bluuebunny/crossref_metadata_2025 huggingface dataset:
Anti-CD28 antibody- and IL-4-induced human T cell proliferation is sensitive to rapamycin
Authors: LUO H , CHEN H , DALOZE P , ST-LOUIS G , WU J
Published: July 2010

Rapamycin (RAPA) is a potent immunosuppressant. In this study we investigated the effect of RAPA on T cell proliferation triggered by various stimuli in an in vitro human model. The proliferation of T cells stimulated via an alternative pathway using phorbol myristate acetate (PMA) and anti-CD28 antibody (αCD28) in the absence of antigen-presenting cells (APC) was strongly inhibited by RAPA. T cell proliferation provoked via a combination of CD3/TCR and CD28 pathways using anti-CD3 antibody (αCD3) plus αCD28 was also inhibited by RAPA in the presence of APC. The mitogen (phytohaemagglutinin (PHA) or αCD3)-induccd up-regulation of expression of the IL-2 receptor a chain (IL-2Rα) and the IL-4 receptor (IL-4R) was sensitive to RAPA. This suggests that RAPA's interference with the IL-2 and IL-4 autocrine loops during T cell activation might contribute to RAPA's overall immunosuppressive effect. We have further demonstrated in a two-stage culture system that RAPA strongly inhibited IL-4-stimulated proliferation of T cells, the latter being either pretreated with αCD3 in the presence of APC, or with PMA plus αCD28 in the absence of APC. The result suggests that the Ca++ influx during the pretreatment is not obligatory for T cells to achieve IL-4 responsiveness. The results also indicate that RAPA's antiproliferative effect on IL-4-stimulated T cells is not contingent on the various mechanisms of cell priming. Therefore, RAPA's major target is probably at the second stage after the priming. Our study has extended current knowledge about the effect of RAPA on human T cells.
A high proportion of the Vδ1+ synovial fluid γδ T cells in juvenile rheumatoid arthritis patients express the very early activation marker CD69, but carry the high molecular weight isoform of the leucocyte common antigen (CD45RA)
Authors: KJELDSEN-KRAGH J , QUAYLE A J , VINJE O , NATVIG J B , FØRRE Ø
Published: July 2010

We have previously shown that γδ T cells in the synovial compartment of patients with juvenile rheumatoid arthritis (JRA) express activation antigens (CD69 and HLA-DR) and that they are predominantly of the Vδ1 subset. In this study we have analysed the expression of activation antigens (CD69 and HLA-DR) and different isoforms of the leucocyte common antigen (CD45RO and CD45RA) on the Vδ1 and the V52 subsets of γδ T cells in paired samples of synovial fluid and peripheral blood of nine patients with JRA, and in the peripheral blood of five children with idiopathie seoliosis. In the synovial fluid of children with JRA, there were significantly more Vδ1+CD69+ and Vδ2+CD69+ cells compared with the peripheral blood of the same patients. In contrast, however, in the synovial fluid the Vδ1 and the Vδ2 subsets differed with respect to the expression of the two isoforms of the leucocyte common antigen. The majority of the Vδ1+ cells expressed the high molecular weight isoform (CD45RA+) while most of the Vδ2+ cells carried the low molecular weight variant (CD45RO+) of this molecule.
Reduction of monocyte-macrophage activation markers upon anti-CD4 treatment. Decreased levels of IL-1, IL-6, neopterin and soluble CD14 in patients with rheumatoid arthritis
Authors: HORNEFF G , SACK U , KALDEN J R , EMMRICH F , BURMESTER G R
Published: July 2010

Anti-CD4MoAbs have been successfully used in initial treatment trials of rheumatoid arthritis. One remarkable feature of this therapy was the early reduction of synovitis along with a decrease of the erythrocyte sedimentation rate (ESR) and the C-reactive protein (CRP). Since not only T helper celis. but also monocytes-maerophages bear the CD4 antigen, the question was raised whether the immediate elTects observed may have been in part due to an influence on the mononuclear phagocyte system. Inimediately after MoAb infusions, a significant reduction of the absolute peripheral blood monocyte count down to 30%(P < 0·001) was noted within the firsl hour of injection. In contrast to strikingly elevated levels of soluble CD4 after treatment which was indicative of T cell lysis, soluble CD14 levels did not rise, but rather decreased from previously elevated levels. Before treatment, aetivation of the monocyte-macrophage system had been signified by elevated serum levels of IL-1, IL-6. CRP and neopterin as well as a marked in vitro production of IL-1. tumour necrosis faetor-alpha (TNF-α)and IL-6. Subsequent anti-CD4 treatment resulted in a rapid and significant reduction of monocyte-derived circulating cytokines and mediators concordant with a reduced capacity to produce IL-1. TNF-α, and IL-6 in those paiients who demonstrated clinical benefits. Therefore, studies of monocyte aetivation markers may be useful in identifying subsequent responders to anti-CD4 therapy.
T cell receptor haplotypes in families of patients with insulin-dependent diabetes mellitus
Authors: KELLY H , GARLEPP M J
Published: July 2010

