bluuebunny/crossref_metadata_2025 huggingface dataset:
T and B cell responses following immunization with tetanus toxoid in IgA nephropathy
Authors: FORTUNE F , COURTEAU M , WILLIAMS D G , LEHNER T
Published: July 2010

The B and T cell responses were investigated in IgA nephropathy before and after immunization with tetanus toxoid (TT). Both IgA and IgG anti-tetanus toxoid antibodies were elicited, but the IgA ntibodies were significantly greater in patients (92·6 ± 11·7 ELISA units) than in the controls (49·2 ± 7·5 ELISA units). This was associated with a significantly greater proportion of IgA+ B cells in patients than controls before immunization. However, a significant increase in the proportion of IgA 1 binding CD4 and CD8 cells was also found. The proportion of CD3 cells with γδ T cell receptors (CD3+TCRγδ+), was significantly greater before immunization in the IgA nephropathy patients 37·0%± 2·4), compared with controls (100%± 2·3; P < 0·001). Immunization with TT further enhanced the CD3+TCRγδ5+ cells in patients to 45·8%± 7·2 compared with controls (16·3%± 4·5), with a corresponding decrease in CD3+TCRαβ+ cells in the patients (P < 0·001). CD3+TCRγδ+ cells are upregulated by common microbial antigens and clinical exacerbations of IgA nephropathy are frequently associated with mucosal infections and a rise in serum IgA concentration. The increased TCRγδ (5 expression may be responsible for the enhanced IgA antibody response in IgA nephropathy. The increase in IgA antibodies may then exert a controlling effect by binding to augmenting T cells and thereby inhibiting their function.
Autostimulatory effects of IL-6 on excessive B cell differentiation in patients with systemic lupus erythematosus: analysis of IL-6 production and IL-6R expression
Authors: KITANI A , HARA M , HIROSE T , HARIGAI M , SUZUKI K , KAWAKAMI M , AWAGUCHI Y K , HIDAK T , KAWAGOE A M , NAKAMURA H
Published: July 2010

Introducing avidin-biotin complex ELISA for anti-DNA antibody, the mechanism of in vitro production of anti-ssDNA antibody as well as of polyclonal immunoglobulin mediated by an IL-6-IL-6R loop was studied in patients with systemic lupus crythematosus (SLE). Regardless of the presence or absence of T cells, B cells from SLE patients could produce IgG anti-ssDN A antibody as well as total IgG without any stimulation. Low density B cells obtained by Percoll gradient density cenlrifugation responded to rIL-6 to produce IgG and IgG anti-ssDNA antibody. rIL-2 and rIL-4 had lesser effects on the differentiation of low density B cells. In fact, IL-6R was preferentially expressed on low density B cells from active SLE patients, as detected by anti-IL-6R MoAb, MT18, which did not inhibit IL-6 binding. SLE B cells, especially high density B cells, produced greater amounts of IL-6 in culture supernatants than did T cells, regardless of whether disease was active or inactive. Normal T cells and B cells did not produce significant amounts of IL-6. Thus, endogenous IL-6 produced by high density B cells bound to the IL-6R preferentially expressed on the low density B cells, and drove them into terminal differentiation, especially in active SLE patients. Further, addition of polyclonal anti-IL-6 or anti-IL-6R MoAb (PM1). which inhibited IL-6 binding, both inhibited IgG anti-ssDNA antibody as well as total IgG production by SLE B cells in a dose-dependent manner. These results suggest that interruption of the autocrine IL-6 loop would be of therapeutic value in SLE.
Bone marrow and peripheral blood natural killer cell activity in lymphomas. Its response to IL-2
Authors: CALDERA L H , LEON-PONTE M , ACQUATELLA G , BIANCO N E , BLANCA I
Published: July 2010

Natural killer (NK) cytotoxic activity was simultaneously investigated in bone marrow mononuclear cells (BMMC) and peripheral blood lymphocytes (PBL) from nine Hodgkin's disease (HD) and 15 non-Hodgkin lymphoma (NHL) untreated patients. Twenty-five PBL samples and seven bone marrow specimens from healthy individuals were also included as control group (C). NK cell activity was evaluated in basal condition and post-stimulation with human recombinanl IL-2 (rIL-2). Data were expressed in K values (number of BMMC or PBL needed to lyse 50% of the target cells). In basal condition, both HD and NHL patients showed a NK cell activity comparable to the C group, both in BMMC (HD, K=2·48±1·3; NHL, K=3·8±2·0; C, K=3·2±0·7) and PBL (HD, K=2·0±1·0; NHL, K=2·3±1·0; C, K= 2·2±0·2). Stimulation with rIL-2 induced a significant and comparable enhancement of the NK activity in PBL from HD, NHL and C while the response to rIL-2 of the BMMC in most of the HD and NHL patients was significantly greater than the C group. Responder cells were characterized by negative selection with specific MoAb plus complement as a CD3−, CD16+, CD56+ cytotoxic cell and further confirmed by flow cytometry. We postulate that IL-2 activation of bone marrow NK cell precursors, in addition to enhancing the activity of circulating NK, may be of value for the therapeutic rationale of IL-2 in patients with lymphoma.
Heterogeneous effects of exogenous IL-2 on HIV-specific cell-mediated immunity (CMI)
Authors: BELL S J D , COOPER D A , KEMP B E , DOHERTY R R , PENNY R
Published: July 2010

