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1.
Article in Chinese | WPRIM | ID: wpr-286883

ABSTRACT

<p><b>OBJECTIVE</b>To analyze the characteristics of lymphocyte phenotypes in hepatitis B virus (HBV) transgenic mice and the effect of exogenous interferon-α on virological profiles and lymphocytes phenotypes of the mice.</p><p><b>METHODS</b>HBV transgenic mice and wild-type (WT) mice were examined for serum levels of HBsAg, HBcAb, IL-21, and IL-6 using ELISA. The frequencies of CD4(+)T and CD19(+)B cells separated from the liver, spleen, and peripheral blood were detected by flow cytometry. Nine HBV transgenic mice were injected subcutaneously with recombinant mouse interferon alpha (rmIFN-α) and another 9 transgenic mice were injected with PBS, and their HBsAg, HBV DNA, IL-6, and IL-21 levels and frequencies of peripheral blood CD4(+)T and CD19(+)B cells were detected.</p><p><b>RESULTS</b>HBV transgenic mice showed a high level of HBsAg with a detectable level of HBcAb and significantly increased serum levels of IL-21 and IL-6 as compared with WT mice (P<0.05). The transgenic mice had a significantly lower frequency of CD4(+) T cells in the peripheral blood, liver and spleen (P<0.05) but a significantly higher frequency of CD19(+) B cells in the liver (P<0.05). An inverse correlation between intrahepatic CD4(+) T cell frequency and serum HBsAg level while a positive correlation between intrahepatic CD19(+) B cell frequency and HBcAb level were found in HBV transgenic mice. Administration of rmIFN-α significantly increased the frequencies of CD4(+) T and CD19(+) B cells in the peripheral blood and the serum level of IL-6 in HBV transgenic mice (P<0.05).</p><p><b>CONCLUSION</b>HBV transgenic mice have lymphocyte subset dysregulation and exogenous interferon-α can modulate the immune function of the mice by regulating the frequencies of lymphocyte subsets.</p>


Subject(s)
Animals , Antiviral Agents , Pharmacology , B-Lymphocytes , DNA, Viral , Blood , Hepatitis B , Drug Therapy , Allergy and Immunology , Hepatitis B Antibodies , Blood , Hepatitis B Surface Antigens , Blood , Hepatitis B virus , Interferon-alpha , Pharmacology , Interleukin-6 , Blood , Interleukins , Blood , Liver , Allergy and Immunology , Lymphocyte Subsets , Cell Biology , Mice , Mice, Transgenic , Phenotype , T-Lymphocytes
2.
Braz. j. med. biol. res ; 48(12): 1095-1100, Dec. 2015. graf
Article in English | LILACS | ID: lil-762920

ABSTRACT

In DNA vaccines, the gene of interest is cloned into a bacterial plasmid that is engineered to induce protein production for long periods in eukaryotic cells. Previous research has shown that the intramuscular immunization of BALB/c mice with a naked plasmid DNA fragment encoding the Mycobacterium leprae 65-kDa heat-shock protein (pcDNA3-Hsp65) induces protection against M. tuberculosis challenge. A key stage in the protective immune response after immunization is the generation of memory T cells. Previously, we have shown that B cells capture plasmid DNA-Hsp65 and thereby modulate the formation of CD8+ memory T cells after M. tuberculosis challenge in mice. Therefore, clarifying how B cells act as part of the protective immune response after DNA immunization is important for the development of more-effective vaccines. The aim of this study was to investigate the mechanisms by which B cells modulate memory T cells after DNA-Hsp65 immunization. C57BL/6 and BKO mice were injected three times, at 15-day intervals, with 100 µg naked pcDNA-Hsp65 per mouse. Thirty days after immunization, the percentages of effector memory T (TEM) cells (CD4+ and CD8+/CD44high/CD62Llow) and memory CD8+ T cells (CD8+/CD44high/CD62Llow/CD127+) were measured with flow cytometry. Interferon γ, interleukin 12 (IL-12), and IL-10 mRNAs were also quantified in whole spleen cells and purified B cells (CD43−) with real-time qPCR. Our data suggest that a B-cell subpopulation expressing IL-10 downregulated proinflammatory cytokine expression in the spleen, increasing the survival of CD4+ TEM cells and CD8+ TEM/CD127+ cells.


Subject(s)
Animals , Male , Mice , B-Lymphocytes/immunology , Heat-Shock Proteins/immunology , Immunomodulation/genetics , /genetics , RNA, Messenger/immunology , T-Lymphocyte Subsets/immunology , B-Lymphocytes/metabolism , Flow Cytometry , Gene Expression/genetics , Heat-Shock Proteins/therapeutic use , Immunologic Memory/physiology , Immunophenotyping/classification , Inflammation Mediators/analysis , Interferon-gamma/analysis , /immunology , /analysis , Mice, Knockout , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , RNA, Messenger/genetics , Spleen/cytology , Spleen/immunology , T-Lymphocyte Subsets/classification , Vaccines, DNA/immunology , Vaccines, DNA/therapeutic use
3.
Chinese Journal of Stomatology ; (12): 585-589, 2015.
Article in Chinese | WPRIM | ID: wpr-294669

