OBJECTIVETo investigate the changes of relative telomere length (RTL) of peripheral blood (PB) CD3⁺, CD3⁺CD4⁺, CD3⁺CD8⁺T lymphocytes, CD19⁺B lymphocytes and bone marrow (BM) CD34⁺ cells and its association with disease severity in untreated patients with immuno-related pancytopenia (IRP).

METHODSThe PB CD3⁺ , CD3⁺ CD4⁺ , CD3⁺ CD8⁺ T lymphocytes, CD19⁺ B lymphocytes, and BM CD34⁺ cells were purified by magnetic activated cell sorting (MACS), and RTL were measured with flow-fluorescence in situ hybridization (FLOW-FISH).

RESULTSThe RTL of CD3⁺, CD3⁺CD4⁺ , and CD3⁺CD8⁺T lymphocytes in untreated IRP patients were (27.754 ± 16.323)%, (7.526 ± 3.745)% and (25.854 ± 14.789)%, respectivly, which were significantly shorter than those in healthy-controls (54.555 ± 19.782)%, (12.096 ± 2.805)%, and (38.367 ± 4.626)% (P<0.05). The RTL of CD19⁺ lymphocytes in untreated IRP patients was (22.136 ± 16.142)%, which was significantly shorter than that in healthy controls (42.846 ± 16.353)% (P<0.01). There was no significant difference of BM CD34⁺ cells RTL between the untreated IRP patients (22.528 ± 21.601)% and the healthy controls (23.936 ± 19.822)% (P>0.05). There were significantly positive correlations between the RTL of B lymphocytes and the count of white blood cell (r=0.706, P=0.015). There were negative correlations between RTL of B lymphocytes and the clinical symptoms (r=-0.613, P=0.045) and positive correlations with therapeutic effect (r=0.775, P=0.005).

CONCLUSIONThe shorter RTL of CD3⁺, CD3⁺CD4⁺, CD3⁺CD8⁺, CD19⁺ lymphocytes, and the normal RTL of BM CD34⁺ cells in untreated IRP patients were identified, which might imply that IRP is a type of acquired autoimmune diseases.

Adolescent ; Adult ; B-Lymphocyte Subsets ; immunology ; Child ; Female ; Humans ; Lymphocytes ; ultrastructure ; Male ; Middle Aged ; Pancytopenia ; immunology ; pathology ; T-Lymphocyte Subsets ; immunology ; Telomere ; ultrastructure ; Young Adult

BACKGROUNDTo study characteristics of peripheral T cell subsets in 94 children with chronic hepatitis B and to elucidate its relationships with clinical status and hepatic pathology.

METHODSPeripheral T cell subsets were detected using flow cytometric analysis with specific monoclonal antibodies staining in 94 patients with HBV infection. The authors simultaneously detected their serum ALT, markers of HBV infection and examined liver biopsy material for pathological changes.

RESULTSIn patients with serious liver lesion, the ratio of CD4+/CD8+ cells was significantly higher than those with mild lesion (1.41+/-0.54 vs 1.08+/-0.35, P less than 0.05), which seemed to be associated with the various liver lesions among the patients. In female cases, the levels of CD4+ T cells and the ratio of CD4+/CD8+ T cells were higher than their counterpart in male cases (33.1+/-5.39 vs 28.8+/-6.28, 1.28+/-0.32 vs 1.02+/-0.36, P less than 0.05), but the level of CD8+ T cells was lower than those in males (26.79+/-4.66 vs 30.51+/-7.17, P less than 0.05). There was no obvious correlation between T cell subsets and circulating HBV viral load, the size of spleen among the HBV-infected children.

CONCLUSIONThe characteristics of peripheral T cell subsets probably suggests the immune disorder occurred in these children with hepatitis B compared with healthy controls and its mechanism needs further investigation.

Child ; Child, Preschool ; Female ; Hepatitis B, Chronic ; immunology ; pathology ; Humans ; Infant ; Liver ; pathology ; Male ; T-Lymphocyte Subsets ; immunology

OBJECTIVETo study of serum level of cortisol and peripheral T lymphocyte subsets state in the hepatitis B virus (HBV) carriers.

METHODSSixty chronic HBV carriers and ten healthy controls were all enrolled in this present study. Serum expression of cortisol was determined by radioimmunoassay, and also flow cytometry was performed to evaluate peripheral blood T lymphocyte subset.

RESULTSCompared with those in normal controls, the serous levels of cortisol in chronic HBV carriers were significantly elevated, while there was no distinct difference in the proportion of CD4+ T lymphocytes ( P > 0.05) with the decreased odds of CD4+/CD8+ lymphocytes( P < 0.05) and obvious higher proportion of CD8+ T lymphocytes( P < 0.05). In comparison between HBeAg positive group and HBeAg negative group, the serous levels of cortisol of the former group were significantly higher ( P < 0.05), and so proportion of CD8+ T was too ( P < 0.05). However, there is no significant differences in the proportion of CD4+ T lymphocyte ( P > 0.05).

CONCLUSIONThe elevated serum cortisol and increased CD8+ T lymphocytes subsets in the chronic HBV carriers, suggested that there was disturbance of endocrine-immune response in the chronicity of HBV infection.

