bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparative antibody response to salmonella antigens in genetically resistant and susceptible mice
Authors: XU H R , TAN Y Y , HSU H S , MONCURE C W , WANG X M
Published: July 2010

The ELISA was used to titrate the antibody response in mice inoculated with salmonella antigens. The genetically resistant A/J and susceptible C57BL/6J mice were either infected with the virulent or the avirulent Salmonella typhimurium. Alternatively, they were inoculated either once or twice with the heat-killed salmonella vaccine. No appreciable difference could be detected in the relative ability of these two strains of mice to produce antibodies against the lipopolysaccharide antigens of this pathogen under these four conditions.
Synovial fluid cells in juvenile arthritis: evidence of selective T cell migration to inflamed tissue
Authors: SILVERMAN E D , ISACOVICS B , PETSCHE D , LAXER R M
Published: July 2010

The perpetuation of chronic synovitis in juvenile arthritis (JA) is a complex interaction of local and systemic regulatory mechanism. We examined the cell surface phenotypc of synovial fluid cells and peripheral blood lymphocytes from 15 patients with J A to better understand the mechanism of local inflammation. Synovial fluid and peripheral blood mononuclcar cells were analysed for cell surface expression of CD2, CD3, CD4, CD8, CD19, CD25, CD29, CD45R and Ia using flow cytometry. We found a very low percentage of B cells with a concomitant increase of T cells in synovial fluid as compared with peripheral blood. A large percentage of the synovial fluid T cells were HLA-DR+.or activated T cells, and there was a relative decrease in CD4+ cells in synovial fluid as compared with peripheral blood. There was only a minimal increase in CD25+ synovial fluid cells. The synovial fluid CD4+ cells were mainly of the CD2high. CD29+, CD45RO phenotype. This CD4 phenotype found on synovial fluid cells from patients with JA and in particular the CD29 cell surface marker, which recognizes a common β-chain of adhesion molecules, is associated with binding to extracellular matrix proteins and is also associated with “primed” T cells. Our results demonstrated the presence of T cells which either selectively migrate to synovium and synovial fluid or are activated in situ in the joint.
Extracellular acidic pH modulates oxygen-dependent cytotoxic responses mediated by polymorphonuclear leucocytes and monocytes
Authors: GEFFNER J R , TREVANI A S , MINNUCCI F , PALERMO M S , MAUGERI N , ISTURIZ M A
Published: July 2010

In the present study, we compared the ability of human neutrophils and monocytes to display oxygen-dependent cytotoxic responses at pH 7.4 and 6.2. Our results show that cytotoxicity induced by immune complexes (IC), zymosan, N-formyl-methionyl-leucyl-phenylalanine (FMLP) and concanavalin A (Con A) were markedly increased when they were carried out at pH 6.2 instead of pH 7.4. Cytotoxicity induced by phorbol myristate acetate (PMA), on the contrary, was significantly decreased at pH 6.2. It is noteworthy that cytotoxic responses induced by IC, zymosan and Con A were also increased when, 2 h after effector cell stimulation at pH 6.2, cytotoxicity was measured at pH 7.4. Finally, when we examined possible mechanisms involved in the augmentation of cytotoxicity, we observed that the oxidative response of IC-stimulated neutrophils, measured as chemiluminescence emission, was not increased at pH 6.2, on the contrary, it was significantly decreased. The relevance of these results is discussed.
Local and systemic activationofthe whole complement cascade in human leukocytoclastic cutaneous vasculitis; C3d,g and terminal complement complex as sensitive markers
Authors: DAUCHEL H , JOLY P , DELPECH A , THOMINE E , SAUGER E , LOET X L E , LAURET H , TRON F , EONTAINE M , RIPOCHE J
Published: July 2010

We have studied complement activation both in plasma samples and In lesional skin from palients with leukocytoclaslic cutaneous vasculitis (LCV). Enzyme immunoassay (EIA) quantification of the complement activation markers. C3d,g and fhe terminal complement complex (TCC) in plasma, showed lhal their levels were significantly increased in 66% and 55% of the patienis, respectively (n = 29) compared with healthy controls, whereas the standard measurements of C3, factor B, Clq, C4and C2 were generally within normal range. Elevations of C3d,g and TCC levels in plasma were signifieantly eorrelated. Importantly, a signifieant correlation was found between the severity of the vasculitis and both C3d,g and TCC plasma levels. Immunofluorescence studies of skin biopsy specimens demonstrated simultaneous presence of peri vascular dermal deposits of C3d,g and TCC in lesional skin from 96 Mi and 80% respectively of the patients (n= 25). There was a significant eorrelation between the intensity of the deposits of bolh markers. Clusterin, a TCC inhibitory protein, was always found at the same sites of perivascular TCC deposits, Immunofluorescence studies at the epidermal basemenl membrane zone (BMZ) revealed in each case deposits of C3d,g which were accompanied by TCC deposits in 52% of the biopsy specimens. These data demonstrate that there is a local and systemic activation of the whole complement cascade in human LCV. The presenee of both C3d,g and clusterin-associatcd TCC perivascular deposits suggests an intervention of a regulatory mechanism of local complement activation in LCV. Einally, measurement of plasma C3d,g and TCC appears to be a sensitive indicator of systemic complement activation and disease severity in LCV.
Cytochrome P450IID6 recognized by LKMl antibody is not exposed on the surface of hepatocytes
Authors: YAMAMOTO A M , MURA C , DE LEMOS-CHIARANDINI C , KRISHNAMOORTHY R , ALVAREZ F
Published: July 2010

