Hematopoietic neoplasm coexpressing CD4 and CD56 includes a subset of acute myeloid leukemia with myelomonocytic differentiation, plasmacytoid monocyte tumor, and other immature hematopoietic neoplasms of undefined origin. Herein, we report a CD4+CD56+CD68+ hematopoietic tumor that was thought to be a tumor of plasmacytoid monocytes. This case is unique in the absence of accompanying myelomonocytic leukemia and the faint expression of cCD3 on the tumor cells. The patient was a 22-yr old man presented with multiple lymphadenopathy and an involvement of the bone marrow. Tumor cells were large and monomorphic with an angulated eosinophilic cytoplasm of moderate amount. Nuclei of most tumor cells were eccentric and round with one or two prominent nucleoli. Rough endoplasmic reticulum was prominent in electron microscopic examination. Tumor cells expressed CD4, CD7, CD10, CD45RB, CD56, CD68, and HLA-DR and were negative for CD1a, CD2, sCD3, CD5, CD13, CD14, CD20, CD33, CD34, CD43, CD45RA, TIA-1, S-100, and TdT. cCD3 was not detected in the immunostaining using paraffin tissue, but was faintly expressed in flow cytometry and immunostaining using a touch imprint slide. T-cell receptor gene rearrangement analysis and EBV in situ hybridization showed negative results. Cytochemically, myeloperoxidase, Sudan black B, and alpha naphthyl butyrate esterase were all negative.
Adult ; Antigens, CD/*biosynthesis ; Antigens, CD3/*biosynthesis ; Antigens, CD4/*biosynthesis ; Antigens, CD45/biosynthesis ; Antigens, CD56/*biosynthesis ; Antigens, Differentiation, Myelomonocytic/*biosynthesis ; Bone Marrow Cells/pathology ; Cell Nucleus/pathology ; Eosinophils/metabolism ; Flow Cytometry ; Gene Rearrangement ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Leukemia, Lymphocytic, Chronic, B-Cell/diagnosis/*metabolism ; Lymph Nodes/pathology ; Male ; Microscopy, Electron ; Monocytes/*metabolism ; Receptors, Antigen, T-Cell/metabolism

In order to study how to activate T cells and their immunological characteristics, the anti-CD3 and anti-CD28 McAbs were used to stimulate PBMNC, then their related immunological changes, such as lymphocyte transformation function, the percentage of CD8(+)CD25(+) cells and TGF-beta expression were deleted by lymphocyte transformation assay, flow cytometry and RT-PCR respectively. The results showed that in costimulation with anti-CD28 antibody stimulation, the activity of anti-CD3 antibody was significantly enhanced, the ratio of CD8(+)CD25(+) cells of T cells was obviously increased, while TGF-beta expression was down-regulated. It was concluded that the anti-CD28 antibody costimulation could provide stimulatory signal II, which make T cells more active, while the expression of TGF-beta significantly down-regulated.
Antibodies, Monoclonal ; pharmacology ; CD28 Antigens ; immunology ; CD3 Complex ; immunology ; Down-Regulation ; Humans ; Leukocytes, Mononuclear ; immunology ; metabolism ; Lymphocyte Activation ; drug effects ; T-Lymphocytes ; immunology ; Transforming Growth Factor beta ; biosynthesis

The purpose of this study was to optimize a fixation procedure for detection of cytoplasmic antigens by flow cytometry(FCM) and to evaluate the effect of intracellular CD3, CD22, CD79a and myeloperoxidase(MPO) in lineage assignment. Four kinds of fixation procedure and three or four color direct immunofluorescence staining were used to permeate cell membrane and label cell surface and intracellular antigens by means of FCM. Results showed that percentage of cytoplasmic antigens positive cells was the highest and cell scatter and fluorescence intensity of CD45 were not changed after using of FACS permeabilization solution. MPO protein was positive in 16/18 acute myeloid leukemia(AML) patients. 4 cases of T cell-acute lymphoblastic leukemia (T-ALL) cases were positive for cytoplasmic CD3(c CD3) but surface CD3 was negative. c CD22 was only detected in 9/13 of B-ALL and cCD79a was positive in 5/5 B-ALL. 18/38 cases of acute leukemia were expressed in more than one lineage marker, 8/21 cases of acute non-lymphocytic leukemia(ANLL) were CD7 positive. 7/17 cases of acute lymphocytic leukemia (ALL) expressed CD13. After further cytoplasmic antigen detection, one was considered to be a T/myeloid biphenotypic leukemia, another one was diagnosed as biclonal or mixed leukemia. The results suggest that intracellular CD3,CD22,CD79a and MPO are lineage specific markers, they are very important for biphenotypic and biclonal/mixed acute leukemia identification
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD ; analysis ; biosynthesis ; Antigens, Differentiation, B-Lymphocyte ; biosynthesis ; CD3 Complex ; biosynthesis ; CD79 Antigens ; Cell Adhesion Molecules ; Child ; Child, Preschool ; Diagnostic Techniques and Procedures ; Female ; Flow Cytometry ; Humans ; Immunophenotyping ; methods ; Lectins ; biosynthesis ; Leukemia ; classification ; pathology ; Male ; Middle Aged ; Peroxidase ; biosynthesis ; Receptors, Antigen, B-Cell ; biosynthesis ; Sialic Acid Binding Ig-like Lectin 2

