bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of root‐ and stolon‐infecting fungi on root‐colonizing nematodes of white clover
Authors: Zahid M. I. , Gurr G. M. , Nikandrow A. , Hodda M. , Fulkerson W. J. , Nicol H. I.
Published: March 2003

The effects on white clover (Trifolium repens) of different combinations of the nematodes Meloidogyne trifoliophila, Helicotylenchus dihystera and Heterodera trifolii and nine stolon‐infecting and three root‐infecting fungi were studied in a glasshouse experiment. The presence of the fungus Phytophthora megasperma alone increased (P < 0·001) root‐rot severity and reduced (P < 0·001) plant growth. Other species combinations, such as Phoma nebulosa and Alternaria alternata, interacted and increased root‐rot severity. Combinations of P. megasperma with Pythium irregulare, and P. nebulosa with Phoma medicaginis or A. alternata, increased M. trifoliophila populations. Several other fungi (P. irregulare, P. nebulosa, Colletotrichum coccodes, Macrophomina phaseolina, P. medicaginis and Phoma sp.) interacted with the nematode M. trifoliophila causing severe root‐knot symptoms. The results indicated that fungi and nematodes interacted to cause root and stolon rot and reduced yields, and that poor persistence of white clover in pastures is likely to be a problem with a complex etiology.
Botryosphaeria canker of Cistus ladanifer
Authors: Sánchez‐Hernández M. E. , Gutiérrez‐García J. , Trapero‐Casas A.
Published: March 2003

The shrub Cistus ladanifer (gum cistus, rock rose) is a key component of the Maquis in the Mediterranean forests of southern Spain and Portugal. Between 1996 and 1998, and coinciding with exceptionally high rainfall rates, a disease characterized by foliar wilting and death of branches, leading to death of the whole plant, affected C. ladanifer in the Mediterranean oak and pine forests in Andalucía (southern Spain). Field surveys carried out in 15 forest areas showed that diseased branches were associated with the presence of strips of necrotic bark. Botryosphaeria dothidea was consistently obtained from the necrotic tissues. Growth of isolates on potato dextrose agar was very rapid at high temperatures, with a growth rate of 25 mm day−1 at the optimum temperature (about 28°C). Germination rate of conidia increased with temperature, with the optimum temperature being over 35°C. Pathogenicity tests were conducted on cut branches, 3‐year‐old seedlings and mature plants in the field. In every situation, disease symptoms were reproduced in the inoculated branches and Koch’s postulates were fulfilled. The results demonstrate the behaviour of B. dothidea as a primary pathogen of C. ladanifer. The present work constitutes the first description of fungal canker of C. ladanifer caused by B. dothidea, being the first report of a disease affecting this shrub species.
Factors affecting symptom production by latent Botrytis cinerea in Primula×polyantha
Authors: Barnes S. E. , Shaw M. W.
Published: March 2003

Latent infection by Botrytis cinerea was frequently detected in young Primula×polyantha (horticultural hybrid polyanthus) plants. Genetically marked isolates were used to demonstrate that conidial inocula applied to young plants generally did not result in disease appearing on the leaves until plants flowered, regardless of when plants were inoculated. The severity of disease was proportional to the length of each day spent at 100% relative humidity during the latent period, regardless of time or form of inoculum. Plants with a prior B. cinerea infection were more susceptible to further infection and also showed symptoms of B. cinerea infection more quickly. Bacteria found to be associated with B. cinerea increased the likelihood of B. cinerea symptoms being seen on the plant, but did not directly assist infection.
Evaluation of potential biocontrol agents against Claviceps africana in vitro and in vivo
Authors: Bhuiyan S. A. , Ryley M. J. , Galea V. J. , Tay D.
Published: March 2003

