bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cytomegalovirus hepatitis: characterization of the inflammatory infiltrate in resistant and susceptible mice
Authors: OLVER S D , PRICE P , SHELLAM G R 
Published: July 2010

SUMMARYMice susceptible and resistant to murine cytomegalovirus (MCMV) were infected with this virus and livers were harvested after 2–231 days. Cryostat sections were stained to visualize cells bearing CD4, CDS8 or Mac-1 antigens. Mac-1+ cells were prevalent in inflammatory foci after 2 days. These cells persisted in susceptible BALB/c and A/J mice, but disappeared from livers of resistant C57BI/6 and CBA/CaH mice by day 28. T cell inflammation peaked on days 7–11. This declined by day 56 in C57BI/6 and CBA/CaH mice, but persisted in BALB/c and A/J mice for at least 231 days. Persistent CD8+ cells were dispersed throughout the parenchyma. More CD8+ cells were observed 7–14 days after infection in the livers of bg/bg (natural killer (NK) cell-deficient) C57B1/6 and CBA mice, and in C57BI/6 mice depleted of NK 1.1 cells by MoAb. Thus, mice of strains susceptible to MCMV exhibit hepatitis characterized by persistence of dispersed CD8+ cells. This phenomenon may be limited by NK ceils in resistant strains.
Intranasal inoculation ofMycoplasma pulmonisin mice with severe combined immunodeficiency (SCID) causes a wasting disease with grave arthritis
Authors: EVENGÅRD B , SANDSTEDT K , BÖLSKE G , FEINSTEIN R , RIESENEELT-OURN I , SMITH C I E 
Published: July 2010

SUMMARYMycoplasma pulmonis or Myc. pneumoniae were inoculated intranasally to C.B-17 scid/scid mice (severe combined immunodeficient (SCID) mice). Immunocompetent C.B-17 mice were inoculated as controls. During the observation period of 5 weeks the mice were killed and necropsied. Mycoplasma pulmonis was recovered from all of the inoculated mice, and dissemination to various tissues increased with time. SCID mice, unlike immunocompetent mice, did not show lung lesions but exhibited severe inflammatory changes of the joints. Mycoplasma pulmonis, however, was isolated both from the lungs and the articular lesions. In addition, SCID mice infected for more than 3 weeks suffered from a pronounced loss of weight and emaciation. In the experiment with Myc. pneumoniae the agent could be reisolated, but lesions were not found in any of the infected mice. Mycoplasma pulmonis infection in SCID mice may be useful as a model of arthritis in immunodeficient humans.
Sequential autoantigenic determinants of the small nuclear ribonucleoprotein Sm D shared by human lupus autoantibodies and MRLIpr/lprantibodies
Authors: JAMES J A , MAMULA M J , HARLEY J B 
Published: July 2010

SUMMARYAutoantibodies directed against the Sm proteins of the spliceosome complex are found in approximately 25% of systemic lupus erythematosus (SLE) patient sera. To determine which regions of the Sm D polypeptide are involved in the lupus autoimmune response, binding to overlapping octapeptides of Sm D has been evaluated with sera from nine Sm D-positive patients, six patients with other autoimmune serology, and five normal human sera. Lupus patient sera which are Sm precipitin-positive bind various combinations of five regions of the peptide. The major antigenie region. Epitope 5 (REAVA(GR)10 GGPRR), is bound by eight of nine Sm precipitin-positive sera tested. This region of Sm D shows significant sequence homology with Epstein-Barr nuclear antigen-1. To determine the fine specificity of the murine Sm response, four unique Sm D MoAbs derived from MRL lpr/lpr mice and three adult anti-Sm-positive MRL lpr/lpr mouse sera have been analysed. Two of these monoclonals, KSm4and Y12, as well as the MRL lpr/lpr sera tested, show binding with Epitope 5. Another of these monoclonals, KSm 2, binds octapeptides 84–91, DVEPKVKSKKREAVAG, which corresponds to Epitope 4 of this study. Antibodies from SLE patients with autoimmune serology other than anti-Sm bind the carboxyl glycine-arginine repeat (GR)10 peptides of Sm D. However, none of the antibodies tested from patients who do not have lupus and who have different autoimmune serology binds any of the Sm D octapeptides. Normal controls did not significantly bind any of the Sm D octapeptides. These results describe two major regions of shared antigenicity of Sm D between sera from SLE patients and MRL lpr/lpr mice, thereby establishing a basis for the cross-species similarity of autoimmunity to the Sm autoantigen in SLE.
Enhanced synthesis of cytokines by peripheral blood monocytes cultured in the presence of autoantibodies against U1-ribonucleoprotein and/or negatively charged molecules: implication in the pathogenesis of pulmonary hypertension in mixed connective tissue disease (MCTD)
Authors: OKAWA-TAKATSUJI M , AOTSUKA S , UWATOKO S , SUMIYA M , YOKOHARI R 
Published: July 2010