OBJECTIVETo investigate the effect of mitogen Phorbol 12-myristate 13-Acetate (PMA) on CD3, CD4 and CD8 expression of human T-lymphocytes.

METHODSPeripheral blood mononuclear cells from 37 blood samples stimulated in vitro with PMA at different concentrations (2,5,10,20 and 50 ng/ml for 4 hours) and time (10 ng/ml for 2,4 and 6 hours) were analyzed by 4-color flow cytometry (FCM).

RESULTSUnder different PMA stimulation protocols,significant CD4 down-regulation was observed,which was negatively correlated with intracellular cytokine secretion (r= 0.601,P<.001), except for PMA stimulation at 10 ng/ml for 2-hours which showed no significant intracellular cytokine secretion. The expressions of CD3 and CD8 molecules after PMA activation were not significantly affected as compared with pre-activation. Among CD3 positive T lymphocytes, CD4/CD8 double-negative cells only account for 5.52%.

CONCLUSIONPMA has a significant down-regulation effect on CD4 molecules of Th cells, without altering the CD3 and CD8 expression. For quantitative analysis of Th1/Th2 variation, indirect method such as CD3(+)CD8( ) T cells can be used to define CD4(+) Th cells stimulated by PMA in the future.

CD3 Complex ; blood ; CD4 Antigens ; blood ; CD8 Antigens ; blood ; Child ; Humans ; Interferon-gamma ; biosynthesis ; Interleukin-4 ; biosynthesis ; Lymphocyte Activation ; drug effects ; T-Lymphocytes ; chemistry ; drug effects ; Tetradecanoylphorbol Acetate ; pharmacology ; Th1 Cells ; immunology ; Th2 Cells ; immunology

The synthetic gene with 1640bp encoding for the anti-CD3/anti-CD20 bispecific single-chain antibody was designed and obtained by SOE (splicing by overlap extension) PCR. The cDNA was cloned into Flp-In expression vector pcDNA5/FRT and transfeced into Flp-In CHO cells to generate a stable expression cell line with a capacity for expressing anti-CD3/anti-CD20 bispecific single-chain antibody at 300 microg/L. The protein, which had a molecular weight of about 70 kD,was purified by Ni-NTA affinity chromatography and identified by SDS-PAGE and Western-blot analysis. Immunofluorescence assay and cellular rosetting showed that it can react specifically on Jurkat (CD3+) and Ramous (CD20+) cells. The lysis of human PBL against CD20-positive lymphoma Ramous cells in the presence of the anti-CD3/anti-CD20 bispecific single-chain antibody can observed by microscope. All these results would lighten the further study of its biological functions in vitro and in vivo.
Animals ; Antibodies, Bispecific ; biosynthesis ; genetics ; immunology ; Antibodies, Monoclonal ; immunology ; Antigens, CD20 ; immunology ; CD3 Complex ; immunology ; CHO Cells ; Cricetinae ; Cricetulus ; Humans ; Immunoglobulin Variable Region ; biosynthesis ; genetics ; immunology ; Lymphoma, B-Cell ; metabolism ; Recombinant Proteins ; Transfection ; Tumor Cells, Cultured

Severe chronic active Epstein-Barr virus (EBV) infection is a rare and life-threatening illness. Although the criteria for diagnosis include chronic or recurrent infectious mononucleosis-like symptoms lasting more than 6 months and high titers of anti-EBV antibodies, clinical and laboratory findings may be heterogeneous and flexible application of those criteria is necessary in cases showing typical clinical and pathologic findings. We report a case of severe chronic active EBV infection in a 62-yr-old female patient who showed classical clinical findings with infiltration of EBV-infected T lymphocytes in the bone marrow, spleen, and lymph nodes, and died four months after presentation.
Antigens, CD3/biosynthesis ; Antigens, CD4/biosynthesis ; Antigens, CD8/biosynthesis ; Bone Marrow Cells/virology ; Epstein-Barr Virus Infections/*diagnosis/*mortality ; Female ; Herpesvirus 4, Human/genetics ; Human ; Immunohistochemistry ; Lymph Nodes/virology ; Lymphocytes/metabolism ; Middle Aged ; Organ Weight ; Spleen/pathology/virology ; Support, Non-U.S. Gov't ; T-Lymphocytes/virology

