bluuebunny/crossref_metadata_2025 huggingface dataset:
The effects of recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-3 on the secretory capacity of human blood eosinophils
Authors: TAI P C , SPRY C J
Published: July 2010

The effects of recombinant human GM-CSF and interleukin-3 (IL-3) on human blood eosinophil survival, activation, and secretion were studied. Purified normal density eosinophils from patients with the idiopathic hypercosinophilic syndrome (HES) survived in culture for 7 days (50% viable) in the presence of 50 nM GM-CSF or 50 nM IL-3. Ncutrophils did not survive after 4 days. No eosinophils survived in the absence of GM-CSF or IL-3. In two out of five patients studied, the cultured eosinophils became elongated with numerous processes. In all five patients the cells became adherent, but there were no morphological signs of degranulation. Both GM-CSF and IL-3 activated eosinophils, transforming the storage form of eosinophil cationic protein (ECP) into the secreted form. The proportion of activated cells increased from less than 20% to over 50% after 4 days in culture. However, GM-CSF and IL-3 did not induce secretion on their own. On the other hand, when GM-CSF/IL-3-activated eosinophils were exposed to known secretory stimuli, there was a six-fold increase in the amount of ECP released when the cells were stimulated with sepharose coated with C3b. and a two-fold increase when they were stimulated with sepharose-activatcd whole autologous serum. Eosinophils from patients taking steroids were unable to secrete their granule contents, even though they became activated by GM-CSF and IL-3. A novel finding was that sepharose-activated whole serum was an extremely potent secretory signal for ECP. releasing up to 50% of the total ECP content. These studies showed that GM-CSF and IL-3 prime eosinophil effector function by initiating granule solubilization which is the first step in the secretory event, without affecting the subsequent extracellular release of granule proteins.
CD4+ cytolytic T cell clones that recognize polymorphism of HLA-DRβ3 chains
Authors: NOGUCHI M , HOZUMI N , NISBET-BROWN E
Published: July 2010

We have investigated HLA-DRβ3-associated functional polymorphism using selected Epstein-Barr virus (EBV) specific human T cell clones and EBV-transformed B cell (EBV-B) lines. To study the relationship between T cell recognition and the gene products of the three alleles of the DRβ3 locus, Dw24, 25 and 26 (these were previously called DRw52a, b and c, respectively), CD4+ cytolytic T cell clones (CD4+ CTL) were isolated by repeated stimulation of peripheral blood mononuclear cells (HLA A2 A24; B8 B27; DRw17, Dw24, DRw2) with autologous EBV-B. Clone no. 32 proliferated strongly in response to HLA-Dw24 EBV-B, but not to Dw25 or Dw26 EBV-B. Furthermore, clones no. 32 and no. 45 both lysed HLA-Dw24 EBV-B but not Dw25 or Dw26 EBV-B. In addition, cold target inhibition studies showed that the cytolytic activity of both clones was blocked by unlabelled HLA-Dw24 EBV-B, but not by Dw25 or Dw26 EBV-B. Clones no. 32 and no. 45, therefore, could distinguish between the three allelic products of DRβ3 haplotypes.
Thymus lymphocytes in uraemic rats and the effect of thymosin fraction 5 in vitro
Authors: IKEMOTO S , KISHIMOTO T , WADA S , HAYAHARA N , MAEKAWA M
Published: July 2010

Cellular immunity of thymus lymphocytes in uraemic rats was studied. Severe and moderate uraemia was induced in rats, and sham-operated and normal rats were used as the controls. As a result, the response of thymus lymphocytes to concanavalin A (Con A) significantly decreased in severely uraemic rats, but did not change in moderately uraemic rats. However, when the thymus lymphocytes were pretreated with thymosin fraction 5, the response to Con A was ameliorated in severely uraemic rats. There was a significant correlation between the effect of thymosin fraction 5 on Con A response and Con A response of thymus lymphocytes. In addition, serum from severe uraemic rats suppressed the response of normal thymus lymphocytes to Con A. These results indicate that severe uraemia may cause an impairment in maturation of thymus lymphocytes, which can be improved by thymosin fraction 5 in vitro.
Cloning of human T cells specific for measles virus haemagglutinin and nucleocapsid
Authors: ILONEN J , MÅKELÅ M J , ZIOLA B , SALMI A A
Published: July 2010

T cell lines specific for measles virus (MV) were generated from blood of two DR1/DR2 heterozygous healthy donors with a history of past measles infection. The antigenic specificity of 66 T cell clones derived from the lines was studied in a blastogenic assay using whole measles virus and two purified virus components, haemagglutinin and nucleocapsid. Thirty-nine of the clones were specific for one of the two purified antigens. None of seven synthetic peptides covering 20% of the MV haemagglutinin amino acid sequence stimulated T cell clones with haemagglutinin specificity. Responsiveness of the majority of the clones were restricted by HLA-D/DR antigens, although two clones were isolated that responded only to MV antigens presented by autologous cells. Ten of 11 clones recognizing the nucleocapsid antigen were DR1-restricted, while the haemagglutinin antigen and whole measles virions were recognized more often in association with the DR2 antigen. These results indicate that much of the MV-specific memory T cell response is specific for the haemagglutinin and nucleocapsid virus antigens, with the DR antigen being the main restriction element involved.
The development of primed/memory CD8+ lymphocytesin vitroand in rejecting kidneys after transplantation
Authors: AKBAR A N , AMLOT P L , TIMMS A , LOMBARDI G , LECHLER R , JANOSSY G
Published: July 2010