SUMMARY
An attempt was made to determine whether addition of purified autoantibodies against Ulribonucleoprotein (RNP) and negatively charged molecules (cardiolipin and double-stranded (ds) DNA) to cultures of peripheral blood monoeytes could enhance the synthesis of eytokines in patients with MCTD and normal healthy volunteers. It was found that: (i) at the baseline, levels of cytokines such as IL-1α, IL-β and IL-6 extracellularly released by or associated with monocytes were significantly higher in MCTD patients than in normal subjects; (ii) addition of antibodies against U1-RNP to cultures of MCTD monocytes resulted in a significant overall increase of the released and cell-associated IL-1α, IL-β and IL-6. On the other hand, addition of antibodies against cardiolipin or dsDNA to cultures of MCTD monocytes resulted in a significant increase of released and/or cell-associated IL-1α and IL-1β; (iii) addition of these autoantibodies to cultures of normal monocytes resulted in a significant overall increase of released and cell-associated IL-1α, IL-1β and IL-6. The extent of enhancement of cytokines released by or associated with monocytes was greater in normal subjects than in MCTD patients; (iv) a F(ab′)2 preparation of autoantibodies against U1-RNP also enhanced the level of released and cell-associated IL-1α. Our findings that both autoantibodies against U1-RNP and negatively charged molecules were able to enhance the synthesis of cytokines by monocytes suggest that these autoantibodies might cause derangement of endothelial cells and lead to proliferative vaseulopathy, which is a characteristic of pulmonary hypertension in MCTD.
Tryptic peptides of human thyroglobulin: II. Immunoreactivity with sera from patients with thyroid diseases
Authors: SABOORI A M , CATUREGLI P , ROSE N R , MARIOTTI S , PINCHERA A , BUREK C L 
Published: July 2010

SUMMARYTryptic peptides of human thyroglobulin (Tg) were analysed by Western immunoblot for their reactivity to circulating autoantibodies from patients with Hashimoto's thyroiditis (HT), Graves’ disease (GD) and thyroid carcinoma, and from normal human controls. Low molecular weight peptides were released after 4h incubation of Tg with trypsin. The sera of thyroid disease patients reacted with several peptides, but predominantly bound three peptides with apparent molecular weights (MWap) of 25 kD, 20 kD, and 15kD; the sera of normal individuals did not bind these fragments of Tg. The pattern of Tryptic peptides recognized by the majority of sera from GD patients differed from that recognized by sera from most patients with HT. Autoantibodies from both groups of patients recognized a 15-kD peptide with a high frequency, but the sera from 26/43 (60%) GD patients also recognized a peptide with MWap of 25 kD, whereas the sera from 22/35 (63%) of HT patients recognized a 20-kD peptide. A few sera from patients with thyroid carcinoma reacted with peptides with MWap of 15 and 20-kD, and none bound the 25-kD peptide. The immunoreactivity of autoantibodies in HT sera to the 20-kD peptide paralleled the competitive inhibition of the MoAb 137CI by these sera. In addition, MoAb 137CI and Hashimoto's sera showed the same Western immunoblot-binding pattern to Tg tryptic peptides, suggesting that a Hashimoto-associated epitope and the 137Cl-binding site are found on the same peptide. These findings suggest that distinct peptides are recognized by Tg autoantibodies from patients with different thyroid diseases.
Characterization of naturally occurring autoantibodies against tumour necrosis factor-alpha (TNF-α):in vitrofunction and precise epitope mapping by phage epitope library
Authors: SIOUD M , DYBWAD A , JESPERSEN L , SULEYMAN S , NATVIG J B , FØRRE Ø 
Published: July 2010

