bluuebunny/crossref_metadata_2025 huggingface dataset:
The Development of IgA‐Specific Antibodies to Escherichia coli O Antigen in Children
Authors: GLEESON M. , CRIPPS A. W. , CLANCY R. L. , WLODARCZYK J. H. , DOBSON A. J. , HENSLEY M. J.
Published: June 2006

One hundred and sixty‐five infants were studied longitudinally from birth to 5 years of age. One hundred and twenty‐three school‐age children and 27 adults were examined cross‐sectionally. Total salivary IgA levels and IgA antibodies against Escherichia coli O antigens were measured. Total IgA levels were low (<20 mg/l) from birth to 4 years of age. At 5 years of age there was a dramatic increase in the total IgA level (geometric mean = 100.7 mg/l), after which the levels fell to values similar to those observed in adults (adult geometric mean = 53.2 mg/l). Low levels of IgA‐specific E. coli antibodies were observed for the first 4 years of life (<1.0 ELISA units). There was a gradual increase in specific antibodies between 5 and 9 years of age (geometric men at 9 years=4.66 ELISA units) to levels similar to those observed in adults (adult geometric mean=8.20 ELISA units). It is suggested that the patterns of development for these variables reflect a balance between antigenic exposure and immune control mechanisms.
Mitogen Stimulation Promotes Human T Lymphocyte Adhesion to Fibronectin
Authors: KURKI P. , VARTIO T. , VIRTANEN I.
Published: June 2006

Purified human peripheral blood T lymphocytes were shown to adhere to growth substrata coated with purified human plasma fibronectin (pFn) and its M1 120,000‐140,000 proteolytic fragments containing the cell‐binding site. In contrast, significant binding to laminin‐ or type I collagen‐coated surfaces could not be demonstrated. Binding of T cells to pFn could be inhibited by the synihetic peptidc Arg‐Gly‐Asp‐Ser. Activation of T lymphocytes with concanavalin A (Con A) and a phorbol ester. I2‐O‐tetradecanoyl‐phorbol‐13‐acetate (TPA), led to a higher adherence to pFn than in unstinuilaled, resting T cells. Activation with only Con A in the presence of accessory cells also promoted binding. Increased adherence of T cells to pFn could be demonstrated as early as 2 h after the onset ol stimulation and reached its maximum in 2‐3 days. Furthermore, activated but not resting T cells actively spread on pFn‐coated surfaces and displayed an altered F‐actin organization. In an overlay assay of electrophoretically separated polypeptides of activated T lymphocytes, pFn bound to a high molecular weight polypetide of M, 190,000. suggesting that the cells bind lo pFn via a receptor‐like molecule. Thus, adhesion of pFn may be a two‐stage process. At the first stage cells bind to Fn. Activated T cells proceed to the second stage, where cells begin to spread on pFn. This may be due lo an altered relationship between Fn receptors and microfilaments.
Analysis of the Antigenic Profile of Mycobacterium leprae: Cross‐Reactive and Unique Speeificities of Human and Rabbit Antibodies
Authors: EHRENBERG J. P. , GEBRE N.
Published: June 2006

Thirty‐two mycobacterial components were detected by antibodies contained in leprosy patients' sera across the clinical spectrum and rabbit anti‐M. leprae hyperimmune sera by western blot analysis of armadillo‐derived M. leprae antigen preparations. Sera of borderline tuberculoid patients were found to contain antibodies recognizing 18 M. leprae components. While the reactivity of the sera on the lepromatous pole seemed to be distributed oven the entire molecular weight range, most of the reactivity in the borderline tuberculoid patients was directed at higher molecular weight components (>70,000). Identification of a series of previously unrecognized M. leprae components offers new possibilities in regard to the potential use of these antigens as targets for immunodiagnosis. Antibodies contained in The rabbit anti‐M. leprae sera reacted with 19 M. leprae components. Antigens migrating at 64,000, 38,000, and 22,000 were detected by the rabbit seraonly. Evidence of extensive cross‐reactivity between M. leprae and BCG organisms emphasizes the need to use well‐characterized antibody probes to determine the specificity of select mycobacterial antigens. The potential usefulness of rabbit monospecific hyperimmune sera to select M. leprae fractions in immunodiagnosis, in immune regulation studies, or as a tool to screen for mycobacterial products in lambda gt11 phage lysates of E. coli is discussed. Select M. leprae components were partially purified and their recovery assessed through SDS‐PAGE analysis of Coomassie blue‐stained gels.
Diminished Natural Killer Activity in Pregnancy: Modulation by Interleukin 2 and Interferon γ
Authors: VAQUHR S. , DE LA HERA A. , JORDÁ J. , MARTÍNEZ‐A. C. , ESCUDERO M. , ALVAREZ‐MON M.
Published: June 2006

We investigated the alterations of natural killer (NK) cells in pregnancy, which led to decreased killing from the first trimester to the puerperium. We show that this phenomenon cannot be ascribed to a defective numher of NK cells since the amounts of HNK‐1+, CD16+ (Leu 11), and CD11b+ (OKMl) cells were within normal ranges. Recombinant interleukin 2 (rIL‐2) corrects the functional defect in a dose and time‐dependent manner, without modification in the surface phenotype of the population. Analysis of the pattern of target cell susceptibility to lysis, together with ihe similar ability of recombinant interferon γ (rlFN‐γ) to correct the deficiency, and CD16+, CD3− phenotype of the precursor and effector lymphocytes, demonstrated that the induced cytotoxicity was mediated by NK cells. Inhibitors in pregnancy sera block IL‐2 production by specific T cells, hut we show that they do not influence either NK activity or its reconstitution hy rlL‐2. These findings place the deficiency at the level of NK cell maturation into cytotoxic effector lymphocytes. Thus, homeostasis of the NK activity of pregnant woman may provide a biological model for clarifying the internal mechanisms regulating NK‐cell activation in vivo. Present studies in vitro suggest that modulation of lymphokine production may play a role in the adaptive response of NK cells in pregnancy.
Production of Prostaglandin E2 and Interleukin 1 by Mouse Peritoneal Macrophages Stimulated with β‐l, 3‐D‐Glucan Derivatized Plastic Beads
Authors: RASMUSSEN L.‐T. , UPSKY P. E. , SELJELID R.
Published: June 2006

