bluuebunny/crossref_metadata_2025 huggingface dataset:
Early accumulation of suppressor cell precursors in the spleen ofMycobacterium lepraemurium-infected mice and analysis of theirin vitro-induced maturation
Authors: LEMIEUX S , GOSSELIN D , LUSIGNAN Y , TURCOTTE R
Published: July 2010

Spleen cells harvested from mice infected intraperitoneally with M. lepraemurium 11–17 weeks prior to harvest acquired the capacity to inhibit concanavalin A (Con A) induced proliferation of normal spleen cells when precultured for up to 24 h in mitogen-free medium. The in vitro induced suppressor activity correlated with the length of the preculture period, the time post-infection and the infecting dose. These findings were interpreted as an indication that suppressor cell precursors accumulated in the spleen of infected mice during the early phase of the disease. The interaction of infection-dependent adherent suppressor cell precursors and infection-independent, non-adherent regulatory cells is necessary for the suppressor activity to develop. Both the cells which transmit the inductive signal and the precursor cells which mature into active suppressor cells are radiosensitive, whereas suppressor activity itself is a function of radioresistant adherent cells. Preculture of cells for a short period, before they were cocultured with Con A-stimulated normal spleen cells, allowed the detection of suppressor cells before they were deleterious to the infected host and also turned out to be a relevant in vitro model for characterization of suppressor cell development during M. lepraemurium infection.
Quantitative measurement of SC5b-9 and C5b-9(m) in infarcted areas of human myocardium
Authors: HUGO F , HAMDOCH T , MATHEY D , SCHÄFER H , BHAKDI S
Published: July 2010

Previous immunohistochemical work has indicated that terminal C5b-9 complement complexes are selectively deposited in infarcted areas of human myocardium. In the present study, we sought to quantify C5b-9 levels in myocardial tissue, and to differentiate between the membrane-bound C5b-9 (m) and the cytolytically inactive SC5b-9 complex. Paired tissue specimens from infarcted and non-infarcted myocardium were obtained from 36 autopsies. The homogenized and washed tissues were extracted with n-octyl-β-D-glucopyranoside (octylglucoside) detergent, and the concentrations of C5b-9 in the extracts were determined by ELISA. Membrane-derived C5b-9 (m) and SC5b-9 were differentiated from each other on the basis of their characteristic sedimentation behaviour in sucrose density gradients. It was found that infarcted myocardial tissue contained on average an approximately three-fold higher concentration of C5b-9, compared with non-infarcted tissue. This increase was due in part to an increase in levels of C5b-9 (m). The results corroborate previous immunohistochemical data and show that complement activation occurs to completion with the generation of potentially cytotoxic C5b-9 complexes in infarcted myocardial tissues.
Selective IgG subclass deficiency: quantification and clinical relevance
Authors: JEFFERIS R , KUMARARATNE D S
Published: July 2010

Each of the four human IgG subclasses exhibits a unique profile of effector functions relevant to the clearance and elimination of infecting microorganisms. The quantitative response within each IgG subclass varies with the nature of the antigen, its route of entry and, presumably, the form in which it is presented to the immune system. This results in antibody responses to certain antigens being predominantly or exclusively of a single IgG subclass. An inability to produce antibody of the optimally protective isotype can result in a selective immunodeficiency state. This is particularly apparent for responses to certain bacterial carbohydrate antigens that are normally of IgG2 isotype. A failure to produce the appropriate specific antibody response may result in recurrent upper and/or lower respiratory tract infection. Careful patient investigation can identify such deficiencies and suggest appropriate clinical management. In this review we outline the biology and clinical relevance of the IgG subclasses and summarize current rational treatment approaches.
Longitudinal study of islet cell antibodies and insulin autoantibodies and development of diabetes in non-obese diabetic (NOD) mice
Authors: REDDY S , BIBBY N , ELLIOTT R B
Published: July 2010

We have previously shown the presence of circulating islet cell cytoplasmic antibodies (ICA) and insulin autoantibodies (IAA) in the NOD mouse before onset of insulin-dependent diabetes mellitus (IDDM). Here we have determined the levels of the two autoantibodies in 28 female NOD mice longitudinally from approximately day 40 to day 250, to examine their ontogeny, association and predictive value for diabetes. All animals (11 diabetic, 17 non-diabetic) showed varying levels of ICA at some stage, while IAA activity was found in 21 out of 28 mice. Expression of both the markers was seen in more than half of the animals by day 60, with higher levels and rates occurring subsequently in both diabetic and non-diabetic groups. The expression of ICA did not always correlate with that of IAA. There was no apparent difference in the ontogeny of ICA and IAA between the two groups. During the study period the number of animals with ICA was similar in the two groups, while the number of those with IAA was higher in the diabetic animals. In this group declining and rising levels of ICA were seen just before clinical diabetes with frequent peaks of IAA. In the same animals, eight out of 11 mice showed co-expression of high levels of both markers either intermittently or persistently prior to onset, whereas only one non-diabetic animal showed this. We conclude that the ontogeny and serum level of ICA or IAA alone could not be used to predict the clinical onset of diabetes in these animals. However, co-expression of high levels of both markers prior to onset may suggest a strong predisposition to clinical diabetes. This may have relevance to attempts to predict the onset of IDDM in humans who have one or both of these immunological markers.
Therapeutic effect of 15-deoxyspergualin on the progression of lupus nephritis in MRL mice. I. Immunopathological analyses
Authors: ITO S , UENO M , ARAKAWA M , SAITO T , AOYAGI T , FUJIWARA M
Published: July 2010

