bluuebunny/crossref_metadata_2025_split huggingface dataset:
Dendritic cells of the mouse recognized by two monoclonal antibodies
Authors: Breel Matthijs , Mebius Reina E. , Kraal Georg 
Published: March 2007

AbstractA new monoclonal antibody, MIDC‐8, is described which shows a comparable tissue distribution as the recently described NLDC‐145 antibody. It recognizes interdigitating cells, veiled cells and Langerhans cells in lymphoid organs and the skin of the mouse. In contrast to NLDC‐145 it recognizes a cytoplasmic component of these cell types. Its distribution is more restricted to the T cell‐dependent areas than NLDC‐145. Isolation of dendritic cells from lymph nodes, spleen and thymus revealed that both antibodies react with the in vitro isolated dendritic cells. The results show that these antibodies can be used to study dendritic cells in vitro and emphasize the relationship between the in vivo interdigitating cells of the T cell areas and the in vitro isolated dendritic cells.
Three‐cell‐type clusters of T cells with antigen‐presenting cells best explain the epitope linkage and noncognate requirements of the in vivo cytolytic response
Authors: Mitchison N. Avrion , O'Malley Christine 
Published: March 2007

AbstractUsing an established i.p. adoptive transfer system in which primed precursors of cytolytic (Tc) cells are combined with primed helper (Th) cells and alloantigen, the three‐cell‐type hypothesis of T‐T cooperation has been tested. This hypothesis assumes that cooperation takes place through the formation of clusters, consisting of one or more Tc precursors plus one or more Th cells binding to antigen on the surface of antigen‐presenting cells (APC). By using antigens in which the H‐2Db epitopes recognized by the Tc precursors are presented either on the same cell as the BALB minor histocompatibility epitopes recognized by the Th cells, or a different one, the value of epitope‐linkage has been explored. Epitope‐linkage is found to enhance the response at low concentrations of antigen but not at high ones, in accordance with the prediction that at low concentrations individual APC will usually take up either one antigen or the other but not both from an unlinked mixture, while at high concentrations most APC will take up both whether they are linked or not. In a further test of this hypothesis, no requirement for a cognate T‐T interaction could be detected.With the three‐cell‐type interaction thus better established, its implications in terms of efficiency compared with linked‐cognate interactions and its evolution are discussed.
Induction of T cell proliferation with anti‐CD3 switch‐variant monoclonal antibodies: effects of heavy chain isotype in monocyte‐dependent systems
Authors: Van Lier René A. W. , Boot Johan H. A. , De Groot Els R. , Aarden Lucien A. 
Published: March 2007

AbstractMonoclonal antibodies (mAb) directed against the CD3 (T3) antigen are able to induce proliferation in resting human T lymphocytes. T cell proliferation only occurs in the presence of monocytes that carry the proper Fc receptor for the mAb used. To further analyze the role of the Fc portion of anti‐CD3 mAb in proliferation induction, we isolated, starting from a γ1 anti‐CD3‐producing hybridoma, four heavy‐chain isotype switch‐variant antibody‐secreting clones, producing γ2b, γ2a, ϵ and α, respectively. All variant antibodies recognize the CD3 antigen as determined by immunoprecipitation and cross‐blocking experiments. With this series of isotype variant antibodies we were able, in proliferation induction experiments, to confirm the Fc receptor polymorphism for murine IgG2a, IgG2b and IgG1 on human monocytes. Moreover, we found that all 30 donors tested responded to the IgE anti‐CD3 antibody, while no IgA responders could be identified. The induction of proliferation by the IgE variant antibody does not require the 72‐kDa Fc receptor which is responsible for the interaction with mouse IgG2a. Nonresponsiveness to the IgG1 antibody, but not to the IgG2b or IgA variant antibodies, could be overcome by the addition of exogenous interleukin 2 to the cultures. When the switch‐variant antibodies were used to induce IgM synthesis in peripheral blood mononuclear cells only low IgM synthesis was found, with the exception of the IgE variant, which induced excellent T cell help for IgM production.
Activation of resting T lymphocytes by cross‐linked anti‐CD3 (T3)
Authors: Heumann Didier , Vischer Thomas L. 
Published: March 2007

