bluuebunny/crossref_metadata_2025 huggingface dataset:
Polymorphic MHC ancestral haplotypes affect the activity of tumour necrosis factor-alpha
Authors: ABRAHAM L J , FRENCH M A H , DAWKINS R L
Published: July 2010

It remains unclear which MHC loci are involved in susceptibility to autoimmune diseases and immune deficiencies. We have chosen to evaluate whether different alleles of tumour necrosis factor-alpha (TNF-α) are important, as TNF has been implicated in the etiology of many immunological disorders. We have shown previously that a restriction fragment length polymorphism in the TNF region correlates with MHC ancestral haplotypes. We therefore examined the effect of ancestral haplotype on the activity of TNF-α in culture supernatants of lymphoblastoid cell lines. The results demonstrate that TNF-α activity in supernatants of 8.1 (A1, B8, DR3) cell lines was higher than that present in the supernatants from cells homozygous for eight different MHC ancestral haplotypes, and indicate that polymorphisms in TNF-α, or in other MHC genes that regulate TNF, may be responsible for the 8.1 phenotype.
Increased spontaneous secretion of rheumatoid factor by intestinal lamina propria mononuclear cells from Crohn's disease but not ulcerative colitis patients
Authors: MacDERMOTT R P , SCHREIBER S , NASH G S , KOOPMAN W J
Published: July 2010

Increased levels of rheumatoid factors (RF) have been observed in the scrum of Crohn's disease but not ulcerative colitis patients, and have been proposed to relate to an increased state of intestinal lymphocyte activation. We have therefore examined the spontaneous in vitro secretion of RF by intestinal lamina propria mononuclear cells (MNC) isolated from specimens from control and inflammatory bowel disease (Crohn's disease, ulcerative colitis) patients. Normal intestinal lamina propria MNC spontaneously secrete rheumatoid factors of different isotypes during 14 days of in vitro culture (9·7 ng/ml IgA RF, 11·6 ng/ml IgM RF and 64·6 ng/ml IgA anti-Fc (IgG)). In matched studies intestinal MNC isolated from normal large bowel exhibited significantly greater levels of RF synthesis and secretion in vitro than normal small bowel intestinal MNC. A large increase in spontaneous RF secretion was observed from Crohn's disease intestinal MNC (21·4 ng/ml IgA RF, 21·4 ng/ml IgM RF, and 108·15 ng/ml IgA anti-Fc (IgG)), when compared with normal controls. The amount of RF secreted was dependent on the amount of inflammatory activity of the bowel specimens, from which the MNC were isolated (198·3 ng/ml of IgA anti-Fc(IgG) from involved versus 50·0 ng/ml from matched non-involved tissue). Ulcerative colitis MNC released decreased amounts of RF(7I ng/ml IgA RF, 6·2 ng/ml IgM RF, and 42·3 ng/ml IgA anti-Fc (IgG)). These observations using isolated intestinal MNC may explain the findings of RF changes in the sera of inflammatory bowel disease patients. Our observations support the hypothesis of a heightened stale of activation in normal intestinal lamina propria MNC, which is further increased in active Crohn's disease. The dissimilarities observed between Crohn's disease and ulcerative colitis may indicate fundamental differences in disease pathophysiology and will lead to further studies exploring intestinal immunoregulatory properties of RF.
The effects of cytokines on metalloproteinase inhibitors (TIMP) and collagenase production by human chondrocytes and TIMP production by synovial cells and endothelial cells
Authors: SHINGU M , NAGAI Y , ISAYAMA T , NAONO T , NOBUNAGA M , NAGAI Y
Published: July 2010

It has been suggested that IL-1 produces cartilage matrix degradation by metalloproteinases such as collagenase, and that such degradation is regulated by metalloproteinase inhibitors (TIMP). Therefore, the balance between collagenase and TIMP is an important factor for tissue destruction in inflammatory joints. In the present study the effects of cytokines on collagenase and TIMP production in chondrocytes as welt as the effects of cytokines on TIMP production in connective tissue cells were studied. IL-Iβ inhibited TIMP production in endothelial cells while enhancing TIMP production in synovial cells and chondrocytes. In addition, tumour necrosis factor-alpha (TNF-α) significantly inhibited and IL-6 significantly enhanced TIMP production in endothelial cells, synovial cells and chondrocytes. In the chondrocyte supernatant, collagenase activity/TIMP ratio was significantly elevated by the addition of either IL-lβ or TNF-α to the cells, whereas the ratio was significantly decreased by IL-6. These results suggest that the cytokine effects on TIMP production are different among the different cell types, and that either IL-lβ or TNF-α induce cartilage matrix degradation by disrupting the collagenase/TIMP balance, while, on the other hand, IL-6 protects the tissue through an opposite effect.
Enhand secretion of tumour necrosis factor-alpha, IL-6, and IL-1β by isolated lamina ropria monouclear cells from patients with ulcretive cilitis and Crohn's disease
Authors: REINECKER H-C , STEFFEN M , WITTHOEFT T , PFLUEGER I , SCHREIBER S , MacDERMOTT R P , RAEDLER A
Published: July 2010

