Fine-needle aspiration (FNA) of lymph nodes has been regarded as a useful method in the diagnosis of lymphadenopathy. However, this procedure has been shown to be of limited value in the diagnosis of low or intermediate grade malignant lymphomas in some studies. Immunophenotyping is an essential adjunct to cytomorphology for the diagnosis of lymphoma by FNA. Immunophenotyping using flow cytometry (FCM) is rapid, objective and reliable. Using FCM, multiparametric analysis of 33 FNA materials from lymph nodes was performed and profiles of surface markers of lymphoid cells were assessed. In reactive hyperplasia, patterns of cell surface markers were quite variable, but disclosed polyclonality. Most of the B-cell lymphomas showed immunophenotypes for B-cell lineages with their kappa: lambda or lambda: kappa ratio being over 3:1. In T-cell lymphomas, T-cell surface markers were predominantly expressed as well. In conclusion, our results suggest that immunophenotyping of lymph node aspirates is a valuable diagnostic adjunct for lymphoproliferative disorders, particularly in B-cell lymphomas because immunophenotyping can be easily and adequately performed by FCM.
Antigens, CD19/analysis ; Antigens, CD20/analysis ; Antigens, CD3/analysis ; Antigens, CD4/analysis ; Antigens, CD5/analysis ; Antigens, CD7/analysis ; Antigens, CD8/analysis ; B-Lymphocytes/immunology ; B-Lymphocytes/chemistry ; Biopsy, Needle ; Flow Cytometry/methods* ; Hodgkin Disease/pathology ; Human ; Immunophenotyping ; Lymph Nodes/pathology ; Lymph Nodes/chemistry ; Lymphatic Diseases/pathology* ; Lymphatic Metastasis/pathology ; Lymphoma, B-Cell/pathology* ; Lymphoma, Non-Hodgkin/pathology ; T-Lymphocytes/immunology ; T-Lymphocytes/chemistryt

OBJECTIVETo determine the effect of early enteral nutrition (EN) on immune function of the patients after operation for severe abdominal trauma.

METHODSFourty patients who underwent operation for severe abdominal trauma were randomly divided into two groups, and received an early enteral nutrition (EN group, n=20) through jejunal nutritional tube from postoperative day 1, or parental nutrition (PN group, n=20) for 7 days. C3, IgA, IgM, IgG and CD3+, CD4+, CD8+, CD4+/CD8+ of the two groups patients were detected on the day before operation and the postoperative day 1 and 8. The infection complications were compared.

RESULTSAfter 7 days, the levels of C3+, IgA, IgG, CD3+, CD4+, CD8+, and CD4+/CD8+ in EN group increased significantly compared with those in PN group (P< 0.05). The incidence of infection was 10% in EN group, while 30% in PN group (P< 0.05).

CONCLUSIONEarly enteral nutrition can improve immune function and decrease postoperative infection after operation for severe abdominal trauma.

Abdominal Injuries ; immunology ; surgery ; Adolescent ; Adult ; CD3 Complex ; analysis ; CD4 Antigens ; analysis ; CD4-CD8 Ratio ; CD8 Antigens ; analysis ; Complement C3 ; analysis ; Enteral Nutrition ; Female ; Humans ; Immunoglobulin A ; analysis ; Immunoglobulin G ; analysis ; Male ; Postoperative Period ; Young Adult

Although active inflammation may be deleterious and indicate immunologic activation in chronically rejected grafts, the underlying mechanism of tissue destruction has been little studied. Twenty-four cases of chronic rejection (CR) with or without acute rejection (AR) were stained with antibodies against CD3, CD8, CD68, granzyme B and TIA-1, and the number of positive cells were counted. Eleven cases of AR served as controls. The number of CD3 and CD8 positive cells increased in the acute on CR group compared to the CR group. About a half of CD3 positive T cells were CD8 positive in both groups, however, the proportion of TIA-1 or granzyme B positive cells was higher in the acute on CR group. The numbers of CD3, CD68, granzyme B and TIA-1 positive cells were higher in the AR group than the acute on CR group, however, no significant difference was found between the two groups. Serum creatinine level and proteinuria at the time of biopsy and the percentages of late onset AR and graft failure rate were higher in the acute on CR group than the CR group. Summarizing, these results suggest that infiltration of activated T cells containing cytotoxic granules plays a role in graft destruction in acute on CR.
Adult ; Antigens, CD3/analysis ; Antigens, CD8/analysis ; Female ; Follow-Up Studies ; *Graft Rejection ; Human ; Immunohistochemistry ; *Kidney Transplantation ; Male ; Membrane Proteins/*analysis ; RNA-Binding Proteins/*analysis ; Serine Endopeptidases/*metabolism ; Transplantation, Homologous