AbstractIn this study we have examined the role of small numbers of adherent cells in the stimulation of highly purified resting T lymphocytes by cross‐linked monoclonal anti‐CD3 antibodies (T3). T cells were obtained by 3 cycles of purification, using adherence to plastic surface, nylon wool column separation and rosetting with 2‐aminoethyl isothiouronium bromide‐treated sheep red blood cells. They were stimulated either with T3 antibodies or with solid‐phase rabbit anti‐mouse IgG‐T3 complexes. In standard high density cultures (1 × 105−2 × 105 cells), soluble T3, interleukin (IL) 1 and IL 2, used separately, did not induce proliferation of T cells. Soluble T3 together with IL 2 slightly activated T cells to proliferate. Rabbit anti‐mouse IgG‐T3 complexes attached to the plastic wells induced stimulation in some cultures. The response was markedly increased after addition of IL 2, but not IL 1. The irregular response of these cultures suggested the presence of another variable signal. When cell numbers were reduced (12 × 103 cells), neither cross‐linking of the CD3‐Ti complex nor addition of exogenous IL 1 or IL 2, alone or in combination, stimulated the T cells to increase DNA synthesis. A positive response, comparable to complete peripheral blood mononuclear cells, was restored with 0.5% supplemented adherent cells, provided that IL 2 was present.
Rapid decrease in lymphocyte adherence to high endothelial venules in lymph nodes deprived of afferent lymphatic vessels
Authors: Hendriks Hans R. , Duijvestijn Adrian M. , Kraal Georg 
Published: March 2007

AbstractOcclusion of the afferent lymph flow to the lymph node (LN) results in both flattening of the endothelium of high endothelial venules (HEV) and a severe decrease in numbers of lymphocytes in transit across the walls of the flattened HEV. In the present study we have used the in vitro lymphocyte‐binding assay to investigate the ability of HEV in rat LN to bind lymphocytes at various time points after occlusion of the afferent lymph flow. In addition the specificity of T and B lymphocyte adherence to HEV of such operated LN was studied.In normal LN, lymphocytes adhered to virtually all HEV using the in vitro binding assay. However, 1 and 2 weeks after operation lymphocytes bound to only 50–60% of the HEV and by 3–6 weeks 20–30%. The total numbers of lymphocytes bound to these HEV had also diminished to 10% of the control value 3–6 weeks after operation. Morphometric analysis showed that this was not only due to a reduction in the area of HEV endothelium available for lymphocyte adherence by flattening of the high endothelial cells, but also to a strong decrease in the numbers of bound lymphocytes per unit area high endothelium. In spite of the reduction in numbers of adhering lymphocytes the T/B cell ratio did not change. The results show that the reduction in lymphocyte binding of HEV in operated LN is a rapid event, probably due to loss of high endothelial cell determinants involved in binding of lymphocytes. The decrease in lymphocyte binding clearly precedes flattening of HEV endothelium suggesting that the height of high endothelial cells is of secondary importance to lymphocyte adherence.
Bovine cytotoxic T cell clones which recognize lymphoblasts infected with two antigenically different stocks of the protozoan parasite Theileria parva
Authors: Morrison W. Ivan , Goddeeris Bruno M. , Teale Alan J. 
Published: March 2007

