OBJECTIVETo measure CCR5 and CXCR4 chemokine receptor expression on CD4 and CD8 T cells in HIV-1 infection and to relate levels to the distribution of CD45RO memory and CD45RA-naive subsets after effective HAART.

METHODSFour-color cytofluorometry with appropriate conjugated monoclonal antibodies (mAbs) was performed to define CD45RA and CD45RO subsets of CD4 and CD8 T cells and measure the expression of CCR5, CXCR4 in blood from 43 received HAART patients and 5 non-treated HIV and 13 healthy controls.

RESULTSThe levels of CCR5 and CXCR4 on CD4 and CD8 T cells and their CD45RO/CD45RA subsets in HIV-1-infected patients had not any statistical significance than that on control subjects and effective HAART could adjust the expression on T cells.

CONCLUSIONCXCR4/CCR5 plays an important role in the progress of HIV-1 infection. The most favorable condition for treatment should be initiated before stage B.

Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; Antiretroviral Therapy, Highly Active ; CD4-Positive T-Lymphocytes ; chemistry ; CD8-Positive T-Lymphocytes ; chemistry ; HIV-1 ; Humans ; Receptors, CCR5 ; blood ; drug effects ; Receptors, CXCR4 ; blood ; drug effects

BACKGROUNDAlthough CD4(+) T cell apoptosis and CD8(+) T cell responses have been extensively studied during HIV infection, how apoptosis signals being initiated in CD4(+) T cells still need to be elucidated. The present study was designed to characterize the function-unknown gene, C6orf120, and elucidates its primary role in tunicamycin-induced CD4(+) T apoptosis.

METHODSThe C6orf120 coding sequence was amplified from peripheral blood mononuclear cells (PBMCs) total RNA of AIDS patients. The DNA fragment was inserted into the pET-32a expression system, transformed into Escherichia coli, and preparation of C6ORF120 recombinant protein. The magnetic cell separation technology was used to prepare primary CD4(+) T cells and CD8(+) T cells. The primary T cells were cultured at 1 × 10(6) cells/ml, treated with 0, 0.1, 1, 10, 100, and 200 ng/ml of C6orf120 recombinant protein for 48 hours, then harvested for cell cycle and apoptosis analysis. Tunicamycin (0.5 µmol/L) was used to induce endoplasmic reticulum stress in Jurkat cells. The biomarker 78 KDa glucose-regulated protein (GRp78) and growth arrest and DNA damage (GADD) were used to evaluate endoplasmic reticulum stress of Jurkat cells.

RESULTSWe prepared C6ORF120 recombinant protein and its polyclonal antibody. Immunohistochemical analysis showed that C6orf120 mainly expressed in hepatocytes and cells in germinal center of lymph node. At concentration of 0.1, 1, 10, 100, and 200 ng/ml, C6orf120 recombinant protein could induce apoptosis of Jurkat cells and primary CD4(+) T cells, and promoting G2 phase of its cell cycle. Western blotting analysis showed that C6ORF120 recombinant protein increased the expression of GRp78 and GADD in Jurkat cells in vitro.

CONCLUSIONOur results suggested that C6ORF120 could induce apoptosis of CD4(+) T cells, at least in part, mediated with endoplasmic reticulum stress.

Antiviral Agents ; pharmacology ; Apoptosis ; drug effects ; Blotting, Western ; CD4-Positive T-Lymphocytes ; drug effects ; metabolism ; CD8-Positive T-Lymphocytes ; Cell Cycle ; Cells, Cultured ; Endoplasmic Reticulum Stress ; Female ; HIV Infections ; immunology ; Humans ; Immunohistochemistry ; Male ; Microscopy, Confocal ; Proteins ; genetics ; metabolism ; Tunicamycin ; pharmacology

OBJECTIVETo observe effect of oral scorpio and scolopendra powder on T-cell subsets in peripheral blood and intestine from rats with collagen induced arthritis (CIA).

METHOD60 rats were randomly divided into 6 groups: normal control group, model control group, low-dose scorpio and scolopendra group, middle-dose scorpio and scolopendra group, high-dose scorpio and scolopendra group, and type II collagen group. Rat's rheumatoid arthritis was induced by collagen II (C II). Level of T-cell subsets from peripheral blood and intestine was measured by flow cytometry.

