bluuebunny/crossref_metadata_2025 huggingface dataset:
Transforming growth factor-β1 in rheumatoid synovial membrane and cartilage pannus junction
Authors: CHU C Q , FIELD M , ABNEY E , ZHENG R Q H , ALLARD S , FELDMANN M , MAINI R N
Published: July 2010

Transforming growth factor (TGF)-β has been shown to promote tissue repair and have immunosuppressivc actions, and has been proposed to have a role in rheumatoid arthritis (RA). Using immunohistochemical techniques with rabbit F(ab′)2 antibodies raised against recombinant human TGF-β1, we have detected TGF-β1 in the synovial tissue and cartilagc/pannus junction (CPJ) from 18/18 patients with RA. TGF-β1 was found predominantly in the thickened synovial lining layer in RA. but also detected in a peri vascular pattern in the synovial interstitium as well as in occasional cells in the lymphoid aggregates. At the CPJ it was found both in cells at the distinct junction as well as in the transitional region of the diffuse fibroblastic zone. The cells staining for TGF-β1 were identified by double immunofluorcsccnce staining as being from the monocy te/macrophage scries as well as the type B synovial lining cells. TGF-β1 was also detected in the synovial membrane sections from 4/4 patients with systemic lupus erythematosus/mixed connective tissue disease and 5/8 patients with osteoarthri-tis, in a similar distribution to that seen in RA, and in the lining layer of 1/7 normal synovial membranes. These results add to histological evidence confirming that TGF-/β1 is present in RA synovial cells and those from otherarthritides. The distributions of TGF-/β1 in RA synovial membrane reflects its known actions, as it can be detected at the CPJ, where it could induce repair, and close to activated cells upon which it may exert an immunosuppressivc action.
Extracellular organelles (prostasomes) are immunosuppressive components of human semen
Authors: KELLY R W , HOLLAND P , SKIBINSKI G , HARRISON C , MCMILLAN L , HARGREAVE T , JAMES K
Published: July 2010

Numerous reports have ascribed immunosuppressive activity to human seminal plasma and there is growing agreement that much of this activity can be accounted for by the very high levels of E series prostaglandins present (up to 300 μm 19-hydroxy prostaglandin E). However not all suppressive activity is due to prostaglandin since several reports have appeared of high molecular weight active substances and we have found that stripped seminal plasma is still effective in inhibiting the mitogen-induced proliferation of lymphocytes. In this study such immunosuppressive activity has been separated by molecular size fractionation and the activity has been found to be particulate and corresponded to the previously reported prostasomes. These are trilaminar to multilaminar vesicles (150 nm diameter) which are secreted by the prostate. Pure preparations of prostasomes inhibited mitogen-induced lymphoproliferation in a dose-dependent manner with a concentration of prostasomes equivalent to 40% of that seen in seminal fluid giving 69% suppression of thymidine incorporation. The suppressive activity survived boiling and therefore was unlikely to be due to enzymatic action associated with these organdies. Interaction with the accessory cells, involved in full development of the lymphoproliferation induced by mitogen, was indicated and this possibility was supported by the demonstration of a direct effect of prostasomes on macrophage function using a mouse macrophage cell line. The prostasomes in semen may play a complementary role to the prostaglandins in neutralizing the immune defences of the female reproductive tract. This combination would allow the alloantigenic spermatozoa the best chance of achieving fertilization, but at the same time leave the recipient open to any infection present in the semen.
Expression of rheumatoid factor associated cross-reactive idiotopes by glandular B cells in Sjögren's syndrome
Authors: DEACON E M , MATTHEWS J B , POTTS A J C , HAMBURGER J , MAGEED R A , JEFFERIS R
Published: July 2010

B cell expression of the germline gene-encoded, kappa IIIb-associated, rheumatoid factor (RF) cross-reactive idiotope (CRI) 17–109 and three VHI associated RF CRIs (G6, G8, H1) was investigated immunocytochemically in labial salivary glands from nine patients with primary and six with secondary Sjögren's syndrome, and in inflamed submandibular salivary glands from 10 patients with no history of connective tissue disease. Expression of CRIs by B cell infiltrates in labial glands from patients with primary and secondary Sjögren's syndrome were similar. Lymphoid infiltrates of labial glands from Sjögren's syndrome patients contained a higher proportion of kappa III+ cells reactive for the kappa IIIb-associated 17–109 idiotope (P<0.01) and larger G6 (P<0.02) and HI (P<0.01) positive B cell populations than those within inflamed submandibular salivary glands. Furthermore, in labial glands there was a significant correlation between numbers of 17–109 and G6 idiotope reactive cells (r = 0.61; P < 0.02), reflecting the known association between these H and L chain CRIs in RF IgM paraproteins. These results indicate that B cells bearing both VkIII and VHI-associated CRI are increased in the glandular infiltrates in Sjögren's syndrome and support the idea that this condition is associated with proliferation of immature B cell clones retaining germ-line V genes.
Inhibition of lymphocyte proliferation by a monoclonal antibody (RFT2) against CD7
Authors: COSTANTINIDES Y , KINGSLEY G , PITZALIS C , PANAYI G S
Published: July 2010

The effect of the monoclonal antibody RFT2, directed against CD7, on T cell proliferation in unseparated peripheral blood mononuclear cell populations induced by various stimulants was investigated. The addition of RFT2 to cell cultures inhibited the T cell proliferation induced by tuberculin PPD and OKT3 but not by phytohaemagglutinin, concanavalin. A or phorbol myristic acid; RFT2 had to be present during the first 24 h of culture in order to elicit inhibition; inhibition of proliferation was not due to down regulation of interleukin-2 receptor on the surface of T cells; and suppressive effects could be transferred by mononuclear cells prc-treated with RFT2- These results are of particular relevance in view of the known in rivo suppressive effect of RFT2 in humans.
Lymphokine-activated killer (LAK) cells modulate the effects of IL-2 on a T cell-mediated immune response
Authors: MCCULLOCH P , GALLAGHER G , WALSH L P , ZALOOM Y , XIE J
Published: July 2010