The effect of 15-deoxyspergualin (DSP), a newly developed immunosuppressive agent, on the development of spontaneously occurring lupus glomerulonephritis in MRL-lpr mice was examined. Administration of the drug was initiated at the age of 13 or 17 weeks, when polyclonal B cell activation and lupus nephropathy were apparent or became prominent. Treatment with DSP for up to 19 weeks of age at a dose of 2 mg/kg twice a day or 5 mg/kg daily strongly suppressed the increment of IgG-producing cell numbers in the spleen and serum levels of immune complexes and anti-DNA antibodies. Glomerular histoiogical score estimated by light microscopy and IgG and C3 deposition in renal glomeruli were improved, compared with untreated control mice. Thus, DSP was shown to suppress the progression of polyclonal B cell activation and lupus nephropathy in MRL/lpr mice. These results suggest that DSP may be used as a therapeutic agent for systemic lupus erythematosus.
Vβ5 andVβ8 memory T cells in adults and infancy: co-ordinated increase in response to early antigen stimulus
Authors: HAYWARD A R , CLARKE J , COSYNS M
Published: July 2010

Blood mononuclear cells (MNC) expressing the T cell receptor Vβ gene families were identified with the monoclonal antibodies IC1 (Vβ5) or Mx6 (Vβ8). These cells were typed for expression of the T200 common leucocyte antigen with the CD4SRA antibody defining the 220-kD variant (which characterizes naive cells) and theUCHL1 antibody which defines the 180-kD variant (characterizing memory cells). Of adult MNC, 3±0.44% stained for Vβ5 and 4.12±0.26% stained for Vβ8; similar results were obtained in a group of infants aged less than 3 months, who were exposed to antigen stimulus in the form of infection and/or blood transfusion. While the total percentages of Vβ5 and Vβ8 cells in the blood of adults were not correlated, the proportions of these cells expressing CD4SRO was positively correlated. A similar trend which did not reach statistical significance was present in a group of infants. A minority of the infants studied showed transient deviations from the mean established for adults. The results suggest that naive and memory T cell populations have similar frequencies of Vβ5 and Vβ8 expressing cells, and that age does not affect the frequency of usage of either of these Vβ families.
Monoclonal antibody BA-1 to the human B lymphocyte marker CD24 recognizes a sialic acid (N-acetylneuraminic acid) dependent epitope in multi-valent display on peptide
Authors: MEHMET H , LARKIN M , TANG P W , LEBJEN T W , FEIZI T
Published: July 2010

Evidence is presented that monoclonal antibody BA-1, directed against a marker (CD24) of human lymphocytes of B cell lineage, recognizes a sialic aeul-dependent cpitopc. This conclusion is based on a scries of experiments exploiting the reaction of this antibody with bovine and ovine submaxillary mucins. Expression of the cpitopc was enhanced following alkaline saponification of bovine submaxillary mucin, which converts 0-acctylatcd ncuraminic acid residues to N-acelylncuraminic acid. The epitope was destroyed following ncuraminidase or mild acid treatment of the mucins, and its expression was diminished following neuraminidasc treatment of B lymphobtastoid cells. Glycopcptidcs obtained by digestion of the bovine mucin with papain, trypsin or pronasc were lacking in antigenicity. However, anligcnic activity could be regenerated after conjugation of pronase glycopepiides to poly-L-lysinc. These results indicate that multivalenl display of sialo-oligosacchar-ide on peptide rather than a protease-susceptible polypcplide domain is required for BA-1 antibody binding.
Adhesion of polymorphonuclear cells to human endothelial cells. Adhesion-molecule-dependent, and Fc receptor-mediated adhesion-molecule-independent mechanisms
Authors: LEEUWENBERG J F M , JEUNHOMME G M A A , BUURMAN W A
Published: July 2010

Activation of human umbilical vein endothelial (HUVE) cells with the inflammatory mediators tumour necrosis factor-alpha (TNF), interleukin-1 (IL-1), lipopolysaccharide (LPS) and phorbol esters enhanced their adhesiveness for leucocytes. The appearance of an activation antigen ELAM-1, recognized by a monoclonal antibody (MoAb) ENAI, parallels the kinetics of the enhanced adherence of leucocytes to endothelial cells. Adhesion of polymorphonuclear cells (PMN) to activated HUVE cells could be blocked by F(ab)2 fragments of MoAb ENA1 up to 60%. An additive inhibition of the adhesion was established by pre-incubation of the PMN with anti-CD18 MoAb and J. or leucocyte adhesion inhibitor (LAI), produced by endothelial cells. An opposite reaction, however, was observed when HUVE cells were pre-incubated with intact MoAb ENAI, resulting in an enhancement of the adhesion up to 200%. Apparently, the blocking effect of MoAb ENA1 could be bypassed by the strong interaction of the Fc part of the MoAb with the Fc receptor (FcR) on the PMN. Similarly, anti-CD 18 MoAb and J. or LAI reduced the adhesion observed if intact ENAI were used, and Fc-FcR interaction took place. The results presented in this study indicate that adhesion via ELAM-1, the CD18 antigen and via the receptor for LAI are different mechanisms. These mechanisms may act in concert to strengthen the binding of PMN to HUVE cells. Moreover, a strong adhesion could be established via the Fc part of MoAbs directed against HUVE cells with the FcR on the PMN. The phenomenon described may play a roie in graft rejection and in diseases where antibodies directed against endothelium are involved.
Monoclonal autoantibodies from insulin-dependent diabetic patients: autoantibodies against β-cell surface or cytoplasmic antigens
Authors: HOUSSAINT E , POGU S , OUARY M , SAI P
Published: July 2010

