bluuebunny/crossref_metadata_2025_split huggingface dataset:
Interleukin 2 receptor expression and interleukin 2 production in exponentially growing T cells: major differences between in vivo and in vitro proliferating T lymphocytes
Authors: Rocha Benedita , Lembezat Marie‐Pierre , Freitas Antonio , Bandeira Antonio 
Published: March 2007

AbstractIn the present study we have assessed the growth requirements for in vivo proliferating mature T cells. For that purpose we have selected experimental approaches which allow the study of exponential growth in vivo of a major fraction of T cells, and make it possible to obtain large numbers of T cells in cycle. Two types of growing T cell populations were used: peripheral T lymphocytes, proliferating exponentially after transfer into syngeneic athymic nude mice, and activated T cells in lymph nodes of normal mice draining the site of oxazolone administration. The results obtained show that mature T cell growth in vivo is not accompanied by expression of high‐affinity interleukin 2 (IL2) receptor in the majority of activated cells, is not abrogated by in vivo administration of anti‐IL2 receptor antibodies or enhanced by the in vivo injection of recombinant IL2, and that in vivo growing T cells do not produce detectable amounts of IL2, as evaluated functionally by limiting dilution assays or the presence of IL2 mRNA, detected by Northern blots or in situ hybridization. The presented data thus indicate that the rules known to apply to T cell activation and proliferation in vitro differ from those used by in vivo growing T lymphocytes, at least in the two systems studied.
Human interleukin 6 and tumor necrosis factor α production studied at a single‐cell level
Authors: Andersson Ulf , Matsuda Tadashi 
Published: March 2007

AbstractIndividual peripheral blood mononuclear cells, which produced interleukin 6 (IL 6) or tumor necrosis factor α, (TNF α), were studied by cytokine‐specific polyclonal or monoclonal antibodies (mAb) and immunofluorescence technique with UV microscopy. Lipopolysaccharide (LPS) induced IL 6 as well as TNF α production in the majority of the monocytes, but not at all in lymphocytes. Approximately every second monocyte made TNF α in response to LPS within 0.5 h from start of the cultures, when no IL6 or TNF α production occurred. The maximal number of TNF α‐synthesizing monocytes was observed 1.5 h later and then rapidly declined. LPS stimulation led to optimal IL 6 production 3 h after initiation of the cultures, with 90% of the monocytes expressing intracellular IL 6. LPS‐induced IL 6 synthesis started about 1 h after that of TNF α. Polyclonal T cell activation with staphylococcal enterotoxin A or anti‐CD3 mAb induced a biphasic production pattern of IL 6 as well as TNF α. Early IL 6 synthesis, which peaked 6–8 h from start of the cultures, occurred exclusively in monocytes, while late IL 6 production at 48 h was restricted to a small fraction of lymphoid cells. T cell mitogen induced early TNF α production, which peaked at 6 h, took mainly place in monocytes and to a minor degree in CD4+ as well as CD8+ T cells. The majority of the TNF α‐producing mononuclear cells at 24 h were of the CD4+ T cell lineage in the staphylococcal enterotoxin A‐ or anti‐CD3 mAb‐activated cultures. IL 6 as well as TNF α accumulated in the Golgi system, which resulted in a characteristic morphology of the staining, eliminating problems with evaluation of background signals.
Influence of sulfated carbohydrates on the accessibility of CD4 and other CD molecules on the cell surface and implications for human immunodeficiency virus infection
Authors: Thiele Bernhard , Braig Henk Ronald , Ehm Ingo , Kunze Rudolf , Ruf Bernhard 
Published: March 2007

