bluuebunny/crossref_metadata_2025_split huggingface dataset:
Kinetic analysis of Ly‐6 gene induction in a T lymphoma by interferons and interleukin 1, and demonstration of Ly‐6 inducibility in diverse cell types
Authors: Leclair Kenneth P. , Bridgett Margot M. , Dumont Francis J. , Palfree Roger G. E. , Hämmerling Ulrich , Bothwell Alfred L. M. 
Published: March 2007

AbstractThe Ly‐6 locus contains multiple genes encoding cell surface proteins, two of which, when cross‐linked by antibodies, effect antigen‐independent activation of T lymphocytes. In this study, cDNA for Ly‐6‐encoded antigens have been used as probes to examine RNA from various tissues and transformed cell lines for constitutive levels of Ly‐6 RNA expression. Analyses of RNA prepared from several different tissues revealed a high level of expression of Ly‐6 RNA in kidney, spleen, heart and thymus, with a more moderate level of expression in liver, brain and lung tissue cells. A survey of various cell lines demonstrated the presence of Ly‐6 RNA in many, but not all T lymphocytic cell lines, in L cells, the Meth A fibrosarcoma, in the TCMK kidney cell line, and in the Neuro‐2a neuroblastoma. We also evaluated the expression of Ly‐6 RNA in cells after treatments with interferons (IFN) and interleukin 1 (IL 1). Treatment of lymphoid cells with IFN (α/β and γ), known to increase cell surface Ly‐6 antigen expression in normal T cells, was correlated with increases in Ly‐6 RNA levels. Increases in levels of RNA correlated with increases in levels of the Ly‐6A/E or Ly‐6C antigens. Several T lymphoid cell lines exhibiting Ly‐6 RNA inducibility by IFN were similarly inducible with IL 1. Kinetic experiments using one such line, (YAC‐1), showed that the induction of Ly‐6 RNA mediated by IFN‐α/β occurred rapidly (within 4 h), while the induction by IL 1 required relatively more time (∼︁ 8 h). Although the actions of IFN‐α/β were not blocked by cycloheximide, the presence of this protein synthesis inhibitor significantly attenuated the effects of IL 1 and IFN‐γ on Ly‐6 RNA transcription. Induction by IFN‐γ as well as IL 1 could be blocked completely by co‐culture with anti‐IFN‐γ, implicating IFN‐γ as a mediator of the induction by IL 1.
Lymph node macrophage heterogeneity: the phenotypic and functional characterization of two distinct populations of macrophages from rat lymph node
Authors: Chao David , MacPherson G. Gordon 
Published: March 2007

AbstractThe isolation and characterization of lymph node macrophages (MΦ) has shown a hitherto unknown heterogeneity. Two types of MΦ were distinguished by morphology, monoclonal antibody staining and functional assays. The type I MΦ failed to express surface Ia even when activated, a characteristic which has only previously been reported for splenic marginal zone MΦ despite studies suggesting an antigen presentation role for the MΦ, the failure to express surface Ia would seem to eliminate an interaction with T helper cells for the type I MΦ in the lymph node. In contrast, the type I MΦ, other characteristics of clustering with activated B cells in vitro, the colocalization of the type I MΦ and activated B cells in situ, the specific uptake of thymus‐independent type 2 antigens and the failure to undergo respiratory burst activity all suggest a MΦ‐B cell interaction, possibly of a trophic nature.The defective microbicidal activity of the type I MΦ may have been compensated for by the type II MΦ, which expresses both strong respiratory burst activity and surface Ia expression when freshly isolated. However, unlike the inflammatory MΦ the activated phenotype of the type II MΦ did not appear to be interferon‐γ dependent because type II MΦ could also be isolated from nude rat lymph nodes.
Differential pattern of T cell recognition of the 65‐kDa mycobacterial antigen following immunization with the whole protein or peptides
Authors: Brett Sara J. , Lamb Jonathan R. , Cox Josephine H. , Rothbard Jonathan B. , Mehlert Angela , Ivanyi Juraj 
Published: March 2007