A characteristic feature associated with HIV-1 infection of the human host is a chronic decline in circulating CD4+ T helper/inducer cell numbers. Impaired cell-mediated immune functions usually occur in parallel with the decline in CD4+ T cells. Activated CD4+ T helper cells are a major source of endogenous IL-2 which is required for the immunoregulation of both antigen-specific B cells and CD8+ T cells. HIV-specific T cell proliferative responses are said to be weak and inconsistent, even during the asymptomatic phase of disease. We thus wished to determine how exogenous IL-2 affected HIV-specific T cell proliferation at different stages of the disease. Our cohort of 81 included both asymptomatic and symptomatic HIV-infected patients as well as uninfected normal donors. Proliferative responses of peripheral blood mononuclear cells (PBMC) that were elicited during culture with an immunodominant gp41-derived synthetic peptide, gp41{8}, and which were known to be CD8+ cell-associated in asymptomatics only, were used to analyse the effects of exogenous IL-2. IL-2 had three main effects on HIV-specific proliferation, namely (i) an additive effect, (ii) a synergistic effect, and (iii) an induced effect. More specifically, low dose exogenous IL-2 frequently augmented lymphoproliferation in both asymptomatic and symptomatic gp41{8} rcspondcrs. In most symptomatics, however, who were predominantly gp41{8} non-responders, exogenous IL-2 induced lymphoproliferation. Flow cytometric analyses using dual immunofluorescence were used to analyse the T cell subset distribution of proliferating PBMC cultures. During culture with gp41{8}, bothCD4+and CD8+ T cell numbers increased. However, after the addition of exogenous IL-2 to gp41{8}-containing cultures, CD8+ cell-associated lymphoproliferative responses were preferentially augmented. These results suggest that in symptomatics there is an inadequate supply of endogenous IL-2 to help maintain the strong and effective CD8+ cell-associated anti-viral immunity, and an exogenous supply of IL-2 may be required.
Activation of complement in normal serum by hydrogen peroxide and hydrogen peroxide-related oxygen radicals produced by activated neutrophils
Authors: SHINGU M , NONAKA S , NISHIMUKAI H , NOBUNAGA M , KITAMURA H , TOMO-OKA K
Published: July 2010

Neutrophils activated by soluble partculate stimuli generate superoxide anion and subsequently form hydrogen peroxide and other oxygen radicals. The effect of hydrogen peroxide on the complement system in normal serum was investigated. Treatment of normal serum with hydrogen peroxide resulted in a diminution of the haemolytic activity of the total and alternative complement pathways and the haemolytic titres of C3 and C5 but not of C2, in normal serum. These decreases in complement activity depended on the concentration of hydrogen peroxide added to the serum. Immunoelectrophoretic analysis of hydrogen peroxide-treated serum showed that C3 and C5 proteins were activated. Complement degradation products C3a and C5a were produced in normal serum treated with hydrogen peroxide, and 20 mm EDTA abolished C3a and C5a production in hydrogen peroxide-treated serum but 20 mm Mg-EGTA did not. Catalase completely abolished and dimethylsulphoxide and D-mannitol, hydroxyl radical scavengers, partially inhibited the hydrogen peroxide-mediated complement activation. Hypochlorite, incubated with normal serum, significantly inhibited serum haemolytic activity, and sodium thiosulphate, a reducing agent, abolished the effect of hypochlorite. Normal serum incubated with activated neutrophils showed neutrophil chemotactic activity and decreased serum haemolytic activity, and the addition of catalase or methionine (S mm) completely abolished the effects of activated neutrophils. These results suggest that hydrogen peroxide activates complement via an alternative pathway of complement activation and that hydroxyl radicals and other hydrogen peroxide-related species such as hypochlorite are most likely involved in hydrogen peroxide-mediated complement activation. Complement activation by oxygen radicals produced by activated neutrophils may be one of the mechanisms by which complement is activated in human immune complex diseases.
Detection of cytokines at the site of tuberculin-induced delayed-type hypersensitivity in man
Authors: CHU C Q , FIELD M , ANDREW E , HASKARD D , FELDMANN M , MAINI R N
Published: July 2010

Cytokines are chiefly local mediators which play an important role in the regulation of the cell-cell interactions which may be involved in the development of the delayed-type hypersensitivity (DTH) reaction. Using immunohistochemical techniques, the presence of IL-la, IL-lβ, IL-6, interferon-gamma (IFN-γ) and tumour necrosis factor-alpha (TNF-α) in the skin in tuberculin-purified protein derivative (PPD)-induced DTH reactions was investigated in six normal individuals. Cells staining for these cytokines were first observed 6 h after PPD challenge, and they were detected throughout the duration of the 7-day experiment. The number of cells staining for IFN-γ reached a peak at 48 h. where 33% of the total aggregate cells were positive, but declined thereafter to 3% at day 7. On the other hand, the number of cells staining for TNF-α and IL-1 persisted at high levels throughout the observation period of 7 days (e.g. at 48 h and thereafter, about 40% cells positive for TNF-α and 20% for IL-la and IL-1β). Double immunofluorescence and staining on sequential sections showed that IFN-γ-staining cells were CD3+ T cells; TNF-α, IL-1 and IL-6 staining cells were mainly of the CD68* macrophages/monocytes and that 80% of the CDla4 cells (Langerhans-like cells) in the dermis contained TNF-α and IL-1. The presence of these cytokines at the site of inflammation suggests that they may be locally produced by the inflammatory cells. Their persistence during the reaction suggests that they are intimately associated with this response, and are involved in the development of the reaction.
Lymphocytes from patients with rheumatoid arthritis produce agalactosylated IgG in vitro
Authors: BODMAN K B , SUMAR N , MACKENZIE L E , ISENBERG D A , HAY F C , ROITT I M , LYDYARD P M
Published: July 2010