AbstractThe Muguga and Marikebuni stocks of Theileria parva differ on the basis of cross‐protection and in their schizont antigen profile determined with a panel of parasitespecific monoclonal antibodies. The phenotype and specificity of six cytotoxic T cell clones generated from an animal immunized against T. parva (Marikebuni) were investigated. All six clones had the BoT2+ BoT4− BoT8+ phenotype, were dependent on both specific antigen and T cell growth factor for proliferation and were restricted by determinants on class I major histocompatibility complex molecules. The clones killed target cells infected with either the Muguga or Marikebuni stocks of the parasite; the target cell lines tested included T cell clones which were infected in vitro with the two parasite stocks and subsequently recloned. The specificity of these cytotoxic T cell clones contrasts with that of T cell clones generated previously from animals immunized against T. parva (Muguga), in that the latter were specific for target cells infected with the Muguga stock of the parasite. Moreover, one of the clones generated against T. parva (Marikebuni) was restricted by the same major histocompatibility complex molecule as the Muguga‐specific T cell clones. The difference in parasite strain‐specificity between the two sets of clones appears to reflect the capacities of the two parasite stocks to cross‐protect, since animals immunized against T. parva (Marikebuni) are protected against challenge with T. parva (Muguga) whereas a proportion of animals immunized with T. parva (Muguga) are susceptible to challenge with T. parva (Marikebuni). Another difference between the two sets of T cell clones was that those generated against T. parva (Marikebuni) only killed a proportion of cells of a given cell line in a 4‐h cytotoxicity assay, whereas Muguga‐specific T cells invariably kill the majority of cells. However, despite this partial killing, the clones markedly inhibited growth of parasitized cell lines when cultured with them for a period of 5 days.
Cytotoxicity of tumor necrosis factor for thyroid epithelial cells and its regulation by interferon‐γ
Authors: Taverne Janice , Rayner David C. , van der Meide Peter H. , Lydyard Peter M. , Bidey Stephen P. , Cooke Anne 
Published: March 2007

AbstractThe FRTL‐5 line of differentiated rat thyroid epithelial cells was shown to be sensitive to the cytotoxic action of recombinant tumor necrosis factor. The sensitivity of the cells varied with the conditions of culture. It was markedly increased by preincubation with recombinant interferon‐γ, and the magnitude of this enhancement was affected by the presence of thyroid‐stimulating hormone. The increased susceptibility of the cells to the cytotoxicity of tumor necrosis factor was clear as early as 2–4 h after the addition of interferon‐γ and greatly preceded the induced expression of major histocompatibility complex class II antigen. Since cells secreting interferon‐γ and tumor necrosis factors may co‐exist in inflammatory infiltrates, our observations suggest that these factors may be early mediators of cell destruction in autoimmune thyroid disease.
Immunogenic presentation of an immunoglobulin “tolerogen” requires interleukin 2
Authors: Sadegh‐Nasseri Scheherazade , Phipps Richard P. 
Published: March 2007

AbstractPreviously it has been reported that hapten‐specific B cell unresponsiveness could be elicited by macrophages pulsed with the tolerogen deaggregated fluoresceinated sheep gamma globulin (FL‐SGG; R. P. Phipps and D. W. Scott, J. Immunol. 1983. 131: 2122). In contrast, FL‐SGG‐pulsed P388AD.2, a lymphoid dendritic cell‐like tumor line, presents this signal as an immunogenic one leading to augmented anti‐FL antibody responses. In the present study, we examined the role of T cells and their secreted lymphokines in the immunogenic presentation of FL‐SGG by P388AD.2. Lymphocytes and FL‐SGG‐pulsed P388AD.2 form large clusters in vitro. In order to examine whether the clustered lymphocytes were responsible for the augmented antibody responses, cultures of P388AD.2 and lymphocytes were separated into P388AD.2 adherent (clustered) and nonadherent fractions. Interestingly, the clustered fraction was entirely responsible for the augmented responses and contained Ly‐1+ T cells and B cells primed by FL‐SGG on P388AD.2. Moreover, a requirement for T cells in the presentation of a normally tolerogenic signal as an immunogenic one was demonstrated as responses of T‐depleted spleen cells could be reconstituted by addition of normal T cells or by an autoreactive T cell clone. Furthermore, the requirement for T cells could be bypassed using supernatant from concanavalin A‐stimulated spleen cells or by culture supernatant from AOFS, an ILZsecreting Tcell hybridoma. This suggested that a cognate interaction between T and B cells was not required to induce B cell priming and the augmented anti‐FL antibody responses. Further studies revealed that doses of recombinant IL2 > 12.5 units/ml, in conjunction with FL‐SGG‐pulsed P388AD.2, replaced the need for T cells. Overall, our data suggest that one mechanism of presentation of FL‐SGG in an immunogenic fashion involves T cell secretion of IL2 by autoreactive T cells triggered by close association with P388AD.2.
CDw40 and BLCa‐specific monoclonal antibodies detect two distinct molecules which transmit progression signals to human B lymphocytes
Authors: Clark Edward A. , Yip Timothy C. , Ledbettep Jeffrey A. , Yukawa H. , Kikutani Hitoshi , Kishimoto Tadamitsu , Ng Mun H. 
Published: March 2007

