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

Post-transplantation lymphoproliferative disorders (PTLDs) are a heterogeneous group of diseases that represent serious complications following immunosuppressive therapy for solid organ or hematopoietic-cell recipients. In contrast to B-cell PTLD, T-cell PTLD is less frequent and is not usually associated with Epstein Barr Virus infection. Moreover, to our knowledge, isolated T-cell PTLD involving the breast is extremely rare and this condition has never been reported previously in the literature. Herein, we report a rare case of isolated T-cell PTLD of the breast that occurred after a patient had been treated for allogeneic peripheral blood stem cell transplantation due to acute myeloblastic leukemia.
Allografts ; Axilla ; Breast Neoplasms/diagnosis/*etiology/immunology ; Diagnosis, Differential ; Fatal Outcome ; Female ; Humans ; Leukemia, Myeloid, Acute/surgery ; Lymph Nodes/pathology ; Lymphoma, T-Cell, Peripheral/*etiology/pathology/ultrasonography ; Peripheral Blood Stem Cell Transplantation/*adverse effects ; T-Lymphocytes/immunology/pathology ; Transplantation, Homologous ; Ultrasonography, Mammary/*methods ; Young Adult

This study was aimed to summarize and analyze the clinical features and biological characteristics of adult acute T-lymphoblastic leukemia (T-ALL), and compare the efficacy of chemotherapy and transplantation in order to explore the factors influencing the long term survival and prognosis. Twenty-two T-ALL patients, all of whom were initially diagnosed according to MICM classification criteria from May 2000 to May 2010, were enrolled in this study. All patients received VDCLP regimen as the induction chemotherapy. In consolidation stage, some of the patients received allogeneic hematopoietic stem cell transplantation (allo-HSCT) and the others underwent intensive chemotherapy. The clinical and laboratory parameters were summarized and the contribution to survival and efficacy was analyzed by using χ(2) test, Kaplan-Meier method, Cox regression analysis and log-rank test with the aid of SPSS13.0 software. The results showed that: (1) The median age of all 22 patients was 23.5 years (16 - 63 years). 15 patients with splenomegaly had much shorter event-free survival (EFS) period (P = 0.014) and overall survival (OS) period (P = 0.013). The median white blood cell (WBC) count was 148.82 (5.51-546.0) × 10(9)/L. 15 cases out of them had leucocytosis (WBC ≥ 80 × 10(9)/L), whose EFS period (P = 0.021) and OS time (P = 0.050) were reduced significantly. The similar condition was observed in 6 patients whose blood platelet (Plt) count was no more than 30 × 10(9)/L (P = 0.033 for EFS and P = 0.035 for OS, respectively); (2) Immunophenotypic analysis showed that from 22 cases 2 cases were of pro-T, 14 cases of pre-T, 3 cases of cortical-T and 3 cases of medullary-T. Supposing pro-T and pre-T as earlier period immunophenotype, cortical-T and medullary-T as advanced stage immunophenotype, there were significant differences between earlier period and advanced stage patients in terms of EFS and OS (P = 0.035 for EFS and P = 0.028 for OS, respectively); (3) Chromosome karyotype was analyzed in 19 cases at diagnosis, and among them 12 cases had normal karyotypes while abnormal karyotypes were observed in 7 cases. Correlation analysis showed that there were no significant differences between these two groups in time of EFS and OS; (4) The overall complete remission (CR) rate was 72.7 after the induction chemotherapy. The median CR period was 18.0 months. The EFS and OS rate were 57.9 and 67.1 for 1-year, and 23.0 EFS rate and 22.0 OS rate for 3-years, respectively. Six patients received allo-HSCT and the average EFS time and OS time were both 57.8 months, which were significantly longer than those of the intensive chemotherapy group (P = 0.001 and P = 0.002 for EFS and OS, respectively); (5) Cox regression analysis proved that allo-HSCT treatment was the independent favorable prognostic factor. It is concluded that higher CR rate can be achieved by using intensive induction chemotherapy in adult T-ALL, but the long term survival seems poor by chemotherapy only in consolidation treatment stage. Allo-HSCT is the optimal choice to improve the prognosis and the outcome.
Adolescent ; Adult ; Female ; Hematopoietic Stem Cell Transplantation ; Humans ; Induction Chemotherapy ; Leukemia-Lymphoma, Adult T-Cell ; diagnosis ; immunology ; therapy ; Male ; Middle Aged ; Prognosis ; Remission Induction ; Young Adult