Fluorescein‐labelled plastic microbeads, with or without covalently attached β‐l, 3‐D‐glucan, were injected into the peritoneal cavity of mice. Peritoneal cells were subsequently analysed by flow cytometry according to fluorescence and light scatter and separated into fluorescence positive and negative cells. We report that cells from animals treated with glucan‐plastic beads produced large amounts of prostaglandin E2 (PGE2) whether the cells actually contained heads or not. On the other hand, cells from animals treated with glucan‐plastic beads produced less thymocyte‐stimulatory activity—presumably corresponding to interleukin 1 (IL‐l)—than cells from control animals treated with commercial latex beads. However, when indomethacin was added, either in vivo or in vitro, cells from animals treated with glucan‐plastic beads produced more thymocyte‐stimulalory activity than controls. We interpret this to mean that glucan‐plastic beads stimulate both IL‐l and PGE2, but that under circumstances where the cellular cyclo‐oxygenase is not inhibited, the PGE2 will block IL‐1 production.
Remarkable Depression of CD4+2H4+ T Cells in Severe Chronic Active Epstein‐Barr Virus Infection
Authors: FRANCO E. , KAWA‐HA K. , DOl S. , YUMURA K. , MURATA M. , ISHIHARA S. , TAWA A. , YABUUCHI H.
Published: June 2006

In order to better understand the features of chronic active Epstein‐Barr (EB) vims infection, we employed two‐colour immunofluorescence staining with monoclonal antibodies and flow cytometry analysis to study the lymphocyte phenotypes of two patients with severe symptoms of this disorder as well as four patients with mild symptoms. We found an increased number of activated T cells, as characterized by CD4+ la+, CD8+Ia+, or CD4+Tac+ phenotypes, and a markedly decreased CD4+2H4+ T cell subpopulation, previously characterized as a suppressor‐inducer subset, in the patients with severe symptoms. In contrast, the four patients with mild symptoms showed only a slightly elevated number of activated T cells and a normal CD4+2H4+/CD4+ ratio. These phenotypic differences may suggesst heterogeneity in this disorder. Also, a failure in ihe suppressor‐inducer population could contribute to changes in the host‐virus relationship and the degree of the decrease in this population may correlate directly with ihe severity of the disease.
Accessory Cell‐Dependent T‐Cell Activation via Ti‐CD3
Authors: HALVORSEN R. , LEIVESTAD T. , GAUDHRNACK G. , THORSBY E.
Published: June 2006

The activation of resting T cells to interleukin 2 (IL‐21) production and DNA synthesis via T1‐CD3 is dependent on accessory cells (AC) using positively selected, resting I cells activated with particle‐bound anti‐CD3, we investigated the ability of various cell lines to function as AC. We found that cell lines able to act as AC all expressed LFA‐3, while cell lines not expressing LFA‐3 were unable to provide AC signals. This applied to CD3+. CD4+, and CD8+ T cells. Sheep red blood cells (SRBC), which express LFA‐3‐like molecules, also had a weak, hut significant AC function in this test system. Both CD4+ and CDS+ T cells activated with particle‐bound anti‐CD3 could be induced to enter DNA synthesis in the absence of AC when monoclonal antibodies reacting with CD2 were present instead of AC IL‐2 production could be detected in the latter cultures but not when positively selected CD3+ or CD2+ T cells were cultured alone. Our data suggest that activation of resting T cells via CD3 will lead to IL‐2 receptor expression, while the interactions between LFA‐3 and its ligand CD2 provide the necessary secondary signals for IL‐2 production and induction of DNA synthesis
Thymic Accessory Cells Required for the Increased Responsiveness of Thymocyte Subpopulations to Interleukin 1‐Like Monokine in Guinea Pigs
Authors: AIDA Y. , KAKIMOTO K. , KAWAKAMI K. , CHIBA J. , AONO M. , ONOUE K.
Published: June 2006

The proliferative response of guinea‐pig thymocytes to co‐mitogenic stimulation with phytohaemuglutinin and the guinea‐pig interleukin 1 (IL‐1)‐like lymphocyte‐activating monokine was lost by removing the cells that adhere to a Sephadex G‐10 (G‐10) column or the cells of low density in a Ficoll‐Conray gradient. The diminished response in the G‐10 non‐adherent thymocyte or high‐density thymocyte fraction was restored by the addition of a macrophage‐depleted 6–10 adherent thymocyte fraction or a low‐density. Ia‐positive thymocyte fraction but not by the addition of peritoneal macrophages. These results suggest that the accessory cells which mediate the increased responsiveness of thymocytes to the IL‐1‐like monokine existed in G‐10 adherent cell fractions and the cells with this accessory function were not macrophages The accessory cells were shown to DC of low density, glass‐non‐adherent, G‐10‐adherent, Fc receptor‐negative, and Ia‐positive. These results also suggest that the G‐10‐non‐tidherent and high‐density thymocyte subpopulation, which is unresponsive or responds very little to the IL‐l‐like monokine by itself, acquires responsiveness to the monokine and proliferates by Stimulation with the 1L‐1‐Iike monokine and lectin in the presence of the accessory cells.
Induction of Basophilic Differentiation in the Human Basophilic Cell Line KU812
Authors: ALMLÖF I. , NILSSON K. , JOHANSSON V. , ÅKERBLOM E. , SLOTTE H. , AHLSTEDT S. , MATSSON P.
Published: June 2006