AbstractTwo monoclonal antibodies (mAb), MA6 and G28‐5, have the common property of detecting markers expressed on both B lymphocytes and carcinomas: BLCa (B lymphocyte carcinoma cross‐reacting antigen) and CDw40 (Bp50). A comparison of the reactivity of these mAb revealed that MA6 and G28‐5 detect distinct epitopes with different cell line and tissue distributions. L cell transfectants expressing CDw40 were not bound by MA6 anti‐BLCa, but were bound by G28‐5 anti‐CDw40. G28‐5 or a CDw40‐specific heterantiserum could not block the migration of BLCa, while MA6 antibody could. These results indicate that CDw40 and BLCa are distinct surface molecules. Both G28‐5 anti‐CDw40 and MA6 anti‐BLCa mAb could provide progression signals for B cells activated by appropriate B cell activators such as phorbol esters or anti‐immunoglobulin; however, only G28‐5 anti‐CDw4O and not MA6 was co‐stimulatory with the anti‐CD20 competence signal, demonstrating a clear difference in the CDw40 and BLCa‐mediated progression signals. Apparently, these molecules, although structurally distinct, have related functions in B cell activation.
Involvement of soluble mediator(s) different from interleukin (IL)1 in the antigen‐induced IL 2 receptor expression and proliferation of L3T4+ (CD4+) T lymphocytes
Authors: Germann Tieno , Schmitt Edgar , Huhn Helga , Rüde Erwin 
Published: March 2007

AbstractProliferation of T lymphocytes (T cells) requires the interaction of interleukin 2 (IL 2) with the high affinity form of the IL2 receptor (IL2R). IL2 production as well as IL 2R expression are generally induced simultaneously in T cells by the recognition of specific antigen displayed on the surface of syngeneic antigen‐presenting cells. The experiments described herein show that the expression of IL2R has different requirements than the production of IL2 (and other lymphokines). Stimulation of antigen‐specific L3T4+ T cell lines with antigen‐pulsed spleen cells (SC) treated with ultraviolet (UV) light results in efficient IL 2 production but only minimal proliferation due to reduced IL2R expression, as compared to T cells stimulated by antigen and SC without UV light treatment. The reduced IL 2R expression/proliferation correlates with the absence of a soluble mediator(s) termed T cell‐stimulating factor (TSF) in the supernatants of T cells stimulated with antigen‐pulsed, UV light‐irradiated SC. Addition of TSF of these T cells could at least partially restore the proliferative response or enhance IL2R expression. Because TSF is present in the supernatant of T cells triggered by antigen and SC but is absent when the latter are UV light treated, we suggest that TSF is a product of metabolically active splenic antigen‐presenting cells. The macrophage products interleukin 1α β p as well as some other cytokines show no TSF activity.
T cell proliferation induced by anti‐cd3 antibodies: requirement for a t‐t cell interaction
Authors: Bentin Jacques , Vaughan John H. , Tsoukas Constantine D. 
Published: March 2007

