It was long believed that platelets are prematurely destroyed in the reticuloendothelial system by platelet autoantibodies in idiopathic thrombocytopenic purpura. However, humoral mechanisms cannot account for all observations made in this disorder, and it is increasingly evident that cellular mechanisms contribute to platelet destruction. In this review the tolerance of T cell, abnormality of T cell apoptosis, abnormal activation of T cells, T cell subtype and its function changes, and T cell-mediated cytotoxicity were summarized.
Apoptosis ; immunology ; Autoantibodies ; immunology ; Humans ; Immune Tolerance ; Immunity, Cellular ; immunology ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; T-Lymphocyte Subsets ; immunology ; T-Lymphocytes ; immunology ; pathology ; T-Lymphocytes, Cytotoxic ; immunology ; T-Lymphocytes, Helper-Inducer ; immunology

OBJECTIVETo induce anti-B-cell acute lymphoblastic leukemia (B-ALL) cytotoxic T lymphocyte response against immunoglobulin heavy chain frame-derived nonapeptide.

METHODSThe peptide, QLVQSGAEV, containing IgHV1 frame region 3rd-11th amino acids (IgHV1(3-11)), was synthesized. IgHV1(3-11)-T2 binding tests were performed. HLA-A * 0201-positive normal peripheral blood mononuclear cells (PBMNC) were stimulated by IgHV1(3-11)-loaded antigen presenting cells three times at weekly intervals. HLA-A * 0201/IgHV1(3-11) tetramers were used to detect the proliferation of IgHV1(3-11)-specific CD8(+) T cells in the culture. Seven IgHV gene families of B-ALL patients were respectively amplified by PCR and the PCR products were sequenced to select IgHV1 and IgHV3 family monoallelic functional rearrangements. Among them, HLA-A * 0201 positive individuals were subsequently identified. Cytotoxicity of IgHV1(3-11)-specific CD8(+) T cells against HLA-A * 0201-positive IgHV1/IgHV3 family B-ALL cells was measured by MTT assay.

RESULTSThe synthesized IgHV1(3-11) up-regulated HLA-A * 0201 expression on T2 cell surface by 1.63-folds. The percentage of IgHV1(3-11)-specific CD8(+) T cells in HLA-A * 0201-positive normal PBMNC was increased from 1.64% after second stimulation to 82.57% after third stimulation. At effector: target ratio of 20:1, the killing rate of IgHV1(3-11)-specific CD8(+) T cells against IgHV1 family B-ALL cells was 18.24%, being 1.8-folds as that against IgHV3 family B-ALL cells (P = 0.01).

CONCLUSIONCytolytic T lymphocytes generated in vitro against immunoglobulin heavy chain frame-derived nonapeptides can kill B-ALL cells expressing these peptides.

Cells, Cultured ; Humans ; Immunoglobulin Heavy Chains ; immunology ; pharmacology ; Leukemia, B-Cell ; immunology ; pathology ; T-Lymphocytes, Cytotoxic ; drug effects ; immunology

This study was aimed to investigate the effects of tumor antigen-loaded dendritic cells (DC) stimulating the specific cytotoxic T lymphocytes (CTL) on Jurkat cells in vitro. Peripheral blood mononuclear cells were isolated by Ficoll density gradient centrifugation from normal human heparinized blood, the adherent monocytes were cultured with granulocyte-macrophage colony stimulating factor (GM-CSF), interleukin-4 (IL-4), alpha tumor necrosis factor (TNF-alpha) and sCD40L, DCs were co-cultured with frozen-thawed antigen of Jurkat cells or WT1 peptides, and then T cells were triggered into specific CTL. The results showed that most suspended cells exhibited distinctive morphological features of DC which expressed CD40 (96%), CD86 (97%), CD80 (77%), CD1a (69%), and gained the powerful capacity to stimulate proliferation of allogeneic lymphocytes. Under the effector: target ratio of 20:1, CTLs derived from cultures with DC and frozen-thawed antigen of Jurkat cells showed 91.1% cytotoxicity against Jurkat cells, CTL derived from cultures with DC and WT1 peptides showed 87.5% cytotoxicity against Jurkat cells, cytotoxicity of CTL derived from cultures with unloaded DC against Jurkat cells was 42.1% and cytotoxicity of monocytes was 22.7%. Cytotoxicity of CTL derived from culture with frozen-thawed antigen or WT1 peptides loaded DC was stronger than that in control groups (P < 0.01). It is concluded that the tumor antigen-pulsed DC can induce efficient and specific anti-tumor immunity, may play a great role in clinical therapy for leukemia.
Antigens, Neoplasm ; immunology ; Cells, Cultured ; Coculture Techniques ; Dendritic Cells ; cytology ; immunology ; Humans ; Jurkat Cells ; Leukemia, T-Cell ; immunology ; pathology ; Lymphocyte Activation ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Cells, Cultured

