bluuebunny/crossref_metadata_2025 huggingface dataset:
Interaction of papain-digested HLA class I molecules with human alloreactive cytotoxic T lymphocytes (CTL)
Authors: HAUSMANN R , ZAVAZAVA N , STEINMANN J , MÜLLER-RUCHHOLTZ W
Published: July 2010

Acute immunological rejection events of transplanted allogeneie organs are strongly dependent on T cell reactivity against foreign MHC products. The recognition requirements of alloreactive cytotoxic T cells arc of particular interest for finding approaches to modulating allorcactivity. The role of the allogeneic MHC molecule itself and/or an associated peptide in the interaction with the T cell receptor is still, however, unclear. Our studies have focused on the interactions of papain-digested HLA class I molecules with alloreactivc CD8+ CTL. These polypeptidcs, consisting of the polymorphic α1 and α2 and the monomorphic α3 domains, were used in both soluble and immobilized form to study their functional effects on anti-HLA-A2 reactive CTL. Purified polypeptides were of molecular mass 32–34 kD. HLA-A2 polypcptides (0–55 μg/ml) in soluble form induced half-maximal reduction of CTL cytotoxicity. These concentrations were quantitatively comparable to the effective doses of intact HLA class I molecules, which contain the hydrophobic transmembrane domain and the intracytoplasmic tail. In addition, specific activation requirements of these CTL were investigated in a serine esterasc release assay. Maximal degranulation was observed after 2 h of antigen contact. Purified HLA class I molecules allospccifically activated the anti-HLA-A2 CTL to degranulate serine esterase, when immobilized on plastic microtitre plates. Thus, polypeptidcs containing the polymorphic αl and α2 domains of human class I molecules potentially modulate the cytotoxic T cell response. This might have implications for the reduction or prevention of allograft rejection in recipients of foreign organs.
CR3 (CD11b, CD18): a phagocyte and NK cell membrane receptor with multiple ligand specificities and functions
Authors: ROSS G D , VĚTVIČKA V
Published: July 2010

The C3 receptor CR3 is expressed on phagocytie cells, minor subsets of B and T eells, and natural killer (NK) cells. It has important functions both as an adhesion molecule and a membrane receptor mediating recognition of diverse ligands such as intercellular adhesion molecule-1 (ICAM-1) and fixed iC3b. The receptor is capable of undergoing an activation event that regulates both its specificity for various ligands and its ability to mediate phagocytosis or extracellular eytotoxieily. Certain bacteria express earbohydrates or lipopolysaccharidcs (LPS) that can bind to and activate CR3, allowing the receptor to assume its activated state. Soluble β-glucan derived frotn the yeast Saccharomyces cerevisiae is a particularly potent stimulator of CR3, and produces an activated state of the receptor that permits neutrophil phagocytosis of iC3b-coated erytbrocytes or NK cell eytotoxicity of iC3b-coated tumour cells, tbat are normally resistant to NK cells.
Failure to detect hepatitis C virus (HCV) genome by polymerase chain reaction in human anti-HCV-positive intravenous immunoglobulins
Authors: DAMMACCO F , SANSONNO D , BEARDSLEY A , GOWANS E J
Published: July 2010

The prevalence of HCV antibodies was determined by a second-generation ELISA and a four-antigen rccombinant immunoblot assay in nine intravenous immunoglobulin (IVIG) preparations commercially available in Italy. In addition, the clinical safety of six of them was ascertained by polymerase chain reaction (PCR) of HCV RNA and a prospective study in 14 patients with immunodeficiency disorders. Results indicated that all IVIG preparations were anti-HCV-positive. However, there were substantial variations in their anti-HCV antibody titres. The preparations retained IgG subelass reactivilies to HCV-associated structural (C223) and non-structural (C33c, C100-3) proteins. Our sensitive and specific PCR assay was unable to detect HCV RNA in the six preparations tested. Clinical surveillance of IVIG-trcated patients prospectivcly evaluated over a mean period of 8·3 months failed to delect clinical and/or biochemical evidence of hepatitis.
Expression of CD5 and CD72 on T and B cell subsets in rheumatoid arthritis and Sjögren's syndrome
Authors: JAMIN C , LAMOUR A , PENNEC Y L , HIRN M , Le GOFF P , YOUINOU P
Published: July 2010

A minority of B cells express the CDS marker, which is found on virtually all T cells, and CD72 has been defined as the CD5 ligand on the B cell membrane. The mean fluorescence intensity (MFI) of the CD5 molecules was shown to be higher on CD4+ CD29+ than CD4+ CD45RA+ in peripheral blood (PB) and synovial fluid (SF) of rheumatoid arthritis (RA) patients (P < 0.0001 and < 0.001), and PB of Sjogren's syndrome (SS) patients and normal controls (P < 0·02 and < 0·03). This MFI declined once the CD4 expressed HLA-DR in PB of SS patients (P 0·004) and normal controls (P 0·02) or CD25 in PB of RA (P < 0·004) and SS patients (P < 0·0004). There was a correlation between the CDS MFI on CD4+ CD45RA+ and CD4+ CD29+ in RA (P 0·001) as well as SS (P 0·0007) PB. The CD72 MFI was impressively higher on CD5+ than CD5 B cells in PB and SF of RA patients (P < 0·0001 and P < 0·005) and PB of SS patients (P < 0·005) and normal controls (P < 0·005). Our data suggest that, in association with CD4CD29, CD5 is involved in CD5+B/CD5+ B cell interactions in non-organ-specific autoimmune diseases.
C1 inhibitor functional deficiency in systemic lupus erythematosus (SLE)
Authors: JAZWINSKA E C , GATENBY P A , DUNCKLEY H , SERJEANTSON S W
Published: July 2010