The frequencies of Bgl 11 and BamH1 restriction fragment length polymorphisms (RFLP) of Cβ Vβ8, Vβ11 and Vβ7.2 have been defined in a healthy Australian population. Linkage disequilibrium between alleles of the T cell receptor (TCR) Vβ8 and Vβ11 gene segments has been confirmed. We have also confirmed the lack of linkage disequilibrium between either of these loci and alleles at Cβ or Vβ7.2. Using RFLPs at Vβ11 and Vβ8 loci TCR β haplotypes have been identified in five families in which the probands have insulin-dependent diabetes mellitus (IDDM). An extremely rare haplotype. marked by the higher molecular weight Bam H1 allele (H, H) at each of Vβ11 and Vβ8, was found in the DR4+ DR3− probands of two families (P= 0·004). In three families in which the probands had DR3. the more common TCR haplotype LH (Vβ11, Vβ8) was found. Taken together, these data confirm that linkage disequilibrium does exist in the TCR β locus, at least in some regions, and suggest that detailed analysis of the relationship between TCR Vβ haplotypes and HLA is warranted since these RFLPs may be markers for important allelic V gene sequence variations.
Identification and purification of a receptor on macrophages for the dengue virus-induced suppressor cytokine
Authors: MUKHERJEE R , CHATURVEDI P , CHATURVEDI U C
Published: July 2010

Dengue type 2 virus (DV)-induced suppressor cylokine (SF) binds to macrophages to transmit the suppressor signal to recruit the second subpopulation of suppressor T cells. The present study was undertaken to identify and purify the receptor for SF(SF-R) on macrophages. The binding of 125I-SF to macrophages was saturable and reversible. Scatchard analysis showed the presence of both high (54 000/cell) and low (178 × 106/cell) affinity receptor sites. The binding of 125I-SF to macrophages was inhibited by pretreatment of macrophages with anti-SF antiseruin but not by a heterologous antiserum. Normal mouse peritoneal macrophage membrane was solubilized with Triton-X-100 and the components separated by low pressure liquid chromatography (LPLC) to purify SF-R. The presence of SF binding moiety (SF-R) was screened at each step of purification. The purified SF-R resolved into two bands of 45–50 kD mol. wt on SDS-PAGE. 125I-SF + SF-R complex run on SDS-PAGE showed a single band at about 55–60 kD mol. wt by autoradiography. Anti-SF-R anti serum reacted with SF-R in a Western blot test; the reaction was abolished by pretreatment of the blots with proteinase K, but not by pretreatment with periodic acid. SF-R was composed of two polypeptide chains (α and β) which were obtained in pure form by high performance liquid chromatography (HPLC) of dithiothreitol- and iodoacetamide-treated SF-R. Only the β chain bound SF.
Effects of granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor on respiratory burst activity of neutrophiis in patients with myelodysplastic syndromes
Authors: OHSAKA A , KITAGAWA S , YUO A , MOTOYOSHI K , FURUSAWA S , MIURA Y , TAKAKU F , SAITO M
Published: July 2010

The superoxide (O2−-releasing capacity in response to N-fomiyl-methionyl-leucyl-phenylalanine (FMLP) and the priming effects of recombinant human granuloeyte colony-stimulating factor (rhG-CSF) and granulocyte-macrophage colony-stimulating faclor (rhGM-CSF) on FMLP-induced O2− release were investigated in neutrophils from 14 patients with myelodysplastic syndromes (MDS). The O2− -releasing capacity in MDS neutrophils varied from patient to patient. As compared with normal neutJ-ophils. the O2−-releasing capacity in MDS neutrophils was increased in 9/14 patients, nonnal in three patients and decreased in two patients. There was no close relationship between the O2−-releasing capacity and the peripheral blood neutrophil count or the plasma concentration of C-reactive protein. The priming of neutrophils by rhG-CSF was not observed in five patients, whereas rhGM-CSF primed neutrophils from all patients. The priming eflect of rhGM-CSF was consistently greater than that of rhG-CSF in each patient. The intravenous administration of rhG-CSF (300 μg/body) to two MDS patients showed an increase in the peripheral blood neutrophil count and enhancement of neutrophil O2 release. These findings demonstrate that the neutrophil O2−-releasing capacity in MDS varies from patient to patient and is not always impaired, and that rhGM-CSF is able to prime neutrophils which never respond to rhG-CSF.
Induction of mouse monocyte Ia expression by a mesangial cell-derived product
Authors: YAMASHITA W , LIU Y J , OOI B S
Published: July 2010

Previous studies in many laboratories have shown that macrophage hi expression is not constitutive but under regulation. We provide data which demonstrate that product(s) of mouse mesangial cell cultures Induce blood monocyte la expression as demonstrated by immunofluoresconce. This process is time-reialed and is also dependent on novel protein synthesis, being abrogated when themonocytes arepretreated with cycloheximidc. Preliminary characterization shows the mesangial cell product to be sensitive to healing at 100°C × 30 min, to be resistant to digestion by trypsin at a concentration of 4 × 10–6 M. and to have a molecular size of 10–100 kD as established by Amicon ultrafiltration. The substance is not interferon-gamma (IFN-γ) since cultured mesangial cells had no contaminating T cells, mesangial cell supernatant had no delectable levels of IFN-γ, and the Ia-inducing activity of the mesangial cell product was not abrogated by incubation of monoeytes with mesangial cell supernatant which had been immunoprecipitated with anti-IFN-γ. Similarly, experiments using anti-CSF-1 have excluded the possibility that the substance is CSF-1. The results of the study have relevance to the mechanisms by which inonocytes which take up residence in the glomerular mesangium acquire la positivity. and also provide a potentially novel pathway by which a tissue product may induce monocytes to express Ia.
Enhancement of IL-1, IL-2 production and IL-2 receptor generation in patients with acute rheumatic fever and active rheumatic heart disease; a prospective study
Authors: MORRIS K , MOHAN C , WAHI P L , ANAND I S , GANGULY N K
Published: July 2010