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the possible role and significance of soluble programmed death-1 (sPD-1)/soluble programmed death ligand 1 (sPD-L1) in the pathogenesis of oral lichen planus (OLP).</p><p><b>METHODS</b>Thirty-six patients with OLP (20 cases of reticular OLP and 16 cases of erosive OLP) were enrolled in this study, and 18 healthy people served as controls. Lymphocyte subsets (CD3⁺, CD4⁺, CD8⁺, CD19⁺, CD16⁺ + 56⁺) were examined by flow cytometric analysis and humoral immunity indexes (IgG, IgA, IgM, C3, C4) tested by nephelometry immunoassay. The levels of sPD-1 and sPD-L1 proteins in serum of patients with OLP were determined by enzyme-linked immunosorbent assay. The correlations between the level of sPD-1, sPD-L1 proteins and the immune status and clinical characteristics of patients with OLP were analyzed by SPSS 19.0.</p><p><b>RESULTS</b>CD3⁺, CD4⁺, CD8⁺, CD16⁺ + 56⁺ in patients with OLP were decreased compared with the normal value, while CD19⁺ in patients with OLP was increased compared with the normal value (P < 0.05). C3 and C4 in patients with OLP were decreased compared with the normal value, but IgM in patients with OLP was increased (P < 0.05). The levels of sPD-1 and sPD-L1 proteins in patients with OLP were significantly higher than that in control group [26.10(8.81, 40.00) ng/L vs 17.65(0.00, 26.10) ng/L, 29.53(21.47, 36.76) ng/L vs 22.79(1.19, 28.29) ng/L] (P < 0.05), but the expression of sPD-1 and sPD-L1 was not related with clinical characteristics of OLP. There were negative correlations between the levels of sPD-1 protein and CD4⁺ T cells or CD16⁺ + 56⁺ cells (r1 = -0.378, P1 = 0.007; r2 = -0.365, P2 = 0.009), while there was a positive correlation between the levels of sPD-1 and CD19⁺ B cells (r = 0.482, P = 0.000). There was a negative correlation between sPD-L1 expression level and CD4⁺ and a positive correlation between sPD-L1 expression level and IgG (r¹ = -0.286, P1 = 0.044; r² = 0.365, P₂= 0.029).</p><p><b>CONCLUSIONS</b>In patients with OLP, the cellular immune function is low with humoral immunity function disorder. PD-1/PD-L1 signaling pathway, which might be influenced by the involvement of sPD-1 and sPD-L1 proteins in a certain extent, may play an important role in the immune pathogenesis of OLP.</p>


Subject(s)
B-Lymphocytes , B7-H1 Antigen , Blood , Case-Control Studies , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Humans , Immunoglobulins , Blood , Lichen Planus, Oral , Blood , Allergy and Immunology , Nephelometry and Turbidimetry , Methods , Programmed Cell Death 1 Receptor , Blood , Signal Transduction , T-Lymphocytes
4.
Chinese Journal of Oncology ; (12): 497-500, 2015.
Article in Chinese | WPRIM | ID: wpr-286792

ABSTRACT

<p><b>OBJECTIVE</b>To explore the effects of radiofrequency ablation(RFA) on immune system and lung metastasis in a mouse model of triple negative breast cancer 4T1.</p><p><b>METHODS</b>Mouse breast cancer 4T1 cells were injected into the right hind limb of female Bal B/c mice. When the tumor size was 6-8 mm in diameter, RFA was used to treat the transplanted breast cancer in mice. We examined the splenic lymphocyte subsets by flow cytometry at different time points after RFA. Fourteen days after treatment, we sacrificed the mice of both control and treatment groups, counted the number of lung metastatic nodules, and detected the changes of splenic lymphocyte subsets by flow cytometry.</p><p><b>RESULTS</b>RFA basically eliminated the orthotopic carcinoma with a low local recurrence rate. After the RFA treatment, the amount of spleic CD4⁺ T cells, CD8⁺ T cells, B cells, NK and NKT cells was increased. Fourteen days after the RFA treatment, all mice were sacrificed, and the lung metastatic nodules were 24 ± 18 in the control group and 81 ± 35 in the RFA-treated group (P = 0.012). The mechanism of suppression of metastatic lung cancers was related to the increase of splenic CD4⁺ T cells, CD8⁺ T cells, B cells and NK cells, and the decrease of myeloid-derived suppressor cells.</p><p><b>CONCLUSIONS</b>RFA can enhance the anti-tumor immunity and effectively inhibit lung metastasis of 4T1 cell-induced breast cancer, and has a good potential effect in the treatment of triple-negative breast cancer and the control of distant metastasis.</p>


Subject(s)
Animals , B-Lymphocytes , Cell Biology , CD4-Positive T-Lymphocytes , Cell Biology , CD8-Positive T-Lymphocytes , Cell Biology , Catheter Ablation , Female , Flow Cytometry , Humans , Killer Cells, Natural , Cell Biology , Lung Neoplasms , Allergy and Immunology , Mice , Mice, Inbred BALB C , Neoplasm Recurrence, Local , Triple Negative Breast Neoplasms , Allergy and Immunology , Pathology , General Surgery , Tumor Burden
5.
Article in Chinese | WPRIM | ID: wpr-747178

ABSTRACT

OBJECTIVE@#To investigate the B lymphocytes(CD20) and T-lymphocyte subsets (CD4, CD8) expression and clinical significance in chronic rhinosinusitis.@*METHOD@#Immunohistochemical technique was applied to detect the expression of the B lymphocytes (CD20), helper T lymphocytes (CD4) and the cytotoxic T lymphocyte (CD8) in 15 patients with chronic rhinosinusitis without nasal polyps group (CRSsNP). 12 patients of chronic nasal sinusitis with nasal polyp (CRSwNP), 7 cases of recurrent CRSwNP group and 13 patients with inferior turbinate mucosa in the control group. The Mann-Whitney U test was used to analyze CD)20, CD4 and CD8 between the experimental groups and the control group. One- Way ANOVA was used to compare the lymphocyte infiltration between the experiment groups.@*RESULT@#Patients with CRS had higher infiltration of the B lymphocytes (CD20) and T lymphocyte subsets (CD4, CD88) than the patients of the control group (P < 0.05). The expression of T lymphocyte subsets (CD4, CD8) in CRSwNP and recurrent CRSwNP groups was significantly higher than in CRSsNP (P < 0.05). The expression of CD4 in the recurrent CRSwNP group was obviously higher than that in CRSsNP and CRSwNP (P < 0.01).@*CONCLUSION@#The B lymphocytes, helper T cells and cytotoxic T cells have the high expression in CRS groups, which involved with the formation of inflammation. Furthermore nasal polyps are closely related to T lymphocyte infiltration.


