bluuebunny/crossref_metadata_2025_split huggingface dataset:
Oligoclonal Expansion of TCR V beta Subfamily T Cells in Patients with B-ALL.
Authors: Li Yangqiu , Huang Meijuan , Zhou Yubing , Chen Shaohua , Yang Lijian 
Published: October 2019

Abstract
Clonal expansion of TCR V beta subfamily T cells were identified in peripheral blood and tumor infiltrated tissues lymphocytes from some solid tumors and leukemias, which were thought to be relative to tumor associated antigen and be benefited for design of the strategy of specific immunotherapy. The aim of this study is to investigate the situation of TCR V beta gene repertoire and clonal expansion in peripheral blood T cells from patients with B-ALL. The complementarity determining region 3 (CDR3) of TCR V beta 24 subfamily genes in peripheral blood mononuclear cell from 13 cases with untreated B-ALL were amplified using RT-PCR. The PCR products were further labelled by fluorescein and then analyzed by genescan technique for detection of the CDR3 size, to determine the clonality of T cells. The results showed that only 2–18 V beta subfamilies were identified in 13 B-ALL cases respectively. The frequency expression of V beta subfamilies was V beat 13 (84.6%) and V beta 21 (76.9%). No positive products were detected in V beta 4, 6, 8, 18, 20 and V beta 24 subfamilies. Clonal expansive T cells in one or more V beta subfamilies were found in all 13 cases. The display frequency of clonal expansive T cells in V beta 21 (46.1%) and V beta 23 (30.8%) subfamilies were higher than others. In conclusions, the results indicated that the dominant utilization of TCR V beta repertoire could be found in peripheral blood T cells from patients with B-ALL, meanwhile clonal expansive T cells were existed. It may be a feature of the host immune response for leukemia associated antigen. The clonal expansive tendency of T cells in V beta 21 and V beta 23 subfamilies was rather obvious, which maybe correlated with certain malignant B-cell clone. Isolation and amplification of the clonal expanded V beta T cells in vitro may be necessary for identification of the specific cytotoxicity for leukemia cells.
Polyclonal IgG4 Hypergammaglobulinemia Associated with Lymphadenopathy and Renal Disease: A Novel Syndrome.
Authors: Boulanger Emmanuelle , Fuentes Vincent , Meignin Veronique , Mougenot Beatrice , Labaume Sylvaine , Cogne Michel , Aucouturier Pierre , Oksenhendler Eric , Clauvel Jean-Pierre , Ronco Pierre , Lassoued Kaiss 
Published: October 2019

Abstract
Marked polyclonal IgG4 hypergammaglobulinemia has exceptionally been reported. Here we report on two patients who presented an unique syndrome including a predominant and massive increase in serum levels of the IgG4 subclass. Clincal, biological and pathological data were described. Production of IgG4 by patients’ peripheral blood lymphocytes was studied, as well as the capacity of patients’ sera and T lymphocytes to induce IgG4 production by normal B cells. The possible involvement of IL4 and IL13 in the increased IgG4 production was also investigated. The two patients had anemia, lymphadenopathy, renal manifestations and a marked polyclonal IgG4 hypergammaglobulinemia leading to hyperviscosity syndrome in one case. The IgG4-producing cell percentage was significantly increased in both cases. By contrast with normal sera, both patients’ sera significantly increased the IgG4-producing cell percentagae when incubated with CD40-stimulated normal B lymphocytes. Similar effects were obtained with the culture supernatants of patients’ activated T cells. IL4 and IL13 serum concentrations were normal in the two patients. Anti-IL4 and/or anti-IL13 antibodies were unable to antagonize the IgG4 synthesis. The increased IgG4 production was found to be mediated by soluble, heat-resistant factor(s) that did not require the Stat6 signaling pathway. We described here a novel syndrome characterized by a significantly increased IgG4 production, mediated by soluble, heat-stable factor(s) most probably secreted by activated T cells, and distinct from IL4 and IL13.
Deficient IL-10 Secretion by Dendritic Cells (DCs) and Activated Lymphocytes in Aplastic Anemia (AA) Patients.
Authors: Archuadze Shorena , Mikhailova Elena , Sadovnikova Elena , Parovichnikova Elena , Savchenko Valeri 
Published: October 2019