The ability of LAK cells and/or IL-2 to affect the course of an established T cell response was examined in a delayed-type hypersensitivity (DTH) model. IL-2 greatly increased the magnitude of the response at 24 h, while LAK cells alone had no effect. The administration of LAK cells and IL-2 together also had no effect on the magnitude of the DTH response, demonstrating that LAK cells were able to remove the enhancement seen with IL-2 alone. The presence of LAK cells reduced the serum half-life of IL-2 significantly, but not to an extent able to account for the observed loss of IL-2 induced DTH enhancement. IL-2 administration influenced cell phenotypes in the spleen and draining lymph nodes (DLN), as well as increasing splenic weight; the additional presence of LAK cells markedly altered these effects of IL-2 in the spleen (but not the DLN). Taken together, these results suggest that LAK cells interact with activated T-cells within the immune system and modulate their function.
Specific recognition pattern of IgM and IgG antibodies produced in the course of experimental paracoccidioidomycosis
Authors: VAZ C A C , MACKENZIE D W R , HEARN V M , CAMARGO Z P , SINGER-VERMES L M , BURGER E , CALICH V L G
Published: July 2010

Specific IgM and IgG responses to Paracoccidioides brasiliensis produced in resistant and susceptible mice during experimental paracoccidioidomycosis were examined by the immunoblotting procedure. Sera from infected mice recognized 51 antigen bands with apparent molecular masses From 8 to 86 kD. Sixteen of these were defined as major antigen bands because of almost universal presence of antibodies to them, and their intense staining. All sera, including those from normal control mice, tested for both IgM and IgG antibody reacted with the major E antigen which appeared as a large diffuse band from 43 to 47 kD. Comparisons between resistant and susceptible mice showed some significant differences in IgM responses to many antigen bands. While IgG responses were quite similar for both strains, differences were apparent in the response to the antigens at 62 and 68 kD.
Antineutrophil cytoplasmic antibodies (ANCA) directed against cathepsin G in ulcerative colitis, Crohn's disease and primary sclerosing cholangitis
Authors: HALBWACHS-MECARELLI L , NUSBAUM P , NOËL L H , REUMAUX D , ERLINGER S , GRÜNFELD J P , LESAVRE P
Published: July 2010

Autoantibodies directed against polymorphonuclear neutrophils (PMN) have been observed in serum from patients with ulcerative colitis (UC), Crohn's disease (CD) and primary sclerosing cholangitis (PSC) using indirect immunofluorescence and fixed granulocyte ELISA. Our study demonstrates the presence in the serum of these patients of autoantibodies which bind to an azurophilic granule component distinct from proteinase 3, elastase and myeloperoxidase. These autoantibodies thus belong to the ANCA family, but their antigen specificity differs from the already characterized ANCA antigens. We have found that the same ANCA antigen target, named UC-antigen, was recognized by serum IgG from patients with UC, CD and PSC. It was purified by Matrex Gel Orange A dye affinity chromatography and subsequent immunoabsorption of contaminant proteinase 3 with immobilized anti-proteinase 3 MoAb. The identity between this UC antigen and cathepsin G was demonstrated by their coelution from Matrex Gel Orange A column and the parallel titration of cathepsin G-specific enzymatic activity and UC ANCA binding, both in partially purified UC antigen and in highly pure cathepsin G. Furthermore, the use of cathepsin G ELISA confirmed that UC, CD and PSC patients' IgG did indeed bind to cathepsin G. Comparison of the results obtained with azurophilic granule- and cathepsin G-ELISA as well as inhibition of ANCA binding by anti-cathepsin G polyclonal antibodies, revealed that in some patients cathepsin G is the main azurophilic granule target of ANCA while others have other ANCA specificities. The fact that UC, CD and PSC are frequently associated with cathepsin G ANCA, while rarely occurring in other types of vasculitis, is intriguing but suggests that these diseases may have a common pathogenetic mechanism.
Antigen-specific T cell recognition of affinity-purified and recombinant thyroid peroxidase in autoimmune thyroid disease
Authors: EWINS D L , BARNETT P S , RATANACHAIYAVONG S , SHARROCK C , LANCHBURY J , McGREGOR A M , BANGA J P
Published: July 2010

TheTcell proliferative responses of peripheral blood lymphocytes from 20 patients with autoimmune thyroid disease (AITD) and 20 healthy controls were analysed to immunoaffinity-purified thyroid peroxidase (TPO) and recombinant antigen preparations generated in Escherichia coli as glutathione-s-transferase fusion proteins. The epitope specificity of the T cell response was investigated using a selection of eight discrete recombinant fragments encompassing the whole of the extracellular region of the TPO molecule. Significant differences in the proliferative responses between patients and controls were observed to the full length, affinity-purified TPO molecule (P < 0·002) as well as to the recombinant fragments Rlc (residues 145–250)(P<0·001) and R2b(residues 457–589)(P<0·001) suggesting the presence of at least two distinct T cell determinants on this autoantigen. One of these T cell epitopes, localized within the region Rlc, has not previously been identified by studies with synthetic peptides.
HLA-associated susceptibility to HIV-1 infection
Authors: FABIO G , SCORZA R , LAZZARIN A , MARCHINI M , ZARANTONELLO M , D'ARMINIO A , MARCHISIO P , PLEBANI A , LUZZATI R , COSTIGLIOLA P
Published: July 2010