RESULTCD4+ T cellular level was obviously increased (P < 0.05 or P < 0.01) or kept increased tendency in peripheral blood and intestine from the model group compared with that of the normal group, while the ratio of CD4+/CD8+ in intestine was obviously descent but the contrary in peripheral blood (P < 0.05 or P < 0.01). CD4+, CD8+ T cellular level in intestine were obviously descent and the ratio of CD4+ /CD8+ increased in all treated groups when compared with in the model group (P < 0.05 or P < 0.01). However, CD4+ T cellular level and the ratio of CD4+/CD8+ in peripheral blood were remarkablely decreased.

CONCLUSIONThe mechanism that scorpio and scolopendra could treat rat's rheumatoid arthritis may be regulating balance of T-lymphocyte subsets in peripheral blood and intestine.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Arthritis, Experimental ; immunology ; Arthritis, Rheumatoid ; immunology ; CD4 Lymphocyte Count ; CD4-Positive T-Lymphocytes ; CD8-Positive T-Lymphocytes ; Female ; Intestinal Mucosa ; immunology ; Medicine, Chinese Traditional ; Rats ; Rats, Wistar ; Scorpions ; chemistry ; T-Lymphocyte Subsets ; drug effects ; immunology

We previously demonstrated that stimulation of bovine peripheral blood mononuclear cells (PBMCs) with staphylococcal enterotoxin C (SEC), led to an inversion of the CD4(+):CD8(+) T cell ratio and generation of an atypical CD8(+) T cell subpopulation expressing CD26. In the present study, we examined T cell apoptosis and proliferation profiles of PBMC subpopulations in cultures stimulated with SEC. Unlike when stimulated with concanavalin A, nucleic acid synthesis in bovine PBMC cultures stimulated with SEC was low during the first four days but increased greatly on day 5. In contrast, nucleic acid synthesis in human PBMC cultures stimulated with SEC increased continuously. To investigate the mechanism of delayed bovine T cell proliferation, various cell phenotypes were monitored. The inversion of the bovine CD4(+):CD8(+) T cell ratio in PBMC cultures stimulated by SEC was associated with higher proliferation and lower apoptosis of CD8(+) T cells compared to CD4(+) T cells. The mRNA levels for interleukin (IL)-4 and IL-13 were sustained over 4 days but IL-12 mRNA levels dropped to background on day 2. These data suggest that SEC induces a prolonged Th-2- biased microenvironment, and together with the inversion of the bovine CD4(+):CD8(+) T cell ratios in bovine PBMC cultures with SEC, may in part explain the inability of the mammary immune system to establish an effective response to Staphylococcus aureus infections.
Animals ; Apoptosis/drug effects/immunology ; CD4-CD8 Ratio/veterinary ; CD4-Positive T-Lymphocytes/drug effects/*immunology/microbiology ; CD8-Positive T-Lymphocytes/drug effects/*immunology/microbiology ; Cattle ; Concanavalin A/pharmacology ; Cytokines/genetics/immunology ; Enterotoxins/*pharmacology ; Female ; Lymphocyte Activation/drug effects ; Mastitis, Bovine/immunology/*microbiology ; Reverse Transcriptase Polymerase Chain Reaction/veterinary ; Staphylococcal Infections/immunology/microbiology/*veterinary ; Staphylococcus aureus/*immunology

BACKGROUNDThe prospects of using immature CD8a(+) dendritic cells (DC2) to establish transplant immunologic tolerance and treatments for autoimmune diseases in the future are promising. However, the methods for inducing DC2 are still being explored. The present study was aimed to investigate the optimal in vitro conditions for preparing large numbers of predominant DC2 from murine bone marrow cells.