The perpetuation of inflammation in ulcerative colitis and Crohn's disease may be regulated in part by an increased secretion of proinflammatory cytokines due to either an appropriate response to initial stimulating agents, and/or due to an impaired down-regulalion of cylokine secretion. The aim of this study was to determine the secretion patterns of the proinflammatory cytokines tumour necrosis factor-alpha (TNF-α), IL-6 and IL-1β, from isolated lamina propria mononuclear cells (LPMNC) isolated from colonic biopsies from patients with untreated ulcerative colitis or Crohn's disease. LPMNC isoiated from involved inflammatory bowel disease (lBD) mucosa spontaneously produced increased amounts of TNF-α, IL-6, and IL-1β. The TNF-α secretion from IBD LPMNC could be further enhanced by pokeweed mitogen stimulation. The secretion patterns of TNF-α and IL-1β by LPMNC from patients with either uicerative colitis or Crohn's disease demonstrated a close correlation with the degree of tissue involvement and mucosal inflammation. LPMNC from noninvolved ulcerative colitis mucosa secreted markedly increased leveis of IL-6 compared with noninvolved Crohn's disease mucosa or control mucosa. The heightened lL-6 secretion from LPMNC from non-involved ulcerative colitis mucosa without visible or microscopic signs of inflammation indicates that the pathophysiologie mechanisms involved in the initiation of inflammation may differ between ulceralive colitis and Crohn's disease. The determination of proinflammatory cytokine secretion by isolated LPMNC from colonoseopic biopsies may be a sensitive method for monitoring the severity of mucosal inflammation in IBD patients.
B cell activation in clinically quiescent systemic lupus erythematosus (SLE) is related to immunoglobulin levels, but not to levels of anti-dsDNA, nor to concurrent T cell activation
Authors: SPRONK P E , v.d. GUN B T F , LIMBURG P C , KALLENBERG C G M
Published: July 2010

In clinically quiescent SLE hypergammaglobulinaemia, presence of autoantibodies, and increased soluble IL-2 receptors (sIL-2R) have been reported, suggesting persistent B as well as T cell activation. In contrast, the primary immune response lo test antigens is markedly decreased. To analyse these phenomena at a cellular level, we undertook a cross-sectional study on 13 non-active SLE patients and 15 controls. We determined the composition of lymphocyte subsets with special attention to activation markers (CD25, HLA-DR, CD38) and the presence of naive T cells (CD45RO-), and related those findings to serological parameters. In non-active SLE patients the expression of activation markers on B cells and T cells was higher than in normal controls (P≤ 0·02), but was not interrelated. Percentages of activated B cells in SLB were related lo levels of total IgG (P < 0·02)and IgM (P < 0·02) but not to anti-dsDNA, suggesting a disordered immune system also in clinically quiescent SLE. Numbers of CD4+ cells (P < 0·001) and CD4+CD45RO- cells (P < 0·05) were decreased. The latter finding might explain the anergy to primary test antigens in clinically quiescent SLE.
Immunological responses of Gambians in relation to clinical stage of HIV-2 disease
Authors: WHITTLE H , EGBOGA A , TODD J , MORGAN G , ROLFE M , SABALLY S , WILKINS A , CORRAH T
Published: July 2010

This study describes a broad spectrum of cellular and antibody-mediated immune responses found in 28 asymptomatic and 37 symptomatic Gambian patients with HIV-2 infection. It shows that these responses vary according to the stage of infection as described by three clinical staging systems. The first system was a local one based on the signs used for the WHO Bangui clinical definition of AIDS, the second, suggested by WHO, was based on a performance scale, and the third was that used by the Centre for Disease Control. Asymptomatic patients had significantly lower mean CD4 counts, lymphoproliferative and interferon-gamma (IFN-γ) responses and lower IgG and IgM antibody responses to keyhole limpet haemocyanin (KLH) than controls. These measurements and the size of the skin test reaction to purified protein derivative (PPD) or Candida antigen declined significantly according to the stage of infection. Mean values of the serological markers β2-microglobulin and neopterin and antibody titres to Epstein-Barr virus capsid antigen (EBVCA) rose significantly according to severity of disease. The Gambian or WHO clinical staging systems, which are easy and cheap to apply, may serve as an alternative to sophisticated and expensive immunological measurements when trying to stage disease and predict prognosis.
Antibodies to proteinase 3 increase adhesion of neutrophils to human endothelial cells
Authors: MAYET W-J , MEYER ZUM BÜSCHENFELDE K-H
Published: July 2010

The detection of anti-neutrophil cytoplasmic antibodies (ANCA), especially those with specificity for proteinase 3, is important in the diagnosis and in monitoring disease activity of Wegener's granulomatosis and related vasculitides. An ubiquitous feature of all ANCA-associated acute vascular injury is lytic necrosis. Adhesion of neutrophils to endothelium is a fundamental early step of the inflammatory response. Recently we were able to show that ANCA recognize their target antigen (proteinase 3) translocated into the membrane of human endothelial cells. The aim of this study was to investigate the effect of ANCA on the adhesion of neutrophils to human endothelial cells. Incubation of endothelial cells with affinity-purified antibodies to proteinase 3 (IgG- and F(ab′)2-fractions) led to a marked increase of neutrophil adhesion, with a peak after 4 h and a rapid decrease after 8 h. This effect could be inhibited by preincubation of the endothelial cells with an antibody to endothelial leucocyte adhesion molecule-1 (ELAM-1). Incubation with antibodies to proteinase 3 also led to an incrcase of endothelial ELAM-1 expression as measured in a cyto-ELISA and by flow cytometry. Our data demonstrate a direct effect of ANCA on neutrophil endothelial interactions. The enhanced adhesion of neutrophils occurs time-dependently via induction of ELAM-1 expression on the surface of endothelial cells. Our data give a hint of an ANCA-mediated mechanism of endothelial injury via induction of neutrophil adhesion to vascular endothelium in Wegener's granulomatosis and other ANCA-related vasculitides.
Glycation increases the vascular clearance rate of IgG in mice
Authors: KENNEDY D M , SKILLEN A W , SELF C H
Published: July 2010