In a prospective study, patients with quiescent rheumatic heart disease (CRHD), streptococcal pharyngitis (SP) and healthy normal subjects produced comparable amounts of IL-1 and IL-2, but acute rheumatic fever (ARF) patients produced significantly elevated amounts of IL-1 and IL-2 at all intervals up to 48 weeks. In active rheumatic heart disease (ARHD), IL-1 activity returned to within normal range at 48 weeks, but IL-2 activity remained persistently elevated compared with CRHD, SP and healthy age- and sex-matched volunteers. CD4+ T lymphocytes were significantly increased in the peripheral blood of ARF and ARHD patients. The amount of IL-2 produced by ARF and ARHD patients correlated with the percentage of helper T lymphocytes (CD4+ cells) but not with the percentage of suppressor/cytotoxic T lymphocytes (CD8+ cells). Moreover, pre- and post-phytohaemagglutinin (PHA)-stimulated peripheral blood mononuclear cell (PBMC) cultures from ARF and ARHD patients contained higher proportions of IL-2R+ (CD25+) cells than those from patients with SP, CRHD and normal individuals, which persisted up to 48 weeks. The percentage of CD25+ cells in both types of PBMC cultures directly correlated with the percentage of CD4+ cells and not with CD8+ cells in active rheumatic patients only. These findings indicate that the immune response in ARF and ARHD patients is skewed to produce activated helper T cells that release IL-2 which drives the accumulation of more T helper cells. The result is an undamped helper T cell response in the peripheral blood of these patients.
Antigenic regions within the hepatitis C virus envelope 1 and non-structural proteins: identification of an IgG3-restricted recognition site within the envelope 1 protein
Authors: SALLBERG M , RUDÉN U , WAHREN B , MAGNIUS L O
Published: July 2010

Antibody binding to antigenic regions of hepatitis C virus (HCV) envelope 1 (El; residues 183–380), E2/non-structural (NS) 1 (residues 380–437), NS1 (residues 643–690), and NS4(1684–1751) proteins were assayed for 50 sera with antibodies to HCV (anti-HCV) and for 46 sera without anti-HCV. Thirty-four peptides. 18 residues long with an eight-amino acid overIap within each HCV region, were synthesized and tested with all 96 sera. Within the E region 183–380, the major binding site was located to residues 203–220, and was recognized by eight sera. Within the E2/NS1 region 380–437, the peptide covering residues 410–427 was recognized by two sera, and within the NSl region 643–690, peptides covering residues 663–690 were recognized by four sera. Within the NS4 region 1684–1751, 27 sera were reactive to one or more of the NS4 peptides, and 21 out of these were reactive with peptide 1694–1711. One part of the major binding site could be located to residues 1701–1704, with the sequence Leu-Tyr-Arg-Glu. The IgGl, IgG3 and IgG4 subcIasses were reactive with the five antigenic regions of HCV core, residues 1–18, 11–28, 21–38, 51–68 and 101–118. Reactivity to the major envelope site consisted almost exclusively of IgG3, and reactivity to the major site of NS4 consisted only of IgG1. Thus, a non-restricted IgG response to linear HCV-encoded binding sites was found to the core protein, whereas IgG subcIass-restricted linear binding sites were found within the El protein, and within the NS4 protein.
Modulation of human mesangial cell behaviour by extracellular matrix components—the possible role of interstitial type III collagen
Authors: SAITO K , SHIMIZU P , SATO T , OITE T
Published: July 2010

We have investigated the effects of various extracellular matrix (ECM) components on the behaviour of human mesangial cells (HMC) in u gel culture system using a modified MTT assay method. When cultured on u reconstituted basement membrane. Matrigel (M gel), HMC aggregated and formed isolated colonies initially, then extended an array of cell processes to form a dendritic network structure and proliferated very slowly as the culture period progressed. On type I colIagen gel (Cl gel), however. HMC developed elongated bipoIar shapes, migrated into the gel. and showed rapid cell growth. Next, separate ECM components, such as type III and IV colIagens. Iaminin. heparm and heparan sulphate, were incorporated into CI gel and HMC proliferation was assessed. Although attachment of HMC to each gel did not differ significantly. HMC proliferation was inhibited markedly on gels containing type III colIagen, heparin and heparan sulphate: type IV colIagen suppressed HMC proliferation slightly; and Iaminin had no significant effect. These data suggest that interstitial type I and III colIagcns. which arc often observed in diseased glomcruli. as well as the basement membrane components, may pIay important roles in the reguIation of HMC proliferation under pathophysiological conditions in vivo. We conclude that HMC behaviour is affected by the surrounding ECM constituents, which appear to function as a refined moduIator.
Rheumatoid arthritis: how well do the theories fit the evidence?
Authors: McCULLOCH J , LYDYARD P M , ROOK G A W
Published: July 2010

In this brief review, inspired partly by a symposium at the autumn meeting of the British Society for Immunology, 1992, varying hypotheses concerning the etiopathogenesis of rheumatoid arthritis (RA) are explored and tested against current evidence. Immunogenetic considerations, whilst of interest, have not aided our understanding of the development of this disease. The association with restricted HLA-DR β chain hypervariable sequences does not hold true with all cases of RA (but may be related to disease severity) and studies of T cell receptor (TCR) β chain usage fail to show consistent oligoclonality of infiltrating T cells in the synovial compartment. Etiologies based on triggering by bacteria are also considered: homologies between the ‘shared epitope’ sequences of HLA-DR 1 and DR4 β chains, Escherichia coli dnaJ and Proteus haemolysin do not indicate any feasible mechanisms for the development of RA, and cannot explain the many cases in which such DR sequences do not occur, though new data from man and animals enhance interest in the role of bowel flora. Finally, the striking parallels between slow bacterial infections and RA, in terms of immunogenetics, pathology, IgG glycosylation abnormalities and autoimmune manifestations, are put forward as circumstantial evidence that such bacterial infections may underly, or trigger, this serious disease.
Local production and localization of transforming growth factor-beta in tuberculous pleurisy
Authors: MAEDA J , UEKI N , OHKAWA T , IWAHASHI N , NAKANO T , HADA T , HIGASHINO K
Published: July 2010