In normal individuals, 80 ±5% of circulating CD8+ T cells express CD45RA, and 20 ± 7% of these cells express CD45R0 antigens. After activation, CD8+ cells expressing CD45RA decrease to 56–67% while those expressing CD45R0 increase to 38–67%. Although precursors of alloantigen-specific cytotoxic T cells were found in both CD8+, CD45RA+ and CDS+, CD45R0+ subsets, the specific effector cells were exclusively CD8+, CD45R0+, Allospecific cytotoxic CD8+ clones were also entirely CD45R0+, A lectin-dependent cytotoxic (LDC) assay unmasked a hierarchy of killing after alloactivation which was CD8+, CD45R0+ > CD8+, CD45RA+ > CD4+, CD45R0+ > CD4+, CD45RA+, The phenotype of CDS+ T cells in rejecting kidneys was similar to in vitro alloantigen-activated CDS+, CD45R0+ cells and cytotoxic CD8+ clones. Firstly there was an increase in the relative proportions of CD45R0+ (60·8%) and a decrease in CD45RA+ (35·10%) CD8+ cells relative to circulating CD8+ subsets. In the rejecting grafts. 34 ± 9% of the CD8+ cells were also DR+ indicative of recent activation. Furthermore, 16% (range 4–35%) of rejecting CD8+ cells were Ki67+ suggesting that these cells were proliferating. Finally, 17% (range 4–53%) of T cells in rejecting kidneys simultaneously expressed both CD45RA and CD45R0 markers. These results show that in vitro alloantigen-activated CD8+, CD45R0+ cells represent a primed/memory cytotoxic population. In addition, they provide indirect evidence that a proportion of CD8+ cells in rejecting kidneys were actively switching from a naive to a memory phenotype in vivo in a manner analogous to that in vitro.
Pneumonitis in bone marrow transplant recipients results from a local immune response
Authors: MILBURN H J , BOIS R M DU , PRENTICE H G , POULTER L W
Published: July 2010

Eighteen recipients of allogeneic T cell-depleted bone marrow who developed 22 episodes of interstitial pneumonitis were investigated by bronchoalveolar lavage for the cause of pneumonitis. The cells obtained were examined using a panel of monoclonal antibodies with immunocytochemical techniques to identify lymphocyte subsets and the presence of surface molecules indicative of lymphocyte activation. The majority of patients had an excess of lymphocytes in lavage and most of these cells were positively stained with the McAb recognizing the CD8 antigen (suppressor/cytotoxic type T cells). Although the proportions of CD4+ (helper type) T cells were below normal, the absolute numbers were within normal limits, thus the CD4:CD8 ratio was consistently 1:1 or less. A large proportion of the CD8+ cells displayed HLA-DR molecules (RFDR1+), interleukin-2 (IL-2) receptors (CD25+) and high concentration of CD7 antigen (RFT2+). Further analysis revealed that most CD8+ cells were CD5+ (RFT1+) yet a large proportion (20–40%) were CD5−. A majority of CD8+ cells was also CD38+ (RFT10+) and Leu7+. No clear correlation between the emergence of a raised proportion of activated CD8+ cells and diagnosed cytomegalovirus infection was found. These results demonstrate, however, that cells with the phenotype of the resident T cells of the bronchial epithelium (CD8+CD5−) emerge to the air spaces and express activation markers. This raises the intriguing paradox of an aggressive local immune response occurring in an otherwise immunosuppressed group of patients.
Rheumatoid factors and glomerulonephritis
Authors: MIYAZAKI M , ENDOH M , SUGA T , YANO N , KURAMOTO T , MATSUMOTO Y , EGUCHI K , YAGAME M , MIURA M , NOMOTO Y , SAKAI H
Published: July 2010

It is presently unknown whether rheumatoid factors have a pathogenic role in the development of various types of glomerulonephritis with immune deposits. Three isotypes of rheumatoid factors (RFs), which are autoantibodies to IgG, were measured using the solid-phase fluorescence immunoassay in sera from patients with diffuse proliferative lupus nephritis (DPLN), membranous lupus nephritis (MLN), IgA nephropathy (IgAN) and idiopathic membranous nephropathy (MN). RF activity of immunoglobulins deposited in the glomeruli from these patients was also studied by examining the binding of the FITC-conjugated human IgG and Fc portion of IgG to the glomeruli of renal biopsy specimens. IgG, IgA and IgM RFs were significantly increased in sera from patients with DPLN. and the increase was significantly lower in patients with MLN. IgAN and MN. Human IgG bound to immunoglobulin on the glomeruli only in DPLN, but not in MLN, IgAN or MN. The Fc portion of IgG was demonstrated to be involved in this reaction. It was suggested that RFs and IgG may play a major role in immune deposits on the glomeruli in DPLN and may be involved in the development of DPLN; however, this is not likely in MLN, IgAN or MN.
Tumour necrosis factor-alpha and interferon-gamma production measured at the single cell level in normal and inflamed human intestine
Authors: MACDONALD T T , HUTCHINGS P , CHOY M-Y , MURCH S , COOKE A
Published: July 2010

The spot-ELISA technique has been used to enumerate the frequency of cells secreting tumor necrosis factor-alpha (TNF-α) and interferon-γ (IFN-γ), isolated from biopsies of normal intestine and from biopsies of children with inflammatory bowel disease. TNF-α production was undetectable in six out of 12 biopsies from normal intestine and in the other six biopsies it ranged from 60 to 580 TNF-α-secreting cells/106 isolated intestinal cells. In contrast, cells isolated from biopsies of children with Crohn's disease (n= 9) all showed elevated frequencies of TNF-á-secreting cells (500–12 000 secreting cells/106 cells). In ulcerative colitis, four out of eight children had increased production of TNF-α and in children with indeterminate colitis two out of three had elevated levels. There was no correlation between plasma TNF-α levels and the number of intestinal cells secreting TNF-α. In controls and all groups of patients IFN-γ-secreting cells were uncommon. These results suggest that TNF-α is an important mediator of inflammation in the human gut, and, furthermore, may play a role in the growth failure frequently seen in children with inflammatory bowel disease.
Influence of the G2m(n) allotype and age on IgG subclass distribution in antibodies to dietary proteins in children with coeliac disease
Authors: RAUTONEN N , RAUTONEN J , SAVILAHTI E
Published: July 2010