The percentage of oligosaccharide chains lacking galactose was measured in IgG obtained from pokeweed mitogen-activated cultures of blood lymphocytes from patients with rheumatoid arthritis and controls. Secreted IgG from rheumatoid arthritis lymphocytes was deficient in galactose compared with IgG from the lymphocytes of controls. This confirms that agalactosylation is a significant feature of the disease and demonstrates that it can occur at the B cell level and is not merely a post-secretory event.
Interferon-alpha and dexamethasone inhibit adhesion of T cells to endothelial cells and synovial cells
Authors: EGUCHI K , KAWAKAMI A , NAKASHIMA M , IDA H , SAKITO S , MATSUOKA N , TERADA K , SAKAI M , KAWABE Y , FUKUDA T , ISHIMARU T , KUROUJI K , FUJITA N , AOYAGI T , MAEDA K , NAGATAKI S
Published: July 2010

We investigated whether interferon-gamma (IFN-γ), interferon-alpha (IFN-α) and glucocorticoids affected the adhesion of T cells to human umbilical endothelial cells or human synovial cells. About 30% of peripheral blood T cells could bind to unstimulated endothelial cells, but only a few T cells could bind to unstimulaled synovial cells. When both endothelial cells and synovial cells were cultured with recombinant IFN-γ (rlFN-γ), the percentage of T cell binding to both types of cells increased in a dose-dependent manner. rIFN-α and dexamethasone blocked the T cell binding to unstimulated endothelial cells. Furthermore, rIFN-α and dexamethasone suppressed T cell binding to both endothelial cells and synovial cells stimulated by IKN-γ, and also inhibited intercellular adhesion molecule- l (ICAM-1) expression on both endothelial cells and synovial cells stimulated by 1 FN-γ. These results suggest that IFN-α and glucocorticoids may inhibit T cell binding to endothelial cells or synovial cells by modulating adhesion molecule expression on these cells.
In vivoclearance by the mononuclear phagocyte system in humans: an overview of methods and their interpretation
Authors: HALMA C , DAHA M R , VAN ES L A
Published: July 2010

The mononuclear phagocyte system (MPS) is responsible for the elimination of foreign material, effete autologous material and immune complexes. To study the relationship between MPS function and human disease, several test substances have been developed, and used to determine the clearance capacity of the MPS in human subjects in vivo. These test substances and the multitude of factors that influence the elimination of these substances (and complicate the interpretation of the test results) are discussed. Use of these probes bas provided important new insights, that may lead to the development of treatment modalities by which MPS function is modified in order to influence disease processes more effectively.
Adhesion molecule expression in Graves' thyroid glands; potential relevance of granule membrane protein (GMP-140) and intercellular adhesion molecule-1 (ICAM-1) in the homing and antigen presentation processes
Authors: MIYAZAKI A , MIRAKIAN R , BOTTAZZO G F
Published: July 2010

To assess the potential role of adhesion molecules in the pathogenesis of Graves' disease, we examined the expression of several of these adhesion molecules, including intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule (VCAM-1) and granule membrane protein-140 (GMP-140), in sections of Graves' thyroid glands and control thyroids, using Immunohistochemical techniques. Up-regulated expression of GMP-140 was frequently observed on endothelial cells (EC) of post-capilliary venules in all Graves' thyroids examined, compared with an occasional weak staining on EC control glands. Some capillary EC around thyroid follicles (perifollicular EC) were strongly positive for GMP-140 in the Graves' thyroids in contrast to a negative staining on the same Structures in the control glands. In addition, there was a correlation between the reactivity and frequency of GMP-140 expression on EC and the severity of mononuclear cell (MNC) infiltration in the Graves' thyroids. The expression of ICAM-1 was up-regulated on perifollicular EC and EC of small venules in some thyroids of both Graves' and control groups. Conversely, no significant expression was observed on any type of EC for both endothelial-leucocyte adhesion molecule-1 (ELAM-1) and VCAM-1. However, dendritic-like cells, present within lymphocytic infiltrates, were positive for VCAM-1 in most of the Graves' thyroids examined, especially in those with a severe lymphocytic infiltration. Thyrocytes were constantly negative for the expression of all tour adhesion molecules investigated. These data suggest that GMP-140, as well as ICAM-1, could play an important role in the initiation of MNC infiltration in Graves' disease. ELAM-1 and VCAM-1 appear not to be relevant for the migration of MNC from the blood vessels into the target gland, although VCAM-1 expression on dendritic-like cells might play an additively tissue-selective role in autoantigen presentation and subsequent elicitation of autoimmune phenomena.
Distribution of retroviral p15E-related proteins in neoplastic and non-neoplastic human tissues, and their role in the regulation of the immune response
Authors: SCHEEREN R A , OOSTENDORP R A J , VAN DER BAAN S , KEEHNEN R M J , SCHEPER R J , MEIJER C J L M
Published: July 2010

In patients with head and neck carcinomas and in patients with chronic purulent upper airway infections, low molecular weight retroviral p15E-like factors are found. These factors are responsible for partial defects in the cellular immune response. We studied the distribution of these p15E-related proteins in neoplastic. inflamed and normal human tissues and related these findings with the presence of p15E-like factors in patients's era. Demonstration of p15E-like proteins in sera of patients with upper airway infections and of patients with head and neck carcinomas correlated exclusively with the presence of p15E in normal and pathologic epithelium of the upper respiratory tract, p15E was not demonstrated in epithelia of other localizations. Our results suggest that chronic stimulation or neoplastic transformation of the epithelia of the upper respiratory tract stimulates the production of p15E-like proteins leading to their reported immunosuppressive actions.
Secretion of Toxoplasmagondii-specfic antibodyin vitroby peripheral blood mononuclear cells as a new marker of acute toxoplasmosis
Authors: VENDRELL J P , PRATLONG F , DECOSTER A , BOULOT P , CONGE A M , DARCY F , SEGONDY M , HUGUET M F , SERRE A
Published: July 2010

