bluuebunny/crossref_metadata_2025 huggingface dataset:
T cell interactions in active rheumatoid arthritis: insights from the human autologous mixed lymphocyte reaction as a model of T cell activation cascade
Authors: SAKANE T , MURAKAWA Y , TAKENO M , SHIGEKI T , NAGAFUCHI H , MIKI T
Published: July 2010

The autologous mixed lymphocyte reaction (AMLR) represents the activation, proliferation and differentiation of T cells in response to signals from autologous non-T cells. Using monoclonal anti-Leu8 antibody to isolate subpopulations of human CD4+ and CD8+ T cells, we have investigated the role of these subpopulations in the T cell activation cascade during the course of AMLR. In normal subjects, CD4+ Leu8+ cells are necessary for the initiation of the AMLR response, and sequentially lead to activation and proliferation of both CD4+ Leu8- cells and CD8+ Lcu8+ cells. The activated CD8+Lcu8+ cells, in turn, induce CD8+ Leu8- cells to generate proliferation of the latter cells. Soluble mediators could be involved in the T cell activation cascade induced by the AMLR. Patients with active rheumatoid arthritis have a profound defect in the AMLR. Further analysis indicates that rheumatoid arthritis CD8+ T cells are markedly defective as responding cells in the AMLR. The impaired AMLR response by CD8+ cells cannot be reconstituted with AMLR-derived supernatants from normal T cells. The data suggest that the defective CD8+ T cell function may contribute to the pathogenesis of the disease.
Detection of circulating FcɛR2/CD23+ monocytes in patients with rheumatic diseases
Authors: BECKER H , POTYKA P , WEBER C , FEDERLIN K
Published: July 2010

Recently, in vitro studies have demonstrated that expression of FcɛR2/CD23 on normal monocytes can be specifically induced by IL-4. In order to investigate the interaction of IL-4 and monocytes in rheumatic diseases, flow cytometry studies were performed. Elevated numbers of circulating FcɛR2/ CD23+ monocytes were detected in patients with progressive systemic sclerosis (PSS) as compared with controls, in addition, supernatants derived from phytohaemagglutinin-stimulated peripheral blood mononuclear cells of PSS patients contained high activity to induce FcɛR2/CD23 on CDI4+ monocytes. An increased frequency of FcɛR2/CD23+ monocytes was also observed in rheumatoid arthritis, and sequential studies in patients with systemic lupus erythematosus showed a close relationship between FcɛR2/CD23+ monocytes and disease activity. It is suggested that IL-4 has an important role in the pathogenesis of PSS by activating monocytes, and might also contribute to monocyte activation in other rheumatic diseases.
Failure Co detect deposition of complement and immunoglobulin in allergen-induced late-phase skin reaction in atopic subjects
Authors: FREW A J , KAY A B
Published: July 2010

There still remains some controversy regarding the possible role of immune complexes in the pathogenesis of the late-phase skin reaction (LPSR). To assess this, skin biopsies were obtained from LPSR induced in atopic human subjects 6, 24 and 48 h after allergen challenge. Cryostat sections were stained by direct immunofluorescence for the presence of fibrinogen, immunoglobulin classes IgM and IgG and for the the complement components C1q and C3c. Complement components were observed in only two of the 29 biopsies studied. In both instances, only C3c was detected. One of these subjects also had unequivocal IgG staining at 6 h. IgM staining was detected in two out of 10 subjects at 6 h but no significant deposition of immunoglobulins could be found at 24 or 48 h. Fibrinogcn deposition was observed in about half of the biopsies at each time-point. This study suggests that substantial complement and immunoglobulin deposition arc not overt features of the allergen-induced LPSR. although the presence of small amounts of immune complexes, below the sensitivity of the method employed cannot be excluded. Fibrin deposition occurs in the LPSR but docs not appear to be a prerequisite for LPSR development.
Analysis of the cells involved in the lymphoproliferative response toCoxiella burnetiiantigens
Authors: IZZO A A , MARMION B P , HACKSTADT T
Published: July 2010

Vaccination with an inactivated, whole cell, Q fever vaccine (Q-vax) induces lasting antibody conversion and a positive delayed-type hypersensitivity (DTH) skin reaction in about 60% of recipients but a long-lasting positive lymphoproliferative or mitogenic response to C. burnetii antigens with peripheral blood mononuclear cells (PBMC) in 85–95% of subjects. Analysis of the lymphoproliferative response to C. burnetii antigens has now been made by fractionationreconstitution experiments with PBMC from vaccinees, from past infections, and from healthy controls. The major contributor to the response in immune subjects proved to be the T lymphocyte. T cells were stimulated by both the phase I and phase II antigens of two prototype strains of C. burnetii and responses were greatly amplified by addition of IL-2. Similar T lymphocyte stimulation profiles were obtained with the ‘Priscilla’ strain of C. burnetii which represents a different biotype of Coxiella isolated from Q fever endocarditis; Q-vax is therefore likely to protect against endocarditis strains. Fractionation-reconstitution experiments with T and B cells from vaccinees and subjects infected in the past, using various antigenic or haptenic fractions from C. hurnetii indicate that protein, non-lipopolysaccharide components of the organism are responsible for the mitogenic response of immune T cells. However, the role of the lipopolysaccharide in the protective immunogen has still to be defined.
Examination of serum class I antigen in liver-transplanted rats
Authors: SUMIMOTO R , SHINOMIYA T
Published: July 2010