Cl inhibitor (Cl-inh) was assayed in eight SLE patients presenting with consistently low levels of intact C4. Cl-inh antigenic levels were normal in all patients; however, the function of the Cl-inh tested against C1 s and C1 r was variable and outside the normal functional range in seven of the eight patients. The molecular weight of patients’ Cl-inh protein was 105 kD, corresponding to the size of the intact molecule. The Cl-inh gene was analysed in all patients. Restriction fragments generated with TaqI, PstI and HgiAI gave no indication of a major Cl-inh gene rearrangement. Direct genomic sequencing of exon VIII revealed three polymorphic point mutations, but there were no changes from the normal gene in or around the reactive-centre residue of Cl-inh. Furthermore, we found no evidence for a C1-inh autoantibody in patients which could affect normal Cl-inh function in vitro. These results indicate that the etiology of Cl-inh dysfunction in SLE is heterogeneous and distinct from that reported in either hereditary or acquired angioedema.
Soluble intercellular adhesion molecule-1 (sICAM-1) in sera of patients with Graves’ ophthalmopathy and thyroid diseases
Authors: HEUFELDER A E , BAHN R S
Published: July 2010

Intercellular adhesion molecule, a ligand for the leucocyte integrins CDlla/CD18 (LFA-1) and CD11b/CD18 (Mac-1), that plays an important role in a variety of inflammatory and immune-mediated mechanisms, is strongly expressed in rctroocular connective tissue from patients with Graves' ophthalmopathy (GO) and involved in lymphocyte attachment to cultured retroocular fibroblasts via the ICAM-1/LFA-1-mediated pathway. Here, we report the detection and functional activity of a soluble form of the ICAM-1 molecule (sICAM-1) in sera from patients with GO and other thyroid diseases. Serum concentrations for sICAM-1 were determined using a highly sensitive ELISA. Compared with normal controls, patients with hyperthyroid or euthyroid GO and patients with Riedcl's invasive fibrous thyroiditis revealed markedly elevated sICAM-1 serum concentrations (all P 0·0001). In patients with Graves’ disease (GD) without clinical GO and in patients with Hashimoto's thyroiditis (HT), sICAM-1 levels were elevated to a lesser degree (both P < 0·001). sICAVI-l serum levels in patients with non-autoimmune hyperthyroidism due to a toxic adenoma were not significantly different from normal controls. In a separate group of 12 patients with severe inflammatory GO, sICAM-1 serum levels markedly declined (P < 0·0001) within 3 months of glucocorticoid therapy in nine patients who responded to this form of treatment with a decrease in periorbital inflammation. In contrast, sICAM-1 serum levels remained unchanged in three patients with poor response to steroids and persistent inflammatory periorbital disease. When tested in a cell adhesion assay, GO sera containing elevated concentrations of sICAM-1 were found to enhance the attachment of peripheral blood mononuclear cells (PBMC) to interferon-gamma (IFN-γ)-treated retroocular fibroblasts in a dose-dependent manner, up to a maximal stimulation of approximately 5×5-fold (P 0·001). This effect was abolished by preabsorption of sera with a MoAb against ICAM-1 and inhibited, in a dose-dependent manner, by coincubation with increasing concentrations of purified sICAM-1. In conclusion, sICAM-1 concentrations are markedly elevated in sera from patients with GO, and changes in sICAM-1 serum levels during glucocorticoid therapy closely parallel changes in the degree of inflammation. Given the capacity of sICAM-l to modulate the adhesion of lymphocytes to retroocular fibroblasts in vitro, sICAM-1 may play a role in the ongoing immune process within the connective tissue in GO.
Sera from patients with tuberculosis recognize the M2a-epitope (E2-subunit of pyruvate dehydrogenase) specific for primary biliary cirrhosis
Authors: KLEIN R , WIEBEL M , ENGELHART S , BERG P A
Published: July 2010

Anti-M2 antibodies in primary biliary cirrhosis (PBC) have been shown to react with the alpha-ketoacid dehydrogenase complex of the inner mitochondria! membrane consisting of six epitopes (E2 subunit of the pyruvate dehydrogenase complex (PDC). 70 kD; protein X of the PDC, 56 kD; alpha-ketoglutarate dehydrogenase complex, 52 kD; branched-chain alpha-kctoacid dehydrogenase, 52 kD; Elalpha subunit of PDC, 45 kD; and Elbeta-subunit of PDC, 36 kD). These epilopes are also present in the M2 fraction which is a chloroform extract from beef heart mitochondria. The E2 subunit of the PDC at 70 kD (M2a), especially, is a major target epitope which is recognized by about 85% of all PBC sera. However, analysing sera from 28 patients with active pulmonary tuberculosis it became evident that 12 (43%) also recognized the PDC-E2 subunit (M2a), as shown by Western blotting using the M2 fraction, the purified PDC, and the rccombinant PDC-E2. In contrast, only two of 82 patients with other bacterial and viral infections including 25 patients with Escherichia coli infections reacted with the PBC-specific epilope at 70 kD. Naturally occurring mitochondrial antibodies (NOM A) were present in 54% of the patients with tuberculosis and in 50% of patients with other infectious disorders. They recognized cither a determinant at 65 kD (epsilon) or at 60/55 kD (zeta/eta). None of the sera from 100 blood donors had anti-M2 but 14 had NOM A. Testing anti-M2 and NOMA-positive marker sera by Western blotting against membrane fractions derived from mycobacteria and E. coli it could be shown that-like mammalian mitochondria—they contain both the PBC-specific M2 antigen as well as the non-PBC-specific naturally occurring mitochondrial antigen system (NOM Ag). The observation that PBC-specific antibodies were preferentially induced in patients suffering from a mycobactcrial infection may provide some new clues to the still unknown etiology of PBC.
Increased limb involvement in murine collagen-induced arthritis following treatment with anti-interferon-gamma
Authors: WILLIAMS R O , WILLIAMS D G , FELDMAISN M , MAINI R N
Published: July 2010