AbstractThe modulatory activity of dextran sulfate with a relative molecular mass of 8 and 500 kDa and pentosan polysulfate with a relative molecular mass of 6 kDa on human T cell surface molecules CD2, CD3, CD4, CD8 and HLA‐DR was investigated by analytical flow cytometry. The 500‐kDa dextran sulfate induces a complete disappearance of the CD4 and a 50% diminuition of CD2 immune reactivity on peripheral blood lymphocytes after a 4‐h incubation while the low molecular mass polyanions do not. This modulation of the CD4 immune reactivity includes all CD4 epitopes investigated. It does not correlate with the antiviral effect of polyanions against human immunodeficiency virus infection. The interaction of polyanions with the CD4 presentation is temperature dependent and differs between fresh lymphocytes and immortal cell lines. From our data it can be concluded that mechanisms other than cell surface effects are responsible for the antiviral potency of these drugs. Implications for the modes of antiviral action of sulfated carbohydrates are discussed.
Characterization of cytoplasmic and nuclear polypeptides induced by interferon‐γ in a murine pre‐B cell leukemia
Authors: Franco Antonella , Fizzotti Marco , Gariglio Marisa , Gribaudo Giorgio , Cavallo Giorgio , Landolfo Santo 
Published: March 2007

AbstractThe effects of interferon‐γ (IFN‐γ) on protein synthesis of a pre‐B cell leukemia, L1210, have been studied by two‐dimensional gel electrophoresis. In total cell extracts, at least ten proteins were induced de novo, or increased in their expression, after an 18‐h IFN‐γ treatment, whereas in the nuclear extracts eight proteins were specifically induced. Of these, increased synthesis of a 40‐kDa/pI 5.9 cytoplasmic protein was the most prominent and reproducible. Most of these proteins appear to be specific for a defined step of differentiation, since they are not found in other B cell leukemias upon IFN‐γ treatment. Others appear to be tissue specific, since they are not induced in fibroblasts nor in T cells. In addition, synthesis of some of the induced proteins appeared to require rapid transcription of new mRNA, because actinomycin D markedly inhibited their formation when added immediately before IFN‐γ. In keeping with this finding, in vitro translation of mRNA from IFN‐γ‐treated L1210 cells into a rabbit reticulocyte lysate system, followed by analysis of the labeled proteins by two‐dimensional gel electrophoresis, revealed the appearance of at least seven proteins.Taken as a whole, these results demonstrate that in leukemic pre‐B cells IFN‐γ induces the transcriptional activation of genes coding for cytoplasmic and nuclear proteins, some of which could be employed as specific cell activation markers.
Morphologic and functional characterization of human peripheral blood T cells expressing the T cell receptor γ/δ*
Authors: Ferrini Silvano , Zarcone Daniela , Viale Maurizio , Cerruti Giannamaria , Millo Romano , Moretta Alessandro , Grossi Carlo E. 
Published: March 2007

AbstractThe morphologic and functional characteristics of cells freshly isolated from human peripheral blood and bearing a T cell receptor (TcR) γ/δ were analyzed. Cell preparations highly enriched for TcR γ/δ+ cells were obtained by treatment of E rosette‐forming lymphocytes with anti‐CD4 and anti‐CD8 monoclonal antibodies (mAb) and complement. These preparations consisted of 64–82% TcR γ/δ+ lymphocytes, as indicated by the sum of cells reacting with the BB3 and A13 mAb which define two distinct, nonoverlapping, TcR γ/δ+ cell subsets in the peripheral blood. TcR γ/δ cells were able to form conjugates with the natural killer‐sensitive K‐562 and with the natural killer‐resistant HL‐60‐R tumor cell lines. The cytochemical localization of lysosomal acid hydrolases showed that 95%–98% of the cells in the TcR γ/δ+ preparations had the morphologic features of granular lymphocytes. Moreover, electron microscopy analyses showed that TcR γ/δ+ cells had electron‐dense granules dispersed in the cytoplasm and a variety of smooth vesicles, a morphology identical to that of other CD3‐ or CD3+ granular lymphocyte subsets. Freshly isolated TcR γ/δ+ cells were unable to lyse K‐562 and natural killer‐resistant targets, such as HL‐60‐R and P815. However, low levels of target cell lysis were observed upon triggering of the effectors by anti‐CD3 TcR mAb or by lectin. After short‐term culture with inter‐leukin 2, TcR γ/δ+ cells acquired a strong cytolytic activity against K‐562 and HL‐60‐R target cells in the absence of triggering stimuli, and also displayed high levels of cytolytic activity against P815 in the presence of anti‐CD3/TcR mAb.
The chemotactic activity for granulocytes produced by virally infected fibroblasts is identical to monocyte‐derived interleukin 8
Authors: Damme Jo Van , Decock Benny , Conings Renè , Lenaerts Jean‐Pierre , Opdenakker Ghislain , Billiau Alfons 
Published: March 2007