Undiluted culture filtrates of two commercial products of Trichoderma spp., Trichopel and Trichoflow, and two isolates of Penicillium citrinum completely inhibited the conidial germination of macroconidia of Claviceps africana, the cause of ergot or sugary disease of sorghum (Sorghum bicolor) in vitro. Similarly, Pseudomonas aeruginosa and Burkholderia cepacia completely inhibited macroconidial germination, with the former being more effective at high dilutions. In contrast, these bacterial isolates failed to inhibit infection in vivo in glasshouse tests with ergot‐inoculated sorghum, but all fungal biocontrol agents (including an isolate of Epicoccum nigrum) reduced the severity of disease (percentage of infected spikelets per panicle), in some cases completely inhibiting the development of ergot. In a second glasshouse trial, optimum control was achieved when the biocontrol agents were applied 3–7 days before inoculation with conidia of C. africana.
Characterization of a New IgE‐Binding 35‐kDa Protein from Birch Pollen with Cross‐Reacting Homologues in Various Plant Foods
Authors: Vieths  , Frank  , Scheurer  , Meyer  , Hrazdina  , Haustein
Published: March 2003

The present investigation was undertaken to obtain molecular data of a new immunoglobulin (Ig)E‐binding birch pollen protein with a mass of 35 kDa. In a previous study, this protein showed IgE cross‐reactivity with 34‐ and 35‐kDa proteins in apples, pears, carrots, bananas and other exotic fruits. Since the protein was N‐terminally blocked, it was purified by preparative SDS‐PAGE, and multiple proteolytic fragments were subsequently generated by in‐gel digestion with the endoproteinases Glu C, Lys C and Clostripain. After electrophoretic separation and blotting onto polyvinylidene difluoride (PVDF), the resulting polypeptides were subjected to N‐terminal amino acid microsequencing. The internal sequences obtained showed a high degree of sequence identity to isoflavone reductases (IFR) and isoflavone reductase‐like proteins (IRL) from several plants which also had a similar size. For a stretch of 25 consecutive residues this identity ranged from 56% for IFR from peas and chick peas and an IRL from maize, to 80% for a tobacco IRL. A 453 bp fragment was amplified from total birch pollen RNA by polymerase chain reaction (PCR) using primers derived from the nucleotide sequence of the tobacco IRL. The deduced 151 amino acid sequence represented ≈ 50% of the protein and confirmed the sequence identities obtained by Edman degradation. Moreover, the 25 amino acid sequence was included in the cloned fragment. Deduced and determined amino acids showed only one mismatch, which was due to a single nucleotide exchange. At the antibody level, the immunological relationship of the birch pollen protein to IRL and IFR was demonstrated by immunoblotting with a rabbit antiserum against a pea IFR which recognized the same birch protein as patients' IgE. The rabbit antiserum also reproduced the cross‐reactivity pattern previously observed with patients' IgE by recognizing related proteins in specific plant foods, including some exotic fruits. We therefore suggest that the 35‐kDa birch pollen protein belongs to the IFR/IRL family and represents a minor allergen, possibly being responsible for less common pollen‐related food allergies in patients allergic to birch pollen.
Presence of the IL‐1RA Allele 2 (IL1RN*2) is Associated with Enhanced IL‐1β Production In Vitro
Authors: Santtila  , Savinainen  , Hurme
Published: March 2003

The genes of the interleukin‐1 (IL‐1) complex code for three proteins: IL‐1α, IL‐1β and the IL‐1 receptor antagonist (IL‐1RA). Each of these genes is polymorphic and there is increasing evidence that certain alleles are associated with increased susceptibility to a given disease of inflammatory nature. In the IL‐1β gene there are two base‐exchange polymorphisms in positions −511 and +3953, and IL‐1RA gene has a penta‐allelic polymorphic site in intron 2 containing variable numbers of an 86‐bp tandem repeat sequence. As the IL‐1β/IL‐1RA ratio may be critical in the regulation of inflammation, we examined whether there are allelic associations between these loci (thus suggesting co‐ordinate regulation) and whether these have an effect on the in vitro production of IL‐1β. We found that the IL‐1RA allele 2 (IL1RN*2) is associated with the presence of allele 2 of the IL‐1β gene (position −511) and with the absence of allele 2 of the IL‐1β gene (position +3953). Mononuclear cells from carriers of allele 2 (position −511) and non‐carriers of allele 2 (position +3953) had a slight, but non‐significant, elevated capacity to produce IL‐1βin vitro. However, IL‐1RA allele 2 strongly increased in vitro production of IL‐1β, regardless of the presence or absence of these alleles. Taken together, these data suggest that the known allelisms in the IL‐1β gene are not major regulators of the in vitro IL‐1β production, but the IL‐1RA allele 2 (or an unknown allele strongly associated with it) has a decisive role.
A Glycosylated Bence Jones Protein and its Autologous Amyloid Light Chain Containing Potentially Amyloidogenic Residues
Authors: Foss  , Nilsen  , Cornwell  , Husby  , Sletten
Published: March 2003