AbstractThe 65‐kDa stress protein from Mycobacterium bovis (Bacillus Calmette Guérin) elicited T cell proliferation and antibody responses in seven B10 congenic mouse strains with different H‐2 haplotypes. To analyze T cell determinants on this antigen, seven peptides corresponding to six predicted T cell epitopes, and one defined B cell epitope were synthesized. Mice were either immunized with the whole antigen and the specificity of the response was ascertained in respect of the six peptides, or mice were immunized with seven of the peptides and tested for proliferative responses to the whole molecule. The results showed that three peptides carried epitopes to which mice responded following injection of the whole molecule and that immunization with two additional peptides could prime for in vitro stimulation with the native antigen. The latter result indicates the feasibility of generating T cell responses to “cryptic” epitopes on proteins by immunizing with peptides. The peptide‐specific T cell responses were distinctly influenced by the H‐2 haplotype of mouse strains. However, two peptides were recognized by several H‐2‐disparate mouse strains, and one peptide could be presented by both I‐A and I‐E molecules. Immunization with several peptides induced a cross‐reactive T cell proliferative response to the homologous GroEL protein isolated from E. coli. The amount of cross‐reactivity was influenced by the extent of sequence homology between mycobacterial and E. coli proteins and the major histocompatibility complex class II molecule used to present the peptide.
Suppressive effect of interferon‐γ on the BCL1 cell‐dependent interleukin 5 bioassay
Authors: Lohoff Michael , Sommer Frank , Röllinghoff Martin 
Published: March 2007

AbstractProliferation of BCL1 cells, generally used for the measurement of murine interleukin (IL)5, is shown to be strongly suppressed by interferon (IFN)‐γ. This effect can be abrogated by the addition of anti‐IFN‐γ antibodies. Thus, the IL5 activity of culture supernatants derived from T cells simultaneously producing IFN‐γ and IL 5, can only be seen with BCL1 cells in the presence of anti‐IFN‐γ antibodies. If anti‐IFN‐γ antibodies are omitted, T cells which produce both, IFN‐γ and IL5, may mistakenly be grouped into the TH1 subtype producing only IFN‐γ, but no IL5.
Expression of disulfide‐linked and non‐disulfide‐linked forms of the T cell receptor γ/δ heterodimer in human intestinal intraepithelial lymphocytes
Authors: Spencer Jo , Isaacson Peter G. , Diss Tim C. , Macdonald Thomas T. 
Published: March 2007

AbstractImmunohistochemistry has been used to investigate disulfide‐ and non‐disulfide‐linked forms of the T cell receptor γ/δ heterodimer (TcR γ/δ) in blood and intestinal epithelium of normal human small intestine, intestine of patients with untreated coeliac disease (in whom T cells expressing TcR γ/δ are disproportionately raised), intestine of patients with tropical malabsorption, and in the human fetus. In blood from adult volunteers, 90% of T cells expressing TcR γ/δ use the disulfide‐linked form. In contrast in the epithelium in normal small intestine, coeliac disease and tropical malabsorption, most of the T cells expressing TcR γ/δ use the non‐disulfide‐linked form. This is especially prominent in untreated coeliac disease where the increase in TcR γ/δ T cells is mainly restricted to those using the non‐disulfide‐linked form. In human fetal small intestinal epithelium, however, only cells using the disulfide‐linked form are present. These variations in expression of different forms of TcR γ/δ in the gut epithelium in different conditions suggests that antigen, or some as yet undefined factor may determine the frequency of each subpopulation.
Decline in growth and antibody production of established hybridomas and transfectomas with addition of interleukin 6
Authors: Karavodin Linda M. , Glasebrook Andrew L. , Phelps Julia L. 
Published: March 2007