AbstractSo far, the role of fibroblasts in inflammatory processes has been underestimated. We have previously shown that stimulation of fibroblasts with viruses or bacteria results in a simultaneous production of several cytokines, including interferon‐β, interleukin (IL) 6 and colony‐stimulating factors. We here report that virally infected fibroblasts produce also a chemotactic factor for granulocytes. The activity is inducible not only by measles virus but also by IL 1β and the double‐stranded RNA poly(rI) ˙ poly(rC). This factor, when purified to homogeneity, occurs as a 6–7‐kDa protein doublet upon sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. The pure protein is serologically related to a fully characterized granulocyte chemotactic peptide (GCP) from monocytes, designated IL8. Furthermore, the chemotactic factor from fibroblasts has an NH2‐terminal sequence identical to that of GCP/IL 8, small differences in NH2‐terminal processing being observed. Finally, in addition to diploid fibroblasts, the osteosarcoma MG‐63 cell line is also a producer of GCP/IL 8. It can thus be concluded that GCP/IL8 can be produced by several cell types in response to infection and that fibroblasts can contribute to chemotaxis in inflammation.
Ontogenic development of autoantibody repertoires in spleen and peritoneal cavity of normal mice: examples of T cell‐dependent and ‐independent reactivities
Authors: Huetz François , Poncet Pascal , Coutinho António , Portnoi Denis 
Published: March 2007

AbstractThe ontogenic development of B cell clonal precursors (BCP) reactive to bromelaintreated, syngeneic erythrocytes (BrMRC) and to single‐stranded DNA has been studied by limiting dilution of both spleen and peritoneal cells. It was found that the frequency of anti‐BrMRC BCP in the spleen is very low up to 4 weeks of age and slowly increases thereafter, to reach adult levels by 6–10 weeks. In the peritoneal cavity, no such BCP can be found before 2 weeks, but they occur at a very high frequency already by 3 weeks of age. Injection of adult, normal syngeneic T cells at birth has no apparent effect on the representation of anti‐BrMRC BCP in the peritoneal cavity, but brings these to adult levels or even higher in the spleen already at 3 weeks of age. Accordingly, adult athymic (nude) mice contain normal frequencies of BrMRC‐specific BCP in the peritoneal cavity but are devoid of such clones in the spleen. In contrast, the frequency of anti‐DNA BCP is very high throughout postnatal development in both spleen and peritoneal cavity, of normal and athymic mice, in both resting and naturally activated splenic B cell compartments, and it is independent of T cell transfers into nude animals. These results indicate the role of T cells in the establishment of some clonal specificities in the adult, splenic autoreactive B cell repertoire.
A novel CD3‐ J11d+ subset of CD4+CD8‐ cells repopulating thymus in radiation bone marrow chimeras
Authors: Matsumoto Koji , Yoshikai Yasunobu , Matsuzaki Goro , Asano Taiji , Nomoto Kikuo 
Published: March 2007