Subject(s)
Adolescent , Adult , B-Lymphocytes , Cell Biology , Case-Control Studies , Child , Chronic Disease , Female , Humans , Male , Middle Aged , Nasal Polyps , Rhinitis , Allergy and Immunology , Sinusitis , Allergy and Immunology , T-Lymphocyte Subsets , Cell Biology , Young Adult
6.
Chinese Journal of Hematology ; (12): 1028-1032, 2012.
Article in Chinese | WPRIM | ID: wpr-323498

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the number of peripheral blood CD5(+) B cells and their ability of secreting IL-10 in patients with immune thrombocytopenia (ITP).</p><p><b>METHODS</b>Peripheral blood lymphocytes were isolated from 57 pre-treated, 40 post-treated ITP patients and 25 controls using Ficoll-Hypaque density centrifugation and then stained with PE-CD5/FITC-CD19 for flow cytometric analysis. After 24-hour culture, lymphocytes were stained with APC-IL-10 for intracellular cytokine detection. ELISA assay was employed to determine IL-10 concentration in supernatants.</p><p><b>RESULTS</b>The percentage and absolute number of CD5(+) B cells in peripheral blood from pre-treated ITP patients were significantly higher than that from normal controls (3.75 ± 2.37)% vs (2.10 ± 1.08)%, P < 0.01; (6.29 ± 5.77)× 10(7)/L vs (3.06 ± 1.90)× 10(7)/L, P < 0.01. CD5(+) B cells expressed more intracellular IL-10 than other lymphocyte subsets both in ITP patients and normal controls. The percentages of IL-10(+) cells within CD5(+) B cells in pre-treated ITP patients and normal controls were (29.51 ± 20.73)% and(15.90 ± 9.58)%, respectively(P < 0.01). Intracellular mean fluorescence intensity (MFI) of IL-10 in CD5(+) B cells was 27.95 ± 13.99 in pre-treated patients, which was significantly higher than that in controls (P < 0.01). In contrast, IL-10 concentration in supernatants was (173.05 ± 102.50) ng/L in pre-treated ITP group, which was lower than that (230.61 ± 76.96) ng/L in controls. In patients who achieved remission, the number of CD5(+) B cells decreased to level comparable to normal controls. While intracellular IL-10 MFI of CD5(+) B cells in post-treated ITP patients remained as high as in pre-treated ones, the IL-10 concentration in supernatants increased to level similar to controls.</p><p><b>CONCLUSION</b>The significantly increased number of CD5(+) B cells and accumulated IL-10 in CD5(+) B cells suggested impaired IL-10 secretion in ITP patients. The number and the ability of secreting IL-10 of CD5(+) B cells could be restored after effective treatments in patients with ITP.</p>


Subject(s)
Adult , Aged , B-Lymphocytes , Allergy and Immunology , Metabolism , CD5 Antigens , Metabolism , Case-Control Studies , Female , Humans , Interleukin-10 , Blood , Male , Middle Aged , Purpura, Thrombocytopenic, Idiopathic , Blood , Allergy and Immunology , Young Adult
7.
Protein & Cell ; (12): 571-580, 2012.
Article in English | WPRIM | ID: wpr-757257

ABSTRACT

Epithelial tissues covering the external and internal surface of a body are constantly under physical, chemical or biological assaults. To protect the epithelial tissues and maintain their homeostasis, multiple layers of immune defense mechanisms are required. Besides the epithelial tissue-resident immune cells that provide the first line of defense, circulating immune cells are also recruited into the local tissues in response to challenges. Chemokines and chemokine receptors regulate tissue-specific migration, maintenance and functions of immune cells. Among them, chemokine receptor CCR10 and its ligands chemokines CCL27 and CCL28 are uniquely involved in the epithelial immunity. CCL27 is expressed predominantly in the skin by keratinocytes while CCL28 is expressed by epithelial cells of various mucosal tissues. CCR10 is expressed by various subsets of innate-like T cells that are programmed to localize to the skin during their developmental processes in the thymus. Circulating T cells might be imprinted by skin-associated antigen- presenting cells to express CCR10 for their recruitment to the skin during the local immune response. On the other hand, IgA antibody-producing B cells generated in mucosa-associated lymphoid tissues express CCR10 for their migration and maintenance at mucosal sites. Increasing evidence also found that CCR10/ligands are involved in regulation of other immune cells in epithelial immunity and are frequently exploited by epithelium-localizing or -originated cancer cells for their survival, proliferation and evasion from immune surveillance. Herein, we review current knowledge on roles of CCR10/ligands in regulation of epithelial immunity and diseases and speculate on related important questions worth further investigation.