METHODSThree groups of bone marrow cells cultured under different conditions were examined, namely a cytokine-induced experimental group (cytokine group), a control group with a low concentration of granulocyte-macrophage colony stimulating factor (GM-CSF, low GM-CSF group) and a control group without endogenous cytokines. The cytokine group was cultured with 5 ng/ml GM-CSF, 25 ng/ml Flt3 ligand (Flt3L), 20 ng/ml interleukin 4 (IL-4) and 100 ng/ml stem cell factor (SCF). The low GM-CSF control group was cultured with 0.4 ng/ml GM-CSF, 25 ng/ml Flt3L and 100 ng/ml SCF, without IL-4. The control group without exogenous cytokines was cultured without additional cytokines. All cells were cultured at 37 degrees C under 5% CO2. On days 3, 7 and 16, 4-color flow cytometry was carried out to analyze the cell phenotypes, and the total cell numbers were counted to analyze the cell yields. Phase-contrast microscopy was used to observe the cell morphologies.

RESULTSThe cytokine group exhibited higher proportions of typical immature CD8a(+) DC, especially on day 3, but the total cell number and DC2 proportion decreased during prolonged culture. The low GM-CSF control group showed the same tendencies as the cytokine group on days 16 and 22, but produced higher total cell numbers (P<0.05) with lower DC2 proportions and cell numbers. The control group without exogenous cytokines spontaneously generated a certain proportion of DC2, but with low total cell and DC2 numbers that decreased rapidly, especially during prolonged culture (days 7 and 16, P<0.05).

CONCLUSIONSCulture in the presence of 5 ng/ml GM-CSF, 25 ng/ml Flt3L, 20 ng/ml IL-4 and 100 ng/ml SCF can rapidly induce large quantities of predominant immature CD8a(+) DC from murine bone marrow cells. Therefore, these represent optimal culture conditions for preparing murine immature DC2 in vitro.

Animals ; Bone Marrow Cells ; cytology ; drug effects ; CD8 Antigens ; metabolism ; CD8-Positive T-Lymphocytes ; cytology ; drug effects ; Cell Culture Techniques ; methods ; Cells, Cultured ; Flow Cytometry ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Male ; Mice ; Mice, Inbred BALB C ; Microscopy, Phase-Contrast

BACKGROUNDSjögren syndrome (SS) is a systematic autoimmune disease, on which traditional therapeutic agents show limited effect. More effective agents with longer-lasting and fewer side effects are needed in the clinic. The aim of this study was to investigate the effects of Ganoderma lucindum spores (GLS) on sialoadenitis of nonobese diabetic (NOD) mice.

METHODSThirty-two female NOD mice were assigned randomly into 4 groups: low-dose GLS-treated (L-GLS) group and high-dose GLS-treated (H-GLS) group, a dexamethasone group, and a normal saline (NS) control group. Stimulated total saliva flow rate (STFR), area of lymphocytic infiltration in submandibular glands and ratios of CD4(+) and CD8(+) T lymphocytes and B lymphocytes in peripheral blood as well as apoptosis of these subsets and serum IgG level were tested after 10 weeks of treatments. Differences among the groups were analyzed by one-way analysis of variance (ANOVA), Student-Newman-Keuls Test (SNK) was used between each two groups and a P < 0.05 was considered statistically significant.

RESULTSSTFR of the high-dose GLS group increased significantly after a 10-week treatment compared with those of the NS control group (P < 0.05). The incidence of sialoadenitis in GLS-treated NOD mice groups showed no significant difference compared with the control group (P > 0.05), but the area of lymphocytic foci in both the H-GLS and L-GLS groups decreased significantly to 50% of the NS control group (P < 0.05); the ratio of CD4(+)/CD8(+) T lymphocytes and apoptosis of B lymphocytes of NOD mice with sialoadenitis were less and apoptosis of CD4(+) and CD8(+) T lymphocytes were significantly increased compared with the control group (P < 0.05). After pretreatment with H-GLS before sialoadenitis onset, the ratio of CD4(+)/CD8(+) T lymphocyte and the serum IgG levels of NOD mice decreased significantly (P < 0.05).

CONCLUSIONSPretreatment with H-GLS can relieve symptoms of sialoadenitis in NOD mice. GLS has some protective effects on sialoadenitis in NOD mice through increasing STFR and decreasing the area of lymphocytic foci by regulating the ratio of CD4(+)/CD8(+) T and apoptosis of B lymphocytes.