AbstractSequential appearance of T cell subpopulations occurs in the thymus of irradiated AKR (H‐2k, Thy‐1.1) mice at an early stage after transplantation with bone marrow cells of C3H/HeN (H‐2k, Thy‐1.2) mice. The donor‐derived thymocytes were first detected on day 8 after bone marrow reconstitution. Although most of the thymocytes were CD4‐CD8‐ cells, an appreciable level of CD4+CD8+ cells was detected in the thymus at this stage. The early appearing CD4+CD8‐ cells were a novel subset of thymocytes that were J11d+CD3‐. From day 10 to day 21 the proportion of CD4+CD8‐CD3‐J11d+ cells decreased while the proportion of CD4+CD8‐ cells and CD4+CD8‐CD3+J11d‐ cells increased. The CD4+CD8‐CD3‐ cells seem to diversify to form CD4+CD8+ thymocytes after short‐term culture in vitro. These results suggested the existence of a differential pathway from CD4‐CD8‐ cells to CD4+CD8+ cells via CD4+CD8‐ cells in the thymus.
Cell surface antigen CD5 is a marker for activated human B cells
Authors: Werner‐Favre Christiane , Vischer Thomas L. , Wohlwend Dominique , Zubler Rudolf H. 
Published: March 2007

AbstractA minor subset of B cells which in vivo express the surface antigen CD5, has attracted much attention because of its involvement in autoimmune responses. On the basis of observations showing self‐renewal capacity of such cells in mice and also the absence of a substantial change of CD5 phenotype during B cell activation in vitro, the CD5+ B cells are now generally considered to represent a separate cell lineage. In the present study, CD5‐ B cells were isolated by cell sorter and then stimulated in vitro with mutagenized EL4 thymoma cells in the presence of T cell supernatant. About 70% of the B cells were CD5+ after 3 days. Thus, the CD5 antigen behaves as a B cell activation marker. In our system we found that the frequency of rheumatoid factor‐producing B cells was on average three times higher in CD5+ than in CD5‐ B cells isolated ex vivo from human peripheral blood. Most likely this reflects frequent activation of such autoreactive B cells in vivo.
Analysis of HLA antigens on germ cells in human semen
Authors: Jassim Ali , Ollier William , Payne Annette , Biro Andrew , Oliver R. T. D. , Festenstein Hilliard 
Published: March 2007

AbstractNucleated cells other than sperm (NCOS) were isolated from human semen by centrifugation on a Ficoll density gradient. Using tissue‐specific monoclonal antibodies (mAb) > 99% of the NCOS were found to be sperm cell precursors (SpP). These cells were tested for the expression of class I and II (DR, DP and DQ) HLA antigens by using specific mAb. The anti‐HLA class I and II and anti‐β2‐microglobulin mAb reacted with < 1% of the NCOS. This was demonstrated by indirect immunofluorescence microscopy and fluorescence‐activated cell sorter analysis. These results were similar to those obtained from testing germ cells in frozen sections of normal adult testis using the same panel of mAb. In mixed lymphocyte‐NCOS cultures, the SpP failed to stimulate allogeneic lymphocytes even when different concentrations of cells were used. These results indicate little or no expression of HLA class I and II including the HLA‐D (T cell‐defined) determinant on the SpP, a phenomenon which could be of biological importance.
Intracellular events associated with inhibition of B cell activation by monoclonal antibodies to HLA class II antigens
Authors: Holte Harald , Blomhoff Heidi Kiil , Beiske Klaus , Funderud Steinar , Torjesen Peter , Gaudernack Gustav , Stokke Trond , Smeland Erlend B. 
Published: March 2007