To investigate the target antigen(s) recognized during the autoimmune process in insulin-dependent diabetes mellitus (IDDM), we produced human monoclonal antibodies by Epstein-Barr virus transformation of peripheral blood lymphocytes from a large number (n= 50) of newly diagnosed IDDM patients. Screening by indirect immunofluorescence assay, using the RINm5F rat insulinoma cell line and eight other human or rat tumour cell lines, was performed to identify monoclonal antibodies that reacted with either membrane or cytoplasmic antigens. Eighteen IgM monoclonal antibodies reacting with cytoplasmic antigens of RIN cells were obtained; 14 of them also showed a staining of the cytoplasm of various non-β-cell lines, while four displayed a binding restricted to β-txlls among the panel tested. However, among three monoclonal antibodies reacting with the membrane of RIN cells, one (HMD-1) produced an IgG antibody with a binding restricted to the membrane of β-cells (RIN, HIT, and normal rat islet cells). The membrane antigens of HMD-1 were identified in Western blotting as proteins with molecular weights of 64 and 70 kD. This antibody had no apparent cytotoxic effect on RIN cells. These data suggest that, apart from ‘natural autoantibodies,’ it is feasible to obtain human monoclonal antibodies from IDDM patients that bind specifically to the β-cell cytoplasm or to the β-cell membrane.
Autoantibodies to crude human leucocyte interferon (IFN), native human IFN, recombinant human IFN-alpha 2b and human IFN-gamma in healthy blood donors
Authors: ROSS C , HANSEN M B , SCHYBERG T , BERG K
Published: July 2010

Recently, naturally occurring antibodies to IFN-α were discovered in a few systemic lupus erythematosus (SLE) and cancer patients; however, in most patients monitored for anti-IFN antibodies before treatment, no antibodies were found. In an attempt to explain the ‘IFN-blocking effect’ that we observed in all serum samples we investigated 200 sera from healthy blood donors. We isolated the globulin fraction, and used rabbit anti-human IgG and IgM columns, protein A columns and T-gel affinity chromatography to isolate human IgG and IgM. All sample fractions were tested in a biological IFN neutralization assay by means of a sensitive MTT-assay. We found that normal human serum contained autoantibodies to crude human leucocyte IFN, native human fibroblast IFN, recombinant human leucocyte IFN-α2b and recombinant human IFN-γ, and that these naturally occurring antibodies were biologically active immunoglobulins of IgG and IgM type. These anti-IFN antibodies were also present in purified human normal immunoglobulin pools. We conclude that all humans have naturally occurring anti-interferon antibodies in their serum, and it is a tempting theory that human cytokines and lymphokines are, at least partly, regulated by immunoglobulins.
Thyroid follicular cell function after non-lethal complement membrane attack
Authors: WEETMAN A P , FREEMAN M , MORGAN B P
Published: July 2010

Terminal complement complexes have been identified around thyroid follicles in Graves' disease and Hashimoto's thyroiditis. and the concentrations of such complexes are increased in the sera of these patients, suggesting a role for complement activation and membrane attack complexes (MAC) in autoimmune thyroiditis. This has been investigated further using cultured human and rat thyroid cells. Thyrocytes were resistant to lysis by homologous complement, in contrast to the effects of heterologous (rabbit) complement. The formation of non-lethal amounts of MAC, using reactive lysis or classical pathway activation, significantly reduced cAMP production by these cells in response to thyroid-stimulating hormone (TSH) (P<0.01); similar effects were seen with thyroid-stimulating antibodies. Thyroid cells were able to recover rapidly from complement attack after washing and incubation for 30 min. Non-lethal MAC formation also resulted in reactive oxygen metabolite production, detected by luminol-dependent chemiluminescence in three out of five thyroid cell preparations tested, Ionomycin, but not TSH. also stimulated reactive oxygen metabolite production. These results suggest that repeated or continuous sub-lethal complement attack on thyroid cells may exacerbate hypothyroidism in Hashimoto's thyroidilis, or partially counter the effects of thyroid-stimulating antibodies in Graves' disease. Furthermore, the production of reactive oxygen metabolites in these circumstances could increase the intra-thyroidal inflammatory response; oxygen radical scavenging by anti-thyroid drugs (which are concentrated by thyrocytes) may account in part for the amelioration of thyroidilis observed with such treatment.
Degradation of human myelin in vitro by leucocytes from patients with multiple sclerosis
Authors: OWEN S J , WATSON C M , DAVISON A N
Published: July 2010

In order to study the possible autoimmune basis of multiple sclerosis (MS) a quantitative method has been used to investigate breakdown of human myelin in vitro. We found that serum from MS patients and controls was generally devoid of any myelin degradative activity. However, isolated peripheral blood mononuclear cells from 43% of MS patients showed significant myelin degradative activity as did those from 61.5% of patients with rheumatoid arthritis (RA). Myelin degradation by cells was found in only 13% of patients with other neurological diseases and in no healthy controls. It is proposed that this non-specific peripheral cellular immune degradative activity originates from cells activated within the central nervous system of MS patients or the joints of individuals with RA. As a result, activity in the blood only indirectly reflects the ongoing inflammatory response at the primary site, accounting for the lack of correlation between changes in the blood and the clinical status of the MS patient. We further propose that the lack of in vitro myelin degradative activity in cells recovered from the cerebrospinal fluid is due to autoaggressive cells being sequestered to the brain.
Isolation and characterization of a high molecular weight lymphocyte Feγ receptor-blocking factor associated with renal allograft survival
Authors: SANDILANDS G P , COCKER J E , McMILLAN M A , OWSIANKA A M , MARSDEN H , JUNOR B J R , BRIGGS J D , MACSWEEN R N M
Published: July 2010