In 47 children with coeliac disease, IgG 1 was the dominant IgG subclass (mean 61 %) in antibodies to gliadin, a protein component of wheat; IgG2 (22%) and IgG3 (15%) were also found. Very little IgG4 (3%) was detected. Both G2m(n)allotype and the age of the patient had an independent effect on the subclass distribution. The effects of these two factors seem similar to those described earlier on responses to polysaccharides; both G2m(n) and age increase the expression of IgG2 antibodies and concomitantly the share of IgGl is decreased. The average increase in the share of IgG2 brought about by n/n-genotype was 25-3% (95% confidence limits 12.2–38.4%; P<0.001) and that brought about by each additional year of age of the patient 11% (0.3–1.9%;P=0.006). IgG antibodies to /Mactoglobulin and ovalbumin consisted mainly of IgGl and IgG4. The titres of the IgG subclasses were low and in many cases below detection level. No association with G2m(n) genotype and share of IgGl or IgG4 could be demonstrated. Age was inversely correlated with the shares of IgGl in antibodies to β-Mactoglobulin and ovalbumin.
Enhanced expression of the tumour necrosis factor/cachectin gene in peripheral blood mononuclear cells from patients with systemic vasculitis
Authors: DEGUCHI Y , SHIBATA N , KISHIMOTO S
Published: July 2010

The expression of tumour necrosis factor-alpha (TNF-α) gene in peripheral blood mononuclear cells from patients with systemic vasculitis. periarteritis nodosa and Wegener's granulomatosis, has been studied by RNA dot blot and Northern blot assays. We further examined the transcriptional level of TNF-α gene in peripheral blood mononuclear cells from these patients by performing nuclear run on transcription assay. We demonstrate enhanced TNF-α gene expression in mononuclear cells from these patients compared with healthy subjects and patients with bronchial asthma. These findings suggest that the enhanced transcription of TNF-α gene in peripheral blood mononuclear cells from patients with systemic vasculitis may be involved in the pathophysiology/pathogenesis of these diseases by cylokine dysregulation.
Stimulation of neutrophil elastase and myeloperoxidase release by IgG fragments
Authors: ECKLE I , KOLB G , HEISER C , HAVEMANN K
Published: July 2010

Human leucocyte elastase (HLE) cleaves IgG into Fab and Fc fragments. The Fc fragment bears an elastase-specific antigen and has previously been reported to be found in synovial fluid during rheumatoid arthritis, In addition, biological activity of clastasc-specific Fc fragments has been described in modulating granulocyte oxidative metabolism. To investigate further regulatory effects of the elastase-induced IgG cleavage products, we tested the elaslase and myeloperoxidase release of granulocytes. IgG fragments induce no enzyme release of unstimulated neutrophils. But elastase and myeloperoxidase release of cytochalasin b/FMLP-treated neutrophils is stimulated in a dose-dependent manner by the Fab fragments. The extent of stimulation depends on stimulus concentration and is at its maximum for low (e.g. 2.5 × 10−8m) FMLPconcentration. Ten nanomoles Fab/4 × 106 PMN augment elastase release to 206% and myeloperoxidasc release to 155% after prestimulation with 2.5 × 10−8 m FMLP. Fc fragments stimulate elaslasc release to 162% but no MPO release. Untreated IgGl and analog Fab and Fc fragments produced by papain cleavage react similarly. Elastase-generated IgG fragments may therefore up-regulate their concentration by stimulating elastase release. The concomitantly stimulated release of myeloperoxidase may influence bactericidal activity and termination of oxidative burst.
A complex component modulating immune-deficient cells in leprosy patients leading to loss of viability ofMycobacterium leprae—a possible vaccine
Authors: MAROLIA J , ROBINSON P , MAHADEVAN P R
Published: July 2010

Macrophages from peripheral blood of leprosy patients, both multi-bacillary and paucibacillary are unable to kill phagocytosed Mycobacterium leprae due to their inability to produce superoxide (O2−) and hydroxyl radicals (OH ·). The macrophages from healthy individuals are able to kill M. leprae along with release of O2− and OH · radicals. The deficiency in the macrophages of both types of leprosy patients is removed by activation of these cells when exposed to a culture supernatant obtained after stimulation of peripheral blood mononuclear cells from the same patients with delipidified cell components of M. leprae which are most likely cell wall proteins. The activation of macrophages also leads to recognition of whole live M. leprae as an antigen by cells from lepromatous patients. This activation of the phagocytes by delipidified cell components is blocked by cyclosporin A, indicating the possible role of several steps involved in immune activation of cells. The observations thus indicate the significant ability of delipidified cell components to eliminate the deficiencies in the macrophages from leprosy patients and restore them to behave like the cells from healthy individuals. Considering all these, it is suggested that delipidified cell components could be potential modulators, and are probably capable of functioning as a vaccine for leprosy.
Anti-myeloperoxidase antibodies in systemic vasculitis
Authors: LEE S S , ADU D , THOMPSON R A
Published: July 2010