The potential for differentiation of the human basophilic leukaemia cell line KUS12 was examined by means of a panel of physiologic and non‐physiologic substances used as inducers. The phenotypic characteristics of non‐induced KU812 cells included an immature morphology with scanty cytoplasmic granulation, expression of a low amount of high affinity, but no low affinity receptors (CD 23) for IgE, and a capacity for low‐rate histamine synthesis. The differentiation process was characterized by a rapid (24 h) increase in histamine production, a slower morphological maturation with the development of Alcian blue stainable granula demonstrable after 72 h. Concomitant with the phenotypic alterations, cell growth was inhibited. Differentiation in KU812 cells was inducible by Ara‐C and to some extent by sodium butyrate, but not by dimethyl sulphoxide, retinoic acid, or gamma‐interferon. Conditioned medium (CM) from cultured peripheral blood cells from atopic individuals and 18 out of 22 analysed glioma cell lines induced differentiation of the KUS12 cells, whereas supernatant from only 1 out of 21 other cell lines, including carcinoma, melanoma, sarcoma, leukaemia, and normal fibroblasts had this activity. CM from the T‐leukaemic cell line. Mo, also induced KU812 differentiation. A primary fractionation of the active substance from this cell line by reversed phase chromatography eluted the active substance at a concentration of 42–44% acetonitrile. Our present study has shown that the KU812 may serve as an appropriate model to study differentiation of basophils. In addition, its fast and specific response to biological factors makes it suitable as a biological assay for determination of active factor produced by atopic individuals.
T‐ and B‐Cell Functions in IgA‐Deficient Patients
Authors: KLEMOLA T. K. , ESKOLA J. , SAVILAHTI E.
Published: June 2006

In vitro lymphocyte function of 60 IgA‐deficient patients(IgAdp) were studied. In the mitogen‐induced lymphocyte activation test, peripheral blood mononuclear cells (PBMC) of IgAdp showed a weaker response to pokeweed mitogen (PWM) than those ill controls (P<0.05), and the responses to phytohaemagglutinin (PHA) and concanavalin A (Con A) were normal. The amounts of IgA, IgG, and IgM secreted by PBMC from controls and IgAdp were measured in vitro by enzyme immunoassay. B cells were activated by PWM with or without hydrocortisone (HC) to inhibit HC‐sensitive suppressor cells. Lymphocytes of IgA‐deficient patients synthesized only minute amounts of IgA in vitro and the amounts of IgA synthesized correlated well with the serum IgA levels. Both IgG and IgM secretion by PWM‐Stimulated PBMC of IgAdp were also subnormal. The lymphocyte subset analysis in the peripheral blood revealed normal numbers and ratios of T cells, but IgAdp had reduced percentages of surface IgA‐bearing cells. Co‐culture experiments with isolated B cells. CD4+ and CD8+ cells from IgAdp, and controls showed low B‐call capacity for IgA production as the most constant finding Defects in T‐cell functions or chances in the proportions of surface antigen‐carrying lymphocytes (other than IgA+) were infrequent and were not associated with any particular group of patients or type of IgAd (primary, acquired, or transient).
Direct Evidence for Circulating apoSAA as the Precursor of Tissue AA Amyloid Deposits
Authors: TAPE C. , TAN R. , NESHEJM M. , KISILEVSKY R.
Published: June 2006

A precursor product study was carried out using tritiated, undenatured murine high‐density lipoprotein (HDL)‐apoSAA, to assess whether circulating HDL‐apoSAA is the precursor of tissue AA amyloid deposits. The quantity of label accumulating in AA over a 24‐h period was determined per unit weight of spleen. In addition a mathematical assessment of the quantity of label that should have accumulated in splaenic AA within 2–1 h was nude, based on the half‐life of circulating apoSAA, the rate of change of the specific activity of circulating apoSAA, and rate of deposition of splenic AA. The observed result was approximately 47%, of the theoretically predicted value. The latter did not include extraction efficiencies of AA peptide from spleen nor losses which might nave occurred during column fractionation of splenic extracts The observed and predicted results are therefore in remarkably good agreement and indicate that circulating apoSAA is the major source of protein for splenic AA amyloid deposition.
Elimination of CD8+ T Cells in Vivo Does Not Break Induced Immunospecific Tolerance to Experimental Allergic Neuritis in Rats
Authors: STRIGÅRD K. , OLSSON T. , LARSSON P. , HOLMDAHL R. , HÖJEBERG B. , KLARESKOG L.
Published: June 2006

The role of CD8+ T ‘cytotoxic/suppressor’ T cells in induced immunospecific tolerance and during recovery after actively induced disease was examined by means of elimination of CD8+ cells from Lewis rats using in vivo treatment by Ox8 monoclonal antibodies, in experimental allergic neuritis (EAN). Animals depleted of CD8+ T cells alter recovery from EAN did not show any clinical signs of relapse. Other animals were pretreated with the peripheral nerve basic protein P2 and thereby rendered resistant to disease induction with a potentially neuritogenic emulsion. The elimination of CD8+ T cells did not result in EAN here either. Thus, the CD8+ T‐cell population does not seem to participate in the suppression of this autoimmune disease under these experimental conditions.
The Expression of CR1 and CR3 on Non‐Modulated and Modulated Granulocytes of Healthy Blood Donors as Measured by Flow Cytofluorometry
Authors: HED J. , BERG O. , FORSLID J. , HALLDEN G. , LÄRKA‐RAFNER G.
Published: June 2006

Complement receptors, CR1 and CR3, on neutrophils increase their cellular spontaneously at 37°Cor after mechanical stress during the cell preparation. We have established a cell preparation procedure and a cytofluorometric immunoassay method to evaluate the receptor expression in vivo in this study. The expression of CRI and CR3 was studied after haemolysis in NH4C1 at different temperatures and incubation intervals. It was shown that cell preparation and receptor analysis must be performed at 15°C or lower to avoid up‐regulation of the receptor structures. Two minutes' Incubation at 20°C was sufficient to modulate the cells in this regard. Granulocytes from healthy blood donors were analysed and the mean fluorescence intensity (MF1), which reflects the number of receptors on (he ceil surface, showed a normal distribution for the CRI (n=158) and CR3 (n=76) expression in the healthy population. The MFI of the two receptors showed a correlation (r=0.71). Granulocytes from all donors could be modulated at 37°C to a similar degree for both receptors (r=0.76), despite the fact that they are supposed to be mobilized from different intracellular pools. A group of patients with localized inflammatory processes, such as sinusitis, differed from the healthy controls. In this group, the CR1 expression was not normally distributed, but IS out of 26 patients (58%) had granulocytes with a CR1 expression exceeding the mean±SD of the normal population.
Oestrogen and Human T Lymphocytes: Presence of Specific Receptors in the T‐Suppressor/Cytotoxic Subset
Authors: STIMSON W. H.
Published: June 2006