AbstractSeveral sources of the lymphokine interleukin (IL) 6 (human recombinant IL6, supernatant from P388D1, a murine macrophage cell line and murine thymocyte‐conditioned media) were examined for enhancement of growth, immunoglobulin (Ig) secretion and seeding density requirements in murine hybridoma and transfectoma cell lines established in the absence of exogenously added IL 6. None of these preparations increased growth or Ig secretion in the six cell lines studied. In fact, inhibition of proliferation was seen with two of the three IL 6 preparations. Human recombinant IL6 showed no effect on minimal seeding density, but murine thymocyte‐conditioned media gave a tenfold decrease in minimal seeding density requirements in one of two hybridomas and one of two transfectomas studied. All preparations contained active IL6, as measured by proliferation of the IL6‐dependent cell line T1165.D10. These data show that IL6 alone does not increase growth or Ig secretion for these established cell lines. This finding is compared with previous reports describing increased growth in primary cultures of murine hybridomas and human myelomas with IL6.
A cytokine, lymphocyte blastogenesis inhibitory factor (LBIF), arrests mitogen‐stimulated T lymphocytes at early G1 phase with no influence on interleukin 2 production and interleukin 2 receptor light chain expression
Authors: Sugimura Kazuhisa , Ueda Yutaka , Takeda Koji , Fukuda Shigeharu , Tsukahara Kappei , Habu Yoshiki , Fujiwara Hiroshi , Azuma Ichiro 
Published: March 2007

AbstractThe function of a human cytokine, lymphocyte blastogenesis inhibitory factor (LBIF), was characterized. To this end, LBIF was purified from crude supernatant of U‐937 cells, a human macrophage‐like cell line, by using fast protein liquid chromatography (FPLC).We demonstrated here that (a) the LBIF preparation completely inhibited phytohemagglutinin (PHA)‐stimulated T cell proliferation; (b) however, in PHA‐stimulated T lymphocytes LBIF inhibited neither interleukin (IL)2 production nor the expression of IL2 receptor (IL2R) light chain (CD25) which play a critical role for T cell proliferation; (c) LBIF arrested PHA‐stimulated T lymphocytes at the G1 phase of cell cycle and inhibited entry into S phase, thus inhibiting lymphocyte proliferation. Wright‐Giemsa's staining of the cells showed that PHA/LBIF‐stimulated cells were arrested in early G1. In agreement with this result, LBIF strongly inhibited PHA‐induced RNA synthesis. Further, LBIF inhibited the induction of the transferrin receptor which is normally expressed at the late G1 phase of the cell cycle. The inhibitory activity of LBIF was reversible. Thus, this study elucidated that LBIF arrests PHA‐stimulated T lymphocytes at a point between the stages of IL 2 production or IL2R light chain expression (in early G1) and transferrin receptor expression (in late G1). Taken together, these results suggest that there might be a control system of T cell proliferation distinct from the previously reported mechanisms, such as the inhibition of IL2 production or the inhibition of IL 2R light chain (CD25) expression, and that LBIF might be an important molecule in the regulation of normal lymphocyte proliferation.
The biochemical characterization of E2, a T cell surface molecule involved in rosettes
Authors: Aubrit Françoise , Gelin Catherine , Pham Danièle , Raynal Brigitte , Bernard Alain 
Published: March 2007