Anti-neutrophil cytoplasmic antibodies (ANCA) are important diagnostic markers in vasculitic disorders. In a study of 164 patients with various clinical syndromes associated with vasculitis, 60 were found to have the antibody, as detected by indirect immunofluorescence; 23 had the so-called perinuclear antibody pattern, and of these, 15 had antibodies to neutrophil myeloperoxidase in an ELISA. These patients had multi-system involvement, and 14 of the 15 had histological evidence of small-vessel arteritis in the kidneys. In preliminary experiments, the isolated IgG from one patient was shown to inhibit luminol-dependent chemiluminescence of fluid phase myeloperoxidase.
Liver membrane antibodies (LMA) recognize a 26-kD protein onthe hepatocellular surface
Authors: HOPF U , JAHN H-U , MÖLLER B , STEMEROWICZ R , WITTENBRINK C , KLEIN R , BERG P A
Published: July 2010

Sera from 82 patients with chronic inflammatory liver diseases and from patients with systemic lupuserythemalosus (SLE). rheumatoid arthritis (RA) and Hashimoto's thyroiditis were studied byimmunoblotting against purified liver plasma membranes (LPM) and soluble liver protein (SLP)fractions from different species after previous separation by SDS-PAGE. Eighteen of 19 sera withLMA of IgG type in immunofluorescencc assay and six LMA-negatiuve sera (three sera from patientswith RA) showed antibodies of the IgG or IgM classes against a protein with a molecular weight of 26kD which was present in LPM and SLP fractions from rats, rabbits, pigs and humans. The reactionwith 26-kD liver protein did not correlate with other known autoantibody-antigen systems. All serawere negative in the 26-kD region with liver mitochondria, liver microsomes and soluble proteins ofkidney (with one exception), heart and gut from the rat. The 26-kD protein was purified by affinitychromatography on immobilized anti-26-kD protein antibodies from patients, eluted from the 26-kDband of immunoblots. Studies with purified 26-kD liver protein and with SLP as antigens afterseparation in two-dimensional electrophoresis confirmed that patient serum and experimental rabbitantiserum react with the same protein. Eluted patient antibodies and rabbit antisera showed a linearfluorescence pattern on isolated hepatocytes from rat and rabbit. The data indicate that one of thetarget antigens of LMA is a species-nonspecific 26-kD protein located on the hepalocellular surface.
Tumour necrosis factor production and cell-mediated immunityin anorexia nervosa
Authors: SCHATTNER A , STEINBOCK M , TEPPER R , SCHONFELD A , VAISMAN N , HAHN T
Published: July 2010

Fourteen patients with anorexia nervosa (AN) were studied for the production of tumour necrosisfactor (TNF), the activation of the interferon (IFN) system and cell-mediated cytotoxicity (CMC)and the results were compared with 16 age-matched healthy women. AN patients had significantlyincreased spontaneous TNF production by peripheral blood mononuclear cells (PBMC) in vitro(16 ± 5 U/ml versus 4 ± 3 U/ml in the control group; P < 005), although no TNF was detectable in theplasma from either group. TNF production in vitro, following stimulation of PBMC byphytohaemagglutinin (PHA) or tumour cells, was similar in AN patients and controls; however, lipopolysaccharide (LPS) induced TNF production was found to be lower in AN (P <0.1). CMC wassignificantly lower in AN patients (4 ± 2 versus 10 ± 3 in controls, expressed as lytic units/106 cells;P<0.05), but no difference could be found between AN and controls in IFN activity as reflected bythe level of the IFN-induced enzyme 2′-5′oligoadenylate synthetase (2-5A) in PBMC. Beta-endorphins in the plasma were higher in the AN group (P < 0.05) but these levels could not becorrelated to those of lFN, CMC or TNF. Defective CMC and increased TNF production by PBMCin patients with anorexia nervosa may possibly result from the nutritional deficiencies andneuroendocrine abnormalities associated with the disease, and may contribute to the pathophysiology of AN.
Pathogenesis of experimental lupus nephritis: a role foranti-basement membrane and anti-tubular brush border antibodiesin murine chronic graft-versus-hostdisease
Authors: BRUIJN J A , HOGENDOORN P C W , CORVER W E , VAN DEN BROEK L J C M , HOEDEMAEKER P J , FLEUREN G J
Published: July 2010

The pathogenesis of renal involvement was studied in murine chronic graft-versus-host disease(GVHD), which is a model for human systemic lupus erythematosus. GVHD was induced by Tour i.v.injections of lymphocytes from DBA/2 donor mice into (C57BL/10 x DBA/2)Fi hybrids at 3-4-dayintervals. Two weeks after the first injection, antibodies were found to have been deposited in themesangium and along the glomerular basement membrane (GBM) in a linear arrangement, whichchanged to a granular pattern after 6–8 weeks. In this stage, large electron-dense complexes werepresent both subepithelially and subendothelially along the GBM. Proteinuria increased up to11 300 ± 2140 μg/ 18 h. Indirect immunofluorescence studies and ELISA showed that sera and kidneyeluates contained autoanlibodies directed against nuclear antigens and GBM component laminin aswell as against renal tubular epithelial antigens (RTE). The specificity of the anti-RTE antibodies wasfurther characterized by the use of absorption techniques as well as immunoblotting. The early linearimmunofluorescence pattern seems to be associated with glomerular binding of anti-GBMantibodies, while electron-dense complex formation in later stages may be induced by thesuperimposed deposition of anti-RTE antibodies. Similar phenomena were recently described inHeymann's nephritis in the rat, a model for human membranous nephropathy.
Hashimoto's thyroiditis manifesting monoclonal lymphocytic infiltration
Authors: MATSUBAYASHI S , TAMAI H , MORITA T , FUKATA S , MATSUZUKA F , SUZUKI T , KUMA K , NAGATAKI S , VOLPÉ R
Published: July 2010

