1.Immunologic function of palatine tonsil B lymphocyte.
Mike MIN ; Chaowu MA ; Boquan JIN ; Jianzhong XU ; Yu ZHOU ; Xinfei GAN
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2009;23(7):311-315
OBJECTIVE:
To study cellular immune function of palatine tonsil B lymph cell.
METHOD:
The phenotype of palatine tonsil cells (PTC) and that of peripheral blood mononuclear cell (PBMC) were compared using fluorescence staining and flow cytometry (FCM) analysis, then immunomagnetic beads were used to separate CD3- cell in PTC and PBMC. The proliferation function of CD3- lymph cell of PTC and PBMC was tested after stimulated by CD20mAb.
RESULT:
FCM analysis founding that 71.2% PTC express CD20 with higher mean fluorescence intensity, MFI, compared to the 15.5% in PBMC. There's no significant difference between the proliferation of PTC and PBMC B lymph cell.
CONCLUSION
CD20 expression is different in PTC and PBMC, but corresponding function is still unknown.
Adult
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Antigens, CD20
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metabolism
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B-Lymphocytes
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cytology
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immunology
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metabolism
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Flow Cytometry
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Humans
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Palatine Tonsil
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cytology
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immunology
2.Inhibitory function of Tregs via soluble FGL2 in chronic hepatitis B.
Li XU ; Daofeng YANG ; Yanlin LIU ; Di WU ; Xiaojing WANG ; Qin NING
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(4):540-545
CD4(+)CD25(+)CD127(dim/-) regulatory T cells (Tregs) have been implicated in suppressing T cell immune responses to hepatitis B virus (HBV), but the inhibition mechanism has not being clear yet. This study investigated the effects of soluble FGL2 (sFGL2) secreted by Tregs on immune suppression in chronic HBV-infected patients. We verified that sFGL2 protein and mRNA were highly expressed in Tregs. The separated Tregs by using magnetic beads from peripheral blood mononuclear cells (PBMCs) in 20 patients with chronic hepatitis B were co-cultured with PBMCs at a ratio of 1:3 with anti-CD3 stimulating antibody or FGL2 blocking antibody. The proliferation index of CD8(+)T cells after blocking FGL2 was higher than that in blank group (3.58±0.18 vs. 3.28±0.17, P=0.034) in 18 of 20 samples, and lower than that in CD3 stimulation group (3.82±0.19, P=0.026) in 16 of 20 samples. The IFN-γ secreted in the mixed culture in the absence of Tregs was higher than that in the culture in the presence of Tregs, but it could be abolished by FGL2 blocking antibody. These results suggest that sFGL2 protein secreted by Tregs suppresses the proliferation and function of CD8(+) T cells in chronic hepatitis B.
CD8-Positive T-Lymphocytes
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immunology
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metabolism
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Cells, Cultured
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Fibrinogen
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immunology
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metabolism
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Hepatitis B, Chronic
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immunology
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metabolism
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Humans
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T-Lymphocytes, Regulatory
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immunology
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metabolism
3.Single B cell monoclonal antibody technologies and applications.
Xiangyang CHI ; Changming YU ; Wei CHEN
Chinese Journal of Biotechnology 2012;28(6):651-660
Monoclonal antibodies (mAbs) contribute a lot to the development of numerous fields in life science as a pivotal tool in modern biological research. Development of the PCR methods and maturation of antibody production have made it possible to generate mAbs from single human B cells by single cell RT-PCR with successional cloning and expression in vitro. Compared to traditional monoclonal antibody technologies, single B cell technologies require relatively fewer cells, which are highly efficient in obtaining specific mAbs in a rapid way with preservation of the natural heavy and light chain pairing. With so many advantages, single B cell technologies have been proved to be an attractive approach for retrieval of naive and antigen-experienced antibody repertoires generated in vivo, design of rationale structure-based vaccine, evaluation and development of basic B cell biology concepts in health and autoimmunity, and prevention of infectious diseases by passive immunization and therapy for disorders. Accordingly, this review introduced recent progresses in the single B cell technologies for generating monoclonal antibodies and applications.
Antibodies, Monoclonal
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biosynthesis
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genetics
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immunology
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Antibody Specificity
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B-Lymphocytes
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cytology
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immunology
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metabolism
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Humans
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Immunologic Techniques
4.Abnormal expression of CD43 in patients with systemic lupus erythematosus and its clinical significance.