Abstract
Background: Immunoregulatory cytokines (IL-10 and IL-12) secreted by the activated DCs and lymphocytes (LCs) mediate Th1 and Th2 immune response, respectively. Characteristic increase in Th1/Th2 ratio in AA patients might be responsible for autoreactive T-cell mediated suppression of hematopoiesis. However, functional characteristics of DCs initiating the Th1/Th2 disbalance in AA patients remain unelucidated. Aims: To study the functional characteristics of DCs and activated LCs in AA patients. Methods: We have carried out the flow cytometry analysis of the expression of costimulatory molecules by DCs generated from peripheral blood mononuclear cells (PBMC) of 13 AA patients before and after the immunosuppressive therapy (IST). DCs from 4 healthy donors (HD) were studied as a control group. Supernatants (SNs) of LPS activated DC cultures and LC cultures with or without PHA stimulation were studied for IL-10 production by ELISA. CD4+CD25+ regulatory T cell population of AA patients and HDs has being evaluated by means of magnetic beads for the negative selection of CD4+ T cells and immunofluorescence staining with anti-CD25 monoclonal antibodies. Results: DCs of 53,4% (7/13) of AA patients exhibited lower than HDs’ DCs expression of CD83 - marker of mature DCs. Level of IL-10 production by DCs from AA patients was significantly lower compared with HDs; opposite to HDs’ LCs, unstimulated LCs from 88,8% (16/18) of untreated AA patients failed to produce IL-10, however, LPS stimulation resulted in IL-10 secretion though to significantly lower levels compared with HDs; IST enhanced IL-10 production by DCs as well as stimulated and unstimulated LCs from AA patients, though to lower amounts than those of HDs. Few experiments in AA patients after long-term successful IST showed that DCs of AA patients with complete remission tend to produce even more IL-10 than those of HDs. Conclusion: Altered expression of CD83 by the DC from AA patients might influence the activation of cytotoxic T-cells. IL-10 downregulation might inhibit the generation of CD4+CD25+ regulatory T cell population and consequently, contribute to the expansion of cytotoxic T-cells and to the development of AA.
Rapid Detection of SAP Deficiency in Cytotoxic Lymphocytes from Patients with X-Linked Lymphoproliferative Disease and Their Family Members.
Authors: Tabata Yasuhiro , Villanueva Joyce , Lee Susan M. , Zhang Kejian , Kanegane Hirokazu , Miyawaki Toshio , Sumegi Janos , Filipovich Alexandra H. 
Published: October 2019

Abstract
Background: X-linked lymphoproliferative disease (XLP) is a primary immunodeficiency, which is characterized by an extreme susceptibility to Epstein-Barr virus (EBV). In half of the XLP patients, primary EBV infection can be fatal with explosive activation and proliferation of lymphocytes in many organs, which leads to fluminant hepatitis and bone marrow failure with hemophagocytosis. Mutations in the SH2D1A gene, which encodes the SAP protein, have been described in a proportion of patients with the clinical syndrome of XLP. The diagnosis of XLP is still difficult given its clinical heterogeneity, and the lack of a readily available rapid diagnostic laboratory test, particularly in cases without a family history of XLP. XLP should always be a consideration in males with EBV-associated hemophagocytic lymphohistiocytosis (EBV-HLH).
Methods: Four-color flow cytometric analysis was used to establish normal patterns of SAP expression for control subjects, and patterns of SAP staining in cytotoxic lymphocytes (CD8+ T cells, CD56+ T cells and natural killer [NK] cells). This assay was used to study patients with clinical syndromes consistent with XLP, and in some cases; their family members.
Results: 6 patients with XLP confirmed by detection of mutations in the coding regions of SH2D1A showed lack of intracellular SAP in all cytotoxic cell types; one was tested presymptomatically. Carriers of SH2D1A mutations tested (mother and sister) had decreased SAP staining patterns below normal levels. Eleven males with clinical syndromes consistent with XLP, predominantly EBV-HLH who were demonstrated to have normal or increased perforin expression were also studied. All patients showed normal SAP expression including one case with a convincing X-linked family history; all were subsequently shown to have no mutations in SH2D1A. Additionally, one asymptomatic adult male with an intronic mutation previously published as causing XLP was found to have normal SAP expression.
Conclusions: Four-color flow cytometry provides diagnostic information that may speed the identification of this fatal disease, differentiating it from other EBV-HLH. We also suspect it will prove useful in substantiating or excluding disease-causing genetic variants in the SH2D1A gene.
Driving T Cell Development in the Thymus.
Authors: Joao Cristina M. , Ogle Brenda M. , Cascalho Marilia , Platt Jeffrey L. 
Published: October 2019