Amyloidosis is characterized by deposition of protein fibrils in various tissues. The wide variety of sequences of both amyloidogenic and non‐amyloidogenic immunoglobulin light chains makes them a unique tool for addressing the importance of primary structure in the formation of insoluble fibrils. In this study, we have determined the primary structure of the κ I immunoglobulin light chain from both the urinary Bence Jones protein and the deposited amyloid fibrils of a patient (MH) with primary amyloidosis. The sequence identity of urinary‐excreted and tissue‐deposited light chains excluded biclonality and somatic mutations and confirmed that the light chain existed in both a soluble and an insoluble form. Several residues have been previously reported to be significantly associated with amyloidogenic κ chains. Many of these were found in the MH sequence, including Asp31, Asn45, Phe49, Gln55, His70, Asn/Gly93 and Pro/Val96, thereby supporting their potential role in fibrillogenesis. In addition, Asn20 and Pro60 of protein MH replaced the normally conserved Thr20 and Ser60. Asn20 was glycosylated in both the Bence Jones and the amyloid fibril protein MH. Cumulative effects of amyloid‐associated residues and glycosylation might have rendered the immunoglobulin light chain MH prone to fibril formation.
Reconstitution of Opsonizing Activity by Infusion of Mannan‐Binding Lectin (MBL) to MBL‐Deficient Humans
Authors: Valdimarsson  , Stefansson  , Vikingsdottir  , Arason  , Koch  , Thiel  , Jensenius
Published: March 2003

Mannan‐binding lectin (MBL, previously named mannan‐binding protein, MBP) is a serum collectin, which activates complement upon binding to microbial carbohydrates. This results in opsonization of the micro‐organisms as well as direct complement‐mediated killing. Clinical evidence indicates that MBL has an important role in the innate immune defence against various pathogens. Genetically determined MBL deficiency is associated with increased susceptibility to infections. We have infused two MBL‐deficient individuals with clinical grade MBL, purified from pooled donor plasma, in doses sufficient to attain normal concentration of MBL in serum. This resulted in normalization of complement‐mediated opsonization. An initial rapid decrease in the serum concentration of MBL was followed by a slower decline with an estimated half‐life of about 6 days. No adverse effects were observed and anti‐MBL antibodies could not be detected following several MBL infusions. One of the two MBL recipient, a two‐year‐old girl, who had been suffering from repeated infections from the age of 4 months, was given a total of six MBL infusions. She has subsequently remained healthy for more than three years.
Polymorphism of the IL‐1 Gene Complex in Epstein–Barr Virus Seronegative and Seropositive Adult Blood Donors
Authors: Hurme  , Helminen
Published: March 2003

Epstein–Barr virus (EBV) seronegativity is rare in adults. To examine whether genetic differences would explain this, we studied the genetic polymorphisms of the genes of the interleukin‐1 (IL‐1) complex in seronegative adults. These cytokines (i.e. IL‐1α, IL‐1β and IL‐1 receptor antagonist, IL‐1RA) regulate, in several ways, the inflammatory reactions of the body. In each of these genes there are polymorphic sites and the various alleles differ in their frequency in several diseases of inflammatory nature. In 400 healthy blood donors (from 18 to 60 years of age) there were 20 (5%) seronegative persons. The frequency of allele 2 of the IL‐1β gene (base exchange polymorphism at position −511 from the transcriptional start site) was decreased in the seronegative patients (0.20 versus 0.42 in the seropositive patients, P < 0.05, χ2‐test). Moreover, the frequency of allele 2 of the IL‐1RA (polymorphism defined by variable numbers of 86‐bp repeats in intron 2) was slightly, but not significantly, decreased in the seronegative patients. Alleles of these two loci are known to be associated, but in the seronegative patients this association was abnormal: 11 out of 20 (55%) were of the IL‐1RA‐2 negative/IL‐1β‐2 negative type, while of the seropositive patients, 25% were of this type (P < 0.01, χ2‐test). These data suggest that immunological differences, depending on cytokine gene polymorphisms, regulate the resistance to EBV infection.
Comparison of the Frequencies of Arginines in Heavy Chain CDR3 of Antibodies Expressed in the Primary B‐Cell Repertoires of Autoimmune‐Prone and Normal Mice
Authors: Krishnan  , Marion
Published: March 2003

