OBJECTIVETo detect memory B lymphocyte (Bm) in peripheral blood (PB) of immune-related pancytopenia (IRP).

METHODS86 patients with IRP and 11 health volunteers were enrolled in this study. Bm (CD5⁺ CD19⁺ CD27⁺) and bone marrow mononucleated cell antibodies (BMMNC-Ab) were determined via fluorescence-activated cell sorting, and clinical outcomes of these patients were analyzed.

RESULTS(1)43 initial patients achieved obvious remission in all 52 initial cases after conventional immunosuppression therapy. 16 relapsed patients with IRP received Rituximab (RTX) and 14 cases achieved obvious remission, among which 7 cases were refractory to conventional immunosuppression therapy, 5 cases exhibited obvious remission, and 2 cases did not respond. Other 18 relapsed cases received conventional immunosuppression therapy and 13 cases achieved obvious remission. (1)The level of Bm in PB in 52 initial patients with IRP was(1.81 ± 0.97)%, and no significant difference was observed between the initial patients and health volunteers (1.75 ± 0.55)% (P>0.05). The level of Bm in PB in 34 relapsed patients with IRP was obviously higher than that in the initial IRP patients and health volunteers (P<0.05). Significant difference was observed in the level of Bm in PB in 16 relapsed IRP patients between pre-therapy and post-therapy with RTX (P<0.05). No statistical difference was found between the remission and no-response groups in relapsed patients treated with RTX. RTX regimen produced more effective outcome than conventional immunosuppression therapy, which better eliminated Bm than the latter (P<0.05). Initial patients with IRP who relapsed within a two-year follow-up period had a lower level of Bm in PB compared with un-relapsed patients (P<0.05). Majority of BMMNC- Ab antibodies in relapsed patients were IgG (82.4%) and IgM (69.2%) autoantibodies in patients with initial IRP.

CONCLUSIONThe level of Bm in PB was associated with relapsed patients with IRP. Bm did not respond to conventional immunosuppression therapy,but responded to RTX.

Adolescent ; Adult ; Antibodies, Monoclonal, Murine-Derived ; therapeutic use ; B-Lymphocyte Subsets ; immunology ; Female ; Humans ; Immunologic Memory ; Immunosuppression ; Male ; Middle Aged ; Pancytopenia ; immunology ; therapy ; Recurrence ; Rituximab ; Treatment Outcome ; Young Adult

BACKGROUNDRegulatory T cells (Tregs) may play an important role in immunopathology during HIV-1 infection. Transcription factor forkhead box P3 (FoxP3) orchestrates the development of Tregs and is a useful marker to identify this population. Using a FoxP3 phenotype to define Tregs, we investigated the level and phenotype of peripheral blood natural CD4(+)Tregs and assessed the relationship between the frequencies and absolute numbers of CD4(+) Tregs and disease progression among untreated HIV-infected men who have sex with men (HIV(+) MSM) in China.

METHODSFifty-two untreated HIV(+) MSM with CD4(+) T-cell counts of ≤ 350 cells/µl or > 350 cells/µl were compared in a cross-sectional study. Twelve age-matched HIV-uninfected MSM and nine patients receiving antiretroviral therapy for at least 1 year were also included. Expression of CD25, CD127, CD45RA, CCR7 and CTLA-4 was assessed on CD4(+) Tregs using polychromatic flow cytometry.

RESULTSThe percentage of CD4(+) Tregs was increased significantly, whereas CD4(+) Tregs expressed less CTLA-4 in HIV(+) MSM compared with controls. CD4(+) Tregs displayed predominantly an effector memory phenotype (CD45RA(-) CCR7(-)), phenotypically distinct from conventional CD4(+) T cells. Moreover, the expansive frequencies of CD4(+) Tregs coincided with lower CD4(+) T-cell counts and higher viral loads whereas the absolute numbers of CD4(+) Tregs were associated with higher CD4(+) T-cell counts and lower viral loads. The expansion of Tregs was also associated with CD8(+) T-cell activation.

CONCLUSIONIncreased proportions and decreased numbers of CD4(+) Tregs are associated with HIV progression, and their functions may impair with the progression of HIV infection.

Acquired Immunodeficiency Syndrome ; immunology ; Adult ; CD4 Lymphocyte Count ; CTLA-4 Antigen ; analysis ; Cross-Sectional Studies ; Disease Progression ; HIV-1 ; Homosexuality, Male ; Humans ; Immunologic Memory ; Lymphocyte Activation ; Male ; Middle Aged ; RNA, Viral ; blood ; T-Lymphocytes, Regulatory ; immunology

OBJECTIVESTo study the effect of HAART on subsets of T lymphocytes and expression of CD127 on memory and naїve CD4(+) and CD8(+)T cells in pediatric AIDS patients with different viral loads receiving HAART.