Abstract
Background: Classic reports on lymphocyte development hold that B and T cells develop independently. This concept derives in part from the observation that patients with pure B cell immunodeficiency and hypogammaglobulinemia have a normal thymus and T cell numbers. Our recent findings however challenge this concept. We found that T cell development depends not only on the interaction of T cell precursors with thymic epithelial cells but also on other cells. Here we report that those other cells are B cells.
Aims: The purpose of this study was to determine whether B cells drive T cell development and TCR diversification in the thymus.
Methods: We compared the number of sub-populations of thymocytes and TCR repertoire diversity in B-cell deficient and B-cell proficient mice and in B cell deficient mice following immunoglobulin (Ig) injections. Total leucocytes numbers were determined with a Coulter counter and numbers of thymocytes sub-populations were calculated by flow cytometry analysis. TCR repertoire diversity was measured by a novel method based on hybridization of TCR Vβ specific cRNA on a gene chip platform.
Results: In B-cell deficient mice the number of thymocytes was four times reduced and TCR Vβ chain diversity was up to one million times lower compared with wild type mice. Numbers and diversity were restored by treatment of the mice with gamma globulin (see table).
Conclusions: T cell development and diversification is driven by B cells.
Mice Number of total thymocytes (mean ± standard deviation) p Value β V TCR diversity of thymocytes (median; min.-max.) p Value C57BL/6 (wild mice) 1.3 x 108 ± 5.1 x 107N=7 4.7 x 106; 1.0 x 105 − 1.1 x 108N=5 JH−/− (B cell immunodeficient mice) 3.1 x 107 ± 1.7 x107N=7 0.002 5.9 x 102; 3.6 x 102 − 1.1 x 103N=5 0.0002 JH−/− treated with Ig 3.9 x 107 ± 1.4 x106N=2 0.20 1.1 x 105; 2.7 x 100 − 7.7 x 105N=4 0.08
Structural Analysis of Janus Kinases.
Authors: Boggon Titus J. , Li Yiqun , Eck Michael J. 
Published: October 2019

Abstract
Janus Kinase 3 (Jak3) plays an essential role in hematopoietic signaling. Primarily expressed in B-, T- and Natural killer cells, it is activated through the γc chain of interleukin-2 like cytokine receptors (IL-2, -4, -7, -9, -15, -21). Deficiency of catalytically active Jak3 or disruption of the Jak3:IL-2γc interaction results in severe combined immunodeficiency (SCID). Jak3-deficient humans demonstrate defects restricted to the immune system, suggesting that selective inhibition of Jak3 catalytic activity or interruption of the Jak3:IL-2γc interaction are potentially exploitable strategies to achieve immunosuppression. Jak kinases contain four defined regions; a catalytically active carboxy-terminal kinase, a pseudo-kinase, a SH2-like region and a N-terminal Ferm domain. To date, no direct structural information has been reported for portions of any of the Jak family kinases. Structural studies are underway to define crystallographically the kinase domain of Jak3 and the Jak3-Ferm:IL-2γc interaction. Structural insights into the mechanism of Jak activation and routes to specific inhibition will be discussed.
Cytotoxic Activity of Cytokine-Induced Killer Cells Correlates with Expression of SAP and SLAM.
Authors: Mehrle Stefan , Frank Susanne , Schmidt Jan , Buchler Markus W. , Schmidt-Wolf Ingo G.H. , Marten Angela 
Published: October 2019