Antigen-specific antibody secretion in vitro by peripheral blood mononuclear cells (PBMC) reflects an in vivo stimulation of the immune system by the antigen. Primary infection of immunocompetent patients with T. gondii causes an acute infection followed by chronic toxoplasmosis. We examined in vitro anti-Toxoplasma antibody production by PBMC during the acute and chronic phases of toxoplasmosis. PBMC from patients with acute or chronic toxoplasmosis and seronegative subjects were cultured for up to 6 days. Anti-Toxoplasma antibodies were assayed in supernatants by ELISA and immunoblotting. Anti-Toxoplasma antibodies were detected in supernatants of PBMC from 29 pregnant women who seroconverted during gestation. PBMC from 17 patients who had chronic toxoplasmosis and PBMC from 10 seronegative healthy controls did not secrete Toxoplasma-specific antibodies. This in vitro antibody secretion was spontaneous, active and transient since it disappeared between 11 and 24 weeks after seroconversion. Anti-Toxoplasma antibody secretion by PBMC from patients with acute toxoplasmosis is consistent with an in vivo stimulation of the immune system by T. gondii antigens. Our results represent a new approach for studying the immunological response during T. gondii infection and could have important implications for the diagnosis of acute and re-activated toxoplasmosis.
Clonality of T lymphocytes expanded with IL-2 from rheumatoid arthritis peripheral blood, synovial fluid and synovial membrane
Authors: CANTAGREL A , ALAM A , COPPIN H L , MAZlERES B , De PREVAL C
Published: July 2010

The association of rheumatoid arthritis (RA) with particular MHC class II genes suggests that autoantigen-spccific T cell clones present in joints could be central to the pathogenesis of the disease. Previous investigations on the clonal diversity of T cells infiltrating the rheumatoid synovial membrane have yielded conflicting results. With the use of Southern blot analysis, we investigated the clonalily of rheumatoid T cell lines expanded from peripheral blood, synovial fluid and synovial tissue. From peripheral blood lymphocyte (PBL) of RA patients and healthy normal controls, we also checked the consequences of two different culture conditions on the clonality of these cell lines. From control PBL, we found that in vitro non-specific expansion of non-clonal T cell populations does not create artefactual clonal selection. However, growing T cells in vitro with IL-2 seems to be able to lead to preferential expansion of cells bearing IL-2 receptor (IL-2R). We identified such in vivo activated IL-2-sensitive T cell clones frequently in RA synovial tissue (8/13) and more rarely in synovial fluid and peripheral blood (3/12). One patient presents the same T cell receptor gene rearrangements in synovial membrane of two affected joints. In RA synovial tissue, the frequency of these IL-2-responsive T cells is most prevalent among actively inflamed membranes removed early in the disease process. The role and the relevance to the disease of these I L-2-responsivc T cells remain to be elucidated.
Cell-mediated transfer of collagen-induced arthritis in mice and its application to the analysis of the inhibitory effects of interferon-gamma and cyclophosphamide
Authors: NAKAJIMA H , HIYAMA Y , TAKAMORI H , TSUKADA W
Published: July 2010

We developed a convenient and reliable procedure for the cell-mediated passive transfer of type II collagen (Cll)-induccd arthritis (CA). Spleen cells from DBA/1 mice with CA were intravenously transferred into syngeneic recipient mice. Arthritis developed only in those recipients which had received whole-body x-irradiation (8 Gy) just before cell iransfer and intrapcritoneally given soluble CII without adjuvant immediately after transfer. Non-immunized splenocytes could not induce arthritis even in irradiated recipients given soluble CII. Development of arthritis depended on the number of cells transferred; 5 ± 107 cells induced severe and long-lasting arthritis in every recipient approximately 10 days after transfer. Severity of this arthritis was clinically and histologically similar to classical CA in donors. Arthritogenic splenocytes were generated in donors no later than 20 days after priming with CII in Freund's complete adjuvant, when arthritis had yet to occur, and were detected for more than 5 weeks. One splenocyte population responsible for transferring arthritis was CD4+ T cells. We then applied this system to show that prophylactic treatment of CII-immunized mice with cyclophosphamide (CY. 7 mg/kg), but not interferon-gamma (IFN-γ. 105 U/mouse), suppressed the arthritogenic ability of their spleen cells, although both treatments inhibited the development of CA. Treatment of recipients with IFN-γ, however, inhibited the development of arthritis upon transfer with CII-immunized splenocytes. These results indicate that CY and IFN-γ act at the induction and effector phases of arthritogenic lymphocytes, respectively. Thus, this system facilitates investigation of pathological mechanisms of CA, and of mechanisms of anti-arthritics.
Comparison between the protective effects of mycobacterial 65-kD heat shock protein and ovomucoid in pristane-induced arthritis: relationship with agalactosyl IgG
Authors: GHORAISHIAN M , ELSON C J , THOMPSON S J
Published: July 2010