Adult ; Carrier State ; immunology ; pathology ; virology ; Female ; Hepatitis B ; blood ; immunology ; metabolism ; pathology ; Hepatitis B virus ; immunology ; Humans ; Hydrocortisone ; blood ; immunology ; Male ; T-Lymphocyte Subsets ; immunology

OBJECTIVETo measure the quantities and apoptosis-related protein levels of B lymphocyte in the patients with immunorelated pancytopenia (IRP) and explore the action of B lymphocyte in the pathogenic mechanism of IRP.

METHODSQuantities of whole B lymphocytes and CD5+ B lymphocytes as well as the expressions of Fas and Bcl-2 in B lymphocytes in 35 patients with untreated IRP, 15 IRP patients in complete remission (CR), and 10 normal controls were assayed by flow cytometry.

RESULTSThe percentages of B lymphocyte and CD5+ B lymphocyte were significantly higher in untreated IRP patients than in CR IRP patients and normal controls (P < 0.05), and there was no significant difference between the latter two groups (P > 0.05). There was no significant difference of Fas expression in B lymphocyte among three groups (P > 0.05). The expression of Bcl-2 in B lymphocyte was significantly higher in untreated patients than in CR patients or normal controls (P < 0.01), and significantly higher in CR patients than in normal controls (P < 0.01). The apoptosis-related index was significantly lower in untreated patients than in CR patients or normal controls (P < 0.05), and significantly lower in CR patients than in normal controls (P < 0.05). The percentage of B lymphocyte was positively correlated with post-treated response time (r = 0.53, P < 0.01).

CONCLUSIONThe production of auto-antibodies in IRP patients probably has some relationship with the abnormal quantities of B lymphocyte and its subpopulations as well as with the inhibition of B lymphocyte apoptosis.

Adolescent ; Adult ; B-Lymphocytes ; cytology ; immunology ; Case-Control Studies ; Child ; Female ; Humans ; Lymphocyte Subsets ; Male ; Middle Aged ; Pancytopenia ; immunology ; pathology

An orthotopic penetrating keratoplasty model was developed in the rat. An oversized (0.5 mm) graft was used and 8 interrupted sutures were applied. These sutures were not removed. Eleven grafts out of 13 were rejected by the 3rd week in the disparate group (Brown Norway rat to Lewis rat transplantation group), which was characterized by edema, opacity, and neovascularization. All grafts remained clear in the syngeneic group (Lewis rat to Lewis rat transplantation group). Immunohistochemical examination was performed. This model seems to be a reliable and reproducible one to evaluate rejection reaction in corneal transplantation.
Animals ; Female ; Graft Rejection ; Keratoplasty, Penetrating/immunology/*methods/pathology ; Lymphocyte Subsets/immunology/pathology ; Macrophages/immunology/pathology ; Rats ; Rats, Inbred BN ; Rats, Inbred Lew ; Transplantation, Homologous ; Transplantation, Isogeneic

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

OBJECTIVETo investigate the dynamic changes of lymphocyte subsets before and after autologous hemopoietic stem cell transplantation (HSCT) in severe/refractory autoimmune disease (AID) and study the post-transplantation immunological reconstitution in AID.

METHODSThirteen patients with severe/refractory AID who registered for HSCT from April 2003 to April 2005 in Peking Union Medical College Hospital, including 8 patients with systemic lupus erythematosus, 4 patients with rheumatoid arthritis, and 1 patient with primary Sjögren's syndrome (pSS) were enrolled in this study. Blood samples were collected before/after mobilization, before conditioning, and 2 weeks, 1 month, 3 months, 6 months, 12 months, and 18 months post-transplantation. Lymphocyte subsets were tested by flow cytometry as follows: T cell (CD3 +), B cell (CD19 +), natural killer (CD3-CD16 + CD56 +), Th (CD3 + CD4 +), Tc (CD3 + CD8 +), naïve T (CD4 + CD45RA), memory T (CD4 + CD45RO), and CD4/CD8 ratio.

RESULTSLymphocyte subsets for SLE patients were severely abnormal compared to normal or RA patients (both P < 0.01). B cell reconstituted to normal level within 18 months, meanwhile NK and T cell remained low. The repopulations of Th and naive T cell were delayed, which caused the up-side-down of CD4/CD8 ratio and low level of naYve T cell percentage for a relatively long time.

CONCLUSIONSLymphocyte subsets abnormality in SLE patients are more severe than in RA patients. Although most autoimmune T/B cell in the grafts and patients can be effectively removed after transplantation, nonmyeloablative conditioning may be a risk for the relapse of AID. The long-term inhibition of CD4 + T cell may be related with the relief of AID after transplantation.

Arthritis, Rheumatoid ; blood ; immunology ; therapy ; Hematopoietic Stem Cell Transplantation ; Humans ; Lupus Erythematosus, Systemic ; blood ; immunology ; therapy ; Lymphocyte Subsets ; immunology ; pathology ; Sjogren's Syndrome ; immunology ; therapy

BACKGROUNDTo explore the pathological features and pathogenesis of severe acute respiratory syndrome (SARS) to provide evidence for the clinical treatment and prevention of SARS.