Human spleen cells were investigated for specific oestrogen binding using oestradiol‐17β. High affinity, low capacity binding was observed (Kd=2.5±10−10m) and the characteristics of the oestrogen receptor identified were typical of this type of species. T lymphocyte subpopulations were prepared by negative selection by means of the monoclonal antibodies Leu 2a and 3a. Oestrogen receptors were only found in T cells of the suppressor/cytotoxic (CD8‐positive) subset while helper/inducer lymphocytes (CD4‐positive) showed no significant steroid binding by cytosol and nuclear assays. These results are supported by data from experiments into the effects of oestradiol‐17β on immunoglobulin secretion, where it was apparent that only the suppressor/cytotoxic T‐cell subpopulation was responsive to the presence of the steroid.
The Influence of T Cells on Homogeneous Immunoglobulins in Sera of Athymic Nude Mice during Aging
Authors: VAN DEN AKKER T. W. , TIO‐GILLEN A. P. , SOLLEVELD H. A. , BENNER R. , RADL J.
Published: June 2006

In this study, results are presented which are in agreement with predictions made on basis of the ‘three‐Stage hypothesis’ on the development of benign monoclonal gammapathy (BMG). In a T‐cell depletion model, C57BL/Ka nude mice were shown to develop single and multiple homogeneous immunoglobulins (H‐Ig) during aging in the highest frequencies known so far. Ninety percent of the C57BL/Ka nude mice displayed one or more H‐Ig at 12 months of age. In a T‐cell supplementation model, infusion of corticosteroid resistant T cells into 9‐month‐old BALB/c nude mice resulted in a decrease in the Frequency of H‐Ig from 43% at 9 months down to 20% at 15 months of age. In contrast, the frequency of H‐Ig in the control group increased from 40% at 9 months up in 68% at 12 months. The results show that normally functioning T cells are essential for the generation of a normal, heterogeneous Ig spectrum; they further support the validity of the three‐stage hypothesis with regard to the role of an impairment of the T immune system in the pathogenesis of BMG.
Anti‐Idiotypic Antibody Used for the Localization of Parenterally Administered Monoclonal Anti‐Progesterone Antibody in Mice
Authors: WANG M.‐W. , HEAP R. B. , KING I. , TAUSSIG M. J. , WHYTE A.
Published: June 2006

Affinity‐purified rabbit and sheep anti‐idiotypic antisera raised against mouse monoclonal anti‐progesterone IgG1 antibody (DB3) or mouse myeloma IgG1 protein P3 (MOPC 21) showed high binding specificities to the respective idiotypes used for immunization as determined by RIA or ELISA. They have been used in an indirect immunofluorescent method to demonstrate the localization of parenterally administered idiotypes in pregnant or pseudopregnant BALB/c mouse frozen tissue preparations, at known stages post coitum after a single intraperitoneal or intravenous injection of DB3 or P3, DB3 was visualized on the surface of uterine luminal and glandular epithelia of pregnant mice 36 h after treatment; the localization was DB3‐specific as it was not seen in mice treated with P3 (using sheep anti‐P3 anti‐idiotype as a probe) or saline. The fluorescent staining reaction in oviduct was weak and only appeared on the surface of the oviducal serosa (peritoneal side). Both DB3 and P3 were also localized in liver (granules W Kupffer cells), kidney (glomerular basement membrane), spleen (on the membrane surface of mononuclear cells in the white pulp), and peritoneal exudate cells (on the membrane surface). Staining could be completely blocked by the addition of the free idiotypes against which the anti‐idiotypes were made but not by the unrelated idiotype. Anti‐idiotypic labelling in vivo is more specific and selective than anti‐whole immunoglobulin labelling.
Distinct Patterns of Virus‐Specific T Cell‐Mediated Cytolysis of Transformed versus Primary Target Cells
Authors: BAENZIGER J. , HENGARTNER H. , ZINKERNAGEL R. M. , GROSCURTH P.
Published: June 2006

Ideas about the mechanism(s) by which cytotoxic T lymphocytes(CTL) lyse appropriate target cells are still controversial. We studied the action of established murine CTL clones as well as of freshly prepared primarily induced CTL against two types of target cell. Transformed tumour cell lines (MC57G and L929) and untransformed cells such as peritoneal exudate cells (PEC) or fibroblasts were examined as target cells by scanning and transmission electron microscopy and with the 51Cr‐release assay. We found independent of which CTL used, that in transformed cells the cell membrane seemed to be the first target of CTL attack, whereas in untransformed cells the first noticeable events appeared to take place in the nucleus of the target cells; the membrane of attacked untransformed PEC or fibroblasts was found to be intact at a time when the cellular organelles already were disintegrated. The morphological observations were paralleled by differences in the kinetics of 51Cr‐release; untransformed target cells released their label only after a 2 h long period, whereas transformed cells released 51Cr considerably earlier.
Synthesis of Soluble C3 and C9 Neoepitopes by Human Alveolar Macrophages in Vitro
Authors: PETTERSEN H. B. , JOHNSON E. , MOLLNES T. E. , GARRED P.
Published: June 2006