AbstractWe have investigated several aspects of the inhibitory effects of monoclonal antibodies (mAb) directed against MHC class II antigens in B cell activation/proliferation, using a panel of mAb specifically reactive with antigens encoded by HLA class II loci (DP, DQ, DR). All mAb except the anti‐DP mAb inhibited significantly anti‐μ plus B cell growth factor‐induced DNA synthesis. Only one mAb, however, which was reactive with gene products of all three class II loci (DP, DQ, DR) inhibited anti‐μ‐induced DNA synthesis as well as c‐myc mRNA expression. In addition, the same mAb inhibited the early events induced by anti‐μ stimulation alone, including phosphatidylinositol turnover and elevation of [Ca2+]i. In contrast to previous findings in the murine system, none of the anti‐MHC class II mAb used in this study increased the cAMP levels.
Macrophage activation by interferon‐γ from host‐protective T cells is inhibited by interleukin (IL) 3 and IL 4 produced by disease‐promoting T cells in leishmaniasis
Authors: Liew Foo Y. , Millott Stephanie , Li Yun , Lelchuk Rosalia , Chan W. Ling , Ziltener Herman 
Published: March 2007

AbstractBALB/c mice are highly susceptible to Leishmania major infection. They develop a progressive fatal disseminating disease even with a minimum infecting dose. However, these mice are able to contain the disease if they are exposed to sublethal γ‐irradiation shortly before infection. Earlier studies demonstrated that CD4+ T cells from mice which had recovered from infection (Tr) can adoptively transfer resistance. In contrast, CD4+ cells from mice with progressive disease (Ts) not only failed to protect, but can reverse the protective effect of the Tr cells.Spleen cells from BALB/c mice which had recovered from L. major infection or which had progressive disease were cultured with leishmanial antigens in vitro. The culture supernatant from spleen cells of recovered mice (TrSN) contains high levels of macrophage‐activating factor (MAF) activity which can activate peritoneal macrophages to kill 51Cr‐labeled P815 cells and to eliminate intracellular parasites as measured by the reduction in [3H]thymidine uptake by residual parasites released from macrophages following sodium dodecyl sulfate treatment. The MAF activity of TrSN parallels that of recombinant interferon‐γ (IFN‐γ). In contrast, culture supernatant of spleen cells from mice with progressive disease (TsSN) contains no detectable MAF but it is able to neutralize the MAF activity of TrSN. The MAF‐inhibiting function of TsSN appears to be mediated by interleukin (IL) 3 and IL 4, since the MAF activity of TrSN and rIFN‐γ also can be inhibited by the addition of rIL 3 and rIL 4 but not by rIL 1 or rIL 2. Furthrmore, the MAF‐inhibiting activity of TsSN can be partially reversed by the addition of specific anti‐IL 3 or anti‐IL 4, but completely reversed by the combination of the two antibodies in vitro. These findings provide a mechanism for the immune regulation in leishmaniasis and a means by which the two subsets of CD4+ T cells influence each other through their modulation of macrophage function.
The VH gene sequences of anti‐DNA antibodies in two different strains of lupus‐prone mice are highly related
Authors: Eilat Dan , Hochberg Malka , Tron François , Jacob Laurent , Bach Jean‐François 
Published: March 2007

AbstractThe heavy and light chain V region sequences of an IgG anti‐DNA autoantibody (PME77), derived from a lupus‐prone (NZB x NZW)F1 mouse have been determined by mRNA sequencing. The Vx gene segment belongs to the Vx1A gene subgroup and is found in several (NZB x NZW)F1 and MRL lpr/lpr anti‐DNA antibodies, as well as in other antibodies of unrelated specificities. The VH gene segment appears to represent a unique gene or a subfamily of the large J558 VH gene family of the mouse, and is highly related to a germ‐line sequence of a major anti‐DNA idiotype (H130, IgM) of MRL mice. This anti‐DNA‐related VH segment has not been found, so far, to be expressed in antibodies with specificities for external or synthetic antigens; therefore, expression of such specificities may be regulated by powerful mechanisms of self tolerance in the healthy animal. In addition, both the heavy and light chain of the PME77 IgG antibody were found to contain somatic point mutations with a high ratio of replacement to silent mutations in complementarity determining regions. This IgM to IgG sequence relationship suggests an affinity maturation process, which is driven by the autoantigen.
Detection of perforin and granzyme A mRNA in infiltrating cells during infection of mice with lymphocytic choriomeningitis virus
Authors: Müller Christoph , Kägi David , Aebischer Toni , Odermatt Bernhard , Held Werner , Podack Eckhard R. , Zinkernagel Rolf M. , Hengartner Hans 
Published: March 2007

