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 ; Antigens, CD20 ; metabolism ; B-Lymphocytes ; cytology ; immunology ; metabolism ; Flow Cytometry ; Humans ; Palatine Tonsil ; cytology ; immunology

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 ; biosynthesis ; genetics ; immunology ; Antibody Specificity ; B-Lymphocytes ; cytology ; immunology ; metabolism ; Humans ; Immunologic Techniques

Flow cytometry was used to identify and characterize monoclonal antibodies (mAbs) that react with rabbit leukocyte differentiation molecules (LDM). Screening sets of mAbs, developed against LDM in other species, for reactivity with rabbit LDM yielded 11 mAbs that recognize conserved epitopes on rabbit LDM orthologues and multiple mAbs that recognize epitopes expressed on the major histocompatibility class I or class II molecules. Screening of mAbs submitted to the Animal Homologues Section of the Eighth Human Leukocyte Differentiation Workshop yielded 7 additional mAbs. Screening of mAbs generated from mice immunized with leukocytes from rabbit thymus or spleen or concanavalin A activated peripheral blood and/or spleen lymphocytes has yielded 42 mAbs that recognize species restricted epitopes expressed on one or more lineages of leukocytes. Screening of the anti-rabbit mAbs against leukocytes from other species yielded one additional mAb. The studies show that screening of existing sets of mAbs for reactivity with rabbit LDM will not be productive and that a direct approach will be needed to develop mAbs for research in rabbits. The flow cytometric approach we developed to screen for mAbs of interest offers a way for individual laboratories to identify and characterize mAbs to LDM in rabbits and other species. A web-based program we developed provides a source of information that will facilitate analysis. It contains a searchable data base on known CD molecules and a data base on mAbs, known to react with LDM in one or more species of artiodactyla, equidae, carnivora, and or lagomorpha.
Animals ; Antibodies, Monoclonal/*immunology ; Antigens, Differentiation/*metabolism ; B-Lymphocytes/cytology/metabolism ; Basophils/cytology/metabolism ; Epitopes/genetics/metabolism ; *Flow Cytometry ; Gene Expression Regulation ; Granulocytes/cytology/metabolism ; Leukocytes/immunology/*metabolism ; Mice ; Monocytes/cytology/metabolism ; Rabbits ; T-Lymphocytes/cytology/metabolism

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 ; cytology ; immunology ; Cell Differentiation ; Cell Transformation, Neoplastic ; Humans ; Transcription Factors ; metabolism

<b>OBJECTIVEb>To assess the prognostic value of CD20(+) tumor-infiltrating lymphocytes (TILs) in early-stage breast cancer.

<b>METHODSb>Paraffin sections were collected from 130 cases of stage I-III breast cancer undergoing surgery between January, 2000 and December, 2002 in our hospital. Immunohistochemistry was used to analyze mesenchymal CD20(+) TILs infiltration in the tumor and evaluate its association with the density of CD4(+) and CD8(+) TILs. The association of CD20(+) TILs was evaluated with the histopathologic features, overall survival (OS), distant disease-free survival (DDFS), and disease-free survival (DFS) of the patients.

<b>RESULTSb>Aggregations of CD20(+) lymphocytes were observed in 37.69% (49/130) of the cases. CD3(+) T cells were found to aggregate around CD20(+) B cell aggregations to form lymphoid follicle-like structures. The aggregations of CD20(+) TILs were positively correlated with the densities of mesenchymal CD8+ and CD4(+) TILs. Overall, CD20(+) TIL aggregations were not significantly correlated with the outcomes of the patients, but multivariate COX regressions suggested that CD20(+) TIL aggregations were positively correlated with DDFS (HR=0.251, 95% CI=0.071-0.894, P=0.033) and OS (HR=0.325, 95% CI=0.103-1.028, P=0.056) in hormone receptor-negative patients but not in the positive patients. Further analysis suggested that post-operative adjuvant endocrine therapy significantly improved the OS of patients positive for hormone receptors without CD20(+) TIL aggregations (P=0.001).