We have confirmed our previous observation that improved human renal allograft survival is associated with the presence in pre-transplant serum of a high molecular weight lymphocyte Fey receptor-blocking factor. Serum fractionation studies suggest that this factor is a complex protein consisting of IgG together with an IgG-binding protein which has an apparent molecular weight of approximately 60 kD.
In vivoboosting of lung natural killer and lymphokine-activated killer cell activity by interleukin-2: comparison of systemic, intrapleural and inhalation routes
Authors: FLEXMAN J P , MANNING L S , ROBINSON B W S
Published: July 2010

Natural killer (NK) cells are thought to play a role in host defence against malignancy and infection, in immunoregulation and as precursor cells in a generation of lymphokine-activated killer (LAK) cells which can lyse NK-resistant tumour cells. As the lung is a major site for malignancy and infection and as there are large numbers of lymphoid cells including NK cells in the interstitial compartment of the lung, we evaluated the capacity of interleukin-2 (1L-2), a lymphokine capable of augmenting NK activity in vitro, to augment lung NK cell activity in vivo, usingdifferent routes ofIL-2 administration. We compared both systemic (i.v. and i.p.) and local (intrapleural and inhalation) routes of IL-2 administration (50000 UJdaily for 5 days) using CBA mice, assessing NK and LAK cell activity in the spleen (systemic) and in the lung. The target cells used for these studies were the YAC-1 (NK-sensitive) and P815, NO36 and HA56 (NK-resistant, LAK-sensitive) cell lines. Splenic NK activity was increased by 1.4-1 1.9-fold for i.v./i.p., respectively, compared with controls with both systemic routes of administration, and lung NK activity was increased 3-2-fold and 3-8-fold (i.v./i.p, respectively, P < 0.05), to levels which were comparable to systemic (splenic) NK activity following the same therapy. Intrapleural IL-2 administration similarly enhanced lung NK activity (3-3-fold) and splenic NK activity (1.3-fold; P<0.05 versus controls for both). Surprisingly, inhaled IL-2 suppressed both splenic and lung NK cell activity (84.8% and 78 ± 10% suppression, respectively, P<0.05). LAK cell activity was also enhanced in the lung by 1.8-8-fold in response to i.v., i.p. and intrapleural IL-2, whereas inhaled IL-2 was ineffective in generating LAK cell activity. These results suggest that the systemic and intrapleural administration of IL-2 effectively boost pulmonary NK and LAK activity whereas inhalation of IL-2 does not. Thus, in clinical situations where boosting of local lung NK or LAK cell activity is desired, these routes of IL-2 administration may be effective.
A novel strategy for targeting CD4+ PPD-reactive T cells against tumour cells using PPD monoclonal antibody heteroconjugates
Authors: MONTGOMERY A M P , WING M G , LACHMANN P J
Published: July 2010

We have constructed PPD monoclonal antibody heteroconjugates specific for a tumour-associated antigen of C57BL/6 melanomas or for human complement component C3d fixed de novo to murine fibrosarcoma cells (MC6A). The ability of our heteroconjugates to target CD4+ PPD-reactive T cells against the appropriate tumour targets was then determined in vitro. Heteroconjugate-treated B16-F10 and MC6A tumour targets were both able to present PPD to the specific T cells, resulting in activation and concomitant tymphokine secretion. Secreted lymphokines were then demonstrated to cause significant tumour cytolysis and cytostasis in vitro. Preliminary experiments in vivo suggest that this targeting system may provide the basis for a future immunotherapeutic strategy.
Titration of antibodies against neutrophil cytoplasmic antigens is useful in monitoring disease activity in systemic vasculitides
Authors: EGNER W , CHAPEL H M
Published: July 2010

Titration of antibodies against neutrophil cytoplasmic antigens (cANCA), as detected by indirect immunofluorescence. is shown to be clinically useful for monitoring disease activity in Wegener's granulomatosis and microscopic polyarteritis. Ten patients were followed (eight from presentation) prospectively for up to 2 years; during this lime there were six episodes of vasculitic relapse in four patients and five infective episodes and one pulmonary embolus in four patients. Titres of cANCA were markedly raised, both at presentation (1 /32–1/2048) and at vasculitic relapse (1/125–1/1048) but not in infection or embolism (negative. 1/16). Thus the titre of these antibodies can distinguish non-vasculitic illness from vasculitic relapse, in contrast to C-reactive protein levels which were raised in both. Titres of cANCA fell gradually after vasculitic relapse, in keeping with the half-life of IgG (3 weeks). C-reactive protein is a better measure of recovery.
Purification and characterization of IgG immunoconglutinins from patien with systemic lupus erythematosus: implications for a regulatory functiot
Authors: NILSSON B , EKDAHL K NILSSON , SVARVARE M , BJELLE A , NILSSON U R
Published: July 2010