SUMMARYNaturally occurring autoantibodies against cytokines exist in the sera of patients with autoimmune diseases as well as in the sera of normal individuals. We report here that affinity-purified autoantibodies against human TNF-α from one rheumatoid arthritis (RA) patient inhibited the cytotoxic effect of TNF-α on the mouse fibrosarcoma cell line WEHI 164, by 50%. In an attempt lo predict the autoantibodies” recognition site on TNF-α protein we screened a random nanopeptide phage library with the affinity-purified TNF-α autoantibodies. Among 63 random selected clones, 46 clones carried the sequence ASSLLASSP, NSSPYLNTK or PQSPGSSFP. Frequency analysis of the relative occurrence of the 20 amino acids in the nanopeptides displayed by 50 random bacteriophages picked before selection and 63 after selection to bind to TNF-α autoantibodies indicated that proline (P &lt; 0·0003) and serine (P &lt; 0·04) are involved in the binding of the autoantibodies to the phages. Furthermore, we demonstrated that three synthetic peptides ASSLLASSP, NSSPYLNTK and PPLFCPVTDE displayed by the selected phages reduced the binding of the autoantibodies to TNF-α protein by 50%. Interestingly, the sera of mice (BALB/c) immunized with phages displaying ASSLLASSP and NSSPYLNTK peptide showed an anti-TNF-d response as detected by ELISA. This response was not found in mice immunized with the wild type phage. Thus, the recombinant phages selected from the phage libraries could be used as carrier for immunization, and therefore as a tool for vaccine development. This work sets the stage for experiments designed to isolate ligands for protective antibodies.
Flow cytometry analysis of OKT4 epitope deficiency in South African black children
Authors: HUGHES E J , GODDARD F A , BOUIC P , BEATTY D W 
Published: July 2010

SUMMARYA case of OKT4 epitope deficiency referred for investigation with suspected immunodeficiency is described. Flow cytometry analysis of OKT4 epitope deficiency in a study group of healthy black children showed different manifestations of the lack of OKT4 epitope; a complete lack of OKT4+CD4+ peripheral blood lymphocytes (PBL) with normal numbers of OKT4A+ and Leu-3a+CD4+ PBL, decreased percentage OKT4+CD4+ compared with OKT4A+ and Leu-3a+CD4+ PBL, decreased fluorescent staining intensity with OKT4 and a biphasic OKT4 staining pattern associated with a reduced OKT4/Leu-3a ratio. The percentage and fluorescent intensity of OKT4+CD4+ PBL in the study group were significantly lower (P &lt; 00001) than Leu-3ab+CD4+ and OKT4A+CD4+ PBL. There is thus considerable risk of under-estimating the number of CD4+ cells in black South Africans if the OKT4 MoAb is used.
Immunopathological features of palatine tonsil characteristic of IgA nephropathy: IgA1 localization in follicular dendritic cells
Authors: KUSAKARI C , NOSE M , TAKASAKA T , YUASA R , KATO M , MIYAZONOU K , FUJITAIJ T , KYOGOKU M 
Published: July 2010

SUMMARYIgA nephropathy (IgAN) is generally thought to be mediated by the glomerular deposition of circulating immune complexes containing IgA as the major antibody component. Upper respiratory infections and tonsillitis often precede IgAN. and in some cases tonsillectomy is affective for the (treatment of IgAN. Thus, the tonsil seems to be a unique organ causing initial and/or progressive events to generate nephritogenic immune complexes in IgAN. in this study we focused on the analysis of immunopathological features of the palatine tonsil characteristic of IgAN patients by using an immunohistochemical technique. The IgAl subclass was demonstrated in follicular dendritic cells (FDC) of the tonsil of IgAN patients, but not in FDC of non-IgAN controls. On the other hand, IgA2, IgG, IgM and C3 did not show any differences in distribution between the two groups. Moreover, the expression of decay-accelerating factor (DAF), an inhibitor of homologous complement activation, and transforming growth factor-beta I (TGF-/β1). an inducer of antibody-producing ceils to IgA class switching, in FDC and interdigitating dendritic cells of the tonsil, respectively, which was also clarified in this study for the first time, was found to be identically distributed in the two groups. These findings may support the idea that IgA1. possibly in an immune complex form, is trapped by FDC and plays an important role in the persistent activation of particular B cell repertoires responsible for ihe onset and/or progression of IgAN.
T cell receptor gene rearrangements and cytotoxic activities of clones isolated from tumour-infiltrating lymphocytes (TIL) from melanoma patients
Authors: PANDOLFI F , BOYLE L A , TRENTIN L , OLIVA A , KURNICK J T 
Published: July 2010

