bluuebunny/crossref_metadata_2025_split huggingface dataset:
Interleukin‐4 and RANTES expression in maturing eosinophils derived from human cord blood CD34+ progenitors
Authors: Velazquez J. R. , Lacy P. , Mahmudi‐Azer S. , Bablitz B. , Milne C. D. , Denburg J. A. , Moqbel R. 
Published: March 2003

SummaryEosinophils elaborate a number of proinflammatory mediators, including immunoregulatory cytokines and chemokines. Interleukin (IL)‐4 and RANTES are important cytokines that have previously been shown to be expressed by mature eosinophils. We hypothesized that de novo synthesis of IL‐4 and RANTES occurs in nascent eosinophils, leading to storage of newly produced proteins in crystalloid granule‐like structures. Cytokine mRNA and protein expression were examined in cultured eosinophil colonies, which were derived from purified cord blood CD34+ cells and generated in semisolid media (methylcellulose) in the presence of recombinant human (rh)IL‐3 and rhIL‐5. Cytokine mRNA profiles were analysed by the reverse transcription–polymerase chain reaction (RT–PCR) to determine transcription of IL‐4 and RANTES in cells on days 0, 7, 14, 21 and 28 of culture. The expression of translated cytokine products and granule major basic protein (MBP) was confirmed, from day 23 onwards, for colonies cultured in semisolid media, by immunofluorescent labelling and confocal laser‐scanning microscopy (CLSM). We found that mRNA sequences encoding IL‐4 and RANTES were expressed in freshly prepared, non‐differentiated CD34+ cells. Furthermore, RANTES mRNA localized to carbol chromotrope 2R‐positive colony cells, as assessed using in situ RT–PCR on day 21 of culture in semisolid media, and was found to gradually decrease (relative to β2‐microglobulin) in rhIL‐3‐ and rhIL‐5‐treated colony cells (comprising > 90% eosinophil‐like cells) up to day 28. Immunoreactivity for IL‐4 and RANTES co‐localized with MBP in maturing colony eosinophils on day 23 of culture in semisolid media, as judged by CLSM. These results suggest that synthesis and storage of immunoregulatory cytokines, essential for processes associated with adaptive immunity, occurs in nascent eosinophils during their growth and differentiation.
Cellular interactions in bovine tuberculosis: release of active mycobacteria from infected macrophages by antigen‐stimulated T cells
Authors: Liébana E. , Aranaz A. , Aldwell F. E. , Mcnair J. , Neill S. D. , Smyth A. J. , Pollock J. M. 
Published: March 2003

SummaryThe outcome of Mycobacterium bovis infections depends on the interactions of infected macrophages with T lymphocytes. Several studies in humans and in mouse models have suggested an important role for cytotoxicity in the protective immune response to mycobacterial infections, and both CD4+ and CD8+ T cells have been shown to elicit appropriate cytolytic activity. The present study investigated in vitro interactions of T cells with M. bovis‐infected macrophages in bovine tuberculosis. The results showed that following interaction with antigen‐stimulated peripheral blood mononuclear cells (PBMC) from infected cattle, there was an increased presence of M. bovis in the extracellular compartment of infected macrophage cultures, as measured by incorporation of [3H]uracil into mycobacterial RNA. Furthermore, out of a panel of T‐cell clones from infected cattle, it was found that a higher proportion of CD8+ clones produced an increase in the number of metabolically active extracellular M. bovis organisms compared with CD4+ clones. Finally, a positive correlation between percentage of antigen‐dependent release of mycobacteria and total uracil uptake by M. bovis within culture systems was detected. This could be regarded as an indication of preferential intracellular control of mycobacteria by activated macrophages.
Activation of bovine B cells via surface immunoglobulin M cross‐linking or CD40 ligation results in different B‐cell phenotypes
Authors: Haas K. M. , Estes D. M. 
Published: March 2003