METHODA cross- sectional study on 194 pediatric AIDS patients receiving HAART was carried out and 52 age matched healthy children were recruited as controls. The percentage of CD4(+), CD8(+), CD8(+)CD45RA(+)CD127(+/-), CD8(+)CD45RO(+)CD127(+/-), CD4(+)CD45RA(+)CD127(+/-) and CD4(+)CD45RO(+)CD127(+/-)T cells was tested using flow cytometry, and HIV-RNA in plasma was detected by quantitative RT-PCR.

RESULTThe percentage of memory (CD45RO(+)) CD4(+)T cells decreased to (45.73 ± 8.85)%, and that of naїve (CD45RA(+)) CD4(+) and memory CD8(+)T increased to (60.44 ± 5.01)% and (54.69 ± 7.71) % respectively in the pediatric AIDS patients vs. controls (P < 0.05). The percentage of naїve (CD45RA(+)) CD4(+)T cells of patients with viral load (VL) < 400 copies/ml was (65.57 ± 5.33) %, which was significantly higher than that of patients with VL ≥ 400 copies/ml (P < 0.05).Of patients with VL < 400 copies/ml, the percentage of CD4(+)CD127(+)T cells, especially the subset of memory CD4(+)CD127(+)T cells was (82.35 ± 2.31)%, which was higher than that of patients with VL ≥ 400 copies/ml, but lower than that of controls (P < 0.05). The percentage of memory and naїve CD8(+)CD127(+)T cells was lower than that of controls (P < 0.05).

CONCLUSIONThe recovery of CD4(+)T cell subsets in pediatric AIDS patients is associated with viral load. Effective HAART can increase the percentage of naїve CD4(+)T cells and the life of memory CD4(+)T cells.

Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; virology ; Adolescent ; Antiretroviral Therapy, Highly Active ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Child ; Cross-Sectional Studies ; Female ; Flow Cytometry ; Humans ; Immunologic Memory ; Interleukin-7 Receptor alpha Subunit ; immunology ; metabolism ; Lymphocyte Count ; Male ; Reverse Transcriptase Polymerase Chain Reaction ; T-Lymphocyte Subsets ; immunology ; Viral Load

OBJECTIVEThe aim of the study was to evaluate the anti-HBs persistence and the long term preventive efficacy after vaccination 23 years with plasma-derived hepatitis B vaccine.

METHODSThe study consisted of 261 children who were 5 - 9 years aged, from two primary schools in two townships of Xi'an. 126 children were randomly selected as vaccine group, and 135 children in control group. These children were followed up again in 2009. Excluding self-inoculation, the vaccine and control groups were 81 and 75, who was used to ask to recall details of their experience for vaccination and liver-related illnesses during past twelve years. Individuals who had anti-HBs titers less 10 mIU/ml, HBsAg, anti-HBc and HBV-DNA all were negative, were given a booster dose vaccine and retest for anti-HBs titer after one month.

RESULTSAfter eliminated the interference of an early booster dose and vaccination outside the study, the positive rate of anti-HBs was 48.1% (39/81) in the vaccine group at year 23, higher than 34.7% (26/75) in control group. At year 23 after primary vaccination, 84.0% (21/25) individuals in the vaccine group whose anti-HBs and anti-HBc both are negative showed a stronger anamnestic response after received a booster dose, while 7.5% (3/40) in the control group. At year 23 after primary vaccination, none clinical case of hepatitis B was found among 194 individuals. However, anti-HBc positive rate in the vaccine group was 16.0% (13/81), while the rate in the control group was 30.7% (23/75) (χ(2) = 4.687, P < 0.05).

CONCLUSIONAt 23 years after implemented a full course of plasma-derived hepatitis B vaccine, the recipients of vaccine were maintained anti-HBs at a high level or strong immunological memory.

Child ; Child, Preschool ; Follow-Up Studies ; Hepatitis B ; immunology ; prevention & control ; Hepatitis B Antibodies ; blood ; Hepatitis B Vaccines ; immunology ; Humans ; Immunization, Secondary ; Immunologic Memory ; immunology ; Plasma ; immunology

OBJECTIVETo investigate the immunization status of hepatitis B vaccine who were inoculated at birth, HBV infections and the vaccine booster effect in the first-year middle school students (12 - 14 years old).

METHODSA cluster, stratified simplified random sampling method was administrated. The sample size was at least 218, which was calculated by Epi Info 3.3.2 software at 53% the minimum acceptable anti-HBs positive rate and 95% confidence level. A total of 250 and 236 students participated in the infection status and booster immunization effects investigation. The HBsAg, anti-HBs and anti-HBc IgG were detected by Enzyme-linked immunosorbent assay (ELISA). HBV DNA was detected by fluorescence quantitative PCR, and the diagnostic test kit were produced respectively by ABBOTT, Diasorin and Beijing Wantai Biological Pharmacy Enterprise Co.