AbstractThe analysis of gene expression in cytotoxic T cells by in situ hybridization of serial liver and brain sections from mice infected with lymphocytic choriomeningitis virus (LCMV) and immunostaining with T cell marker‐ and virus‐specific antibodies revealed a close histological association of infiltrating lymphocytes expressing the perforin and granzyme A genes with virally infected cells. Maximal frequency of perforin and granzyme A mRNA‐containing cells on liver sections preceded by about 2 days maximal LCMV‐specific cytotoxicity of the lymphoid liver infiltrating cells. These results are most consistent with an involvement of perforin and granzyme A in cell‐mediated cytotoxicity in vivo.
Expression and ontogeny of murine CD2
Authors: Sen Jyoti , Arceci Robert J. , Jones William , Burakoff Steven J. 
Published: March 2007

AbstractCD2 was first defined as the erythrocyte rosetting protein on the surface of human T cells. Recently, the rat and murine homologues have been identified by cDNA cloning. In this report we demonstrate that CD2 is expressed on the surface of most adult murine peripheral lymphocytes and thymocytes by indirect immunofluorescence using an anti‐murine CD2 antiserum. The expression of CD2 on murine B cells was unexpected since in rat and human species it has been defined as a T cell‐specific marker. Furthermore, CD2 appears very early on fetal thymocytes during development. The level of surface expression increases from day 13 of gestation to day 17, after which the surface density appears to reach a steady state. Thus, CD2 is expressed on day‐13 thymocytes at the same stage that Thy‐1, Pgp1 and the TcRγ/δ/CD3 complex have been shown to be expressed.
Thymocyte development: an analysis of T cell receptor gene expression in 519 newborn thymocyte hybridomas
Authors: Happ Mary Pat , Palmer Ed 
Published: March 2007

AbstractWe have examined the frequency of expression of individual α and β chain V gene families in a population of immature T cells that has not been selected or tolerized. To accomplish this, we generated 519 T cell hybridomas from freshly isolated thymocytes of newborn C57BL/10 mice and subjected RNA from these hybrids to hybridization analysis with 11 Vα, 16 Vβ, Cγ and Cδ probes. Comparison of the expressed repertoire of Vβ gene segments in this newborn thymocte population with similar data previously generated from adult peripheral T cells (Bill et al., Proc. Natl. Acad. Sci. USA 1988. 85: 9184.) revealed two Vβ genes, Vβ12 and Vβ15, whose expression is decreased in the periphery possibly due to the effects of tolerance. An additional Vβ gene segment (Vβ10) and a Vβ gene family (Vβ5.1, Vβ5.2) were expressed more frequently in the mature, peripheral population than in the newborn thymus. These findings may represent two instances of positive selection of T cells. Furthermore, unlike VH gene segments, Dβ‐proximal Vβ genes are not overrepresented on this collection of immature thymocyte hybridomas. A similar analysis of Vα gene family expression was hampered by the fact that Vα gene segments were expressed in only 15% of newborn thymocyte hybridomas (compared to 58% of adult hybridomas). An unexpectedly large fraction (57%) of those newborn hybrids expressing a Vα gene segment was also found to express Cδ mRNA and further examination revealed that several Vα gene probes were actually detecting δ chain mRNA. The most notable of these was the Vα7 gene family, which accounted for approximately one‐third of the expressed Vα genes but was expressed exclusively as part of a γ chain mRNA. We found no examples of hybridomas co‐expressing both full‐length β and δ chain mRNA, despite significant numbers of hybridomas co‐expressing full‐length β and γ chain transcripts. This observation suggests that a large number of mature α/β cells may have proceeded through development without having gone through a stage where a δ chain message is expressed, thereby precluding their development into γ/δ cells.