Chimeric antigen receptor (CAR) T cell therapy has exhibited dramatic anti-tumor efficacy in clinical trials. In this study, we reported the transcriptome profiles of bone marrow cells in four B cell acute lymphoblastic leukemia (B-ALL) patients before and after CD19-specific CAR-T therapy. CD19-CAR-T therapy remarkably reduced the number of leukemia cells, and three patients achieved bone marrow remission (minimal residual disease negative). The efficacy of CD19-CAR-T therapy on B-ALL was positively correlated with the abundance of CAR and immune cell subpopulations, e.g., CD8 T cells and natural killer (NK) cells, in the bone marrow. Additionally, CD19-CAR-T therapy mainly influenced the expression of genes linked to cell cycle and immune response pathways, including the NK cell mediated cytotoxicity and NOD-like receptor signaling pathways. The regulatory network analyses revealed that microRNAs (e.g., miR-148a-3p and miR-375), acting as oncogenes or tumor suppressors, could regulate the crosstalk between the genes encoding transcription factors (TFs; e.g., JUN and FOS) and histones (e.g., HIST1H4A and HIST2H4A) involved in CD19-CAR-T therapy. Furthermore, many long non-coding RNAs showed a high degree of co-expression with TFs or histones (e.g., FOS and HIST1H4B) and were associated with immune processes. These transcriptome analyses provided important clues for further understanding the gene expression and related mechanisms underlying the efficacy of CAR-T immunotherapy.
Adult ; Antigens, CD19 ; metabolism ; Bone Marrow ; metabolism ; CD8-Positive T-Lymphocytes ; immunology ; Female ; Gene Expression Regulation, Leukemic ; Gene Regulatory Networks ; Humans ; Immunotherapy, Adoptive ; Male ; MicroRNAs ; genetics ; metabolism ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; immunology ; therapy ; RNA, Long Noncoding ; genetics ; metabolism ; Receptors, Antigen, T-Cell ; Transcription Factors ; metabolism ; Transcriptome ; genetics

Acute graft-versus-host disease (aGVHD) is a serious complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT). However, the mechanisms of aGVHD are not well understood. We aim to investigate the roles of the three angiogenic factors: angiopoietin-1 (Ang-1), Ang-2 and vascular endothelial growth factor (VEGF) in the development of aGVHD. Twenty-one patients who underwent allo-HSCT were included in our study. The dynamic changes of Ang-1, Ang-2 and VEGF were monitored in patients before and after allo-HSCT. In vitro, endothelial cells (ECs) were treated with TNF-β in the presence or absence of Ang-1, and then the Ang-2 level in the cell culture medium and the tubule formation by ECs were evaluated. After allo-HSCT, Ang-1, Ang-2 and VEGF all exhibited significant variation, suggesting these factors might be involved in the endothelial damage in transplantation. Patients with aGVHD had lower Ang-1 level at day 7 but higher Ang-2 level at day 21 than those without aGVHD, implying that Ang-1 may play a protective role in early phase yet Ang-2 is a promotion factor to aGVHD. In vitro, TNF-β promoted the release of Ang-2 by ECs and impaired tubule formation of ECs, which were both weakened by Ang-1, suggesting that Ang-1 may play a protective role in aGVHD by influencing the secretion of Ang-2, consistent with our in vivo tests. It is concluded that monitoring changes of these factors following allo-HSCT might help to identify patients at a high risk for aGVHD.
Acute Disease ; Adolescent ; Adult ; Angiogenesis Inducing Agents ; immunology ; metabolism ; pharmacology ; Angiopoietin-1 ; genetics ; immunology ; pharmacology ; Angiopoietin-2 ; genetics ; immunology ; pharmacology ; Antineoplastic Agents ; therapeutic use ; Female ; Gene Expression Regulation, Neoplastic ; Graft vs Host Disease ; genetics ; immunology ; pathology ; Hematopoietic Stem Cell Transplantation ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; immunology ; Humans ; Leukemia, Myeloid ; genetics ; immunology ; pathology ; therapy ; Lymphoma, Non-Hodgkin ; genetics ; immunology ; pathology ; therapy ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; immunology ; pathology ; therapy ; Retrospective Studies ; Signal Transduction ; Transplantation, Homologous ; Tumor Necrosis Factor-alpha ; pharmacology ; Vascular Endothelial Growth Factor A ; genetics ; immunology