LKM1 autoantibody, directed againsl P450IID6, is accepted as a marker of a particular type of autoimmune hepatitis, but its role in the pathogenesisof the disease is controversial. Localisation of P450IID6 on the cell surface of rat hepatocytes was previously reported, suggesting that membranebound P450IID6 could be the target of LKM 1 antibodies, thus allowing immune lysis of hepatocytes. The objective of the present study was to determine, using various methods, the cell localization of P450IID6 in human and rat hepatocytes. Incubation of rat and human hepalocytes with LKMI-positive serum showed slight, if any. cell membrane staining using immunofluorcscencc. immunoperoxidase and immunoelectron microscopic sttidies. No staining of the plasma membrane of human hepatocytes was observed when incubations were earried out with imniunoaffinity-purificd antibody directed against peptide254–271, the main epitope of P450IID6 recognized by all LKMl sera tested. Chinese hamsler ovary cells, transfected with the complete P450IID6 cDNA and incubated with the supernatant from a B cell lymphoblastoid cell line prepared with the lymphocytes of a LKMI-positive patient, did not show any staining of the cell surface by immunofluorescence. Incubation of rat microsomal fraction vesicles with LKMl-positive serum, followed by prolein A-gold immunoelectron microscopy, displayed a staining of almost all vesicles, confirming that P450IID6 is present on the cyloplasmic side of the microsomal membrane, which makes it unable to be expressed on the ceil surface even if it were transported from the endoplasmic reticulum (ER). Sulpho NHS Biolin labelling of rat hepatocyte cell membranes did nol show the presence of a 50-kD molecule that could have reacted with LKMl antibody. DNA sequencing of exon 1 of the CYP2D6 gene of a patient positive for LKM l antibody did not show any difference from that of the normal published sequence of the gene. This does not favour an alteration of the NH2 terminal sequence of the P450IID6 molecule that could explain a translocation of the molecule to the luminal side of the ER, allowing its expression on the ceil surface. These results indicate that, in all likelihood. P450IID6 molecule is not present on the cell surface of normal rat and human hepatocytes. Other mechanisms than antibody-mediated cell lysis directed against membrane P450IID6 antigenic determinants must be found to account for the destruction of hepatocytes observed in this disease.
Significance of thyroglobulin antibodies cross-reactive with thyroperoxidase (TGPO antibodies) in individual patients and immunized mice
Authors: RUF J , FERRAND M , DURAND-GORDE J M , DE MICCO C , CARAYON P
Published: July 2010

Thyroglobulin (TG) and thyroperoxidase (TPO), both involved in thyroid hormone synthesis, represent major autoantigens in thyroid autoimmune disease. Despite numerous studies, the emergence, pathophysiological significance and role of autoantibodies to TG and TPO remain elusive. The recent identification of a new category of thyroid-specific autoantibody interacting with both TG and TPO (TGPO autoantibodies) offers a new opportunity in the study of thyroid autoimmunity. To gain a better insight into the significance of these TGPO autoantibodies, measurement in individual samples appeared necessary. The unique property of TGPO autoantibodies, simultaneous binding to TG and TPO, was used to set up a sandwich method which combined coated TG and radio-iodinatcd TPO. This method was found to be strictly specific for TGPO autoantibodies and sensitive enough to assay TGPO autoantibodies in serum. In humans, TGPO autoantibodies were found in most of the sera with high TG and TPO autoantibody titres, but not in sera negative for TG autoantibodies, whatever the TPO autoantibody litre. Furthermore, high TGPO autoantibody litres were found in sera strongly cytotoxic for cultured porcine thyroid cells. However, significant correlation of TGPO autoantibody titre was observed neither with TG and TPO autoantibody titres (n=48) nor with complement-dependent cytotoxicity (n= 50). TGPO antibody assay was also performed in individual plasma of CBA/J mice immunized with either human TG (n= 6) or human TPO (n= 6). Immunization with TG induced high levels of not only TG but also TGPO antibodies, which exhibited a strong reactivity for TPO and whose binding to TG and TPO was fully inhibited by TG. In contrast, immunization with TPO induced high levels of only specific TPO antibodies accompanied by low levels of specific TG antibodies. In this case TGPO antibodies were not detected. Of note, TG- and TPO-immunized mice mounted an immune response against their own TG, but did not exhibit histological signs of thyroiditis. Large panels of TG and TPO MoAbs were also investigated with this method: 18/25 TG MoAbs and only 1/13 TPO MoAbs were found cross-reactive. Taken together, these data provide evidence that TGPO antibodies are effectively present in individual patients and TG-immunized mice, are different from specific TG and TPO antibodies, and may derive from natural B cell repertoire by autoimmune processes involving TG and not TPO.
On the association between amyloid fibrils and glycosaminoglycans; possible interactive role of Ca2+ and amyloid P-component
Authors: STENSTAD T , MAGNUS J H , SYSE K , HUSBY G
Published: July 2010