RESULTSFor the immunization status before booster: the positive rate of anti-HBs was 62.80% (157/250), the GMT was 73.79 IU/L; the currently HBV infection rate (HBsAg and anti-HBc positive) was 2.80% (7/250). After injection, the anti-HBs positive rate was 94.92% (224/236). Compared with the before booster results, the significant difference was observed (χ(2) = 73.92, P = 0.00). The GMT was 521.15 IU/L, comparing with the before booster results, there was significant difference (t = 15.98, P = 0.00). The anti-HBs conversion rate (from negative to positive) was 91.86% (79/86) after immune-enhancement; of which, 11 students got the second dose of booster vaccine who are no-responders after first injection, in addition 8 students got the anti-HBs.

CONCLUSIONIt is an effective method to put the first-year middle school students into the immune-enhancement program, so as to improve the immunization memory effect and avoid the loss of protective antibodies.

Adolescent ; Child ; China ; Dose-Response Relationship, Immunologic ; Female ; Hepatitis B ; immunology ; prevention & control ; Hepatitis B Antibodies ; immunology ; Hepatitis B Vaccines ; administration & dosage ; immunology ; Humans ; Immunization, Secondary ; Immunologic Memory ; immunology ; Male ; Schools ; Students

BACKGROUNDThe role of Wilms' tumor 1 protein (WT1)-specific cytotoxic T cells (CTL) in eradicating chronic myeloid leukemia (CML) cells is to be established. The aim of this study was to determine whether WT1 contributed to the graft-versus-leukemia effects (GVLE) for CML following allogeneic hematopoietic stem cell transplantation (HSCT).

METHODSHigh-resolution human leukocyte antigen (HLA) class I genotyping was performed by sequence-specific polymerase chain reaction (PCR). Fifteen HLA-A*2402 patients with CML who underwent allogeneic HSCT were enrolled in this study. We monitored the frequency of WT1-specific CTL by pentamer assay and the molecular minimal residual disease by real-time quantitative PCR.

RESULTSA CD8(+) T-cell response to WT1 was observed in 14 of 15 patients after HSCT. The median frequencies of WT1-CTL were 0.54%, 0.62%, 0.81% and 1.28% (%CD8) on days 30, 60, 90 and 180, respectively. The median frequency of WT1-CTL (1.38%) in patients with molecular remission (MoR) was significantly higher than that in those without MoR (0.38%) on day 30, while no significant differences between them were detected on days 60, 90 and 180. The increase of WT1-CTL was associated with a decrease in bcr-abl expression and MoR; and the decrease of WT1-CTL was associated with an increase in bcr-abl expression, suggesting a WT1-driven GVL effect. WT1-CTL had a predominant effector-memory phenotype (CD45RO(+)CD27(-)CD57(+)).

CONCLUSIONSThe emergence of WT1-CTL with an effector-memory phenotype is associated with GVLE in CML patients after HSCT. This will pave the way for the WT1 vaccines to enhance GVLE after HSCT in CML.

Adult ; CD8-Positive T-Lymphocytes ; immunology ; Female ; Genotype ; HLA Antigens ; genetics ; Hematopoietic Stem Cell Transplantation ; Humans ; Immunologic Memory ; Immunophenotyping ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; immunology ; therapy ; Male ; Middle Aged ; Polymerase Chain Reaction ; T-Lymphocytes, Cytotoxic ; immunology ; metabolism ; WT1 Proteins ; metabolism ; Young Adult

Adult ; Alanine Transaminase ; blood ; CD4-Positive T-Lymphocytes ; immunology ; metabolism ; CD8-Positive T-Lymphocytes ; immunology ; metabolism ; Carrier State ; immunology ; virology ; DNA, Viral ; blood ; Female ; Flow Cytometry ; Hepatitis B ; immunology ; virology ; Hepatitis B virus ; immunology ; Humans ; Immunologic Memory ; Interleukin-7 Receptor alpha Subunit ; immunology ; metabolism ; Male

OBJECTIVETo investigate effect of tumor-specific T cell receptor gene transfection on memory T cell differentiation in vitro.

METHODSTCRVbeta7.1 gene was transferred into peripheral blood mononuclear cells (PBMCs) obtained from healthy adults, and the expression of Vbeta7.1 was detected by flow cytometry before and after the transfection. Memory T cell differentiation was induced by stimulation with the hepatocarcinoma cell line BEL-7402 in vitro. The expression of surface molecules CD45RO, CD45RA and CCR7 was analyzed by flow cytometry to identify the phenotype and subsets of the memory T cells. Fluorescence-activated cell sorting was performed to detect the apoptosis of the tumor cells, and enzyme-linked immunoabsorbent assay was used to determine the production of interferon-gamma (IFN-gamma) for assessing the immune function of the memory T cells.