The IgG of patients with rheumatoid arthritis and mice with pristane induced arthritis (PIA) lends to lack the terminal galactose normally on the conserved N-acetylglucosamine linked β1–2 to mannose in IgG. The terminal N-acetylglucosamine (GlcNAc) residues of oligosaccharides on agalactosyl IgG may be an important component of the action of these glycoforms. Here, administration of ovomucoid, a glycoprotein rich in terminal GlcNAc, before pristane injection was found to reduce the incidence of PIA. This observation is the second report of an intraperitoneally administered antigen that reduces the incidence of PIA. mycobacterial 65-kD heat shock protein (hsp65) being the first. The suppressive effect of ovomucoid was not transferred from protected to naive recipients by spleen cells at the dose tested. By contrast, transfer of spleen cells from hsp65-protected mice to naive recipients conferred protection and this protection may be antibody-mediated. It is considered that ovomucoid and hsp65 protect against the development of PIA by different mechanisms.
Critical role of peripheral blood phagocytes and the involvement of complement in tumour necrosis factor enhancement of passive collagen-arthritis
Authors: FAVA R A , GATES C , TOWNES A S
Published: July 2010

Studies have implicated tumour necrosis factor-alpha (TNF-α) in type-II collagen (CII)-induced arthritis (CIA), a well established animal model of human rheumatoid arthritis. Precisely how TNF is involved in CIA is not yet clear. In this study the effects of TNF on CIA were examined, independent of its potential effects on the immune response, by performing peri-articular injection of TNF in combination with passive immunization of rats. A sub-arthritic dose (5 mg) of affinity-purified anti-CII IgG, which alone was insufficient to induce spontaneous clinical arthritis, was used throughout the study. Obvious clinical arthritis that persisted for several days was rapidly induced by injections of 100 ng TNF into hindpaws of rats that were passively immunized shortly before the TNF injection. Injections of TNF in non-immunized control rats did not induce clinical arthritis, nor did buffer-only injections in passively immunized controls. The clinical arthritic response was a local phenomenon, limited only to the TNF-injected hindpaws. No swelling was observed in the opposite, buffer-injected hindpaws, indicating the effects of TNF were not systemic. Depletion of peripheral blood phagocytes with anti-rat neutrophil antiserum before passive immunization completely abolished the ability of TNF to induce clinical arthritis, identifying phagocytic cells as the essential target cells in evoking this arthritic response. A role forcomplement activation was also demonstrated in this model through the use of a soluble recombinant version of CD35, the cell surface complement receptor type-1 (sCRl, BR155730), which significantly reduced TNF-induced arthritis in phagocyte-replete rats.
Activation of human complement by mouse and mouse/human chimeric monoclonal antibodies
Authors: SEINO J , EVELEIGH Ph , WARNAAR S , van HAARLEM L J M , van ES L A , DAHA M R
Published: July 2010

The complement (C)-activating capabilities in human serum of 32 mouse and 10 mouse/human chimeric MoAbs of different isotypes, and their fragments, were tested in vitro. Activation of C via the classical pathway (CP) was performed in 1% factor D-deficient serum in gelatin containing Veronal buffer in the presence of calcium and magnesium (GVB++), while activation of the alternative pathway of C (AP) was assessed in 10% Clq-depleted serum in the presence of 5 mm MgCl2 in GVB++. The C-activating ability of MoAbs was expressed relative to the degree of activation of complement by aggregated IgG for the CP and relative to mouse IgG 1 for the AP. All of seven mouse IgG2a MoAbs were potent activators of the CP. The results of CP activation by IgG1, IgG2b and IgG3 isotypes were different for individual MoAbs. Only three (two IgG 1 and one IgG3) of 32 mouse MoAbs were potent activators of the AP. IgG2a and IgG2b were relatively poor AP activators. There were a few MoAbs which activated both the AP and CP. Of 10 chimeric MoAbs, two IgG1, one IgG2 and one IgG4 were poor or non-activators of the CP. On the other hand, IgG2 and IgG4 were good AP activators. IgG3 was the most potent AP activator. Most of the F(ab')2 fragments were activators of the AP and displayed no activation of the CP. Fc fragments only activated the CP. whereas Fab'did not activate the CP or the AP. These studies suggest that the route of complement activation by class and subclass MoAbs can not always be predicted in advance and based only on their subclass identity.
Localization of tumour necrosis factor-alpha (TNF-α) and its receptors in normal and psoriatic skin: epidermal cells express the 55-kD but not the 75-kD TNF receptor
Authors: KRISTENSEN M , CHU C Q , EEDY D J , FELDMANN M , BRENNAN F M , BREATHNACH S M
Published: July 2010

The distribution of TNF-α, p55 TNF receptor (TNF-R) and p75 TNF-R in normal skin and uninvolved and lesional skin from psoriasis patients has been investigated, using specific mono- and polyclonal antibodies. In normal skin, and uninvolved and lesional skin from psoriasis patients, p55 TNF-R is associated with epidermal keratinocytes and a network of upper dermal dendritic cells. This suggests that the actions of TNF-a on epidermal cells in vivo are mediated by binding to the p55 TNF-R. In lesional psoriasis skin, there was staining of the parakeratotic stratum corncum and increased expression of p55 TNF-R in association with upper dermal blood vessels. Staining for p75 TNF-R in normal skin was restricted to eccrine sweat ducts and dermal dendritic cells, and was absent from the epidermis. In lesional psoriasis skin, there was staining for p75 TNF-R in association with upper dermal blood vessels and perivascular infiltrating cells. TNF-α in normal skin was predominantly localized to the basal cell layers of the epidermis, and was seen in association with eccrine ducts and sebaceous glands. In lesional psoriasis skin, and to a lesser extent in uninvolved psoriasis skin. TNF-α was distributed throughout the epidermis, and was also specifically localized to upper dermal blood vessels. Up-regulation of TNF-α. p55 TNF-R and p75 TNF-R on dermal blood vessels in psoriasis may play an important role in the pathogenesis of this condition by promoting cutaneous recruitment of inflammatory cells.
Prognostic value of phenotypic alterations in blood lymphocyte subsets in a murine retrovirus-induced immunodeficiency syndrome (MAIDS)
Authors: CHAU F , LEVACHER-CLERGEOT M , DESFORGES B , RICATTE L , SINET M
Published: July 2010