The aim of this study was to examine whether soluble neoepitopes of activated C3 (C3b, iC3b, C3c) and C9 are produced by human alveolar macrophages cultured in serum‐free medium. There was a significant and inhibitable production of C3 and C9 neoepitopes and C9 by the macrophages from all donors, as detected by enzyme‐linked immunosorbent assays based on monoclonal (bH6, aE11) and polyclonal (anti‐C9) antibodies. A strong donor‐dependent variation in the levels of the C3 neoepitope and C9 (five‐ to sevenfold) and the C9 neoepitope (twofold) was found. After 1 day (24 h) of incubation, the complement levels were largely unaltered. The presence of an exogenous alternative pathway activator (agarose beads) reduced the amount of soluble complement because of binding to the agarose. However, the relative fraction of C9 neoepitope versus C9 increased (two‐ to threefold), due to agarose mediated activation of C9. The results demonstrate activation of the complement system in serum‐free alveolar macrophage cultures, irrespective of the presence of a known complement activator.
Evidence that Tumour Necrosis Factor (TNF) is not Constitutively Present in Vivo The Association of TNF with Freshly Isolated Monocytes Reflects a Rapid in Vitro Production
Authors: HOFSLI E. , LAMVIK J. , NISSEN‐MEYER J.
Published: June 2006

Tumour necrosis factor (TNF) cytotoxic activity has been shown not to be present in detectable amounts in the serum of healthy humans, but it may be found in some patients with menin‐gococcal disease. In this study we investigated whether TNF is constitutively present in vivo on or within monocytes. TNF was detected on freshly isolated paraformaldehyde‐fixed monocytes from both healthy individuals and cancer patients. No significant difference was found between the two groups. TNF appeared first 40–60 min after in vitro monocyte adherence, which is the same time as it look TNF to appear extracellularly after the exposure of in vitro cultured monocytes to lipopolysaccharide (LPS). This indicates that TNF associated with freshly isolated monocytes was synthesized in vitro. The inducing signal may be monocyte contact with plastic. Exposure of whole blood to LPS immediately after vein puncture was followed by about twice as rapid TNF release as that observed with in vitro cultured monocytes. The release was inhibited by cycloheximide but not by actinomycin D, indicating that the TNF detected did not represent TNF present in vivo. This is consistent with the fact that no TNF cytotoxic activity was detected in blood cell extracts. However, TNF‐mRNA may have been present in vivo. Thus, the available evidence indicates that TNF is not constitutively expressed in vivo.
Polymeric and Monomeric IgA Response in Serum and Milk after Parenteral Cholera and Oral Typhoid Vaccination
Authors: MASCART‐LEMONE F. , CARLSSON B. , JALIL F. , HAHN‐ZORIC M. , DUCHATEAU J. , HANSON L. Å.
Published: June 2006

The effect of vaccinating lactating Pakistani mothers with a combination of live oral typhoid vaccine and parenteral inactivated cholera vaccine on specific milk and serum IgA antibodies in both monomeric (m) and polymeric (p) forms was analysed. IgA antibody titres peaked for both antigenic specificities 2 weeks after the first dose of vaccine. 82±7% of anti‐Vibrio cholerae and 72±17% of anti‐Salmonella typhi IgA were in the polymeric form. These serum pIgA antibodies were mainly dimeric IgA, not complexed with the secretory component. They disappeared more rapidly from serum than mIgA antibodies. Anti‐V. cholerae IgA responses were parallel in serum and milk samples, whereas anti‐S. typhi responses were dissociated. In milk, IgA antibodies were secretory IgA for both antigenic specificities, being probably of local origin in the mammary gland. Our results indicate that both oral and parenteral vaccinations can induce pIgA antibodies in serum and secretions, confirming that the presence of pIgA in serum does not necessarily reflect an immune stimulation only at the mucosal level.
IgG Subclass Deficiency in Patients with Down's Syndrome and Aberrant Hepatitis B Vaccine Response
Authors: AVANZINI M. A. , SÖDERSTRÖM T. , WAHL M. , PLEBANI A. , BURGIO G. R. , HANSON L. Å.
Published: June 2006

Seventeen adult patients with Down's syndrome (DS) and 19 adult healthy references were vaccinated with a hepatitis B vaccine in order to study the IgG subclass response. An enzyme‐linked immunosorbent assay (ELISA) using monoclonal antibodies specific for IgG subclasses was employed. In spite of normal levels of total IgG1 and normal or even high levels of IgG3 in the DS patients, a significantly lower IgG1 response to the vaccine was observed in trisomic patients than in the references.
Effects of Phorbol Esters on B‐Cell Gene Expression
Authors: HöGBOM E. , MÅRTENSSON I.‐L. , FUNA K. , LEANDERSON T.
Published: June 2006

Splenic B lymphocytes were stimulated with lipopolysaccharide alone or in combination with phorbol‐12,13‐dibinyrate, a protein kinasc C‐activating phorbol esier The effect of the treatment was analysed at the single cell level with in situ RNA/RNA hybridization. Hybridization with a kappa light chain probe revealed that the whole population had shifted towards a low, but significant, expression of immunoglobulin mRNA. Analysis at the population level was performed by DNA/RNA and RNA/RNA hybridization experiments. It was found that the steady‐state levels of mRNA for kappa light chain, IgM heavy chain and J chain were reduced by phorbol ester treatment, while the steady‐state level of mRNA for IgD heavy chain was increased. Steady‐state levels of mRNA for Ia antigen and α‐actin were marginally affected.
Regulation of Growth and Differentiating of Pre‐Activated B Lymphocytes
Authors: RAJASEKAR R. , ANDERSSON J. , LEANDERSON T.
Published: June 2006

The differential effects of H‐2 IAk‐specific T helper cells, their soluble factors and Sepharose‐coupled anti‐μ antibodies on the growth and differentiation of pre‐activated B cells were studied. It was found that the prolonged growth of pre‐activated B cells required activation signals from major histocompatibility complex (MHC)‐restricted T helper cells or Sepharose‐coupled anti‐μ antibodies. The MHC‐restricted T helper cells induced both prolonged growth and differentiation of activated B cells Anti‐μ antibodies together with T helper cell‐derived soluble factors induced prolonged growth of activated B cells, but no differentiation into Ig secretion was detected. The inhibition of Ig secretion in anti‐μ cultures could be overcome to some exent by either MHC‐restricted T helper cells or lipopolysaccharide together with soluble factors. It was also observed that T helper cell interactions were needed for long‐term in vitro culture of pre‐activated B lymphocytes.
Production of Gamma Interferon in Mls Disparate Interactions
Authors: GOLDBACH J. , KIRCHNER H. , KÖLSCH E.
Published: June 2006

