primary site, morphology, immunohistochemistry of DLBCLs and to determine the cases of Epstein-Barr virus positive DLBCLs. Materials and Methods: We reviewed lymph node and extranodal biopsies as well as the respective bone marrow biopsies in all cases of DLBCL diagnosed in the Hospital General de México during the period from 2002 to 2010. We used immunohystochemistry for immunophenotype identification (Hans’s algorithm) and an in-situ hybridization technique to detect presence of Epstein Barr encoded RNA (EBER). Results: We included 108 patients with a mean age of 51.9 years, 59 (55%) were men. DLBCL involved lymph nodes in 60% of cases and palatine tonsils in 13%. The centroblastic variant predominated (80%) and 58% originated from activated B-cells. Infiltration of bone marrow was present in 30% of cases and was discordant in 55% of these cases. Correlation between morphology and bone marrow infiltration was statistically significant (P = 0.0003). Presence of Epstein-Barr virus was demonstrated in 15% of patients older than 50 years. Conclusions: Dissemination to bone marrow occurred in 30% of cases and discordant involvement was most common. DLBCL originating from activated B-lymphocytes predominated and the most common extranodal sites were palatine tonsils, suggesting that our population has a clinical behavior similar to Asiatic populations.
文摘cytometric immunophenotyping has evolved from two-parameter quantitative measurement of peripheral blood lymphocytes to five-or more parameter qualitative evaluation of bone marrow for hematopathology.Leukemia/lymphoma immunophenotyping represent an important addition to histomorphology in the diagnosis,classification and monitoring of hematolymphoid neoplasms.The complexity of five-or more parameter analyses and the interpretation of the data rely on standardization and validation of the instrument,the reagent and the procedure.In addition,clinical flow cytometry laboratories in U.S.are required to document proficiency testing,sample preparation,method accuracy,specificity,sensitivity and precision.CLSI and the U.S.-Canadian Consensus Conference have provided recommendations,but each laboratory is responsible for validating its own qualitative and quantitative procedures.This paper introduces the procedures for quality control of all levels of the operation in a clinical flow cytometry laboratory in USA.
文摘During B-cell development and maturation,the antigen receptor,which is encoded by the immunoglobulin heavy-(IgH) and light-chain genes,rearrange to associate one of a number of variable,diverse,and joining gene segments.A single mature B-cell expresses an IgH chain and either a kappa or lambda light chain,which is known as allelic or isotypic exclusion.In normal or reactive conditions,lymphoid cells comprise the mixtures of lymphocytes with either kappa or lambda expression.
文摘To investigate the changes on the immunopbenotypes and the clinical effects of treatment of the late cancer patients with infusion of human peripheral blood lymphocytes stimulated by anti-CD28 and anti-CD80 monoclonal antibodies in combination with radiotherapy and chemotherapy, 42 patients with late cancers were collected for study, among which 22 patients were treated with infusion of stimulated lymphocytes in combination with radiotherapy and chemotherapy. The immunological treatment procedure was given twice per week, and one course of treatment consisted of 8 times of giving infusion of lymphocytes. Another 20 patients were selected for control group, in which only radiotherapy and chemotherapy were given without lymphocyte infusions. Flow cytometry was used to examine the immunophenotypes and the clinical symptoms were observed before and after treatments. It was found that the numbers of the CD3^ ＋ , CD4^＋ cells increased, while those of the CD8 ^＋ cells decreased, with an increase of CD4/CD8 radios, but no significant difference existed in case of 22 patients treated with lymphocyte infusion as well as with radiotherapy and chemotherapy. Fifteen patients out of these 22 cases （68.18%）, the immunophenotypes changed obviously with increased numbers of CD3^ ＋ , CD4^ ＋ cells in comparison with those before treatment, and the number of CD95^ ＋ cells was increased after treatment. The PS value in this group of patients decreased after treatment. In comparison with 20 cases in the control group, the immunophenotypes showed no differences before and after treatment. While the PS value decreased obviously. Seven out of the 22 cases （31.83 % ） treated with lymphocyte infusions as well as with radiotherapy and chemotherapy illustrated no major changes in their i mmunophenotypes, compared with the situation before treatment, but the PS value also decreased. In case of treatment with lymphocyte infusions in combination with radiotherapy and chemotherapy, the alteration of phenotypes was reversely correlated with the changes of clinical grades. Although there were 7 cases showing no major alterations of the immunological phenotypes, but their correlation was still evident. In the control group, neither alteration of immunophenotypes nor changes in clinical grades was found. It is concluded that immunotherapy in combination with radiotherapy and chemotherapy can relieve the side effects induced by radiotherapy and chemotherapy and also enhance the therapeutic efforts.