AbstractWe have previously identified a molecule on the T cell surface, which, in addition to CD2 is involved in the rosette phenomenon. This is a 32‐kDa single polypeptide chain which we have termed E2. The studies reported here show striking patterns on the glycosylation status of E2. It is a heavily sialylated and glycosylated molecule, the sugar moieties accounting for almost half of its relative molecular mass (Mr). It carries no N‐linked sugar residues, only O‐linked oligosaccharides. Despite heavy glycosylation, the molecule appears to behave homogeneously on gel electrophoresis, both in terms of Mr and pI. Neuraminidase treatment of E2 lowered its Mr to 28 kDa; this was further decreased to 18 kDa after removal of O‐linked sugar residues by treatment with O‐glycanase. An identical reduction in size was observed after treatment with trifluoromethane sulfonic acid, showing that the molecule carries no detectable N‐linked sugar residues. Moreover, endoglycosidase F and endoglycosidase H treatment of either the immunoprecipitates from 125I surface‐labeled thymocytes, or of a purified preparation of E2, did not reduce the Mr of E2, nor did tunicamycin treatment of T cells. Two‐dimensional gel electrophoresis revealed two discrete spots of acidic pI (4.4 and 4.6) that were still seen after neuraminidase treatment, though they had moderately shifted. Pulse‐chase experiments revealed a single 28‐kDa precursor form that could have been the unsialylated molecule. Finally, sequencing 14 amino acid residues of the N‐terminal side revealed no homology with known proteins. Since the sugar moieties of adhesion protein could play an important role, the results obtained in this study will prove valuable to our understanding of the role exerted by the E2 molecule.
Bispecific F(ab′)2 monomer prepared with anti‐CD3 and anti‐tumor monoclonal antibodies is most potent in induction of cytolysis of human T cells
Authors: Nitta Taizo , Yagita Hideo , Azuma Takachika , Sato Kiyoshi , Okumura Ko 
Published: March 2007

AbstractInduction of cytolytic activity of peripheral blood mononuclear cells towards target cells was studied by preparing bispecific F(ab′)2 which was composed of two Fab′ fragments, one of which was derived from anti‐CD3 monoclonal antibody and the other from anti‐tumor monoclonal antibody. After reduction of the interchange disulfide bonds of these fragments by dithiothreitol, a thiol‐disulfide interchain reagent, 5,5′‐dithiobis‐2‐nitrobenzoic acid, was added to convert the free SH groups of one of the Fab′ fragments to mixed disulfide derivatives. These were then coupled with the other Fab′ fragment bearing free SH groups, producing a bispecific hybrid F(ab′)2 monomer of high yield. The bispecific F(ab′)2 monomers were able to render nonactivated peripheral blood mononuclear cells cytotoxic against natural killer‐resistant tumor cell lines at doses as low as 1 μg/ml. The polymeric forms of the F(ab′)2 fragments prepared by use of cross‐linking reagents such as N‐succinimidyl‐3‐(2‐pyridyldithiol)‐propionate (SPDP) or S‐acetylmercaptosuccinic acid anhydride (SAMSA) were less efficient for induction of cytolytic activity than the monomeric one. It may be feasible to use bispecific F(ab′)2 monomers in cancer immunotherapy because of the ease of preparation, as well as the efficiency in inducing cytolytic activity and their high tissue permeability due to their small size (100–110 kDa). In addition, redirected cytolytic activity towards peripheral blood mononuclear cells by reticuloendothelial system cells, resulting from linking these two types of cells through Fc‐Fc receptor interactions, does not occur.
Cytoplasmic CD3+ surface CD8+ lymphocytes develop as a thymus‐independent lineage in chick‐quail chimeras
Authors: Bucy R. Pat , Coltey Monique , Chen Chen Lo H. , Char David , Douarin Nicole M. Le , Cooper Max D. 
Published: March 2007