SummaryExperiments reported herein demonstrate that activation of bovine B cells via surface immunoglobulin M (sIgM) cross‐linking, analogous to T‐cell independent (TI‐2) antigenic stimulation, results in the expression of CD5. Interestingly, in the presence of CD40 ligand, sIgM‐mediated induction of CD5 on B cells was inhibited. These findings indicate that activation of bovine B cells via B‐cell receptor (BCR) cross‐linking results in a CD5+B‐cell phenotype and that CD40 signalling is inhibitory to this process. Analysis of cytokine mRNA indicates that bovine B cells constitutively express tumour necrosis factor‐α (TNF‐α) and interleukin (IL)‐1β transcriptsin vitro, while IL‐10 mRNA expression is induced following sIgM cross‐linking. IL‐12 p40 transcripts were produced by B cells activated by CD40, but not by BCR, ligation. Analysis of cytokine receptor mRNA indicates that activation through CD40, in the presence or absence of IgM cross‐linking, results in increased IL‐4 receptor‐α (IL‐4Rα), IL‐13Rα1 and interferon‐α receptor 1 (IFN‐αR1) mRNA levels. Overall, these findings suggest that activation of bovine B cells through BCR cross‐linking yields an activation phenotype that differs substantially from that of B cells activated through CD40.
Monoclonal anti‐double‐stranded DNA autoantibody stimulates the expression and release of IL‐1β, IL‐6, IL‐8, IL‐10 and TNF‐α from normal human mononuclear cells involving in the lupus pathogenesis
Authors: Sun K.‐H. , Yu C.‐L. , Tang S.‐J. , Sun G.‐H. 
Published: March 2003

SummaryIn our previous reports, we found polyclonal anti‐double‐stranded DNA antibodies (anti‐dsDNA) purified from patients with active systemic lupus erythematosus (SLE) exerted inhibitory effect on [3H]thymidine incorporation of human mononuclear cells (MNC). However, the other immunological effects of anti‐dsDNA on the functions of MNC have not yet been reported. In this study, two monoclonal antibodies, 12B3 and 9D7, with different anti‐dsDNA activity were evaluated for their effects on the expression and release of different cytokines from human MNC. We confirmed absence of endotoxin in the two monoclonal antibody preparations and the used medium as detected by Limulus amoebocyte lysate test. The mRNA expression and release of different cytokines including interleukin (IL)‐1β, IL‐2, IL‐4, IL‐6, IL‐8, IL‐10, tumour necrosis factor‐α (TNF‐α) and interferon‐γ (IFN‐γ) were measured. We found the two monoclonal anti‐dsDNA not only dose‐responsively suppressed the phytohaemagglutinin (PHA)‐induced thymidine uptake of human MNC but stimulated the mRNA expression of IL‐1β, IL‐6 and IL‐8 in normal human MNC detected by reverse transcription–polymerase chain reaction (RT–PCR). Enzyme‐linked immunosorbent assay (ELISA) measurement of cytokines in MNC culture supernatants revealed that anti‐dsDNA enhanced IL‐1β, IL‐8, TNF‐α and IL‐10 release from resting MNC. These effects of anti‐dsDNA antibodies were not affected by polymyxin B, a potent binder and neutralizer of lipopolysaccharide (LPS). These in vitro studies suggest that anti‐dsDNA possess a dual effect on normal human MNC: (a) to enhance the release of proinflammatory cytokines (IL‐1β, IL‐8 and TNF‐α) from MNC to augment inflammatory reaction; and (b) to polarize the immune reaction towards the T helper 2 (Th2) (increased IL‐10 production) pathway. This unique effect of anti‐dsDNA may play a role in lupus pathogenesis by augmenting inflammatory reactions and autoantibody production which are commonly found in patients with active SLE.
Evidence for multiple CD95–CD95 ligand interactions in anteriorchamber‐associated immune deviation induced by soluble protein antigen
Authors: Kezuka T. , Streilein J. W. 
Published: March 2003