We have tested the effect of administering H22, a hamster neutralizing MoAb to murine interferon-gamma (IFN-γ) in collagen-induced arthritis. Mice were initTtuni7ed with human type II collagen in adjuvant on day 1 and boosted with soluble collagen on day 21. H22 was administered (250 μg. intraperitoneally) either during the induction of arthritis (on days 0, 6, 13 and 20) or around the time of discase manifestation (on days 21, 28, 35 and 42). Control mice received either an isotypc-matched non-neutralizing MoAb or saline. Both treatment regimes gave sitnilar results. Treattncnt with H22 did not signifieantly affect the incidence of arthritis, time of onset, degree of oedetiia. histopathologi-cal severity, or level of anti-type II collagen IgG. However, a highly signiticanl increase (P < 0·01) in the number of limbs affected by arthritis was observed in the H22-treated group, irrespective of whether the antibody was administered during the induction of arthritis, or during the time of clinical manifcstalion of disease. From these results it was concluded that anti-IFN-γ treatment caused an inerease in the niitnber of arthritic lesions, but did not affect the severity of each individual lesion.
Inhibition by glucocorticoid and staurosporine of IL-4-dependent CD23 production in B lymphocytes is reversed on engaging CD40
Authors: KATIRA A , KNOX K A , FINNEY M , MICHELL R H , WAKELAM M , GORDON J
Published: July 2010

IL-4 synergizes with signals delivered through CD40 both for the induction of CD23/FcRH expression and for IgE synthesis. Moreover, engagement of CD40 on the B cell surface by MoAb overcomes the ability of intcrferons. transforming growth factor-beta, or anti-CD 19 to inhibit IL-4-dependent change. We now report that occupancy of CD40 relieves potent suppression of IL-4-induced CD23 production by glucocorticoid or the relatively broad-acting kinase inhibitor staurosporine. Interruption of the IL-4 signal was observed with concentrations of staurosporine considered to be selective for protein kinase C (PKC) inhibition (IC50= 10 nm) but not with genistein or tyrphostins. effective inhibitors of tyrosine kinase activity. On ligation of CD40, staurosporine no longer inhibited the IL-4 signal: at concentrations of between I and 20 nM. staurosporine actually increased by as much as 100% the rate of CD23 production stimulated on simultaneous activation through CD40 and 1L-4R. Such augmentation was not observed when the more specific PKC inhibitor RO-31-8220 was used; indeed. CD40 engagement was unable to overcome the ability of this inhibitor to block IL-4-promoted CD23 induction (IC50= 10 μm). Occupancy of CD40 did, however, thwart completely the usual ability of prednisolone to inhibit the IL-4 signal leading to CD23 induction. Activation through CD40 left inhibition of phorbol ester-induced CD23 expression by staurosporine, RO-31-8220, or glucocorticoid unchecked. These findings further highlight the intimate level of cross-talk existing between CD40 and IL-4R on resting B lymphocytes to promote CD23 expression, a phenotypic change which preludes IgE synthesis.
Antibodies to the CD5 molecule in normal human immunoglobulins for therapeutic use (intravenous immunoglobulins, IVIg)
Authors: VASSILEV T , GELIN C , KAVERI S V , ZILBER M-T , BOUMSELL L , KAZATCHKINE M D
Published: July 2010

TVIg are increasingly used Tor the treatment of autoimmune diseases. In the present study, we show thai IVIg contain antibodies directed againsl CD5, a cell surface molecule of T cells which is also a marker of the autoantibody-producing CD20+(‘B-r) subset of B lymphocyies, Antibodies to the CD5 molecule were demonstrated in IVIg by the ability of therapeutie preparations of IVIg to inhibit the binding of labelled CD5 MoAh to ihe CD5-expressing human Tcell lineH9. Prcincubation of H9 cells with IVIg or with F(ab')2 fragments prepared from IVIg resulted in dose-dependent inhibition nf the binding of CDS antibody. The presence in IVIg of antibodies to the CD5 molecule was fiirther confirmed by the binding of IVIg to mouse L cells that expressed human CD5 molecules following a stable transfeetion with CD5 cDNA. Human CD5 antibodies in IVIg provide therapeutic immunoglobulin preparations with the potential of modulating Tcell functions through CD5. and of regulating the expression of B cell subsets expressing CD5. This may have impliealions for the treatment of autoimmune diseases.
Anti-LKM-1 antibodies determined by use of recombinant P450 2D6 in ELISA and Western blot and their association with anti-HCV and HCV-RNA
Authors: SEELIG R , RENZ M , BÜNGER G , SCHRÖTER H , SEELIG H P
Published: July 2010