AbstractWe have analyzed the embryonic development of a population of lymphoid cells that express a CD3 antigenic determinant in the cytoplasm but not on the cell surface. Since these cells lack T cell receptor (TcR) molecules, we have provisionally named them TCRO cells. Their development, expansion and distribution was investigated following transplantation of splenic and bursal fragments from chicken embryos into quail embryos. Since quail cells are not recognized by our panel of monoclonal antibodies against chicken TcR1, TcR2, TcR3, CD3, CD4 and CD8 molecules, these antibodies provided reliable markers for donor chick lymphocytes in the tissues of the quail recipients. Transplanted spleen and bursa both generated CD3+ cells, the number of which increased as a function of age. Notably, approximately half of these CD3+ cells expressed surface CD8, but none acquired TcR1 (γ/δ), TcR2 (α/β) or TcR3 expression. Since TCRO cells normally appear first in the spleen of 8‐day chick embryos (E8), their generation in E6 splenic transplants indicated an extrathymic origin. The TCRO cells of chick splenic origin migrated to the spleen, bursa and thymus of the quail recipients. In six of seven chimeras acquiring CT3+ cells in the recipient thymus, these cells were restricted to the medulla and displayed the typical TCRO phenotype: CD3+CD8+TcR1‐TcR2‐TcR3‐. These intrathymic TCRO cells also lacked the CT1 thymocyte antigen. We conclude that the TCRO cells represent a thymus‐independent lineage of lymphoid cells that can migrate into a receptive thymus but rarely, if ever, differentiate into conventional T cells.
Fate and biological action of human recombinant interleukin 1β in the rat in vivo
Authors: Klapproth Jan , Castell José , Geiger Thomas , Andus Tilo , Heinrich Peter C. 
Published: March 2007

AbstractThe plasma half life of recombinant human interleukin 1β (rhIL 1β) was determined in rats by measuring the disappearance of the radioactivity of 125I‐labeled rhIL 1β from the circulation. The plasma clearance showed a biphasic behavior: an initial fast disappearance (half life of about 3 min) was followed by a second slower one (half life of about 4 h). Twenty minutes after a single‐dose injection of 125I‐labeled rhIL 1β most of the radioactivity was concentrated in kidneys, liver and intestine.rhIL 1β induced the synthesis of α1‐acid glycoprotein (AGP), α1‐cysteine proteinase inhibitor (CPI) and β‐fibrinogen mRNA in liver. Half maximal stimulation was elicited by approximately 3000 U of rhIL 1β per animal. The mRNA changes for AGP and CPI were followed by corresponding protein increases in serum. Twenty hours after rhIL 1β injection, serum AGP rose from 0.7 to 2.5 mg/ml. CPI increased from 0.3 to 1.9 mg/ml 25 h after administration of rhIL 1β. Within 20 h after rhIL 1β injection, albumin serum concentration showed a strong decrease, preceded by a reduction in hepatic albumin mRNA levels. Neither changes in albumin synthesis nor degradation can explain this decrease suggesting that other mechanisms such as increased transvascular permeability are involved.
Inhibition by chlorpromazine of lymphokine‐specific mRNA expression in human thymocytes
Authors: Schleuning Michael J. , Duggan Anne , Reem Gabrielle H. 
Published: March 2007

AbstractThis study was designed to determine the effect of the phenothiazine chlorpromazine (CPZ) on the activation of human thymocytes. We provide evidence that CPZ inhibits the accumulation of mRNA specific for the lymphokines interleukin 2, interferon‐γ, tumor necrosis factor α and the proto‐oncogene c‐myc; by contrast, the accumulation of mRNA specific for the α chain of the interleukin 2 receptor and the subsequent early expression of Tac antigen on the cell surface is not inhibited by CPZ. The inhibition of the expression of lymphokine‐specific mRNA results in a decrease in interferon‐γ synthesis and in inhibition of thymocyte proliferation as determined by the incorporation of [3H]thymidine. In addition, we show that activation of protein kinase C (PKC) in human thymocytes by 12‐O‐tetradecanoyl phorbol 13‐acetate (TPA) causes the phosphorylation of a protein of a molecular mass of approximately 75 kDa. The function of this protein is as yet not defined, but it is possible that it plays a role in the transduction of the signals to the nucleus which in turn elicit the expression of the genes coding for c‐myc and for the lymphokines required for thymocyte activation. We also demonstrate that CPZ, like the immunosuppressant drug cyclosporin A does not inhibit the phosphorylation of the 75‐kDa protein which is induced by the activation of PKC by TPA and does not affect phosphoinositide breakdown, indicating that it exerts its effect at a site distal to the activation of PKC. These observations demonstrate that CPZ has an immunoregulatory function in addition to its psychotropic activity.
Analysis of T cell receptor γ chains from adult CD4‐CD8‐ thymocytes
Authors: Moore Mark W. , Crispe I. Nicholas , Bevan Michael J. 
Published: March 2007