SummaryWe have investigated whether CD95–CD95 ligand interactions are important in anterior chamber‐associated immune deviation (ACAID) induced by soluble protein antigen, and if so, to identify the participating cells on which these molecules are expressed. Peritoneal exudate cells as antigen‐presenting cells (APC) obtained from B6.lpr/lpr, B6.gld/gld and C57BL/6 mice were cultured with ovalbumin (OVA) and transforming growth factor‐β2 (TGF‐β2) overnight, then injected intravenously into C57BL/6 or B6.lpr/lpr recipients. Some B6.lpr/lpr mice were reconstituted with naive T cells from wild‐type C57BL/6 donors. In other experiments, B6.lpr/lpr and B6.gld/gld mice received an anterior chamber injection of OVA followed 7 days later by subcutaneous immunization with OVA plus adjuvant. Delayed hypersensitivity (DH) was assessed with an ear swelling assay. T cells activated in vitro with OVA‐pulsed, TGF‐β‐treated APC were tested in vivo for their capacity to suppress DH expression in a local adoptive transfer assay. The results indicate that when ACAID was induced by in‐vitro generated ACAID‐inducing cells, the APC expressed CD95L, and recipient T cells expressed CD95. The capacity of in vitro generated regulatory T cells to suppress DH expression to OVA in vivo was not governed by CD95–CD95L interactions. When OVA was injected into the anterior chamber of naive mice, CD95 expression was required for ACAID induction, although ACAID was readily induced in CD95L‐deficient mice. We conclude that CD95–CD95L interactions are required in ACAID for the initial stage of APC presentation of eye‐derived antigens to T cells, and that CD95–CD95L interactions participate at one or more additional step in the process by which ACAID is induced by soluble protein antigens.
Analysis of histamine‐producing cells at the late phase of allergic inflammationin rats
Authors: Shiraishi M. , Hirasawa N. , Oikawa S. , Kobayashi Y. , Ohuchi K. 
Published: March 2003

SummaryTo identify histamine‐producing cells at the late phase of allergic inflammation, the expression of l‐histidine decarboxylase (HDC) was examined in the infiltrating leucocytes in the inflammatory locus. HDC activity and HDC mRNA levels in the infiltrating leucocytes in the pouch fluid of the immunized rats (that were injected with the antigen solution into the air pouch) were increased compared with those in the infiltrating leucocytes of the non‐immunized rats. When infiltrating leucocytes collected 8 hr after antigen injection were cultured, histamine production by the cells from the immunized rats was higher than that from the non‐immunized rats. In situ hybridization of HDC mRNA revealed that almost all the infiltrating leucocytes of the immunized rats, 4 hr after injection of the antigen, expressed HDC mRNA with high intensity, while those of the non‐immunized rats showed only a weak intensity of HDC mRNA. In the immunized rats, ≈ 90% of leucocytes infiltrating in the pouch fluid at 4 hr were neutrophils and 8% were monocytes/macrophages. Neither mast cells nor basophils were detected in the infiltrating leucocytes. When rat peritoneal neutrophils were incubated in the presence of 12‐O‐tetradecanoylphorbol 13‐acetate, histamine production was significantly increased. These findings suggest that the leucocytes, mainly neutrophils, infiltrating at the inflammatory locus are responsible for histamine production at the late phase of allergic inflammation.
Adjuvant effects of CpG oligodeoxynucleotides on responses against T‐independent type 2 antigens
Authors: Kovarik J. , Bozzotti P. , Tougne C. , Davis H. L. , Lambert P.‐H. , Krieg A. M. , Siegrist C.‐A. 
Published: March 2003