We have previously reported the specific association of glycosaminoglycans (GAG) and proteogly-cans (PG) with amyloid fibrils and characterized the polysaccharides directly extracted from amyloid-laden tissues. In the present study we further elucidate the association between purified amyloid fibrils and GAG/PG with special reference to those GAG/PG associated with amyloid P-component (AP) and the interactive role of Ca2+ ions. Amyloid fibrils were isolated from human hepatic AA amyloid employing water extraction with and without preceding removal of AP. an extrafibrillar protein component of all amyloids, using sodium citrate. GAG/PG co-isolated with the amyloid extracts, with and without AP, were isolated and characterized. Agarose-affinity chromatography of extracts containing AP was performed, and the GAG associated with this extrafibrillary protein were characterized as well. Several different GA/GPG populations were demonstrated in the various extracts. The abolition of calcium-dependent binding markedly influenced the amount of GAG/PG recovered in the fibril extracts, as well as the total amount of amyloid material obtained. Thus, it seems that calcium plays an important role in the association between the fibrils and the sugar moieties, and that a significant fraction of the GAG found in amyloid exhibits a Ca2+-dependent fibril-GAG interaction. No significant difference in the proportion between galactosaminoglycans and glucosamines was, however, disclosed when the two extraction protocols were cbinpared, suggesting that no particular GAG species has a higher affinity for the fibrils themselves. Both dermatan/chondroitin sulphate and heparan sulphate identified in the present study exhibited a Ca2+ -dependent interaction with AP, supporting previous findings. However, the amyloid-associated galactosaminoglycans found, especially the large PG appearing in small amounts, seemed to have a higher affinity for the extrafibrillar AP than the other GAG.
Higher anti-heparan sulphate reactivity during systemic lupus erythematosus (SLE) disease exacerbations with renal manifestations; a long term prospective analysis
Authors: KRAMERS C , TERMAAT R M , ter BORG E J , van BRUGGEN M C J , KALLENBERG C G M , BERDEN J H M
Published: July 2010

Cross-reactive antibodies against heparan sulphate (HS) have been suggested to play a role in initiating renal disease in SLE. Recently, we found that HS-reactivity is mediated by anti-DNA antibodies complexed with DNA and histones. To evaluate the clinical significance of anti-HS reactivity, we studied prospectively a cohort of 72 consecutive SLE patients, of whom 22 experienced 40 exacerbations. In 20 of these exacerbations renal symptoms were present. In these 20 exacerbations significantly higher anti-DNA (median 1:160) and anti-HS (median 1:30) titres were detected compared with exacerbations without renal manifestations (median 1:60 for anti-DNA and negative for anti-HS). There were no correlations with other symptoms of SLE. Anti-HS titres showed a significant correlation with anti-DNA antibody titres (rs= 0·57, P < 0·05). Anti-HS without anti-DNA reactivity was never detected. Some SLE patients showed a high anti-DNA titre without anti-HS reactivity, suggesting that not all anti-DNA antibodies are able to bind to histone/DNA complexes and thus to exhibit anti-HS reactivity. Our findings indicate that anti-HS reactivity is correlated with renal disease in SLE.
Dissociation of specific and total IgE antibody responses following repeated low-level infections withNippostrongylus brasiliensisin rats
Authors: YAMADA M , UCHIKAWA R , NAKAZAWA M , ODA M , ARIZONO N
Published: July 2010

IgE, IgG and mast cell responses were studied in rats infected weekly with 10 larvae of Nippostrongylus brasiliensis (NB). Worm recovery at 8 weeks of repeated infections was six-fold greater than that of a single infection with 10 larvae, suggesting t he accumulation of worms during the repeated infections. Total serum IgE was increased after 2 weeks of infection, and further increased after repeated infections: at 6 weeks of infection the level was four to six times higher than that after a single infection. Anti-NB IgG1 levels were also significantly higher after repeated infections than after a single infection. On the other hand, there was no significant difference in the level of anti-NB IgE between single and repeated infections, as determined by ELISA, as well as by passive cutaneous anaphylaxis (PCA) reaction. Mastocytosis was induced in the small intestine after both single and repeated infections, but the levels did not differ between the two These results indicate that total IgE and specific IgG1 production are augmented by repeated helminth infections, but specific IgE and mast cell responses are not. This pattern of response may minimize the development of IgE-dependent hypersensitivity reactions with repeated helminth infections.
Cell-mediated pathology during murine malaria-associated nephritis
Authors: LLOYD C M , WOZENCRAFT A O , WILLIAMS D G
Published: July 2010

We have studied the cellular mechanisms involved in the development of nephritis during acute and chronic murine malaria infections induced by Plasmodium vinckei petteri and P. berghei respectively. Albuminuria and uraemia were observed during the early stages of both types of infeclio, and were associated with glomerular and interstitial hypercellularity. There was a gradual increase in numbers of CD45+ cells from the early stages of both infections onwards. These infitrates contained CD4+ and CDS cells and mononuclear phagocytes. The interstitial and glomerular hypercellularity was due to an influx of inflammatory cells rather than an increase in renal cell division. These findings indicate the importance of cell-mediated immune mechanisms in the development of nephritis during murine malaria and illustrate an example of naturally occurring infection-induced nephritis.
Increased proportion of responders to a murine anti-CD3 monoclonal antibody of the IgG1 class in patients with systemic lupus erythematosus (SLE)
Authors: BLASINI A M , STEKMAN I L , LEON-PONTE M , CALDERA D , RODRIGUEZ M A
Published: July 2010

A group of Venezuelan patients with SLE showed an increased proportion of responders to Leu-4, an anti-CD3 Mo Ab of the IgG1 class, compared with ethnically matched non-SLE patients and healthy controls. The rate of proliferative responses or IL-2 production induced by Mo Ab Leu-4, and the helper effect of macrophages from Leu-4 responders on T cells from a third-party donor were comparable in patients and controls. No significant differences in the binding of murine IgG1 molecules by macrophages from SLE patients and controls were observed. The proportion of monocytes/macrophages expressing FcγRI was significantly higher in SLE patients. However, the expression of FcRII, the type capable of supporting Leu-4-mediated responses, and of FcγRIII was comparable in monocytes from SLE patients and controls. Our results suggest that Venezuelan patients with SLE may have a genetic predisposition for the expression of the phenotypic variant of FcγRII capable of binding murine IgG1 molecules.
Induction of neutrophil respiratory burst by tumour necrosis factor-alpha; priming effect of solid-phase fibronectin and intervention of CD 11b-CD18 integrins
Authors: DAPINO P , DALLEGRI F , OTTONELLO L , SACCHETTI C
Published: July 2010