As elevated levels of glycated IgG have been detected in the plasma of diabetics we have investigated whether glycation of IgG affects its vascular clearance rate, using a mouse model system. Polyclonal mouse IgG was aseptically incubated for 14–19 days with 0.5 m glucose in 0.1 m phosphate buffer (pH7.4) at 37 C, As control, IgG was incubated under identical conditions but with no added glucose. After incubation, both forms were labelled with 125I and injected intravenously into BALB/c mice. The rate of vascular clearance of the glycated IgG was found to be significantly higher than the control IgG in the periods 524 h (P<0.001,n= 6) and 24–48 h (P<0.01, n=6) after injection. After 2–3 days the mice were killed and the major organs were harvested. With glycated IgG there was a significant increase in the 125I accumulated in the kidney (P<0.02). In later experiments, dual labelling with 131I and 1251 allowed mixtures of glycated and unglycated IgG to be injected into the same mouse so that the vascular clearance of both forms of IgG could be followed simultaneously. These experiments confirmed that glycation of the IgG significantly increases its vascular clearance rate.
Identification of human T cell epitopes in theMycobacterium lepraeheat shock protein 70-kD antigen
Authors: ADAMS E , BRITTON W J , MORGAN A , GOODSALL A L , BASTEN A
Published: July 2010

In a number of pathogens, heat shock proteins (hsp) stimulate humoral and cellular immune responses despite significant sequence identity with host hsp. The 70-kD hsp of Mycobacterium leprae, whieh shares 47% identity with human hsp70 at the protein level, elicited a T cell response in most Myco. bovis (bacille Calmette Guérin (BCG)) vaccinees as well as leprosy and tuberculosis patients and their contacts. In order to locate T cell epitopes, DNA fragments encoding portions of the 70-kD hsp were expressed in the vector pGEX-2T and tested for T cell reactivity in an in vitro proliferative assay. Cultures of peripheral blood mononuclear cells (PBMC) from BCG vaccinees indicated that the C-terminal half of the molecule contained multiple T cell epitopes, as the T cells from a majority of Myco. leprae hsp70-reactive individuals responded to C-344. Lower proportions of patients with paucibacillary leprosy (36%) and tuberculosis patients (16%) responded to C-344. The smaller C-142 fragment which includes the terminal 70 residues unique to Myco. leprae and is the target for the human antibody response elicited a cellular response in few patients and no vaccinees. In order to map T cell epitopes, two series of synthetic peptides encompassing the region 278–502 were prepared. Using overlapping 12mer and 20mer peptides, this region of the molecule was found to contain several potential T cell epitopes. The longer peptides gave a clearer indication of reactive sequences including regions of the molecule which were not identified with the 12mer peptides. Fine mapping of reactive peptide pools using the 12mer peptides identified two T cell epitopes. Although both were located in regions of the molecule shared with Myco. tuberculosis, one appeared to be cross-reactive with the equivalent human sequence, and thus has the potential to initiate autoimmune responses.
Anti-CD5 therapy decreases severity of established disease in collagen type II-induced arthritis in DBA/1 mice
Authors: PLATER-ZYBERK C , TAYLOR P C , BLAYLOCK M G , MAINI R N
Published: July 2010

Collagen-induced arthritis has been widely used as an animal model of rheumatoid arthritis. We have used this model with a view to determining potential therapeutic targets for the treatment of human disease. To do this we have attempted to modulate the progression of established arthritis over a 10-day time period following the first appearance of disease, by i.p. injection of one of three different MoAbs. These consist of a rat IgG2a specific for the CD5 antigen expressed on all T cells and a subpopulation of B cells, a mouse IgG2b recognizing the CD72 antigen, and a rat IgM specific for the B220 molecule, CD72 and B220 both being expressed on all B cells. None of the three MoAbs had depleting activity in vivo. The progression of arthritis was monitored both clinically, and histologically. The effects of treatment with anti-CD5 and anti-CD72 antibodies were compared with control antibodies of the same species class and subclass. In the case of anti-B220 antibodies, the effects of treatment were compared with administration of FBS. Of these MoAbs, only treatment with anti-CD5 resulted in disease amelioration with significant decrease in disease severity in 60% of the animals. These changes became apparent 6 days after initiation of treatment. There were no significant differences in serum levels of IgG antibodies to native bovine collagen type II between the groups of treated and control mice. Possible mechanisms underlying the modification of disease expression following treatment with anti-CD5 MoAb are discussed.
Perforin expression by thyroid-infiltrating T cells in autoimmune thyroid disease
Authors: WU Z , PODACK E R , McKENZIE J M , OLSEN K J , ZAKARIJA M
Published: July 2010

Infiltration of the thyroid gland by lymphocytes is a hall-mark of autoimmune thyroid disease; it is particularly evident in Hashimoto's thyroiditis but is also seen in most patients with Graves’ disease. Infiltrating cells are comprised primarily of T lymphocytes., of which only a minority appears to be activated. Their precise pathogenic role is largely unknown. Since perforin has been a marker for functionally activated cytotoxic T cells in situ we elected to assess the presence of perforin-containing cells in thyroid-infiltrating lymphocytes and establish their phenotype. Cells were isolated from seven subtotal thyroidectomy specimens, five from patients with Graves” disease and two with Hashimoto's thyroiditis. The novel findings were as follows: CD4+ perforin-containing T cells occurred only in Hashimoto's glands, suggesting a class II-restricted component of cytotoxicity; in Graves' disease, and to a lesser extent in Hashimoto's, perforin-expressing cells were primarily T cell receptor αβ+ CD4- CD8- (double negative); double negative perforin-containing cells in peripheral blood of normal individuals were largely γδ+T cells. In Hashimoto's samples, the predominant population of T cells expressing perforin was CD8+. By comparison, in studies of the synovial fluid of knee joints from patients with rheumatoid arthritis only a minor population of the perforin-containing cells was double-negative. The data suggest significant differences in cytotoxic autoimmune mechanisms between the two autoimmune thyroid diseases. Functional characterization of double-negative T cells is necessary to define their role in autoimmunity.
Characterization of neural cell adhesion molecule (NCAM) expression in thyroid follicular cells: Induction by cytokines and over expression in autoimmune glands
Authors: VARGAS F , TOLOSA F , SOSPEDRA M , CÁTALFAMO M , LUCAS-MARTÍN A , OBIOLS G , PUJOL-BORRELL R
Published: July 2010