Animals ; Apoptosis ; drug effects ; B-Lymphocytes ; cytology ; drug effects ; CD4-Positive T-Lymphocytes ; cytology ; drug effects ; CD8-Positive T-Lymphocytes ; cytology ; drug effects ; Drugs, Chinese Herbal ; chemistry ; therapeutic use ; Female ; Immunoglobulin G ; blood ; Lymphocyte Subsets ; cytology ; drug effects ; Mice ; Mice, Inbred NOD ; Random Allocation ; Reishi ; chemistry ; Sjogren's Syndrome ; blood ; drug therapy ; Spores, Fungal ; chemistry

Anti-Bacterial Agents ; therapeutic use ; CD4-CD8 Ratio ; CD4-Positive T-Lymphocytes ; cytology ; drug effects ; CD8-Positive T-Lymphocytes ; cytology ; drug effects ; Child ; Child, Preschool ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Infant ; Male ; Phytotherapy ; Pneumonia ; drug therapy ; Powders ; Recurrence ; Respiratory Tract Infections ; drug therapy ; Treatment Outcome

The study was purposed to explore the changes of CD4(+)CD25(+) T regulatory cells in patients with multiple myeloma before and (MM) after treatment with thalidomide so as to provide evidences for effective immunotherapy. The population of CD3(+) T, CD4(+) T, CD8(+) T, NK and CD4(+)CD25(+) Treg in patients with MM were detected by flow cytometry. Statistical significance of differences in different groups was determined by using the t test. A p value of less than 0.05 was considered to be significant. The results showed that the percentage of CD4(+)CD25(+ high) T in patients with MM was significantly higher than that of the healthy donors (p > 0.01). The population of CD4(+)CD25(+ high) Treg cells in patients with response to thalidomide was significantly decreased (p < 0.01), but the population of these cells in patients without response not changed significantly (p > 0.05), as compared with patients before treatment. In 16 patients who achieved complete remission after chemotherapy, the population of CD4(+)CD25(+ high) T was 6.91 +/- 1.12%, which was slightly higher than that before treatment. The population of CD3(+) T, CD4(+) T, CD8(+) T, NK and CD4(+)CD25(+) Treg significantly increased in patients with positive response to thalidomide, but the population of CD8(+) T remained unchanged. It is concluded that the significant increase of CD4(+)CD25(+) regulatory T cells in peripheral blood of patients with MM is concerned with the MM pathogenesis; thalidomide may exert its anti-MM effects by down-regulating CD4(+)CD25(+) Treg.
Adult ; Aged ; Aged, 80 and over ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Female ; Humans ; Immunosuppressive Agents ; therapeutic use ; Killer Cells, Natural ; immunology ; Male ; Middle Aged ; Multiple Myeloma ; drug therapy ; immunology ; T-Lymphocyte Subsets ; immunology ; T-Lymphocytes, Regulatory ; drug effects ; immunology ; Thalidomide ; therapeutic use

BACKGROUNDArthritogenic T lymphocytes with common T cell receptor (TCR) Vbeta clonotypes, infiltrating in the articulars of rheumatoid arthritis (RA) patients, play a central role in the pathogenesis of RA. TCR Vbeta5.2 and TCR Vbeta8.2 are the main pathogenic T cell clonotypes in the course of collagen-induced arthritis (CIA) progression in Lewis rats. To investigate a TCR-based immunotherapy for RA, we constructed recombinant DNA vaccines encoding TCR Vbeta5.2 and TCR Vbeta8.2, and evaluated the inhibitive effects of the two vaccines on CIA rats.

METHODSGenes encoding TCR Vbeta5.2 and TCR Vbeta8.2 were amplified by RT-PCR from spleen lymphocytes of Lewis rats and cloned into the eukaryotic expression vector pTargeT. The expression of vaccines was confirmed by RT-PCR and immunohistochemistry. The inhibitive effects of the vaccines on articulars of CIA rats were assessed with arthritis index evaluation and histology. Interferon gamma (IFN-gamma) and interleukin (IL)-4 production by spleen lymphocytes were tested with enzyme-linked immunospot assay (ELISPOT) technique, the changes in peripheral CD4(+) and CD8(+) lymphocyte populations were tested by flow cytometry, and the level of anti-CII antibody in serum was assayed by enzyme-linked immunosorbent assay (ELISA).