We studied HLA antigen distribution of 50 heterosexual partners of HIV+ drug abusers with more than I year of sexual exposure to HIV, 36 children born to seropositive mothers and 61 haemophiliac patients exposed to presumably infectious clotting factor concentrates. B52 and B44 antigens were associated with HIV resistance while B51 was associated with HIV susceptibility. Forty-nine HIV+ drug abusers, spouses of heterosexual partners studied and 25 HIV+ mothers of the children were also typed. DR11 phenotype was associated with infectiousness of HIV+ subjects. Our data suggest that the HLA region controls susceptibility to infection with HIV and infectiousness of HIV+ subjects in different risk groups.
Abnormal production of B cell growth factor in patients with systemic lupus erythematosus
Authors: OGAWA N , ITOH M , GOTO Y
Published: July 2010

In order to clarify the role of B cell growth factor (BCGF) in the pathogenesis of systemic lupus erythematosus(SLE), BCGF production by peripheral blood mononuclear cells(PBMC) and T cells was studied using a new bioassay for BCGF activity. For this purpose, we established an Epstein-Barr (EB) virus-transformed B cell line KS-3.F10 that proliferates only in response to two B cell-specific BCGF, low-mol. wt BCGF (LMW-BCGF) and high-mol. wt BCGF (HMW-BCGF). PBMC from active SLE patients produced less BCGF when stimulated with phytohaemagglutinin (PHA) compared with controls. The decreased BCGF production by PHA-stimulated PBMC from active SLE reverted to control values when SLE became inactive. However, PHA-stimulated T cells from active SLE patients produced more BCGF compared with controls, whereas those from inactive SLE showed normal BCGF production. Spontaneous BCGF production by T cells was not observed in active SLE patients. These findings suggest that decreased BCGF production by SLE PBMC is due to excessive BCGF consumption by B cells in vitro and that SLE T cells produce large amounts of BCGF with appropriate immune stimuli in vitro to promote polyclonal B cell activation.
Cellular immune response to the cell walls ofMycobacterium lepraein leprosy patients and healthy subjects exposed to leprosy
Authors: ROCHE P W , NEUPANE K D , BRITTON W J
Published: July 2010

Cell walls of M. leprae consist of complex arrangements of carbohydrate, lipid, peptidoglycan and protein molecules. Recently, extractable proteins of a wide range of molecular weights were identified as components of the cell wall. We have examined the cellular immune responses of Nepali leprosy patients to a cell wall preparation of M. leprae enriched for these proteins. Strong lymphocyte proliferative responses to the antigens were present in half of the paucibacillary leprosy patients and in the majority of healthy control subjects with occupational exposure to leprosy. Patients with multi-bacillary disease responded poorly and patients with tuberculosis had intermediate responses. Proliferative responses to the cell wall protein fraction were strongly correlated to the proliferative responses to sonicates of the whole leprosy bacillus. Immunization of mice with cell wall proteins resulted in inhibition of growth of M. leprae following foot-pad inoculation with viable organisms. Therefore cell-mediated immune responses to the extractable proteins of the cell wall may play a role in protective immunity against M. leprae infection.
Identification of DNA-binding proteins on human umbilical vein endothelial cell plasma membrane
Authors: CHAN T M , FRAMPTON G , CAMERON J S
Published: July 2010

The binding of anti-DNA antibodies to the endothelial cell is mediated through DNA, which forms a bridge between the immunoglobulin and the plasma membrane. We have shown that 32P-labelled DNA bound to the plasma membrane of human umbilical vein endothelial cells (HUVEC) by a saturable process, which could be competitively inhibited by non-radiolabelled DNA. In addition, DNA-binding was enhanced in HUVEC that had been treated with IL-1α or tumour necrosis factor-alpha (TNF-α). DNA-binding proteins of mol. wt 46000, 92000, and 84000 were identified by the binding of 32P-labelled DNA to plasma membrane proteins separated on SDS PAGE. DNA-binding proteins of mol. wt 46000 and 84000 were also present in the cytosol and nucleus. Murinc anti-DNA MoAb 410 bound to a single band, at mol. wt 46000, of plasma membrane protein, in the presence of DNA. Our results showed that DNA-binding proteins are present in different cellular fractions of endothelial cells. DNA-binding proteins on the cell membrane could participate in the in situ formation of immune deposits: and their presence in the cell nucleus suggests a potential role in the modulation of cell function.
Regulation of endothelial adhesion molecules by ligands binding to the scavenger receptor
Authors: PALKAMA T , MAJURI M-L , MATTILA P , HURME M , RENKONEN R
Published: July 2010

Monocyte adherence to the endothelium, their penetration to the subendothelial space and excessive lipid accumulation (foam cell formation) are the initial events in atherogenesis. Scavenger receptors have been reported to play an important role in foam cell formation, since modified low density lipoproteins can be taken up via scavenger receptors in a non-down-regulated fashion. In this study we demonstrate that stimulation of scavenger receptors in endothelial cells induces the expression of endothelial adhesion molecules. Polyinosinic acid (poly I), a known scavenger receptor ligand, significantly induced the expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and E-selectin on human umbilical vein endothelial cells when compared with polycytidylic acid (poly C), a structurally related compound to poly I, which does not bind to the scavenger receptor. The effect of scavenger receptor ligands on the endothelial cell line EA hy. 926 was also tested. Poly I up-regutated ICAM-1 expression also on EA hy. 926 cells, whikit had no effect on IL-lβ or tumour necrosis factor-alpha (TNF-α) production on the same cell line. Poly I-induced ICAM-1 expression on EA hy. 926 cells could be inhibited by H7, a protein kinase C inhibitor, while HA 1004, a preferential protein kinase A inhibitor, had no effect on ICAM-1 expression. The role of protein kinase C in scavenger receptor-mediated adhesion molecule up-regulation was confirmed by the ability of poly I to directly activate protein kinase C, when measured with 3H-phorbol dibutyrate binding to EA hy. 926 cells, while poly C again was ineffective.
Rheumatoid factors from patients with rheumatoid arthritis react with Des-Lys58 -β2m, modified β2-microglobulin
Authors: WILLIAMS R C , NISSEN J R M H , MALONE C C
Published: July 2010