<b>CONCLUSIONb>The long-term therapeutic effects of adjuvant endocrine therapy are correlated with CD20(+) TIL aggregations to affect prognostic value of CD20(+) TIL aggregations in early-stage breast cancer patients.

Antigens, CD20 ; metabolism ; B-Lymphocytes ; cytology ; Breast Neoplasms ; immunology ; pathology ; Disease-Free Survival ; Female ; Humans ; Lymphocytes, Tumor-Infiltrating ; cytology ; Middle Aged ; Neoplasm Invasiveness ; Prognosis ; Survival Rate

<b>AIMb>To investigate effects of hsBAFF synthesized in Escherichia coli on spleen B lymphocyte immune response and its intracellular free Ca2+ ([Ca2]i]) signaling in mice.

<b>METHODSb>Twenty ICR mice, half males-half females, were chosen and randomly divided into a normal control group (n=10) and a hsBAFF treatment group (n-10). The mice in hsBAFF treatment group were given abdominal cavity injection of hsBAFF solution which was diluted with phosphate buffered saline (PBS) at dosage of 0.1 mg/kg body weight once each day for over eight days. The mice in control group were received abdominal injection of PBS at the same dose and frequency. Spleen B lymphocyte proliferation and its immune response to LPS stimulation in mice were evaluated using an MTT assay, and change of spleen B lymphocyte [Ca2+]i was assayed under a laser scanning confocal microscope.

<b>RESULTSb>B lymphocyte proliferation and its immune response to LPS stimulation were significantly higher in hsBAFF-treated mice than in control mice (P < 0.05). The B lymphocyte [Ca2+]i fluorescence intensity in hsBAFF-treated mice maintained at a relatively high level fluctuation, and its average intensity was significantly higher to that of control mice (P < 0.01), but change rate of the intensity was lower compared to that of control group.

<b>CONCLUSIONb>hsBAFF synthesized in Escherichia coli can enhance immune function in the body by increasing B lymphocyte proliferation and its immune response. hsBAFF-activated B lymphocyte function may be associated with increasing B lymphocytes [Ca2+]i.

Animals ; B-Cell Activating Factor ; immunology ; pharmacology ; B-Lymphocytes ; cytology ; drug effects ; immunology ; Calcium ; metabolism ; Calcium Signaling ; drug effects ; Cell Proliferation ; drug effects ; Female ; Male ; Mice ; Mice, Inbred ICR ; Spleen ; cytology ; drug effects ; immunology

Adult ; Case-Control Studies ; Female ; Hepatitis B, Chronic ; immunology ; metabolism ; Humans ; Male ; Middle Aged ; T-Lymphocyte Subsets ; cytology ; T-Lymphocytes, Cytotoxic ; cytology

The OPG/RANKL/RANK system plays an important role in osteoclastogenesis and represents a great progress in bone biology. RANKL, which expresses on the surface of osteoblast/stromal cells and activated T cells, binds to RANK on the osteoclastic precursors or mature osteoclasts, and promotes osteoclastogenesis and bone resorption. While osteoprotegerin (OPG), which is expressed by osteoblasts/stromal cells, strongly inhibits bone resorption by binding to its ligand RANKL and thereby blocks the interaction between BANKL and RANK. A number of cytokines and hormones exert their effects on bone metabolism by regulating the OPG/RANKL ratio in the bone marrow microenvironment. RANK is also expressed on mammary epithelial cells and RANKL expression in these cells is induced by pregnancy hormones, RANKL and RANK are essential for the formation of the lactating mammary gland and the transmission of maternal calcium to neonates in mammalian species. Modulation of these systems provides a unique opportunity to develop novel therapeutics to inhibit bone loss in osteoporosis, rheumatoid arthritis, and bone metastasis of cancer. Further research should be focused on the cooperation of OPG/RANKL/RANK system with other signal pathways and the interactions among bone remodeling, immune system and endocrinology system. Currently, the development of OPG analogues or compounds which may stimulate OPG expression is becoming an attractive industry which may be profitable to both patients and manufacturers.
Animals ; Bone Resorption ; immunology ; metabolism ; Humans ; Osteoclasts ; cytology ; metabolism ; pathology ; Osteogenesis ; drug effects ; genetics ; immunology ; Osteoprotegerin ; metabolism ; physiology ; RANK Ligand ; metabolism ; physiology ; Receptor Activator of Nuclear Factor-kappa B ; metabolism ; pharmacology ; physiology ; T-Lymphocytes ; drug effects ; immunology

<b>OBJECTIVEb>To study activation of dendritic cells (DC) isolated from peripheral blood monocytes of patients with chronic hepatitis B after stimulation with HBsAg or HBcAg.

