Preparation of lymphocytes from small volumes of peripheral mouse blood

Summary A method was developed which allowed for the isolation of lymphocytes (95% of leukocytes in the final suspensions) from small volumes of peripheral mouse blood (38% recovery). The method proved of value for studies of murine lymphocytes which required biochemical analysis of samples from individual animals on a serial basis.This work was supported in part by a University College Faculty Research Grant.     

Changes in the plasma membrane surface of lymphocytes stimulated in vivo with DNCB.

Stereological principles have been used to evaluate ultrastructural changes which accompany the transformation of lymphocytes stimulated in vivo with dinitrochlorobenzene (DNCB). Whereas unstimulated lymphocytes and blast cells have slightly more than the minimal containing plasmalemmal surface for their volume, stimulated (blast-derived) lymphocytes have an excess surface area in the order of 30%. This observation is discussed in the context of altered cell function and the biosynthesis of additional membrane components.     

Effect of taurine on mitogen response of human lymphocytes

Summary Taurine selectively inhibits the phytohemagglutinin-stimulated incorporation of³H-thymidine by human cultured lymphocytes (50% inhibition by 12.5 mM taurine). Decreasing effects of taurine on Na–K ATPase activity or on calcium accumulation by lymphocytes might be responsible for its action on cell proliferation.Thanks are due to Mr F. Cervantes Salas for technical assistance. This work was partly supported by grants. No. 5 RO1-EY 02540-02 from the National Eye Institute and No. PCCBNAL 790219 from CONACyT.     

Changes in the plasma membrane surface of lymphocytes stimulated in vivo with DNCB

Summary Stereological principles have been used to evaluate ultrastructural changes which accompany the transformation of lymphocytes stimulated in vivo with dinitrochlorobenzene (DNCB). Whereas unstimulated lymphocytes and blast cells have slightly more than the minimal containing plasmalemmal surface for their volume, stimulated (blast-derived) lymphocytes have an excess surface area in the order of 30%. This observation is discussed in the context of altered cell function and the biosynthesis of additional membrane components.We wish to thank Prof. R. Barer for his continued support and encouragement. The work was undertaken by M.M. A.-H. in part fulfilment of requirements for the degree of Ph. D. in this department. M.M. A-H. was supported by the Ministry of Higher Education and Scientific Research, Government of Iraq.     

Effect of taurine on mitogen response of human lymphocytes

Taurine selectively inhibits the phytohemagglutinin-stimulated incorporation of 3H-thymidine by human cultured lymphocytes (50% inhibition by 12.5 mM taurine). Decreasing effects of taurine on Na-K ATPase activity calcium accumulation by lymphocytes might be responsible for its action on cell proliferation.     

Kinetics of the appearance of HLA-DR antigens on human alloactivated T lymphocytes and the demonstration of new antigenic determinants

By studying serologically the appearance of HLA-DR determinants on T lymphocytes activated by a mixed lymphocyte culture, we have been able to demonstrate the existence of a new class of antigenic determinants distinct from classical HLA-DR antigens. Indeed, some monospecific anti-DR sera were cytotoxic from some alloactivated T cells, though not directed against their HLA-DR specificity. The absorption of these anti-sera on B lymphocytes bearing the HLA-DR antigen against which they were directed, did not remove their reactivity on alloactivated T lymphocytes. The absorption of the same anti-sera on activated T lymphocytes did not affect their anti-DR reactivity. This study shows the existence of new antigenic determinants expressed by T lymphocytes during their activation: alloactivated T lymphocyte antigens (AATL).     

Immigration of lymphoid precursor cells into the thymic rudiment inXenopus

Summary The origin of thymic lymphocytes was investigated, using a new reliable method to mark cells inXenopus. It was easily observed that extraneous cells immigrated into the thymic rudiment 4 days after fertilization and differentiated into a cell population identified as thymic lymphocytes in a fully developed thymus. Clearly, lymphoid precursor cells are of extrinsic origin.This work was supported in part by a grant from the Ministry of Education, Science and culture of Japan (No. 59770026).     