No abstract available.
Adolescent ; Antineoplastic Combined Chemotherapy Protocols/therapeutic use ; Burkitt Lymphoma/*pathology ; Child, Preschool ; Cyclophosphamide/therapeutic use ; Doxorubicin/therapeutic use ; Female ; Gene Rearrangement ; Herpesvirus 4, Human/metabolism ; Humans ; Immunohistochemistry ; Lymphoma, B-Cell/*diagnosis/drug therapy ; Lymphoma, Large B-Cell, Diffuse/*pathology ; Male ; Prednisone/therapeutic use ; Proto-Oncogene Proteins c-myc/genetics ; Tomography, X-Ray Computed ; Vincristine/therapeutic use ; Viral Matrix Proteins/immunology/metabolism

No abstract available.
Antibodies, Monoclonal, Murine-Derived/therapeutic use ; Antineoplastic Combined Chemotherapy Protocols/therapeutic use ; B-Cell-Specific Activator Protein/metabolism ; Bone Marrow/metabolism/*pathology ; Cyclophosphamide/therapeutic use ; Doxorubicin/therapeutic use ; Endoscopy, Digestive System ; Female ; Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor ; Genetic Loci ; Humans ; Liver/metabolism/pathology ; Lymphocytes/cytology/immunology ; Lymphoma, Large B-Cell, Diffuse/complications/*diagnosis/drug therapy ; Lymphoma, T-Cell, Peripheral/complications/*diagnosis/drug therapy ; Middle Aged ; Prednisone/therapeutic use ; Receptors, Antigen, T-Cell, gamma-delta/genetics ; Tomography, X-Ray Computed ; Vincristine/therapeutic use

INTRODUCTIONThis study sought to investigate the immunophenotypic subtype profiles of 110 Chinese adult patients with acute lymphoblastic leukaemia (ALL) and its association to cytogenetics and the clinical features.

MATERIALS AND METHODSA total of 110 adult patients with ALL were immunophenotyped by CD45/SSC double parameters and 4 colour flow cytometry. Seventy-three cases were also subjected to karyotype analysis by R-banding technology. The clinical and laboratory data of 110 ALL patients were retrospectively analysed.

RESULTSOf all the patients, 21.8% were identified as T-ALL, 78.2% as B-ALL. Abnormal karyotypes were detected in 37 out of 73 (50.7%) cases and the most common cytogenetic abnormality was the Philadelphia (Ph) chromosome, which was found in 23.3% (17/73) of the cases. Myeloid antigen (MyAg) expression was documented in 47.3% of the 110 adult ALL cases analysed and CD13 was the most commonly expressed MyAg in ALL patients (32.1 %). No difference was observed in the expression of MyAg between the groups of patients with T-ALL (45.8%) and B-ALL (47.7%). Our data showed that older age, higher CD34 positivity and lower proportion of patients with splenomegaly were found to be correlated with MyAg+ ALL, and that patients with Ph+ B-ALL were older, presented with higher haemoglobin level and higher CD34 expression. No statistical difference was noted in complete remission (CR) rate, relapse rate, induction mortality or total death rate among My+ and My-, Ph+ and Ph-, or B-ALL and T-ALL patients.

CONCLUSIONOur results indicate that the distribution of ALL in Chinese adult patients was similar with the general distribution pattern in the other countries, and the expression of MyAg in patients with T-ALL and B-ALL was comparable. Both the expression of MyAg and the presence of Ph chromosome in adult ALL were significantly associated with median age and CD34 expression while not with the response to induction treatment.