Abstract
SAP is a small protein, consisting of a single SH2 domain which is mutant in humans with X-linked lymphoproliferative disease. Patients with XLP are affected by fatal EBV infections and malignant B cell lymphomas. The increased risk for B cell lymphomas is suggested to result from impaired immunosurveillance of B cell proliferation by T cells. Here, we investigated the role of SLAM/SAP for activation of effector cells with cytotoxic activity (CIK cells), which are generated by unspecific stimulation of the T cell receptor and addition of exogenous IL-2 as described previously. The TCR activation on day +1 resulted not only in a short peak of activated cells, but activation continued and increased in combination with IL-2. We observed a striking peak of SLAM (Signaling Lymphocyte Activation Molecule) on day +6 in form of extracellular detectable CD150 as well as at the level of proteins and mRNA. Interestingly, the cytotoxic activity and the amount of SHP-2 protein showed a similar pattern as the parameters mentioned above but were shifted one day. Comparing these data for correlation, we observed a significant correlation between cytotoxic activity and CD150 expression pattern (P<0.001) and amount of SLAM protein (P<0.02) as well as between amount of SHP-2 protein and SLAM parameters (P<0.03). IL-10 secretion did not correlate with any of the parameters investigated. Secretion of Th1/Th2 cytokines was determined using a cytometric bead assay. There was no change in the amount of IL-6 and TNF-alpha. IL-4 was below the detection threshold and IL-2 could not be analyzed due to exogenous addition.
IFN-gamma levels increased during cultivation, peaked on day +3 and then remained at about 4ng/ml. IL-10 secretion started after stimulation of cells by anti-CD3, peaked on day +3 and then decreased continuously. Between day +6 and day +7, only 0.86 ± 0.02ng/ml/24hrs/3x106cells were secreted. In summary, activation of peripheral blood cells with agonistic anti-CD3 antibody and exogenous IL-2, as used for generation of CIK cells, results in significant SLAM and SAP activation five days after TCR stimulation. This peak correlates with cytotoxic activity against tumor cells. SLAM expression and binding by SAP seems to be important in the activation of cytotoxic effector cells.
The Effects of Chloride Channel Blockers on Platelet Cytoplasmic Free Calcium Concentration and Platelet Aggregation.
Authors: Yin Songmei , Chen Xiaolin , Nie Danian , Xie Shuangfeng , Ma Liping , Wang Xiujiu , Wu Yu-Dan , Li Yiqing , Feng Jianhong 
Published: October 2019

Abstract
Objective To explore the effects of chloride channels on the regulations of platelet cytoplasmic free calcium concentration ([Ca2+]i) and platelet aggregation (PAG).
Methods Platelet were separated freshly and then activated by thrombin; The chloride channel blockers 4,4′-diisothiocyano-2, 2′-disulfonic acid stilbene (DIDS) or niflumic acid (NFA), and calcium channel blockers 1-{β-[3-(4-methoxyphenyl)propoxy]- 4-methoxyphenethyl}- 1H - imidazole hydrochloride (SK&F96365) or Nifedipine were added to react with the activated platelets. The effects of each agent on platelet [Ca2+]i and PAG were detected. The combine effects and the interactions among chloride channel blockers (DIDS, NFA) and calcium channel blockers (SK&F96365, Nifedipine) were also investigated.
Results Both DIDS and NFA [the concentration were12.5, 25, 50, 100 and 200μmol•L−1 respectively] could inhibit the PAG induced by thrombin (1U/ml) and the effect was dose-dependent. Compared with the control, they had no significant effects on resting [Ca2+]i. Compare with the control group, DIDS (100μmol•L−1), SK&F96365 (100μmol•L−1) and Nifedipine (100μmol•L−1) could significantly reduce the PAG, Ca2+ release and Ca2+ influx activated by thrombin in platelet (P<0.05). DIDS (100μmol•L−1) and SK&F96365 (100μmol•L−1) could enhance each other’s effect on reducing the PAG, Ca2+ release and Ca2+ influx (P<0.05). DIDS (100μmol•L−1) and Nifedipine (100μmol•L−1) could enhance each other’s effect on reducing Ca2+ release (P<0.05). NFA (100μmol•L−1) and SK&F96365 (100μmol•L−1) could weaken each other’s effect on Ca2+ release (P<0.05). NFA (100μmol•L−1) and Nifedipine (100μmol•L−1) could weaken each other’s effect on PAG, Ca2+ release and Ca2+ influx activated by thrombin in platelet (P<0.05).
Conclusion The chloride channel blockers DIDS and NFA have no effect on the resting [Ca2+]i and the leak calcium influx of platelet. DIDS can inhibit the Ca2+ release, Ca2+ influx and PAG of platelet induced by thrombin, while NFA can only inhibit the Ca2+ release of platelet induced by thrombin. There are interactions between chloride channel blockers and calcium channel blockers in resting [Ca2+]i and PAG of platelet. The opening of chloride channel can influence the cellular calcium movement of platelet.
Variability of PECAM-1 Expression on Blood Cells.
Authors: Berken Melanie S. , Newman Debra K. , Newman Peter J. 
Published: October 2019