Because the pathogenesis of anti‐DNA Ab in SLE is correlated to Ab specificity for native DNA (dsDNA), understanding how such specificity is generated is important. The VH structures of most autoimmune anti‐DNA antibodies include at least one arginine in VH–CDR3; moreover, antibody specificity for dsDNA can be correlated to the relative position of arginines in VH–CDR3. The coding sequences for most VH–CDR3 arginines among the anti‐DNA MoAbs we have studied to date appeared to have been encoded by sequences generated during V–D–J recombination and would have been expressed in the primary B‐cell repertoire. The frequency at which arginine codons are generated during V–D–J recombination therefore could potentially influence the frequency at which DNA‐specific B cells are generated in the primary B‐cell repertoire. The present study was undertaken to determine whether a higher percentage of B cells in the primary, preautoimmune repertoire of autoimmune‐prone (NZB × NZW)F1 mice have immunoglobulin heavy chains with at least one VH–CDR3 arginine compared to B cells in the primary, preimmune repertoire of non‐autoimmune‐prone BALB/c mice. The present results indicate that mature B cells in preautoimmune (NZB × NZW)F1 mice, whether specific for DNA or not, are no more likely to have heavy chains with VH–CDR3 arginines than are B cells in BALB/c mice. The high frequency of recurrence of VH–CDR3 arginines among autoimmune anti‐DNA in (NZB × NZW)F1 mice would appear to derive from the selective oligoclonal expansion of selected B cells that express such structures.
Beneficial Role of Human Endogenous Retroviruses: Facts and Hypotheses
Authors: Larsson  , Andersson
Published: March 2003

Human endogenous retroviruses (HERVs) have recently been suggested as mediators of normal biological processes such as cellular differentiation and regulation of gene expression. Moreover, a direct role for HERVs in pathogenesis and the development of disease is now better appreciated. Elucidation of the mechanisms regulating HERV biology should provide information about fundamental cellular activities and the pathogenesis of multifactorial diseases such as cancer and autoimmune disease. The importance of understanding the roles of HERVs is underscored by the recently obtained insight that activation of endogenous retroviruses poses potential risks following xenotransplantation and in gene therapy using retroviral vectors. Furthermore, HERV‐encoded superantigens have recently been implicated as causes of autoimmune disease. This review discusses the established and possible biological roles of HERVs, and proposes hypotheses concerning their involvement as mediators of fundamental cellular responses. We propose that the evolutionary persistence of endogenous retroviruses in the genomes of eukaryotic cells reflects their indispensability in important normal functions in specialized cellular environments. HERVs can also be potentially hazardous through their involvement in the development of disease. In addition, the creation of new retroviruses can occur through recombination, between different HERVs and between HERVs and exogenous retroviruses.
Pathophysiological Roles of Calreticulin in Autoimmune Disease
Authors: Eggleton  , Llewellyn
Published: March 2003

Autoantibodies against the endoplasmic reticulum (ER) luminal protein, calreticulin are often present in sera from patients with systemic lupus erythematosus, rheumatic disease and various parasitic diseases including onchocerciasis. New information has revealed that calreticulin is implicated in a number of autoimmune processes, including molecular mimicry, epitope spreading, complement inactivation and stimulation of inflammatory mediators, such as nitric oxide production. Calreticulin also binds to the Ro/SS‐A antigen complex, which is composed of at least three immunologically distinct proteins bound to a group of small cytoplasmic RNAs that together form a common target for autoimmune responses. Up‐regulation of calreticulin at the protein and RNA levels can be triggered by cell stresses, including heat shock, exposure to heavy metals and perturbation of normal ER function, which may in some cases lead to its secretion from cells. Calreticulin is targeted by autoantibodies following its release into the extracellular environment, possibly as a result of cell death, or its presence at the cell surface in response to insults such as viral infection or ultraviolet irradiation. These findings suggest that calreticulin is not just an autoantigen, but plays an active role in the pathology of various autoimmune disease through determinant spreading.
A Mouse Model for Slowly Progressive Primary Tuberculosis
Authors: Mustafa  , Phyu  , Nilsen  , Jonsson  , Bjune
Published: March 2003