Transforming growth factor-beta (TGF-β) is one of the cytokines which play an immunosuppressive role in an inflammatory process. To investigate the local production of TGF-β, we evaluated the levels of TGF-β in tuberculous pleural effusions (TBPE) and non-tuberculous benign pleural effusions (non-TBPE) by the growth inhibition assay with MvILu mink lung epithelial cells. The mean level of TGF-β in TBPE (46·1 ± 31·5 pM; mean ± s.d.) was higher than in non-TBPE (21·7 ± 12·3 pM) (P<0.05). Although the level of interferon-gamma (IFN-γ) in TBPE measured by ELISA was significantly higher than in non-TBPE. there was no significant difference in the levels of tumour necrosis factor-alpha (TNF-α) measured by ELISA between these two groups. Moreover, to elucidate localization of TGF-β in tuberculous pleurisy, immunohistochemical studies of pleura, using the rabbit polyclonal antibody Ab39 against latent TGF-β1 binding protein (LTBP) were performed. Results revealed that LTBP was localized in immature fibrotic areas where infiltrations of T lymphocytes and macrophages were absent. Importantly, the major sources of LTBP in these areas were thought to be mesothelial cells and fibroblasts. LTBP was not found in granulomas and mature fibrotic areas. Our data suggest that TGF-β in tuberculous pleurisy may play important roles for regression of granulomatous inflammation and pleural fibrosis for tissue repair.
Production and analysis of IgG monoclonal anti-DNA antibodies from systemic lupus erythematosus (SLE) patients
Authors: EHRENSTEIN M , LONGHURST C , ISENBERG D A
Published: July 2010

This study compares recently devised methods for producing IgG anti-DNA MoAbs from patients with SLE and analyses the antibodies generated from one patient at different phases of disease. Lymphocytes from SLE patients were transformed with Epstein-Barr virus (EBV) and/or fused with a heteromyeloma cell line, CB-F7. Direct fusion with CB-F7 resulted in the highest proportion of IgG-secreting lines, whereas EBV transformation resulted in a high percentage of IgM-secreting lines. Using direct fusion, five IgM anti-DNA antibody-secreting hybridomas were generated using lymphocytes from a patient with relatively inactive SLE. Six months later when the disease was active, only IgG anti-DNA antibodies were produced. The antigen-binding patterns of the MoAbs were analysed. Only one of the IgM anti-DNA antibodies reacted with dsDNA by ELISA and none by Crithidia immunofluorescence, whereas two of the IgG antibodies reacted with dsDNA by ELISA and Crithidia but did not bind to ssDNA. Only the two IgG high affinity anti-dsDNA antibodies bound to histones, and this was enhanced by added DNA, whereas three IgM antibodies bound to cardiolipin. This study supports the notion that MoAbs derived from a patient with SLE represent those found in the serum of SLE patients at different stages of disease activity. The binding to histones by the two IgG anti-dsDNA antibodies supports the recently expressed view that antibodies binding DNA/histone may be important in the pathogenesis of SLE.
Similar frequency of autoantibodies against 70-kD class heat-shock proteins in healthy subjects and systemic lupus erythematosus patients
Authors: KINDAS-MÜGGE I , STEINER G , SMOLEN J S
Published: July 2010

Stress or heat-shock proteins may be involved in the initiation and perpetuation of autoimmune diseases. In order to investigate a possible role of autoantibodies against the 70-kD family of heat-shock proteins in systemic lupus erythematosus (SLE), sera of SLE patients and healthy subjects were tested for the presence of IgG and IgM antibodies to 70-kD class proteins. These proteins were purified by affinity chromatography on ATP-agarose and used in Western blotting studies. The data obtained revealed that antibodies to the 72-kD and the 73-kD heat-shock proteins occurred with similar frequencies both in healthy subjects and SLE patients. Thus, approximately 20% of the sera in each group contained IgG antibodies, and IgM antibodies were detected in about 30% of the sera tested. Moreover, in SLE patients no association between the occurrence and litre of these antibodies and disease activity was found. These data suggest that antibodies to the 70-kD class heat-shock proteins are naturally occurring and argue therefore against an involvement of these antibodies in the pathogenesis of SLE.
Differential induction of helper T cell subsets during blood-stagePlasmodium chabaudiAS infection in resistant and susceptible mice
Authors: STEVENSON M M , TAM M-F
Published: July 2010

The induction of T helper cell subsets during the course of non-lethal or lethal blood-stage Plasmodium chabaudi AS infection was investigated using inbred strains of mice which differ in the level of resistance to this intraerythrocytic parasite. Resistant C57B1/6 mice experience a non-lethal course of infection characterized by moderate levels of both parasitaemia and anaemia and resolution of primary acute infection by 4 weeks, while susceptible A/J mice experience lethal infection with fulminant parasitaemia and severe anaemia. T helper subset function was assessed during infection by determining the kinetics of spleen cell production in vitro of the Th1-derived cytokine, interferon-gamma (IFN-γ), and of the Th2-dcrived cytokine, IL-5, using sandwich ELISAs. Spleen cells from resistant C57B1/6 mice were found to produce high levels of IFN-γ within 1 week of infection in response to both the mitogen concanavalin A (Con A) and malaria antigen. Furthermore, CD4+ T cells were found to be the source of I FN-γ while both CD4+ and CD8+ T cells were found to produce IL-5. Decreased IFN-γ production after day 10 was concomitant with significant production of IL-5 between 2 and 3 weeks post infection. In contrast, spleen cells from susceptible A/J mice produced high levels of IL-5 within the first week of infection. In addition, these animals were found to have high serum levels of IL-5. These results, thus, confirm previous observations that resolution of primary blood-stage P. chabaudi infection occurs by sequential activation of Th1 CD4+ T cells followed by activation of the Th2 subset, and in addition, suggest that induction of a strong Th2 response early in infection may lead to a severe and lethal course of malaria.
Phenotypic characterization of CD8+ T cell populations in HIV disease and in anti-HIV immunity
Authors: WATRET K C , WHITELAW J A , FROEBEL K S , BIRD A G
Published: July 2010