Li-xia XIA ; Xin DING ; Xin LI ; Feng-chun ZHANG ; Fu-lin TANG ; Xuan ZHANG ; De-nian BA ; Wei HE
Chinese Medical Journal 2012;125(7):1266-1271
<b>BACKGROUNDb>Previous studies indicate that CD43 plays a role in regulating the adhesion of lymphocytes, cell mutation and activation, however, little is known about its effect on systemic lupus erythematosus (SLE). This study was designed to explore the clinical significance of CD43 in SLE patients.
<b>METHODSb>We used microarray and real-time PCR to detect the mRNA and protein expression of magnetic bead sorted T cells and B cells from peripheral blood mononuclear cells (PBMCs) of SLE patients, and analyzed the relationship between CD43 and the clinical indexes.
<b>RESULTSb>Both microarray and real-time PCR results showed that CD43 mRNA was significantly decreased in PBMCs of SLE patients compared with healthy controls (P < 0.001). There were no significant differences between lupus nephritis and non-lupus nephritis patients, and neuropsychiatric and non-neuropsychiatric patients. CD43 mRNA expression was significantly reduced in T cells but not in B-cells in SLE patients compared to healthy controls (P < 0.01). Compared with healthy controls, the percentage of CD43(+) cells in the PBMCs of SLE was significantly decreased (P = 0.004), and the CD43 fluorescence intensity in CD3(+)/CD43(+) cells and CD19(+)/CD43(+) cells was also significantly weaker than in healthy controls (P = 0.039 and 0.003). There was no significant difference in the percentage of CD3(+)/CD43(+) cells, CD19(+)/CD43(+) cells between the two groups. The CD43 fluorescence intensity in CD3(+)/CD43(+) cells was inversely correlated with the levels of IgG and IgM (r = -0.8 and -0.6).
<b>CONCLUSIONSb>Compared to healthy controls, both CD43 mRNA and protein expressions were reduced in T cells from patients with SLE, and were inversely correlated with IgG.
B-Lymphocytes ; immunology ; metabolism ; Humans ; Leukocytes, Mononuclear ; Leukosialin ; genetics ; metabolism ; Lupus Erythematosus, Systemic ; immunology ; metabolism ; Oligonucleotide Array Sequence Analysis ; Real-Time Polymerase Chain Reaction ; T-Lymphocytes ; immunology ; metabolism
6.Effects of Laughter Therapy on Depression, Quality of Life, Resilience and Immune Responses in Breast Cancer Survivors.
Journal of Korean Academy of Nursing 2011;41(3):285-293
PURPOSE: In this study, the effects of laughter therapy on levels of depression, quality of life, resilience and immune responses in breast cancer survivors were examined. METHODS: A quasi-experimental nonequivalent control group, pretest-posttest design was used. Participants (n=37) included breast cancer survivors who finished chemotheraphy and radiation therapy: 16 in the experiment group and 21 in the control group. Data were collected from August to November 2009. The experimental group participated in laughter therapy eight times, twice a week for 60 min per session. Questionnaires were used to me-asure pretest and posttest levels of depression, quality of life and resilience. A blood test was used to analyze changes in Total T cell, T helper, T suppressor, Th/Ts ratio, Total B cell, T cell/B cell ratio and NK cell for immune responses. RESULTS: The results showed that laughter therapy was effective in increasing the quality of life and resilience in breast cancer survivors. but depression and immune responses did not differ significantly between the groups. CONCLUSION: The results of the study indicate that laughter therapy may be an effective nursing intervention to improve quality of life and resilience in breast cancer survivors.
Adult
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Aged
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B-Lymphocytes/immunology/metabolism
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Breast Neoplasms/immunology/*therapy
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*Depression
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Female
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Humans
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*Laughter Therapy
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Middle Aged
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*Quality of Life
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Questionnaires
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*Resilience, Psychological
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T-Lymphocytes/immunology/metabolism
7.Comparative Quantitative Analysis of Cluster of Differentiation 45 Antigen Expression on Lymphocyte Subsets.
Mijeong IM ; Hyojin CHAE ; Taehoon KIM ; Hun Hee PARK ; Jihyang LIM ; Eun Jee OH ; Yonggoo KIM ; Yeon Joon PARK ; Kyungja HAN
The Korean Journal of Laboratory Medicine 2011;31(3):148-153
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.
Adult
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Antibodies/immunology
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Antigens, CD45/*analysis/immunology
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B-Lymphocytes/immunology/metabolism
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CD8-Positive T-Lymphocytes/immunology/metabolism
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Female
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Flow Cytometry/*methods
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Fluorescein-5-isothiocyanate/chemistry
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Fluorescent Dyes/chemistry
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Humans
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Killer Cells, Natural/immunology/metabolism
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Lymphocytes/immunology/*metabolism
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Lymphoma/radiotherapy
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Male
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Middle Aged
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Natural Killer T-Cells/immunology/metabolism
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Protein Binding
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Radioimmunotherapy
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Reagent Kits, Diagnostic
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T-Lymphocytes, Helper-Inducer/immunology/metabolism
8.Comparative Quantitative Analysis of Cluster of Differentiation 45 Antigen Expression on Lymphocyte Subsets.