Human neutrophils, added to fibronectin (FN)-coated polystyrene wells and exposed to tumour necrosis factor-alpba (TNF-α, were found to exhibit a prolonged production of superoxide anion (Q2) after a lag period of approx 30 min. The O2 production, but not the cell adherence to FN, was completely inhibited by two MoAbs against CD18 and by a MoAb against CD11b, suggesting the involvement of CD11b-CD18 integrins in the neutrophil oxidative response. When neutrophils were induced to adhere to FN by incubation for 30 min on FN-coated surfaces and then washed to remove non-adherent cells, FN-anchored cells exhibited a rapid onset of O2 production in response to TNF-α. This suggests that FN primes neutrophils for the TNF-α-mediated respiratory burst. The O2− production by adherent neutrophils could be inhibited by anti-CD11b and anti-CD18 MoAbs only when the MoAbs were present both during the induction of adherence and during the subsequent exposure of FN-bound cells to TNF-α. The incapacity of MoAbs, added to neutrophils during the induction of adherence, to modify the characteristics of the subsequent neutrophil response to TNF-α suggests that the FN-mediated cell priming is independent of the interaction of CD11b-CD18 integrins with the FN substrate. The results are consistent with the intervention of three classes of cell receptors in the TNF-α-induced oxidative burst of neutrophils plated on FN: (i) neutrophil FN-binding sites, distinct from CD11b-CD18 and responsible for the cell priming; (ii) CD11b-CD18 integrins, absolutely required for permitting the cell triggering; and (iii) TNF-α receptors, responsible for switching on a rapid cell response in primed cells. The requirement of multiple classes of receptors for the full expression of the cell function can be envisaged as a natural precautionary measure to control the neutrophil responsiveness to TNF-α and, in turn, the TNF-α-dependent neutrophil-mediated oxidative injury at sites of inflammation.
Early detection of the allogeneic effect in the mixed lymphocyte reaction (MLR) by anti-CD3 IgA mouse monoclonal antibody
Authors: MANZO C , FERRAIUOLO C
Published: July 2010

MLR is a 6-day routine proliferation assay used to demonstrate HLA disparity between two individuals. However, on day 2 only a negligible proliferation can be detected. In contrast, we found a significant increase of the extent of proliferation if an anti-CD3 IgA antibody was present in the culture. Since the CD3 proliferation pathway cannot be triggered by an antibody of IgA isotype the result was interpreted as a consequence of a synergism between the allogeneic stimulus and the anti-CD3 antibody effect. The level of this proliferation well correlates with the HLA difference, thus suggesting the use of this methodology for a rapid detection of the allogeneic reaction.
In vivoexpression of human immunoglobulin germ-line mRNA in normal and in immunodeficient individuals
Authors: ISLAM K B , BASKIN B , CHRISTENSSON B , HAMMARSTRÖM L , SMITH C I E
Published: July 2010

Previous in vitro studies suggest that transcription of the unrearranged immunoglobulin switch region and its 5′ flanking region precedes isotype switching. These transcripts, which arc devoid of a variable region, contain unique exons and are called germ-line (GL) mRNA. A crucial point in this regard is whether such transcripts could be detected in vivo, and if their expression correlates with immunoglobulin class switching in health and disease. To understand the in vivo role of this transcriptional activity we have adapted the reverse transcription-polymerase chain reaction (PCR) to analyse the GL transcripts from unstimulated peripheral blood mononuclear cells (PBMC) in healthy individuals and in different immunological diseases. Furthermore, mononuclear cells from different human organs were also analysed. We report here that GL (Iα. Iγ and Iɛ used lo designate the GL mRNA for IgA. IgG and IgE, respectively) nRNA are expressed differentially during ontogeny of B cells. Unexpectedly, no difference of Iα mRNA expression between the PBMC and the secondary lymphoid organs was detected. Rather. the levels of GL transcripts were correlated to ihe number of sIgM+ cells. GL mKNA of all three isotypes could be detected in PBMC from healthy donors, whereas there was a decrease of specific GL transcript synthesis in individuals with Immunoglobulin deficiency. Furthermore, during the in vivo immune response in a parasitic infection, we could demonstrate an induction of GL k mRNA during in vivo immune response. Concomitantly. there was also increased synthesis of productive F. transcripts. These findings implicate a potential role of GL transcription during in vivo immunoglobulin class switching.
Hereditary angioneurotic oedema: characterization of plasma kinin and vascular permeability-enhancing activities
Authors: SHOEMAKER L R , SCHURMAN S J , DONALDSON V H , DAVIS A E
Published: July 2010