The marine T‐cell clone E11 isolated from a primary H‐2k histocompatible one‐way mixed lymphocyte culture of B10.BR anti C3H/Tif spleen cells was used to study multidirectional interactions in strong stimulatory Mls disparate responses. Several parameters have recently been studied, and proliferation of T cells upon stimulation by macrophages or B cells of Mls‐disparate stimulator cells, induction of differentiation of B cells of stimulator strains but inhibition of their macrophage differentiation, and the inhibition by E11 T cells of the production of a mitogenic mediator by Mls‐disparate spleen cells have been found. As shown in this paper, these phenomena can be explained by an Mlsa, d, c specific induction of gamma‐interferon(IFN‐γ) production in the responder B10.BR (Mlsb) E11 T cells. It is suggested that (IFN‐γ), as a regulator of feedback mechanisms, plays a critical role in Mls disparate T‐cell stimulation.
Role of T Helper/Inducer Cells as well as Natural Killer Cells in Resistance to Trypanosoma cruzi Infection
Authors: ROTTENBERG M. , CARDONI R. L. , ANDERSSON R. , SEGURA E. L. , ÖRN A.
Published: June 2006

T lymphocytes provide a major line of defence against many protozoan parasites. The aim of this work was to determine the role of T‐cell helper/inducer subset (T h/i) in the resistance to Trypanosoma cruzi in a murine model. The imponatance of natural killer (NK) cells in the resistance to the parasite was also evaluated. BALB/c and C57BL/6 mice were injected with either monoclonal antibodies against L3T4, Thy 1.2, NK1.1, or with a polyclonal rabbit antiserum against NK cells (anti‐asialo GM‐1). The effect of in vivo administration of these antibodies was tested in separate functional assays. Alter antibody treatment, mice were infected with a low dose of T. cruzi in the bloodstream form. Mice depleted of, or reduced in T, T h/i, or NK cell activity all developed higher parasitaemia and had higher mortality than their control counterparts. Mice injected with anti‐L3T4 monoclonal antibodies were unable to generate a specific antibody response to the parasite. Treatment of mice with alpha/beta interferon, which is known to boost NK cell activity, resulted in an enhanced resistance to the parasite. Our data indicate that T h/i cells as well as NK cells are of vital importance in controlling parasitaemia and reducing mortality in T. crazi‐infected mice. Finally, We also demonstrate that the production of antibodies specific for T. cruzi is strictly T helper cell dependent.
Tumour Necrosis Factor‐β Modulates Human Neutrophil‐Mediated Cartilage Damage
Authors: KOWANKO I. C. , BATES E. J. , FERRANTE A.
Published: June 2006

Human neutrophils, when cultured with human articular cartilage coated with heal aggregated immunoglobulin G, degraded proteoglycan and inhibited its synthesis. Neutrophil‐mediated degradation of cartilage was potentiated by recombinant human tumour necrosis factor‐β (TNFβ), although TNFβ alone did not alter proteoglycan degradation. This effect was seen when TNFβ, neutrophils, and cartilage were incubated together, and also when neutrophils were preincubated with TNFβ and washed before being added to cartilage. Similar results were obtained with living and killed cartilage, in contrast, ptetreatment of neutrophils with TNFβ abrogated the neutrophil‐mediated inhibition of proteoglycan biosynthesis. There was no effect of TNFβ alone on synthesis of proteoglycan.
Recognition by Anti‐Fab Antibodies in Rheumatoid Arthritis of Structure(s) Widely Distributed on Human Fab Molecules
Authors: GERARDO S. HUNT , PERSSELIN J. E. , KELD B. , STEVENS R. H.
Published: June 2006

Anti‐Fab antibodies (aFABA) of restricted clonality and acidic spectrotypes were isolated from the sera of patients with rheumatoid arthritis (RA) These aFABA reacted with multiple populations of pooled human Fab molecules, which had been charge separated by chromato‐focusing techniques (CF), indicating that the structures recognized by these aFABA were present on a polyclonal population of Fab molecules. The structures were also widely distributed among the Fab repertoires of normal individuals, as well as individual autologous and heterologous RA patients. Thus, the aFABA did not appear to recognize highly restricted epitope(s), i.e. a private idiotope, limited to its expression to RA individuals. The determinants of the Fab molecules recognized by affinity purified aFABA could be defined in linear and/or conformational structures, depending upon the individual from which the aFABA were isolated. Additionally, some of the affinity purified aFABA also reacted with Fc fragments, suggesting the presence of epibody‐like autoantibodies in this population. Lastly, size analysis of the circulating IgG4 aFABA complexes indicated dial these autoantibodies were not completed with intact IgG, but rather with a molecule of 40–60 kDa, further suggesting the potential for these autoantibodies to react with multiple antigens.
Synergistic Growth‐Inhibitory Activity of Tumour Necrosis Factor and Alpha Interferon Contribution to the Monocyte‐Derived Cytostatic Activity towards Human Leukaemia K562 Cells
Authors: NISSEN‐MEYER J. , AUSTGULEN R. , ESPEVIK T.
Published: June 2006