The CD8+ T cell population is believed to play an important role in the control of viral infection, both for suppression of viral replication and for cytotoxic activity against viral infected cells. Elevated numbers of CD8+ T cells have been demonstrated in HIV infection, and CD8+ cytotoxic T cell (CTL) activity is associated with the early, asymptomatic stage of disease. We investigated the phenotypic characteristics of the CD8 population, in whole blood, in HIV disease and determined the predominant CD8+ subpopulation involved in anti-HIV CTL activity. We found that CD8+ T cells co-expressing markers of activation (HLA-DR), memory (CD45RO, CD29), and cytotoxic activity (S6F1) were significantly elevated in the early stages of disease, while the numbers of naive (CD45RA) cells remained unchanged. Progression to AIDS resulted in an overall loss of absolute CD8+ T cells, though the percentages of CD8+ HLA-DR+ and CD8+ S6F1+ remained elevated. In contrast to patients in the late stages of disease, anti-HIVgag CTL activity, following in vitro stimulation, was present in most HIV+ asymptomatic subjects and was associated with an expansion of CD8+ HLA-DR+ and CD8+ CD45RO+ cells. The absence of CTL activity was associated with a reduced ability of these populations to expand in vitro and with a significant loss of peripheral CD4+ T cells, independent of clinical stage. We suggest that CD8+ expressing HLA-DR+ CD45RO+ and S6F1+ play an important role in anti-HIV cytotoxicity.
Increased expression of interferon-gamma in hyperplastic lymph nodes from HIV-infected patients
Authors: BOYLE M J , BERGER M F , TSCHUCHNIGG M , VALENTINE J E , KENNEDY B G , DIVJAK M , COOPER D A , TURNER J J , PENNY R , SEWELL W A
Published: July 2010

Polyclonal B cell activation is characteristic of HIV infection and occurs in the presence of severe CD4+ lymphocyte depletion. In contrast, CD4+ lymphocytes are the dominant T cell in the reactive lymphoid tissues of patients not infected with HIV. In this study, lymph node biopsies from eight HIV-infectcd patients with persistent generalized lymphadenopathy syndrome (PGL) were assessed for IL-lβ, IL-2, IL-4, IL-6, IL-10, interferon-gamma (IFN-γ) and tumour necrosis factor-beta (TNF-β) gene expression using the polymerase chain reaction (PCR). The cytokine gene expression of two cases of reactive adenopathy in patients not infected with HIV was assessed for comparison. IFN-γ was expressed much more strongly in the PGL samples than in control reactive lymphoid tissues, whereas the other cytokines were expressed to a similar extent in both types of tissues. IFN-γ may have an important role in maintaining the adenopalhy of HIV-infected patients. Expression of cytokines such as IL-2, IL-4 and IL-10 in HIV nodes may be adequate to allow the recruitment of naive B cells to the reactive process.
Vitronectin-mediated inhibition of complement: evidence for different binding sites for C5b-7 and C9
Authors: MILIS L , MORRIS C A , SHEEHAN M C , CHARLESWORTH J A , PUSSELL B A
Published: July 2010

In the activated complement system, vitronectin (complement S-protein) occupies the metaslable membrane binding site of the nascent precursor complex C5b-7, so that the newly formed SC5b-7 is unable to insert into cell membranes. Some evidence also indicates that vilronectin limits on-going membrane-associated pore formation by inhibiting C9 polymerization. It has been assumed that these two stages of terminal complement complex (TCC) inhibition take place through charge interactions between the heparin-binding region of vitronectin and homologous cysteine-rich sequences of the late complement proteins C6, C7, C8 and C9. We examined SC5b-7 formation and inhibition of C9 binding in the TCC using separate haemolytic assays. The mode of action of vitronectin in these assays was compared with two 15mer peptides which span residues 348–379 of the heparin-binding region, and a heparin-attinity polypeptide. protamine sulphate. The results showed that vitronectin acts predominantly through SC5b-7 production with a lesser effect on the inhibition of C9 lytic pore formation. In contrast, protamine sulphate did not prevent C5b-7 membrane attachment, but was a potent inhibitor of C9-mediated lysis. The peptides did not inhibit C5b-7 membrane insertion and only one affected C9 binding. These data suggest that the two stages of TCC inhibition involve separate binding sites on the vitronectin molecule. The site for association with nascent C5b-7 is unknown, whereas inhibition of C9 binding and pore formation lakes place through the heparin-binding region.
The ABA-1 allergen of the nematodeAscaris suum: epitope stability, mass spectrometry, and N-terminal sequence comparison with its homologue inToxocara canis
Authors: CHRISTIE J F , DUNBAR B , KENNEDY M W
Published: July 2010