SUMMARYThe lymphocytes which infiltrate tumours and are grown in vitro to be used in adoptive immunotherapy are often characterized by dominant rearrangement of their T cell receptor (TCR) genes. To investigate the frequency and function of cells contributing to the ‘dominant’ rearrangement, we have cloned two bulk cell lines of TIL derived from melanoma patients (TIL-1 and TlL-5). These IL-2-propagaled TIL cell lines had a CD8+ phenotype and exerted strong cytotoxic activity against autologous melanoma cells, but not against the natural killer (NK)-sensitive K-562 cell line or LAK targets such as Daudi cells. We derived 40 clones from TIL-1 and 23 from TIL-5. All tested clones were CD3+, CD4−, CD8+ and expressed the α/β TCR. From TIL-1.27 of 40 clones, and 13/19 of the TIL-5 clones lysed autologous tumour cells. In contrast to the NK, -negative bulk cultures, K-562 killing was detected in 21 of the TIL-1 clones and 17 of the TIL-5 clones. TIL-1 contained eight clones and TiL-5 two clones with lytic capacity against neither autologous tumour cells nor the K562 cell line, although these clones possessed lytic potential as evidenced in a lectimediated lysis assay. LAK activity was not detected in most clones. Cytotoxic activity against autologous tumour could be inhibited by preincubation with anti-CD3 or anti-HLA class I MoAbs, Of the 34 TlL-1 clones analysed, 15 shared a rearranged TCRβ EcoR1 restriction fragment of approximately 9 5 kb with the bulk culture. Clones sharing the EcoR1 10 5-kb dominant band present in TIL-5 bulk culture were also isolated. When the pattern of TCRβ rearrangement was compared with the cytotoxic functions, the following conclusions could be drawn: (i) clones contributing to the dominant band had heterogeneous functions. Most killed autologous tumour cells, but clones with no cytotoxic activity or even with no proliferative capacity in response lo autologous tumour cells were also detected among those contributing to the dominant rearrangement; (ii) some clones that share an apparently identical rearranged band different from the “dominant” rearrangement, may demonstrate the same cytotoxic function. In addition, our data suggest that many of the clones that share the dominant rearrangement originated from diverse progenitors. The high frequency of clonally diverse anti-tumour reactive TIL is likely to be a reflection of the in vivo selection of the TCR repertoire at the site of tumour. Further study of the TCR gene rearrangements should help to clarify how selection at this level can benefit future immunotherapeutic approaches.
Soluble intercellular adhesion molecule-1 (ICAM-1) in sera and bronchoalveolar lavage fluid of patients with idiopathic pulmonary fibrosis and pulmonary sarcoidosis
Authors: SHIJUBO N , IMAI K , SHIGEHARA K , HONDA Y , KOBA H , TSUJISAKI M , HINODA Y , YACHI A , OHMICHI M , HIRAGA Y , ABE S 
Published: July 2010

SUMMARYICAM-1 plays an important role in inflammatory diseases. To assess level of soluble ICAM-1 in the circulation and inflamed lesions, we measured levels of soluble ICAM-1 in the circulation and bronchial veolar lavage fluid (BALF) of patients with idiopathic pulmonary fibrosis (IPF) and with pulmonary sarcoidosis (PS) and of healthy volunteers (HV), and we also analysed ICAM-I expression of BALF cells in some patients and HV. IPF patients had significantly higher levels of circulating ICAM-1 than HV, while PS patients did not. By contrast, significantly increased levels of BALF soluble ICAM-1 were found in PS patients compared with those of HV, but not in IPF patients. There were no significant differences in the proportions of ICAM-1+ BALF lymphocytes in IPF patients, PS patients and HV. whereas significantly increased proportions of ICAM-1 pulmonary alveolar macrophages were found in PS patients compared with those of HV, but not in IPF patients. There was a significant positive correlation of BALF soluble ICAM-1 levels to BALF lymphocyte proportions in PS patients. Although the source of BALF soluble ICAM-1 is unclear. BALF soluble ICAM-1 appears to reflect the grade of local activity of sarcoidosis. An interesting discrepancy between soluble ICAM-1 levels in the circulation and BALF was found in IPF patients, and this might be an important clue to an understanding of this disorder.
Regulation of C3 and factor H synthesis of human glomerular mesangial cells by IL-1 and interferon-gamma
Authors: VAN DEN DOBBELSTEEN M E A , VERHASSELT V , KAASHOEK J G J , TIMMERMAN J J , SCHROEIJERS W E M , VERWEIJ C L , VAN DER WOUDE F J , VAN ES L A , DAHA M R 
Published: July 2010