SummaryOligodeoxynucleotides containing CpG motifs (CpG‐ODN) are potent in vitro B‐cell activators and they have been successfully used to increase in vivo antibody responses to T‐dependent peptide and protein antigens. In contrast, the use of CpG‐ODN to enhance in vivo antibody responses to various T‐independent type 2 (TI‐2) antigens has recently generated contradictory results. In this study, we compared the CpG‐ODN stimulatory effect on antibody responses of adult and young BALB/c mice to trinitrophenylaminoethyl‐carboxymethyl (TNP) ‐Ficoll and to polysaccharides (PS) from several distinct serotypes of Streptococcus pneumoniae (SPn). CpG‐ODN co‐administration significantly enhanced antigen‐specific immunoglobulin M (IgM), IgG, IgG1 and IgG2a titres to TNP‐Ficoll. The depletion of CD4+ cells by monclonal antibodies (GK1.5) identified their essential role in CpG‐ODN‐mediated enhancement of antibody responses. In contrast to TNP‐Ficoll, CpG‐ODN failed to enhance IgM and IgG responses to any of the 18 SPnPS serotypes tested. Providing T‐cell epitopes by the conjugation of SPnPS to the carrier protein tetanus toxoid again allowed CpG‐ODN to mediate enhancement of IgG, IgG2a and IgG3 responses to most SPnPS serotypes. Thus, antigen‐presenting cell/T‐cell interaction appears to largely mediate the in vivo influence of CpG‐ODN on antibody responses to TI‐2 antigens. In early life, additional factors limit CpG‐ODN modulation of antibody responses to TI‐2 antigens.
The immune dysregulatory compound mercuric chloride induces integrin‐mediated T‐lymphocyte adhesion
Authors: Roos A. , Neeft M. , Engelen L. , Schilder‐Tol E. J. M. , Kunzendorf U. , Weening J. J. , Aten J. 
Published: March 2003

SummaryExposure of Brown Norway rats to mercuric chloride induces systemic autoimmunity, involving T‐ and B‐lymphocyte activation, (auto‐)antibody production and multiorgan inflammation. Several divalent metal ions, such as Mg2+ and Mn2+, can activate binding of integrins to their ligands, thus causing lymphocyte adhesion. To test the hypothesis that Hg2+ acts in a similar way, we studied the effect of HgCl2 on integrin‐mediated T‐cell adhesion. HgCl2 induced cell–cell aggregation of human T lymphoblasts. Exposure of a human T‐cell clone to HgCl2 for 1 hr enhanced, in a dose‐dependent way, cell binding to fibronectin (FN) and to intercellular adhesion molecules (ICAM) ‐1, ‐2 and ‐3. Furthermore, HgCl2 induced strong binding of Jurkat T cells to FN. These effects of HgCl2 were of similar magnitude as the effects of phorbol 12‐myristate 13‐acetate (PMA) or MnCl2. Studies using blocking antibodies indicated the involvement of CD11a in binding to ICAMs, and of CD49d, CD49e, and CD29 in binding to FN. Adhesion to FN induced by HgCl2 or by PMA, but not by MnCl2, was dependent on temperature and on extracellular Ca2+ or Mg2+. Addition of cytochalasin B enhanced synergistically the FN adhesion induced by MnCl2, whereas the effects of PMA and HgCl2 were not modified. These results indicate that Hg2+ is a potent activator of T‐cell adhesion, mediated by several integrins and ligands. In contrast to the effect of MnCl2, HgCl2‐induced cell adhesion probably involves an intracellular pathway. Activation of integrins by HgCl2 may play an important role in activation and migration of leucocytes involved in HgCl2‐induced immune dysregulation in vivo.
Zymosan enhances the immune response to DNA vaccine for human immunodeficiency virus type‐1 through the activation of complement system
Authors: Ara Yuki , Saito Tomoyuki , Takagi Toshitaka , Hagiwara Eri , Miyagi Yohei , Sugiyama Mitsugu , Kawamoto Susumu , Ishii Norihisa , Yoshida Takuji , Hanashi Daisuke , Koshino Tumihisa , Okada Hidechika , Okuda Kenji 
Published: March 2003