NCAM (CD56) is a cell surface glycoprotein of the immunoglobulin superfamily expressed on neuroendocrine and natural killer (NK) cells which has considerable molecular heterogeneity due to differential splicing and post-translational modifications. NCAM has been detected in the thyroid follicular cells (thyrocytes) immunohistological. We report here the molecular form, the modulation by cytokines and the levels of expression in thyroid pathology. By using a panel of MoAbs to NCAM on Western blots from thyrocyte extract we have determined that these cells express the 140- and 180-kD forms of NCAM, Exposure of primary cultures of thyrocytes to interferon-gamma (IFN-Γ), and even more, to the combination of IFN-Γ plus tumour necrosis factor-alpha (TNF-α) induced a clear increase in the expression of NCAM as assessed by FACS analysis. NCAM expression in thyrocytes was assessed by immunofluorescence in 59 surgical specimens of thyroid glands, and was found increased in 11/17 (64%) of Graves’, in 5/25 (20%) of multinodular goitre (MNG) and in occasional adenoma glands. No correlation was found with the expression of HLA class I. class II or the degree of lymphocytic infiltration scored in adjacent sections, but it was often seen in areas infiltrated by macrophages. In conclusion, NCAM is an adhesion molecule whose expression is clearly increased in thyrocytes in autoimmune glands, probably as a consequence of exposure to cytokines locally released. Since one of the forms of NCAM expressed by thyrocytes has the capability to generate intracellular signal it may play a role in normal thyroid function. In addition, NCAM may facilitate the recognition of thyrocytes by lymphocytes, particularly by NK CD56+ lymphocytes.
Failure in IgA secretion by surface IgA-positive B cells in common variable immunodeficiency (CVID)
Authors: SCOTT L J , BRYANT A , WEBSTER A D B , FARRANT J
Published: July 2010

CVID is an acquired disease with low serum immunoglobulins. Patients are classified into three groups on the ability of their B cells to secrete IgM and IgG in vitro in response to IL-2. We have now extended this patient classification to include IgA secretion and to assess whether the patients have IgA+ B cells in the circulation. B cells from almost all of the CVID patients studied were unable to secrete any IgA in vitro, with or without IL-2, although all B cells tested from normal donors showed some spontaneous IgA secretion. Thus, the defect with IgA secretion is more profound than with the secretion of IgM or IgG. Despite this failure to secrete IgA, using flow cytometry techniques all CVID patients tested were found to have IgA+ cells (expressed as a percentage of B cells) within the normal range. This suggests that CVID may involve a defect in the secretion of immunoglobulin isotypes rather than a defect in isotype switching.
Antineutrophil cytoplasm antibodies (ANCA) of IgA isotype in adult Henoch-Schönlein purpura
Authors: RONDA N , ESNAULT V L M , LAYWARD L , SEPE V , ALLEN A , FEHHALLY J , LOCKWOOD C M
Published: July 2010

ANCA are associated with certain forms of systemic vasculitis, and have been reported previously to be of the IgG and IgM isotype. We examined ihc possible association between IgA ANCA and the IgA-related diseases Henoch-Schönlein purpura (HSP) and IgA nephropathy (IgAN). IgA and IgG ANCA were detected by isotype-specific solid-phase assays with a crude neutrophil extract, and their presence was confirmed by antigen-specific fluid-phase competitive inhibition tests and by indirect immunofluorescence. The possible interference by IgA rheumatoid factor was excluded. IgA ANCA were detected in sera from 11/I4 HSP patients (79%). from 1/30 IgAN patients (3%), from 1/40 patients with vasculitides classically associated with IgG ANCA (2.5%), and in none or 60 sera from healthy blood donors. IgG ANCA were present with IgA ANCA in three patients with HSP. Only one HSP serum had anti-myeloperoxidase (MPO) activity by both IgA and IgG isotype-specific ELISA, and none was positive for proteinase 3 (PR3). Western blot analysis performed with neutrophil extract showed that the four strongest IgA ANCA-positive HSP sera reacted with a 51 -kD protein; Western blot performed on cellular fractions showed that this protein is primarily membrane-associated, and different from fibronectin. Our study suggests that adult HSP is closely associated with circulating IgA ANCA. which may be directed against a different autoantigen than that recognized by IgG ANCA.
Production and secretion of interferon-gamma (IFN-γ) in children with atopic dermatitis
Authors: TANG M , KEMP A
Published: July 2010