ABA-1 is a major allergen of nematode parasites of the genus Ascaris which includes the large roundworms of humans and pigs, A. lumbricoides and A. suum, respectively. The allergen was purified from A. suum by immunoaffinity chromatography for immunochemical examination. The IgE antibody repertoire is under MHC control in infected rodents and the IgE-binding epitopes were robust to treatment with heat or periodate, and electroblotting on nitrocellulose. This implies that the IgE epitopes comprise primary peptide sequence or an unusually stable secondary or tertiary structure. The molecular mass of ABA-1 is controversial, but mass spectrometry analysis indicated that there were five components of similar size, with the major species being 14 643·2 ± 1·4D. Finally, N-terminal sequence analysis of ABA-1 and TBA-1 (the homologue in the canine nematode infective to humans, Toxocara canis) revealed a high degree of similarity, and we have previous evidence that ABA-1 homologues are widespread amongst ascaridid parasites of humans. ABA-1 and its homologues might, therefore, be important to the immunopathology of many infections with nematode parasites, upon which the genetic constitution of the hosts will also have a bearing.
Abnormalities of the IgA immune system in members of unrelated pedigrees from patients with IgA nephropathy
Authors: SCHENA F P , SCIVITTARO V , RANIERI E , SINICO R , BENUZZI S , DI CILLO M , AVENTAGGIATO L
Published: July 2010

In the last few years many investigators have reported the recurrence of primary IgA nephropathy (IgAN) or the presence of persistent microhaematuria and/or proteinuria in family members of patients with IgAN. Our study was undertaken to investigate the relevance of abnormalities in the regulation of the IgA and IgM immune system in microhacmaturic and asymptomatic family members of IgAN patients. Fifty-four out of 120 members of nine unrelated pedigrees were examined by urinalysis; polymeric IgA (pIgA). IgA rheumatoid factor (IgARF), IgAI-IgG immune complexes (IgAl-IgG IC) and IgAl-IgM IC, and other immunoglobulins were measured in serum samples. Moreover, we studied the production of immunoglobulins. pIgA and IgARF by peripheral blood mononuclear cells (PBMC) in basal conditions and after pokeweed mitogen (PWM) stimulation. Our data demonstrate that persistent mierohaematuria was present in 24% of relatives. High serum levels of IgA. mainly pIgA and IgARF, IgAI-IgG IC and IgAl-IgM IC occurred in 66% of relatives. Abnormal spontaneous production of IgA by PBMC and after PWM stimulation was present in 64% of family members. Interestingly, high scrum levels of IgM and abnormal production of this immunoglobulin by PBMC were observed in relatives. However, the immunological abnormalities did not correlate in any way with the presence of urinary abnormalities such as microhaematuria. which was most likely determined by an underlying glomerular alteration.
Augmentation by transferrin of IL-2-inducible killer activity and perform production of human CD8+ T cells
Authors: NAKAMURA A , SONE S , NABIOULLIN R , SUGIHARA K , MUNEKATA M , NISHIOKA Y , NII A , OGURA T
Published: July 2010

The effects of human transferrin (Tf) on lymphokine (IL-2)-activated killer (LAK) induction from blood lymphocytes of healthy donors was examined. LAK cells were induced by 6-day incubation in medium with recombinant human IL-2 of lymphocytes, and their cytotoxic activity was assessed by measuring 51Cr release from NK-resistant Daudi cells. Tf alone did not induce any LAK activity, but in combination with IL-2, it augmented LAK induction dose- and time-dependently. This augmenting effect was completely abolished by pretreatmenl with anti-Tf antiserum. Tf augmented the proliferative response of lymphocytes to IL-2 and their expressions of receptors for IL-2 and Tf. CD8+ T cells were isolated from purified blood lymphocytes using antibody-bound magnetic beads. Addition of Tf to cultures of CDS+ cells resulted in significant augmentation of killer cell induction and perforin (PFP) production after 4 days stimulation with IL-2. These results indicate that Tf is important in generation of IL-2-inducible killer properties and PFP activity of human CD8+ T cells.
The functional affinities of antibodies of different IgG subclasses to dietary antigens in mothers and their babies
Authors: DEVEY M E , BECKMAN S , KEMENY D M
Published: July 2010

The quantity and functional affinities of IgG1 and IgG4 antibodies to the dietary antigens casein and ovalbumin were measured in unselccted mothers and their 1-year-old infants. In these infants, the titre of IgG antibodies to both antigens was highest in the IgG1 subclass, while in their mothers the litre of IgG1 and IgG4 antibodies to these foods was similar. High affinity IgG4 responses to both casein and ovalbumin were frequently found in mothers, whilst IgG1 responses, particularly to ovalbumin. were of 1ow functional affinity. By contrast, in the 1-year-old infants, the functional affinity of IgG1 antibody to ovalbumin was substantially higher than in their mothers (Student's paired t-test, P < 0·001). indicating that higher affinity IgG1 antibody was produced on first exposure to ovalbumin rather than following chronic exposure. The effect of antigenic load on affinity maturation was further investigated by comparing the affinity of IgG1 antibody to casein in bottle, mixed and breast fed infants. Bottle fed infants had significantly higher-affinity IgG1 antibodies to casein compared with breast or mixed fed infants (Student's unpaired t-test. P < 0·01 and 0·02), suggesting that antigen exposure via the gut was able to drive the affinity maturation process. In studying the immune response it is clear that account must be taken of the affinity as well as of the litre of the antibody produced.
Regulation of resistance against adjuvant arthritis in the Fisher rat
Authors: van de LANGERIJT A G M , van LENT P L E M , HERMUS A R M M , van de PUTTE L B A , van den BERG W B
Published: July 2010