The pathogenesis of some autoimmune diseases has been considered to be related to abnormal differentiation of T cell subsets. This study was aimed at investigating the change of Th1-like and Th2-like cells balance in ITP children, and analyzing the role of T cell subsets disequilibrium in the pathogenesis of ITP. Peripheral blood T cells were collected from 30 ITP patients, the T-cells were isolated and purified. The ratios of Th/Tc, Th1/Th2 and Tc1/Tc2 in peripheral blood T cells were analyzed by immunofluorescence staining and bicolor flow cytometry (FCM) in vitro. The results showed that as compared with the ratios of Th1/Th2 (48.76% +/- 6.17%) and Tc1/Tc2 (18.90% +/- 4.12%) in healthy children, the ratios of Th1/Th2 (56.21% +/- 5.95%) and Tc1/Tc2 (23.09% +/- 3.31%) in ITP children increased obviously. FCM analysis revealed that average percentages of Th, Th1, Th2, Tc, Tc1 and Tc2 were 22.31% +/- 6.51%, 21.92% +/- 6.42%, 0.39% +/- 0.14%, 31.12% +/- 6.15%, 30.95% +/- 5.45% and 1.34% +/- 0.84% in ITP children versus 39.24% +/- 5.82%, 39.01% +/- 5.47%, 0.80% +/- 0.16%, 30.25% +/- 5.63%, 28.72% +/- 5.20% and 1.52% +/- 0.68% in healthy children. The average percentages of Th, Th1 and Th2 decreased obviously, while the average percentages of Tc, Tc1 and Tc2 did not change. It is concluded that the ratios of Th1/Th2 and Tc1/Tc2 in peripheral blood T cells increase obviously in ITP children and the cellular immunity in ITP children shifts to Th1 type immunity superiority, which suggest that the abnormal differentiation of T cell subsets may play an important role in the pathologic process of ITP.
Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; T-Lymphocyte Subsets ; immunology ; metabolism ; pathology ; T-Lymphocytes, Cytotoxic ; chemistry ; Th1 Cells ; immunology ; Th2 Cells ; immunology

Animals ; Cancer Vaccines ; immunology ; Cell Fusion ; methods ; Cell Line ; Dendritic Cells ; cytology ; immunology ; Fibrosarcoma ; immunology ; pathology ; Hybrid Cells ; immunology ; Mastocytoma ; immunology ; pathology ; Mice ; Mice, Inbred BALB C ; T-Lymphocytes ; immunology ; pathology ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Cells, Cultured

After having successfully constructed and expressed the gene of the anti-CD3/anti-CD20 bispecific single-chain antibody (bscCD3 x CD20), here we analyzed its in vitro bioactivity of mediating the lysis of Ramous human B-lymphoma cells in the presence of T-enriched human peripheral blood lymphocytes (PBL). Obvious opoptosis characters were observed by Annexin V/PI(AV/PI) stained and scanning electron microscope. As evaluated by non-radioactive cytotoxity assay, the bscCD3 x CD20 showed potent bioactivity of mediating human B-lymphoma cells lysis in the presence of T-enriched human PBL. The potency of cytotoxicity depended on the ratios of effect cells to target cells (E:T) used. Further, the antibody showed a dose and time-dependent effect on mediating Ramous cells lysis. The specific lysis reached about 87.3% at an antibody concentration of 5microg/mL and E:T used at 10:1. Clear changes in apoptogenes expression profiles were detected by apoptosis gene array after Ramous cells were treated with the antibody and PBL. Among the upregulated apoptogenes, ATM and P53 showed an increase of 187 times and 15 times respectively, which suggested that ATM-p53 pathway may be the main apoptosis way of Ramous cells induced by T cells in the presence of the bscCD3 x CD20.
Antibodies, Bispecific ; immunology ; Antigens, CD20 ; immunology ; Apoptosis ; immunology ; CD3 Complex ; immunology ; Humans ; Lymphoma, B-Cell ; immunology ; pathology ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Cells, Cultured

OBJECTIVEHuman Ewing sarcoma A673 cells and human peripheral blood-derived DCs were fused to induce an antitumor activity against human EW.