Ten polyclonal IgM rheumatoid factor (RF) preparations, affinity-purified from IgG columns, from patients with rheumatoid arthritis were studied for their ELISA reactivity with native β2m in parallel with Lys58-cleaved β2m and Des-Lys58-β2m, the latter representing cleavage products of the native molecule present in some pathologic human sera. Most RF showed positive reactions with the native form of β2m but reduced reactivity for the cleaved forms of β2m. Reactions between cleaved β2m and RF, in solution, were demonstrated by inhibition of RF binding to native β2m by prcincubation with a range of concentrations of Dcs-Lys58-β2m. By contrast, eight of nine murine MoAbs to human β2m showed approximately equivalent binding to native β2m, Lys58-cleaved β2m, and Des-Lys58-β2m. Reactions between individual human RF and the altered forms of β2m (Lys58-cleaved β2m and Des-Lys58-β2m) appeared to parallel the previously determined β2m single ammo acid specificities, in that RF’ showing strong reactivity with Lysine 58 also showed a significant diminished reactivity with the Des-Lys58-β2m lacking the critical lysine residue. The present studies demonstrate that while human RF react with Lys58-cleaved β2m or Des-Lys58-β2m, preferential reactivity is observed for native unaltered β2m.
Why do patients develop reactions to streptokinase?
Authors: LYNCH M , PENTECOST B L , LITTLER W A , STOCKLEY R A
Published: July 2010

Minor reactions to streptokinase are not uncommon, although the etiology is unknown. It is widely presumed, however, that these, like the more serious immune reactions, are antibody-mediated. We measured specific anti-streptokinase IgG. subclasses IgG1, IgG2, IgG3, IgG4 and IgE by ELISAs, haemagglutination, indirect Coombs' test and immunoblotting in six patients who developed reactions to streptokinase. Evidence of complement activation by streptokinase was sought by a haemolytic complement assay and by measurement of C3, C4 and C3d. The patients who reacted to streptokinase presented with low titres of anti-streptokinase IgG (median = 5; range 0–32) and IgG1 (median = 3; range 0–14). No evidence of any other IgG subclass was found, nor of specific anti-streptokinase IgE. Anti-streptokinase IgG1 was found to fix complement; patients who reacted to streptokinase were found to have low levels of total complement 1 year post reaction. Probable aggregates and fragments of human albumin (added stabiliser) were found in the streptokinase preparation and proved to be antigenic in some patients, but were not found to be related to the development of reactions. The findings suggest that patients who develop reactions to streptokinase cannot be predicted on the basis of antibody titres at presentation. Minor reactions to streptokinase would not appear to be antibody-mediated, although complement activation may be involved.
Common idiotypes expressed on human, monoclonal, abnormal immunoglobulins and cryoglobulins with polyreaetive autoantibody activities
Authors: BARBOUCHE M R , GUILBERT B , MAKNI S , GORGI Y , AYED K , AVRAMEAS S
Published: July 2010

Several hutiian monoclonal immunoglobtilins with the same auloatitibody activity have been shown to have cross-reactive idiotypes (CRI). In this study, using polyclonal anti-idiotypic antibodies, we fotind that 28% of hutnan monoclonal immunoglobulins with polyreaetive autoantibody activity from myeloma. Waldenstrom's macroglobulinaemia and eryoglobulinaemia patients shared common idiotype(s). Furthermore, the latter were expressed on human and murine natural MoAbs (respectively in 12% and 22% of the clones tested) and on human IgG preparations used for therapeutic intravenous injections (IVlg) and which contain natural antibodies. These findings suggest that monoclonal immunoglobulins could arise from the proliferation of a clotie that normally produces a natural antibody. The existenee of common idiotype(s) between monoclonal immunoglobulins and IVIg could be relevant to the improvement noted after treatment with IVIg in patients suffering from peripheral neuropathies associated with monoclonal gammopathy.
Elevated expressionin situof selectin and immunoglobulin superfamily type adhesion molecules in retroocular connective tissues from patients with Graves' ophthalmopathy
Authors: HEUFELDER A E , BAHN R S
Published: July 2010