The levels of IgG immunoconglutinins in plasma from patients with rheumatoid arthritis, systemic lupus erythematosus and primary biliary cirrhosis were monitored by ELISA. High levels of IgG immunoconglutinins were found mainly in plasma from patients with systemic lupus erythematosus. These immunoconglutinins bound to microtitre plate-fixed C3, C3b and C3c but poorly to C3d. This binding was inhibited by particle-bound C3b and iC3b but not by the corresponding soluble fragments. Furthermore, Western blot analysis revealed no immunoconglutinin-binding to reduced C3 peptides and no binding was shown to soluble C3 α and β chain by ELISA. IgG immunoconglutinins were purified from three plasma specimens by affinity chromatography on activated thiol sepharose ATS/C3 fragments. Two immunoconglutinin preparations that preferentially recognize ATS-C3b, inhibited C5-converlase function by 50–100% while one immunoconglutinin that recognized ATS-C3d,g had no effect. The two former immunoconglutinins also inhibited all three factor I cleavages in C3α chain but the latter inhibited only the third cleavage. None of the immunoconglutinins affected the binding of complement-coated anti-BSA/BSA complexes to CRI (CD 35) on human erythrocytes, but the two immunoconglutinins that inhibited all factor I cleavages also inhibited the factor I-induced release of anti-BSA/BSA complexes from CRI. The results show that immunoconglutinins recognize specific epitopes on bound C3 fragments and that they are able to modulate C3-mcdiated functions.
Anti-heparan sulphate reactivity in sera from patients with systemic lupus erythematosus with renal or non-renal manifestations
Authors: TERMAAT R M , BRINKMAN K , NOSSENT J C , SWAAK A J G , SMEENK R J T , BERDEN J H M
Published: July 2010

Previously, we have shown that anti-DNA can bind to heparan sulphate (HS). a constituent of the glomerular basement membrane (GBM). We hypothesized that binding of anti-DNA to HS in the GBM plays a role in the onset of systemic lupus erythematosus (SLE) nephritis. To test this hypothesis we measured the anti-HS reactivity in cross-sectional and longitudinal studies of SLE patients with or without nephritis. In the transverse serum study single serum samples from 26 SLE patients were studied. We found no correlation between anti-HS reactivity and previous development of nephritis (anti-HS positive: seven out of 16 with history of nephritis, two out of 10 without nephritis). However, six of the seven anti-HS positive sera in the nephritis group were obtained within 1 month of the onset of nephritis, suggesting a temporal relationship between anti-HS reactivity and onset of nephritis. In the longitudinal serum study between six and 16 serum samples were studied from each of 10 SLE-patients. In five out of five episodes of nephritis we found anti-HS reactivity before the onset or exacerbation of the nephritis. In four non-renal manifestations anti-HS reactivity was found in only one episode; in none of the three patients who remained clinically stable did scrum samples show anti-HS reactivity. Anti-HS reactivity was only found in sera positive for anti-DNA by Farr assay but the anti-HS titre was not a mere reflection of the reactivity measured in the Farr assay. This indicates that only a subpopulation of anti-DNA can bind to HS. We found a high correlation (r=0.99) between anti-HS reactivities in plasma and serum and we conclude that anti-HS reactivity in scrum samples from SLE patients is not due to in vitro complex formation during clotting. Although further prospective analysis is necessary, our data suggest that measurement of anti-HS reactivity in SLE patients might identify patients at risk for the development of nephritis.
Patients with myasthenia gravis and thymoma have in their sera IgG autoantibodies against titin
Authors: AARLI J A , STEFANSSON K , MARTON L S G , WOLLMANN R L
Published: July 2010

Patients with myasthenia gravis (MG) and thymoma have in their sera antibodies which react with non-receptor antigens from striated muscle. The purpose of this investigation was to characterize the antigen(s). Polypeptides in homogenates from rat skeletal muscle were separated by SDS-PAGE and trans-blotted to nitrocellulose. Sera from six patients with MG and thymoma stained a large (molecular weight >500 kD) polypeptide. while no staining was observed with sera from 20 non-thymoma MG patients. Titin is one of the large (> 500 kD) polypeptides of striated muscle and the antibody containing MG sera have antibodies that bind to titin in a preparation of myofibrillary proteins from rabbit skeletal muscle. The staining pattern is identical to that obtained with antiserum to titin, showing that the antigen has the same electrophoretic mobility as titin. Antibodies from the sera of the patients with MG and thymoma, affinity-purified on the large polypeptide, reacted with skeletal muscle sections in a cross-striational pattern, near the A/I band junction but within the I band, corresponding to the localization of one of the epitopes of titin. Our findings therefore indicate that the muscle antibodies found in the sera from some MG patients with thymoma are directed against titin.
Anti-mitochondrial antibodies (anti-M7) in heart diseases recognize epitopes on bacterial and mammalian sarcosine dehydrogenase
Authors: KLEIN R , BERG P A
Published: July 2010

The anti-mitochondrial antibody (AMA) anti-M7 has been shown to occur exclusively in sera from patients with acute and chronic myocarditis. Applying different enzymes of the inner mitochondrial membrane to ELISA, anti-M7-positive sera reacted only with sarcosine dehydrogenase (SD) from Pseudomonas aeruginosa. Testing these sera in the Western blot against a commercially available SD as well as against SD prepared from rat liver mitochondria, a determinant at 42 kD and 90 kD, respectively, was visualized. Using submitochondrial particles (SMP) from bovine heart and rat liver another major determinant at 64 kD could be observed with both antigen fractions. Liver SMP also expressed the SD-related, 90-kD epitope. Sera from patients with other AMA-positive and AMA-negative autoimmune diseases were negative with these different determinants. The identity of the 64-kD epitope on heart and liver SMP as well as the 42-kD polypeptide of bacterial SD and the 90-kD epitope on mammalian SD was proven by absorption studies and by elution of antibodies from the antigen bound to the immobilon sheets after immunoblotting. The SD enzyme activity was not affected by anti-64-kD and anti-42-kD antibodies in vitro. It is concluded that anti-M7 antibodies may be stimulated by an antigen expressed on cardiocytes during an infection which shares epitopes with SD, an evolutionary highly conserved protein. SD-sensitized B cell clones could therefore be triggered by the M7-antigen which shows homology to SD.
The expression of γΔ T cell receptor and the prevalence of primed, activated and IgA-bound T cells in Behçet's syndrome
Authors: FORTUNE F , WALKER J , LEHNER T
Published: July 2010