Abstract
PECAM-1 is a 130 kDa member of the Immunoglobulin gene superfamily that is expressed on circulating platelets, neutrophils, monocytes, and certain lymphocyte subpopulations. Deficiency of PECAM-1 in mice results in hyper-reactive platelets, B cells, and mast cells, compromised vascular integrity, and endothelial cells that are more susceptible to radiation-induced apoptosis in vivo. Whether or not PECAM-1 expression levels vary in the human population, and whether variable PECAM-1 expression renders one more or less susceptible to cardiovascular disease, autoimmune disorders, or sepsis has not been investigated to date. As a first step in examining these questions, we have developed a quantitative flow cytometry-based assay to determine the number of PECAM-1 molecules expressed on the surface of blood and vascular cells. Whole blood samples obtained from anonymous normal volunteer blood donors were stained with saturating amounts of fluorochrome-conjugated anti-PECAM-1 antibody, and relative fluorescence intensities were determined for platelets, neutrophils, monocytes, and lymphocytes by flow cytometry. Polystyrene microbeads coated with calibrated numbers of antibody binding sites were saturated with the same fluorochrome-conjugated anti-PECAM-1 antibody and used to create a standard curve that allowed us to relate relative fluorescence intensities to known numbers of antibody binding sites. By plotting the fluorescence intensity of anti-PECAM-1 antibody binding to each cell population against this standard curve, we were able to quantify the number of PECAM-1 molecules on the surface of each cell population. We found that PECAM-1 expression levels most often ranged between 5,000–12,000 molecules/platelet, 6,000–10,000/lymphocyte, 30,000–40,000/neutrophil, and 50,000–80,000/monocyte. Some individuals, however, expressed 2–5 times more PECAM-1 on a given blood cell subtype. Interestingly, individuals who expressed high levels of platelet PECAM-1 did not necessarily also express high levels of leukocyte PECAM-1, and vice versa. Since a number of naturally-occurring alleles of the PECAM-1 gene have recently been associated with the progression of atherosclerosis, myocardial infarction, and/or coronary artery disease, correlation of genetic variation within the PECAM-1 locus with its variable expression on the surface of blood and vascular cells may permit addition of PECAM-1 to the growing list of cell adhesion and signaling receptors whose expression is linked to bleeding and clotting disorders in humans.
Effects of Estrogen Replacement Therapies on Mouse Platelet Funcion.
Authors: Leng Xing-Hong , Zhang Wei , Nieswandt Bernhard , Lopez Jose A. , Bray Paul F. 
Published: October 2019

Abstract
Clinical trials of hormone replacement therapy (HRT) have demonstrated an increase in stroke and myocardial infarction despite the beneficial changes in LDL and HDL cholesterols, and it has been suggested that HRT may adversely affect thrombosis and/or inflammation. Platelets participate in both of these processes and play a central role in thrombosis formation in high shear arterial vasculature. The effects of HRT on platelet function have been controversially reported, probably due to different hormone regimes used (e.g. different estrogenic compounds, different dosages and routes of administration). We used a murine model of menopause to test the hypothesis that HRT alters platelet reactivity. Bilaterally ovariectomized female littermates were treated with various estrogen regimes or matching placebo for 21 days. Three estrogen replacement therapy (ERT) regimes were studied: 1) oral conjugated equine estrogen (CEE) vs. placebo (n=5 pairs), 2) oral 17-beta estradiol (E2) vs. placebo (n=4 pairs), and 3) subcutaneously implanted E2 (SQ E2) vs. placebo (n=8 pairs). Washed platelets were used to focus on intrinsic platelet reactivity in the absence of plasma coagulation factors. Platelet reactivity was assessed by flow cytometric quantification of agonist-induced fibrinogen binding. Oral CEE, but not oral E2 enhanced the platelet response to collagen-related-peptide (CRP) (P<0.001 and P=0.87 for oral CEE and oral E2, respectively). This enhancement of platelet function was agonist specific since neither oral CEE (P=0.862) nor oral E2 (P=0.917) affected the platelet response to thrombin. We also tested whether the route of hormone administration affected platelet function. SQ E2 induced a condition different from that of oral E2: the platelet response to thrombin was enhanced (P=0.028), but only to low (<10 mU/ml), not high concentrations of thrombin. In contrast to oral E2, SQ E2 caused a reduction in CRP-induced platelet fibrinogen binding (P=0.030). Platelet surface expression of GPVI was reduced in response to SQ E2 (P<0.001), and the level of GPVI correlated with CRP-induced platelet fibrinogen binding (P<0.0001). In conclusion: 1) the effects of different estrogen replacement therapies on platelet function differ significantly, perhaps explaining some of the conflicts in the literature, 2) the effects of ERT on platelet function are dependent upon the ERT preparations and routes, 3) the effects of ERT on platelet function are agonist and agonist dose-dependent, and 4) SQ E2 induces lower platelet GPVI expression, most likely explaining the reduced response to CRP. A potential practical consequence of our findings is that oral E2 may be less prothrombotic than either oral CEE or transdermal E2.