The term "chordoid meningioma" means meningioma, which is pathologically similar to chordoma, and previously reported that rarely associated with microcytic anemia and/or dysgammaglobulinemia especially in pediatric population. We present a case of this rare variant, which comprises less than 0.5% of all meningiomas. A 33-yr-old man visited our hospital, complaining visual field defect worsening over 7 yr. Neurological examination showed left homonymous hemianopsia. The brain magnetic resonance imaging revealed well enhancing right temporo-occipital mass with cystic portion. Histopathologic findings of resected tumor were compatible with chordoid meningioma which included trabeculae of eosinophilic, vacuolated cells in a myxoid matrix with prominent lymphoplasmacellular infiltration. The neoplastic cells were positive for vimentin and epithelial membrane antigen and negative for glial fibrillary acidic protein and cytokeratin. This is an adult case of chordoid meningioma without anemia or dysgammaglobulinemia.
Adult ; Antigens, CD20/biosynthesis ; Antigens, CD3/biosynthesis ; B-Lymphocytes/pathology ; Brain/pathology ; Brain Neoplasms/*diagnosis/*pathology ; CA-15-3 Antigen/biosynthesis ; Chordoma/*diagnosis/*pathology ; Human ; Magnetic Resonance Imaging ; Male ; Meningeal Neoplasms/*diagnosis ; Meningioma/*diagnosis ; T-Lymphocytes/pathology ; Vimentin/biosynthesis ; Visual Fields

Animals ; Antigens, Neoplasm ; biosynthesis ; genetics ; immunology ; Apoptosis ; physiology ; Breast Neoplasms ; immunology ; metabolism ; CD3 Complex ; immunology ; Female ; Humans ; Killer Cells, Natural ; pathology ; Neoplasms ; immunology ; metabolism ; Receptors, Antigen, T-Cell ; immunology ; Stomach Neoplasms ; immunology ; metabolism ; Uterine Cervical Neoplasms ; immunology ; metabolism

OBJECTIVETo study the influence of percutaneous microwave ablation (PMA) and surgical resection for patients with small primary hepatocellular carcinoma (PHC) on dissemination of tumor cells in peripheral blood determined by AFP mRNA.

METHODSForty patients with small PHC (The maximal diameter < or = 5 cm) confirmed histologically were included in this study. All the patients had single tumor nodule only without metastasis. Of the 40 patients, 19 were treated by PMA and 21 by surgical resection. Blood samples were collected and tested immediately before treatment, 30 min after the mass ablated/resected, 1 d and 7 d later by RTD-Nested-RT-PCR for AFP mRNA. The CD3, CD4, CD8 and CD4/CD8 in blood, and hepatic function were tested at the same time points as well.

RESULTSAfter treatment, ALT and AST in peripheral blood increased in both groups, but more intensely in the surgical group. The CD3, CD4 and CD4/CD8 in peripheral blood decreased at 30 min, 1 day and 7 days after surgical resection, and the lowest value was at 30 min after surgery. The immune function was kept at the same level as pre-treatment in the PMA group. AFP mRNA copies in blood could be detected in 27 of 40 patients (67.5%) in two groups before treatment, and the copy number was increased after treatment. There was no significant difference between the two groups. The patients were followed up for 1 - 16 months. AFP mRNA copies in blood could be detected persistently in the 4 patients with extrahepatic metastasis or liver recurrence.

CONCLUSIONSurgical resection and microwave ablation may cause PHC cells dissemination into the blood circulation in patients with small PHC, and there was no difference between the two treatment groups. The cellular immune function in peripheral blood is decreased after surgical resection, but is maintained at the same level as pre-treatment in the PMA group. The impairment of liver function is less severe after PMA treatment than surgical resection. PMA may provide certain value for clinical management of small hepatocellular carcinoma.

Adult ; Aged ; CD3 Complex ; blood ; CD4 Antigens ; blood ; CD4-CD8 Ratio ; CD8 Antigens ; blood ; Carcinoma, Hepatocellular ; blood ; surgery ; therapy ; Catheter Ablation ; methods ; Female ; Follow-Up Studies ; Hepatectomy ; Humans ; Liver Neoplasms ; blood ; surgery ; therapy ; Male ; Microwaves ; therapeutic use ; Middle Aged ; Neoplasm Recurrence, Local ; RNA, Messenger ; biosynthesis ; genetics ; alpha-Fetoproteins ; biosynthesis ; genetics

OBJECTIVEThe expression of CD25, CD45RA, CD45RO on umbilical cord blood mononuclear cells (CBMCs) and CD3(+) T lymphocytes was investigated to explore the mechanism of immunosuppressive effects of intravenous immunoglobulin on neonatal immune function.