Several subtypes of anti-liver-kidney microsome antibodies (LKM) are known, LKM-1 antibodies associated with autoimmune chronic active hepatitis recognize P450 2D6, a cytochrome P450 monooxygenase. The frequent association of anti-LKM-l antibodies and hepatitis C virus (HCV) infections and the probable existence of an infectious and autoimmune form of anti-LKM-l-associated hepatitis, requiring different therapeutical strategies, necessitates the exact determination of anti-LKM-1 specificities. Therefore, we compared various antibody tests (immunofluorescence. ELISA with recombinant P450 2D6, and Western blot with recombinant and natural antigens and agargel double diffusion) with sera of 27 anti-LKM-1-positive chronic active hepatitis (CAH) patients, with 61 sera harbouring anti-mitoehondrial antibodies, 100 sera each from HCV-RNA-positive and HCV-RNA-negative patients, and 50 sera of healthy persons. Western blot techniques using recombinant MS2-polymcrase P450 2D6 fusion protein were found to be the most sensitive and specific method for anti-LKM-1 antibody determination in routine laboratory. The recently recognized association of anti-LKM-l antibody and HCV infection was confirmed by the results of this study. In anti-HCV and HCV-RNA-positive patients with anti-LKM-1 antibodies there was a preponderance of males with higher mean age and lower antibody titres. The results support the hypothesis of the existence of an autoimmune as well as an infectious (HCV triggered) subgroup of anti-LKM-l-positive hepatitis.
The B cell repertoire of patients with rheumatoid arthritis. Frequencies and specificities of peripheral blood B cells reacting with human IgG, human collagens, a mycobacterial heat shock protein and other antigens
Authors: RUDOLPHI U , HOHLBAUM A , LANG B , PETER H H , MELCHERS I
Published: July 2010

Using a potent in vitro limiting dilution culture system, we have activated human peripheral blood B cells to proliferate and to differentiate into antibody-secreting cells (ASC). Under these conditions 25–100 % of B cells are clonally expanded and produce IgM, IgG or IgA. Culture supernatants were tested for antibodies binding to human IgG-Fc fragments (RF), the 65-kD heat shock protein of Mycobacterium bavis (hsp60), human collagens type I, II, IV, V, transferrin, lactoferrin, albumins, and gelatine. All blood samples contained precursors of ASC (p-ASC) able to produce IgM binding to these antigens in frequencies above 003% of B cells. Most interestingly, a significant difference exists between rheumatoid arthritis (RA) patients and controls, concerning the relative frequencies of p-ASC able to produce monospecific or multircactive RF. Whereas most p-ASC(RF) in RA patients are monospecific (mean ratio 3.7), most p-ASC(RF) in healthy control persons are cross-reactive with at least one of five other antigens tested (mean ratio 0.2). The data suggest a disease-specific expansion of p-ASC committed to the production of monospecific rheumatoid factors.
Correlative studies of rheumatoid factors and anti-viral antibodies in patients with rheumatoid arthritis
Authors: FERRARO A S , NEWKIRK M M
Published: July 2010

An analysis of the relationship between the immune response to ubiquitous herpes family viruses, namely Epstein-Barr virus (EBV), cytomegalovirus (CMV), and varicella-zoster virus (VZV) and the presence of rheumatoid factors (RF), which are autoantibodics characteristic of patients with rheumatoid arthritis (RA), was conducted. Antibody profiles (RF, anti-viral antibodies) were monitored in the serum of the RA patients, and in normal individuals. No patient was found to have circulating RF in the absence of anti-viral antibodies. When the patients and normal controls were subdivided according to the presence of serum RF, it was found that when RF were present, the frequency of anti-CMV antibodies, but not anti-EBV or anti-VZV antibodies, was significantly higher (P= 0.02) when compared with RF-negative individuals. The litres of anti-CMV but not anti-VZV antibodies were found to increase in the RA patients with disease duration. To see if these viruses could stimulate RF production in vitro, peripheral blood mononuclear cells (PBMC) isolated from the patients and normal controls were stimulated with viral antigens. PBMC from normal controls, but not from RA patients, appeared to be responsive to viral antigen stimulation and produced RF. These data suggest that the immune response to CMV, to a greater extent than to EBV or VZV, correlates with the presence of RF.
Loss of mucosal CD4 lymphocytes is an early feature of HIV infection
Authors: LIM S G , CONDEZ A , LEE C A , JOHNSON M A , ELIA C , POULTER L W
Published: July 2010

T cell subsets in the gut mucosa are distinct populations and their imbalance in HIV has specific implications in infection. Alterations in T cell subsets in duodenal biopsies were investigated in 17 asymptomatic HIV patients, 24 AIDS patients and 10 controls with non-ulcer dyspepsia. Immunohistochemistry and immunofluoresccnce using MoAbs to CD3, CD4, CDS, CD68, CD45R A, CD45RO and gp 120 were performed on frozen sections. In the lamina propria. there was a significant depletion of CD4+ cells at all stages of HIV, but the density of CDS lamina propria cells was increased. Intraepithelial lymphocytes were decreased in AIDS patients. There was a significant correlation between cellular density and mucosal CD3+ lymphocytes, and between mucosal CD3+ and CDS+ lymphocytes. Although mucosal CD4, CD45RO+‘memory’ cells were decreased, CD8.CD45RO+‘memory’ cells were increased. Mucosal CD4+ lymphocyte depletion occurred early in HIV, and thus their role in mucosal protection against opportunistic infection should be revised. Mucosal CD8+ lymphocytes initially increased, but decreased when CD4 blood counts were depleted, perhaps contributing to loss of host protection against infection. Intraepithelial lymphocyte depletion may also contribute to opportunistic infection.
Characterization of anti-acetylcholine receptor (AChR) antibodies from mice differing in susceptibility for experimental autoimmune myasthenia gravis (EAMG)
Authors: GRAUS Y M F , van BREDA VRIESMAN P J C , de BAETS M H
Published: July 2010

In the murine model for EAMG we investigated the relation between disease susceptibility and fine specificity of anti-AChR antibodies obtained from high susceptible C57BI/6 and low susceptible BALB/c mice after immunization with Torpedo acetylcholinc receptor (tAChR). Anti-AChR MoAbs with fine specificity for the main immunogenic region (MIR), the α-bungarotoxin (α-BT)/ acetylcholine binding sites and other extra- and intracellular epitopes were isolated from both mouse strains. In total, nine out of 38 MoAbs obtained from C57BI/6 mice were directed against extracellular epitopes on mouse AChR in contrast to only one out of 27 MoAbs from BALB/c mice. A difference in antibody repertoire may underlie the difference in pathogenic response observed between these mouse strains. These results indicate that strain-specific differences in disease susceptibility in murine EAMG may be related to differences in the available repertoire of potential pathogenic antibodies.
Rheumatoid factor production by mononuclear cells derived from different sites of patients with rheumatoid arthritis
Authors: OTTEN H G , DAHA M R , DOLHAIN R J E M , de ROOY H H , BREEDVELD F C
Published: July 2010