AbstractIn the presence of interleukin 2 (IL2), soluble anti‐CD3 monoclonal antibodies can stimulate highly purified normal T lymphocytes to proliferate. In these experiments HLADR+ T cells constituted 13 to 20% of the total cell population, and other HLADR− cells, such as monocytes and B lymphocytes, constituted less than 1% of the population. When the HLADR+ T cells were removed from the total T cell population by cytofluorometric sorting, the residual HLADR− T cells failed to respond to soluble anti‐CD3 plus IL2. When the separated HLADR+ T cells were recombined with the HLADR− T cells, a response was again found. This response was dependent on the dose of HLADR− T cells added in the mixture. Irradiation individually of the HLA DR+ and DR− T cells revealed that the proliferation in the cell mixture was predominantly, if not exclusively, by the HLADR− T cells. The ability of the HLA DR+ T cells to provide a signal necessary to this proliferation was radioresistant. These data indicate that under the conditions of these experiments HLADR+ (“activated”) T cells provide a signal necessary to the responsiveness of previously resting T cells.
The transmembrane orientation of the ϵ chain of the TcR/CD3 complex
Authors: Clevers Hans , Dunlap Sabrina , Terhorst Cox 
Published: March 2007

AbstractThe antigen receptor of the T lymphocytes is one of the most complex eukaryotic membrane structures studied to date. The T cell receptor (TcR) consists of two disulfide‐linked glycoprotein chains (α/βor γ/δ) and is noncovalently associated with a group of small and invariable CD 3 proteins. Four CD 3 chains have been recognized: two highly homologous glycoproteins CD 3 γ and δ, the more distantly related nonglycosylated CD 3 E chain, and the nonglycosylated CD 3 ζ, the latter being present as a homodimer. The unraveling of the architecture of the TcWCD 3 complex is crucial to our understanding of the processes underlying its assembly, recognition and trans‐membrane signaling. The transmembrane orientation of the TcR chains and of CD 3 γ and CD 3 δ can be directly inferred from their primary structure, based on the presence of concensus N‐linked glycosylation sites N‐terminal of their transmembrane domains. This prediction can not be made, however, for nonglycosylated molecules like the CD 3 E chain. In order to determine the transmembrane orientation of CD 3 E, anti‐peptide antisera directed against the N‐termini of the human and murine CD 3 E chains were generated in rabbits. Both antisera stained intact T cells, demonstrating that the N‐terminus of the CD 3 E chain was located at the outer surface of the plasma membrane. The anti‐human CD 3 E peptide antiserum was found to be mitogenic for peripheral blood T cells, a finding previously reported only for monoclonal anti‐TcW CD 3 reagents. Using a novel transient expression system in murine T lymphocytes, the human CD 3 E chain could be expressed on the surface of CD 3+, but not CD 3−murine T cells, as indicated by fluorescence staining with the anti‐peptide antiserum. This experiment confirmed the specificity of the anti‐peptide antiserum and, perhaps more importantly, indicated that the human CD 3 E chain was correctly assembled in the murine CD 3 complex. Moreover, the anti‐human CD 3 monoclonal antibody UCHT1 was found to stain T cells expressing the human CD 3 E chain.
Determination of frequency of T cells expressing the T cell‐specific serine proteinase 1 (TSP‐1) reveals two types of L3T4+ T lymphocytes
Authors: Fruth Uli , Nerz Gaby , Prester Marlot , Simon Hans G. , Kramer Michael D. , Simon Markus M. 
Published: March 2007