The objective of this study was to investigate the immunophenotype of T-lineage acute lymphoid leukemia (T-ALL) and to find valuable immunologic markers in T-ALL diagnosis and therapy. Four-color multiparametric flow cytometry(FCM) with CD45/SSC gating was used for immunophenotyping of 95 patients with newly diagnosed T-ALL. The results demonstrated that T-ALL occurred more frequently in males younger than 30 years of age and was usually accompanied by a high WBC count and tumor mass at diagnosis. Univariate analysis showed an influence on achievement of CR1 for age (< 30 years) but not for WBC count and tumor mass. According to WHO (2008) classification of tumors of haematopoietic and lymphoid tissues, 87 patients with confirmed subtype included 27 cases of Pro-T-ALL (31.0%), 31 cases of Pre-T-ALL (35.6%), 23 cases of cortical-T-ALL (26.4%), 6 cases of medullary-T-ALL (6.9%). CD34 expression in Pro-T-ALL was significantly higher than that of Pre-T-ALL (p = 0.001). After the first chemotherapy, the complete remission rate in Pro-T-ALL was statistically lower than that of Pre-T-ALL. Besides, the complete remission rate of immature T-ALL (including Pro-T-ALL and Pre-T-ALL) was also significantly lower than that in mature T-ALL (including cortical-T-ALL and medullary-T-ALL). Myeloid antigen (CD13, CD33) expression was associated with T-ALL subtype and treatment effect. While 66.7% of CD13(+) patients belonged to Pre-T-ALL, most (60.0%) of CD33(+) patients were classified into Pro-T-ALL; CD13 expression had no effect on CR1 rate whereas CD33(+) patients had worse treatment effect compared with CD33(-) groups (p = 0.001). Notably, the expression of CD117 reached up to 26.7% and the positive cases were primarily distributed in pro-T-TAll and pre-T-ALL. It is found that CD117 expression in CD34(-) group was homogeneous and CD117 expression level was less than 10% in 73.2% patients, but CD117 expression level in CD34(+) group was not homogenous, in which group the CD117 expression levels < 10%, 10% - 20% and > 20% were 44.2%, 17.3% and 38.5% respectively. As compared with CD34(-) group, the proportion of patients with CD117 expression levels < 10%, > 20% in CD34(+) group was higher, and there was significant difference between these 2 group. It is concluded that immunophenotype has great value in T-ALL diagnosis, classification as well as treatment. Flow cytometry provides access to find valuable immunologic markers for T-ALL biological research.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; CD13 Antigens ; metabolism ; Child ; Child, Preschool ; Female ; Humans ; Immunophenotyping ; Infant ; Male ; Middle Aged ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; classification ; immunology ; therapy ; Proto-Oncogene Proteins c-kit ; metabolism ; Sialic Acid Binding Ig-like Lectin 3 ; metabolism ; Young Adult

BACKGROUND AND OBJECTIVEPrecursor T lymphoblastic lymphoma (T-LBL) is a highly aggressive lymphoma. Myeloid antigen expression was found in some of the patients, and its clinical significance is worth studying. This study was to compare the clinical features, short-term efficacy and survival of T-LBL patients with or without myeloid antigen expression so as to evaluate its prognostic significance.

METHODSForty-five T-LBL patients, with a median age of 14 years, were treated at Sun Yet-sen University Cancer Center between January 2000 and July 2008. These patients were divided into myeloid antigen-positive group (My(+) group) and myeloid antigen-negative group (My(-) group) based on the flow cytometric (FCM) analysis in bone marrow or pleural fluid. Myeloid antigen expression and its correlation with the short-term efficacy and overall survival were assessed in the two groups.

RESULTSThere were 18 patients (40.0%) in the My(+) group and 27 (60.0%) in the My(-) group. The myeloid antigen expression was negatively correlated with the initial level of lactate dehydrogenase (LDH), but not with other clinical features. The remission rate was lower in the My(+) group than in the My(-) group (38.8% vs. 70.3%, P = 0.028). The 2-year overall survival rate was lower in the My(+) group than in the My(-) group (51.9% vs. 78.7%, P = 0.036). By age subgroup analysis, there were no differences in response and survival rate among children and adolescents with or without myeloid antigen expression. But the remission rate and the 2-year overall survival rate were significantly lower in adult patients with myeloid antigen expression than in patients without it. Univariate and multivariate analysis demonstrated that age and myeloid antigen expression were adverse prognostic factors.

CONCLUSIONMyeloid antigen expression is a predictor of a poor response to chemotherapy, and adverse prognostic factor in adult T-LBL, but not in children with T-LBL.

