Graft-versus-host disease (GVHD) is a life threatening complication that may occur following allogenic bone marrow transplantation (BMT) in the patients with aplastic anemia, leukemia or genetic immunodeficiency. It has been known that GVHD occurs approximately 70% of recipients of BMT in western countries but no definite incidence has been reported in Korea. In our St. Mary's Hospital, GVHD occurs in about 30% of BMT recipients. Histopathologically the acute phase skin shows diffuse lymphocytic infiltrates in the upper dermis with extensive exocytosis. Scattered throughout the epidermis are many degenerated keratinocytes, which are often associated with one or more satellite lymphocytes (satellite cell necrosis). In the chronic phase, acanthosis, eosinophilic keratinocytes resembling colloid bodies and mononuclear cell infiltrates in the upper dermis are noted. We reviewed 5 cases of acute GVHD and 6 cases of chronic GVHD. All patients received allogenic BMT from Jan. 1, 1992 to July 1, 1993. Ten patients were male and one was female. The mean age was 34 (20-70). The pathologic diagnosis was 3 cases of CML, 2 of ALL, 2 of AML (FAB M2), 2 of aplastic anemia, 1 of CLL and 1 of AML (FAB M5). The interval from BMT to GVHD varied from 14 days to 4 years (median 220 days). The skin and GI tract were involved in all eleven cases. Ten cases were histologically proven by skin biopsies, and two cases by salivary gland and colonic biopsies, respectively. The histological findings of the skin, salivary gland and colonic biopsieds were described. Immunohistochemical stain of the skin was done using CD4, CD8, HLA DR and Leu 7 antibodies.
Adult ; Aged ; Antigens, CD4/analysis ; Antigens, CD8/analysis ; Biopsy ; Female ; Graft vs Host Disease/immunology/*pathology ; HLA-DR Antigens/analysis ; Human ; Immunohistochemistry ; Male ; Middle Age

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

The aim of this study was to explore effect of CD8+CD28- T-lymphocyte in the inhibition of mesenchymal stem cells (MSC) on T-lymphocyte proliferation. T cells were harvested by using nylon column and CD8+ T cells were sorted by magnetic beads; the T-lymphocyte proliferation in the presence of PHA was evaluated by MTT; the proportion of CD8+CD28- T cells was assayed by fluorescence-activated cell sorter (FACS). The results showed that MSC inhibited T-lymphocyte proliferation and the inhibitory effect depended on the amount of MSC; the data of FACS indicated that in the CD8+ T cells co-cultured with MSC, CD8+CD28- T cells were up-regulated significantly, compared with the non-treated CD8+ T cells. In conclusion, MSC perform their immunosuppressive function by up-regulation of CD8+CD28- T cells.
Bone Marrow Cells ; physiology ; CD28 Antigens ; analysis ; CD8 Antigens ; analysis ; Humans ; Lymphocyte Activation ; Mesenchymal Stromal Cells ; physiology ; T-Lymphocyte Subsets ; immunology ; Up-Regulation

OBJECTIVETo investigate the relationship between subsets of lymphocytes and between its activated status and the clinical manifestations in patients with PNH, and to unfold immunological mechanism in the pathogenesis of PNH.

METHODSThe peripheral blood mononuclear cells (PBMNC) from 18 PNH patients and 20 controls were separated into two subpopulations using anti-CD(59) monoclonal antibody combined with goat-anti-mouse IgG immunomagnetic beads. CD(3)(+), CD(4)(+) and CD(8)(+) lymphocyte subsets were detected by flow cytometry. In 6 newly diagnosed patients, phenotypes associated with T cell activation such as CD(28)(+)/CD(4)(+) or CD(8)(+) cells, CD(8)(+) CD(38)(+) cells, and HLA-DR(+)/CD(4)(+) or CD(8)(+), and NK (CD(3)(-) CD(16)(+)) cells were detected in the peripheral blood.

RESULTPatients with PNH showed significantly increased CD(3)(+) CD(8)(+)/CD(3)(+) CD(4)(+) ratio as compared with controls (1.22 +/- 0.51 vs 0.86 +/- 0.27, P < 0.05), and the CD(3)(+) CD(8)(+)/CD(3)(+) CD(4)(+) ratio in CD(59)(-) PBMC was higher than that in CD(59)(+) PBMC (2.31 +/- 1.56 vs 0.62 +/- 0.27, P < 0.05). The ratios of CD(4)(+) CD(28)(+)/CD(4)(+) markedly decreased and CD(8)(+)HLA-DR(+)/CD(8)(+) increased.