Mice infected with the Duplan strain of murine leukaemia virus (Dup MuLV), a retrovirus, develop a syndrome sharing several features with AIDS, including lymphadenopathy and profound immunodeficiency. We measured the changes in peripheral blood lymphocyte populations and evaluated their predictive value for the outcome of disease in C57B1/6 mice. Animals were inoculated with Dup MuLV (SCI /Dup MuLV confluent fibroblast supernatant or spleen extract from an infected mouse). Peripheral blood lymphocyte subsets were sequentially monitored for 73 days using flow cytometric analysis and MoAbs directly conjugated to fluorochromes. A striking fall in the Thy 1.2 cell count occurred in diseased animals, mostly affecting the CD8+ cell compartment. At the same time, the percentage of Ly5+ cells was increased. Mice were killed at day 73 and spleen and lymph node lymphocytes were analysed. Phenotypic lymphocyte modifications in peripheral blood were closely related to those in the spleen or lymph nodes. Analysis of Ly6c antigen expression on CD4+ and CD8+ cells showed a selective expansion of the CD8+ Ly6c+ subset, which may reflect a state of immune activation. Our results suggest that phenotypic alterations of peripheral blood lymphocytes are a good marker of disease progression in this model and could be a useful criterion to evaluate antiretroviral therapy.
Interactions of anti-DNA antibodies with Z-DNA
Authors: KRISHNA P , FRITZLER M J , Van de SANDE J H
Published: July 2010

Systemic lupus erythematosus (SLE) sera, two classes of serum lipoproteins, and IgG antibodies from SLE and normal sera were tested for their reactivity with a Z-DNA polymer, Br-poly (dG-dC). In all cases preferential binding to Z-DNA over B-DNA was observed. This interaction, for the most part, could be inhibited by the negatively charged phospholipid, cardiolipin, which suggests that most of the anti-Z-DNA activity associated with sera arises from relatively non-specific ionic interactions between proteins and polyanionic molecules. An assay has been described that can eliminate proteins cross-reactive with negatively charged phospholipids.
Lower antibody response to tetanus toxoid associated with higher auto-anti-idiotypic antibody in old compared with young humans
Authors: ARREAZA E E , GIBBONS J J , SISKIND G W , WEKSLER M E
Published: July 2010

The production of anti-tetanus toxoid (TT) antibody and anti-anti-TT (auto-anti-Id) antibody has been measured in three young and three old persons. Both antibodies were measured using ELISA. We found that: (i) the anti-TT response of old subjects was lower than that of young subjects, (ii) scrum auto-anti-Id antibody concentration was higher in old compared with young humans before boosting with TT, and (iii) anti-TT antibodies from different humans shared Id cross-reactivity when tested with a rabbit anti-Id antibody. Thus, the serum anti-TT antibody response to boosting was inversely correlated with the serum level of auto-anti-Id. This conclusion is consistent with the view that the higher level of auto-anti-antibody in older subjects contributes to their impaired antibody response to TT.
Polynucleotide specificity of anti-reactive oxygen species (ROS) DNA antibodies
Authors: ARA J , ALI R
Published: July 2010

Hydrogen peroxide in the presence of short wavelength UV light was able to induce alterations in native DNA fragments of 300 bp (ROS-DNA). thereby rendering it immunogenic in experimental animals. The specificity of induced antibodies was investigated by direct binding and competition ELISA. Inhibition studies revealed nearly 89% inhibition in the antibody binding by the immunogen and recognition of native B-, A- and allied conformations presented by various synthetic polynucleotides. Gel retardation assay reiterated the formation of immune complexes between induced antibodies and native and ROS-DNA fragments. It was observed that naturally occurring anti-DNA autoantibodies from systemic lupus erythemalosus(SLE) sera recognize ROS-DNA. The comparison of the specificities of anti-DNA auloantibodies from 10 SLE patients showed a 20–50-fold preference for ROS-DNA over native DNA. These results demonstrate that anti-DNA antibodies can be induced by ROS-DNA, and that some of the autoimmune DNA binding antibodies found in SLE may result from response to reactive oxygen species.
Modulation of α1-acid glycoprotein (AGP) gene induction following honey bee venom administration to adjuvant arthritic (AA) rats; possible role of AGP on AA development
Authors: YIANGOU M , KONIDARIS C , VICTORATOS P , HADJIPHTROU-KOUROUNAKIS L
Published: July 2010

Honey bee venom (HBV) administration to adjuvant arthritic (AA) rats resulted in a significant suppression of arthritis and in suppression of the hepatic acute phase α1-acid glycoprotein (AGP) gene induction at the early stages of disease development. AGP administration in AA rats resulted in acceleration of arthritis development and in increase of severity and duration of the disease. IL-1, IL-6. tumour necrosis factor (TNF) and glucocorticoids alone are not responsible for thc HBV-mediated AGP gene down-regulation. These results indicate that AGP gene expression in AA and HBV-treated AA rats involves the interaction of several factors, and that AGP plays a role for AA development in rats.
Circulating monocytes are activated in newly diagnosed type 1 diabetes mellitus patients
Authors: JOSEFSEN K , NIELSEN H , LORENTZEN S , DAMSBO P , BUSCHARD K
Published: July 2010