The mediator(s) responsible for localized enhanced vascular permeability that characterizes an exacerbation of hereditary angioneurotic oedema (HAE) is thought to be a product of either contact or complement system activation. In contrast to normal individuals, plasma from these patients generates both kinin and vascular permeability-enhancing activity following incubation at 37°C. Depletion of C1 inhibitor in both normal and C2-deficicnt plasma, but not in contact factor-deficient plasmas, resulted in generation of these activities. The kinin activity from incubated HAE plasma was susceptible to kininase inactivation and was blocked by a Bk2 receptor antagonist. Furthermore, this activity was isolated from HAE plasma; amino acid sequence analysis proved it to be bradykinin. Similarly, the vasopermeability-enhancing activity from ethanol-fractionated or boiled HAH plasma, collected during either attack or remission, co-eluted with bradykinin on reverse-phase high performance liquid chromatography (HPLC). These studies conclusively demonstrate that bradykinin is the major kinin and mediator of enhanced vascular permeability generated during incubation of HAE plasma. The role of other bioactive products, such as the C2 kinin, at local sites of oedema formation remains lo be further defined.
Human TcRγδ+ lymphocyte response on primary exposure toPlasmodium falciparum
Authors: ROUSSILHON C , AGRAPART M , GUGLIELMI P , BENSUSSAN A , BRASSEUR P , BALLET J J
Published: July 2010

In 29 patients experiencing their first P. falciparum malarial attack. blood levels of TcRγδ+ lymphocytes were studied from the onset of infection to up to 6-9 months later. Blood TcRγδ+ lymphocytes, revealed using the TcRδ1 monoclonal antibody (MoAb) were increased both in absolute and relative numbers. Alterations lasted for up to 3-4 months following the attack. A TiγA/ BB3 reactive Vγ/9 subset was preferentially amplified, in vitro, TcRγδ+ lymphocytes from both malaria-sensitized and unprimed donors responded to P. falciparum schizont extract (PFSE). PFSE-stimulated polyclonal T cell lines consisted principally in TcRγδ+ cells with a TiγAd+/BB-3+ phenotype. Several TcRγδ+ T cell clones obtained from patients recovering from acute malarial attack were maintained in the presence of PFSE and autologous irradiated PBL. They belong to the Vγ9 subset. In long-term cultures. TcRγδ+ clones progressively lost their capacity to react to PFSE antigen while they were able to proliferate and to exert cytotoxic activity in response to autologous TcRγδ+, PFSE-specific T lymphocyte clones. This suggests that regulatory interactions occur between activated TcRγδ+ and TcRγβ+ cells generated by P. falciparum. Sequential variations in blood TcRγδ+ and TcRγβ+ lymphocyte levels after primary exposure to P. falciparum suggest that such regulatory interactions may occur in vivo.
In vivotreatment with a monoclonal antibody to T helper cells in experimental autoimmune glomerulonephritis in the BN rat
Authors: REYNOLDS J , PUSEY C D
Published: July 2010

Experimental autoimmune glomerulonephritis (EAG) was induced in brown Norway (BN) rats by a single i.m. injection of homologous glomerular basement membrane (GBM) in Freund's complete adjuvant. This model of anti-GBM disease is characterized by the development, over several weeks, of circulating and deposited anti-GBM antibodies, accompanied by albuminuria. We examined the effects of treatment with MoAb W3/25 (anti-CD4) at different doses, starting at the time of immunization and continued for the duration of the study or for a limited period only. Continued treatment with W3/25, at a dose of 5 or 10 mg/kg intraperitoneally three times per week for 4 weeks. produced a marked reduction in circulating anti-GBM antibodies, absence of detectable deposited antibody and virtual absence of albuminuria. When W3/25 treatment, at 5 or 10 mg/kg, was stopped after 2 weeks, there was still a significant reduction in anti-GBM antibodies and albuminuria at 4 weeks. A similar effect on the disease was achieved when W3/25 was administered only three times during the first week at a dose of 30 mg/kg. Animals injected with W3/25 at a dose of 10 mg/kg through the course of disease showed < 10% W3/25+ cells by KACS analysis of splenic lymphocytes at week 4, while controls and animals treated for shorter periods showed >30% W3 25+ cells. These results demonstrate that W3/25 can prevent the development of EAG and that this effect is not dependent on persistent depletion of T cells. Further work is necessary to determine whether anti-T cell therapy is effective in established EAG and may be worth investigating in human anti-GBM disease.
Production of tumour necrosis factors by human T cells stimulated by a superantigen, toxic shock syndrome toxin-1
Authors: AKATSUKA H , IMANISHI K , INADA K , YAMASHITA H , YOSHIDA M , UCHIYAMA T
Published: July 2010

The capacity of human T cell subsets, CD4+ or CD8+ T cells, to produce tumour necrosis factors (TNF-α and TNF-β) upon stimulation with toxic shock syndrome toxin-l (TSST-I) and the requirement for MHC ctass II molecules on accessory cells (AC) in the response were investigated. The capacity of CD4+ T cells was much higher than that of CD8+ T cells in TSST-1-induced production of TNF-α and TNF-β. The expression of MHC class II molecules on AC was required in the response.
Intraglomerular expression of transforming growth factor-beta 1 (TGF-β1) mRNA in patients with glomerulonephritis: quantitative analysis by competitive polymerase chain reaction
Authors: IWANO M , AKAI Y , FUJII Y , DOHI Y , MATSUMURA N , DOHI K
Published: July 2010