IFN-γ is known to be a major inhibitor of IgE synthesis in vitro. Recent studies demonstrating reduced production of IFN-γ in children and adults with atopic dermatitis and elevated serum IgE suggest a similar role for this cytokine in vivo. The reasons for this reduced IFN-γ production are not known. One possibility is Ibal atopic individuals have a reduced population of cells producing IFN-γ in vivo. Using a fluorescence-labelled antibody to detect intracellular IFN-γ. the percentage of IFN-γ-producing cells was determined in children with atopic dermatitis and in non-atopic controls. Children with atopic dermatitis had a greater percentage of IFN-γ-producing ceils in unstimulated cultures compared with controls, indicating in vivo activation of lymphocytes in the atopic group. This could reflect the significant degree of inflammation present in these children, or the presence of bacterial infection or colonization. Although secretion of IFN-γ after stimulation with phorbol myristate acetate(PMA)/Ca was significantly lower in children with atopic dermatitis compared with controls, the percentage of IFN-γ-producing cells in the stimulated cultures from this group was equivalent to controls. This demonstrates that the reduced ability of atopic children to secrete IFN-γin vitro does not relate lo a lack of IFN-γ-producing cells, but to a difference in the regulation of IFN-γ production beyond the stage of signal transduction.
IgG glycosylation in autoimmune-prone strains of mice
Authors: BODMAN K B , HUTCHINGS P R , JEDDI P A , DELVES P J , ROOK G A W , SUMAR N , ROITT M , LYDYARD P M
Published: July 2010

The relationship between increased levels of IgG oligosaccharide chains lacking galactose (G0) and the development of rheumatoid arthritis is unclear. In order to further our understanding of the observed correlation between raised serum G0 and arthritis, we have studied G0 levels in arthritis prone and non-susceptible (i.e., non-arthritis-prone) mice and the effects on G0 of mycobacterial antigens, which have been postulated to play a role in the early events leading to the development of arthritis. We have shown that different age-matched mouse strains have characteristic “resting” levels of G0 which (in six out of seven strains of mice) increase with age. We have also shown that these increases can be enhanced by immunization of arthritis-prone strains of mice with an adjuvant containing mycobacteria (Freund's complete adjuvant (FCA)), suggesting that deflects in the ability to regulate these G0 changes may be related to susceptibility to arthritis.
The effect of anti-CD4 on helper function of CD4,45RA+ versus CD4,45RO+ T cells
Authors: WANG J , YAN T , SIMMER B , EMMRICH F
Published: July 2010

Here we have investigated and compared the effects oranti-CD4 on helper function of CD4, 45RA+ versus CD4, 45RO+ Tcells. Only CD4, 45RO+ cells, but not CD4, 45RA+ cells were able lo promote B cell differentiation resulting in immunoglobulin production in vitro(IgM as well as IgG) which could be inhibited by anti-CD4 MoAbs (MAX.16H5 and T151). In pokeweed mitogen (PWM)-induced B cell proliferation a similar pattern of responsiveness was obtained. When we studied the anti-CD4 effects on cytokine production in T cells stimulated in mixed lymphocyte reaction (MLR) or by mitogens, we found that neither IL-2 nor IL-4 production was dramatically influenced by anti-CD4 in CD4, 45RO+ cells. This led us to the conclusion that the inhibitory effect olanti-CD4 on B cell proliferation and immunoglobulin secretion was not due lo inhibition of cytokine production. To clarify this point, we investigated the ability of anti-CD4 to inhibit conjugate formation between B and T cells. It was found that CD4, 45RO+ T cells formed more conjugates than CD4, 45RA+ cells, and that only the conjugate formation by CD4, 45RO+ T cells was inhibited by anti-CD4. These results suggest that (i) anti-CD4 inhibits T helper functions primarily by affecting CD4, 45RO+ cells, and (ii) this effect is probably mediated by contact inhibition in the early phase of T-B collaboration.
Circulating autoantibodies directed against conjugated fatty acids in sera of HIV-1-infected patients
Authors: AMARA A , CHAUGIER C , RAGNAUD J-M , GEFFARD M
Published: July 2010

Several reports have demonstrated that major changes occur in the fatty acid content of HIV-infected cells. In order to evaluate if these changes are recognized by the immune system, we have attempted to assay the possible presence of autoantibodies (autoAb) directed against conjugated fatty acids (CFA). Using an adapted ELISA, anti-CFA autoAb were assayed in sera of 150 HIV-1- infected patients and 116 controls (healthy donors and patients suffering from other diseases). Significantly increased anti-CFA autoAb of IgG class were found in HIV-1-infected patients (α < 0 001). Using our ELISA method and CFA differing in their length and their degree of unsaturation (lauric, myristic, palmitic, palmitoleic, stearic, oleic, linoienic, linolcic, ligniceric, araehidonic, eicosapentaenoic and docosabexaenoic acids), it was demonstrated that the acyl chain of CFA is the immunodominanl part recognized by these autoAb. Anti-CFA autoAb were present in 15/52 asymptomatic carriers, 14/36 symptomatic carriers, 16/39 ARC patients, but only 3/23 AIDS patients. Anti-CFA aetivity seemed to be linked with the CD4+ T cell count, and was not related to the total IgG amounts. Anti-CFA autoAb could result from self-antigen presentation to immunological cells, and may reflect lipid membrane modifications occurring in HIV-infected cells.
The significance of anti-streptokinase antibodies
Authors: LYNCH M , PENTECOST B L , LITTLER W A , STOCKLEY R A
Published: July 2010