Subject(s)
B-Lymphocytes , Cell Biology , Allergy and Immunology , Cell Lineage , Cell Movement , Genetics , Allergy and Immunology , Chemokine CCL27 , Genetics , Allergy and Immunology , Chemokines, CC , Genetics , Allergy and Immunology , Epithelial Cells , Cell Biology , Allergy and Immunology , Epithelium , Allergy and Immunology , Gene Expression Regulation , Allergy and Immunology , Humans , Immunity, Mucosal , Immunoglobulin A , Allergy and Immunology , Mucous Membrane , Cell Biology , Allergy and Immunology , Receptors, CCR10 , Genetics , Allergy and Immunology , Signal Transduction , Genetics , Allergy and Immunology , T-Lymphocytes , Cell Biology , Allergy and Immunology
8.
Chinese Medical Journal ; (24): 275-280, 2012.
Article in English | WPRIM | ID: wpr-333502

ABSTRACT

<p><b>BACKGROUND</b>Very few researchers have studied the changes in peripheral lymphocyte patterns in adult tuberculosis (TB) and even less researches have been conducted in pediatric TB. In this study, we obtained blood samples from 114 Chinese pediatric TB patients and 116 matched controls to study the association of phenotypic subsets of peripheral lymphocytes with different clinical phenotypes of TB.</p><p><b>METHODS</b>The subjects were classified as the control group and the TB patients group which were further divided into a pulmonary TB group and an extra-pulmonary TB group (more serious than the former). The distribution of lymphocyte subpopulations, including T lymphocytes, CD4(+) T lymphocytes, CD8(+) T lymphocytes, B lymphocytes, and natural killer (NK) cells, were quantitatively analyzed by flow cytometry.</p><p><b>RESULTS</b>Compared to the healthy controls, TB infection was associated with significantly higher B cell (P < 0.0001), and lower T cell (P = 0.029) and NK cell (P < 0.0001) percentages. Compared to pulmonary TB patients, extra-pulmonary TB was associated with relatively higher B cell (P = 0.073), and lower T cell percentages (P = 0.021), higher purified protein derivative (PPD) negative rate (P = 0.061), and poorer PPD response (P = 0.010). Most pulmonary TB cases were primary pulmonary TB (89.1%), and most extra-pulmonary TB cases had TB meningitis (72.1%).</p><p><b>CONCLUSIONS</b>This study demonstrates changes in the lymhocyte distribution in children suffering from different clinical phenotypes of TB; such as primary pulmonary TB, and TB meningitis. These patterns may have significance in understanding the pathogenesis and prognostic markers of the disease, and for developing immunomodulatory modalities of therapy.</p>


Subject(s)
Adolescent , Asian Continental Ancestry Group , B-Lymphocytes , Allergy and Immunology , CD4-Positive T-Lymphocytes , Allergy and Immunology , CD8-Positive T-Lymphocytes , Allergy and Immunology , Child , Child, Preschool , Female , Flow Cytometry , Humans , Immunophenotyping , Methods , Infant , Killer Cells, Natural , Allergy and Immunology , Lymphocytes , Allergy and Immunology , Male , T-Lymphocytes , Allergy and Immunology , Tuberculosis , Allergy and Immunology
9.
Article in English | WPRIM | ID: wpr-192556

ABSTRACT

Most of the previous studies on immune dysregulation in end-stage renal disease (ESRD) have focused on T cell immunity. We investigated B cell subpopulations in ESRD patients and the effect of hemodialysis (HD) on B cell-associated immune profiles in these patients. Forty-four ESRD [maintenance HD patients (n = 27) and pre-dialysis patients (n = 17)] and 27 healthy volunteers were included in this study. We determined the percentage of B cell subtypes, such as mature and immature B cells, memory B cells, and interleukin (IL)-10+ cells, as well as B cell-producing cytokines (IL-10, IL-4 and IL-21) by florescent activated cell sorting (FACS). B cell-associated gene expression was examined using real-time PCR and B cell producing cytokines (IL-10, IL-4 and IL-21) were determined using an enzyme-linked immunosorbent assay (ELISA). The percentage of total B cells and mature B cells did not differ significantly among the three groups. The percentages of memory B cells were significantly higher in the pre-dialysis group than in the HD group (P 0.05) between the two subgroups within the ESRD group, but the serum IL-10 concentration was significantly lower in the pre-dialysis group (P < 0.01). The results of this study demonstrate significantly altered B cell-associated immunity. Specifically, an imbalance of immature and memory B cells in ESRD patients was observed, with this finding predominating in pre-dialysis patients.


Subject(s)
Adaptor Proteins, Signal Transducing/genetics , Adult , Antigens, CD19/metabolism , B-Lymphocyte Subsets/immunology , B-Lymphocytes/immunology , Cytokines/biosynthesis , Female , Humans , Immunophenotyping , Interleukin-10/metabolism , Kidney Failure, Chronic/immunology , Leukocytes, Mononuclear/metabolism , Male , Middle Aged , Proto-Oncogene Proteins/genetics , T-Lymphocytes, Regulatory/immunology
10.
Chinese Medical Journal ; (24): 1517-1523, 2011.
Article in English | WPRIM | ID: wpr-353952

ABSTRACT

<p><b>BACKGROUND</b>The cause of late-onset hemorrhagic cystitis (LOHC) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) remains obscure. In clinical practice, some LOHC patients respond to immunosuppression. The aim of this study was to determine the immune pathogenesis of LOHC post allo-HSCT.</p><p><b>METHODS</b>With the diagnosis of LOHC, patients were given initial treatment consisting of fluid hydration, alkalization and forced diuresis, and empirical anti-viral therapy for 10 - 14 days or until a week after the virus became negative. The nonresponders were applied corticosteroid. Seven to ten days later, patients' response was evaluated. Along with treatment, CD19(+) B lymphocyte subsets were measured at various study points.</p><p><b>RESULTS</b>From October 2009 to March 2010, we found 28 cases of LOHC occurred in 25 patients who underwent allo-HSCT in our hospital. Except that three cases were not treated according to the protocol, the other 25 cases were divided into three groups: anti-virus responders (Group A, n = 6), corticosteroid responders (Group B1, n = 16), corticosteroid and anti-virus nonresponders (Group C, n = 3) according to their clinical response. Percentages of CD19(+)CD5(+) B lymphocytes were not significantly different among three groups at onset of LOCH. However, in Group B1 after the first anti-virus phase, percentages of CD19(+)CD5(+) lymphocytes significantly increased comparing with those at onset (P = 0.022), and then significantly decreased at PR (P = 0.003) and CR (P = 0.002) with corticosteroid treatment. But significant change was not observed in Groups A and C.</p><p><b>CONCLUSION</b>The immune etiology seems to be involved in the development of LOHC and the proportion of CD19(+)CD5(+) lymphocytes may serve as a cellular biomarker to predict the response to corticosteroid in LOHC.</p>