METHODSUmbilical cord blood mononuclear cells and CD3(+) T lymphocytes isolated from 8 neonates were studied. The expression of CD25, CD45RA, CD45RO on umbilical cord blood mononuclear cells (CBMCs) and CD3(+) T lymphocytes induced with various stimuli of different combinations of IVIG and phytohemagglutinin (PHA) including (1) control group, (2) PHA activation group, (3) IVIG pre-inhibition group, (4) PHA pre-activation group, (5) PHA+IVIG group was measured with four-color immunofluorescence antibodies staining-flow cytometric technique. The results were also compared with peripheral blood mononuclear cells of 8 adults (PBMCs).

RESULTSIVIG inhibited the PHA-induced proliferation of CBMCs as reflected by the decreased expression of CD25 and CD45RO. The amounts of CD25(+) and CD4(+)CD45RO(+) CBMCs reached 77.52% +/- 2.31% and 64.29% +/- 3.09% after PHA use. But a decreased response in CD25(+) (7.66% +/- 1.20% and 7.78% +/- 1.46%) and CD4(+)CD45RO(+) CBMC (3.18% +/- 1.90% and 3.11% +/- 0.08%) was observed when IVIG was added in both IVIG pre-inhibition group and PHA+IVIG group. As compared with PBMCs, IVIG failed to induce the increase of the expression of CD45RA in CBMCs whereas CD45RA(+) PBMCs increased from 54.93% +/- 3.63% to 72.77% +/- 0.39% in IVIG pre-inhibition group. Moreover, IVIG inhibited the expression of CD25 and CD45RO on cord blood CD3(+) T lymphocytes no matter whether they were activated with PHA or not. The amounts of CD25(+) and CD4(+)CD45RO(+) CD3(+) T lymphocytes reached 97.92% +/- 2.19% and 80.41% +/- 5.57% after PHA use. But a decreased response in CD25(+) CBMCs (77.29% +/- 0.63%, 51.48% +/- 1.85% and 62.73% +/- 1.24%) and CD4(+)CD45RO(+) CD3(+) T lymphocytes (35.47% +/- 2.55%, 40.14% +/- 1.16% and 36.41% +/- 2.96%) was observed when IVIG was added in IVIG pre-inhibition group, PHA pre-activation group and PHA+IVIG group, and the degree of inhibition of IVIG on cord blood CD3(+) T lymphocytes was much lower than that of CBMCs.

CONCLUSIONSCord blood T lymphocytes activation was inhibited by IVIG through the inhibition of CD25(+) CBMCs expression and the prevention of transformation from CD4(+)CD45RA(+) cells into CD4(+)CD45RO(+) cells. This IVIG-mediated suppression of activation in cord blood T cells may be derived from the indirect effect of other immune cells or molecules other than the direct effects on T cells. IVIG failed to induce the increase of expression of CD45RA in CBMCs, which may be related to the fact that majority of CBMCs were CD45RA(+) cells, but this may not rule out that the immunosuppressive effect of IVIG could be accomplished by the increase of CD45RA(+) cells in adult peripheral blood mononuclear cells. The suppressive effect of IVIG on CD4(+)CD45RO(+) T lymphocytes may account for its inhibitory effect on immunoglobulin production of neonates' B cells. Considering that naïve CD45RA(+) cells dominate in neonates and IVIG can inhibit transformation from CD4(+)CD45RA(+) cells into CD4(+)CD45RO(+) cells, it is recommended that IVIG should be used properly in neonates, otherwise it may deteriorate their poor immune function especially when it is used for prophylaxis or as a treatment of neonatal non-infectious diseases, and its immunosuppressive action will increase the susceptibility of neonates to infection.

Adult ; CD3 Complex ; biosynthesis ; immunology ; Cell Survival ; drug effects ; Cells, Cultured ; Female ; Fetal Blood ; cytology ; immunology ; Flow Cytometry ; Humans ; Immunoglobulins, Intravenous ; administration & dosage ; adverse effects ; immunology ; Immunologic Factors ; adverse effects ; Immunosuppressive Agents ; administration & dosage ; adverse effects ; immunology ; Infant, Newborn ; Injections, Intravenous ; Interleukin-2 Receptor alpha Subunit ; biosynthesis ; immunology ; Leukocyte Common Antigens ; biosynthesis ; immunology ; Leukocytes, Mononuclear ; cytology ; drug effects ; immunology ; Lymphocytes ; cytology ; drug effects ; immunology ; Male