RESULTSRecombinant DNA vaccines pTargeT-TCR Vbeta5.2 and pTargeT-pTCR Vbeta8.2 were successfully constructed. Both vaccines inhibited CIA, which alleviated the arthritis index score (P < 0.05), decreased the level of IFN-gamma (P < 0.05), and reduced the ratio of CD4(+)/CD8(+) lymphocytes (P < 0.05) and the anti-CII antibody in serum (P < 0.05). In addition, the histological change in DNA-vaccinated rats was less serious than CIA rats. Compared to pTCR Vbeta 8.2 and pTCR Vbeta 5.2 groups, the group that was injected with a combination of the two vaccines showed stronger inhibitive effects on CIA than either individual vaccine.

CONCLUSIONThe recombinant plasmids pTargeT-TCR Vbeta5.2 and pTargeT-TCR Vbeta8.2 have obvious inhibatory effects on CIA rats and better effects could be achieved when the vaccines were used in combination.

Animals ; Arthritis, Experimental ; metabolism ; prevention & control ; CD4-Positive T-Lymphocytes ; drug effects ; CD8-Positive T-Lymphocytes ; drug effects ; Enzyme-Linked Immunosorbent Assay ; Female ; Flow Cytometry ; Immunohistochemistry ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism ; Muscles ; drug effects ; metabolism ; Peptide Fragments ; antagonists & inhibitors ; Rats ; Rats, Inbred Lew ; Receptors, Antigen, T-Cell, alpha-beta ; antagonists & inhibitors ; Reverse Transcriptase Polymerase Chain Reaction ; Vaccines, DNA ; pharmacology

BACKGROUNDAmong HIV-infected patients initiating antiretroviral therapy (ART), early changes in CD4+ T-cell subsets are well described. However, HIV-infected late presenters initiating treatment present with a suboptimal CD4+ T-cell reconstitution and remain at a higher risk for AIDS and non-AIDS events. Therefore, factors associated with CD4+ T-cell reconstitution need to be determined in this population, which will allow designing effective immunotherapeutic strategies.

METHODSThirty-one adult patients with baseline CD4+ T-cell count <350 cells/mm3 exhibiting viral suppression after ART initiation were followed in the HIV/AIDS research center of Peking Union Medical College Hospital in Beijing, China, from October 2002 to September 2013. Changes in T-cell subsets and associated determinants were measured.

RESULTSMedian baseline CD4+ T-cell count was 70 cells/mm3. We found a biphasic reconstitution of T-cell subsets and immune activation: a rapid change during the first 6 months followed by a more gradual change over the subsequent 8 years. Baseline CD4+ T-cell count >200 cells/mm3 in comparison to CD4+ T-cell count ≤200 cells/mm3 was associated with more complete immune Reconstitution (77.8% vs. 27.3% respectively; P = 0.017) and normalized CD4/CD8 ratio. We showed that the baseline percentage of naive CD4+ T-cell was a predictive marker for complete immune reconstitution (area under receiver operating characteristic curve 0.907), and 12.4% as cutoff value had a sensitivity of 84.6% and a specificity of 88.2%.

CONCLUSIONSBaseline naive CD4+ T-cell percentage may serve as a predictive marker for optimal immune reconstitution during long-term therapy. Such study findings suggest that increasing thymic output should represent an avenue to improve patients who are diagnosed late in the course of infection.

Adult ; Antiretroviral Therapy, Highly Active ; methods ; CD4 Lymphocyte Count ; CD4-CD8 Ratio ; CD4-Positive T-Lymphocytes ; metabolism ; Female ; HIV Infections ; drug therapy ; immunology ; metabolism ; HIV-1 ; drug effects ; immunology ; pathogenicity ; Humans ; Male ; Prospective Studies ; T-Lymphocyte Subsets ; immunology