Subject(s)
Adolescent , Adrenal Cortex Hormones , Therapeutic Uses , Adult , Antigens, CD19 , Metabolism , B-Lymphocytes , Metabolism , CD5 Antigens , Metabolism , Child , Child, Preschool , Cystitis , Drug Therapy , Allergy and Immunology , Therapeutics , Female , Flow Cytometry , Hematopoietic Stem Cell Transplantation , Humans , Male , Middle Aged , Young Adult
11.
Article in English | WPRIM | ID: wpr-131144

ABSTRACT

BACKGROUND: Since the recent introduction of radioimmunotherapy (RIT) using antibodies against cluster of differentiation (CD) 45 for the treatment of lymphoma, the clinical significance of the CD45 antigen has been increasing steadily. Here, we analyzed CD45 expression on lymphocyte subsets using flow cytometry in order to predict the susceptibility of normal lymphocytes to RIT. METHODS: Peripheral blood specimens were collected from 14 healthy individuals aged 25-54 yr. The mean fluorescence intensity (MFI) of the cell surface antigens was measured using a FACSCanto II system (Becton Dickinson Bioscience, USA). MFI values were converted into antibody binding capacity values using a Quantum Simply Cellular microbead kit (Bangs Laboratories, Inc., USA). RESULTS: Among the lymphocyte subsets, the expression of CD45 was the highest (725,368+/-42,763) on natural killer T (NKT) cells, 674,030+/-48,187 on cytotoxic/suppressor T cells, 588,750+/-48,090 on natural killer (NK) cells, 580,211+/-29,168 on helper T (Th) cells, and 499,436+/-21,737 on B cells. The Th cells and NK cells expressed a similar level of CD45 (P=0.502). Forward scatter was the highest in NKT cells (P<0.05), whereas side scatter differed significantly between each of the lymphocyte subsets (P<0.05). CD3 expression was highest in the Th and NKT cells. CONCLUSIONS: NKT cells express the highest levels of CD45 antigen. Therefore, this lymphocyte subset would be most profoundly affected by RIT or pretargeted RIT. The monitoring of this lymphocyte subset during and after RIT should prove helpful.


Subject(s)
Adult , Antibodies/immunology , Leukocyte Common Antigens/analysis , B-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Female , Flow Cytometry/methods , Fluorescein-5-isothiocyanate/chemistry , Fluorescent Dyes/chemistry , Humans , Killer Cells, Natural/immunology , Lymphocytes/immunology , Lymphoma/radiotherapy , Male , Middle Aged , Natural Killer T-Cells/immunology , Protein Binding , Radioimmunotherapy , Reagent Kits, Diagnostic , T-Lymphocytes, Helper-Inducer/immunology
12.
Article in English | WPRIM | ID: wpr-131141

ABSTRACT

BACKGROUND: Since the recent introduction of radioimmunotherapy (RIT) using antibodies against cluster of differentiation (CD) 45 for the treatment of lymphoma, the clinical significance of the CD45 antigen has been increasing steadily. Here, we analyzed CD45 expression on lymphocyte subsets using flow cytometry in order to predict the susceptibility of normal lymphocytes to RIT. METHODS: Peripheral blood specimens were collected from 14 healthy individuals aged 25-54 yr. The mean fluorescence intensity (MFI) of the cell surface antigens was measured using a FACSCanto II system (Becton Dickinson Bioscience, USA). MFI values were converted into antibody binding capacity values using a Quantum Simply Cellular microbead kit (Bangs Laboratories, Inc., USA). RESULTS: Among the lymphocyte subsets, the expression of CD45 was the highest (725,368+/-42,763) on natural killer T (NKT) cells, 674,030+/-48,187 on cytotoxic/suppressor T cells, 588,750+/-48,090 on natural killer (NK) cells, 580,211+/-29,168 on helper T (Th) cells, and 499,436+/-21,737 on B cells. The Th cells and NK cells expressed a similar level of CD45 (P=0.502). Forward scatter was the highest in NKT cells (P<0.05), whereas side scatter differed significantly between each of the lymphocyte subsets (P<0.05). CD3 expression was highest in the Th and NKT cells. CONCLUSIONS: NKT cells express the highest levels of CD45 antigen. Therefore, this lymphocyte subset would be most profoundly affected by RIT or pretargeted RIT. The monitoring of this lymphocyte subset during and after RIT should prove helpful.


Subject(s)
Adult , Antibodies/immunology , Leukocyte Common Antigens/analysis , B-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Female , Flow Cytometry/methods , Fluorescein-5-isothiocyanate/chemistry , Fluorescent Dyes/chemistry , Humans , Killer Cells, Natural/immunology , Lymphocytes/immunology , Lymphoma/radiotherapy , Male , Middle Aged , Natural Killer T-Cells/immunology , Protein Binding , Radioimmunotherapy , Reagent Kits, Diagnostic , T-Lymphocytes, Helper-Inducer/immunology
13.
Braz. j. med. biol. res ; 43(12): 1215-1224, Dec. 2010. ilus, tab
Article in English | LILACS | ID: lil-568996

ABSTRACT

Rubinstein-Taybi syndrome (RTS) is a rare developmental disorder characterized by craniofacial dysmorphisms, broad thumbs and toes, mental and growth deficiency, and recurrent respiratory infections. RTS has been associated with CREBBP gene mutations, but EP300 gene mutations have recently been reported in 6 individuals. In the present study, the humoral immune response in 16 RTS patients with recurrent respiratory infections of possible bacterial etiology was evaluated. No significant differences between patients and 16 healthy controls were detected to explain the high susceptibility to respiratory infections: normal or elevated serum immunoglobulin levels, normal salivary IgA levels, and a good antibody response to both polysaccharide and protein antigens were observed. However, most patients presented high serum IgM levels, a high number of total B cell and B subsets, and also high percentiles of apoptosis, suggesting that they could present B dysregulation. The CREBBP/p300 family gene is extremely important for B-cell regulation, and RTS may represent an interesting human model for studying the molecular mechanisms involved in B-cell development.


Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Male , Young Adult , Antibodies, Monoclonal/analysis , B-Lymphocytes/immunology , Immunity, Humoral/immunology , Immunoglobulins/analysis , Respiratory Tract Infections/immunology , Rubinstein-Taybi Syndrome/immunology , Antibodies, Monoclonal/immunology , Case-Control Studies , CREB-Binding Protein/genetics , Immunity, Humoral/genetics , Immunoglobulins/immunology , Recurrence
14.
Chinese Medical Journal ; (24): 942-948, 2010.
Article in English | WPRIM | ID: wpr-242541

ABSTRACT

<p><b>BACKGROUND</b>Rapamycin (RAPA) is a relatively new immunosuppressant drug that functions as a serine/threonine kinase inhibitor to prevent rejection in organ transplantation. RAPA blocks activation of T-effector (Teff) cells by inhibiting the response to interleukin-2. Recently, RAPA was also shown to selectively expand the T-regulator (Treg) cell population. To date, no studies have examined the mechanism by which RAPA converts Teff cells to Treg cells.</p><p><b>METHODS</b>Peripheral CD4(+)CD25(-) naive T cells were cultivated with RAPA and B cells as antigen-presenting cells (APCs) in vitro. CD4(+)CD25(-) T cells were harvested after 6 days and analyzed for expression of forkhead box protein 3 (Foxp3) using flow cytometry. CD4(+)CD25(+)CD127(-) subsets as the converted Tregs were isolated from the mixed lymphocyte reactions (MLR) with CD127 negative selection, followed by CD4 and CD25 positive selection using microbeads and magnetic separation column (MSC). Moreover, mRNA was extracted from converted Tregs and C57BL/6 naive CD4(+)CD25(+) T cells and Foxp3 levels were examined by quantitative real-time polymerase chain reaction (rt-PCR). A total of 1 x 10(5) carboxyfluorescein succinimidyl ester (CFSE)-labeled naive CD4(+)CD25(-) T cells/well from C57BL/6 mice were cocultured with DBA/2 or C3H maturation of dendritic cells (mDCs) (0.25 x 10(5)/well) in 96-well round-bottom plates for 6 days. Then 1 x 10(5) or 0.25 x 10(5) converted Treg cells were added to every well as regulatory cells. Cells were harvested after 6 days of culture and analyzed for proliferation of CFSE-labeled naive CD4(+)CD25(-) T cells using flow cytometry. Data were analyzed using CellQuest software.</p><p><b>RESULTS</b>We found that RAPA can convert peripheral CD4(+)CD25(-) naive T Cells to CD4(+)Foxp3(+) Treg cells using B cells as APCs, and this subtype of Treg can potently suppress Teff proliferation and maintain antigenic specificity.</p><p><b>CONCLUSION</b>Our findings provide evidence that RAPA induces Treg cell conversion from Teff cells and uncovers an additional mechanism for tolerance induction by RAPA.</p>


Subject(s)
Animals , Antibiotics, Antineoplastic , Pharmacology , Antigen-Presenting Cells , Allergy and Immunology , Metabolism , B-Lymphocytes , Allergy and Immunology , Metabolism , CD4-Positive T-Lymphocytes , Allergy and Immunology , Metabolism , Cell Proliferation , Dendritic Cells , Allergy and Immunology , Metabolism , Forkhead Transcription Factors , Metabolism , Interleukin-2 Receptor alpha Subunit , Metabolism , Interleukin-7 Receptor alpha Subunit , Metabolism , Male , Mice , Mice, Inbred C57BL , Mice, Inbred DBA , Mitomycin , Pharmacology , Polymerase Chain Reaction , Sirolimus , Pharmacology , T-Lymphocytes, Regulatory , Allergy and Immunology , Metabolism
15.
Journal of Experimental Hematology ; (6): 1079-1083, 2010.
Article in Chinese | WPRIM | ID: wpr-237591

ABSTRACT

Mesenchymal stem cells (MSCs) can inhibit T cell proliferation, the effects of MSCs on various T cell subsets have showed different immune regulatory reactions, and their mechanisms mainly include cell-cell contact and mediation by cytokines secreted from MSCs. Encouragingly, recent studies have showed that the effects of MSCs on T-cell response to pathogens is not significant, but can obviously suppress T cell response to allogeneic antigens. In addition, MSCs can regulate the proliferation, survival, antibody secretion and differentiation of B cells, inhibit the production, proliferation, migration and antigen-presentation of DCs, and modulate the differentiation and maturation of DCs, and regulate the proliferation, cell toxicity and cytokine secretion of NK cells. In this review, the research advances on immunomodulatory effects of MSCs on various immune cells including T-lymphocytes, B-lymphocytes, NK cells and DCs are summarized with emphasis on the immunoregulatory effects of MSCs on T-lymphocytes.