Mucosal ulceration of the oral, and to a lesser extent genital tissues is an essential feature of Behçet's syndrome and is associated with changes in the IgA class of immune responses. Indeed, a significant increase in the proportion of cytophilic IgAl was found in circulating CD8 and CD4 cells (P < 0.01), with a corresponding decrease in IgA-Fc receptors on these T cells. Furthermore, 30–40% of the cytophilic IgA1 on T cells may have been of the polymeric secretory type and the rest of the monomeric variety. IgA isotype of B cells was also significantly increased (P < 0.001), without an overall change in circulating B cells. However, a surprising finding was the significant up-regulation of γŞ T cell receptor in the CD8 (P < 0.01) in the absence of a change in the proportion of αβ T cell receptor. The results suggest that some common microbial antigen might initiate at the mucosal surface an immune defence reaction characterized by T cells with γΔ receptors and IgA-specific B cells. However, IgAl bound to circulating T cells may down-regulate the central T cell function.
Terminally differentiated human intestinal B cells. J chain expression of IgA and IgG subclass-producing immunocytes in the distal ileum compared with mesenteric and peripheral lymph nodes
Authors: BJERKE K , BRANDTZAEG P
Published: July 2010

Two-colour immunofluorescencc staining for intracellular J chain and IgA (or J chain and IgG) was performed on tissue sections of normal human ileal mucosa (eight adult kidney donors), mesenteric lymph nodes (MLN), peripheral lymph nodes, and palatine tonsils. The most prominent J chain positivily was seen for IgA (97.3%) and IgG (81.7%) immunocytes in the ileal lamina propria (LP). Moreover, the proportion of J chain-expressing exlrafollicular immunocytes was significantly higher (P < 0.05) in MLN than in peripheral lymph nodes for the IgA class (58.5%versus 25.6%); the same proportion for the IgG class was 45.9%versus 30.4%. In clinically normal palatine tonsils of adults, extrafollicular J chain expression was much lower than in peripheral lymph nodes; 14.2% for IgA cells and 5.5% for IgG cells. When related to subclass production, J chain expression was found to be higher for IgA2 than for IgA 1 cells in all tissues examined (palatine tonsils excluded because of a small number of lgA2 cells), the difference being significant in M LN and ileal LP (P < 005). The J chain positivity tended to be higher for all IgG subclasses in MLN than in peripheral lymph nodes; this difference was significant (P <005) for lgG2-producing immunocytes. Taking J chain expression as a marker of clonal immaturity, our results may reflect to some extent distribution of newly generated memory B cell clones from gut-associated lymphoid tissue to MLN. peripheral lymph nodes, and palatine tonsils in a strikingly decreasing order.
FK506, a novel immunosuppressive agent, induces antigen-specific immunotolerance in active Heymann's nephritis and in the autologous phase of Masugi nephritis
Authors: OKUBO Y , TSUKADA Y , MAEZAWA A , ONO K , YANO S , NARUSE T
Published: July 2010

FK506 is a new drug which has potent immunosuppressive activity. We studied its immunosuppressive effects on active Heymann's nephritis and the aulologous phase of Masugi nephritis. The induction of active Heymann's nephritis was completely suppressed by FK506 injected simultaneously with the antigen (day 1) and then daily for 14 days at a dose of 0.64 mg/kg per day or more. With a lower dosage of this agent, antibody production and immune deposits in the glomerular basement membrane occurred despite the suppression of proteinuria. Similar results were obtained in rats on other treatment schedules (1–7 days or day 8–14 days duration). Rats that were prevented from developing Heymann's nephritis or the autologous phase of nephrotoxic antiserum nephritis by FK506 treatment exhibited a suppressed immune response to a second immunization of the same antigen even 4 weeks after cessation of drug administration; however, they developed antibodies when inoculated with other antigens. Rat peripheral leucocyte counts and scrum creatinin were not remarkably influenced by the administration of FK506. These results indicate that FK506 has potent immunosuppressive activity, and it is suggested that it is able to induce an antigen-specific immunotolerance.
Anti-DNA idiotype- and anti-idiotype-specific T cell responses in patients with systemic lupus erythematosus and their first-degree relatives
Authors: MENDLOVIC S , SEGAL R , SHOENFELD Y , MOZES E
Published: July 2010

We examined the proliferative responses of T cells of patients with systemic lupus erythematosus (SLE). their first-degree relatives, and healthy donors, to a human monoclonal antibody that bears a common anti-DNA idiotype, 16/6 Id, and to a nurine, 16/6 Id-specific, monoclonal antibody. Both 16/6 Id+ and 16/6 Id-specific antibodies were previously shown to be involved in the induction of experimental SLE in mice. Here we show that T cells of fewer SLE patients, as compared with healthy donors, could proliferate to both antibodies. The difference between T cell responses of patients and controls to the 16/6 was found to be significant. The proliferative responses of T cells of first degree relatives of SLE patients to the anti-16/6 Id were found to be significantly lower compared with the responses detected in healthy donors and in SLE patients. The responses of T cells of SLE relatives to the 16/6 Id were also found to be lower than those of healthy donors, but this difference was not significant. The present study suggests a possible involvement of T cells, and specifically of idiotype and anti-idiotype specific T cells, in SLE.
Persistence of allospecific helper T cells is required for maintaining autoantibody formation in lupus-like graft-versus-host disease
Authors: ROZENDAAL L , PALS S T , GLEICHMANN E , MELIEF C J M
Published: July 2010