TGF-β1 is involved in the pathogenesis of glomerular sclerosis. We studied the intraglomerular expression of TGF-β1 mRNA in patients with glomerulonephritis using competitive polymerase chain reaction (PCR). This method is sensitive enough to quantify cDNA copies of mRNA present in small amounts of samples. Renal biopsy specimens were obtained from 42 patients with various kinds of glomerulonephritis. Ten glomeruli were dissected from renal biopsy specimens. Normal glomeruli were also obtained from the resected kidneys of eight patients with renal cell cancer. Total RNA was extracted from the glomeruli and reverse transcribed into cDNA with reverse transcriptase. To prepare samples containing identical amounts of β-actin cDNA (8 pg), we performed competitive PCR by co-amplifying mutant templates of β-actin with a unique EcoRI site. Next, to measure TGF-β1 cDNA, we performed competitive PCR by co-amplifying mutant templates of TGF-β1. We observed a higher glomerular expression of TGF-β1 mRNA in cases of mesangial proliferative glomerulonephritis having a moderate increase in mesangial matrix, diabetic nephropathy and diffuse proliferative lupus nephritis, compared with normal glomeruli. Results suggest that the intraglomerular synthesis of TGF-β1 may be involved in the progression of glomerulonephritis in humans.
Expression of the gene for inducible nitric oxide synthase in experimental glomerulonephritis in the rat
Authors: COOK H T , EBRAHIM H , JANSEN A S , FOSTER G R , LARGEN P , CATTELL V
Published: July 2010

Nitrite, a stable product of nitric oxide (NO), is synthesized in vitro by glomeruli in experimental glomerulonephritis. We have now studied the expression of the gene for inducible NO synthase (iNOS) in accelerated nephrotoxic nephritis (NTN). The purpose of the study was to confirm in vivo induction of iNOS in this model of immune complex disease, and to relate the onset of induction and the level of expression to pathogenic events in the model. Glomeruli from rats with NTN were isolated at 6 h, 24 h and 2, 4 and 7 days and total RNA extracted. RNA (10/μg) was reverse transcribed and polymerase chain reaction (PCR) was performed with primers homologous to rat vascular smooth muscle iNOS and rat β actin. A 222-base PCR product corresponding to iNOS mRNA was present in all experimental animals. iNOS expression was also found in activated macrophages, neutrophils and IL-1-stimulated but not unstimulated mesangial cells. Quantitative competitive PCR was carried out on glomerular samples using a 514-bp mutant of a 735-bp PCR product. iNOS expression was present at low levels in normal glomeruli and was markedly enhanced at 6 h after the induction of glomerulonephritis and peaked at 24 h. Increased iNOS expression persisted to day 7. β actin mRNA levels were similar in all glomerular specimens. This study demonstrates that there is in vivo induction of iNOS in immune complex glomerulonephritis, corresponding to the generation of nitrite we have previously reported. iNOS gene expression is detectable within 6 h of induction of NTN, indicating the onset of gene transcription is closely related lo the initial formation of immune complexes.
Expression of the chemokine superfamily in rheumatoid arthritis
Authors: HOSAKA S , AKAHOSHI T , WADA C , KONDO H
Published: July 2010

The infiltration of leucocytes into the Joint of rheumatoid arthritis (RA) is believed to be mediated by chemotactic factors released by activated cells. In this study, examination was made of the gene expression and production of the chemokine superfamily in RA patients by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoprecipitation. Cultured synovial fibroblasts were found capable of expressing and producing IL-8, GRO, monocyte chemotactie and activating factor (MCAF), macrophage inflammatory protein-1α (MIP-1α), MIP-1β and RANTES in response to IL-1α, The expression of IL-8, GRO. MCAF, MIP-1α, and MIP-1β was clearly shown to increase in freshly isolated synovial fluid mononuclear cells (SFMC) of RA patients, in contrast to peripheral blood mononuclear cells (PBMC) of RA patients and normal subjects. The gene expression of RANTES appeared to be the same for RA SFMC. RA PBMC, and normal PBMC. Thus, the over-expression of various chemokines may promote the recruitment of inflammatory cells into rheumatoid inflamed joints.
Spontaneous expression of IL-4 mRNA in lymphocytes from children with atopic dermatitis
Authors: TANG M L K , KEMP A S
Published: July 2010

Normal lymphocytes do not generally produce or secrete lymphokines in the resting or unstimulated state and only express or release cytokines following activation. Recently, the spontaneous production of intracellular interferon-gamma (IFN-7) and spontaneous secretion of IL-6 has been documented in patients with atopic dermatitis. These findings indicated that lymphocytes had been previously activated in vivo. Such in vivo activation may also be associated with spontaneous production of IL-4. As measurement of IL-4 secretion by immunoassay is complicated by poor sensitivity, and only provides information on the net amount of cytokine present after secretion, adsorption, consumption and degradation have occurred, IL-4 mRNA expression in peripheral blood lymphocytes from children with atopic dermatitis and controls was examined by polymerase chain reaction (PCR)-assisted mRNA amplification. Spontaneous expression of IL-4 mRNA was detected in four of eight patients with severe atopic dermatitis. Following stimulation in vitro, seven of eight atopic patients demonstrated detectable IL-4 mRNA. In comparison, no spontaneous expression of IL-4 mRNA was found in controls, and only six of 10 controls expressed IL-4 mRNA in stimulated cultures. The spontaneous expression of IL-4 mRNA in unstimulated cultures from children with atopic dermatitis supports the possibility that previous in vivo activation has occurred, and suggests that IL-4 production is increased in vivo in atopic dermatitis. This in vivo activation together with the constitutive expression of IL-4 mRNA are likely to contribute to the spontaneous in vitro production of IgE in atopic patients.
Microglial cells qualify as the stimulators of unprimed CD4+ and CD8+ T lymphocytes in the central nervous system
Authors: CASH E , ROTT O
Published: July 2010