Hashimoto's thyroiditis (HT) and lymphoma arc sometimes difficult to distinguish between. Moreover, lymphoma sometimes develops in a thyroid gland from pre-existing HT. Open- or large-needle biopsy usually distinguishes between them; the specimen may be examined histologically and subjected to immunohistochemistry. Another possible method of examination is fine-needle aspiration biopsy (FNAB). The cells obtained may be evaluated cytologically, and subjected to flow cytometry, using various antibodies. In this study, anti-kappa and anti-lambda antibodies are especially important, as a gross predominance of kappa or lambda B lymphocytes infiltrating the thyroid is evidence for a B cell monoclone. In this study, 15 patients were selected because of their rapidly growing goitres. They all underwent FNAB. Five had cytology typical of HT, and no evidence of monoclonality on flow cytometry. They were diagnosed as HT without further histopathology. The remaining 10 patients had cytology suspected of lymphoma, or evidence of monoclonality on flow cytometry, or both. These patients underwent open- or large-needle biopsy. Only three of them were diagnosed histopathologically as lymphoma; the other seven were diagnosed histopathologically as HT, making 12 cases of HT in all. Five of these 12 cases, and one of the three cases of lymphoma showed flow cytometrical evidence of monoclonality; thus evidence of monoclonality from FNAB, while interesting, does not necessarily serve to differentiate between HT and lymphoma. Furthermore, the immunohistochemical assessment of monoclonalily did not correlate with the flow cytometrical assessment. Follow-up evidence will be required to discover whether those patients with a B cell monoclone in their HT are the ones who develop a lymphoma.
The effect of cyclosporin A on the interstitial mononuclear cell infiltration and the induction of Heymann's nephritis
Authors: GRÖNHAGEN-RISKA C , VON WILLEBRAND E , TIKKANEN T , HONKANEN E , MIETTINEN A , HOLTHÖFER H , TÖRNROTH T
Published: July 2010

Heymann's nephritis was induced with brush-border (BB) antigen. Interstitial mononuclear cell infiltration was studied with cytological examinations of fine-needle aspiration biopsies (FNAB), and with immunoperoxidase stains of frozen sections with monoclonal antisera. The effect of cyclosporin A (CyA), 20 mg/kg when administered intraperitoneally for 8 days in association with both initial immunization, and with the booster 4 weeks later, on the interstitial leukocyte infiltration and on the development of membranous glomerulonephritis (MGN) and proteinuria were investigated. Another group of rats was immunized, but not given CyA. Experimental animals were killed in groups 3, 6 and 20 weeks after initial immunization. CyA inhibited significantly the initial interstitial lymphocyte and blast cell response at 3 weeks (FNAB), but did not inhibit the secondary response after the booster. The anti-BB titre reacted in a similar fashion. Immunoperoxidase stains indicated a clearly suppressed T suppressor/cytolytic (T s/c) cell response. Glomerular basement membrane (GBM) deposits of IgG developed more slowly and were more scarce in the CyA-treated rats, when compared with the untreated group. Only one out of 15 CyA treated rats developed C3 deposits in the GBM during the course of the study, and none developed proteinuria, when most untreated rats (10/ 17) had C3 deposits and were nephrotic at 20 weeks. Thus, CyA depressed the initial interstitial cellular response after immunization with BB antigen, and also inhibited the development of antibody response, C3 deposits and proteinuria of Heymann nephritis. These effects of CyA may be contributed to an inhibited amplification of the autoimmune response associated with interstitial damage and continuous release of autoantigen.
Role of antigen-presenting cells in variation in immunogenicity of mycobacteria
Authors: SHROFF K E , SAINIS K B , SENGUPTA S R , KAMAT R S
Published: July 2010

The route of immunization was observed to play a significant role in deciding the outcome of immunization with killed mycobacterial vaccines. Whereas the slow growers were immunogenic by both intraperitoneal and intradermal routes, the rapid growers were immunogenic only by intradermal route. The non-responder state of mice to Mycobacterium vaccae by i.p. route of immunization could be corrected by prior treatment with poly I:poly C, an interferon inducer, or indomethacin, a prostaglandin inhibitor. Antigen-presenting efficiency of peritoneal and spleen cells were compared employing M. vaccae and M. tuberculosis H37Rv primed T cells and corresponding sonicates as antigens in an in vitro lymphocyte transformation test. Irradiated spleen cells presented both the antigens efficiently. However, with peritoneal cells as antigen-presenting cells, proliferative response against only M. tuberculosis was observed; proliferation of M. vaccae primed T cells was very poor. Peritoneal cells of poly I:poly C treated mice showed distinct improvement in their efficiency of presentation; even paraformaldehyde-fixed peritoneal cells gave an efficient stimulation with M. vaccae. The percentage of la-positive fraction in peritoneal cells was very low (5.95%) in comparison with spleen cells (38.37%). Poly I: poly C treatment resulted in increase in the la-positive cell fraction of the peritoneal cells to 24.5%.
Requirement of the T cell receptor for antigen presentation by T lymphocytes. Effect of envelope glycoproteins of HIV-1 on antigen presentation by T cells
Authors: CHIRMULE N , KALYANARAMAN V S , SLADE H , OYAIZU N , PAHWA S
Published: July 2010