SummaryIn the present study, the adjuvant effect of zymosan on human immunodeficiency virus type‐1 (HIV‐1)‐specific DNA vaccine and the mechanism of this enhancement were studied in a murine model. We coinoculated zymosan with our candidate HIV‐1 specific DNA vaccine (pCMV160IIIB) into skeletal muscles of BALB/c mice. Higher levels of both humoral immune response and HIV‐specific delayed‐type hypersensitivity (DTH) response were observed when zymosan was coinoculated with pCMV160IIIB compared with that obtained using pCMV160IIIB alone. HIV‐specific cytotoxic T lymphocyte (CTL) activity was also enhanced. This enhancing activity was suppressed when coinoculated to the fifth complement (C5)‐deficient DDD and AKR mice. The enhanced activity was also suppressed when anti‐C3 antibody was inoculated to mice intramuscularly. There was significant induction of immunoglobulin G2a (IgG2a) and interferon‐γ (IFN‐γ) in pCMV160IIIB vaccine with zymosan. These results suggest that zymosan‐mediated DNA vaccination enhances helper T cell (Th) 1‐mediated immunity. The effect is suggested to be based on the consequences of its recruitment and activation of macrophages, dendritic cells or antigen‐presenting cells (APC) through complement activation, especially through the alternative pathway. Taken together, these results suggest that zymosan can be an effective immunological adjuvant in DNA vaccination against HIV‐1.
Tumours can act as adjuvants for humoral immunity
Authors: Brown D. M. , Fisher T. L. , Wei C. , Frelinger J. G. , Lord E. M. 
Published: March 2003

SummaryTumour cells transfected with cDNAs encoding non‐self proteins were used to investigate the ability of the immune system to respond to immunogenic antigens expressed by tumours. Secreted, intracellular and surface proteins were used as model antigens, as these reflect the potential forms of tumour antigens. Syngeneic BALB/c mice injected with viable line 1 lung carcinoma or EMT6 mammary tumour cells secreting ovalbumin (OVA) or prostate‐specific antigen (PSA) produced very high immunoglobulin G (IgG) antibody titres, equivalent to those of mice injected with protein in Freund's complete adjuvant (FCA). Secretion of the antigens was not necessary as tumour cells expressing a cell‐surface antigen (HER‐2/Neu) or an intracellular antigen – green fluorescence protein (GFP) – also generated high‐titre antigen‐specific IgG antibodies. In interleukin‐4 (IL‐4)‐deficient mice, both IgG1 and IgG2a were produced in response to OVA administered in FCA, whereas in response to tumour‐produced antigen, the antibodies switched from predominantly IgG1 to IgG2a, indicating that the mechanisms responsible for antibody induction differed between these forms of immunization. In contrast to the line 1 and EMT6 tumours, which are of BALB/c origin, OVA‐ or PSA‐producing B16 melanoma cells, which are of C57BL/6 origin, failed to elicit antibody production. This was not the result of strain differences, as a similar finding was observed when the tumours were grown in (BALB/c × C57BL/6)F1 mice, but appeared to be caused by intrinsic differences in the tumours. Furthermore, co‐injection of both B16/OVA and line 1 tumours resulted in production of anti‐OVA antibody, indicating that B16 tumours were not immunosuppressive, but instead line 1 tumours appear to exert an adjuvant effect.
Glycyrrhizin enhances interleukin‐12 production in peritoneal macrophages
Authors: Dai Ji Hong , Iwatani Yasumasa , Ishida Takaomi , Terunuma Hiroshi , Kasai Hirotake , Iwakula Yoichiro , Fujiwara Hiromi , Ito Masahiko 
Published: March 2003

SummaryInterleukin‐12 (IL‐12) is a monocyte/macrophage‐derived cytokine that plays a prominent role in the development of T helper type 1 (Th1) cell‐mediated immune responses. Glycyrrhizin (GL), an aqueous extract of liquorice root, used as Chinese medicine, is known to have various immunomodulating activities. In this study, GL showed a dose‐dependent priming effect on lipopolysaccharide (LPS)‐induced IL‐12 p40 and IL‐12 p70 (heterodimer of p40 and p35) protein production by peritoneal macrophages (PM). The maximal effect was observed when GL was intraperitoneally administered 12 hr before the PM were harvested and stimulated in vitro with LPS. The increases in IL‐12 p70 and p40 protein production were primarily due to up‐regulated transcription of IL‐12 p35 and p40 messenger RNAs (mRNAs), as demonstrated by RNase protection assay. The augmentation of IL‐12 p40 mRNA expression induced by GL pretreatment was associated with increased NF‐κB activation. Moreover, GL exhibited the same priming effect on IL‐12 production in interferon‐γ knockout (IFN‐γ–/–) mice. The production of granulocyte–macrophage colony‐stimulating factor (GM‐CSF) was not induced at any time point after GL pretreatment. These findings demonstrated the ability of GL to enhance LPS‐induced IL‐12 production by peritoneal macrophages, and indicated that the priming effect of GL on IL‐12 production was independent of both IFN‐γ and GM‐CSF.
Lipopolysaccharide and phorbol 12‐myristate 13‐acetate both impair monocyte differentiation, relating cellular function to virus susceptibility
Authors: Basta Sameh , Knoetig Sonja , Summerfield Artur , McCullough Kenneth C. 
Published: March 2003