Activation of certain adhesion molecules within vascular endothelium and the surrounding extravascular space is a critical event in the recruitment and targeting of an inflammatory response or autoimmune attack to a particular tissue site. We have recently demonstrated that the adhesion of lymphocytes to cultured retroocular fibroblasts obtained from patients with Graves’ ophthalmopathy (GO) is mediated predominantly by the interaction of lymphocyte function-associated antigen-1 (LFA-1), expressed on lymphocytes, with intercellular adhesion molecule-1 (ICAM-I), expressed by these cells following exposure to interferon-gamma (IFN-γ), tumour necrosis factor-alpha (TNF-α), IL-lα or purified thyroid-stimulating immunoglobulins. We now report the expression and localization in situ of several adhesion molecules. ICAM-1, endothelial leucocyte adhesion molccule-l (ELAM-1), vascular cell adhesion molecule-1 (VCAM-1), and LFA-3 in retroocular tissues derived from patients with severe GO (n=4) and normal individuals (n = 3). Serial cryostat sections of tissue specimens were processed for immunoperoxidase staining using various MoAbs against ICAM-1, ELAM-1, VCAM-1 and LFA-3. In addition, consecutive sections were stained with MoAbs against LFA-1, CD45RO (UCHL-1)DR-human leucocyte antigen (HLA-DR), CD11b/CD18 (Mac-1), and CD11c/CD18 (p150,95). In GO-retroocular tissues, strong immunoreactivity for ICAM-1 and LFA-3 was detected in blood vessels (>90%), in perimysial fibroblasts surrounding extraocular muscle fibres, and in connective tissue distinct from extraocular muscle. No ICAM-1 or LFA-3 immunoreactivity was present in extraocular muscle cells themselves. ICAM-1 and LFA-3 immunoreactivity in normal tissues was minimal or absent both in connective and muscle tissues. Vascular endothelium was strongly positive for ELAM-1 and VCAM-1 in GO-retroocular tissues, while VCAM-1 immunoreactivily was minimal (<5% of blood vessels) and ELAM-1 immunoreactivity was generally absent in normal retroocular tissue. LFA-1-expressing, activated mononuclear cells and memory T lymphocytes (CD3+/CD45RO+) were only detected in GO-retrocular tissues, and were mainly localized around blood vessels and in areas of ICAM-1-expressing connective and perimysial tissue. HLA-DR expression was restricted to GO-tissue specimens, with strong immunoreactivity detected in blood vessels, macrophages and connective tissue and perimysial fibroblasts. No HLA-DR was detectable in extraocular muscle cells. In conclusion, infiltration of the orbit in GO by mononuclear cells, and their targeting within the orbit, may depend upon the coordinate expression of certain adhesion and MHC molecules. Activation of appropriate adhesion molecules in mononuclear cells, endothelial cells and fibroblasts in response to various cytokines and inflammatory mediators may be involved in the evolution and perpetuance of GO.
Comparative analysis of biological activities ofDer p I-derived peptides on Fc receptor-bearing cells fromDermatophagoides pteronyssinus-sensitive patients
Authors: JEANNIN P , PESTEL J , BOSSUS M , LASSALLE P , TARTAR A , TONNEL A-B
Published: July 2010

The ability of Tour uncoupled synthetic peptides (p52–71, p117–133, p176–187, p188–199) derived from Der p I, a major allergen from the house dust mite Dermatophagoides pteronyssinus (Dpt) to stimulate FcεR+ cells from Dpt-sensitive patients was comparatively analysed. Each free peptide may specifically stimulate basophils (FcεRI+ cells) and platelets (FcεRII+ cells) from patients with significant levels of anti-Der p I IgE antibodies; p52–71 and p 117–133 appear the best cell stimulation inducers. Both concentration-dependent biological activities of Der p I-peptide on FcεR+ cells are enhanced by coupling peptide to a carrier (as human serum albumin). Interestingly each Der p I sensitive patient tested presents an individual pattern of response to peptide. Thus, from our results it appears that different Der p I sequences could be involved in the immune response to Der p I.
Antibodies to EYRKK vesicular stomatitis virus-related peptide account only for a minority of anti-Ro60kD antibodies
Authors: ROUTSIAS J G , SAKARELLOS-DAITSIOTTS M , DETSIKAS E , TZIOUFAS A G , SAKARELLOS C , MOUTSOPOULOS H M
Published: July 2010

Previous studies demonstrated a possible antigenic relation between the carboxyl terminal portion of anti-Ro60kD autoantigen and a nucleocapsid protein (N) of vesicular stomatitis virus (VSV). In order to investigate whether anti-Ro60kD autoantibodies react with the VSV homologous region of the Ro60kD protein we synthesized, according to Merrifield's method, the EYRKKMDI octapeptide (8p) sharing a common sequence with the N protein of VSV. Sera from 61 patients with autoimmune rheumatic diseases (34 systemic lupus erythematosus (SLE), 21 Sjögren's syndrome (SS) and six rheumatoid arthritis (RA)) as well as 59 from normal blood donors were tested for the presence of anti-Ro60kD autoantibodies by ELISA and immunoblot (IB) and anti 8p antibodies by ELISA. Antibodies to 8p were found in 9/31 of anti-Ro60kD IB-positive sera, 5/30 of anti-Ro60kD-negative sera and 2/59 of normal control sera. The concordance between the anti-8p ELISA and the anti-Ro60kD IB was very poor (χ2= 0·71 P= 0·4) in contrast to the anti-Ro60kD ELISA and the anti-Ro IB(χ2= 27·6, P= 10-7). Subsequent affinity purification of the anti-8p antibodies from a strong positive anti-8p and anti-Ro60kD SLE serum yielded 95% depletion of the anti-8p activity and 37% reduction of the anti-Ro60kD activity. Inhibition assays with the affinity-purified anti-8p antibodies demonstrated that the octapeptide gave 94·5% inhibition of the anti-Ro60kD activity, while Ro60kD protein led to 420% inhibition of the anti-8p. Preincubation of the serum with the octapeptide produced 4% inhibition of anti-Ro60kD ELISA. These results indicate that the anti-8p antibodies account only for a minority of the anti-Ro60kD autoantibodies.
MHC-associated immunopotentiation affects the embryo response to teratogens
Authors: Torchinsky A , Fein A , Carp HJA , Toder V
Published: July 2010