Induction of a graft-versus-host (GVH) reaction (GVHR) in non-irradiated (C57BL/10ScSn × DBA/2)F1 mice (BDF1) with DBA/2 lymphoid cells leads to chronic GVH disease (GVHD). One of the pathological alterations of this type of GVHD is hyperplasia of host B cells with production of lupus-like autoantibodies. This hyperstimulation of host Bcells has previously been demonstrated to be induced by alloreactive donor T helper cells that were also proposed to maintain it. We provide three pieces of experimental evidence in support of this concept. First, treatment of mice with chronic GVHD by injection of monoclonal anti-Thy-1.2 antibodies, performed at week 6 after the injection of C57BL/6 lymphoid cells into (C57BL/6 × C57BL.bml2)F1 mice led to a significant decrease in the litre of anti-nuclear antibodies. Second, CD4+ donor T cells persisted in BDF1 mice with GVHD (GVHF1) for at least 10 weeks after the induction of GVHR; these T cells showed alloreactive helper activity against H-2b MHC determinants of the opposite parent in vitro. Third, T cells of GVH F1 mice, obtained 2 months after the induction of GVHR and transferred into normal secondary recipients, induced signs of chronic GVHD in DBF1 but not in DBA/2 mice. The combined results show that persisting donor T helper cells in GVHF1 mice retain their alloreactivity towards H-2 class II antigens for a long time after the induction of GVHR and they strongly suggest that these T cells are also the driving force behind the production of lupus-like autoantibodies at the late stage of chronic GVHD.
A T cell receptor beta chain polymorphism is associated with patients developing insulin-dependent diabetes after the age of 20 years
Authors: McMILLAN S A , GRAHAM C A , HART P J , HADDEN D R , McNEILL T A
Published: July 2010

We have studied the BglII polymorphism near the T cell receptor beta chain constant region (TcR-Cβ) gene, HLA-DR genotypes and certain autoimmune features in 102 patients with type I (insulin-dependent) diabetes. There was a significant decrease in the frequency of the 1:1 genotype (P= 0.008) and an increase in the 1:2 genotype (P= 0.03) of the BglII TcR polymorphism in the group of patients who developed type-I diabetes after the age of 20 years. This group of patients also showed an increased incidence of autoantibodies (especially islet cell antibody), a family history of diabetes and the presence of other autoimmune diseases. The frequency of this polymorphism in patients who developed type I diabetes before the age of 20 years was similar to a non-diabetic group. These results suggest that there are two genetically distinct groups of patients with type I diabetes. HLA-DR3 and HLA-DR4 genotypes were also increased in the diabetic patients but no significant difference was observed between HLA-DR genotypes, the TcR-Cβ genotypes, the age of diagnosis or with other autoimmune features. Patients developing type I (insulin-dependent) diabetes after the age of 20 years have an additional genetic susceptibility for diabetes associated with the TcR-Cβ gene.
A delayed-type hypersensitivity skin-test system using the insulinoma cell line RINm5F to monitor beta cell-specific cellular autoimmune reactivity in the spontaneously diabetic BB/O rat
Authors: VOORBY H A M , JEUCKEN P H M , DREXHAGE H A
Published: July 2010

Islet-specific autoimmune reactivity (humoral and cell-mediated) is the basis for the insulitis process of type I diabetes mellitus. In this report a delayed-type hypersensitivity (DTH) skin test was used to monitor the presence of an islet-specific cell-mediated autoimmune component in BB/O rats. The BB/O rat is a strain characterized by the spontaneous development of type I diabetes. Intact RINm5F cells as well as a RINm5F cell membrane preparation were used as DTH skin test antigens. Rats of different ages and disease stages were tested in the ear with the insulinoma cell line and its cell membrane preparation. As control antigens, the fibroblast cell line 3Y1 and a cell membrane preparation made thereof were used. The DTH reaction system showed a positive cell-mediated reactivity in BB/O rats for membrane-bound RINm5F cell antigens, and not for the control fibroblast 3Y1 cell membrane determinants. The true DTH character of the skin test was established by the time-course of the reaction (maximum at 24 h), the histopathology (infiltration by dendritic cells, lymphocytes and macrophages), and the possibility to transfer the reaction with spleen cells and lymph node cells. The DTH test towards RINm5F cells showed the highest prevalence of positivity (l00%) in BB/O rats around the onset of diabetes (3 weeks before to 3 weeks after the onset of glucosuria). The prevalence of DTH positivity was 56% in the period of more than 3 weeks before the onset of glucosuria. In BB/O rats with a duration of glucosuria of more than 3 weeks, the prevalence of positivity was around 60–70%).
Immune responses to fractionated cytomegalovirus (CMV) antigens after HIV infection. Loss of cellular and humoral reactivity to antigens recognized by HIV, CMV+ individuals
Authors: CONVERSE P J , FEHNIGER T E , EHRNST A , STRANNEGÅRD Ö , BRITTON S
Published: July 2010