The progression from primary Mycobacterium tuberculosis infection to disease is usually slow in humans. The aim of this study was to develop and characterize a mouse model for slowly progressive primary tuberculosis, using the intraperitoneal (i.p.) route of infection, and to compare it with our previously described model of latent M. tuberculosis infection. B6D2F1 hybrid mice inoculated with 1.5 × 106 colony‐forming units (CFUs) of M. tuberculosis H37Rv were followed‐up for 70 weeks. Lungs, livers and spleens were examined for bacillary growth, histopathological changes and mycobacterial antigens (MPT64, ManLAM and multiple antigens of M. tuberculosis), by immunohistochemical staining. The infection was found to pass through three distinctive phases. During phase 1, mice were healthy despite development of small granulomas and an increasing number of bacilli in the lungs. During phase 2, mice were unwell but mortality was low. The count of M. tuberculosis and the granuloma size stabilized. The granulomas contained an increasing population of large, vacuolated macrophages. During phase 3, mice became moribund and died rapidly, but the M. tuberculosis count remained relatively stable. The inflammatory infiltrates filled ≈ 80% of the lung parenchyma and the lesions were not well demarcated. Rapidly progressing inflammation, rather than an increase in the M. tuberculosis count, seems to contribute more to mortality.
Monocyte Rescue of Human T Cells from Apoptosis is CD40/CD154 Dependent
Authors: Sakata  , Sakata  , Kong  , Vela‐Roch  , Talal  , Dang
Published: March 2003

The induction of T‐cell apoptosis is regulated in part by monocytes (CD14+ cells). Human peripheral blood monocytes inhibited the spontaneous cell death of activated T cells in vitro. The inhibition of T‐cell apoptosis did not require autologous monocytes. Inhibition required direct contact with monocytes and was not due to a soluble factor. Furthermore, treatment of monocytes with actinomycin D, cycloheximide and paraformaldehyde abrogated the anti‐apoptotic activity of these cells. Blocking antibody to CD40 and CD154 (CD40 ligand) decreased the ability of monocytes to aid in T‐cell survival, whereas, blocking LFA‐1/I‐CAM‐1, Fas ligand and the CD4/major histocompatibility complex class II interaction did not affect the influence of monocytes on T‐cell survival. This shows that monocytes rescue of activated T cells from apoptosis is dependent upon CD40/CD154 interaction.
In Vitro and In Vivo Bioactivity of Single‐Chain Interleukin‐12
Authors: Foss  , Moody  , Murphy Jr  , Pazmany  , Zilliox  , Murtaugh
Published: March 2003

Interleukin‐12 is a heterodimeric cytokine with potent immunoregulatory properties, making it a potential vaccine adjuvant and an immune response modulator. The study of its function is confounded by its heterodimeric structure. In order to facilitate the study of interleukin‐12 in both in vitro and in vivo models, we constructed a single‐chain porcine interleukin‐12 gene and expressed the recombinant protein in Pichia pastoris. Single‐chain porcine interleukin‐12 was bioactive in vitro on both human and porcine cells as measured by its ability to induce proliferation of lymphoblasts and interferon‐γ secretion by lymph node cells. In contrast, the p40 subunit of porcine interleukin‐12 alone did not induce proliferation or inhibit the activity of the single‐chain porcine interkeukin‐12. The in vivo bioactivity of single‐chain porcine interleukin‐12 was demonstrated in an oral immunization model where it increased antigen‐specific IgA and IgG in jejunal mucus. These results indicate that binding of interleukin‐12 to its receptor and transduction of intracellular signals requires both p40 and p35 subunits. The bioactivity of interleukin‐12 expressed as a single polypeptide will facilitate its in vivo delivery and study of its structure and function.
Expression of Monocyte Chemoattractant Protein‐1 in Kawasaki Disease: The Anti‐Inflammatory Effect of Gamma Globulin Therapy
Authors: Asano  , Ogawa
Published: March 2003