The potential of central nervous system (CNS)-derived cells for initiating T cell responses is not known. Using the capacity of unprimed T cells to respond to allogeneic determinants on antigen presenting cells (APC). we assessed the ability of microglial cells lo act as stimulators of primary T cell responses in vitro. For this purpose, microglial cells were activated with lipopolysaccharide (LPS). interferon-gamma (IFN-γ), or by phagocytosis of progenitor oligodendrocytes and subsequently tested for their ability to induce a proliferative response of naive, resting T cells. Activated microglial cells induced a significant proliferation of virgin, alloreactive CD4+ and CD8+ T lymphocytes, with a more substantial response of highly purified CD4+ than of CD8+ expressing T cells. Phagocytosis activation was the most efficient stimulus to induce this APC competence on microglial cells. By contrast. IFN-γ pretreated. MHC-expressing astrocytes were unable to induce similar responses of alloreactive CD4+ or CD8+ T cells under the same experimental conditions. Collectively, our data suggest the role of activated microglia as the fully immunocompetent accessory cell population of the CNS.
Persistence of virus and viral genome in myocardium after Coxsackievirus B3-induced murine myocarditis
Authors: RABAUSCH-STARZ I , SCHWAIGER A , GRÜNEWALD K , MÜLLER-HERMELINK H-K , NEU N
Published: July 2010

Following infection with Coxsackievirus B3 (CVB3). A-strain mice develop ongoing myocarditis that persists after the virus ceases to be cultivatable from heart tissue. We studied the natural history of this virus-induced but apparently autoimmune inflammation by means of in situ hybridization (ISH) and by polymerase chain reaction (PCR). Both ISH and culture allowed detection of virus up to 2 weeks post-infection in virtually all heart tissues. In contrast, PCR revealed the presence of viral genome for a substantially longer period of time, i.e. at least 34 days after CVB3 infection. Similarly, the majority of mice showed myocardial inflammation at this time point. However, the persistence of virus did not correlate with ongoing myocarditis, and vice versa. Most mice with ongoing myocarditis produced heart myosin autoantibodies, most probably as a result of tissue damage. The lack of correlation between presence of ongoing inflammation and persistence of virus supports our previous view that the late phase of CVB3-induced myocarditis is mediated by auto immunological mechanisms.
Echinococcus multilocularis: parasite-specific humoral and cellular immune response subsets in mouse strains susceptible (AKR, C57BI/6J) or ‘resistant’ (C57B1/10) to secondary alveolar echinococcosis
Authors: GOTTSTEIN B , WUNDERLIN E , TANNER I
Published: July 2010

Parasite-specific humoral and cell-mediated immune responses were investigated in highly susceptible (AKR and C57B1/6J) and relatively resistant (C57B1/10) mice undergoing secondary alveolar echinococcosis (infection with Echinococcus multilocularis metacestode). The parasite-specific proliferative immune response of lymph node cells upon in vitro antigen stimulation remained weak in all three mouse strains. By day 30p.i., CD4+ lymphoblast cells dominated the total population of blast cells in all three mouse strains. There was, however, an unexpectedly high proportion of CD8+ blast cells; by day 90p.i., a marked proportional increase in CD8+ cells was seen in susceptible (AKR and C57B1/6J), but not in resistant (C57B1/10) mice. Susceptible, but not resistant mice exhibited a significantly decreased responsiveness of lymph node ceils to concanavalin A (Con A) stimulation on day 90 p.i. Analysis of the humoral immune response by ELISA showed that resistance in C57B1/10 mice was associated with the ability of the host to synthesize antibodies to Em2 of the IgG3 and IgG1 isotype. Em2 is a lectin-binding carbohydrate antigen of the laminated layer. In susceptible AKR and C57B1/6J mice, low levels of antl-Em2 antibodies of the IgG2a isotype were detected. Anti-Em2 antibodies of the IgG3/lgG1 isotype, however, were absent. Differences in subclass-specific IgG responses were confirmed by immunoblot analyses. Our findings suggest that differences in antigen recognition (with respect to subsets of humoral and cellular immune components), probably controlled by non-H-2 gene(s), coupled to immune suppression modulated by CD8+ cells and/or respective cytokines, may determine susceptibility or resistance in experimental infection with E. multilocularis.
Dipeptidyl peptidase IV (DP IV) activity in serum and on lymphocytes of MRL/Mp-lpr/lprmice correlates with disease onset
Authors: KUBOTA T , IIZUKA H , BACHOVCHIN W W , STOLLAR B D
Published: July 2010

DP IV (CD26), a serine protease expressed on activated T cells, participates in immune responses in vivo as well as in vitro. We measured cell surface and serum DP IV in mice of the autoimmune MRL/Mp-lpr/lpr (MRL/1) strain, which is characterized by massive T cell proliferation and production of anti-nuclear autoantibodies. The mass of inguinal lymph nodes correlated with scrum DP IV activity. Furthermore, serum DP IV activity increased markedly in parallel with the acceleration of lymph node swelling and anti-nDNA antibody production. Serum DP IV activity in 16-week-old MRL/I mice reached levels up to three times higher than those in age-matched MRL/Mp-+/+ mice or BALB/c mice. Immunohistochemical staining and flow cytometric analysis identified DV IV on surfaces of lymphocytes from the enlarged lymph nodes of MRL/1 mice. Subcutaneous injection of the mechanism-based inhibitor, Pro-boroPro, reduced protease activity in serum and cell suspensions prepared from spleen and lymph nodes, confirming the identity of the enzyme as DP IV. These results indicate that the massively accumulating lymphocytes of MRL/1 mice have a property characteristic of activated T cells, although they express little surface CD4 or CD8 and do not produce IL-2. DP IV may participate in the role these cells play in the pathogenesis of MRL/1 autoimmune disease.
Increase of circulating γ/δ T lymphocytes in the peripheral blood of patients affected by active inflammatory bowel disease
Authors: GIACOMELLI R , PARZANESE I , FRIERI G , PASSACANTANDO A , PIZZUTO F , PIMPO T , CIPRIANI P , VISCIDO A , CAPRILLI R , TONIETTI G
Published: July 2010