Investigations in the BB rat and the non-obese diabetic (NOD) mouse have provided substantial evidence for the involvement of the monocyte/macrophage system in the development of type 1 diabetes mellitus. However, it is not known whether monocytes play the same role in the pathogenesis of human type 1 diabetes. We investigated this problem in a longitudinal study of 29 recent-onset type 1 diabetes mellitus patients. Monocyte chemotaxis, phagocytosis and superoxide production as well as metabolic and haematological parameters were studied immediately after diagnosis and 6 months later. At diagnosis the patients had activated casein and C5a chemotaxis (casein 70 ± 9 versus 150 ± 5 (mean ± s.e.m.), P < 0001; C5a 137 ± 10 versus 158 ±5, P < 005 (activation immobilizes monocytes, reducing the measured values)), and activated superoxide production (3·6 ± 03 versus 3·0 ± 03, P < 0·05), After 6 months casein chemotaxis (115 ±16 versus 150 ±5, P < 005) and Candida phagocytosis (33 ±01 versus 2·8 ±0·2. P < 0.001) were still activated. There was no correlation with other clinical or paraclinical parameters. We conclude that the circulating monocytes in newly diagnosed type 1 diabetes patients are activated. It is reasonable to expect that monocytes at the local site of inflammation in pancreas are even further activated. This could play a pathogenic role in β cell destruction.
GM-CSF, IL-1α, IL-β, IL-6, IL-8, IL-10, ICAM-1 and VCAM-1 gene expression and cytokine production in human duodenal fibroblasts stimulated with lipopolysaccharide, IL-1α and TNF-α
Authors: PANG G , COUCH L , BATEY R , CLANCY R , CRIPPS A
Published: July 2010

The role of mucosal fibroblasts in intestinal inflammatory reactions is not known. In this study, we demonstrate that fibroblasts grown from histologically normal human duodenal biopsy tissues expressed mRNA genes for granulocyte-macrophage colony-stimulating factor (GM-CSF). IL-lα, IL-1β, IL-6, IL-8, IL-10, intercellular adhesion molecule-I (ICAM-I) and vascular cell adhesion molecule-1 (VCAM-i) when stimulated with lipopolysaccharide (LPS) or IL-1α. The increased mRNA expression of GM-CSF, IL-1α IL-1β IL-6 and IL-α in response to IL-1α and LPS stimulation was time- and dose-dependent. In contrast. IL-10 was weakly expressed when fibroblasts were stimulated with LPS. IL-1α or tumour necrosis factor-alpha (TNF-α), but the expression was enhanced in the presence of cycloheximide combined with optimal concentrations of LPS. IL-1α or TNF-α. IL-1α was a more potent stimulator than LPS for GM-CSF. IL-6, IL-8 and I L-10 expression, but not for IL-1α and IL-1β. Increased GM-CSF. lL-6 and IL-8 gene expression was associated with the production of cytokine proteins in culture supernatant, but IL-1α and IL-1bL remained undetectable. Dexamethasone suppressed both gene expression and protein production of GM-CSF. IL-6 and IL-8 when fibroblasts were exposed to IL-1α. TNF-α stimulated the release of GM-CSF. IL-6 and IL-8 and, combined with IL-1α. cytokine production was enhanced synergistically. Finally, both LPS and IL-1ã up-regulated ICAM-I and VCAM-1 gene expression. These findings implicate duodenal fibroblasts in the initiation and/or regulation of intestinal inflammation.
Attenuation oflpr-graft-versus-host disease (GVHD) in MRL./lprspleen cell-injected SCID mice byin vivotreatment with anti-Vβ8.1,2 monoclonal antibody
Authors: HOSAKA N , NAGATA N , MIYASHIMA S , IKEHARA S
Published: July 2010

When MKL/lpr (H-2k) spleen cells were intraperitoneally injected into C.B-17-scid/scid (severe combined immunodeficient (SCID)) (H-2d) mice, the SCID (SCID-MRL/lpr) mice manifested a severe wasting syndrome with weight loss, splenic atrophy, and lymphoid cell infiltration in the liver and lung, as seen in lpr-GVHD. In contrast. MRL/+ spleen cell-injected SCID (SCID-MRL/+) mice did not show Ipr-GVHD. The spleens of SCID-MRL/lpr mice showed progressive increases in donor CD4+ and CD8+ T cells from 4 to 12 weeks after injection and a decrease in B cells at 12 weeks. SCID-MRL/+ mice showed a stable engraftment of CD4+ and CD8+ T cells and a progressive increase in B cells. Analyses of T cell receptor (TCR) repertoires (Vβ6, Vβ8.1,2 and Vβ11) revealed that the Vβ8.1,2+ T cells were found more frequently in SCID-MRL/lpr mice than in SCID-MRL/+ mice. When SCID-MRL/lpr mice were treated with intraperitoneal injection of an anti-Vβ8.1,2+ (KJ16) MoAb, Vβ8.1,2+ T cells were markedly depleted, and the severity of/pr-GVHD was attenuated at 4 and 8 weeks after treatment, in contrast to normal rat IgG-injected SCID+MRL/lpr mice. However, the KJI6 MoAb-treated SCID-MRL/lpr mice suffered from severe lpr-GVHD 12 weeks after treatment, although Vβ8.1,2+ T cells were still maintained at a low level. These findings suggest that Vβ8.1,2+ T cells are a major T cell population (hat mediates lpr-GVHD in the early stage of lpr-GVHD. but that in the later stage, the other T cell populations may proliferate naturally or in accordance with the depletion of Vβ8.1,2+ T cells, and contribute to the development of lpr-GVHD.
IgG-mediated phagocytosis in regenerated splenic tissue
Authors: CLAYER M T R , DREW P A , LEONG A S-Y , JAMIESON G G
Published: July 2010