We investigated the expression of monocyte chemoattractant protein‐1 (MCP‐1) mRNA, protein, and its bioactivity in plasma, mononuclear cells (MNC) and polymorphonuclear leukocytes (PML) in patients with Kawasaki disease, including those who were treated with intravenous gamma globulin. MCP‐1 mRNA expression was increased in the MNC and the plasma level of MCP‐1 and monocyte chemotactic activity in plasma in the acute phase as compared with healthy control levels and decreased after the gamma globulin therapy. The infused gamma globulin contained MCP‐1 protein with monocyte chemotactic activity and did not show a neutralising effect against MCP‐1 protein in vitro. Our results suggest that the infusion of gamma globulin may reduce the production of MCP‐1 in MNC in patients with Kawasaki disease, subsequently reducing its level in plasma. The changes in MCP‐1 level after gamma globulin therapy may serve to alleviate the symptoms in the acute phase in patients with Kawasaki disease.
Enhancement of Experimental Autoimmune Encephalomyelitis Severity by Ultrasound Emulsification of Antigen/Adjuvant in Distinct Strains of Mice
Authors: Määttä  , Nygårdas  , Hinkkanen
Published: March 2003

Susceptibility to experimental autoimmune encephalomyelitis (EAE) is associated with the major histocompatibility complex (MHC) haplotype. In this study EAE could be induced in six out of ten mice of the resistant DBA/2 (H‐2d) strain by ultrasound emulsified antigen/adjuvant, whereas none of the mice immunized with the conventional adjuvant developed the disease. Similar results were previously obtained for the MHC identical BALB/c mice. Further, while only few T cells were present in the central nervous systems (CNS) of the diseased DBA/2 mice, macrophages formed the majority of the infiltrates. In congenic BALB.B (H‐2b) and BALB.K (H‐2 k) mice, EAE could be induced with both sonicated and extruded antigen/adjuvant emulsion. The results indicate that the EAE resistance in mice carrying the H‐2d MHC haplotype is dependent on the physical structure of the immunogen.
Binding of Immune Complexes to Erythrocyte CR1 (CD35): Difference in Requirement of Classical Pathway Components and Indication of Alternative Pathway‐Mediated Binding in C2‐Deficiency
Authors: Klint  , Gullstrand  , Sturfelt  , Truedsson
Published: March 2003

Deficiency of complement components within the classical pathway is associated with increased risk for immune complex disease. However, C2‐deficient individuals often have a mild disease and about 50% are healthy. To study the importance of the different components for immune complex clearance, bovine serum albumin (BSA)/anti‐BSA complexes were opsonized in human serum and the binding to erythrocyte complement receptor type 1 (CR1, CD35) was measured in vitro. In C2‐depleted serum the complexes were opsonized and bound to CR1 but the reaction needed a longer opsonization time than in normal human serum (NHS). In contrast, serum reagent lacking C1q, C4 or C3 did not promote binding in this assay system. We also demonstrated that elevated levels of factor B could restore binding of complexes to erythrocytes in C2‐depleted serum via alternative pathway activation. These results indicate that in spite of lack of a complete classical pathway, C2‐deficient individuals could retain some immune complex opsonizing activity via the alternative pathway. This finding could contribute to the understanding of differences in association between complement deficiency and immune‐complex disease.
Dexamethasone, but not IL‐1 Alone, Upregulates Acute‐Phase Serum Amyloid A Gene Expression and Production by Cultured Human Aortic Smooth Muscle Cells*
Authors: Kumon Y. , Suehiro T. , Hashimoto K. , Sipe J. D.
Published: March 2003