RESULTSFlow cytometry showed that TCRVbeta7.1 gene was efficiently expressed after transfection. After stimulation by the tumor cells in vitro, the expression of CD45RO in TCRVbeta7.1 gene-modified T cells increased gradually, and analysis of the coexpression of CD45RA and CCR7 revealed that the effector memory T cells constituted the majority of the differentiated memory T cells. The apoptotic rate of the tumor cells induced by the T cells increased significantly with also obviously increased INF-gamma secretion in the memory T cells.

CONCLUSIONTumor-specific TCRVbeta7.1 gene transfection can promote the differentiation of the memory T cells, the majority of which belongs to effector memory T cells that perform immune functions by inducing apoptosis and cytokine secretion.

Adult ; Apoptosis ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Differentiation ; Cell Line, Tumor ; Cells, Cultured ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Genes, T-Cell Receptor alpha ; genetics ; Humans ; Immunologic Memory ; immunology ; Interferon-gamma ; metabolism ; Leukocyte Common Antigens ; metabolism ; Liver Neoplasms ; genetics ; metabolism ; pathology ; T-Lymphocytes ; cytology ; immunology ; metabolism ; Transfection

OBJECTIVESProgrammed death-1 (PD-1) up-regulation impairs virus-specific CD8+ T-cell responses during chronic viral infection. Whether PD-1 expression influences the virus-specific CD8+ T cells in humans with acute viral infection remains largely undefined. This study aims to characterize the PD-1 expression during acute hepatitis B (AHB), and further addresses the association between the PD-1 dynamics and memory T-cell formation during acute HBV infection.

METHODSPeripheral HBV-specific CD8+ T cells from 11 HLA-A2-positive AHB patients were longitudinally quantitatively analyzed, and PD-1, memory markers CCR7, CD45RA and CD127 and activation marker CD38 on HBV-specific CD8+ T cells were measured using flow cytometric assay. Serum ALT, HBsAg, HBsAb and HBV-DNA levels were evaluated for each subject.

RESULTSAll 11 AHB patients examined had multiple pentamer-positive CD8+ T-cell responses in their early phase of HBV infection. Specifically, their PD-1 on pentamer-positive CD8+ T-cells was significantly up-regulated at the onset of their disease. Following their disease resolution, the dynamic decrease in PD-1 expression was found to correlate with the phenotypic development of memory CD8+ T cells, indicated by the increases in CCR7, CD45RA and CD127 and decrease in CD38.

CONCLUSIONPD-1-mediated negative signaling may be closely associated with memory T-cell formation during acute self-limited hepatitis B.

Acute Disease ; Adult ; Antigens, CD ; metabolism ; Apoptosis Regulatory Proteins ; metabolism ; Female ; Hepatitis B ; immunology ; metabolism ; Humans ; Immunologic Memory ; Male ; Middle Aged ; Programmed Cell Death 1 Receptor ; T-Lymphocytes, Cytotoxic ; immunology ; metabolism ; Young Adult

As a part of our ongoing search for a safe and efficient anti-tumor vaccine, we attempted to determine whether the molecular nature of certain tumor antigens would influence immune responses against tumor cells. As compared with freeze-thawed or formaldehyde-fixed tumor antigens, heat-denatured tumor antigens elicited profound anti-tumor immune responses and greatly inhibited the growth of live tumor cells. The heat-denatured tumor antigens induced a substantial increase in the anti-tumor CTL response in the absence of any adjuvant material. This response appears to be initiated by strong activation of the antigen-presenting cells, which may recognize heat-denatured protein antigens. Upon recognition of the heat-denatured tumor antigens, macrophages and dendritic cells were found to acutely upregulate the expression of co-stimulatory molecules such as B7.2, as well as the secretion of inflammatory cytokines such as IL-12 and TNF-alpha. The results of this study indicate that heat-denatured tumor extracts might elicit protective anti-tumor adaptive immune responses and also raise the possibility that a safe and efficient adjuvant-free tumor vaccine might be developed in conjunction with a dendritic cell-based tumor vaccine.
Adjuvants, Immunologic ; Animals ; Antibodies, Neoplasm/immunology ; Antibody Specificity/immunology ; Antigens, Neoplasm/immunology ; Cancer Vaccines/*immunology ; Cell Line, Tumor ; Cell Proliferation ; Cytokines/biosynthesis ; Cytotoxicity, Immunologic/immunology ; Dendritic Cells/immunology ; *Hot Temperature ; Immunity, Cellular/immunology ; Immunization ; Immunologic Memory/immunology ; Macrophages, Peritoneal/immunology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Neoplasms/*immunology/*pathology ; Survival Analysis ; T-Lymphocytes, Cytotoxic/immunology