The purpose was to study the effect of mesenchymal stem cell (MSC) on immune function, and to explore the new strategy to prevent graft versus host disease (GVHD) and host versus graft reaction (HVGR) in allogeneic bone marrow transplantation. MSCs from human bone marrow cells were isolated and cultured. The purity of MSCs were identified with the spindle-fibroblastic morphological characterization by microphotography, and the phenotypes were tested by flow cytometry. MSCs were planted in 6-well plates (8 x 10(4)/well for group A, 4 x 10(4)/well for group B and 2 x 10(4)/well for group C), and cocultured for 7 days with T cell isolated from peripheral blood. Peripheral blood T cells non-cocultured with MSC acted as the control group. CD3, CD4, CD8, and CD25 expressed on T cells were analyzed by flow cytometry after coculture with MSCs for 0, 24, 72 hours and 7 days respectively. The results showed that CD4(+)CD25(+) T cells and CD8(+) T cells of allogeneic T lymphocytes cocultured with bone marrow MSCs (group A and group B) increased obviously as compared with control group (T lymphocytes uncocultured with MSCs), and there were no difference between groups A and B. CD3, CD4, CD8 and CD25 expressed on T cells had no significant difference between experimental groups and control group. The result suggested that CD4(+)CD25(+)-regulatory T cells and CD8(+) T cells were increased apparently when cocultured with allogeneic MSCs. It is concluded that MSCs pretreatment may be useful in the prevention of GVHD and HVGR in allogeneic bone marrow transplantation and provides a new strategy to induce transplantation tolerance.
Bone Marrow Cells ; cytology ; immunology ; CD3 Complex ; analysis ; CD4 Antigens ; analysis ; CD8 Antigens ; analysis ; Cell Communication ; immunology ; Cells, Cultured ; Coculture Techniques ; Flow Cytometry ; Humans ; Immunophenotyping ; Interleukin-2 Receptor alpha Subunit ; analysis ; Mesenchymal Stromal Cells ; cytology ; immunology ; T-Lymphocytes ; cytology ; immunology

The present study was conducted in order to investigate the immunologic alterations alongside the numerical changes in peripheral blood lymphocytes(PBL) and their subsets in stomach cancer patients. Lymphocyte surface markers were determined in 85 stomach cancer patients and 49 controls by indirect immunofluorescence technique using monoclonal antibodies. Monoclonal antibodies used were Leu 2a(CD8, suppressor/cytotoxic T cells), Leu 3a(CD4, inducer/helper T cells), Leu 4(CD3, pan T reagent), Leu 11(CD16, natural killer cells) and Leu 12(CD19, B cells). The numbers of PBL, CD3+, CD4+, CD8+, CD16+ and CD19+ cells significantly decreased and the CD4: CD8 value increased in 85 patients with stomach cancer compared to those in controls(p < 0.01). In stage I(n = 17), neither PBL, their subsets nor the CD4: CD8 value were significantly different from those of the controls. In stage II(n = 17), the numbers of PBL, CD3+, CD4+ and CD8+ cells decreased(p < 0.01). In stage III(n = 24) and IV(n = 27), PBL and all subsets measured decreased(p < 0.01). The CD4: CD8 value showed significant increases in stages III and IV(p < 0.01), because the CD8+ cells decreased to a greater extent than did the CD4+ cells. The results demonstrating that the lymphocyte subsets are depressed differentially with the stage suggest that host immunity is impaired with the progression of stomach cancer.
Adult ; Age Distribution ; Aged ; Antigens, CD19/analysis ; Antigens, CD3/analysis ; CD4-Positive T-Lymphocytes/cytology ; CD8-Positive T-Lymphocytes/cytology ; Female ; Human ; Lymphocyte Count ; Lymphocyte Subsets/chemistry/*immunology ; Male ; Middle Age ; Receptors, IgG/analysis ; Sex Distribution ; Stomach Neoplasms/blood/*immunologym