The present study was performed to evaluate whether the effect of environmental teratogens can be modified by maternal immunostimulation. Two chemicals, cyclophosphamide (CP) and 2,3-quinoxalinedimetanol,1,4-dioxide (QD) were used as the reference teratogens (RT), The response to these RT was investigated in two animal models: (i) primigravid C57B1 6 mice who underwent intrauterine immunization with allogeneic paternal (CBA/J), third-party (BALB c) or syngeneic male splenocytes 21 days before mating; (ii) C57B1 6 and CBA/J mice who were treated with RT during the second pregnancy only, after a different mating combination (syngeneic or allogeneic) in the first and the second pregnancy. Different doses of CP and QD were injected on days 12 and 9 of pregnancy, respectively. On day 19 of pregnancy implantation sites, resorptions, live and dead fetuses were recorded and live fetuses were examined for external and internal malformations with methods routinely used in teratological study. It was shown that intrauterine immunopotentiation with allogeneic paternal splenocytes clearly enhances the tolerance of F| embryos to RT, Thus, in CP-treated females the resorption rate and the proportion of malformed fetuses were significantly reduced. It was followed by an almost two-fold increase in fetal weight. The protective effect of such immunization in QD-treated females was manifested as a dramatic decrease of the proportion of malformed fetuses and the resorption rate, Syngeneic splenocytes could not significantly influence an embryo's sensitivity to RT, The response to RT was also significantly weaker in the second pregnancy of female mice mated twice allogeneically than that observed in allogeneically mated primigravid mice. These results show that the embryo's response to environmental teratogens may be influenced by fetomaternal immune interactions.
Intraglomerular expressions of IL-1α and platelet-derived growth factor (PDGF-B) mRNA in experimental immune complex-mediated glomerulonephritis
Authors: AKAI Y , IWANO M , KITAMURA Y , SHIIKI H , DOHI Y , DOHI K , MORIYAMA T , YONEMASU K
Published: July 2010

Both PDGF and IL-I play important roles as autocrine growth factors for cultured mesangial cells, and may be closely associated with the progression of glomerulonephritis. In this study we investigated intraglomerular expressions of PDGF-B and IL-1α mRNA in mice with bovine serum albumin (BSA) nephritis, a model of immune complex-mediated glomerulonephritis using the reverse transcription-polymerase chain reaction (RT-PCR) method. We also quantified intraglomerular PDGF-B mRNA by the competitive PCR and studied the correlation between the level of intraglomerular PDGF-B mRNA expression and the degree of observed glomerular injury. While expression of neither PDGF-B nor IL-lα mRNA was detected in glomeruli from control mice, both were strongly expressed in glomeruli from mice with BSA nephritis. IL-1α mRNA in glomeruli showed low accumulation in mice with mild glomerular injury, and was increased in mice with moderate glomerular injury. In contrast, high intraglomerular expression of PDGF-B mRNA occurred in all mice with mild glomerular injury and continued throughout the course of the disease. We observed no correlation between the level of PDGF-B mRNA expression and the histologic grade of renal damage. These results suggest that PDGF and IL-1 have different growth properties, and PDGF might play a role as a competence factor rather than a progression factor in the pathogenesis of immune complex-mediated glomerulonephritis.
Neonatal capsaicin pretreatment suppresses intramedullary inflammation in adjuvant-induced spondylitis
Authors: IMAI S , HUKUDA S , MAEDA T
Published: July 2010

In order to investigate the proposed involvement of neuropeptides in musculoskeletal inflammation we pretreated rats, in an adjuvant spondylitis model, with capsaicin, a neurotoxin. Immunohistochemistry showed that administration of capsaicin to newborn rats depicted irreversibly the neuropeptide substance P. Elimination of capsaicin-sensitive fibres by the neonatal injection of capsaicin did not suppress the peridiscitis of rats in which adjuvant spondylitis was induced at 7 weeks of age. However, elimination of capsaicin-sensitive fibres did suppress the inflammation usually seen in the bone marrow. We speculate that this intramedullary inflammation is normally induced or sustained by capsaicin-sensitive fibres.
Characterization of fresh (uncultured) tumour-infiltrating lymphocytes (TIL) and TIL-derived T cell lines from patients with renal cell carcinoma
Authors: MITROPOULOS D , KOOI S , RODRIGUEZ-VILLANUEVA J , PLATSOUCAS C D
Published: July 2010

Fresh (uncultured) TIL from 12 untreated patients with primary renal cell carcinoma were prepared from tumour specimens by enzymatic digestion, and were characterized by immunotluorescence using MoAbs recognizing leucocyte differentiation antigens or particular Va or Vα segments of the T cell receptor (TCR). These fresh TIL comprised CD3+ (20–84%); CD4+ (3–15%); CD8+ (13–35%); α/βTCR+ (20–50%); γδTCR+ (3–17%); CD16+ (1–18%) and CD56+ (3–10%) cells. Significant proportions of Vα2+, Vβ5.1+ and Vβ6+ cells were found in TIL of certain patients with renal cell carcinoma, suggesting that they comprised oligoclonal T cells. T cell lines were developed in low concentrations of rIL-2 (200 U/ml) from TIL from II patients with renal cell carcinoma, and were characterized by immunofluorescence and cell-mediated cytotoxicity. These T cell lines consisted primarily of CD3+ (51–94%); CD4+ (1–80%); CD8+ (0–84%); αβ/3TCR+ (65–87%); γδTCR+ (0–25%); CDI6+ (0–16%) and CD56+ (2–57%) cells. These Tcell lines exhibited non-specific cytotoxicity against autologous and aliogeneic renal tumour cells, with the exception of one T cell line that exhibited preferential cytotoxicity against autoiogous renal tumour cells. These results suggest that fresh TIL from patients with renal cell carcinoma contain significant proportions of oligoclonal T cells that may have accumulated at the tumour site as a result of a clonal expansion.
Hereditary C6 deficiency in a strain of PVG/c rats
Authors: LEENAERTS P L , STAD R K , HALL B M , VAN DAMME B J , VANRENTERGHEM Y , DAHA M R
Published: July 2010