Mijeong IM ; Hyojin CHAE ; Taehoon KIM ; Hun Hee PARK ; Jihyang LIM ; Eun Jee OH ; Yonggoo KIM ; Yeon Joon PARK ; Kyungja HAN
The Korean Journal of Laboratory Medicine 2011;31(3):148-153
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.
Adult
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Antibodies/immunology
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Antigens, CD45/*analysis/immunology
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B-Lymphocytes/immunology/metabolism
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CD8-Positive T-Lymphocytes/immunology/metabolism
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Female
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Flow Cytometry/*methods
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Fluorescein-5-isothiocyanate/chemistry
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Fluorescent Dyes/chemistry
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Humans
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Killer Cells, Natural/immunology/metabolism
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Lymphocytes/immunology/*metabolism
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Lymphoma/radiotherapy
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Male
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Middle Aged
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Natural Killer T-Cells/immunology/metabolism
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Protein Binding
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Radioimmunotherapy
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Reagent Kits, Diagnostic
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T-Lymphocytes, Helper-Inducer/immunology/metabolism
9.Expression of B7-H1 on peripheral blood mononuclear cells from chronic hepatitis B patients.
Zhun-yi XIE ; Yong-wen CHEN ; Xiao-lan FU ; Cheng-ying YANG ; Yu-zhang WU
Journal of Southern Medical University 2007;27(11):1635-1637
<b>OBJECTIVEb>To investigate the expression of B7-H1 on peripheral blood mononuclear cells from patients with chronic HBV infection.
<b>METHODSb>Immunofluorescent staining and flow cytometry assay were used to measure the expression of B7-H1 on peripheral blood CD3high T cells, CD19high B cells and CD14high monocytes from chronic HBV infected patients.
<b>RESULTSb>No significant difference was observed in B7-H1 expression on T cells and B cells between chronic HBV infected patients (CHB) and health controls (HC). B7-H1-expressing CD14high cells were significantly increased in chronic HBV-infected patients (19.17-/+11.64)% as compared with healthy controls [(7.30-/+5.49)%, P<0.01]. A significant positive correlation was found between B7-H1 expression on CD14high monocytes and serum ALT levels.
<b>CONCLUSIONb>There is no significant difference in B7-H1 expression on T cells and B cells between CHB patients and healthy subjects. B7-H1, which is up-regulated on monocytes from chronic HBV-infected patients, in positively correlated to serum ALT levels, and may play a role in the persistence of HBV infection.
Adult ; Aged ; Antigens, CD ; metabolism ; B-Lymphocytes ; immunology ; B7-H1 Antigen ; Case-Control Studies ; Female ; Hepatitis B, Chronic ; immunology ; metabolism ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; Monocytes ; immunology ; T-Lymphocytes ; immunology ; Young Adult
10.Perspectives on the role of mTORC2 in B lymphocyte development, immunity and tumorigenesis.
Protein & Cell 2011;2(7):523-530
Mammalian target of rapamycin complex 2 (mTORC2) is a key downstream mediator of phosphoinositol-3-kinase (PI3K) dependent growth factor signaling. In lymphocytes, mTORC2 has emerged as an important regulator of cell development, homeostasis and immune responses. However, our current understanding of mTORC2 functions and the molecular mechanisms regulating mTORC2 signaling in B and T cells are still largely incomplete. Recent studies have begun to shed light on this important pathway. We have previously reported that mTORC2 mediates growth factor dependent phosphorylation of Akt and facilitates Akt dependent phosphorylation and inactivation of transcription factors FoxO1 and FoxO3a. We have recently explored the functions of mTORC2 in B cells and show that mTORC2 plays a key role in regulating survival and immunoglobulin (Ig) gene recombination of bone marrow B cells through an Akt2-FoxO1 dependent mechanism. Ig recombination is suppressed in proliferating B cells to ensure that DNA double strand breaks are not generated in actively dividing cells. Our results raise the possibility that genetic or pharmacologic inhibition of mTORC2 may promote B cell tumor development as a result of inefficient suppression of Ig recombination in dividing B cells. We also propose a novel strategy to treat cancers based on our recent discovery that mTORC2 regulates Akt protein stability.
B-Lymphocytes
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cytology
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immunology
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Cell Differentiation
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Cell Transformation, Neoplastic
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Humans
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Transcription Factors
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metabolism