We examined the appearance of donor (DA) type class I antigen in the serum of rats that had received isogeneic (DA→DA) or allogencic (DA→PVG, DA→BN, DA→LEW) liver transplants with or without cyclosporin A treatment, using two-site enzyme immunoassay. We also tested the serum titre of class I antigen in the normal DA rats with either 70% hepatectomy or cyclosporin A treatment, in order (o clarify the relationship between the fluctuation in the scrum titre of class I antigen in the recipient and the outcome of the transplanted liver graft. The suppression of liver graft rejection by cyclosporin A treatment significantly lowered the serum level of donor liver-derived class I antigen as compared with that of the recipient without cyclosporin A for a certain period. However, there was almost no correlation between the intensity of rejection of the liver graft and the serum level type class I among these allogencic rejection and non-rejection liver transplantation combinations. The amount of donor-type class I antigen in the recipient’s scrum is dependent on whether the grafted liver is severely damaged following partial hepateetomy, whether the liver has associated biliary complications or ischaemic damage, or whether the liver has had absolute residual parenchymal cell volume or function following liver rejection. Our results suggest that the appearance of donor type class I anligen following liver transplantation is dependent on many factors, and therefore the litre of serum class I antigen may not always be a decisive indicator of liver graft rejection.
Effects of granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-2, interferon-gamma (IFN-γ), tumour necrosis factor-alpha (TNF-α) and IL-6 on the production of immunoreactive IL-1 and TNF-α by human monocytes
Authors: DANIS V A , FRANIC G M , RATHJEN D A , BROOKS P M
Published: July 2010

The effects of GM-CSF, IL-2, IFN-γ, TNF-α and IL-6 on the production of IL-1 (both secreted and cell associated) and TNF-α by peripheral blood monocytes were studied. Monocytes were cultured for 20 h in suspension and in serum-free conditions which minimized background stimulation of monokine production. GM-CSF, IL-2 and TNF-α directly induced the production of cell-associated IL-1 but little or no IL-1 or TNF-α secretion. Combination of GM-CSF with IFN-γ, IL-2 or TNF-α synergistically enhanced IL-1 secretion and had an additive effect on cell-associated IL-1 production. Combination of IL-2 with IFN-γ or TNF-α also synergistically enhanced IL-1 secretion but the effect on cell-associated IL-1 production was less than additive. GM-CSF synergisticaily enhanced TNF-α secretion induced by IFN-γ but not by tipopolysaccharide. GM-CSF did not enhance TNF-α secretion induced by IL-2 or TNF-α. In contrast, IL-2 synergistically enhanced TNF-α secretion induced by IFN-γ. These results are discussed in relation to cytokine involvement in rheumatoid arthritis.
Defective neutrophil and lymphocyte function in leucocyte adhesion deficiency
Authors: LAU Y L , LOW L C K , JONES B M , LAWTON J W M
Published: July 2010

We report a Chinese girl with the moderate phenotype of leucocyte adhesion deficiency (LAD), presenting with persistent omphalitis and recurrent soft tissue infections. She had subnormal adhesion-dependent neutrophil functions, such as chemotaxis and chemiluminescence response to a particulate stimulant (opsonised zymosan). Despite her adequate humoral response to documented herpes simplex virus type 1, parainfluenza type 2 and adenovirus infection in vivo. there was marked impairment in the generation of plaque-forming cells (PFC) driven by pokeweed mitogen (PWM) in vitro. IgM PFC were less severely affected than IgG and IgA PFC, probably because IgM production is less dependent on T cell help than IgA and IgG production. The patient’s B cells and accessory cells had reduced function compared with the control subsets, while helper function of her CD4+ cells was virtually absent in the PWM-driven PFC assay. She also had marked defect in natural killer cell activity. The proliferation of her lymphocytes was normal to several plant Iectins, including phytohaemagglutinin, concanavalin A and PWM, but markedly defective to OKT3.
Sequential serological studies of homosexual men with and without HIV infection. Epstein-Barr virus activation preceding and following HIV seroconversion
Authors: SCHATTNER A , HANUKA N , SAROV B , SAROV I , HANDZEL Z , BENTWICH Z
Published: July 2010

Viral cofactors may be important in the pathogenesis of HIV infection and the development of AIDS, but their role is still imperfectly understood. Sequential serological studies were performed in a cohort of 100 homosexual men and 70 matched healthy controls over a mean period of 4 years. Of the patients, 18 were found to be HIV+ on admission to the study and 15 seroconverted to HIV+ during the follow up (seroconversion group). Serum antibodies of both IgG and IgA isotypes against Epstein-Barr virus (EBV) and cytomegalovirus (CMV) were determined. IgG antibodies indicate past infection, while a marked increase in IgG titre or a positive IgA titre were taken to indicate active infection or reactivated latent infection. EBV and CMV infections were about two to four times more prevalent in the homosexual men both HIV– and HIV+, compared with controls. Active infections were increased in the homosexual men and particularly in the HIV+ patients. The seroconversion group revealed activation of both EBV and CMV following HIV infection. When the antibody profile of seroconverting patients at the time preceding seroconversion was compared with a matched group of 39 homosexual men who remained HIV–, no change was found in CMV antibodies, but four out of 15 (26.6%) of the patients had high titres of anti-EBV IgA preceding seroconversion, as compared with only one out of 39 (2.6%) of HIV– homosexual men (/<0.05). This suggests a role for EBV reactivation in the pathogenesis of HIV infection in some patients.
Antibodies to endothelial cells in systemic lupus erythematosus: a potential marker for nephritis and vasculitis
Authors: D’CRUZ D P , HOUSSIAU F A , RAMIREZ G , BAGULEY E , MCCUTCHEON J , VIANNA J , HAGA H-J , SWANA G T , KHAMASHTA M A , TAYLOR J C , DAVIES D R , HUGHES G R V
Published: July 2010