Adolescent ; Adult ; Age Factors ; Aged ; Antigens, CD7 ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Asparaginase ; therapeutic use ; Child ; Cyclin D3 ; metabolism ; Cyclophosphamide ; therapeutic use ; Cytarabine ; therapeutic use ; Daunorubicin ; therapeutic use ; Doxorubicin ; therapeutic use ; Etoposide ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Male ; Mercaptopurine ; therapeutic use ; Methotrexate ; therapeutic use ; Middle Aged ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; immunology ; Prednisone ; therapeutic use ; Proportional Hazards Models ; Remission Induction ; Survival Rate ; Transcription Factors ; metabolism ; Vincristine ; therapeutic use ; Young Adult

The aim of the study was to analyze the naive T cell level of thymic recent output in patients with B-cell malignancies, thereby to evaluate the potential T-cell function. Quantitative analysis of T-cell receptor rearrangement excision circles (TRECs) in DNA of peripheral blood mononuclear cells from 61 cases of B-cell lymphocytic malignancy (including 20 cases of adult B-ALL, 6 case of childhood B-ALL, 4 cases of B-CLL, 17 cases of B-NHL and 14 cases of MM) were preformed by real-time PCR (TaqMan), and TREC-level was detected according to the number of CD3-positive cells. 5 case of ALL-CR and 17 normal individuals were served as controls. The results showed a dramatic reduction of TREC values in all groups of patients. The mean value of TRECs was 0.53 +/- 1.52 copies/1000 PBMNC and 2.01 +/- 3.93 copies/1000 CD3+ cells in adult B-ALL (p = 0.0005, p = 0.0123), 0.11 +/- 0.15 copies/1000 PBMNC and 0.23 +/- 0.27 copies/1000 CD3+ cells in B-CLL (p = 0.0015, p = 0.0381), 0.71 +/- 1.34 copies/1000 PBMNC in B-NHL (p = 0.0017), 0.53 +/- 0.90 copies/1000 PBMNC in MM patients (p = 0.0018), as compared with 3.76 +/- 3.42 copies/1000 PBMNC and 5.87 +/- 4.96 copies/1000 CD3+ cells in normal individuals, the TREC level was significantly decreased in all groups of B-cell lymphocytic malignancy, as well as in ALL-CR group. However, the TREC level in childhood B-ALL was significant higher than those in adult B-ALL group. It is concluded that the function of thymic recent outputting naive T cells in B-cell malignancies significantly decreases, however, the individual difference of thymic output function is obvious. The thymic recent output function can not be recovered during CR phase in patients with B-cell malignancies, so that dynamic analysis of TREC level is necessary.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; B-Lymphocytes ; metabolism ; pathology ; Female ; Gene Rearrangement, T-Lymphocyte ; Humans ; Male ; Middle Aged ; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; immunology ; pathology ; T-Lymphocytes ; immunology ; Thymus Gland ; immunology ; metabolism ; Young Adult

The objective of this study was to investigate the immunophenotypic characteristics of T-cell acute lymphoblastic leukemia (T-ALL). Immunophenotyping was performed in 140 T-ALL patients by flow cytometry using a panel of monoclonal antibodies and CD45/SSC gating. The results showed that the T-lineage-associated antigen expressions were CD7 > CD2 > CD3 > CD5 successively. The positive rate of CD10 was 19.42% in patients. Among 140 cases of T-ALL, 12 (8.57%) was accompanied by B-lineage associated antigen expression. Myeloid antigen expression was identified in 31 out of 136 cases (22.79%). None of them expressed CD14 antigen. The positive rate of CD34 was 31.06%. The positive rate of myeloid antigen expression in CD34(+) T-ALL (36.58%) was significantly higher than that in CD34(-) T-ALL (15.38%) (p < 0.01). The expression of CD3 in child T-ALL was higher than that in adult T-ALL, whereas the expression of CD33 in children was lower than that in adults. It is concluded that immunophenotyping is an important tool for diagnosis of T-ALL. Immunophenotypic characteristics of T-ALL is heterogeneous.
Adolescent ; Adult ; Aged ; Antigens, CD ; metabolism ; Antigens, CD34 ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; CD3 Complex ; metabolism ; Child ; Child, Preschool ; Female ; Humans ; Immunophenotyping ; Infant ; Male ; Middle Aged ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; immunology ; Sialic Acid Binding Ig-like Lectin 3 ; Young Adult