We have developed CD4+, tetanus antigen-specific T cell clones that proliferate in the presence of tetanus antigen and autologous irradiated peripheral blood leucocytes (PBL) as antigen-presenting cells (APC). There have been several reports that T cells can present antigen themselves. We have used tetanus antigen-specific T cell clones to examine the effects of envelope glycoproteins of HIV-1 on processing and presentation of antigen to T cells. Cloned T cells were pre-incubated with soluble crude preparation of tetanus antigen for 4 h at 37°C, irradiated, and used as APC (T-APC). These cells could present antigen, as assessed by the ability of the autologous cloned T cells to proliferate. Resting T cells and phytohaemagglutinin-activated T cell blasts from autologous PBL could not present tetanus antigen to the responder cloned T cells. Antigen presentation by T-APC was abrogated by treating cells with anti-HLA-DR but not by anti-HLA-DQ monoclonal antibodies; treatment of tetanus antigen-pulsed T-APC with anti-tetanus antibody also blocked the ability of these cells to induce proliferation in responder T cells. Antigen presentation by cloned T cells was by a chloroquine-resistant pathway. Pretreatment of T-APC with envelope glycoprotein of HIV-1, gp120. did not affect the proliferative responses of the responder T cells. These data suggest that gp 120 does not inhibit the antigen-presenting function while suppressing antigen-specific responses.
Degradation of aggregates of activated C3 (C3b) by monocytes of patients with rheumatoid arthritis is related to vasculitis
Authors: HEURKENS A H M , BREEDVELD F C , KEUR C VD , BRAND R , DAHA M R
Published: July 2010

We investigated whether a decreased complement receptor expression or function of monocytes isolated from peripheral blood of 52 patients with rheumatoid arthritis (RA) and rheumatoid vasculitis (RV) could account for the previously observed diminished degradation of immune complexes by monocytes of patients with RA and RV. On average, monocytes from all patients expressed significantly less CR1, and degraded significantly less AC3b when compared with monocytes of healthy controls. In addition, monocytes from RV patients degraded significantly less AC3b when compared with monocytes from patients with RA. The expression of both CR1 and CR3 on monocytes of RV patients was lower compared with RA patients but this difference was only significant for CR3. No differences were found between AC3b degradation and the expression of CR1 and CR3 between patients with active and inactive RA. Using linear discriminant analysis on the variables AC3b, CR1 and CR3, 94% of the patients could be classified correctly as healthy controls, RA or RV, suggesting a true multi-variate relationship between these parameters and patients groups. Our results suggest that the diminished capacity of monocytes from RA patients to degrade AC3b is due partly to a decreased expression of CR1 and CR3.
Immunological study of histologically non-involved jejunum during Crohn's disease: evidence for reduced in vivo secretion of secretory IgA
Authors: MARTEAU P , COLOMBEL J F , NEMETH J , VAERMAN J P , DIVE J C , RAMBAUD J C
Published: July 2010

We studied the local humoral immunity of histologically non-involved jejunum in patients with Crohn's disease. Normal subjects and patients with ulcerative colitis served as controls. Jejunal fluid samples were collected during segmental jejunal perfusion, under an occluding balloon and the in vivo jejunal secretion of the following proteins was determined: albumin, orosomucoid, transferrin, α2-macroglobulin, secretory component, monomeric and polymeric IgA, IgG, and IgM. The densities and number of IgA-, IgG-, and IgM-containing cells in the lamina propria of the jejunum were measured on perendoscopic biopsies. Jejunal secretion of polymeric IgA and the density of IgA-containing plasma cells in the lamina propria were significantly lower in patients with Crohn's disease than in both control groups. This abnormal intestinal immune response, which was not correlated to the activity of the disease, might be involved in its pathogenesis.
The specificity of the antibody response to internal antigens ofAscaris: heterogeneity in infected humans, and MHC (H-2) control of the repertoire in mice
Authors: KENNEDY M W , TOMLINSON L A , FRASER E M , CHRISTIE J F
Published: July 2010

Children from an area of Africa endemic for the large round worm of humans, Ascaris lumbricoides, were found to vary considerably in the specificity of their serum IgG response to the internal antigens of the parasite. This was particularly noticeable for responses to a 14-kD protein (ABA-I) of the parasite that has previously been shown to be the subject of a strong IgE antibody response in infected animals. The possibility that this heterogeneity in immune repertoire has a genetic basis was explored in inbred mice infected with Ascaris suum. This showed that no strain responded to all the potential antigens, that the recognition profiles of strains bearing independent haplotypes were unique, and only H-2-indentical strains had responses of similar specificities. Major histocompatability complex (MHC) restriction was confirmed using H-2-congenic animals on BALB and BIO backgrounds, which responded according to their H-2 haplotype. It is likely, therefore, that it is the MHC which controls the repertoire to Ascaris antigens in infected people. If this is so, then there will be implications for immunopathology associated with ascariasis. and possibly also for resistance and susceptibility to infection.
Tetrandrine, a plant alkaloid, inhibits the production of tumour necrosis factor-alpha (cachectin) hy human monocytes
Authors: FERRANTE A , SEOW W K , ROWAN-KELLY B , THONG Y H
Published: July 2010

Human mononuclcar leucocytes (MNL) or the adherent fraction (monocytes) produced tumour necrosis factor-alpha (TNF-α) (by EL1SA) in culture when stimulated with killed Staphylococcus aureus. The bisbenzylisoquinoline alkaloid, tetrandrine inhibited the capacity of MNL and monocytes to produce TNF-α at a concentration range of 0·1 to 5 μg/ml. Tetrandrine may be potentially useful in the treatment of inflammatory diseases in which TNF-α plays a major role.
Dual mechanism in induction of human neutrophil cytotoxicity: activation of protein kinase C and elevation in intracellular calcium
Authors: GAVIOLI R , SPISANI S , GIULIANI A L , TRANIELLO S
Published: July 2010