Monocyte supernatants are cytotoxic towards WEHI 164clone 13 cells and cytostatic towards K562 cells The Cytotoxic towards the clone 13 cells is entirely due to tumour necrosis factor alpha (TNFα), since it was completely neutralized by recombinant TNFα (rTNF) antiserum, and identical dose‐response curves were obtained with supernatants and rTNFα. The Cytostatic activity towards the K562 cells, however, was only partly due to TNFα, since this activity was only partly neutralized by the rTNFα antiserum. Moreover, the supernatants were found to be more cytostatic towards K562 cells than rTNFα preparations that contained comparable amounts of TNF. Alpha interferon (IFN‐α also contributed to cytostasis of the K562 cells, since antibodies against IFN‐α partially inhibited the cytostatic activity in the supernatants, and further inhibition, although not complete, was obtained in the presence of both IFN‐α antibodies and rTNFα antiserum Moreover, recombinant IFN‐α‐2c (rIFN‐α‐2c) inhibited the growth of K562 cells, acting synergistically with rTNFα. Upon cation exchange chromatography, natural TNF in the monocyte supernatant eluted more broadly than rTNF, and significant amounts of TNFα were found in all column fractions containing cytostatic activity.
Susceptibility to Tolerance Induction of Bursal and Peripheral B Cells
Authors: VAINIO O. , TOIVANEN P. , GRANFORS K. , PINK J. R. L.
Published: June 2006

We investigated cellular aspects of immunological tolerance to protein antigens in chickens by examining the immune responses of bursal and splenic cells from tolerant or normal chickens after transfer into cyclophosphamide (CP)‐treated recipients. Newly‐hatched chicks were made tolerant to bovine serum albumin (BSA) by injection of 100 mg of the antigen. When bursa cells from 4‐day‐old BSA‐unresponsive chicks were transferred into CP‐treated recipients, the reconstituted birds were able to respond to a subsequent injection of BSA almost as well as normal birds, and as well as CP‐treated birds that had been reconstituted with normal bursa cells. To investigate whether the presence of the BSA antigen might affect recovery from tolerance, we injected CP‐treated recipients with BSA at the time of transfer of bursal cells. The presence of the antigen prevented the recovery of the anti‐BSA response in reconstituted birds. When spleen cells from 6.5‐week‐old unresponsive chicks were transferred into CP‐treated recipients, no recovery of responsiveness to BSA could be demonstrated. A likely reason for the failure of splenic B cells to recover responsiveness on transfer is their inability to generate somatic variants of Ig genes in the same way as bursal stem cells. Thus, when the bursa involutes, the chicken's antibody repertoire may be frozen in a less adaptable state than that of a mammal.
Mitochondrial Autoantigens in Primary Biliary
Authors: FROSTELL Å. , MENDEL‐HARTVIG I. , NELSON B.D. , TÖTTERMAN T.H. , BJÖRKLAND A. , RAGAN I.C. , CLEETER M.W.J. , PATEL S.D.
Published: June 2006

Anti‐mitochondrial autoantibodies (AMA) from patients with primary biliary cirrhosis (PBC) were analysed for fine specificity by immunoblotting and enzyme‐linked immunosorbent assay (ELISA). Inhibition ELISA showed that complex I (NADH‐ubiquinone reductase) from beef heart mitochondria completely inhibited the binding of AMA to mitochondrial inner membranes (SMP), indicating that the major mitochondrial antigens are located in complex I, Immunoblot analysis of beef heart SMP, complex I and the iron sulphur (IP) subfraction of Complex I revealed several antigens, one of which (75 kDa) reacted with all PBC sera but not with the additional autoimmune sera tested. Resolution of SMP or complex I by two‐dimensional electrophoresis yielded in both preparations a polypeptide of 75 kDa with an isoelectric point of 6.4, which reacted with PBC serum and with rabbit antisera raised against the 75,000 subunit of complex I. In immunoblot experiments, the antigenicity of the 75,000 polypeptide in SMP, complex I, and the IP subfraction is increased by prior reduction of the sample with mercaptoethanol. This suggests a similarity to the PBC‐specific ‘M‐2’antigen, which is also sensitive to sulphur reagents. The data indicate that the 75 kDa polypeptide of complex I is a major mitochondrial antigen binding AMA in PBC sera, and allows us to identify the location and probable function of the PBC antigen.
Spontaneous Lymphocyte Proliferation during Trauma and Infection
Authors: SOOP M. , SOOP A. , SUNDQVIST K.‐G.
Published: June 2006

Spontaneous lymphocyte proliferation (SpP), measured in vitro as the rate of [14C]thymidine incorporation in blood lymphocytes, was investigated in non‐infected postoperative patients, infected postoperative patients, and healthy volunteers, with 72, 24, and 3 h of lymphocyte culture. With 24‐h cultures, infected postoperative patients showed 17‐fold higher SpP than non‐infected postoperative patients (2527±1552 versus 151±77 cpm, mean± SD, P<0.001) and 37‐fold higher SpP than healthy volunteers (P<0.001). Postoperative patients without infection had twice as high SpP as healthy volunteers (P<0.001). Lymphocytes harvested after 24 h of cell culture showed significantly higher SpP than corresponding values at 72 and 3 h, in patients as well as in healthy volunteers (P<0.01). Infected postoperative patients showed a higher SpP than non‐infected patients after only 3 h of cell culture (270±192 versus 48±10 cpm, P<0.001). An inverse correlation was observed between the level of SpP and body temperature in patients with postoperative infection (r=‐0.62, P<0.05). The results indicate that lymphocytes are activated by uncomplicated surgery and particularly by postoperative infection, and that characteristics of SpP are reproducible in short cell‐culture periods, which suggests that in vitro measurements of SpP may be of value in the detection of severe postoperative infection.
Inhibition of Neutrophil Chemotaxis by a Monoclonal Antibody (TM316)
Authors: SHIMIZU A. , TAKEUCHI A. , OHTO H. , HASHIMOTO T. , MIYAMOTO T.
Published: June 2006