AbstractTSP‐1 is a murine T cell‐specific serine proteinase which is exclusively expressed in activated but not in resting T lymphocytes. Among T lymphocyte clones tested so far the enzyme was found to be associated with all Ly‐2+ but only with a fraction of L3T4+ lines. Here we have applied a limiting dilution system to determine the frequency of precursor cells of resting L3T4+ and Ly‐2+ lymphocytes which can be induced in vitro by antigen/lectin to express TSP‐1. T cell subsets were either positively enriched by flow cytofluorometry cell sorting or by negative selection using monoclonal antibodies and complement. Following stimulation of lymphocytes in vitro, individual microwells were tested for growth by visual examination and for the TSP‐1 protein/enzyme by analyzing cell lysates using either a specific rabbit anti‐TSP‐1 antiserum and/or the chromogenic model peptide substrate H‐D‐Pro‐Phe‐Arg‐p‐nitroanilide. Moreover, a large panel of L3T4+ and Ly‐2+ T lymphocyte clones generated from primary cultures were similarly investigated. In some cell cultures the presence of TSP‐1 was also tested on the mRNA level using a TSP‐1‐specific oligonucleotide probe. The data show that the majority, if not all, of antigenflectin‐induced‐Ly‐2+ T cells expressed TSP‐1. In contrast, only 12%‐27% of the growing lectin or antigen‐reactive L3T4+ lymphocytes were positive for the enzyme. Studies performed in parallel with L3T4+ and Ly‐2+ lymphocyte populations sensitized in bulk culture showed that under these conditions similar levels of TSP‐1‐specific mRNA and enzyme activity are detected in both subsets. The finding of primary L3T4+ T lymphocyte clones with distinct patterns of TSP‐1 production provides evidence for the existence of two types of L3T4+ effector cells with different functional capacities. The data also suggest a cooperation between distinct L3T4+ lymphocytes for induction of optimal TSP‐1 activity in L3T4+ T cells.
Recombinant human interferon‐γ induces increased IgE receptor expression on human platelets
Authors: Pancré VéRonique , Joseph Michel , Capron André , Wietzerbin Juana , Kusnierz Jean‐Pierre , Vorng Han , Auriault Claude 
Published: March 2007

AbstractHuman recombinant interferon‐gamma (IFN‐γ) significantly increased the expression of receptors for IgE (FceRII) on blood platelets. FceRII was measured by specific binding of 25I‐labeled IgE or flow cytometry experiments. Scatchard analysis of 125I‐labeled IgE binding curves revealed that treatment with IFN‐y increased the number of FceRII but did not change the value of the association constant of FCERII for l25I‐labeled IgE. IFN‐a had no effect on the expression or affinity of FcERII. In addition to FceRII, IFN‐γ also modified the expression of the glycoprotein IIb‐IIIa complex on the platelet membrane.
V genes of the primary antibody response of C57BL110 mice to the hapten phenyloxazolone
Authors: Kaartinen Matti , Pelkonen Eila , Even Jos , Mäkelä Oiii 
Published: March 2007

AbstractThe mRNA of ten monoclonal phenyloxazolone (phOx) antibodies originating from the primary (day 7) response of C57BL/10 mice were partially sequenced. The sequences were analyzed together with those of two previously published antibodies. The C57BL response does not have a predominant subset of antibodies like the BALB/c response has (VH‐OX1/VX‐OX1 JK5). Probably, C57BL mice lack the VH‐Ox1 gene and, as a consequence, their Vx‐Ox1 gene does not have a main role in the anti‐phOx response. Five Vx, and six VH genes were found to participate. All five Vx genes or their “alleles” had previously been found from the BALB/c response to 2‐phenyl‐oxazolone (phOx). On the other hand, the two strains use different VH genes for the anti‐phOx response. Most C57BL antibodies were coded by VH genes of group 1 which has only minor role in the BALB/c response. The remaining VH genes were from group 7. Our data show that one Vx, segment (e.g. Vx‐Ox1) can code for anti‐phOx antibodies with several, even widely different, VH genes. On the other hand, they emphasize the role of certain VH/VL gene combinations for the anti‐phOx specificity. Thus, VH genes of group 7 were found to code for anti‐phOx antibodies only together with the Vx45.1 gene.