In order to study the role of γ/δ T cells in the pathogenesis of inflammatory bowel disease (IBD) in humans, we measured the percentage of these cells in the peripheral blood, assessed the ratio of the non-disulphide-linked (δTCS1) type of T cell receptor (TCR) in the total γ/δ T cells, studied the co-expression of γ/δ TCR and accessory molecules CDS and CD 16, and compared these data with both the type and the activity of the disease. Percentage levels and absolute numbers of γ/δ+ T cells were higher in active patients than in controls (P < 0·05), mainly as a result of an increase of Vδ1+ (δTCS1) T cell subset (P < 0·05). This trend was strongly retained independently of disease activity and clinical picture. An increased percentage of TCRδ1+/CD16+ cells was observed in our patients compared with controls (P < 0·05). In contrast, no difference was observed as far as the TCRδ1+/CD8+ cells were concerned. These results suggest that IBD is associated with an expansion of γ/δ T cells in peripheral blood, which may play a role in the pathogenesis of these disorders.
Envelope glycoprotein (gp120) from HIV-1 enhancesMycobacterium aviumgrowth in human bronchoalveolar macrophages; an effect mediated by enhanced prostaglandin synthesis
Authors: DENIS M
Published: July 2010

Human bronchoalveolar lavage (BAL) macrophages were obtained from normal human volunteers and infected with an AIDS-associated strain of Mycobacterium avium. infected cells were exposed to purified envelope glycoprotein (gp120) from HIV-1 or to the recombinant non-glycosylated gp120 fragments PBI-RF and PBI-IIIB. Native gp120 increased Myco. avium growth in human cells from six separate donors, whereas the non-glycosylated fragments of gp120 had no such effect. Moreover, gp120 induced a substantial secretion of prostaglandin E2 (PGE2) from macrophages; inclusion of indomethacin blocked the enhanced permissiveness of infected cells treated with gp120. Soluble CD4 also neutralized the effect of gp120. Overall, these results indicate a role for gp120 in the susceptibility of AIDS patients to Myco. avium infections, mediated by an enhanced PGE2 release.
Brain-reactive autoantibodies in BB/d rats do not recognize glutamic acid decarboxylase
Authors: DAVENPORT C , LOVELL H , JAMES R F L , TODD I
Published: July 2010

The BB rat spontaneously develops insulin-dependent diabetes mellitus (IDDM) similar to that in humans. The most practical markers of β cell autoimmunity are circulating antibodies to islet cell components. In particular autoantibodies to the enzyme glutamic acid decarboxylase (GAD) are a common feature of IDDM development in humans. This study aims at investigating the prevalence and levels of autoantibodies in BB rats to antigens in a semipurified, GAD-enriched preparation from rat brain. Eighteen diabetes-prone BB/d rats (10 male and eight female) were tail bled weekly from age 28 days to 113 days and antibodies detected on the rat brain preparation by ELISA. Antibody levels were expressed as arbitrary units relative to a standard positive serum. Individual rats varied in the time and order of antibody appearance and IDDM onset, with the earliest occurrence being 42 days and 69 days, respectively. In some rats antibody production was maintained but declined in others. By 113 days 85% of diabetic rats had at some time been positive for autoantibodies to brain components, compared with 25% of non-diabetics (P= 0·09 by Fisher's exact test), Immunoabsorption studies using recombinant rat GAD-65 or recombinant human GAD-67 failed to inhibit the binding of BB rat sera to the original rat brain preparation. A capture ELISA using GAD-6 MoAb to capture GAD-65 from rat brain preparation or from a preparation of recombinant rat GAD-65, failed to detect anti-GAD antibodies in BB rats. Immunofluorescent staining of tissue sections showed the autoantibodies to be brain-specific, but having distinct staining patterns to the anti-GAD antibodies of Stiff Man Syndrome serum. In conclusion, BB rats possess autoantibodies reactive with rat brain antigens which may he associated with IDDM. However, these are not directed against GAD.
T cell lines recognizing the 70-kD protein of U1 small nuclear ribonucleoprotein (U1snRNP)
Authors: Fenning S , Wolff-Vorbeck G , Hackl W , Krawinkel U , LÜHrmann R , Northemann W , Peter H H , Schlesier M
Published: July 2010

In sera of patients with mixed connective tissue disease (MCTD) high titres of IgG autoantibodies to U1snRNP-specific proteins (70kD, A, C) are found, suggesting an antigen-driven and T cell-dependent process. In order to establish U1snRNP-specific T cell lines we cultured under various culture conditions mononuclear cells from MCTD patients and healthy donors with a highly purified UsnRNP preparation from HeLa cells. Nine T cell lines were established by limiting dilution cloning from two MCTD patients and five T cell lines from a healthy individual. All T cell lines expressed the TCRαβ/CD3 complex. Surprisingly, most of the T cell lines exhibited the CD8 phenotype. Irrespective of this phenotype, all T cell lines showed a proliferative response to an N-terminal pan (aa 51–195) of recombinant U1-specific 70-kD protein. One CD8+ T cell clone exhibited cytotoxic activity against an autologous B cell line pulsed with snRNP or recombinant fragments (aa 51–195 and aa 51–88). Interestingly, two T cell lines proliferated in response to four recombinant polypeptides representing different parts of the U1snRNP 70-kD protein. Since regions of sequence homology are distributed over the 70-kD molecule, it is suggested that conserved motifs may be recognized by the T cell lines.