To investigate the origin of circulating rheumatoid factor (RF) and the relation between RF production at different sites in patients with rheumatoid arthritis (RA), mononuclear cells derived from bone marrow, synovium and peripheral blood of patients with RA were examined for the presence of plasma cells and for their capacity to produce RF and other immunoglobulins in vitro. Analysis of culture supernatants for the presence of immunoglobulins demonstrated that cells derived from bone marrow, synovium and peripheral blood were all found to be capable of producing every immunoglobuiin and RF isotype investigated. No significant correlations were found between concentrations of immunoglobulin isotypes produced by cells derived from different sites of one individual. Significant correlations were found, however, between concentrations of RF isotypes produced by cells derived from the three sites. These results indicate that the production of RF in the different compartments is not an autonomously regulated process. Mononuclear cells derived from bone marrow were found to be able to produce RF in similar quantities to cells dissociated from synovial tissue. In combination with the fact that circulating immunoglobulins are produced mainly in the bone marrow, this observation suggests that bone marrow is also a major source of circulating RF.
Synthesis of the third component of complement (C3) by human gastric cancer-derived cell lines
Authors: KITANO E , KITAMURA H
Published: July 2010

This is a study of complement components secreted by gastric cancer-derived cell lines (MKN28. MKN45, MKN74 and KATO-III), each of which has a different histological origin. Haemolytic activity of complement component was detected only in the culture supernatant of KATO-III (C2 activity) and in that of MKN45 (C5 activity). However, the third component of complement, C3, was detected by an ELISA assay in the supernatants of all cell lines. In our studies focusing on C3 production by these cell lines, we have found that: (i) tumour necrosis factor (TNF) induced an increase in the amount of secreted C3 in a dose- and time-dependent fashion; (ii) TNF (10 U/ml) stimulated C3 secretion by these cell lines to levels of 25.4–62.9 ng C3/106cells per 24 hours; (iii) C3 haemolytic activity was detected in supernatants of TNF-stimulated cell lines. The mean specific activities of C3 by TNF (10 U/ml)-stimulated cell lines were 1.2–5.6 × 10- effective molecules/ng (e.m./ng), when that of C3 in normal human serum (NHS) was 1.7 × 106 e.m./ng; (iv) de novo synthesis of C3 by these cell lines was demonstrated by the effect of cycloheximide and by the incorporation of 35S-methionine into secreted C3; (v) immunoblot analysis of culture supernatants indicated that secreted C3 was mainly composed of C3α and C3β chains, but pro-C3 was also present. These results, which show the de novo synthesis and secretion of C3 by all the tested gastric cancer-derived cell lines in response to TNF, suggest the possibility that C3 may be secreted in the gastric wall as part of its normal physiology, or as a result of tumour pathology, and thereby participate in local immune or inflammatory responses.
Evaluation ofin vivoimmune complex formation and complement activation in patients receiving intravenous Streptokinase
Authors: FREYSDOTTIR J , ORMARSDOTTIR S , SIGFUSSON A
Published: July 2010

The usefulness of several different methods for delecting immune complex formation and complement activation in the circulation were applied to samples from patients receiving intravenous Streptokinase therapy for myocardial infarction. Streptokinase is a foreign antigen and can cause immune reactions. We collected samples from 13 patients, before Streptokinase administration (baseline), at the end of infusion (I h), 12 h later and on day 7. We measured IgG containing immune complexes (IgG-IC), free C3d and antibodies to Streptokinase by ELISA. and CR1. C3d and C4d on erythrocytes by flow cytometric assay. Antibodies to Streptokinase are common, as all but two of the patients had measurable antibody levels. During Streptokinase treatment there was a drop in antibody levels, most prominent in those patients who had high baseline levels. At the same time increased levels of free C3d and erythrocyte-bound C3d were observed. After 12 h free C3d was usually back to baseline level, but C3d on erythrocytes was still raised. These data indicate the formation of Streptokinase immune complexes in patients with high Streptokinase antibody levels, and show that these complexes are cleared rapidly from the circulation, leaving more persistent signs of complement activation. We conclude that free C3d is a good indicator of ongoing complement activation, whereas C3d on erythrocytes indicates that complement activation has recently taken place.
Systemic immunization against IgA in immunoglobulin deficiency
Authors: MOCHIZUKI S , SMITH C I E , HÅLLGREN R , HAMMARSTRÖM L
Published: July 2010

The presence of scrum IgM and IgG antibodies against IgA is common among individuals with IgA deficiency. The route of immunization is still unknown, but it is possible that immunization occurs through the gut. We analysed anti-IgA antibody production in gastrointestinal lavage, saliva and breast milk from patients with IgA deficiency. In no case was there any evidence of local production of anti-IgA antibodies. Immunization may thus be due to exposure to endogenous IgA and therefore represent a ‘true’ autoimmune phenomenon which may possibly be involved in the pathogenesis of the disease.
Multifunctional effects of anticomplementary agent K-76 on carrageenan-induced colitis in the rabbit
Authors: KITANO A , MATSUMOTO T , NAKAMURA S , OSHITANI N , OBATA A , OKAWA K , INOUE Y , KOBAYASHI K , NAKURA H
Published: July 2010