Antibodies to streptokinase (SK) are widespread in the population, but reports of their effect on the action of SK are conflicting. Specific anti-SK IgG was purified from the sera of 10 patients, five with low titres of anti-SK IgG and five with high titres. The effect of increasing specific anti-SK IgG antibodies on the action of SK was evaluated IN vitro using a fluorimetric assay for plasmin and by a fibrin plate lysis assay. The inhibition of SK by whole plasma from a further group of patients was also assessed by the fibrin plate assay. There was a positive correlation between the serum antibody concentration and the quantity of specific anti-SK eluted (r= 0.797;P<0.005). The addition of specific anti-SK IgG caused a dose-related decrease in SK activity (fluorimetric assay r =−0.93; P = 0.02; fibrin plate assay r =−0.98: P<0.001). The addition of patient plasma to the fibrin plate assay also resulted in decreased lysis, which was dependent upon antibody titre (r=−0.95; P<0.0001). Significant in vitro reduction of the activity of SK by specific antibody was demonstrated, and this was similar with plasma containing comparable amounts of antibody. The findings suggest that treatment with SK would be unlikely to induce an effective thrombolytic slate when antibody titres are high (such as those seen within 2 years of an initial dose of SK).
IL-8 as an important chemoattractant for neutrophils in ulcerative colitis and Crohn's disease
Authors: MITSUYAMA K , TOYONAGA A , SASAKI E , WATANABE K , TATEISHI H , NISHIYAMA T , SAIKI T , IKEDA H , TSURUTA O , TANIKAWA K
Published: July 2010

IL-8 is generating increasing interest as a powerful neutrophil chemoattractant and activator. To elucidate the mechanisms of neutrophil infiltration in inflammatory bowel disease, we examined 33 patients with ulcerative colitis (UC),18 with Crohn's disease (CD), eight with some other type of colitis, and 18 normal control subjects for measurement of IL-8 in homogenates of colonic biopsy specimens. The affected colonic mucosa was found lo contain significantly more IL-8 in patients with active inflammatory bowel disease than in patients with inactive disease (UC, P<0.001: CD, P<0.001), in patients with other types of colitis (UC, P<0.05; CD, P<0.01), or in normal control subjects (UC. P<0.001; CD. P<0.001). Colonic IL-8 levels correlated significantly with the macroscopic grade of local inflammation, especially in patients with UC (P< 0.001). Colonic lL-8 levels also correlated well with the neutrophil numbers in mucosal tissue (UC, r = 0.950, P<0.001; CD. r=0-940, P<0.001), and with colonic IL-1β (r=0.911, P<0.001) and tumour necrosis factor-alpha (TNF-a) levels (r=0-604. P < 0001) in patients with these two conditions. These data suggest a potential role for IL-8 and its regulatory cytokines IL-I and TNF-a in mediating neutrophil infiltration of the gut wall in inflammatory bowel disease.
Effects of ethanol consumption and withdrawal on B cell subpopulations in murine bone marrow
Authors: KRUGER T E , JERRELLS T R
Published: July 2010

We designed studies to examine the effects of ethanol consumption and withdrawal on the numbers of pre-B and B cells in murine bone marrow. Flow cytometric analysis of B220 and surface IgM expression on bone marrow cells revealed that consumption of ethanol by mice for 7 days led to a significant reduction in pre-B cells. The number of mature B cells in the bone marrow of these animals, however, did not differ from that of control mice. In contrast, examination of bone marrow obtained from mice at various times after withdrawal from ethanol showed significantly fewer numbers of mature B cells and an even greater toss of pre-B cells. This effect was seen for relatively long periods after withdrawal. These study findings are interpreted to suggest that ethanol consumption results in changes in the pre-B cell population in murine bone marrow, II also appears that withdrawal from ethanol results in more profound changes in the mature B cell population of the bone marrow than those that occur during ethanol consumption.
Accelerated onset of collagen-induced arthritis by remote inflammation
Authors: JOOSTEN L A B , HELSEN M M A , VAN DEN BERG W B
Published: July 2010

Collagen-induced arthritis (CIA) in DBA-1 lac/J mice often has a low incidence, with gradual disease expression occurring over a broad time span (between days 35 and 70). The exact mechanisms underlying spontaneous expression are still poorly understood, although it is evident that some inflammatory cytokines like IL-1, tumour necrosis factor-alpha (TNF-α) and transforming growth factor-beta (TGF-β) are potent accelerators. We have investigated whether we could trigger collagen type II-driven inflammation by: (i) enhancing anti-collagen type II (CII) antibodies, or (ii) a non-related inflammatory process. Male DBA-1 lac/J mice were immunized with 100 μ bovine type II collagen in Freund's complete adjuvant (FCA), resulting in a low disease expression at day 28, Addition of anti-CH antibodies slightly enhanced the expression of CIA. Zymosan (3mg), given intraperitoneally, induced consistent expression of CIA after I week, whereas a retarded onset was noted with higher dosages. Local injection of a low dose of Zymosan (60μ) in the knee joint, clearly potentiated the expression of CTA at that particular site. Higher concentrations not only elicited prolonged CIA expression at the injection site, but also induced marked CIA in the draining ankle joint. In contrast, intra-articular injection of Zymosan in non-immunized DBAs or methylated bovine serum albumin (mBSA)/FCA-immunized controls only induced transient joint inflammation. The nature of the highly erosive CIA was confirmed histologically, and could easily be discriminated from the reversible changes induced with Zymosan. Our data indicate that latent autoimmune reactions may come to expression at the moment of non-specific inflammation, even at a remote site.
Immunohistological assessment of cutaneous drug hypersensitivity in patients with HIV infection
Authors: CARR A , VASAK E , MUNRO V , PENNY R , COOPER D A
Published: July 2010