The aim was to investigate the immunophenotypes of NK series leukemia. Immunophenotypes of 297 cases of acute leukemia (AL) were measured by flow cytometry, and these immucopenotypic features were analyzed. The results showed that 43 out of 297 cases of AL (14.5%) were CD56 positive. 6 cases were NK series leukemia and 37 cases were acute myelogenous leukemia with CD56 expressed. One patient has been diagnosed as myeloid/NK cell precursor acute leukemia, two patients were blastic NK cell leukemia, one was supposed to be NK-like T-cell lymphoma/leukemia, while another one was large granular lymphocyte leukemia (LGLL). It is concluded that almost all of CD56 positive leukemia were acute myelogenous leukemia with CD56 expressed. The immunophenotypes of NK series leukemia were antigens from hematopoietic stem cells to T/NK progenitor cells with meyloid antigen positive, and through NK progenitors to mature NK cells. The immunophenotypes of heterogeneous NK leukemia cells are different, that should be carefully distinguished.
Adolescent ; Adult ; Aged ; CD56 Antigen ; metabolism ; Female ; Humans ; Immunophenotyping ; Killer Cells, Natural ; immunology ; Leukemia, Myeloid, Acute ; immunology ; Lymphoma, T-Cell ; immunology ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; immunology

OBJECTIVETo investigate the clinical efficacy of Shenqi Fuzheng Injection (SFI) combined with chemotherapy in treatment of patients with acute leukemia and its effect on the levels of T-lymphocyte subsets (CD4, CD8, CD4/CD8) and serum interferon-gamma(IFN-gamma), interleukin-10 (IL-10) and IL-2.

METHODSSixty-five patients with initial treating acute leukemia were randomly divided into 2 groups, the SFI group (n = 32) treated with SFI plus chemotherapy (CT), the control group (n = 33) treated with CT only. The remission rate, changes of peripheral mature neutrophilic granulocyte (PMNG) count, T-lymphocyte subsets, serum IL-10 and IL-2 before and after treatment were determined.

RESULTSThe remission rate in the two groups showed no obvious difference (P > 0.05). After CT for the 1st, 2nd and 3rd weeks, the PMNG count decreased in both groups, showing significant difference as compared with that before CT (P < 0.01 or P < 0.05). The PMNG count at the end of the 3rd and 4th week of CT remounted to higher than that at 1st and 2nd week, and the increment in the SFI group was significantly higher than that in the control group (P < 0.05). The levels of CD4, CD4 /CD8, IFN-gamma and IL-2 all increased in the two groups after treatment (P < 0.05, P < 0.01), however, that of IL-10 was significantly decreased (P < 0.01). The difference between the two groups in these criteria after treatment was also significant (P < 0.05).

CONCLUSIONSFI can improve and regulate the immune function of the patients with acute leukemia undergoing CT, it could promote bone marrow cells proliferation and enhance the efficacy.

Adolescent ; Adult ; Aged ; Antineoplastic Agents, Phytogenic ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Interleukin-2 ; blood ; Leukemia, Myeloid, Acute ; drug therapy ; immunology ; Male ; Middle Aged ; Phytotherapy ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; immunology ; T-Lymphocyte Subsets ; immunology

OBJECTIVETo analyse the correlation between the cellular immunity and TCM Syndrome typing of patients with minimal residual leukemia (MRI).

METHODSThe analysis was performed by detecting cellular immunological parameters in 30 MRI patients, 55 healthy persons and 36 patients with acute leukemia (AL) using three fluoresceine-conjugated monoclonal antibodies and flow cytometer.

RESULTSAs compared with those in the healthy persons, universal reductions of various parameters were shown in the MRI patients, including WBC count, absolute value of total T-lymphocytes, T-helper lymphocyte (P < 0.05) and total lymphocytes; the percentage and absolute value of NK cells (P < 0.01); and the percentages of total T-lymphocytes, CD4+ CD29+, T-suppressor cells and T-memory cells (P < 0.05 or P < 0.01), but without any rising of absolute value. As compared with those in the patients with AL, parameters were similar in the two groups with insignificant difference. The disturbances, including the lowering on ratio of T-helper/T-suppressor lymphocytes, in MRI patients of Qi-blood insufficiency type was the severest, that in the patients of Qi-Yin deficiency type was the mildest, and that in patients of Yin-deficiency with excess Fire type located between them.

CONCLUSIONThe immune function of MRI patients is low, belonging to the TCM Syndrome of vital energy deficiency with evil-lingering. Since the degree of cellular immune disturbance is different in various TCM Syndrome types, therefore, they should be treated with different dosages of different drugs.

Adolescent ; Adult ; Aged ; CD4-CD8 Ratio ; Child ; Diagnosis, Differential ; Female ; Humans ; Immunity, Cellular ; Killer Cells, Natural ; immunology ; Leukemia, Myeloid, Acute ; immunology ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Neoplasm, Residual ; immunology ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; immunology ; T-Lymphocyte Subsets ; immunology ; Yang Deficiency ; immunology ; Yin Deficiency ; immunology