Establishing Epstein-Barr virus(EBV)-specific cytolytic T lymphocytes(EBV-CTLs) from peripheral blood mononuclear cells(PBMCs) for adoptive immunotherapy has been reported in EBV-associated malignancies including Hodgkin's lymphoma and nasopharyngeal carcinoma(NPC). In the current study, we performed ex vivo expansion of tumor-infiltrating lymphocytes(TILs) obtained from NPC biopsy specimens with a rapid expansion protocol using anti-CD3 monoclonal antibody(OKT3), recombinant human interleukin(IL)-2, and irradiated PBMCs from healthy donors to initiate the growth of TILs. Young TIL cultures comprised of more than 90% of CD3+ T cells, a variable percentage of CD3+CD8+ and CD3+CD4+ T cells, and less than 10% of CD3-CD16+ natural killer cells, a similar phenotype of EBV-CTL cultures from PBMCs. Interestingly, TIL cultures secreted high levels of the Th1 cytokines, interferon gamma (IFNγ) and tumor necrosis factor-alpha (TNF-α), and low levels of the Th2 cytokines, IL-4 and IL-10. Moreover, young TILs could recognize autologous EBV-transformed B lymphoblast cell lines, but not autologous EBV-negative blast cells or allogeneic EBV-negative tumor cells. Taken together, these data suggest that ex vivo expansion of TILs from NPC biopsy tissue is an appealing alternative method to establish T cell-based immunotherapy for NPC.
Biopsy ; CD3 Complex ; analysis ; CD4 Antigens ; analysis ; CD8 Antigens ; analysis ; Cells, Cultured ; Herpesvirus 4, Human ; immunology ; Humans ; Immunotherapy, Adoptive ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-2 ; pharmacology ; Interleukin-4 ; metabolism ; Lymphocytes, Tumor-Infiltrating ; immunology ; virology ; Monocytes ; pathology ; Muromonab-CD3 ; pharmacology ; Nasopharyngeal Neoplasms ; immunology ; pathology ; therapy ; virology ; Receptors, IgG ; analysis ; T-Lymphocytes, Cytotoxic ; immunology ; virology ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo study the levels of CD4+CD25+CD127- and CD3+CD4-CD8- regulatory T (Treg) cells in peripheral blood of children with idiopathic thrombocytopenic purpura (ITP).

METHODSThe flow cytometry was used to detect the expression of CD4+CD25+CD127- and CD3+CD4-CD8- Treg cells in peripheral blood of 33 children with ITP and 21 healthy children.

RESULTSThe expression levels of CD4+CD25+CD127-［(2.7±1.7)% vs (4.8±1.6)%; P<0.01］and CD3+CD4-CD8-［(5.2±3.1)% vs (8.1±3.5)%; P<0.01］Treg cells in children with ITP were significantly lower than in the controls.

CONCLUSIONSThe expression levels of CD4+CD25+CD127- and CD3+CD4-CD8- Treg cells decrease in children with ITP, suggesting that CD4+CD25+CD127- and CD3+CD4-CD8- Treg cells might play a role in the pathogenesis of ITP.