SUMMARYPrevious reports have shown production of complement components C4. C2 and factor B by renal tissue. We have shown recently that human proximal tubular epithelial cells (PTEC) synthesize C3 in vitro, and that IL-2 enhances this production. In the present study we demonstrate that human mesangial cells (MC) in culture produce factor H and that supernatants of activated peripheral blood mononuclear cells (T cell growth factor (TCGF)) induce C3 production and enhance factor H synthesis in both a time- and dose-dependent manner. To investigate whether certain defined cytokines from TCGE were responsible for the observed effect., we tested various cytokines for their effect on complement production by MC. It is shown that IL-1 induces C3 synthesis whereas factor H production is up-regulated by TFN-γ, in both a dose- and time-dependent manner. Antibody blocking experiments revealed that C3 synthesis induced by both TCGF and IL-I could be blocked with antibodies specific for IL-I, and also that TCGF and IFN-γ enhanced factor H synthesis could both be blocked with antibodies specific for IFN-γ. Cycloheximide was able to inhibit C3 and factor H production, suggesting de novo synthesis of the proteins. mRNA-polymerase chain reaction (PCR) analysis revealed mRNA encoding for C3 after stimulation with TCG Fand IL-I. Factor H genes are constitutively expressed in cultured mesangial cells and its expression is up-regulated by TCGF and IFN-γ. Northern blot analysis with specific probes for C3 and factor H revealed bands which support the results obtained by PCR analysis.
Peripheral T lymphocyte depletion by apoptosis after CD4 ligationin vivo: selective loss of CD44− and ‘activating’ memory T cells
Authors: HOWIE S E M , SOMMERFIELD A J , GRAY E , HARRISON D J 
Published: July 2010

SUMMARYWe have demonstrated that a single intravenous bolus of rat anti-CD4 MoAb caused a small but prolonged increase in apoptosis in murine lymph nodes. We have quantified this process using the novel Highly Optimized Microscope Environment (HOME) interactive image analysis system and shown that the increase in apoptosis was sufficient to account for the observed depiction of the peripheral CD4+ T cell subset. This occurred in the absence of any other exogenous signal. Furthermore, there was no evidence of an inflammatory or necrotic response in the tissues, indicating that this was unlikely to be He or complement-mediated antibody killing. The anti-CD4-induced depiction selectively removed CD44− T cells. Using mice previously immunized with yeast-derived HIV-1 p24 recombinant protein there was sparing of memory T cell function after in vivo anti-CD4 treatment, except during a window of less than 24 h duration, when simultaneous exposure to antigen and anti-CD4 antibody resulted in the depiction of specific memory T lymphocyte function. This indicated that a very minor alteration in the frequency of apoptosis had a marked effect on cell number over time, and suggested that opportunistic infection associated with CD4+ T cell depletion may be explained by loss of memory cells when there is antigenic stimulation at the same time as CD4 ligation. These results have implications for the pathology of HIV-associated disease which is associated with ligation of CD4 molecules in vivo.
Anti-endothelial cell antibodies in sera of patients with autoimmune diseases: comparison between ELISA and FACS analysis
Authors: WESTPHAL J R , BOERBOOMS A M T H , SCHALKWUK C J M , KWAST H , DE WEIJERT M , JACOBS C , VIERWINDEN G , RUITER D J , VAN DE PUTTE I B A , DE WAAL R M W 
Published: July 2010

SUMMARYIn some patients suffering from rheumatoid arthritis (RA), vasculitis is a clear clinical manifestation, mentioned as rheumatoid vasculitis (RV). Autoantibodies directed against endothelial cells (AEA) have been implicated in the pathogenesis of this disorder, and it has been suggested in a number of studies that testing for AEA should be included in diagnosing RV. To test this hypothesis, we have evaluated the presence of AEA in sera of patients suffering from various autoimmune diseases, employing an ELISA with fixed cultured endothelial cells (EC). In ail the groups of patients ELISA-positive sera were present. A significant difference in percentage of positivity was found between the RA and RV group (P &lt;0.05). In addition, our results indicated that not only antibodies directed against antigens on the EC membrane were detected, but also antibodies directed against intracellular components like DNA. histones and cytoskeletal components. Therefore, we also tested all these patient sera on unfixed intact EC using indirect immunofluorescence followed by FACS analysis. Whereas in the total patient population 34 out of 65 patients were AEA-positive as determined in the ELISA, only seven patients were weakly positive when examined by flow cytometry. We conclude that: (i) an ELISA on fixed EC does not specifically detect AEA, A positive test result is. however, to some extent correlated with the occurrence of vasculitis, and may therefore be helpful in diagnosing this disease: (ii) EACS analysis is a more suitable method than ELISA to measure the presence of membrane-specific AEA in patient sera; (iii) specific IgG-AEA are less common in patients suffering from autoimmune disorders than was assumed previously.