The pathogenesis of drug hypersensitivity in patients with HIV infection is unknown. To study further the nature of hypersensitivity. The histopathological features of morbilliform drug hypersensitivity reactions were examined in a group of HIV-infected patients. Skin sections from 23 HIV-infected subjects with morbilliform drug hypersensitivity reactions were examined by light microscopy, direct immunofluorescence and immunohistochemistry, to determine the nature of the inflammatory infiltrate and the role of immunoglobulin. complement and cytokines. The principal light microscopic findings were spongiosis, hydropic generation of the basal layer, civatte bodies, an epidermal Iymphocytic infiltrate (48%). and a perivascular dermal infiltrate of lymphocytes (87%) and macrophages (52%). Two patients had findings consistent with toxic epidermal necrolysis. Immunohistochemistry demonstrated that the lymphocytic infiltrate consisted of CDS+. HLA-DR+ T lymphocytes (some of which also stained for CD38), a marked depletion of epidermal Langerhans cells (90%), and strong cytoplasmic staining of keratinocytes for IL-6 (60%), IL-1β(50%). tumour necrosis factor-alpha (TNF-α) (45%) and to a lesser degree, interferon-gamma (IFN-7) (35%). Immunofluorescence did not demonstrate any significant deposition of immunoglobulin or complement. The histological findings were independent of there responsible drug, the duration of either therapy or the rash, and of peripheral blood CD4+ and CD8+ cell counts. These findings suggest that activated CD8+ lymphocytes and perhaps epidermal production of cytokines are involved in the pathogenesis of cutaneous drug hypersensitivity in HIV-infected patients. The common histological features, regardless of the causative drug, suggest a common pathogenesis.
The role of natural killer cells in resistance to coccidiosis: investigations in a murine model
Authors: SMITH A L , ROSE M E , WAKELIN D
Published: July 2010

Natural killer (NK) activity, detected by the lysis of Yac-1 target cells, was examined in splenic and mesenteric lymph node (MLN) cells throughout the course of infection with Eimeria vermiformis in BALB/c and C57BI/6 (B6) mice. These strains are, respectively, relatively resistant and susceptible to primary infections, which render them equally, and completely, resistant to challenge. Resting levels of NK activity were higher in B6 than in BALB/c, and B6 responded earlier in the course of infection than BALB/c, but splenic peak values were higher in BALB/c; the pattern of response in MLN cells was similar in both strains, but the peak was higher in BALB/c. At the lime (7 days p.i.) of peak NK response in BALB/c mice there was, depending upon the choice of NK-resistant/lymphokine-activated killer (LAK)-sensitive target cells, either little (P388D1), or no (P815) splenic LAK activity. Challenge of immunized BALB/c mice did not evoke a detectable NK response. Although the higher NK activity in BALB/c mice correlated with greater control of primary infection, depletion of NK activity (demonstrated in splenic cells) in vivo by treatment with anti-asialo GM1 antibodies did not greatly affect the course of infection. Furthermore, this treatment did not augment the exacerbation of infection produced by treatment with antiinterferon-gamma (IFN-γ) MoAb, indicating that, at least in this system, NK cells are not a fundamentally important source of this controlling cytokine of eimerian infections. The results suggest that NK cells may not greatly influence the outcome of coccidial infections.
Reduced NK activity correlates with active disease in HIV- patients with multidrug-resistant pulmonary tuberculosis
Authors: RATCLIFFE L T , LUKEY P T , MACKENZIE C R , RESS S R
Published: July 2010

There has been a global increase in the incidence of multidrug-resistant pulmonary tuberculosis (TB). As there are no previous reports of immune function in HIV- patients with multidrug-resistant pulmonary TB, a comprehensive assessment of cellular immunity in this setting was undertaken. This involved a prospective, case-controlled study which included five patients with active multidrug-resistant pulmonary TB and five matched controls with active non-resistant infection, and documented the changes in immune parameters which occurred upon clinical resolution. Patients with multidrug-resistant TB had significantly lower fresh natural killer (NK) cell activity than matched controls with non-resistant pulmonary TB (P < 0.05). This was a specific abnormality, as there were no significant differences in antigen-specific cytotoxicity or lymphocyte proliferation in the case-controlled study. Follow-up assessment of the patients with multidrug-resistant infections indicated that clinical improvement correlated with a moderate increase in NK cell activity. Impaired NK cell function may be involved in the pathogenesis of multidrug-resistant TB.
Effects of decomplementation with cobra venom factor on experimental vasculitis
Authors: MATHIESON P W , QASIM F J , THIRU S , OLDROYD R G , OLIVEIRA D B G
Published: July 2010

Mercuric chloride (HgCl2) induces autoimmunity in susceptible rat strains, with hyper-IgE, appearance of a number of autoantibodies and wide spread tissue injury, including necrotizing vasculitis in the gut. In the early phase of tissue injury there is granulocyte infiltration; later there is immunoglobulin deposition along basement membranes in vessels. We have analysed the role of complement in this model using cobra venom factor (CVF). which causes decomplementation lasting around 5 days. The characteristic time course when HgCl2 is given over 10 days is that tissue injury and autoantibody levels reach a peak at around day 15 (start of HgCl2= day 0). We therefore gave CVF either early (day 0). intermediate (day 5) or late (day 10); a fourth group (controls) received HgCl2 but no CVF. At each time point, CVF caused complete decomplementation which lasted for al least 5 days, Serum IgE and autoantibody levels were similar in all four experimental groups. Tissue injury in the “early” CVF group and in the ‘Jale’ CVF group was not significantly different from control, but in the intermediate group tissue injury was significantly more severe than in controls. These data indicate that the complement system does not play a major rote in the induction of autoantibodies by HgCl2, nor in the effector phase of tissue injury. We speculate that the exacerbation of tissue injury by CVF in the group given this agent al an intermediate stage of the model is explained by the presence of products of C3 activation which have proinflammatory effects during the phase of active granulocyte-mediated tissue injury.
Possible association of CD3 and CD4 polymorphisms with insulin-dependent diabetes mellitus (IDDM)
Authors: GHABANBASANI M ZAMANI , BUYSE I , LEGIUS E , DECORTE R , MARYNEN P , BOUILLON R , CASSIMAN J-J
Published: July 2010