METHODSPathological features of 2 cases of full autopsy and 4 cases of needle biopsy tissue samples from the patients who died from SARS were studied by light and electron microscopy. The distribution and quantity of lymphocyte subpopulations in the lungs and immune organs from SARS patients were analyzed by immunohistochemistry. The location and semi-quantitative analysis of SARS coronavirus in the tissue specimens were studied by electron microscopy, in situ hybridization and immunohistochemistry.

RESULTSIn total of 6 cases, diffuse alveolar damage and alveolar cell proliferation were common. The major pathological changes of 2 autopsy cases of SARS in lung tissues were acute pulmonary interstitial and alveolar exudative inflammation, and 2 autopsy and one biopsy lung tissues showed alveolar hyaline membrane formation. Terminal bronchiolar and alveolar desquamation of lung tissues in one autopsy and 2 biopsy cases were noted. Among 6 cases, 2 biopsy cases presented early pulmonary fibrosis and alveolar organization. Meanwhile, the immune organs, including lymph nodes and spleens from 2 autopsy cases of SARS whose disease courses were less than 12 days showed extensive hemorrhagic necrosis, reactive macrophage/histocyte proliferation, with relative depression of mononuclear and granulocytic clones in the bone marrows. However, spleen and bone marrow biopsy tissue samples from 4 dead SARS cases whose clinical course lasted from 21 to 40 days presented repairing changes. SARS coronaviruses were mainly identified in type I and II alveolar epithelia, macrophages, and endothelia; meanwhile, some renal tubular epithelial cells, cardiomyocytes, mucosal and crypt epithelial cells of gastrointestinal tracts, parenchymal cells in adrenal glands, lymphocytes, testicular epithelial cells and Leydig's cells were also detected by electron microscopy combined with in situ hybridization. The semi-quantitative analysis of lymphocyte subpopulations revealed that the proportion of CD8+ T lymphocytes were about 80% of the total infiltrative inflammatory cells in the pulmonary interstitium, with a few CD4+ lymphocytes CD3+, CD4+, CD8+ or CD20+ lymphocyte subpopulations were obviously decreased and there was imbalance in number and proportion, while CD57+, CD68+, S-100+ and HLA-DR+ cells were relatively increased in lymph nodes and spleens.

CONCLUSIONSHistologically, the pulmonary changes could be divided into acute inflammatory exudative, terminal bronchiolar and alveolar desquamative and proliferative repair stages or types during the pathological process of SARS. SARS coronavirus was found in multi-target cells in vivo, which means that SARS coronavirus might cause multi-organ damages which were predominant in lungs. There were varying degrees of decrease and imbalance in number and proportion of lymphocyte subpopulations in the immune organs of the patients with SARS. However, these changes may be reversible. It was found that cellular immune responses were predominant in the lungs of SARS cases, which might play an important role in getting rid of coronaviruses in infected cells and inducing immune mediated injury.

Aged ; Female ; Humans ; Lung ; immunology ; pathology ; virology ; Lymphocyte Subsets ; immunology ; Male ; Middle Aged ; SARS Virus ; isolation & purification ; ultrastructure ; Severe Acute Respiratory Syndrome ; immunology ; pathology ; virology

Immune mediated suppression of hematopoiesis has been considered as one of the most important mechanisms leading to pancytopenia in myelodysplastic syndromes. This research was aimed at evaluating immune state of the MDS patients, analyzing the peripheral blood T cell subsets and CD3zeta chain expression and searching the possible reasons of hematopoietic disorders in 11 cases of MDS. Peripheral blood mononuclear cells were collected from 11 patients whose diagnosis was confirmed according to the new WHO diagnostic criteria. Flow cytometry was used for the counts of IFNgamma(+)CD4(+) cell (Th1), IL4(+)CD4(+) cell (Th2), IFNgamma(+)CD8(+) cell (Tc1), and IL4(+)CD8(+) cell (Tc2), and for the analysis of expression of CD3zeta chain in T cell subsets. The results showed that CD8(+) cells increased significantly in MDS patients; there was no significant difference between Th1/Th2, Tc1/Tc2 ratios of T cell subsets and normal control; CD3zeta chain, the functional protein in the signal transduction pathway of T cell, was over expressed in the CD8(+) cell. In conclusion, research indicates that abnormal changes of T cell subgroups exist in peripheral blood of MDS patients. Enhancement of CD8(+) cells and over-expression of CD3zeta chain are important features, which suggest that CD8(+) cells play the most critical role in the pathologic process as compared with other T cell subsets. The over active immunity mediated by T cell subset may be one of the major mechanisms resulting in cytopenia in MDS.
Aged ; Aged, 80 and over ; CD3 Complex ; biosynthesis ; CD8-Positive T-Lymphocytes ; immunology ; metabolism ; pathology ; Female ; Flow Cytometry ; Humans ; Lymphocyte Count ; Male ; Myelodysplastic Syndromes ; immunology ; metabolism ; pathology ; T-Lymphocyte Subsets ; immunology ; metabolism ; pathology

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