Using an ELISA, anti-endothelial cell antibodies (AECA) have been found in sera obtained at the time of renal biopsy in 46 out of 57 patients (81%) with systemic lupus erythematosus (SLE) and nephritis (mean binding index (BI) = 84%± 52.8) compared with 22 out of 50 SLE patients (44%) without nephritis (mean BI = 45%±35.9). Seventy normal human sera had a mean BI of 10%±9.8. The highest levels were seen in patients with diffuse proliferative glomerulonephritis (WHO grade IV) and in patients with proteinuria and nephrotic syndrome. When the biopsies were assessed for activity and chronicity scores, AECA were associated with active renal lesions (P< 0.001). AECA levels correlated with low complement levels but not with anti-DN A antibodies to extractable nuclear antigens (ENA), anti-cardiolipin or anti-neutrophil cytoplasmic antibodies. The presence of AECA conferred a positive predictive value of 0.68 for the presence of nephritis. Twenty-five patients had active vasculitis at the time of assay and the highest AECA values were seen in patients with both nephritis and vasculitis. No correlation was seen with serum immunoglobulin levels and immune complexes did not bind significantly to the endothelial surface. The possible role of these antibodies as a marker in lupus nephritis is discussed.
Anti-IgG antibodies in rheumatic diseases cross-react withStreptococcus mutansSR antigen
Authors: ACKERMANS F , PINI A , WACHSMANN D , SCHOLLER M , OGIER J , KLEIN J-P
Published: July 2010

We have previously shown that SR protein, a S. mutans major cell wall protein, as well as the recombinant protein SR (rSR) share common epitopes with human IgG. Since this antigenic mimicry could play a role in the induction of Anti IgG, we have examined, in k-ELISA, the presence of antibodies reacting with S. mutans SR proteins and S. mutans whole cells in sera from 36 patients with rheumatic diseases. The majority of the 36 sera showed a high reactivity with rSR when compared with control sera. Eight highly positive sera were further purified on rSR and human IgG sorbents and tested against both rSR and IgG in ELISA and Western blotting. The affinity-purified antibodies reacted strongly with rSR, IgG and IgG Fab fragments but failed to react with IgG Fc fragment. In Western blotting the addition of unlabelled IgG abolished the reactivity of affinity-purified biotinylatcd antibodies with all antigens, confirming the existence of a common epitopc shared by rSR and human IgG heavy chain. We show the existence in rheumatic diseases of high litres of anti-human IgG antibodies cross-reactive with S. mutans SR proteins. Those antibodies are principally IgG and react with the Fd part of the Fab fragment. We can hypothesize from the above data that this antigenic mimicry existing between S. mutans SR-related antigens and human IgG could play a role in the synthesis of at least a part of the anti-IgG antibodies present in rheumatic diseases sera.
Bacterial lipopolysaccharide enhances deposition of immune complexes and exacerbates nephritis in BXSB lupus-prone mice
Authors: GRANHOLM N A , CAVALLO T
Published: July 2010

Systemic autoimmune disease is influenced by genetic, immunological, hormonal, and environmental factors. Although environmental factors are major agents that induce or exacerbate autoimmune diseases, the mechanism(s) and the molecular events by which they operate remain poorly understood. Here we used the lupus-prone BXSB mouse as an animal model of systemic autoimmune disease, and we used a bacterial lipopolysaccharide (LPS) as a surrogate infectious agent to gain some insight into the mechanism(s) by which infectious agents exacerbate autoimmune diseases. Our experimental protocol was designed to address three questions: (i) whether spontaneous polyclonal B cell activation (PBA) that occurs in BXSB mice could be further enhanced by bacterial LPS; (ii) whether repeated exposure to LPS would exacerbate autoimmune disease, as reflected by enhanced deposits of immune complexes (ICs) in kidneys and exacerbated nephritis; and (iii) whether the mechanism by which LPS exacerbates nephritis might involve interference with blood cell carrier function, mononuclear phagocyte function, or both. BXSB mice were injected with LPS (25 μg) twice a week for 5 weeks; control autoimmune BXSB mice and immunologically normal (C57BL/6) mice were injected with vehicle only. The three groups of mice were then challenged with soluble ICs to assess the kinetics of their disappearance from the circulation, their uptake by the mononuclear phagocyte system (liver, spleen), their distribution in target organ (kidney), and blood cell carrier function. The results indicate that: (i) spontaneous PBA can be enhanced further by LPS; (ii) exposure to LPS results in increased deposits of endogenous ICs in kidneys and exacerbated nephritis; and (iii) defective handling of ICs by the mononuclear phagocyte system and impaired blood cell carrier function are contributory factors to exacerbated nephritis, but that mechanisms in addition to passive localization of ICs may also be operative.
Effects of IL-5, granulocyte/macrophage colony-stimulating factor (GM-CSF) and IL-3 on the survival of human blood eosinophilsin vitro
Authors: TAI P-C , SUN L , SPRY C J F
Published: July 2010

The mechanisms that could affect the lifespan of eosinophils after they have left the bone marrow, and their capacity to respond to activation factors were studied by examining the effects of IL-5, GM-CSF and IL-3 on purified human blood eosinophils in culture. All three cytokines prolonged the lifespan of the majority of blood eosinophils. This effect was dose dependent: IL-5 > GM-CSF > IL-3. Light density eosinophils from most patients had a longer lifespan in culture than did normal density eosinophils, with or without the three cytokines. Eosinophil death in the absence of these cytokines occurred by apoptosis. Eosinophils from two patients did not survive when cultured with IL-5, although they survived in the presence of IL-3 or GM-CSF- IL-5, GM-CSF and IL-3 induced the expression of the activation epitope on the eosinophil ribonucleases recognized by monoclonal antibody EG2. We conclude that small amounts of IL-5, GM-CSF and IL-3 prevented programmed cell death in human blood eosinophils and induced the expression of the activation forms of eosinophil ribonucleases. We suggest that differences in the capacity of normal and light density eosinophils to survive in culture, and in the ability of eosinophils from some patients to respond to IL-5 could account for variations in the severity of disease seen in patients with persistent eosinophilia.
IgG human monoclonal anti-DNA autoantibodies from patients with systemic lupus erythematosus
Authors: WINKLER T H , JAHN S , KALDEN J R
Published: July 2010