Sur le rôle joué par l'atrophie du thymus dans l'inhibition de la lymphopoïèse provoquée par la carence alimentaire en protéines chez le rat mâle

Summary In protein-depleted rats, atrophy of the thymus is partially responsible for the quantitative reduction of medium and large blood lymphocytes which precedes the decrease in the number of small lymphocytes.On the other hand, during recovery from protein starvation the restoration of medium and large lymphocytes is much more pronounced in intact than in thymectomized rats.     

Receptors for cytomegaloviruses on the surface of human lymphocytes]

The relationships between cytomegalovirus (CMV) and lymphocytes have already been noted because of: (a) the immunological abnormalities induced by this virus, and (b) activation of latent CMV in the course of lymphocyte reactions associated with anti-histocompatibility antigen immune response. The present work shows that the lymphocyte surface may have specific receptors for CMV. Cultured fibroblasts infected with DMV were incubated with lymphocytes isolated from the blood of human immune subjects. Rosettes defined by the adherence of three or more lymphocytes around a fibroblast were formed in infected preparation while no rosettes were seen with normal control fibroblasts. Approximately 1.2 per cent of lymphocytes were involved in the formation of these rosettes. Rosette formation is inhibited when infected fibroblasts have been incubated with anti-CMV antibodies prior to the addition of lymphocytes.     

Detection, by direct lymphocyte immunocytotoxicity, of antigens in cells transformed by Rous virus]

The hamster cells transformed by the Rous Sarcoma Virus (V.S.R.) evidenced surface antigenic alterations that were detected by a method of cellular mediated immunocytotoxicity. Immune hamster lymphocytes were added to tritiated proline prelabeled target cells. These lymphocytes were able to lyse specifically the V.S.R. transformed cells.     

运用磁珠分选法建立免疫细胞共培养体系

目的以相互作用的几种免疫细胞上共同表达的抗原物质为标志,运用磁珠分选技术,从慢性乙型肝炎患者外周血单个核细胞中同时获得并建立体外多细胞共培养体系,体外观察细胞之间的相互作用.方法采用梯度离心法分离30例慢性乙肝病毒感染者外周血单个核细胞(PBMC),采用免疫磁珠分选法分离获得 CXCR5+细胞,通过流式细胞仪检测 CXCR5+细胞纯度和组成,并将 CXCR5+细胞进行体外培养.在分离得到的 CXCR5+细胞的基础上,将 CXCR5+细胞分为3组,分别为对照组:不加任何刺激;实验组一:加入 IL 21刺激培养,实验组二:与人肝癌细胞系(HepG2)2215细胞混合培养.体外培养一周后,流式细胞仪检测实验组与对照组细胞 B细胞数量变化,ELISA测定培养体系中上清液 HBe抗体的含量.结果1)用流式细胞仪鉴定 CXCR5+细胞的纯度和构成:CXCR5+细胞纯度为85.5±5.8%,其中 Tfh细胞占23.8±7.4%,B细胞占35.6±7.6%;2)培养7天后,IL 21刺激组 B细胞百分率为47.2±1.8%,与(HepG2)2215混合培养组 B细胞百分率为40.2±3.5%,空白对照组 B细胞百分率为36.6±7.5%,IL 21刺激组与对照组,(HepG2)2215混合培养组与对照组 B细胞百分率均有显著差异(p<0.05);3)ELISA检测培养液上清 HBeAb定量,IL 21刺激组为0.668±0.094pg/ml,空白对照组为0.378±0.088pg/ml,两组有显著差异(p<0.05).结论使用间接免疫磁珠分离法可成功建立 CXCR5+B淋巴细胞和 Tfh细胞的共培养体系,为进一步体外研究细胞之间相互关系奠定基础     

Characterization and relative abundance of maxi-chloride channels in Epstein-Barr virus (EBV) producer: B95-8 cells