AbstractThe role of T cell receptor (TcR) γ genes in T cell development has not been determined. To extend our understanding of the repertoire of TcR γ expression, we prepared a cDNA library from CD4‐CD8‐ adult BALB/c thymocytes and cloned and sequenced 15 TcR γ genes from this cDNA library. We found that two clones were transcripts of the unrearranged Cγ2 gene and that three clones terminated in the Jγ2 region. Nine of the remaining clones were Vγ1.2Jγ2Cγ2 genes and five of these were in frame. Only one clone corresponded to Cγ1 and resulted from an in‐frame Vγ2Jγ1Cγ1 join. Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis analysis of the γ chain proteins from the surface of both BALB/c and C57BL/6 adult CD4‐CD8‐ thymocytes did not detect the 32‐kDa Vγ1.2Cγ2 protein, but did detect the 35‐kDa VγCγ1 protein. These results suggest that despite the abundance of full‐length functional Vγ1.2Cγ2 transcripts in the thymocyte subset, the protein product is not expressed on the cell surface as the predicted 32‐kDa γ protein. Finally, our analysis of the V‐J joining of the γ genes reveals both flexibility at the V‐J junction and extensive N‐region nucleotide addition that lead to diversity of the predicted protein sequence.
Species specificity of interleukin 2 binding to individual receptor components
Authors: Collins Mary K. L. 
Published: March 2007

AbstractThe affinity of recombinant human and murine interleukin 2 (IL 2), which differ significantly in structure in the amino terminal α helix, for the human or murine interleukin 2 receptors was compared. Murine IL 2 showed a 10‐fold higher affinity than human IL 2 for the murine high‐affinity receptor expressed on T lymphocytes, whereas human IL 2 bound 100‐fold more strongly than murine IL 2 to the human high‐affinity receptor. As the high‐affinity IL 2 receptor on T lymphocytes has been demonstrated to be composed of two IL 2 binding components, p55 and p75, the role of the individual chains in the determination of species specificity was then investigated. Recombinant retroviral vectors were used to generate populations of 3T3 fibroblasts expressing either human or murine p55 in isolation, and their binding characteristics were determined. While the murine p55 showed an identical species preference to the murine p55/p75 high‐affinity complex, the human p55 had an equal affinity for human and murine IL 2. This suggested that the human p75 chain was responsible for the preference for human IL 2 shown by the human high‐affinity receptor, which was confirmed by performing a binding experiment using gibbon MLA 144 cells, expressing only p75. These data suggest that the amino terminal region of the IL 2 molecule interacts with the p75 chain of the IL 2 receptor.
Tolerance induction in the organ‐cultured thymus lobes upon intimate contact with allogeneic thymus lobes
Authors: Suzuki Gen , Kawase Yoshiko , Hirokawa Katsuiku 
Published: March 2007

AbstractIn an organ‐cultured murine fetus thymus, precursor cytotoxic T lymphocytes (pCTL) specific for alloantigens developed successfully but those reactive with self antigens were eliminated. In attempting to dissect the mechanism of self tolerance, intrathymic chimera was made by culturing two genetically disparate thymuses in close contact with each other (parabiosis of thymuses). This maneuver resulted in the induction of specific and mutual CTL tolerance. It seems that CTL tolerance was induced by clonal deletion but not by active suppression. Since 2'‐deoxyguanosine treatment abolished the tolerogenic capability of the thymus, hemopoietic cells capable of migrating to and fro in the parabiotic thymuses are thought to be responsible for tolerance induction. Induction of CTL tolerance was dependent on the maturation stages of T cells in the thymus: T cells in 5‐day‐, but not in 7‐day‐cultured thymuses were susceptible to tolerance induction, indicating that T cells expressing T cell receptors at low density are susceptible to tolerance induction.