A monoclonal antibody, TM316, IgM, was raised against the human monocytoid leukaemia cell line THP‐1, and was shown to inhibit polymorphonuclear leucocyte (PMN) chemotaxis. The molecular weight (MW) of the protein on the PMN membrane with which TM316 bound was about 78,000. TM316 inhibited the chemotactic response of human PMN induced by at least three kinds of chemotactic factors (activated serum, N‐formyl‐l‐methionyl‐l‐leucyl‐l‐phenylalanine (FMLP), and a lymphocyte‐derived chemotactic factor (LDCF)) to the same extent. The extent of inhibition of chemotaxis by TM316 was strongly correlated with the quantity of cellular surface antigen recognized by TM316, when a cell sorter was used for analysis. TM316 did not alter the number of Fc or complement receptors of PMN, nor did it affect luminol‐enhanced chemiluminescence(CL), lysosomal enzyme release, adherence, or superoxide anion generation by PMN. TM316 seemed to recognize a common surface antigen which was necessary only for the process of chemotaxis.
Inhibitory Effects of Histamine on Interleukin 2 and Gamma Interferon Production by Different Human T Helper Cell Subsets
Authors: DOHLSTEN M. , HEDLUND G. , SJÖGREN H.‐O. , CARLSSON R.
Published: June 2006

We have previously demonstrated that histamine can inhibit human helper T cells by direct interaction with these cells. It has now been investigated whether histamine inhibits lymphokine production by various subsets of CD4+ human T cells separated with the Leu‐8 (p80) and Leu‐18 (anti‐CD45R; p220) monoclonal antibodies (MoAb). Histamine was shown to suppress to a similar extent the production of interleukin 2 (IL‐2) and gamma interferon (IFN‐γ) by Leu 3+, Leu 3+8+, and Leu 3+8− cell subsets. Mitogen‐activated, unseparated Leu 3+ and purified Leu 3+8− cells produced maximal amounts of IL‐2 after 24 h and IFN‐γ after 72 h of culture. In contrast, the Leu 3+18+ subset produced no IL‐2 after 24 h, and maximal amounts of IL‐2 no sooner than 48 h of culture, and only small amounts of IFN‐γ during the entire culture period of 96 h. Histamine suppressed the production of IL‐2 by both subsets, both when produced early (after 24 h), as in the case of the Leu 3+18− subset, and late (after 48 h of culture), as for the Leu 3+18+subset. The IFN‐γ production by the Leu 3+ and Leu 3+18−cells and the marginal production by Leu 3+18+ cells were significantly suppressed by histamine. Dual staining with Leu 8 and Leu 18 MoAb demonstrated that the Leu 18− cell compartment included both Leu 8+ and Leu 8− cells. It was shown that the inhibitory effect of histamine on the early production of IL‐2 and the major production of IFN‐γ by T helper cells is mediated via action on both the Leu 3+18−8− and the Leu 3+18−8+ cells. The inhibitory effect of histamine on the late production of IL‐2 is mediated mainly via action on Leu‐18+ cells.
An Altered Response by Psoriatic Keratinocytes to Gamma Interferon
Authors: BAKER B. S. , POWLES A. V. , VALDIMARSSON H. , FRY L.
Published: June 2006

To determine whether psoriatic keratinocytes differ from normal keratinocytes in their response to gamma interferon, epidermal cell suspensions from normal and from lesional and uninvolved psoriatic skin were cultured in the presence of gamma interferon and the induction of HLA‐DR expression and inhibition of cell growth were measured The addition of 102 units of gamma interferon/ml during 7‐day culture period significantly increased mean HLA‐DR+ cell numbers in 21 epidermal suspensions of normal from 3.9 to 24.1% (P<0.0001), uninvolved psoriatic from 8.4 to 33.1% (P<0.01), and to a lesser extent lesional psoriatic biopsies from 12.6 to 18.3% (P<0.01). However, the increase in HLA‐DR+ cell numbers in these latter cultures was significantly less than that observed in either normal or uninvolved psoriatic epidermal cell cultures (P<0.0001). Furthermore, [3H]thymidine incorporation was substantially decreased by gamma interferon in 16 out of 22 (73%) cultures of normal epidermal cells; this decrease was statistically significant (P<0.0l). In contrast, only 4 out of 11 (36%) lesional and 9 out of 21 (43%) uninvolved psoriatic epidermal cultures showed comparable inhibition of proliferation. These findings suggest that psoriatic keratinocytes have an altered response to gamma interferon: this could explain the infrequency of keratinocyte HLA‐DR expression in psoriatic plaques in vivo and may also contribute to the increased epidermal proliferation that characterizes this disease.
Monoclonal Antibodies against Leucoagglutinin‐Reactive Human T‐Lymphocyte Surface Components
Authors: HAMMARSTRÖM M.‐L. , BERZINS T. , AGUILAR‐SANTELISES M. , ANDERSSON G. , PERLMANN P. , HAMMARSTRÖM S.
Published: June 2006

The binding specificities of three biologically active anti‐lymphocyte monoclonal antibodies (MoAb) (K46M, K3G, and 3‐19‐2) produced against human T‐cell surface components reactive with the mitogenic lectin leucoagglutinin from Phaseolus vulgaris (La) were analysed. K46M is a strong T‐cell mitogen, while K3G and 3‐19‐2 inhibited cell‐mediated cytotoxicity. Resting peripheral blood lymphocytes (PBL) contained 4‐16% K46M+ cells, 8‐35% K3G+ Cells, and <0.3–4% 3‐19‐2+ cells. After stimulation with T‐cell mitogens the proportion of K46M+ and 3‐19‐2+ cells increased markedly (mean 59 and 30% positive cells, respectively), while the increase in K3G+ cells was less prominent (38%). K46M‐reactive structures were expressed on mature T cells and probably also on B cells. K3G reacted with B and T cells while 3‐19‐2 showed a broader specificity reacting also with erythrocytes. All three MoAb reacted with lipid extracts of resting and activated PBL as well as with purified neutral glycolipids of lymphoid origin. In addition 3‐19‐2 reacted with lipid extracts of erythrocytes. K‐46M immunoprecipitated four surface peptides from lectin‐stimulated PBL. Their apparent molecular weights were 53,000, 42,000, and 16,000(doublet). The 53,000 and 42,000 MW peptides were identified as the α and β chains of the T‐cell antigen receptor. The identity of the 16,000 MW peptides is presently unknown. K3G and 3‐19‐2 did not specifically precipitate any lymphocyte surface peptide.