Population and family studies show that predisposition to type I diabetes (IDDM) is multi-factorial, and that polymorphisms in the MHC region contribute substantially to the susceptibility to IDDM. In the present study the association of polymorphisms in the CD4 and the 6 subunit of CD3 with IDDM were examined in a Belgian population. We observed that the frequency of the CD4*A4/A4 genotype and of the CD3*91 allele were significantly increased (P= 0·0077) and decreased (P = 3·8 × 10−5), respectively, in IDDM compared with controls. These results therefore suggest that CD4, CD3 or neighbouring genes might contribute to IDDM susceptibility. These results are, however, preliminary and cannot be considered as established until re-tested in a new population.
Expression and functional role of 1F7 (CD26) antigen on peripheral blood and synovial fluid T cells in rheumatoid arthritis patients
Authors: MUSCAT C , BERTOTTO A , AGEA E , BISTONI O , ERCOLANI R , TOGNELLINI R , SPINOZZI F , CESAROTTI M , GERLI R
Published: July 2010

The expression and the functional role of the CD26 (1F7) T cell surface molecule, an ectoenzyme which seems to represent a functional collagen receptor of T lymphocytes and to have a role in T cell activation, were analysed in both peripheral blood (PB) and synovial fluid (SF) T cell samples from patients with active and inactive rheumatoid arthritis (RA). Although patients with active disease displayed higher percentages of PB CD26+ CD4+ T cells than inactive RA and control subjects, CD26 antigen expression on RA SF T lymphocytes was low. The anti-1F7 binding to the T cell surface, that led to CD26 antigen modulation and enhancement of both IL-2 synthesis by, and 3H-TdR incorporation of, anti-CD3- or anti-CD2-triggered PB T cells in RA and control subjects, was unable to affect significantly both expression and functional activity of RA SF T lymphocytes. Since the 1F7 antigen spontaneously reappeared on the surface of unstimulated SFT cells after 2-5 days of culturing, the low IF7 antigen expression of anti-lF7 in the SF T cell compartment may be the result of in vivo molecule modulation exerted by the natural ligand in the joint, with important implications for T cell activation and lymphokine synthesis.
The influence of anti-fibronectin antibodies on interactions involving extracellular matrix components and cells: a possible pathogenic mechanism
Authors: ATTA M S , POWELL R J , TODD I
Published: July 2010

Antibodies, directed to the 30-kD collagen binding domain (CBD) of fibronectin (Fn), have been previously demonstrated in sera from patients with systemic lupus erythematosus (SLE), and we now investigate the possible pathogenic effects of these antibodies on collagen-Fn and cell-Fn interactions. The binding of type 1 collagen to Fn was demonstrated by ELISA, and could be specifically inhibited by the preincubation of solid-phase immobilized Fn with anti-Fn antibodies from SLE sera. By using indirect immunofluorescent staining, anti-Fn antibody containing SLE sera but not normal human serum (NHS) reduced the deposition of newly synthesized collagen and Fn on living human skin fibroblasts. We also found that sera from SLE patients containing anti-Fn antibodies significantly reduced thyroid cell attachment to Fn immobilized on plastic compared with NHS. These effects were shown to be due to the presence of anti-Fn antibodies in these sera, as SLE sera depleted of anti-Fn antibodies did not reduce the deposition of collagen or Fn on cultured fibroblasts, nor did they inhibit cell attachment.
Elevation of serum soluble intercellular adhesion molecule-1 (sICAM-1) and sE-selectin levels in bronchial asthma
Authors: KOBAYASHI T , HASHIMOTO S , IMAI K , AMEMIYA E , YAMAGUCHI M , YACHI A , HORIE T
Published: July 2010

Adhesion molecules such as ICAM-1 and E-selection have been shown to play important roles in the production of allergic inflammation. In the present study, we measured serum soluble ICAM-1 (sICAM-1)and soluble E-selection (sE-selectin) levels by ELISA in 42 patients with bronchial asthma (22 atopic and 20 non-atopic) during asthma attacks and in stable conditions in order to assess the state of ICAM-1 and E-selectin in allergic inflammation. Both serum sICAM-1 levels and serum sE-sclectin levels in sera obtained during bronchial asthma attacks were higher than those in sera obtained in stable conditions. These findings were observed regardless of atopic status. To examine the regulatory mechanism in the elevation of serum sICAM-1 and sE-selectin levels, serum tumour necrosis factor-alpha (TNF-α) levels were measured by ELISA. TNF-α levels in sera obtained during bronchial asthma attacks were higher than those in sera obtained in stable conditions. There was a correlation between the nature of change in serum TNF-α levels and the nature of change in serum slCAM-1 levels or serum sE-selectin levels, though serum TNF-α levels did not correlate with serum sICAM-1 levels or serum sE-selectin levels. These results suggest that higher levels of sICAM-l and sE-selectin during asthma attacks may reflect the up-regulation of ICAM-1 and E-selectin expression in allergic inflammation, and that the soluble form of these adhesion molecules may be useful markers for the presence of allergic inflammation. TNF-α is shown to enhance the expression and release of ICAM-1 and E-selectin in vitro, however; the regulatory mechanism in the elevation of serum sICAM-1 and sE-selectin levels remains to be clarified.