In this study the effect of K-76. a sesquiterpcne compound with anticomplementary activity isolated from a fungus culture, on carrageenan-induced colitis was studied from biochemical, histological and immunohistopathological aspects. K-76 suppressed epithelial cell loss, crypt abscess formation, inflammatory cell infiltration, mucosal atrophy, and ulceration. Immunohistochcmical examination of the colonic mucosa showed that the number of IgG- and IgM-positive plasma cells and the staining intensity for IgG and C3 were increased in carrageenan-induced colitis, but these changes were inhibited by K-76. Besides, serum mucoprotcin concentrations and CH50 levels were lower in the animals treated with carrageenan alone. K-76 exerted multifunctional activity, although its mechanisms of action remain obscure.
Expression of adhesion molecules on infiltrating T cells in thyroid glands from patients with Graves' disease
Authors: ISHIKAWA N , EGUCHI K , UEKI Y , NAKASHIMA M , SHIMADA H , ITO K , NAGATAKI S
Published: July 2010

The present study was performed to elucidate the role of adhesion molecules in the pathogenesrs of Graves'disease. Peripheral blood and intrathyroidal mononuclear cells were obtained from 14 patients with Graves'disease. The expression of adhesion molecules and HLA-DR antigen on CD4+ cells and CD4+ cell subpopulations was analysed by the two- or three-colour immunofluorescence method. The expression of adhesion molecules including LFA-1α, LFA-l/β, CD2, VLA-4α and VLA-5α on CD4+ cells in the thyroid gland was markedly higher than that in peripheral blood. In peripheral blood CD4+ cell subsets, the CD4+CD45RO+ cell population had an enhanced expression of the adhesion molecules compared with the CD4+ CD45RA+ cell population. However, there was no significant difference in the expression of adhesion molecules by CD4+ cell populations and subsets between Graves'disease and healthy subjects. The thyroid gland from Graves'disease contained a higher percentage of CD4+ CD45RO+ cells and a lower percentage of CD4+ CD45RA+ cells. In intrathyroidal CD4+ cell subsets, the CD4+CD45RO+ cell population had an increased expression of LFA-1 and CD2 compared with the CD4+CD45RA+ cell population, but there was no significant difference in VLA-4 and VLA-5 expression between the two cell subsets. Furthermore, the expression of LFA-1 and CD2 on the CD4+CD45RO+ cell population in the thyroid was significantly higher than that in matched peripheral blood. A similar finding was also observed for the CD4+CD45RA+ cell population. The thyroid gland had an increased percentage of CD4+HLA-DR+ cells compared with matched or healthy peripheral blood. However, there was no significant difference in the percentage of HLA-DR+ cells in the thyroid gland between CD4+CD45RO+ cell and CD4+CD45RA+ cell populations. These results suggest that increased expression of adhesion molecules on CD4+ cells may be responsible for the migration of these cells into thyroid glands and cellular interactions between these cells and thyroid epithelial cells.
Characterization of a human monoclonal autoantibody directed to cardiolipin/β2 glycoprotein I produced by chronic lymphocytic leukaemia B cells
Authors: MARIETTE X , LEVY Y , DUBREUIL M-L , INTRATOR L , DANON F , BROUET J-C
Published: July 2010

We determined the specificity and sequence of immunoglobulin molecules synthesized by monoclonal B cells from a patient with chronic lymphocytic leukaemia (CLL) who presented with a number of clinical and biological autoimmune symptoms. Helerohybrids obtained by fusion of CLL cells with the mouse X63-Ag 8.653 myeloma produced IgMλ MoAbs directed to the cardiolipin/β2 glycoprotein I (β2GPI) complex and ssDNA. They were devoid of polyreactivity. Nuclcotide sequence analysis of the variable domain of the μ chain indicated the utilization of the VH4 71.2 gene or one allotypic variant, DXp4 and JH3 segments. The λ light chain used the single gene from the Vλ8 subfamily, Jλ3 and Cλ3 genes. The VH gene displayed 11 nucleotide changes in comparison with its putative germline counterpart. However, these nucleotide changes correspond to variations observed in other published VH4 sequences, suggesting gene polymorphism ralher than somatic mutation. Dxp4 and Jh3 were also in germline configuration. The Vl gene exhibited a single replacement mutation in CDRl. These data suggest that the monoclonal CLLB cells in this patient retained VH and VL genes in germline configuration although they secreted a pathogenic anti-cardiolipin antibody associated with clinical symptoms, vasculitis and thrombosis, which may be provoked by antibodies to the phospholipid/β2GPI complex.
Prevention of cyclophosphamide-induced diabetes in the NOD/WEHI mouse with deoxyspergualin
Authors: NICOLETTI F , BORGHI M O , MERONI P L , BARCELLINI W , FAIN C , DI MARCO R , MENTA R , SCHORLEMMER H U , BRUNO G , MAGRO G , GRASSO S
Published: July 2010

Ten out of 20 (50%) 17-week-old female NOD/WEHI mice developed an acute form of autoimmune diabetes when injected with two large doses of cyclophosphamide (CY), given at 14-day intervals. If these mice were treated under a prophylactic regimen with 2–5 mg/kg body weight per day of the novel immunosuppressant deoxyspergualin (DSP) the onset of diabetes was completely prevented. Moreover, DSP-treated animals showed reduced signs of pancreatic insulitis, had lower percentages of splenic lymphoid cells (SLC) expressing IL-2 receptors and Ly-6C antigens on their surfaces, and these cells released lower amounts of interferon-gamma (IFN) when stimulated in vitro. These data, providing evidence for the capacity of DSP to protect NOD/WEHI mice from experimental autoimmune diabetes and to modulate histo-immunological pathogenic pathways, indicate DSP as a drug of potential interest in the treatment of human insulin-dependent diabetes mellitus.
Immunization of mice with baculovirus-derived recombinant SV40 large tumour antigen induces protective tumour immunity to a lethal challenge with SV40-transformed cells
Authors: SHEARER M H , BRIGHT R K , LANFORD R E , KENNEDY R C
Published: July 2010