Adolescent ; Adult ; Aged ; Asian Continental Ancestry Group ; Cytogenetic Analysis ; Female ; Humans ; Immunophenotyping ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; genetics ; immunology ; Retrospective Studies ; Young Adult

Mantle cell lymphoma (MCL) is a kind of mature B-cell neoplasms with significantly poor prognosis and is usually misdiagnosed. With the development of flow cytometry and cytogenetic technique, most patients were at leukemic phase when diagnosed. This study was purposed to investigate the immunophenotypes of MCL, the immunophenotype information of 22 leukemic MCL patients was analyzed retrospectively. All the patients were conformed t(11;14) translocation by fluorescence in situ hybridization. Immunophenotypes were detected by a four-color flow cytometry including CD3, CD4, CD5, CD8, CD10, CD19, CD20, CD22, CD23, CD25, CD38, CD103, CD148, CD200, FMC7, ZAP-70, κ, λ. The results showed that CD19, CD5, CD20 and monoclonal sIg expressed in all 22 patients with CD20 high expression; CD22 expressed weakly in 17 patients; CD23 expressed in 6 patients including 2 cases highly expressed; FMC7 expressed in 12 patients. 5 patients were 4-point score and 17 patients had a score less than 4 according to CLL scoring system. CD148 and CD200 were detected in 18 patients, in which CD200 expressed negatively in 11 patients, CD200 expressed weakly in 7 patients with median fluorescence intensity (MFI) 25.8 (6.6 - 254.26); CD148 expressed positively in all 18 patients with median MFI: 337 (73.4 - 1341.9). It is concluded that the atypical immunophenotype is common in leukemia MCL, thereby the diagnosis of MCL needs comprehensively analyze with morphocytology, immunophenotype and cytogenetic, CD200 and CD148 as new bio-markers can differentiate MCL from chronic B cell lymphoproliferative disease.
Adult ; Aged ; Aged, 80 and over ; Antigens, CD ; metabolism ; Female ; Humans ; Immunophenotyping ; Karyotyping ; Lymphoma, Mantle-Cell ; genetics ; immunology ; metabolism ; Male ; Middle Aged ; Receptor-Like Protein Tyrosine Phosphatases, Class 3 ; metabolism ; Retrospective Studies

This study was aimed to investigate clinical and prognostic significances of 4 target antigens (CD19, CD20, CD22 and CD33) for antibody-based immunotherapy and to evaluate the applications of these antibody-based target therapy to adult acute lymphoblastic leukemia (ALL). The immunophenotype of 220 adult patients with ALL were analyzed by four-color flow Cytometry, and cytogenetic and molecular parameters were detected by conventional cytogenetics, fluorescence in situ hybridization, real-time quantitative PCR, nested PCR and DNA sequencing. The results showed that CD19 positive (CD19(+)) cases were more in female (46.4% vs. 23.4%, P = 0.006), elderly patients aged > 60 years (14.4% vs. 2.1%, P = 0.022), CD33(+) co-expression cases (47.8% vs. 12.0%, P = 0.001) and genetic high-risk group (55.8% vs. 20.8%, P = 0.002) compared with CD19 negative (CD19(-)) cases; CD20(+) cases had lower co-expression of CD13 than CD20(-) cases (31.6% vs.67.1%, P = 0.000) and no significant prognostic indications for CD20(+) was observed; CD22(+) cases had higher relapse rate at 12-month than CD22(-) cases (93.9% vs.57.1%, P = 0.041) in B-ALL patients; CD33(+) cases had higher incidence of Ph(+) than CD33(-) cases (43.5% vs.19.4%, P = 0.007) and significantly correlated with Ph(+) (r = 0.261, P = 0.006). It is concluded that elucidation of the characteristics of the target antigens (CD19, CD20, CD22, CD33) used for antibody-based immunotherapy will help hematologists making the correct decision whether and when to use these antibody-based target therapies.
Adolescent ; Adult ; Aged ; Antigens, CD19 ; immunology ; Antigens, CD20 ; immunology ; Child ; Female ; Humans ; Immunophenotyping ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; immunology ; Sialic Acid Binding Ig-like Lectin 2 ; immunology ; Sialic Acid Binding Ig-like Lectin 3 ; immunology ; Young Adult

OBJECTIVETo study the clinicopathologic features, immunohistochemical findings, differential diagnosis and prognosis of type II enteropathy-associated T-cell lymphoma (EATL).