We describe the production of six mouse-human heterohybridomas secreting human IgG anti-dsDN A antibodies derived from patients with systemic lupus erythematosus (SLE). Peripheral blood cells used for fusion experiments were from patients who were shown to have high numbers of anti-DNA secreting B cells in the peripheral blood. All monoclonal antibodies bind to dsDNA in ELISA systems, five are reactive with Crithidia lucilae kinetoplasts and three precipitate dsDNA in the Farr assay. Inhibition studies revealed a remarkable specificity for certain polynucleotide structures. To our knowledge these are the first hybridomas described in the human system that secrete anti-dsDNA antibodies of the IgG class.
Impaired tumour necrosis factor-α (TNF-α) production and abnormal B cell response to TNF-α in patients with systemic lupus erythematosus (SLE)
Authors: MITAMURA K , KANG H , TOMITA Y , HASHIMOTO H , SAWADA S , HORIE T
Published: July 2010

We examined the TNF-α activity in culture supernatants of monocytes isolated from the peripheral blood of patients with SLE and of normal individuals. The monocytes from patients with SLE stimulated with silica particles, lipopolysaccharide or Staphylococcus aureus Cowan I secreted significantly lower amounts of TNF-α than did normal monocytes. A decreased TNF mRNA expression was observed in peripheral blood mononuclear cells stimulated by mitogens from patients with SLE. Furthermore, we examined the effect of recombinant TNF-α (rTNF-α) on the B cell function in SLE patients. rTNF-α inhibited the spontaneous Bcell proliferation of SLE, but tended to enhance the normal Bcell proliferation. Spontaneous IgM production from SLE Bcells was inhibited by rTNF-α, but that from normal B cells was not. Spontaneous IgG production was unaffected by rTNF-α. Also, rTNF-α did not affect the viability of B cells. These findings suggest that an impaired TNF-α production and an abnormal B cell response to TNF-α play a role in the immunological dysfunction in patients with SLE.
Tumour necrosis factor/cachectin plays a key role in autoimmune pulmonary inflammation in lupus-prone mice
Authors: DEGUCHI Y , KISHIMOTO S
Published: July 2010

The role of tumour necrosis factor-alpha (TNF-α) in the development of autoimmune pulmonary inflammation has been investigated in lupus-prone mice. An increase in TNF-α mRNA level from whole lung preparation of lupus-prone mice was evident, from 3 weeks to 12 weeks during growing process, as shown by Northern blot analysis, but not in control mice. Furthermore, it is also found that the major source of this increase in TNF-α mRNA was attributed to infiltrating mononuclear cells found within the lung. Treatment of lupus-prone mice with rabbit anti-mouse TNF-α IgG prevented the development of pulmonary inflammation lesions such as lung fibrosis and alveolitis. These results suggest that an increased TNF-α production by infiltrating mononuclear cells in the lungs of lupus-prone mice may play a role in the development of autoimmune pulmonary inflammation and in significant changes of cytokines and the immune responses in pulmonary inflammation lesions of lupus-prone mice.
Estimation of amounts of anti-La(SS-B) antibody directed against immunodominant epitopes of the La(SS-B) autoantigen
Authors: GORDON T P , GREER M , REYNOLDS P , GUIDOLIN A , MCNEILAGE L J
Published: July 2010

The contribution of circulating anti-La(SS-B) antibody to the hypergammaglobulinaemia seen in primary Sjögren’s syndrome is unknown. In this study levels of anti-La(SS-B) antibody directed against three immunodominant epitopes of the anti-La(SS-B) autoantigen were measured by ELISA in 84 anti-La(SS-B)+ sera using purified recombinant protein and antibody affinity-purified against the three anti-La(SS-B) fusion proteins. There was marked variation in the amounts of IgG anti-La(SS-B) antibody detected, with levels ranging from 0.02 mg/ml to 11 mg/mt. The anti-La(SS-B) levels were > 1 mg/ml in 61% of patients; in 18% of sera the anti-La(SS-B) level constituted 10% or more of the total serum IgG. However, other patients were seen with marked hypergammaglobulinaemia and low anti-La(SS-B) concentrations. These results support an antigen-driven mechanism for the anti-La(SS-B) response and suggest that anti-La(SS-B) antibody production is regulated independently of other immunoglobulins.
Anti-IL-lα autoantibodies in patients with rheumatic diseases and in healthy subjects
Authors: SUZUKI H , AYABE T , KAMIMURA J , KASHIWAGI H
Published: July 2010

We have developed a quantitative assay for IgG autoantibodies against IL-lα using protein A-Sepharose CL-4B. We examined the autoantibodies in sera from 107 healthy subjects, 151 patients with rheumatoid arthritis (RA), 64 patients with systemic lupus erythematosus (SLE) and 16 patients with systemic sclerosis. The frequency of positive sera for the autoantibodies in patients with RA was 16.6%, which was about three times more frequent (P<0.01) than that in healthy subjects (5.6%) or that in patients with SLE (4.7%). Only one serum of 16 patients with systemic sclerosis was positive for the autoantibodies. Neutralizing activity of the autoantibodies was demonstrated by murine thymocyte proliferation assay. The concentrations of IgG at 50% inhibition of IL-lα (15 pM) induced thymocyte proliferation ranged between 0.1 and 0.5 mg/ml. A time-course study showed fluctuations of the titres of the autoantibodies in parallel with the disease activity of RA. These results suggest that the anti-IL- lα autoantibodies present in the sera and possibly some other body fluids may be involved in the regulation of IL-1 activity in rivo.
Impaired T cell signal transduction through CD28 in a patient with idiopathic thrombocytopenia
Authors: PÉREZ-BLAS M , ARNAIZ-VILLENA A , GÓNGORA R , SEGURADO O G , VIVANCO J L , REGUEIRO J R
Published: July 2010

