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

The anatomical location of lymphocyte ontogeny in the developing mouse embryo remains controversial. To define the site that can generate lymphocytes de novo, the intraembryonic splanchnopleura (SP) and extraembryonic yolk sac (YS) at 8.5 days postcoitum, when systemic circulation is not established, were investigated. The results indicated that in standard colony forming assay, the cells from both splanchnopleura and yolk sac formed typical myeloerythroid colonies, but their types were distinct. When cocultured with the OP9, the splanchnopleura produced B cells expressing B220, CD19 and surface IgM. Using a three-step culture protocols with the OP9 expressing Delta-like 1 as feeders, the splanchnopleura produced immature T precursor cells (CD44-/CD25+) and more mature single positive T cells (CD4+/CD8-) after 16 days of incubation. However, the yolk sac failed to generate B and T lymphocytes under identical conditions. It is concluded that prior to linked embryonic circulation, the splanchnopleura other than the yolk sac had robust lymphoid potential in vitro. In the future, more reliable evidence from novel model animals will ultimately delineate the embryonic origin of lymphocytes in vivo.
Animals ; B-Lymphocytes ; cytology ; Cell Differentiation ; Coculture Techniques ; Female ; Hematopoietic Stem Cells ; cytology ; Mice ; Mice, Inbred C57BL ; Pregnancy ; T-Lymphocytes ; cytology ; Yolk Sac ; cytology

<b>OBJECTIVEb>To investigate the frequency distribution features of innate-like lymphocytes (iNKT cells, γΔT cells and B1 cells) in peripheral blood of normal adults.

<b>METHODSb>The flow cytometry with 6 fluorescence staining was used to detect the percentages of iNKT lymphocytes, γΔT lymphocytes, B1 lymphocytes and adaptive T lymphocyte, B2 lymphocytes in peripheral blood lymphocytes of 50 normal adults. The difference and correlation between these lymphocyte subsets were analyzed by statistical software.

<b>RESULTSb>The percentage of iNKT cells in peripheral blood of 50 normal adults was 0.18% (0.01%-2.01%), the percentage of γΔT cells was 4.90% (1.45%-20.14%), the percentage of B1 lymphocytes was 1.62% (0.20%-3.77%), the percentage of adaptive T cells was 63.52% (33.20%-83.22%), the percentage of B2 cells was 6.64% (3.07%-13.80%). B1 and B2 were two subsets of B lymphocyte, the percentage of B2 in B lymphocyte was 81.43% (57.90%-94.12%) and more than that of B1 lymphocyte; the percentage of B1 lymphocytes was 17.28% (5.28%-41.13%). In T lymphocyte group the percentage of iNKT cell was 0.32% (0.01%-3.6%), the percentages of γΔT cells and adaptive T cells were 7.55% (3.04%-27.66%) and 91.98% (72.22%-96.86%) respectively. Spearman correlation analysis was used to analyze the correlation between the percentages of several lymphocyte subsets. There was a positive correlation between iNK T cells and γΔT cells, γΔT cells and adaptive T cells, B1 cells and B2 cells (r=0.39, P=0.0056; r=0.6028, P<0.0001; r=0.4791, P=0.0004). It was also found that the percentage of iNKT cells in female peripheral blood lymphocytes was 0.29% (0.06%-2.01%), and significantly higher than that in male peripheral blood lymphocytes 0.12% (0.01%-1.37%) (P<0.05).

<b>CONCLUSIONb>The percentages of γΔT cells, B1 cells and iNKT cells in peripheral blood lymphocytes of normal adults are significantly lower than that of adaptive lymphocytes, and their contents in peripheral blood decrease in turn. There are no sex differences in the percentages of these lymphocyte subsets except iNKT cells.

Adult ; B-Lymphocytes ; cytology ; Female ; Flow Cytometry ; Humans ; Male ; Natural Killer T-Cells ; cytology ; T-Lymphocyte Subsets ; cytology

BACKGROUND: We investigated the characteristics of the mononuclear cells remaining in the leukoreduction system (LRS) chambers of Trima Accel(R) in comparison with those of standard buffy coat cells, and evaluated their potential for differentiation into dendritic cells. METHODS: Twenty-six LRS chambers of Trima Accel(R) were collected after platelet pheresis from healthy adults. Flow cytometric analysis for T, B, NK, and CD14+ cells was performed and the number of CD34+ cells was counted. Differentiation and maturation into dendritic cells were induced using CD14+ cells seperated via Magnetic cell sorting (MACS(R)) Seperation (Miltenyi Biotec Inc., USA). RESULTS: Total white blood cell (WBC) count in LRS chambers was 10.8x108 (range 7.7-18.0x108). The median values (range) of proportions of each cells were CD4+ T cell 29.6% (18.7-37.6), CD8+ T cell 27.7% (19.2-40.0), B cell 5.5% (2.2-12.1), NK cell 15.7% (13.7-19.9), and CD14+ cells 12.4% (8.6-32.3) respectively. Although total WBC count was significantly higher in the buffy coat (whole blood of 400 mL) than the LRS chambers, the numbers of lymphocytes and monocytes were not statistically different. The numbers of B cells and CD4+ cells were significantly higher in the buffy coat than the LRS chambers (P<0.05). The median value (range) of CD34+ cells obtained from the LRS chambers was 0.9x10(6) (0.2-2.6x10(6)). After 7 days of cytokine-supplemented culture, the CD14+ cells were successfully differentiated into dendritic cells. CONCLUSIONS: The mononuclear cells in LRS chambers of Trima Accel(R) are an excellent alternative source of viable and functional human blood cells, which can be used for research purposes.
Adult ; B-Lymphocytes/cytology/immunology ; CD4-Positive T-Lymphocytes/cytology/immunology ; CD8-Positive T-Lymphocytes/cytology/immunology ; Cell Differentiation ; Dendritic Cells/*cytology/immunology ; Flow Cytometry ; Humans ; Killer Cells, Natural/cytology/immunology ; Plateletpheresis/*instrumentation

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.
B-Lymphocytes ; immunology ; Dendritic Cells ; immunology ; Humans ; Killer Cells, Natural ; immunology ; Mesenchymal Stromal Cells ; cytology ; immunology ; T-Lymphocytes ; 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

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

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

<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>OBJECTIVEb>To investigate the predictive value of neutrophil/lymphocyte ratio (NLR) and platelet/lymphocyte ratio (PLR) for the patients with diffuse large B-cell lymphoma (DLBCL).

<b>METHODSb>The clinical data of 57 DLBCL patients admitted in the First Affiliated hospital of Anhui Medical University were analyzed retrospectively. According to ROC curve, the cut-off value for NLR and PLR was deterimined, and the patients were divided into high and low NLR/PLR groups before first chamotherapy. Then the relation of NLR and PLR with overall survival (OS) and progression-free survival (PFS) was analyzed by univariate and multivariate COX regression.

<b>RESULTSb>The optimal cut-off value for NLR and PLR was 2.915 and 270.27, respectively. NLR at the diagnosis was found to be an independent predictor for OS and PFS by univariate and multivariate analysis, while the PLR was an independent predictor for PFS, but did not affect the OS.

<b>CONCLUSIONb>NLR and PLR may provide additional prognostic information for DLBCL patients.

Blood Platelets ; cytology ; Disease-Free Survival ; Humans ; Lymphocyte Count ; Lymphocytes ; cytology ; Lymphoma, Large B-Cell, Diffuse ; diagnosis ; Multivariate Analysis ; Neutrophils ; cytology ; Prognosis ; Retrospective Studies