METHODSFourteen cases of type II EATL encountered in Department of Pathology, Nanjing General Hospital were retrospectively reviewed. The clinical data, histologic features, immunohistochemical findings and follow-up information were analyzed, with literature review.

RESULTSThere were altogether 12 males and 2 females. The median age of patient was 49 years. The sites of involvement included jejunum (10 cases) and ileum/colon (4 cases). The patients often presented with an abdominal mass, abdominal pain, diarrhea and constitutional symptoms such as fever, night sweating and cachexia. There was no clinical evidence of gluten-sensitive enteropathy. Histologically, the lymphoma cells showed full-thickness infiltration of the intestinal wall. They contained round hyperchromatic nuclei and pale cytoplasm. The stroma was minimally inflamed, with or without associated coagulative necrosis. A remarkable finding was the presence of villous atrophy, cryptal hyperplasia and intraepithelial lymphocytosis. Immunohistochemical study showed that the tumor cells expressed CD3, CD43 and CD8 (14/14). Some of them were also positive for CD56 (11/14) and CD30 (2/14). The staining for CD4, CD20, CD79a and myeloperoxidase was negative. A high proliferation index was demonstrated by Ki-67 immunostain. In-situ hybridization for EBER was negative. Follow-up data were available in 9 cases. The duration of follow-up ranged from 6 months to 36 months. Seven patients died within 14 months.

CONCLUSIONSEATL is a rare type of lymphoma with intestinal involvement. Associated enteropathy is not demonstrated, in contrast to cases encountered in Nordic countries. A correct diagnosis requires evaluation of clinical manifestations, pathologic features and ancillary study results.

Adolescent ; Adult ; Aged ; CD3 Complex ; metabolism ; CD8 Antigens ; metabolism ; Diagnosis, Differential ; Enteropathy-Associated T-Cell Lymphoma ; genetics ; immunology ; pathology ; surgery ; Female ; Follow-Up Studies ; Gene Rearrangement, T-Lymphocyte ; Humans ; Ileal Neoplasms ; genetics ; immunology ; pathology ; surgery ; Jejunal Neoplasms ; genetics ; immunology ; pathology ; surgery ; Leukosialin ; metabolism ; Lymphoma, B-Cell, Marginal Zone ; metabolism ; pathology ; Lymphoma, Extranodal NK-T-Cell ; metabolism ; pathology ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

OBJECTIVETo study the immunophenotype and chromosome karyotype characteristics of leukemic stem cells (LSC) in Uyghur leukemia pediatric patients.

METHODSThe morphological features of LSC in culture in vitro was observed by flow cytometry. The immunophenotype was assessed by detective flow cytometry. The chromosome karyotype was analyzed by R-banding technique.

RESULTSThe LSC showed suspended floating colonies growing in the culture medium, and grew well and proliferated constantly in culture over 8 months. Among the 13 children with AML, there were 10 CD34(+)CD38(-)CD123(+) and CD33(+) cases, 10 CD44(+) cases, 10 CD96(+) cases, and 5 CD90(+) cases. Among the 13 children with B-ALL, there were 6 CD34(+)CD20(-)CD19(+) cases, 7 CD9(+) cases, and 5 CD123(+) cases. Among the 9 children with acute T lymphoblastic leukemia (T-ALL), there were 5 CD34(+)CD7(-) and CD90(+) cases, and 4 CD123(+) cases. Among the 13 cases of AML, 5 cases showed chromosome translocation t(15;17), one case chromosome translocation t(8;21), and 7 cases showed no chromosome karyotype abnormality. Among the 22 ALL cases, there were chromosome translocation t(12;21) in 1 case, t(9;22) in 3 case, hyperdiploid in 2 cases, and 16 cases without karyotype abnormalities. Twenty-nine children received induction remission therapy. Among them, 12 died, including 9 CD96(-)positive cases and 3 CD96(-)negative cases, with a statistically significant difference (P < 0.05).