We describe an infant whose peripheral blood mononuclear cells were unable to proliferate or synthesize IL-2 in response to a mitogenic combination of antibodies directed against CD2 and CD28. This peculiar defect, which has been stable to dale, was attributed to an impairment in CD28-mediated T cell activation, because further comitogenic combinations containing anti-CD28 monoclonals also failed to induce normal proliferation of the patient’s T cells. In contrast, proliferation after membrane stimulation (with anti-CD2, recombinant IL-2, or certain lectins) or transmembrane activation (with phorbol ester and calcium ionophore) was normal, suggesting that his lymphocytes did not have a general membrane or intracellular signalling impairment. A T cell line derived from the patient confirmed the existence of a severe defect in CD28-mediatcd T cell proliferation, but also showed a profound impairment in CD3-induced T cell proliferation. Other cell surface molecules like CD2 and CD25 were, in contrast, capable of transducing normal proliferation signals. As all relevant molecules were detectable by cytofluorography and immunoprecipilalion, we conclude that the patient’s lymphocytes had an intrinsic defect in the delivery of CD28-mediated signals which, in the absence of monocytes, also affected CD3-mediated proliferation. The study of this novel kind of immunodeficiency may help to unravel the complex interactions that take place among CD2. CD3 and CD28 during T cell activation. The presence of an idiopathic thrombocytopenia in the patient suggests the intriguing possibility of a role for CD28 in the maintenance of peripheral blood platelets levels, although alternative interpretations are not ruled out.
Reduced frequency of nickel allergy upon oral nickel contact at an early age
Authors: VAN HOOGSTRATEN I M W , ANDERSEN K E , VON BLOMBERG B M E , BODEN D , BRUYNZEEL D P , BURROWS D , CAMARASA J G , DOOMS-GOOSSENS A , KRAAL G , LAHTI A , MENNE T , RYCROFT R J G , SHAW S , TODD D , VREEBURG K J J , WILKINSON J D , SCHEPER R J
Published: July 2010

From animal studies we know that oral administration of T-dependent antigens before sensitization effectively induces systemic immune unresponsiveness. Such ‘oral tolerance’ is persistent, dose-dependent, antigen-specific and presumably T suppressor cell-mediated. Oral tolerance induction could be an effective way to prevent undesired T cell-mediated immune functions, such as playing a role in allograft reaction, autoimmune and allergic diseases. In the present study allergic contact hypersensitivity (ACH) to nickel, currently presenting the most frequent contact allergy in man, was chosen to establish the feasibility of oral prevention of undesired T cell-mediated immunity in man. Potentially tolerizing (oral nickel contacts via orthodontic braces) as well as sensitizing (ear piercing) events were studied retrospectively in 2176 patients attending nine European patch test clinics. Patients were interviewed by means of a confidential questionnaire. The results show that ear piercing strongly favoured development of nickel ACH. More importantly, patients having had oral contacts with nickel-releasing appliances (dental braces) at an early age, but only if prior to ear piercing, showed a reduced frequency of nickel hypersensitivity. Frequencies of other hypersensitivities, in particular to fragrance, were not affected. These results support our view that induction of specific systemic immunologic tolerance by timely oral administration of antigens is feasible in man.
Circulatory antigen of Heymann nephritis. III. Presence of the 70-kD circulatory protein in the immune deposits of Heymann nephritis
Authors: SINGH A K , SCHWARTZ M M
Published: July 2010

An antigen of 70 kD size has been isolated previously from normal rat serum which has immunological cross-reactivity to the Heymann nephritis antigen, F x 1 A. Its role in the pathogenesis of Heymann’s nephritis was unknown. In this investigation we tested for the presence of 70-kD circulatory antigen in the glomerular immune deposits of Heymann’s nephritis. Further, its presence was correlated with severity of disease. It was observed that the presence of the 70-kD antigen strongly correlated with the existence of electron-dense deposits in the lamina rara externa (LRE) of the glomerular capillary wall and with pathologic proteinuria. Temporally, the presence of the 70-kD antigen in the immune deposits was followed by large electron-dense deposits, enhanced complement activity and proteinuria. The data suggest that in the growing immune complex lattice in the LRE. the 70-kD circulatory antigen by virtue of its small size, mobility and antigen cross-reactivity facilitates cross linking and coalescence of immune complexes, resulting in electron-dense immune deposits (EDD) formation which initiates complement activation and consequent proteinuria.
Changes in phenotypically distinct mucosal macrophage populations may be a prerequisite for the development of inflammatory bowel disease
Authors: ALLISON M C , POULTER L W
Published: July 2010