In this study, we examine the humoral immune responses and in vivo tumour immunity induced by baeulovirus reeombinant simian virus 40 (SV40) large tumour antigen (rSV40 T-ag). BALB/c mice immunized with rSV40 T-ag produced antibody responses that recognized SV40 large tumour antigen (T-ag) by ELISA. Analysis of these anti-SV40 T-ag responses indicated that the antibodies recognized epitopes associated with both the carboxy and amino terminus of SV40 T-ag. This pattern of SV40 T-ag epitope recognition was similar to that observed in anti-SV40 T-ag responses induced by inoculation with irradiated SV40-transformed cells. Mice immunized with either rSV40 T-ag or with the inactivated transformed cells were protected from a subsequent in vivo lethal tumour challenge with live SV40-transformed cells. These studies suggest that humoral immune responses induced by rSV40 T-ag are similar in epitope specificity to that induced by inactivated SV40-transformed cells. In addition, recombinant tumour-specific antigens from papovaviruses, such as SV40, can be used to induce tumour immunity which protects from a subsequent lethal tumour challenge. This study may provide insight into the use of recombinant tumour antigens as putative tumour vaccines and in the development of active immunotherapeutic strategies for treating virus-induced eancers.
Phenotypic analysis of cells in bronchoalveolar lavage fluid and peripheral blood of maedi visna-infected sheep
Authors: LUJÁN L , BEGARA I , COLLIE D D S , WATT N J
Published: July 2010

A phenolypic analysis of bronchoalveolar lavage fluid (BALF) and peripheral blood (PB) cells in maedi visna virus (MVVVinlecled sheep has been performed. The differential cell eount in BALF from MVV-infeeted animals was characterized by a significant increase (P < 0±05) i n lymphocytes and neutrophils. Lymphocyte phenotyping in BALF from MVV-infeeted sheep showed a significant decrease (P < 0·05) or CD4+ cells, a significant increase (P < 0·05) of CD8+ eells and a significant inversion (P<0·001) of the CD4+/CD8+ ratio. CD5+ lymphocytes were also significantly decreased (P < 0·05). γδ T cells and B cells did not differ significantly when compared with the controls. No correlation was observed between BALF and PB lymphocyte phenotypes. BALF macrophages from MVV-infeeted animals showed inereased MHC class II expression and BALF lymphocytes from the same animals demonstrated up-regulation of LFA-1 and LFA-3 expression. These findings and their relationship with lentiviral pathogenesis are discussed.
Differential mechanisms of intracellular killing ofMycobacterium aviumandListeria monocytogenesby activated human and murine macrophages. The role of nitric oxide
Authors: BERMUDEZ L E
Published: July 2010

Murine peritoneal macrophages activated with interferon-gamma (IFN-γ) produce large quantities of nitric oxide and are efficient in the killing of certain intracellular pathogens. To examine the role of this mechanism in the killing of Mycobacterium avium by murine and human macrophages, we infected mouse peritoneal macrophages and human monocyte-derived macrophages with M. avium and Listeria monocytogenes and stimulated the cells with recombinant tumour necrosis factor (TNF), granulocyte-macrophage colony-stimulating factor (GM-CSF) or IFN-γ, in the presence or absence of N-monomethyl-L-arginine (NMA) or arginase. Neither competitive inhibition with NMA nor depletion of arginine by arginase had any effect on the inhibition of growth/intracellular killing of M. avium by activated human and murine macrophages. In contrast, activation of murine but not human macrophages infected with L. monocytogenes by IFN-γ was significantly inhibited by the addition of NMA/arginase. Furthermore, murine macrophages produced large concentrations of nitric oxide following stimulation with recombinant cytokines, although no significant increase of nitric oxide production was observed with human monocyte-derived macrophages.
Direct evidence that human follicular dendritic cells (FDC) rescue germinal centre B cells from death by apoptosis
Authors: LINDHOUT E , MEVISSEN M L C M , KWEKKEBOOM J , TAGER J M , de GROOT C
Published: July 2010

It is supposed that FDC have a pivotal role in the rescue of gcrminal centre (GC) B lymphocytes from apoptosis. However, formal proof for this hypothesis has not as yet been presented. In the present study FDC and GCB cells were isolated from humati tonsils and cultured. When brought into culture FDC and B cells rapidly formed spherical clusters. T cells were not observed inside these clusters. At dilTerent time points cultures of FDC and B cells were siipravitally stained with Hoechst 33258 or aeridine orange and examined by direct observation tising fluorescence microscopy. Viable B cells appeared to be profoundly restricted to clusters, whereas cells not taking part in clusters all had an apoptotic appearance. The formation of clusters could be prevented by addition of MoAbs against CD11a (LFA-1α) or CD49d (VLA-4α). resulting in an apoptotic appearance of virtually al B lymphocytes. The present data demonstrate that a physical interaetion between FDC and germinal centre B lymphocytes is able to rescue the latter from apoptotic cell death.
Percentile ranges for serum IgG subclass concentrations in healthy Chinese children
Authors: LAU Y L , JONES B M , NG K W , YEUNG C Y
Published: July 2010