SummaryBoth lipopolysaccharide (LPS) and phorbol 12‐myristate 13‐acetate (PMA) impeded monocyte to macrophage differentiation with respect to typical phenotypic modulation and certain phagocyte‐related processes. The down‐regulation of the porcine monocyte marker SWC1, and up‐regulation of the SWC9 macrophage marker were retarded, but not inhibited, as was the differentiation‐associated down‐regulation of p53 and myeloperoxidase. Despite this clear impairment of macrophage differentiation, not all cellular functions were equally susceptible. Both agents inhibited phagocytosis, but not low‐density lipoprotein receptor‐associated endocytosis. Only LPS inhibited tartrate‐resistant acid phosphatase up‐regulation. In contrast, increase of vacuolar acidification rates was more susceptible to PMA. The activity of certain endosomal/lysosomal enzymes – esterase, nucleotidase, peroxidase and cathepsins – was generally enhanced by both LPS and PMA. This contrasted with autophagosomal activity, detected through the induction of an antiviral state. Disruption of autophagosomes and lysosomes (methionine‐O‐methyl ester), but not lysosomes alone (glycyl‐l‐phenylalanine) reversed LPS‐induced inhibition of virus replication, without influencing the PMA‐induced antiviral effect. Thus, PMA is similar to LPS in inhibiting monocyte to macrophage differentiation, when primary blood monocytes are employed, but not all pathways are equally susceptible. The analyses demonstrate that the pathways modulated during monocyte differentiation function somewhat independently. Moreover, certain functions of monocytic cells are more important with respect to the outcome of virus infection, with autophagosomal activities in particular favouring cell survival.
Enumeration of hepatitis B surface antigen‐specific B lymphocytes in responder and non‐responder normal individuals vaccinated with recombinant hepatitis B surface antigen
Authors: Shokrgozar Mohammad A. , Shokri Fazel 
Published: March 2003

SummaryDetermination of the frequency of specific B lymphocytes has important implications for investigation of the immune response to different antigens and pathogens. Unresponsiveness to some viruses and antigens, such as hepatitis B virus (HBV) and its surface antigen (HBsAg), has been attributed to lack or insufficient production of the specific B‐cell repertoire. In this study, peripheral blood B lymphocytes of 45 adult normal individuals vaccinated with recombinant hepatitis B vaccine were transformed with Epstein–Barr virus (EBV) and cultured at different dilutions on human fetal fibroblasts as a feeder layer. The vaccinees were classified into good, poor and non‐responder groups. Following 2 to 3 weeks of incubation, culture supernatants were collected from wells containing transformed and proliferating B lymphocytes. The supernatants were subsequently screened for the presence of total immunoglobulin and antibody to HBsAg (anti‐HBs) by enzyme‐linked immunosorbent assay (ELISA). Accordingly, positive and negative wells were enumerated in each plate and the frequency of B lymphocytes producing anti‐HBs antibody was estimated based on the Poisson statistical analysis. The total number of CD19+ B lymphocytes were counted in the peripheral blood of all subjects by flow cytometry. Our results demonstrated a similar precursor frequency of specific B lymphocytes in all subject groups before vaccine administration (< 2 × 10−5). Following vaccination, however, a significant increase in the number of specific B lymphocytes was observed in good‐responder (1·5 × 10−4) and to a lesser extent poor‐responder (3·5 × 10−5) individuals, but not in non‐responders. These findings suggest a defect in either the primary B‐cell repertoire or helper T‐cell function in non‐responder individuals.