The risk of severe infections after splenectomy is well established. Operations such as auto-transplantation, splenic artery ligation or partial resection have been advocated for the retention or regeneration of splenic tissue following splenic trauma. The potential of such tissue to protect from infection is unclear. The ability of splenic tissue to phagocytose IgG opsonized syngeneic erythrocytes was measured in rats 6 months following splenectomy and splenic auto-transplantation, splenic artery ligation, total or partial splenectomy, and compared with eusplenic controls. In eusplenic and partially splenectomized rats 71% of the label was cleared at 3 h, compared with approximately 50% in rats following total splenectomy, splenectomy and splenic autotransplantation or splenic artery ligalion. The autotransplanted and the ligated splenic tissue cleared less than 10% compared with control spleen, but there was no difference between them when clearance was expressed as uptake per gram of tissue. Splenic autotransplants and ligated spleens were small and histologically abnormal, with an increase in the red pulp, significantly less white pulp and marginal zone, and the frequent absence of a central arteriole in the white pulp. The clearance of label was proportional to the amount of red pulp in the tissue, although the red pulp from the regenerated tissues was not as efficient at phagocytosis as control red pulp. The tissue which regenerated following autotransplantation or splenic artery ligation did not result in greater clearance of erythrocytes from the circulation than that which occurred in splenectomized rats.
Functional heterogeneity of mast cells isolated from different microenvironments within nasal polyp tissue
Authors: FINOTTO S , DOLOVICH J , DENBURG J A , JORDANA M , MARSHALL J S
Published: July 2010

Nasal polyposis is a chronic inflammatory condition of the upper airways characterized by infiltration of activated inflammatory cells, including mast cells, both in the epithelium and in the stroma. The aim of this work was to study human mast cells derived from two different anatomical sites within the same nasal polyp tissue. To this end, we isolated two distinct mast cell populations, one from the epithelial and the other from stromal layers of individual human nasal polyp tissues. We examined the mediator content of the two mast cell populations and found that stromal mast cells had a significantly higher content of tryptase compared with the epithelial mast cells from the same tissue. In addition, mast cells from the stromal compartment, but not those from the epithelium, released a significant amount of histamine after anti-IgE stimulation. By contrast, both populations released over 50% of the total histamine after non-specific stimuli (A231K7 10-6 M). The content of mediators and the response to immunological activation were not significantly altered in patients receiving topical steroid therapy. It remains to be determined if the observed differences are the result of an intrinsic characteristic of the mast cell populations localized to separate tissue compartments, or reflect a different in vivo exposure to stimuli such as antigens, or different surrounding structural or infiltrating cells. In conclusion, these data provide evidence of functional heterogeneity und differences in mediator content between mast cell subpopulations from a single human tissue. The failure of release of epithelial mast cell mediators from an immunologic stimulus may have implications concerning acute effects of antigen exposure in nasal polyposis.
High numbers of autoantigen-reactive mononuclear cells expressing interferon-gamma (IFN-γ), IL-4 and transforming growth factor-beta (TGF-β) are present in cord blood
Authors: YU M , FREDRIKSON S , LINK J , LINK H
Published: July 2010

Umbilical cord blood of neonates and peripheral blood of healthy adults were analysed by in situ hybridization for numbers of mononuclear cells (MNC) expressing the cytokines IFN-γ, TGF-β and IL-4 mRNA without culture and after culture in the presence of acetylcholine receptor (AChR), myelin basic protein (MBP) and peripheral myelin protein P2. These antigens were chosen since they represent autoantigens in putatively immune-mediated neurological diseases, The numbers of cells expressing cytokine mRNA after 72 h culture in the presence of AChR, MBP and P2 were higher in cord blood than in peripheral blood of healthy adults. IFN-γ, TGF-β and IL-4 were always elevated in parallel. In cord blood there was a pronounced reactivity to several of the tested antigens, while such broad reactivity was not found in peripheral blood of healthy adults. No differences in cytokine mRNA expression were found between cord blood and peripheral blood of adults when cells were analysed without culture. The results show a capacity of cord blood cells to react to several autoantigens by the up-regulation of cytokine mRNA expression.
Antibody-directed complement-mediated cytotoxicity to hepatocytes from patients with chronic hepatitis B
Authors: MICHALAK T I , LAU J Y N , McFARLANE B M , ALEXANDER G J M , EDDLESTON A L W F , WILLIAMS R
Published: July 2010

The susceptibility of hepatocytes from patients with chronic hepatitis B to complement-dependent cytotoxicity mediated by heterologous antibodies to hepatitis B virus core (anti-HBc) and surface (anti-HBc) antigens and to hepatic asialoglycoprotein receptor was examined using a micro-cytotoxicity assay. The anti-HBc-induced cytotoxicity was found to be markedly enhanced against hepatocytes isolated from patients with chronic active hepatitis (72·6 ± 9·5% (mean±s.e.m.); n= 6) over that against hepatocytes from individuals with chronic persistent hepatitis or inactive liver cirrhosis (40·6 ± 18·6%; n= 4) (P= 0·019). Overall, values of the anti-HBc-directed cytotoxicity were higher in patients positive for HBcAg in hepatocytes and seropositive for hepatitis B virus e antigen (HBeAg), Hepatocytotoxicity was also exerted by anti-HBs and anti-asialoglycoprotein receptor antibodies in the presence of complement, but it was not seemingly related to disease activity. These results indicate that hepatitis B virus core and surface antigens and asialoglycoprotein receptor at the hepatocyte surface can be recognized by antibodies, and raise the possibility that complement-dependent cytolysis may contribute to the injury of hepatitis B virus-infected hepatocytes. The data also suggest that liver cells of patients with severe chronic hepatitis might be more susceptible to anti-HBc antibody-directed complement-mediated cytotoxicity than those with inactive liver histology.