Inbred female Lewis (LEW/N) rats develop a severe chronic arthritis in the adjuvant arthritis (AA) model, histocompatible Fisher(F344/N) rats are resistant and germ-free Fishers (GF F344) are again susceptible. In this study we show that the F344 rat can become susceptible to AA, using Mycobavterium tuberculosis (M.tb.) in the powerful adjuvant paraffin oil, instead of mineral oil (Freund's incomplete adjuvant (FIA)). This indicates that the F344 rat does nol lack T effector cells. To examine further mechanisms responsible for suppression, we determined the level of plasma eorticosterone in response to IL-1α in Lewis. F344 and GF F344 rats. IL-lα induced only low amounts of corticosterone in Lewis rats, but high amounts in both F344and GF F344 rats. The GF F344 rats are susceptible lo AA. but the severity of the disease is redueed compared with Lewis rats. This indicates that corticosterone may be an important mechanism lo suppress disease development, but not the only mechanism. In addition we investigated whether T suppressor cells play a role in the resistance of the F344 strain. This was performed by pretreating the animals with the immunomodu-laling drugs cyelophosphamide (Cy) and cyclosporin A (CsA). We were unable to make the F344 rat susceptible to AA, indicating that active suppression does nol play a role in the induction phase of arthritis. This finding is confirmed in adoptive transfer experiments of AA from Lewis to F344 rats. Our dala suggest the lack of a strong pre-existing suppression in the F344 rats, and indicate that suppression is generated upon bacterial challenge. Whether suppression is overruled probably depends on the power of adjuvants used and potential control by corticosteroids.
Immunohistochemical demonstration of membrane cofactor protein (MCP) of complement in normal and diseased kidney tissues
Authors: ENDOH M , YAMASHINA M , OHI H , FUNAHASHI K , IKUNO T , YASUGI T , ATKINSON J P , OKADA H
Published: July 2010

The immunohistochemically stained membrane cofactor protein of complement (MCP/CD46), one of the complement regulatory proteins, was up-regulated in some diseased kidney tissues. MCP in diseased kidneys was strongly concentrated along the glomerular capillary walls as well as In the mesangial regions, while MCP in normal kidneys was weakly detected in all glomerular structural cells and in the epithelial cells of tubules. Since the enhanced staining was noted in those areas where depositions of C3b/C3c occurred, ongoing complement reaction might be responsible for the up regulation of MCP expression. MCP expression may be up-regulated by complement fragments generated during complemenl activation in glomerulonephritis. Furthermore, anti-MCP staining was stronger in intensity in patients with moderate to massive proteinuria, indicating that up regulation of MCP expression could be directly correlated to the kidney damage.
Population study of T cell receptor Vβ gene usage in peripheral blood lymphocytes: differences in ethnic groups
Authors: GEURSEN A , SKINNER M A , TOWNSEND L A , PERKO L K , FARMILOE S J , PEAKE J S , SIMPSON I J , FRASER J D , TAN P L J
Published: July 2010

The T ceil receptor (TCR) Vβ repertoire in peripheral biood lymphocytes (PBL) of a largc number of healthy individuals was analysed by quantifying Vβ-specific mRNA using the method of anchored muitiprimer DNA ampiification and a reverse dot blot assay. Among 16 Vβ gene families examined, particular Vβ genes were noted to be unequally expressed in the PBL of 70 healthy donors. The frequently used genes belong to the Vβ 4, 5, 6, 8 and 13 (12) families, while Vβ 1, 9 and 15 were the least frequently used gene families. This bias in gene usage was observed in all individuals. Marked deviation from the mean percentage usage was noted for some Vβ genes in individuals when their PBL were examined serially. but the common pattern of biased usage was not grossly distorted. When the TCR repertoire of different ethnic groups was examined, a lower mean frequency of Vβ 3·2 was seen in the repertoire of 19 Caucasians compared with 25 age-matched Samoans (P< 0·003). Conversely, the expression of Vβ 5·1 and of Vβ 5·3 was higher in Caucasians than in 5 age-matchcd Polynesians (Maoris and Samoans, P< 0·003). Considering the 20% co-efficicnt of variation in the estimate of Vβ gene usage, our data from 70 unrelated individuals suggest that in PBL, individual variations in the TCR repertoire were superimposed upon a common biased usage of Vβ genes in the general population.
Effects of activin A on IgE synthesis and cytokine production by human peripheral mononuclear cells
Authors: YAMASHITA N , NAKAJIMA T , TAKAHASHI H , KANEOKA H , MIZUSHIMA Y , SAKANE T
Published: July 2010

Activin A not only stimulates the synthesis and release of pituitary follicle-stimulating hormone, but exerts various effects on haematopoietic cells. embryos, and fibroblasts. In the present study we have examined effects of activin A on lgE synthesis and cytokine production by peripheral blood mononuclear cells (PBMC) in normal humans. When PBMC were cultured in the presence of IL -4, activin A significantiy augmented IgE production induced by IL-4. Activin A did not affect, however, IgE production from highly purified B cells when they were stimulated with anti-CD40 MoAb and IL-4. The fact that in the latter condition IgE synthesis was T cells- and monocyte-indcpcndent indicated that activin A does not directly influence B cells for IgE synthesis. Rather, production as well as gene expression of IL-6, which is known to enhance IgE synthesis by purified monoeytes, was induced by activin A alone. In addition, activin A induced other monokines such as IL-l and tumour necrosis factor (TNF)-α from monocytes. In contrast, activin A neither induced nor augmented the production of TNF-β or interferon-gamma (IFN-γ), both of which are known to be exclusively generated by T cells. These data indicate that activin A plays a certain role in physiological functions for monocytes in normal humans.
Trypanosoma cruziinfection enhances polyreactive antibody response in an acute case of human Chagas' disease
Authors: GRAUERT M R , HOUDAYER M , HONTEBEYRIE-JOSKOWCIZ M
Published: July 2010