Previous studies have demonslraled the presence of much more marked macrophage heterogeneity in colonic mucosa affected by the idiopathic inflammatory bowel diseases (ulcerative colitis and Crohn’s disease) than in normal mucosa. This study examines the morphology, distribution and phenotypic expression of mucosal macrophage-like cells in biopsies from patients with idiopathic inflammatory bowel disease in comparison with disease control samples from patients with colonic infection or ischaemia. Approximately 80% of macrophage-like cells in histologically normal mucosa co-express the antigens recognized by the monoclonal antibodies RFDI (an interdigitating cell marker) and RFD7 (a marker for mature tissue macrophages). In idiopathic inflammatory bowel disease, the normal colonic macrophage population is partly replaced by cells staining positively with RFD7 alone, and, to a lesser extent, with RFDI+ dendritic cells. Sections from patients with infections and ischaemia exhibited epithelial HLA-DR positivity and infiltration of the lamina propria by a more heterogeneous population of macrophages than that seen in histologically normal mucosa. However, the displacement of the normal colonic macrophage phenotype by RFD7+ tissue macrophages occurred to a significantly greater extent in idiopathic inflammatory bowel disease than in disease control mucosa. A pathognomonic feature of the ulcerative colitis and Crohn’s colitis sections was the clustering of RFD9+ epithelioid cells at the bases of disrupted crypts and adjacent to areas of mucosal damage. It is concluded that a degree of macrophage heterogeneity and macrophage infiltration can occur as a non-specific response to colonic mucosal damage. The distinctive feature of idiopathic inflammatory bowel disease mucosa is the almost complete replacement of the normal colonic mucosal macrophage population by tissue macrophages and epithelioid cells, and this phenomenon may be important in promoting the development of a chronic inflammatory state
Analysis of the molecular mimicry between HLA-B27 and a bacterialOmpAprotein using synthetic peptides
Authors: YU D T Y , HAMACHI T , HAMACHI M , TRIBBICK G
Published: July 2010

In spite of a lack of sequence ‘homology’ between HLA-B27 and the bacterial OmpA outer membrane proteins, they both react with the Ye-2 monoclonal anti-HLA-B27 antibody. The Ye-2 antibody also reacted positively in ELISA with a synthetic peptide derived from the segment spanning residues 63–84 of B*2705. The critical peptide residues were determined by testing first with overlapping peptides, followed by a replacement set made according to the determined epitope. The results were compared with those with overlapping eight mers made to span a carboxyl fragment of the Escherichia coli OmpA. protein. They indicate the reason why Ye-2 reacts with both sets of peptides is because it has a preference for polymers of arginine.
Murine and human B cell epitope mapping of the Mycobacterium tuberculosis 10-kD heat shock protein using overlapping peptides
Authors: VERBON A , HARTSKEERL R A , KOLK A H J
Published: July 2010

The human immune response to the 10-kD M. tuberculosis protein was studied by a competition ELISA using monoclonal antibody (MoAb)SA-12. Twenly-five per cent of the sera from 20 patients with tuberculosis and none from 21 control subjects inhibited binding of SA-12 to the 10-kD antigen. To characterize theantigenic parts of the 10-kD antigen, overlapping decapeptides according to the amino acid sequence of the 10-kD protein were synthesized. In total. 91 sequential decapeptides, with an overlap of nine amino acids, were tested in ELISA with MoAb SA-12, human and murine sera (PEP scan). SA-12 recognized the amino acid sequence WDEDGEK (amino acid 50 56). All human sera, from patients with tuberculosis as well as from control subjects, gave almost identical undulating patterns of reactivity with the decapeptides. No relationship was found between the ability of the patients’ sera to inhibit binding of MoAb SA-12 and the binding of these sera to the decapeptides comprising the epitope recognized by SA-12 in the PEP scan. Apparently, antibodies in patients’ sera against the 10-kD protein are predominantly directed against discontinuous epitopes and, consequently, the continuous epitopes as presented in the PEP scan are not suitable to discriminate between patients with tuberculosis and control subjects. In the PEP scan, sera from BALB/c mice, both non-immunized and immunized with either live M, tuberculosis or the 10-kD protein gave similar patterns of reactivity, albeit different from the patterns obtained with the human sera. However, after immunization of the mice, clearly increased levels of antibodies to primary structures of the 10-kD protein were observed.
Lymphocyte subpopulations in pyogranulomas of caseous lymphadenitis
Authors: WALKER J , JACKSON H , BRANDON M R , MEEUSEN E
Published: July 2010

Pyogranulomas of ovine caseous lymphadenitis (CLA) are encapsulated lesions resulting from infections with Corynebacterium pseudotuberculosis, a bacterial pathogen able to grow within macrophages. Immunohistology of CLA lesions showed a band of lymphocytes lining the inside of the collagen capsule in intimate contact with necrotic tissue, the intracapsular lymphocytes being organized into three layers. The innermost layer, immediately adjacent to the central necrotic tissue consisted of a narrow band of MHC class II staining macrophages. Cells staining for CD4. CD8 and γδ T cell markers were unevenly distributed throughout the lymphoid layer, tending to be more numerous immediately external to the macrophage layer. The intracapsular lymphoid tissue contained a high proportion of CD8+ lymphocytes (CD4:CD8, 1.5:1) and of γδ lymphocytes (CD4:CD8:γδ, 1:0.7:0.8). External to the T cell-rich zone and adjacent to the surrounding collagen capsule was a dense band of cells, a proportion of which stained atypically for CD45R and were tentatively identified as B cells. CDS+ and γδ+ T cells showed similar distributions and their relative abundance, compared with CD4+ T cells, was a distinguishing feature of the CLA lesion. Staining for factor VIII-related antigen clearly showed endothelial venules throughout the intracapsular lymphoid tissue. The presence of endolhelial vcnules and the organized architecture of the lymphoid tissue teleologically argues that lymphocytes are continually recruited into chronic CLA lesions and play an important role in the ongoing disease process.
Human cerebral malaria: characterization of malarial antibodies in cerebrospinal fluid
Authors: MITRA S , RAVINDRAN B , DAS B K , DAS R K , DAS P K , RATH R N
Published: July 2010