CONCLUSIONSThe LSC of Uyghur leukemia pediatric patients in Xinjiang express CD9 and CD19 in ALL, and express CD123 and CD90 simultaneously in ALL and AML. The expression of CD96 is one of factors of poor prognosis.

Adolescent ; Antigens, CD ; metabolism ; Antigens, CD19 ; metabolism ; Child ; China ; ethnology ; Diploidy ; Humans ; Immunophenotyping ; Interleukin-3 Receptor alpha Subunit ; metabolism ; Karyotyping ; Leukemia, Myeloid, Acute ; drug therapy ; genetics ; immunology ; pathology ; Neoplastic Stem Cells ; immunology ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; genetics ; immunology ; pathology ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; genetics ; immunology ; pathology ; Remission Induction ; Tetraspanin-29 ; metabolism ; Thy-1 Antigens ; metabolism ; Translocation, Genetic

OBJECTIVETo perform immunophenotyping and molecular genetic analysis for diffuse large B-cell lymphoma (DLBCL), and to explore their correlation and implication for prognosis.

METHODSImmunohistochemical streptavidin peroxidase (SP) method was used to determine the expression of CD10, BCL6 and MUM1 in 59 cases of DLBCL. A Hans algorithm was used to classify DLBCL into germinal center B-cell (GCB) and non-GCB subtypes. Interphase fluorescence in situ hybridization (FISH) assay was performed on paraffin-embedded lymphoma tissue sections to detect translocations and amplifications of BCL6, BCL2 and MYC genes with dual-color break-apart BCL6 probe, dual-color dual-fusion IgH/ BCL2 probe and dual-color break-apart MYC probe, respectively.

RESULTSIn the 59 cases of DLBCL, 28.8% (17/59) belonged to GCB subtype, and 71.2% (42/59) belonged to non-GCB subtype. The incidences of BCL6, BCL2 and MYC gene translocations were 24.1% (14/58), 1.7% (1/59) and 5.3% (3/57), respectively. The incidences of BCL6, BCL2 and MYC gene amplifications were 17.2% (10/58), 22.0% (13/59) and 21.1% (12/57), respectively. BCL6 amplification was not correlated with BCL6 translocation (P=0.424), but was correlated with amplifications of BCL2 and MYC (C=0.405 and 0.403, respectively, P <0.01). The incidence of BCL6 translocation in GCB type was higher than that in non-GCB type, and amplifications of BCL6, BCL2 or MYC were more frequently encountered in non-GCB type, though no statistical significance was detected (P=0.089 and 0.106, respectively). By univariate analysis, immunophenotyping and international prognostic index (IPI) exerted a significant effect on overall survival (OS) (P=0.047 and 0.001, respectively), but to which BCL6 translocation and amplification of the 3 genes were not related (P=0.150 and 0.444, respectively). By multivariate analysis, IPI score was the only independent prognostic factor for OS (RR =3.843, P=0.017).

CONCLUSIONThe GCB subtype of DLBCL is less common in the patient cohort. Common genetic aberrations have included BCL6 translocation and BCL6, BCL2 and MYC amplifications. Amplification of the 3 genes is strongly correlated with each other, and the incidence of BCL2 translocation is low. Immunophenotyping only has minor significance for the prognosis. Genetic aberrations cannot predict the clinical outcome of DLBCL.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; DNA-Binding Proteins ; genetics ; Female ; Genes, bcl-2 ; Genes, myc ; Humans ; Immunophenotyping ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse ; genetics ; immunology ; Male ; Middle Aged ; Proto-Oncogene Proteins c-bcl-6