Human neutrophils can damage the non-immunized K562 cell line when stimulated by phorbol myristate acetate (PMA). The combination of the activation of protein kinase C by phorbol and the increase of free intracellular calcium by ionophore potentiates the lytic reaction. The OKT9-immunized target cells are not able to induce by themselves the lytic response in neutrophils, but by combining the two signals, the antigenic stimulus and PMA, a high level of cytolytic response is attained. The addition of EGTA does not affect neutrophil cytotoxicity against antibody-coated targets, while it markedly reduced the lytic reaction against non-immunized targets; in contrast, the addition of EGTA together with the ionophore ionomycin completely suppresses the lysis of immunized and non-immunized targets. The treatment of neutrophils with the protein kinase C inhibitor H-7 causes a dose-related inhibition of the lytic functions that is greater on unsensitized K562. Thus the interaction of Fc receptors with immunized targets is required for reaching the maximal cytolysis. The enhanced lytic activity that occurs in the presence of immunized targets is mediated by calcium flux, as detected by using the monoclonal antibody AB8.28 which binds to FcIII receptors (FcRIII), thus supporting that both signals are involved.
M5, a phosphoinositol-linked human myelomonocytic activation-associated antigen
Authors: GADD S J , MAJDIC O , KASINRERK W , STOCKINGER H , MAURER D , EHER R , KNAPP W
Published: July 2010

When investigating the previously described monoclonal antibody (MoAb) VIM-5, raised against THP1 cells and binding to human monocytes and granulocytes, we found that the antigen detected by this antibody, designated M5, becomes very strongly expressed on monocytes after overnight culture with phorbol myristate acetate (PMA) or lipopolysaccharide (LPS) but not with recombinant human interferon-gamma (rhIFN-γ). Granulocytes stimulated with formyl-methionyl-leucyl-phenylalinine (FMLP) become negative for binding VIM-5. Immature granulocytes from bone marrow do not express M5, thus its expression on granulocytes is differentiation linked. The antigen bound by VIM-S is sensitive to hydrolysis by phosphoinositol-specific phospholipase C (PI-PLC). The immunoprecipitated MS antigen on monocytes is a broad band, with a peak of 50 kD (unreduced) and two bands of S3 kD and 44 kD (reduced). We have therefore detected an antigen that is up-regulated on stimulated monocytes but, conversely, down-regulated on FMLP-stimulated granulocytes.
Failure of ADCC to predict HIV-associated disease progression or outcome in a haemophiliac cohort
Authors: DALGLEISH A , SINCLAIR A , STEEL M , BEATSON D , LUDLAM C , HABESHAW J
Published: July 2010

Antibody-dependent cell-mediated cytotoxicity (ADCC) has been described in a number of virus infections and occurs in HIV infection. In spite of numerous studies on the ability of HIV-positive sera to mediate ADCC, it remains unclear whether ADCC has any functional significance. Here we examine serial serum samples from a cohort of haemophilia patients who became infected from a common source in 1984, and show that there is no significant difference in ADCC values between those who remained asymptomatic and those who progressed to disease. This study does not compare effector cell function and clinical status and does not exclude activity against different target cell lineages in vivo.
Fine specificities of anti-nuclear antibodies in murine models of graft-versus-host disease
Authors: VAN DAM A P , MEILOF J F , VAN DEN BRINK H G , SMEENK R J T
Published: July 2010

Two models of murine graft-versus-host disease (GVHD) were studied with respect to autoantibody production and development of systemic lupus erythematosus (SLE) like disease. One model was induced by injection of (B10.A(4R) × B10.A(2R))F1 mice with parental (B10.A(4R)) spleen and lymph node cells (groups I GVHD), the other by injection of (DBA/2 × C57/B16)F1 mice with DBA/2 cells (group II GVHD). Group I GVHD mice remained in a seemingly healthy condition and did not show any proteinuria, in spite of high titres of anti-nuclear antibodies including antibodies to dsDNA, anti-Sm and anti-ribosomal P protein antibodies. Measured levels of these autoantibodies as well as their isotypes were comparable with those found in MRL/lpr and NZB/W mice. Group II GVHD mice developed SLE-like disease signs, including severe proteinuria. At 4 months after induction of the GVHD, almost 50% of these mice had died. At the time nephritis was present, group II mice also produced anti-dsDNA and anti-nuclear antibodies of other (unknown) specificities, but no anti-Sm or anti-P. Furthermore, the incidence of these antibodies was lower than observed in group I GVHD, MRL/lpr or NZB/W mice. It is concluded that (high avidity) anti-dsDNA as well as anti-Sm and anti-P may be present in the circulation without giving rise to the development of nephritis.
Recombinant 70-kD protein used for determination of autoantigenic epitopes recognized by anti-RNP sera
Authors: NYMAN U , LUNDBERG I , HEDFORS E , PETTERSSON I
Published: July 2010

A panel of high- and low-titre anti-RNP-positive patient sera was tested for reactivity with human snRNP proteins. The U1 snRNP-specific 70-kD peptide was found to be the most prominent RNP antigen recognized by high-titre anti-RNP sera, mainly found in patients with mixed connective tissue disease (MCTD). The reactivity with the 70-kD protein was further analysed with recombinant fusion proteins containing different segments of the protein. One major and two minor antigenic regions were found. Most patient sera, both with high- and low-titre anti-RNP antibodies, only recognized the major region. The most aminoterminal region, showing partial sequence homology with a mouse retroviral p30 gag protein, contained an epitope that was recognized by one serum only.
Autoantibodies to epithelial cells in patients on long-term therapy with leucocyte-derived interferon-alpha (IFN-α)
Authors: KARLSSON-PARRA A , BURMAN P , HAGBERG H , ÖBERG K , ALM G , KLARESKOG L , KARLSSON F A
Published: July 2010