Anti-malarial antibodies were quantified in cerebrospinal fluid (CSF) of 17 cases of cerebral malaria, 16 presumptive cases (no demonstrable parasitaemia in peripheral blood but responding to i.v. quinine therapy) of cerebral malaria, and 15 controls. A schizont-enriched Plasmodium knowlesi antigen was used in an ELISA. Anti-malarial antibodies of IgA and IgM isotypes were not detectable in most of the CSF samples analysed, although serum antibody titres were high. However, 88% of CSF from cerebral malaria and 56% of presumptive cerebral malaria cases had significant levels of IgG anti-malarial antibodies in comparison to control CSF. The antibody levels did not correlate with the severity of coma but correlated well with the duration of coma. The CSF malarial antibody titres were independent of degree of parasitaemia. The possible role of CSF anti-malarial antibodies in cerebral malaria in the light of recent demonstrations of intrathecal synthesis of immunoglobulins and deposition of immune complex in cerebral tissues is discussed.
Cytotoxicity of human natural killer (NK) cell subsets forPlasmodium falciparumerythrocytic schizonts: stimulation by cytokines and inhibition by neomycin
Authors: ORAGO A S S , FACER C A
Published: July 2010

Quantification of human peripheral blood NK subsets has been made in a group of Kenyan adults and children with acute P. falciparum malaria. Results were compared with data obtained from three age- and sex-matched control cohorts: parasitaemic but asymptomatic children: aparasitaemic children and adults; and adult Caucasians with no previous history of malaria. Separated NK subsets were tested in vitro for cytotoxicity to erythrocytic sehizonts of P. falciparum in the presence and absence of cytokines. There was a statistically significant quantitative and qualitative depression of the CD3- CD56+ subset in patients with acute malaria and this was accompanied by an expansion of the ‘non-functional’ CD3- CD57+ CD16- CD56- subset. Both CD3- CD16+ and CD3- CD56+ NK cells from all patients and donors lysed schizonts. and this cytotoxicity was enhanced by the addition of recombinant interferon-alpha and or IL-2, notably with the CD3- CD56+ subset. Interestingly, asymptomatic donors had the highest levels of CD3- CD56+ NK cells, which also demonstrated an enhanced response to cytokine stimulation. Cytotoxicity to schizonts was accompanied by the release of soluble NK cell lytic factors. Neomycin suppressed cytotoxicity in a dose-dependent manner, Indicating that the lysis of schizonts by NK cells involves phospholipase C-mcdiated phosphoinositide metabolism. Our findings define a role for NK cells in immunity to malaria through the lysis of infected erythrocytcs as a first-line defence against the parasite.
Mapping of epitopes on Ul snRNP polypeptide A with synthetic peptides and autoimmune sera
Authors: BARAKAT S , BRIAND J P , ABUAF N , VAN REGENMORTEL M H V , MULLER S
Published: July 2010

The ability of synthetic peptides encompassing almost the entire sequence of snRNP U1A polypeptide to be recognized in ELISA by sera of autoimmune patients was investigated. Sera from 18 patients with mixed connective tissue disease (MCTD), 145 with systemic lupus erythematosus (SLE) and 120 with other rheumatic autoimmune diseases were tested with 13 overlapping peptides. Among them, peptide 257–282 and, to a lower extent, peptide 1–11 were recognized by MCTD, SLE and Sjogren’s syndrome sera. In contrast, peptide 35–58 was recognized by 94% of MCTD and only 19% of SLE sera. It did not react with any of the other patient sera. The ELISA results were compared with the pattern of reactivity observed in immunoblotting. The results indicate that peptide 35–58 probably contains a major epitope recognized by MCTD autoantibodies. It is noteworthy that in snRNP particles, this region of U1A interacts with RNA and presents only limited homology with the corresponding sequence 32–50 of U2B”.
Detection and occurrence of the 60- and 52-kD Ro (SS-A) antigens and of autoantibodies against these proteins
Authors: SLOBBE R L , PRUIJN G J M , DAMEN W G M , VAN DER KEMP J W C M , VAN VENROOIJ W J
Published: July 2010

The simultaneous detection of anti-La. anti-60-kD Ro and anti-52-kD Ro antibodies by immunoblotting is greatly improved by changing the crosslinking level in the gel to an acrylamide/bisacrylamide ratio of 19:1. Using this method for the analysis of a number of systemic lupus erythematosus (SLE) and Sjögren’s syndrome patient sera it was observed that antibody to the 52-kD Ro protein without anti-60-kD Ro antibody was restricted to Sjogren’s syndrome patients (9/26), whereas antibody to the 60-kD Ro protein without contaminating anti-52-kD Ro antibody was only found in SLE patients (8/38). Moreover, in Sjogren’s sydrome patient sera anti-Ro antibody was found only in combination with anti-La antibody (20/26). whereas in SLE patient sera anti-Ro antibody could be found without detectable anti-La specificity (4/38). Double immunofluorescence microscopy revealed that the 52-kD Ro and the 60-kD Ro proteins co-localize in the cytoplasm as well as in the nucleus, whereas immunoprecipitation of [32P]-labelled HeLa cell extract with monospecific anti-52-kD Ro and anti-60-kD Ro sera showed that both proteins are associated with the Ro RNAs. These data suggest the presence of both the 52-kD and the 60-kD Ro proteins in the same ribonucleoprolein complexes. To study the evolutionary conservation of the 52-kD Ro, the 60-kD Ro and the La proteins, extracts of cell lines derived from various mammalian species were analysed on Western blols using monospecific human antibodies. In contrast to the 60-kD Ro and the La antigens which are well conserved in evolution, the 52-kD Ro antigen could be detected in primate cells only by this immunological approach.
New monoclonal anti-human Fc gamma receptor II antibodies induce platelet aggregation
Authors: NOMURA S , YAMAGUCHI K , KIDO H , KAWAKATSU T , IWATA K , FUKUROI T , SUZUKI M , YANABU M , SOGA T , NAGATA H , KOKAWA T , YASUNAGA K
Published: July 2010