Subject(s)
B-Lymphocytes , Allergy and Immunology , Dendritic Cells , Allergy and Immunology , Humans , Killer Cells, Natural , Allergy and Immunology , Mesenchymal Stem Cells , Cell Biology , Allergy and Immunology , T-Lymphocytes , Allergy and Immunology
16.
Chinese Medical Journal ; (24): 1784-1789, 2009.
Article in English | WPRIM | ID: wpr-240797

ABSTRACT

<p><b>BACKGROUND</b>The role of B-cell remains an enigma in the pathogenesis of ankylosing spondylitis (AS). This study aimed to investigate the distributions of B-cells and subsets in peripheral blood of AS patients and observe their changes in etanercept-treated AS patents.</p><p><b>METHODS</b>We detected the proportions of CD19(+) B-cell, naive B-cell (CD19(+)CD27-), memory B-cell (CD19(+)CD27dim) and plasmablast (CD19(+)CD27high) in peripheral blood of 66 patients with AS (39 at active stage, 27 at stable stage; 35 patients with peripheral joint involvement, 31 patients with axial involvement alone), 30 patients with rheumatoid arthritis (RA) and 30 healthy volunteers using flow cytometry. And then we observed the changes of the above indexes of 39 active AS patients treated with etanercept in a randomized, double-blind, placebo-controlled trial.</p><p><b>RESULTS</b>(1) Percentages of CD19(+) B-cells in active or peripheral joint involvement AS patients increased more obviously than those in stable or axial involvement alone AS patients (both P = 0.001), and percentage of CD19(+)CD27high B-cells in AS patients with peripheral joint involvement was significantly higher than that in cases with axial involvement alone or healthy volunteers (P = 0.005 and 0.006, respectively); (2) The percentage of CD19(+) B-cells in AS patients was positively correlated with Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) scores, Patient's Global Assessment (PGA) scores, total back pain scores and nocturnal back pain scores (r = 0.270, 0.255, 0.251 and 0.266, P = 0.029, 0.039, 0.042 and 0.031, respectively); (3) At week 6 and week 12, there were no statistical differences of the percentages of B-cells and subsets between etanercept group and placebo group of AS patients (P > 0.05); the percentage of CD19(+) B-cells in etanercept group was higher than that in healthy volunteers at week 12 (t = 3.320, P = 0.003).</p><p><b>CONCLUSIONS</b>Misbalance is present in B-cells and some subsets in peripheral blood of active AS patients with peripheral joint involved. B-cells might play an important role in the pathogenesis of AS patients. The high percentage of CD19(+) B-cells in active AS patients cannot be down-regulated after 12-week etanercept treatment.</p>


Subject(s)
Adolescent , Adult , Antigens, CD19 , Allergy and Immunology , B-Lymphocytes , Allergy and Immunology , Etanercept , Female , Flow Cytometry , Humans , Immunoglobulin G , Pharmacology , Therapeutic Uses , Immunosuppressive Agents , Pharmacology , Therapeutic Uses , Male , Middle Aged , Receptors, Tumor Necrosis Factor , Therapeutic Uses , Spondylitis, Ankylosing , Drug Therapy , Allergy and Immunology , Tumor Necrosis Factor Receptor Superfamily, Member 7 , Allergy and Immunology , Young Adult
17.
São Paulo; s.n; 16 dez. 2008. 137[22] p. ilus, graf, tab.
Thesis in Portuguese | LILACS | ID: lil-512972

ABSTRACT

A cavidade peritoneal de camundongos abriga uma variedade de células do sistema imune. Inicialmente, devido as limitações metodológicas, acreditava-se que, aproximadamente, 90% das células peritoneais era representada por macrófagos. Em seguida, graças aos extensos estudos com células peritoneais, observou-se que, além dos macrófagos, o peritônio abrigava muitos Linfócitos B, principalmente do subtipo B-1. Utilizando metodologias contemporâneas de FACS - citometria de fluxo, este trabalho mostra que, aproximadamente, 30% das células peritoneais são macrófagos, 55% são Iinfócitos B-1, dos quais 40% pertencem ao subtipo B-1a e 15% ao subtipo B-1b. Os 15% - 20% restantes representam outros subtipos celulares, como linfócitos T, linfócitos B-2, linfócitos NK, eosinófilos, além da presença de outras populações de células que não foram possíveis de ser identificadas com os marcadores de superfície utilizados neste trabalho. Em contraste com a literatura, nossos estudos mostraram que os macrófagos da cavidade peritoneal de camundongos representam uma população heterogênea...


Subject(s)
Mice , Peritoneal Cavity/cytology , In Vitro Techniques , B-Lymphocytes/cytology , B-Lymphocytes/immunology , Macrophages, Peritoneal/cytology , Macrophages, Peritoneal/immunology , Phenotype , Biological Assay/methods , Flow Cytometry , Microscopy, Confocal/methods , Microscopy, Electron/methods
18.
Medicina (B.Aires) ; 68(6): 423-427, nov.-dic. 2008. tab
Article in English | LILACS | ID: lil-633581

ABSTRACT

The purpose of this study was to characterize and quantify cells involved in immune response in metastasis-free regional lymph nodes (RLNs) draining different human epithelial tumors and compare them (by immunohistochemistry) with control lymph nodes from patients with non malignant diseases. We showed that T cells number was decreased in RLNs as compared to the controls with reduction in both CD4+ T cells and CD8+ T cells subsets and an inverted ratio (CD4+: CD8+). B lymphocytes and follicular dendritic cells were decreased with respect to the controls. S100+ dendritic cells (DCs) and mature DCs were detected in T dependent areas. Their mean number was significantly lower as compared to control. Immature DCs were significantly diminished compared to RLN and control nodes. CD57+ cells, follicular T helper cells and/or NK cells, were localized in the clear zone of germinal centres and their mean number was significantly increased. There were no CD57+ cells in hypoplastic follicles. In this study we show that RLNs draining human cancer present reduction in almost all immune cells, except CD57+ cells. These findings may be related to the deficient anti-tumor immune response in patients with cancer and subsequent tumor progression.