METHODSEW A673 cells and human peripheral blood-derived DCs were fused with polyethylene glycol (PEG).

RESULTSMature DCs with highly expressed surface markers (CD80, CD86, CD83 and HLA-DR) were generated in vitro and flow cytometry. It showed that the highest fusion efficiency was 23.01%. T cell proliferation assay indicated that the novel dendritomas in fused DCs/A673 cells were the most potent in activation of autologous T cell proliferation. The IFN-gamma assay showed that The IFN-gamma secretion by CTLs activated by the novel dendritomas increased more than by other stimulators. CTL assay demonstrated that the novel dendritomas induced A673 cell-specific cytotoxic responses to lyse the A673 cells in the context of MHC class I.

CONCLUSIONThe data indicates that fusion of tumor cells with DCs is an attractive strategy to induce tumor rejection.

Cancer Vaccines ; immunology ; Cell Fusion ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; Humans ; In Vitro Techniques ; Sarcoma, Ewing ; immunology ; pathology ; T-Lymphocytes, Cytotoxic ; immunology

Dendritic Cells ; immunology ; Granzymes ; Humans ; Immune Tolerance ; Killer Cells, Natural ; immunology ; Multivariate Analysis ; S100 Proteins ; analysis ; Serine Endopeptidases ; analysis ; Stomach Neoplasms ; immunology ; pathology ; T-Lymphocytes, Cytotoxic ; immunology

To investigate the biological properties of cryopreserved dendritic cell (DC) derived from K562 cell line, thus to provide a simple, quick and efficient preservative method of DC for infusion of DC to patients with leukemia after complete remission, fresh DC induced from K562 cell line (K562-DC) was frozen in -196 degrees C liquid nitrogen and -80 degrees C mechanical freezer by method of steps (RPMI 1640 with 10% DMSO, 20% FCS as cryopreservatives), and thawed in different time, respectively. Survivals of cryopreserved and fresh K562-DC, expression of surface antigens, stimulating index (SI) and cytotoxic eliminating rate were detected. The results of fresh induced cells were compared with that of cryopreserved ones. The results showed that before and after frozen in liquid nitrogen, the morphological characteristics of K562-DC had no distinct change; and both their expression rates of surface molecular and capacity to stimulate allogeneic lymphocyte had no statistic significance (P > 0.05). In addition, there were no differences in terms of viability, stimulatory capacity and cytotoxicity of K562-DC from two ways for less than one month (P > 0.05), but there were differences when frozen for more than one month (P < 0.05). It is concluded that there is no significant difference when frozen less than one month between liquid nitrogen and -80 degrees C freezer; but when time is more than one month, K562-DC frozen in -196 degrees C liquid nitrogen is better than that in -80 degrees C freezer.
Antigens, Surface ; immunology ; Cell Shape ; Cell Survival ; Cells, Cultured ; Cryopreservation ; methods ; Dendritic Cells ; immunology ; pathology ; Humans ; K562 Cells ; T-Lymphocytes, Cytotoxic ; immunology ; pathology ; Time Factors

Most of thyroid lymphomas are B-lineage, and T-cell lymphomas are rare. Here, we report a case of primary thyroid T-cell lymphoma associated with Hashimoto's thyroiditis. A 48-yr-old woman presented with incidentally found neck mass. Histologically, the resected right lobe of the thyroid was replaced by monomorphic small atypical lymphoid cells with lymphoepithelial lesion-like change, most of which were immunoreactive for CD3, CD8, betaF-1, and TIA-1. Peripheral T-cell lymphoma, unspecified, was finally diagnosed after molecular study for TCR-gamma gene rearrangement. This is the second case of cytotoxic T-cell lymphoma reported in the thyroid gland so far. Unique association between thyroid follicles and neoplastic lymphocytes may be characteristic feature of this type of T-cell lymphoma.
Female ; Hashimoto Disease/*pathology ; Humans ; Lymphoma, B-Cell, Marginal Zone/*pathology ; Lymphoma, T-Cell/*pathology ; Middle Aged ; T-Lymphocyte Subsets/immunology/pathology ; T-Lymphocytes, Cytotoxic/immunology/pathology ; Thyroid Gland/*pathology