In order to establish normal reference ranges of serum immunoglobulin G subclasses for different age groups in Chinese, we measured the four IgG subclasses by radial immunodiffusion using poiyclonal antisera in 350 normal healthy subjects (148 males and 202 females) recruited Irom ihe community. There was no signifieant sex difference for all the four IgG subclasses. Using Box-Cox transformation, we constructed smooth age-dependent percentile curves lor the four IgG subclasses. For the 50th percentile values, the plateaus Tor IgGl lo lgG4 are respectively 970 mg/dl at 13 years old. 481 mg/dl al 18. 48 mg/dl at 17 and 80 mg/dl at 13. Methodology, reagents, environmental and genetic factors might be responsible for the observed difference among the various reports. The IgG2 level in our Chinese population seemed to be higher than that in Caucasians, which might account for the very low incidence of invasive Haemophilus influenzae type b disease among the Chinese. Our IgG l level also plateaued later and at a higher level than that in other studies.
Molecular analysis of C3 allotypes in patients with nephritic factor
Authors: FINN J E , MATHIESON P W
Published: July 2010

The autoantibody nephritic factor (NeF) leads to complement consumption in vivo and is associated with type II mesangiocapillary glomeruloncphritis(MCGN II) and partial lipodystrophy (PLD). The third component of complement (C3) exists in two common allotypic forms. C3S and C3F, distinguished at the protein level by electrophoresis. An increased frequency of the rarer C3F allele has been reported in several autoimmune conditions, including one small series of patients with NeF. However, patients with NeF have low levels of circulating C3 so that allotyping at the protein level is difficult. The molecular basis of the S/F polymorphism has recently been established: a single base change at the DNA level encodes a single amino acid substitution at the protein level. A second polymorphism, closely linked to the first, is defined by the MoAb HAV4–1, and is also due to a single base change. These polymorphisms can therefore be analysed at the DNA level. We have used the amplification refractory mutation system (ARMS), a modification of the polymerase chain reaction (PCR), to analyse these two C3 polymorphisms at the DNA level in 26 patients with NeF. The allele frequencies of C3S and C3F were 0·673 and 0·327 (predicted values 0·79 and 0·2, x2= 4·813, P < 0·05), giving a relative risk of 2·1 for the development of NeF conferred by the presence of a C3F allele. The HAV 4–1 allele frequencies were (-) 0·71 and (+)0·29. i.e. not significantly different than predicted from the linked C3S/F allele frequencies. This is the largest series of patients with NeF yet published, and our data confirm an association between C3F and NeF. Possible mechanisms for this link are discussed.
Serum markers of T cell activation in relapses of Wegener's granulomatosis
Authors: STEGEMAN C A , TERVAERT J W COHEN , HUITEMA M G , KALLENBERG C G M
Published: July 2010

Levels of soluble IL-2 receptor (sIL-2R). soluble CD4 (sCD4) and CDS (sCD8) were measured by sandwich ELISA as markers for T cell activation in serial serum samples from 16 patients showing 18 histologically proven relapses of Wegener's granulomatosis (WG). Levels of sIL-2R increased from 1065 U/ml (median, range 373–2345 U/ml) 6 months before the relapse to 1684 U/ml (median, range 486–3404 U/ml) at the moment of relapse for the whole group (P= 0·10). The eight major relapses showed a profound rise in sIL-2R levels, from 1008 U/ml (median, range 686–1553 U/ml) 6 months before the relapse, to 1994 U/ml (median, range 1469–3404 U/ml) at the moment of relapse (P < 0·01). The levels of sIL-2R at the moment of relapse were significantly higher at the eight major relapses than at the time of the 10 minor relapses (P<0·05). Minor relapses were not accompanied by a significant rise in sIL-2R levels. Titres of antineutrophil cytoplasmic antibodies (ANCA) rose by two or more titresteps or from negative to positive in 15/18 patients during the 6 months period before the relapse. In all seven cases with both a rise of the ANCA titre and an at least 25% increase in sIL-2R levels, the rise in ANCA preceded the rise in sIL-2R by at least I month. The level of slL-2R at the moment of relapse correlated with the level of C-rcactive protein (P= 0·488, P<0·05) and with the disease activity score (r= 0·824, P<0·002). There were no significant changes in levels of sCD4 or sCD8. although the levels of sCD4 tended to be higher at the time of major relapses. We conclude that major relapses of Wegener's granulomatosis arc accompanied by systemic T cell activation. T cell activation, however, does not appear to precede the rise in ANCA titre.
Gender-related influences on the development of chronic graft-versus-host disease-induced experimental lupus nephritis
Authors: TREURNIET R A , BERGIJK E C , BAELDE J J , DE HEER E , HOEDEMAEKER J , BRUIJN J A
Published: July 2010

Autoimmune diseases are far more common in women than in men. In the incidence of systemic lupus erythematosus (SLE), the female-to-male ratio is as high as 10:1. This suggests that sex hormones may play a fundamental role in determining the susceptibility to these diseases. In order to investigate the sex-related differences in the inducibility of chronic graft-versus-host disease-related experimental lupus nephritis, lymphocytes from female DBA/2 donor mice were administered to either male or female (C57BL10 × DBA/2)F1 recipients. An additional group of male recipients received lymphocytes from male DBA/2 donors. After four cell transfers, female recipients developed a significantly higher albuminuria than both male groups. Serum concentrations of autoantibodies against glomerular basement membrane (GBM). collagen IV, and laminin were significantly higher in females 2–4 weeks after induction. Levels of circulating autoantibodies against renal tubular epithelial antigens (RTE) and nuclear antigens were not different between the sexes. In transfer studies, the necessity of the presence of anti-GBM and anti-RTE autoantibodies for the development of glomerulonephritis was confirmed. These findings indicate that: (i) in this model of lupus nephritis, susceptibility to glomerulonephritis is strongly influenced by sex-related genes; and (ii) among the variety of autoantibodies occurring in this model of SLE, both anti-GBM and anti-RTE autoantibodies play a key role in the pathogenesis of glomerulonephritis.