El objetivo del trabajo fue caracterizar y cuantificar utilizando inmuno-histoquímica, las células involucradas en la respuesta inmune en ganglios linfáticos regionales (GLRs) que drenan distintos tumores epiteliales malignos humanos y compararlas con ganglios controles (GLCs) provenientes de pacientes sin enfermedad neoplásica maligna. Determinamos que los GLRs presentaban una marcada depleción de linfocitos B y T, células dendríticas (CD) foliculares y CD interdigitantes maduras respecto a los controles. En los linfocitos T, además de estar disminuidos, se observó una inversión de la relación T CD4+: T CD8+, a favor de los T CD8+. La depleción de CD inmaduras fue mayor respecto a las maduras. Las células CD57+, células foliculares T helper y/o células NK, localizadas en las zonas claras de los centros germinativos, presentaron un marcado incremento en los GLRs comparados con los GLCs, excepto en los casos de ganglios linfáticos con folículos hipoplásicos. En este estudio, demostramos que los GLRs que drenan carcinomas humanos presentan una significativa reducción en casi todas las células de la respuesta inmune, excepto las células NK. Estos hallazgos podrían estar relacionados con la deficiente respuesta antitumoral de los pacientes con cáncer y la subsiguiente progresión tumoral.


Subject(s)
Adult , Aged , Humans , Middle Aged , Dendritic Cells/cytology , Dendritic Cells/immunology , Lymph Nodes/immunology , Lymphocyte Subsets/immunology , Neoplasms/immunology , T-Lymphocytes/cytology , B-Lymphocytes/immunology , Case-Control Studies , Immunity, Cellular , Lymph Nodes/pathology , Lymphatic Metastasis/immunology , Lymphatic Metastasis/pathology , Neoplasms/pathology , T-Lymphocytes/immunology
19.
IPMJ-Iraqi Postgraduate Medical Journal. 2008; 7 (2): 112-120
in English | IMEMR | ID: emr-108449

ABSTRACT

In Type 1 Diabetes Mellitus [T1DM], numerous changes in the cellular as well humoral immune response have been identified. However, it is not known whether both the CD[4+] and CD[8+] subpopulation or only one of these or CD[19+]contains increased numbers of activated cells. The aim was to study the activated lymphocyte subpopulation by use of monoclonal antibodies to T-cell and B-cell antigens which is known to be expressed on activated cells. A total of 60 T1DM patients who had newly onset of the disease [diagnosed was from one week up to five months] were included in the present study, all the patients were treated with daily replacement doses of insulin. Fifty apparently healthy control subjects underwent the PBL phenotyping. Phenotyping of surface antigens was done by direct Immunoflurocent [IFT] technique using mouse antihuman CD[3], CD[4], CD[8], CD[45]RA, CD[19], and activated markers CD[45]RO, DR-antigen and CD[38]. T1DM patients showed a remarkable lowering in CD[3+], CD[8+], and CD[45]RA[+] cells [p<0.0001], but the decrease in CD[4+] cells percentage was not significant. In contrast, a significant elevation of activation markers includes [CD[45]RO[+], HLA-DR[+] and CD[38+] cells] were observed in patients in addition to a significant increase of CD[19+] cell percentage and CD[4+]: CD[8+] ratio in the patients. This study provides evidence that abnormalities of T-cells regulation are detectable in patients with T1DM


Subject(s)
Humans , Male , Female , Child, Preschool , Adolescent , Child , Diabetes Mellitus, Type 1/immunology , Lymphocyte Subsets , CD4-Positive T-Lymphocytes , CD8-Positive T-Lymphocytes , B-Lymphocytes
20.
Article in Chinese | WPRIM | ID: wpr-270682

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of rapamycin in inducing naïve murine effector T cell (Teff) convert to regulatory T cell (Treg) in vitro.</p><p><b>METHODS</b>The forkhead box protein 3 (FoxP3) negative Teff were isolated and purified from the spleen and lymph node of C57 BL/6 murines aged 6-8 weeks, then Teff were cultured in three groups with mature dendritic cells (mDC), B cells, and plate coated Anti-CD3. In addition, the control wells and the test wells were prepared in each group, rapamycin were not added in the control wells but added in the test wells with concentrations of 1, 10, 50, and 100 nmol/L. Percentages of FoxP3 positive Treg were examined by flow cytometry after 4 days in Anti-CD3 group and after 6 days in the other two groups.</p><p><b>RESULTS</b>As shown by the flow cytometry, the percentages of FoxP3 positive Treg were as follows in three group: in the mDC group, it was 0.01% in the control well and 0.39%, 0.47%, 0.34%, and 0.26% in test wells; in B cell group, it was 0.01% in the control wells and 5.56%, 5.89%, 7.15%, and 4.72% in the test wells; in Anti-CD3 group, it was 0.93% in the control wells and 1.35%, 1.07%, 1.02%, and 1.19% in test wells. No significant difference was found between the test wells and control wells in the mDC group and Anti-CD3 group; however, the percentages of FoxP3 positive Treg was significantly different between the test wells and control wells in the B cell group (P < 0.01).</p><p><b>CONCLUSION</b>When B cell is acted as the antigen-presenting cell, rapamycin can effectively induce Teff convert to Treg in vitro.</p>


Subject(s)
Animals , B-Lymphocytes , Cell Biology , Allergy and Immunology , Cell Differentiation , Cells, Cultured , Dendritic Cells , Cell Biology , Allergy and Immunology , Flow Cytometry , Forkhead Transcription Factors , Allergy and Immunology , Mice , Mice, Inbred C57BL , Mice, Inbred DBA , Precursor Cells, T-Lymphoid , Cell Biology , Allergy and Immunology , Sirolimus , Pharmacology , T-Lymphocyte Subsets , Cell Biology , Allergy and Immunology , T-Lymphocytes, Regulatory , Cell Biology , Allergy and Immunology
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