During routine screening for anti-nuclear antibodies, using rat liver tissue as substrate, a reactivity against bile duct epithelium was observed in sera from carcinoid tumour patients given human leucocyte-derived IFN-a (HuLe IFN-a). In a retrospective study, initiated by this observation, the development of serum antibodies to bile duct epithelium was observed in nine out of 12 patients with carcinoid tumours and in three out of 14 patients with hairy-cell leukaemia during their treatment with HuLe IFN-α. However, no bile duct reactivity was observed in sera from carcinoid or hairy-cell leukaemia in patients given recombinant IFN-α. When analysing the reactivity of positive sera against a panel of rat and human tissues, a uniform reactivity was observed against simple epithelial cells lining the gastrointestinal tract, pancreatic secretory ducts, fallopian tube, kidney tubuli. mesothelium and also against carcinoid tumour cells. The mechanisms promoting autoreactivity against this simple epithelial cell auloantigen is so far unknown. The cytoplasmic as well as the restricted staining pattern of simple epithelial cells may indicate autoreactivity against certain cytoskelelal intermediate filaments, such as cytokeratin19, 18 and8, known to be exclusively present in simple epithelial cells and tumours derived from them.
Subpopulations of CD4+ cells in lpr/lpr mice: differences in expression of T cell receptor/CD3 complex and proliferative responses
Authors: ASANO T , YOSHIKAI Y , MATSUMOTO K , MATUZAKI G , NOMOTO K
Published: July 2010

CD4+ cells from autoimmune-prone C57BL/6 lpr/lpr mice contain two subpopulations, B220 CD4+ and B220+ CD4+ cells. Highly purified B220 CD4+ cells from C57BL/6 +/+ and lpr/lpr mice were examined by comparing functional characteristics and expression of cell surface antigens and T cell receptor (TcR)/CD3 complex. Both Ipr B220+CD4+ and B220+CD4− CD8 cells, most of which were PgP-1 positive, expressed TcR/CD3 complex on the cell surface at lower level as compared with B220 CD4+ cells of age-matched normal mice. In addition, the B220 CD4+ cells were heterogeneous on the basis of surface expression of PgP-1 and CD3 antigens. Normal levels of TcR Cx-, Cβ- and Vβ8-specific mRN A were found in the B220 CD4+cells and B220+ CD4+ cells as compared with normal B220− CD4+ cells, while Vβ8-specific mRN A was preferentially expressed only by B220+ CD4CD8 cells. Either B220+ CD4+ cells and B220+CD4− CD8− cells Tailed to respond to anti-CD3 monoclonal antibody (MoAb) as assessed by proliferative responses and production of interleukin-2 (IL-2). However, appreciable levels of reactivity to anti-CD3 MoAb were detected in the B220 CD4+ cells, although the responsiveness of this subset to such stimuli were reduced, compared with those of normal control. These results indicate that the B220− CD4+ cells in Ipr mice are phenotypically and functionally distinct from normal B220 CD4+ cells.
Sequential serological responses to Aspergillus fumigatus in patients with cystic fibrosis. Use of antigen ‘stretching’ to delineate IgG and IgE activity
Authors: EDWARDS J H , ALFAHAM M , FIFIELD R , PHILPOT C , CLEMENT M J , GOODCHILD M C
Published: July 2010

Immunogens from Aspergillus fumigatus were fractionated on the basis of molecular weight. Nine fractions ranging from 900 to 10 kD were used in ELISA and in a radioallergosorbent test (RAST) with sera from cases of allergic bronchopulmonary aspergillosis (ABPA) and from cystic fibrosis (CF) patients with ABPA or other Aspergillus involvement and compared with control subjects. The profile of IgG reactivity to the nine fractions did not vary substantially for all Aspergillus-involved groups producing peaks at > 900 kD and 170 kD whereas the profile for control subjects had a peak at > 900 kD only. The IgE profile for CF patients with ABPA did not differ from the profile of the RAST-positive CF patients without ABPA and provided only one peak of activity at 24 kD. Recovery from an episode of ABPA in CF patients was accompanied by a fall in both IgG and IgE antibody levels to all nine fractions, whereas increases in IgG and IgE to all fractions were seen during an episode of ABPA. Although there was an exaggerated IgG increase to antigens in the 43–170 kD range during ABPA, a meaningful increase was also observed to unfractionated A. fumigatus antigen preparations. With IgE in one detailed study the 24-kD fraction provided a better indication of Aspergillus involvement than the unfractionated A. fumigatus antigens. Sequential studies of IgG and IgE levels were not able to predict an episode of ABPA but were useful in conjunction with clinical assessment in following the course of the illness.
Passive transfer of immunity toMycobacterium aviumin susceptible and resistant strains of mice
Authors: STOKES R W , COLLINS F M
Published: July 2010

Naturally susceptible mice (C57BL/6) infected with M. avium (strain Weybridge) developed a population of splenic T cells which, on transfer to syngeneic recipient mice, conferred significant protection against a subsequent challenge inoculum of M. avium. Similar T cells from naturally resistant mice (A/J) did not protect syngeneic recipient mice. Growth of M. avium in donor mice only occurred in the C57BL/6 strain. Replication of M. avium in donor mice was necessary for the development of protective T cells. High numbers of killed mycobacterium did not induce immune T cells. In addition. A/J mice inoculated with increasing numbers of viable M. avium (which still did not replicate) failed to develop protective T lymphocytes. Further studies indicated that no protective T cells were present in the M. avium-infected A/J mouse, although evidence for non-specific immunity in these mice was obtained. In addition, BCG (which grows progressively in A/J mice) stimulated a population of splenic T cells which protected recipient mice from subsequent infection with M. tuberculosis.