The kinetics of antibody response in an acute case of human Chagas' disease was investigated. Hypergammaglubulinaemia appeared at day 17 of infection, and persisted after 66 days of infection, at which time parasitaemia became undetectable. Titration of immunoglobulins showed that the three principal isotypes were involved in the response, emphasizing polyclonal B cell activation. Total IgA was detected before total IgM, and the killer before total IgG. High titres of autoantibodies were found among IgM and IgG subclasses. IgA was also the first isotype to be detected among specific anti-Trypanosoma cruzi antibodies. However, the maximal parasite antibody response was attained after 30 days of infection for all isotypes. With regard to possible cross-reactivity between molecules of host and parasite, adsorption experiments on T. cruzi-specific immunosorbent were designed. Specific antibodies, present in the eluates, also recognized natural antigens, especially laminin. In order to characterize the α-galactose epitope of laminin, adsorption experiments on sheep erythrocytes were performed, and revealed the possible presence of another epitope on the glycoprotein. Our results indicate that in the case of Chagas' disease investigated here, polyclonal activation occurred; moreover, they suggest that molecular mimicry may play a role by increasing autoantibodies, probably via a parasite-driven mechanism.
Increased IFN-α-induced sensitivity but reduced reactivity of 2′,5′-oligoadenylate synthetase (2,5AS) in trisomy 21 blood lymphocytes
Authors: GERDES A-M , HØRDER M , BONNEVIE-NIELSEN V
Published: July 2010

2,5 AS is induced by interferon-alpha, -beta (IFN-α, -β) and polymerizes adenosine triphosphate (ATP) into pppA (2′ p5′ A) oligomers, leading to inhibition of protein synthesis and virus growth. The gene coding for the IFN-α, -β receptor is localized lo chromosome 21. In trisomy 21 cells from patients with Down's syndrome a 50% increase in gene dosage is present for the genes on this chromosome. Corresponding increase of gene products has been shown for several of these genes, e.g. The IFN-α, -β receptor molecules. Here, we show a direct proportionality between the number of IFN-α, -β receptors and both the basal activity of 2,5AS and the sensitivity of this enzyme for IFN-α, -β. The relative increase of 2,5AS activity (the reactivity) becomes reduced consistent with the increased basal 2,5AS activity in trisomy 21 cells compared with control cells.
Human colonic intraepithelial lymphocytes from patients with inflammatory bowel disease fail to down-regulate proliferative responses of primed allogeneic peripheral blood mononuclear cells after rechallenge with antigens
Authors: DALTON H R , DIPAOLO M C , SACHDEV G K , CROTTY B , HOANG P , JEWELL D P
Published: July 2010

Human colonic intraepithelial lymphocytes from control subjects down-regulate the proliferative responses of primed allogeneic peripheral blood mononuclear cells on rechallenge with antigens or phytohaemagglutinin (PHA). In contrast, human colonic intraepithelial lymphocytes from patients with inflammatory bowel disease fail to down-regulate the proliferative responses of primed allogeneic peripheral blood mononuclear cells on rechallenge with antigens. These findings may be important in the development and maintenance of the mucosal immunological activation of inflammatory bowel disease.
Expression of types I, III and IV collagen genes in fibrotic skin and nerve lesions of toxic oil syndrome patients
Authors: GOMEZ-REINO J J , SANDBERG M , CARREIRA P E , VUORIO E
Published: July 2010

We have studied the skin and nerve fibrosis in toxic oil syndrome by in situ hybridization using specific cDNA probes for types I, III, and IV collagens, Fibroblasts with high levels of type I and III collagen mRNA were observed in biopsies from fibrotic skin areas. Similarly, type IV collagen mRNA was abundant in cells within the fibrotic process of the nerves. These results suggest that the excessive accumulation of collagen in toxic oil syndrome results from transcriptional activation of collagen genes in a subpopulation of fibroblasts.
Partial prevention of active Heymann nephritis by 1α, 25 dihydroxyvitamin D3
Authors: BRANISTEANU D D , LEENAERTS P , van DAMME B , BOUILLON R
Published: July 2010

The hormone 1α, 25 dihydroxyvitamin D3 (1,25(OH)2DO3 has potent immunosuppressivc effects in vitro. Recent publications also described a protective effect of the hormone in various animal models of immune-mediated diseases. To test its in vitro activity we induced active Heymann nephritis in Lewis rats that were either untreated or treated with 1,25(OH)2D3 or its synthetic 20-epi analogue, KH1060. Treatment with cyclosporine A (CsA) was used as an immunosuppressive control. In this nephrotic model the administration of 1,25(OH)2D3 (0.5 μg/kg body weight) given on alternate days during the first 13 days after active immunization significantly reduced the proteinuria as measured by weeks 7–9. This reduction was comparable to the reduction observed in rats treated with CsA (20 mg/kg) on alternate days. A second series of experiments with 1,25(OH)2D3 confirmed these findings. The level of autoantibodies was found to be significantly suppressed during the treatment time in the CsA (20 mg/kg) group, whereas the limit of significance (P=0.06) was reached in the 1,25(OH)2D3 groups that developed less proteinuria. The administration of 1,25(OH)2D3 transiently increased the mean serum calcium Concentration with 2.5 mg/dl above the pretreatmcnt values, and the urinary calcium excretion by a factor of 3–5 during the short treatment time. Treatment with the analogue KH 1060 did not reduce the proteinuria significantly. Our experiments add evidence to the hypothesis that 1,25(OH)2D3 in pharmacological doses has immunosupprcssive potency.