Adolescent ; CD3 Complex ; analysis ; CD4 Antigens ; analysis ; Child ; Child, Preschool ; Female ; Flow Cytometry ; Humans ; Infant ; Interleukin-2 Receptor alpha Subunit ; analysis ; Interleukin-7 Receptor alpha Subunit ; analysis ; Male ; Purpura, Thrombocytopenic, Idiopathic ; etiology ; immunology ; T-Lymphocytes, Regulatory ; immunology

Hepatosplenic gammadelta T cell lymphoma represents rare, often aggressive type of malignant peripheral T-cell lymphoma, which is characterized by expressing T-cell-associated markers CD2, CD3 and gammadelta T-cell receptor, and nonactivated cytotoxic cell phenotype (TIA-1+, granzyme B-). The pathological findings of a liver biopsy specimen revealed the diffuse infiltration of lymphocytes in the sinusoids and the aspiration biopsy from spleen revealed the diffuse infiltration of lymphocytes in the red pulp, not shaped to the nodes, often resulted in the misdiagnosis. Recently, by analyzing the immunophenotype and TCR rearrangement from liver, spleen and bone marrow, a case of adult hepatosplenic gammadelta T cell lymphoma was diagnosed. In combination with references, It is belived that immunophenotype and TCR rearrangement are necessary means to diagnosis hepatosplenic gammadelta T cell lymphoma.
Adult ; Antigens, CD20 ; metabolism ; CD2 Antigens ; analysis ; CD3 Complex ; metabolism ; Humans ; Immunohistochemistry ; Ki-1 Antigen ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Lymphoma, T-Cell ; metabolism ; pathology ; Male ; Receptors, Antigen, T-Cell, gamma-delta ; metabolism ; Splenic Neoplasms ; metabolism ; pathology

Adolescent ; CD3 Complex ; blood ; CD4 Antigens ; blood ; CD8 Antigens ; blood ; Child ; Fetal Blood ; immunology ; Humans ; Infant, Newborn ; Interleukin-6 ; blood ; Interleukin-8 ; blood ; Male ; Nitric Oxide ; blood ; T-Lymphocytes ; immunology ; Tumor Necrosis Factor-alpha ; analysis

OBJECTIVETo define the mechanism of acute hepatitis in non-human primates after liver directed gene therapy.

METHODSDifferences in immune response exhibited by 8 rhesus monkeys receiving adenovirus (Ad) or lipofectamine-mediated gene transfer by various routes, the time course, and the nature of the specific immune responses to both adenoviral vectors and transgene products were studied using HE staining (H&E) and immunohistochemical staining.

RESULTSThe monkeys developed mild to moderate acute hepatitis 1 to 3 weeks after intravenous or intrabiliary injection of first generation replication-defective adenoviruses carrying the Escherichia coli lacZ gene. This was accompanied by adenovirus-mediated T-cell proliferation and neutralizing antibodies to the adenovirus. Increased numbers of CD3(+), CD4(+) and CD8(+) T-lymphocytes were detected in the diseased livers, while B-lymphocytes were absent. Hepatocytes demonstrated increased expression of beta 2-microglobulins (beta 2-MG) and HLA-DR antigens in the plasma membranes. The development of acute hepatitis and the accompanying immune abnormalities were delayed in immunosuppressed monkeys until after the discontinuation of immunosuppressive therapy. The monkeys infused with Ad. CMVluc showed more significant and longer durations of hepatitis than the monkeys infused with adenoviruses carrying the lacZ gene. Lipofectamine-mediated gene transfer was inefficient. There was neither lacZ expression nor significant immune response in the liver of monkeys infused with lipofectamine via the portal vein or the common bile duct.

CONCLUSIONImmune response to the hepatocytes in liver directed gene therapy is MHC class I restricted and T-cell mediated. Both adenoviral vectors and foreign genes are related to the liver damage. Mild to moderate hepatic inflammation seen with the E-1 deleted vector is reversible. Immunosuppression regimens may prolong transgene expression and delay the development of acute adenoviral hepatitis.

Acute Disease ; Adenoviridae ; genetics ; Adenoviridae Infections ; genetics ; Animals ; CD3 Complex ; analysis ; CD4 Antigens ; analysis ; CD8 Antigens ; analysis ; DNA, Recombinant ; administration & dosage ; genetics ; Gene Transfer Techniques ; HLA-DR Antigens ; analysis ; Hepatitis, Animal ; genetics ; virology ; Immunohistochemistry ; Liver ; chemistry ; metabolism ; pathology ; Macaca mulatta ; beta 2-Microglobulin ; analysis