<b>METHODSb>DCs were isolated from peripheral blood monocytes of patients with chronic hepatitis B. DCs were impulsed with HBsAg and HBcAg separately before their maturation. The expression levels of DC surface molecule were analyzed by using flow cytometry and the ability of DC to induce T lymphocyte proliferation was evaluated by a liquid scintillation counter and the amount of interleukin-12 (IL-12) in mixed lymphocytic (IL-12) in mixed lymphocytic reaction (MLR) was measured by using ELISA.

<b>RESULTSb>The expression rate of CD86 significantly increased to (29.20 +/- 5.18)% on DC after loading with HBcAg as compared with those after loading with HBsAg (76.19 +/- 3.90)% and controls (62.37 +/- 4.24)%, P>0.01. The ability of DC after loading with HBcAg to induce T lymphocyte proliferation (34,326 +/- 3088 cpm) was significantly higher than that of DC after loading with HBsAg (20,306 +/- 2897 cpm) and controls (3454 +/- 409 cpm), P greater than 0.01. The amount of IL-12 in MLR of DC after loading with HBcAg was (348 +/- 42.8) ng/L, which was significantly higher than those of DC after loading with HBsAg (226 +/- 30.6) ng/L and controls (116 +/- 15.6) ng/L, P greater than 0.01.

<b>CONCLUSIONb>Human dendritic cell stimulated with HBcAg could more efficiently present antigen and induce specific T cell immune response.

Adult ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Cell Proliferation ; Dendritic Cells ; cytology ; immunology ; metabolism ; Female ; Flow Cytometry ; Hepatitis B Core Antigens ; immunology ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B, Chronic ; blood ; virology ; Humans ; Lymphocyte Activation ; immunology ; Male ; Middle Aged ; T-Lymphocytes ; cytology ; immunology ; Young Adult

To investigate the effects of human anti-BAFF scFv-Fc against the hsBAFF, ICR mice were randomly divided into six groups: control, hsBAFF (1 mg x kg(-1)), hsBAFF (1 mg x kg(-1)) + Ab (1 mg x kg(-1)), hsBAFF (1 mg x kg(-1)) + Ab (2 mg x kg(-1)), hsBAFF (1 mg x kg(-1)) + human IgG (1 mg x kg(-1)) and hsBAFF (1 mg x kg(-1)) + human IgG (2 mg x kg(-1)) groups. The effects of scFv-Fc administration on the proliferation of B lymphocytes were evaluated using an MTT assay. The titres of antibody in the serum and B lymphocytes differentiation were assessed by ELISA and flow cytometry, respectively. The results showed that administration of scFv-Fc to mice injected with hsBAFF significantly prevented human BAFF-induced increases in splenic B cell numbers and serum immunoglobulin levels. Furthermore, this fully human antibody would avoid inducing the human anti-mouse antibody (HAMA) response when used in humans. These findings suggest that the compact antibody may be useful in therapeutic or diagnostic application of the BAFF-associated autoimmune diseases in human.
Animals ; B-Cell Activating Factor ; immunology ; metabolism ; B-Lymphocytes ; cytology ; Body Weight ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Female ; Humans ; Immunoglobulin Fc Fragments ; immunology ; metabolism ; Immunoglobulin G ; blood ; immunology ; Immunoglobulin M ; blood ; Mice ; Mice, Inbred ICR ; Random Allocation ; Recombinant Fusion Proteins ; immunology ; metabolism ; Single-Chain Antibodies ; immunology ; metabolism ; Spleen ; cytology