A chance observation has led to the discovery of a strain of PVG rats (PVG/c−) which are deficient in complement (C) component C6. Analysis of total haemolytic activity (CH50) of PVG/c− serum revealed an absent CH50 activity compared with serum of other rat strains and of a PVG/c rat (PVG/c-) that showed normal C activity. Thus, the PVG/c− rat was unable to activate the C5b-9 membrane attack complex. To gain insight into the complement abnormalities, analysis of individual C components was performed. Testing the PVG/c− serum in a C6 haemolytic assay and using deficient human sera showed a deficiency of C6 in the PVG/c− rat. Highly purified human C6 and human sera deficient in other components were able to reconstitute the CH50 activity of the PVG/c+ rat. The possibility that an inactivator of C was present in PVG/c− serum was excluded. The deficiency was found to be inheritable and under the control of an autosomal recessive gene, Furthermore tissue antigens and immunity of the PVG/c+ rat were found to be identical to those determined in the PVG/c+ rat. With regard to their health status, the PVG/c− animals seem to have no disadvantages compared with PVG/c+ rats when held under the same conditions within the protected environment of animal facilities. Taken together, both rat strains provide an unique animal model for studying the biological role of C, particularly the C5b-9 membrane attack complex in experimental medicine.
Glycation of monoclonal antibodies impairs their ability to bind antigen
Authors: KENNEDY D M , SKILLBN A W , SELF C H
Published: July 2010

As elevated levels of glycated IgG have been detected in the plasma of patients with diabetes mellitus, a disease associated with increased susceptibility to infection, we have investigated whether glycation of MoAbs affects the kinetics and/or affinity of antigen binding. Three mouse MoAbs were incubated with 0.5 M glucose at pH 7.4 for 14-1 days at 37°C. Control MoAbs were incubated using identical conditions but with no added glucose. Using a surface plasmon resonance technique we found that glycation significantly increased the rate of dissociation (kdiss) of the antigen-antibody complex for all three MoAbs (P < 0.05, n= 4), but had no significant effect on the rate of association (kass). For one of the MoAbs, against human IgG (Fab), we also measured kdiss by an alternative method utilizing radiolabelled antigen, which confirmed that glycation of the antibody significantly increases kdiss (P < 0.001. n = 8). We also found using an ELISA-based method that glycation of the same MoAb significantly increased (he equilibrium dissociation constant (kd) (P < 0.05, n = 6). A significant increase in kd was observed after glycation using glucose concentrations consistent with those found in poorly controlled diabetics (P < 0.02, n = 5). We conclude that in vitro glycation can significantly lower the affinity of an antibody for its antigen, and significantly increases the rate of dissociation of the antigen antibody complex.
Elevated tumour necrosis factor-alpha (TNF-α) biological activity in psoriatic skin lesions
Authors: ETTEHADI P , GREAVES M W , WALLACH D , ADERKA D , CAMP R D R
Published: July 2010

Lesions of the common inflammatory skin disease psoriasis are characterized by epidermal hyperproliferation, leucocyte adhesion molecule expression and leucocyte infiltration. The local release of proinflammatory cytokines. such as TNF-α, may play an important role in the induction of these events. We have, therefore, analysed aqueous extracts of lesional and uninvolved (clinically normal) stratum corneum for the presence of TNF-α immunoreactivity and biological activity. TNF-α immunoreactivity and bioactivity were consistently higher in lesional compared with uninvolved samples. By using an anti-TNF-α neutralizing antibody it was demonstrated that the biological activity measured was due to the presence of TNF-α alone. Concentrations of soluble TNF receptors (p55 and p75) were also higher in lesional stratum corneum extracts, with the p55 form predominating. The plasma of psoriatic patients was also found to contain elevated concentrations of soluble p55 compared with normal controls. These results confirm the presence of immunoreactive TNF-α and, for the first time, conclusively demonstrate TNF-α biological activity and quantifiable concentrations of soluble TNF receptors (p55 and p75) in lesional psorialic samples. TNF-α recovery from stratum corneum probably reflects synthesis in deeper, viable layers, where il is likely lo exert its biological effects. Local and systemic release of soluble TNF receplors, in particular p55, may serve to regulate the effects of TNF-α in psoriasis.
Apparent killing of Mycobacterium tuberculosis by cytokine-activated human monocytes can be an artefact of a cytotoxic effect on the monocytes
Authors: WARWICK-DAVIES J , DHILLON J , O'BRIEN L , ANDREW P W , LOWRIE D B
Published: July 2010

A protocol using combined exposure to interferon-gamma (IFN-γ), calcitriol and tumour necrosis factor-alpha (TNF-α) has been reported to activate human monocytes in vitro to kill Mycobacterium tuberculosis. We have attempted to repeat the findings in two laboratories, with negative results: treated cells were no different from untreated cells in this respect. However, the treated cells were more sensitive to a toxic effect of the bacteria. We suggest that the reported dramatic mycobacterial killing may have been an illusory consequence of the toxicity leading to cell lysis and loss of the liberated bacteria from the assay.
Local synthesis of both macrophage and T cell cytokines by synovial fluid cells from children with juvenile rheumatoid arthritis
Authors: EBERHARD B A , LAXER R M , ANDERSSON U , SILVERMAN E D
Published: July 2010