Several Epstein-Barr virus (EBV)-transformed cell lines were used to investigate the pathogenesis of lymphoproliferative diseases and nasopharyngeal carcinoma. The studies focus on the events occurring inside the membrane. On only one occasion, the cell membrane of EBV-transformed B lymphocytes from a cystic fibrosis patient was found to express defective Cl channels (CFTR; Cystic Fibrosis Transmembrane conductance Regulator), as in the airway epithelial cell. No other type of channel in EBV-transformed cells has so far been investigated. In this study, the cell membrane of the B95-8 cell was examined by the patch-clamp technique and compared to the non-EBV-infected BJAB cell. The high conductance (300 pS) maxi-chloride (Cl) channel activity was the most frequently observed event in inside-out configurations. Under similar experimental conditions, we have found a significantly higher probability of detecting maxi-Cl channel activity on the cell membrane of B95-8 cells (69%) than on BJAB cells (27%), or as previously reported on resting murine B lymphocytes (38%) or intact human T lymphocytes (37%). The relative abundance of the maxi-Cl channel on B95-8 cells may be linked to EBV infection and/or secretory ability.     

Transfer of genetic information from T to B human lymphocytes during an immune response to herpes simplex virus]

Human blood lymphocytes carrying different allotypes were divided into B and T subpopulations and cultured in presence or in absence of ultraviolet inactivated Herpes Simplex Virus. Isolated B or T cells did not produce any antiherpetic activity. The B lymphocytes cultured in presence of 1 or 50% of the supernatant collected from virus exposed T cells synthesized on antiherpetic antibody with some allotypic markers of the T cell donor.     

The environment in which lymphocytes differentiate influences their ability to cooperate in vivo

Summary Whether or not parental B lymphocytes cooperate with F1 T lymphocytes depends upon the environment in which parental bone marrow cells differentiate. Only those parental B lymphocytes that have differentiated in the F1 environment are able to cooperate with F1 T lymphocytes.Postdoctoral Investigator supported by Subcontract No. 3322 from the Biology Division of Oak Ridge National Laboratory to the University of Tennessee and by the Fulbright-Hays Program.Operated by the Union Carbide Corporation for the Department of Energy.     

Determination of trace elements in peripheral lymphocytes from tumor-bearing rats.

Neutron activation analysis of peripheral lymphocytes from healthy and Walker 256 carcinosarcoma-bearing rats resulted in the quantitative determination of Co, Cu, K, Mn, Rb, Se and Zn, and a qualitative determination of Au and Sb. From corresponding plasma samples, Co, Zn, Rb, Se and Cs were analyzed. Differences in trace element concentrations could only be detected in plasma of tumor-bearing animals which showed a decreased Zn and Co content of 50% and 30%, respectively, and a rise of Rb when compared to plasma of control animals.     

Somatostatin reduces the release of colony-stimulating activity (CSA) from PHA-activated mouse spleen lymphocytes.

PHA-activated lymphocytes release colony-stimulating activity (CSA) for macrophage-granulocyte precursor cells (colony forming units, CFUc) in the culture medium. Somatostatin, known to interfer with ribosomal protein synthesis, was demonstrated to reduce the release of CSA from PHA-treated mouse spleen lymphocytes.     

Inhibition of DNA biosynthesis in human lymphocytes by alpha 1-antitrypsin]

Incorporation of tritiated thymidine into human lymphocytes from tonsils is markedly inhibited by purified alpha1-antitrypsin-preparations. This inhibitory effect is observed in lymphocytes stimulated by a mitogen factor (phytohemagglutinin) as in non stimulated lymphocytes.     

Determination of trace elements in peripheral lymphocytes from tumor-bearing rats

Summary Neutron activation, analysis of peripheral lymphocytes from healthy and Walker 256 carcinosarcoma-bearing rats resulted in the quantitative determination of Co, Cu, K, Mn, Rb, Se and Zn, and a qualitative determination of Au and Sb. From corresponding plasma samples, Co, Zn, Rb, Se and Cs were analyzed. Differences in trace element concentrations could only be detected in plasma of tumor-bearing animals which showed a decreased Zn and Co content of 50% and 30%, respectively, and a rise of Rb when compared to plasma of control animals.     

A population of human lymphocytes staining for esterases

Summary A population of lymphocytes is found to stain positively for esterases. The positively staining lymphocytes are more predominant among T than B lymphocytes and are significantly increased on stimulation with PHA. Treatment with cholinestrase inhibitors reduces their number significantly.This work was supported by a grant from the Medical Research Council of Canada.