We developed two new monoclonal antibodies, designaled NNKY3-2 and NNKY4-7, that recognized a 40-kD platelet protein. They appeared to be monoclonal anti-Fc gamma receptor II (FC-/RII) antibodies from the results of flow cytometric binding inhibition studies using another monoclonal anti-FcγRII antibody (2E1). The addition of NNKY3-2 or NNKY4-7 to platetet-rich plasma (PRP) led to a typical aggregation pattern preceded by a lag phase, but their addition to washed platelets did not induce aggregation. The aggregation of PRP by these antibodies was inhibited by prostaglandin E1 (PGE1) or staurosporine (protein kinase C inhibitor), whereas it was only slightly affected by a monoclonal anti-GPIIb/IIIa antibody or Arg-Gly-Asp-Ser, Furthermore, these antibodies induced the aggregation of washed platelets plus normal serum, but not that of washed platelets plus heat-treatcd serum (destruction of complement activity). These results suggest that NNKY3-2 or NNKY4-7-induced aggregation involves an unusual pathway independent of fibrinogen, and that the important factor is the participation of complement. NNKY3-2 and NNKY4-7 may be useful to study the relationship between autoantibodies, the Fc receptor, and complement in idiopathic thrombocytopenic purpura.
FcγRIIα: sequencing of the ligand binding domain in systemic lupus erythematosus patients
Authors: JAZWINSKA E C , BANYER J L , GATENBY P A , SERJEANTSON S W
Published: July 2010

The receptor for the Fc portion of IgG (FcγR) is involved in the slow clearance of immune complexes. To perform this role it must be cross-linked by IgG bound in its extracellular domains. It has been suggested that defective FcγR-mediated clearance of immune complexes may play a role in the pathogenesis of autoimmune connective tissue disorders. We have sequenced DNA encoding the second extracellular domain of FcγRIIα in six patients with SLE, to investigate whether point mutations may be responsible for encoding a defect in the IgG binding capacity of the receptor. We were able to identify the point mutation which discriminates high and low responder genotypes but found no other change from the published DNA sequence for this region.
Acquired resistance to type II collagen-induced arthritis in rhesus monkeys is reflected by a T cell low-responsiveness to the antigen
Authors: BAKKER N P M , VAN ERCK M G M , HART L A T , JONKER M
Published: July 2010

Ten out of 14 rhesus monkeys developed arthritis after a single immunization with bovine type II collagen (B-CII). In contrast to primary resistant monkeys, arthritic animals showed a B-CII specific T cell proliferation during the induction phase of the disease. All surviving animals showed a full remission of the disease. Two monkeys acquired resistance to collagen-induced arthritis (CIA) after one period of disease, but in three animals a booster immunization with B-CII induced a slight flare-up. It is demonstrated that B-CII immunized rhesus monkeys have the capacity to restore resistance to CIA. The development of resistance to CIA is reflected by a decreased T cell responsiveness to B-CII, It is shown that the lack of IL-2 plays a role in B-CII-induced T cell low-responsiveness. A potential role of CD8+ T cells in the down regulation of the T cell response to B-CII is discussed.
The effect of FK506 and cyclosporin A on antigen-induced arthritis
Authors: BLACKHAM A , GRIFFITHS R J
Published: July 2010

FK506 and cyclosporin A inhibited the development of antigen-induced arthritis in the rat and rabbit. FK506 was five times more potent than cyclosporin A in the rat and approximately 20 times more potent in the rabbit. FK506 was effective in both species if administered either from the day of intra-articular administration or antigen or when the arthritis was established. In the rabbit, arthritis returned when administration of FK506 was stopped. FK506 (10 mg/ kg day) caused renal damage which was not observed at a dose of 2.5 mg/kg/day. Both of these doses were equally effective at inhibiting the arthritis. The conclusion from these studies is that FK506 is a more effective antiarthritic agent than cyclosporin A and that a pronounced therapeutic effect can be achieved at non-toxic doses of the drug.
Cell-bound IgE and increased expression of Faɛ-receptors on dendritic cells in cutaneous infiltrates of mycosis fungoides
Authors: PREESMAN A H , VAN DE WINKEL J G J , MAGNUSSON C G M , TOONSTRA J , VAN DER PUTTE S C J , VAN VLOTEN W A
Published: July 2010

Skin biopsies of 31 non-atopic patients, 20 with mycosis fungoides, six with psoriasis and five with contact dermatitis, and of five non-atopic healthy controls were compared for the presence of cell-bound IgE and vacant IgE binding sites. IgE+ cells were demonstrated in the cutaneous infiltrate of nine (45%) patients with mycosis fungoides, two (33%) with psoriasis and one (20%) with contact dermatitis. Following pre-incubation of skin sections wilh IgE myeloma protein to saturate vacant IgE-binding sites, 14 out of 16 patients (88%) with stage I mycosis fungoides, five (83%) patients with psoriasis and one (20%) with contact dermatitis showed an increase in the number of IgE+ cells. While cell-bound IgE was positively related to serum IgE levels the expression of IgE-binding sites was not. All IgE+ cells were HLA-DR+ dendritic cells identified as either macrophages (CD68+, CDI4+) or Langerhans cells (CDl+). Skin biopsies of non-atopic healthy controls or clinically uninvolved skin in mycosis fungoides had neither any IgE+ cells nor any vacant binding sites. Inhibition studies with IgGl, IgG4 and IgE myeloma proteins as well as with several enzymatic fragments of igE demonstrated that IgE interacled with Fcɛ-receptors through isotype-specific structures on the Fcɛ-fragment. Four anti-CD23 monoclonal antibodies, however, were unuble to stain vacant Fcɛ-receptors nor could they block IgE-binding. We hypothesize that locally-secreted lymphokines, like IL-4 or interferon-γ, induce Fcɛ-receptors on dendritic cells in the cutaneous infiltrate and that these receptors become occupied in parallel wilh elevated serum IgE levels.