In order to delineate the molecular pathogenesis of the increased susceptibility to CMV disease in HIV infection, the patterns of antigen responsiveness in HIV-infected and non-infected individuals were investigated. CMV was fractionated by SDS-PAGE and electroblotted onto nitrocellulose. Lymphoproliferative responses of healthy HIV–, CMV+ individuals and HIV+, CMV+ asymptomatic patients to a whole CMV antigen preparation and to 20 fractions of nitrocellulose-bound CMV were then compared. Three fractions of approximate molecular weight of 130–165, 65–75, and 55–65 kD appeared to contain the major T cell stimulating antigens for HIV, CMV– individuals. A statistically significant depression of responses to fractions containing antigens in the ranges of 130–165 kD and 55–65 kD but not to whole CMV was seen in HIV+ individuals compared with controls. In healthy controls, the sum of the proliferative responses as measured by 3H-thymidine uptake to these three major fractions was approximately equal to the response to a whole CMV antigen preparation, whereas it was less than half of this response in five out of six HIV+ subjects. When antibody activities to CMV antigens were analysed by immunoblotting of sera from the two subject groups and also sera of ARC and AIDS patients, a selective loss of reactivity was revealed in 10 out of 19 HIV+ subjects to a band of 26–28 kD whereas all 15HIV–, CMV+ controls recognized this band. Serum IgG and IgM values were both significantly higher in HIV+ individuals than in controls. These findings suggest that specific lesions in the repertoire of immune responsive CMV antigens occur in HiV+ individuals.
Changes in the serum concentration and the glycosylation of human αl-acid glycoprotein and αl-protease inhibitor in severely burned persons: relation to interleukin-6 levels
Authors: POS O , VAN DER STELT M E , WOLBINK G-J , NUSTEN M W N , VAN DER TEMPEL G L , VAN DIJK W
Published: July 2010

The relation between interleukin-6 (TL-6) levels and changes in serum concentrations and glycosylation (concanavalin A affinity) of two human acute-phase glycoproteins, αl-acid glycoprotein (AGP) and α1-protease inhibitor (PI), was studied in sequential serum samples of burn patients. The level of IL-6 was already increased at the first day following injury, and after a dip at day 2 or 3 rapidly reached a second maximal value at day 4 or 5. The serum concentrations of AGP and PI reached their maximal values after day 5 and remained at a high level throughout the total period studied (7 weeks). The concanavalin A reactivities of both acute-phase glycoproteins were found to be elevated only during the first 2–25 weeks. Maximal values were observed on day 2 and from day 7 to 16, following closely the rise and fall of the lL-6 serum level. After day 16, the concanavalin A affinity rapidly declined long before a decrease was observed in the serum concentrations of AGP and PI. Our previous in vitro studies have indicated an involvement of lL-6 in the induction of both secretion and increased concanavalin A affinity. This study indicates that IL-6 could play a causal role in the induction of both phenomena in vivo.
Suppression of lymphokine-activated killer (LAK) cell induction mediated by interleukin-4 and transforming growth factor-β1: effect of addition of exogenous tumour necrosis factor-alpha and interferon-gamma, and measurement of their endogenous production
Authors: BROOKS B , CHAPMAN K , LAWRY J , MEAGER A , REES R C
Published: July 2010

Recombinant human interleukin-4 (rhlL-4) and transforming growth factor-βl (TGF-β1) suppressed the induction of lymphokine-activated killer (LAK) activity induced by recombinant human interleukin-2 (rhIL-2) in peripheral blood lymphocytes. DNA synthesis and the expression of the p55 alpha chain of the IL-2 receptor (Tac antigen) were also inhibited. The inhibitory effect was greatest when these factors were added during the first 48 h of a 4-day culture, with reduced cytolytic activity against both natural killer (NK) resistant and NK-sensitive tumour cell line targets. The suppressive action of both cytokines was accompanied by a reduction in tumour necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) levels in lymphocyte culture supernatants. Recombinant human IFN-γ (rhIFN-γ). but not recombinant human TNF-α (rhTNF-α) was able to overcome the inhibitory effect of recombinant human interleukin-4(rhIL-4) on LAK induction and DNA synthesis but not Tac antigen expression. However, cytotoxicity induced by rhI FN-γ alone was also suppressed by rhIL-4 and TGF-β1, inferring that rhIFN-γ-mediated abrogation of rhlL-4 suppression was not simply a direct IL-2-indcpendent effect on cytotoxicity. In addition, rhIL-4 did not increase TGF-β production from rhIL-2-activated peripheral blood mononuclear cells, suggesting that rhIL-4 did not mediate reduction of rhIL-2 responses through the induction of TGF-β release.
Preparation of anti-T idiotype monoclonal antibody reacting with human T leukaemic cell lines and with a small percentage of peripheral T lymphocytes
Authors: TAKAHASHI T , IMAI K , SUGIYAMA T , SASAKI T , YACHI A
Published: July 2010

Monoclonal antibody (MoAb) KT38 raised against a human T leukaemic cell line TALL-1, reacted with another T leukaemic cell line Jurkat, but not with any other cell lines tested. The co-modulation of CD3 and KT38 antigen was observed with stimulations or either MoAb T3 or KT38 on both TALL-1 and Jurkat. Upon radioimmunoprecipitation and SDS-PAGE analysis, MoAb KT38 precipitated the heterodimer of 40–60 kD from Jurkat and TALL-1 under reducing conditions. Thus, MoAb KT38 is considered to be an anti-T idiotype (Ti) antibody to TALL-1 and Jurkat cells. MoAb KT38 was also shown to react with a minor population of peripheral blood lymphocytes (PBL) and with very few cells (0.5–2.0%) in the paracortical area of the lymph node. When PBL were stimulated in a KT38-coated culture flask for 5 days, the percentage of KT38-positive PBL was markedly increased. The CD3 antigen on these cultured PBL in the flask was modulated by the stimulation with MoAb KT38. Thus, it is suggested that a common idiotype exists on the T cell receptor of Jurkat, TALL-1 and a small percentage (1.9–6.1%) of PBL.