CONCLUSIONPatients with PNH appear to have abnormalities in their lymphocytes. Increased ratios of CD(3)(+) CD(8)(+)/CD(3)(+) CD(4)(+) and HLA-DR(+) CD(8)(+)/CD(8)(+) lymphocytes as well as declined ratio of CD(4)(+) CD(28)(+)/CD(4)(+) lymphocytes might be involved in the pathogenesis of PNH.

ADP-ribosyl Cyclase ; analysis ; ADP-ribosyl Cyclase 1 ; Adult ; Aged ; Antigens, CD ; analysis ; CD28 Antigens ; analysis ; CD4 Antigens ; analysis ; CD4-Positive T-Lymphocytes ; cytology ; immunology ; CD8 Antigens ; analysis ; CD8-Positive T-Lymphocytes ; cytology ; immunology ; Female ; Hemoglobinuria, Paroxysmal ; blood ; immunology ; Humans ; Killer Cells, Natural ; cytology ; immunology ; Lymphocyte Subsets ; cytology ; immunology ; Male ; Membrane Glycoproteins ; Middle Aged ; Receptors, IgG ; analysis

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

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

Dendritic cells have been known as a member of strong innate immune cells against infectious organelles. In this study, we evaluated the cytokine expression of splenic dendritic cells in chronic mouse toxoplasmosis by tissue cyst-forming Me49 strain and demonstrated the distribution of lymphoid dendritic cells by fluorescence-activated cell sorter (FACS). Pro-inflammatory cytokines, such as IL-1alpha, IL-1beta, IL-6, and IL-10 increased rapidly at week 1 post-infection (PI) and peaked at week 3 PI. Serum IL-10 level followed the similar patterns. FACS analysis showed that the number of CD8alpha+/CD11c+ splenic dendritic cells increased at week 1 and peaked at week 3 PI. In conclusion, mouse splenic dendritic cells showed early and rapid cytokine changes and may have important protective roles in early phases of murine toxoplasmosis.
Animals ; Antigens, CD11c/analysis ; Antigens, CD8/analysis ; Cytokines/*blood/*secretion ; Dendritic Cells/chemistry/*immunology ; Disease Models, Animal ; Flow Cytometry ; Mice ; Mice, Inbred BALB C ; Rodent Diseases/immunology ; Spleen/*immunology ; Time Factors ; Toxoplasmosis, Animal/*immunology

OBJECTIVETo study the activating effect of protein transduction domain (PTD) mediated BCR/ABL protein on T cells from CML patients.

METHODSThe plasmid containing PTD and b3a2 bcr/abl of CML was constructed by genetic engineering and expressed in E. coli. The peripheral blood mononuclear cells from CML patients were stimulated in vitro with purified PTD-BCR/ABL protein and the expression of the early activation antigen CD(69) on CD(8)(+) and CD(4)(+) T cells was detected by flow cytometry (FCM).

RESULTSThe optimal concentration of PTD-BCR/ABL protein for activating CD(8)(+) T cells in vitro was 100 micro g/ml, CD(69) expression peaked in three days stimulation. CD(8)(+) T cells were activated in 10 of 15 CML patients, the expression rate of CD(69) was (15.01 +/- 3.75)%. CD(4)(+) T cells were activated in 4 of 15 patients, the expression rate of CD(69) was (10.32 +/- 3.08)%. Both CD(8)(+) and CD(4)(+) T cells were activated simultaneously in 3 of them. However, neither CD(4)(+) nor CD(8)(+) T cells was activated by stimulation with BCR/ABL protein in all 15 specimens, the expression rate of CD(69) on CD(8)(+) and CD(4)(+) T cells was (1.36 +/- 0.31)% and (1.41 +/- 0.43)%, respectively. There was no difference compared with that of PBS control group (P > 0.05).

CONCLUSIONBy using a PTD-mediated antigen delivering system, exogenous BCR/ABL protein can be delivered into APC, processed and presented onto surface of APC to activate Ag-specific CD(8)(+) and CD(4)(+) T cells in vitro.

Adolescent ; Adult ; Aged ; Amino Acid Sequence ; Antigens, CD ; analysis ; Antigens, Differentiation, T-Lymphocyte ; analysis ; CD4 Antigens ; analysis ; CD8 Antigens ; analysis ; Dose-Response Relationship, Drug ; Female ; Flow Cytometry ; Fusion Proteins, bcr-abl ; genetics ; metabolism ; Gene Products, tat ; genetics ; metabolism ; Humans ; Lectins, C-Type ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; blood ; Male ; Middle Aged ; Recombinant Fusion Proteins ; immunology ; metabolism ; pharmacology ; T-Lymphocytes ; drug effects ; immunology