The production of tumour necrosis factor-alpha (TNF-α), TNF-β and IL-6 in synovial fluid was studied in 50 samples of synovial fluid from 44 children with juvenile rheumatoid arthritis (JRA) by identifying cytokine production al a single-cell level. Post Ficoll-separated synovial fluid mono-nuclear cells were permeabilized and then intracellular TNF-α, TNF-β and IL-6 protein production was examined using indirect immunofluorescence and murine anti-cytokine MoAbs. All three cytokines were measured in 37 of the 50 samples. In 25 of the 37 samples there was complete concordance: all three cytokines were present in six and absent in 19 samples. At least one cytokine was present in 27/50 (54%) of synovial fluid samples. Overall, TNF-α was detected in 22/49 (45%) samples. TNF-β in 15/41 (37%) and IL-6 in 16/45 (36%) samples. Five patients had serial arthrocentesis, and in these samples there were two patients who had initially positive cytokine production, which on subsequent measurement was negative; in the other three patients there was no change from the previous cytokine production. We provide evidence that synovial fluid mononuclear cells produce monocyte and T cell cytokines in JRA. These findings suggest a role for both T cell and macrophage products in the pathogenesis of JRA, and the potential for modulation of cytokine production as a target for therapeutic intervention.
Binding of proteinase 3 and myeloperoxidase to endothelial cells: ANCA-mediated endothelial damage through ADCC?
Authors: BALLIEUX B E P B , ZONDERVAN K T , KIEVIT P , HAGEN E C , VAN ES L A , VAN DER WOUDE F J , DAHA M R
Published: July 2010

Binding of both proteinase 3 (PR3) and myeloperoxidase (MPO) to endothelial cells (EC) has been suggested to be involved in the vascular damage seen in patients with Wegener's granulomatosis or microscopic polyangiitis. In the present study we investigated in detail the interaction of MPO and PR3 with cultured human umbilical vein endothelial cells (HUVEC) and its matrix products. In addition, we investigated whether interaction of PR3 or MPO with HUVEC monolayers also resulted in antibody-dependent cell-mediated cytotoxicity (ADCC) mediated by anti-neutrophil cytoplasmic antibody (ANCA)-positive patient sera or rabbit IgG anti-PR3 or anti-MPO. Preincubation of HUVEC monolayers with PR3 or MPO resulted in a dose-dependent binding of both PR3 and MPO. However, HUVEC, preincubated with PR3 or MPO, followed by ANCA or by rabbit anti-PR3 or anti-MPO, were not susceptible to ADCC, Detailed analysis of the binding of PR3 to HUVEC monolayers showed that PR3 binds primarily to the extracellular matrix of endothelial cells, and to a very limited extent to the cells themselves. For MPO it was shown that it binds both to the extracellular matrix and to the endothelial cells themselves. However, after binding to HUVEC cultures, MPO was not detectable by polyclonal rabbit or human antibodies specific for MPO, probably because MPO is bound to sites not accessible for immunoglobulins. Binding of PR3 to HUVEC cultures (cells + matrix) was inhibited by fetal calf serum and by α1,-antitrypsin, but inactivation of enzymatic activity of PR3 by PMSF did not influence binding of PR3 to HUVEC cultures. Binding of MPO to HUVEC cultures was not influenced by fetal calf serum.
Effect of cytokine-induced soluble ICAM-1 from human synovial cells on synovial cell-lymphocyte adhesion
Authors: SHINGU M , HASHIMOTO M , EZAKI I , NOBUNAGA M
Published: July 2010

The present study was designed to establish (i) the effects of cytokines on soluble ICAM-1 (sICAM-1) production by human synovial cells (SC) and ICAM-1 expression on these cells, and (ii) the effects of sICAM-1 on lymphocyte-SC adhesion. sICAM-1 production was enhanced in parallel with ICAM-1 expression by IL-1β, TNF-α and IFN-γ. IL-4 showed no effects on ICAM-1 expression. In contrast with the transient elevation of cell-associated ICAM-1 by IL-1β, which peaked 36 h after stimulation and declined thereafter, sICAM-1 continued to accumulate in culture supernatants even after 48 h. Purified sICAM-1 was obtained from a 48h culture synovial cell supernatant by affinity chromatography using ICAM-1 monoclonal antibody. The purified sICAM-1 significantly inhibited adhesion of lymphocytes and monocytes to cytokine-stimulated synovial cells. These results suggest that sICAM-1 may modulate chronic synovitis by inhibiting ICAM-1-mediated cell-to-cell adhesion.
Diverse T cell receptor β gene usage by infiltrating T cells in the lacrimal glands of Sjögren's syndrome
Authors: MIZUSHIMA N , KOHSAKA H , TSUBOTA K , SAITO I , MIYASAKA N
Published: July 2010

Sjögren's syndrome (SS) is an autoimmune disease characterized by T cell infiltration into the salivary and lacrimal glands (LG). Previous studies on T cell receptor (TCR) usage in the minor salivary glands (SG) have yielded controversial results. We studied TCR β gene usage of the T cells infiltrating to LG, which is the other major target organ of SS. Total RNA was extracted from fresh LG and SG biopsy samples, and peripheral blood mononuclear cells from five SS patients, and converted to cDNA. TCR Vβ gene repertoire was then assessed with quantitative polymerase chain reaction (PCR) assay. Oligoclonality was studied by sequencing V-D-J junctional regions of the PCR products. The TCR Vβ gene usage in LG was diverse in every patient irrespective of disease duration, and similar to that of peripheral lymphocytes from a corresponding patient. The junctional region sequences of over-expressed Vβ families in LG T cells were heterogeneous. We did not find any identical clones shared by LG, SG and peripheral blood. These results showed that the infiltrating T cells in LG of SS patients are polyclonal, and LG and SG do not share the same dominant T cell clonotypes. These suggest that TCR-targeled disease manipulation may have a limited effect on SS.