Although the SAA1 and SAA2 protein isoforms (A‐SAA) of the serum amyloid A (SAA) family of acute phase reactants have been found in a number of extrahepatic tissues; the site of synthesis of extrahepatic SAA remains to be clarified. To investigate site(s) of synthesis of the SAA protein localized to atherosclerotic plaque, expression of the SAA1 and SAA2 genes by cultured human aortic smooth muscle cells (HASMC) was investigated. A‐SAA protein isoforms were detectable by immunoblot analysis in the culture medium of HASMC. Both A‐SAA and C‐SAA (SAA4) mRNA isoforms were constitutively expressed by HASMC, but not, however, by the human umbilical vein endothelial cells. Expression of A‐SAA mRNA by HASMC was upregulated by corticoid hormones including dexamethasone (Dex), corticosterone, hydrocortisone, and aldosterone, but not by the cytokines interleukin (IL)‐1, IL‐6, and tumour necrosis factor (TNF)‐α alone. Dex stimulation of A‐SAA mRNA was time and dose dependent from 6 to 48 h. The threshold concentration for upregulation of A‐SAA mRNA in HASMC by Dex was between 0.1 and 1 nm. IL‐1, known to upregulate extrahepatic A‐SAA gene expression in other cell systems only slightly, if at all, upregulated Dex‐induced A‐SAA expression by HASMC. Thus, it is possible that some of the A‐SAA protein in the vascular wall (atherosclerotic plaques) can originate from smooth muscle cells. In consideration of recent reports that A‐SAA modulates the inflammatory process and lipid synthesis, A‐SAA can potentially serve as a physiological regulator of smooth muscle cell homeostasis within that, in a disease state, participates in the formation of atherosclerotic plaques.
Role of IgE in Primary Murine Schistosomiasis mansoni
Authors: El Ridi R. , Ragab S. , Lewis S. , Afifi A.
Published: March 2003

Schistosoma mansoni infection proceeds in normal mice in the absence of detectable levels of polyclonal or specific immunoglobulin (Ig)E until worms mature and deposit eggs. Hence, the course of a primary S. mansoni infection is not expected to vary appreciably in mice with defects in the IgE production. Experimental increase of IgE production early after infection may, however, influence worm development. In the first approach towards this goal, BALB/c mice were injected with interleukin(IL)4 to raise the level of endogenously synthesized IgE. A significant increase in serum polyclonal IgE and antischistosome IgG1 during the prepatent period was not associated with significant changes in worm and egg burden or liver pathology. During the second approach, mice were injected with IgE which was affinity purified from serum of BALB/c mice infected for 16 weeks with S. mansoni. The purified IgE bound to carbohydrate‐independent epitopes of soluble antigens from 3 h larvae, adult worms and eggs and recognized the schistosomular surface membrane. No differences in worm and egg load or granuloma number and size were noted between untreated and exogenous IgE‐injected mice. Together, the data demonstrate that by itself IgE does not influence the outcome of infection in primary murine S. mansoni.
Towards a ‘Human‐like’ Model of Tuberculosis: Intranasal Inoculation of LPS Induces Intragranulomatous Lung Necrosis in Mice Infected Aerogenically with Mycobacterium tuberculosis
Authors: Cardona P. J. , Llatjós R. , Gordillo S. , Díaz J. , Viñado B. , Ariza A. , Ausina V.
Published: March 2003

It is well known that one of the differences between murine and human tuberculosis is the lack of intragranulomatous necrosis in the former. The aim of this study was to create a feasible and reproducible model of an experimental model of murine tuberculosis in which this necrosis should be present. Considering the Shwartzman reaction as a possible explanation for intragranulomatous necrosis in human tuberculosis, C57Bl/6 mice, infected aerogenically with a virulent strain of Mycobacterium tuberculosis, were intranasally inoculated with lipopolysaccharide (LPS) on day 19 postinfection (p.i.). Twenty‐four hours later, neutrophils infiltrated the lung parenchyma in a significant level, and 10 days after necrosis could be detected in the centres of primary granulomas, that showed scanty macrophages and large amounts of collagen on an eosinophilic background. On the other hand, a significant decrease in the concentration of colony forming units (CFU) could be appreciated 24 h after the LPS inoculation. Afterwards, nonbronchogenic spreading of granulomas increased and higher levels of interferon (IFN)‐γ mRNA were detected. These results lend support to the Shwartzman reaction as the origin of the intragranulomatous necrosis in the M. tuberculosis infection, and provides a useful tool to improve experimental murine models in tuberculosis.
Inherited Complete Factor I Deficiency Associated with Systemic Lupus Erythematosus, Higher Susceptibility to Infection and Low Levels of Factor H
Authors: Amadei N. , Baracho G. V. , Nudelman V. , Bastos W. , Florido M. P. C. , Isaac L.
Published: March 2003

Here we describe two new cases of complete deficiency of factor I (fI) in two sisters from a consanguineous Brazilian family. The eldest sibling (20‐year‐old) developed systemic lupus erythematosus (SLE) early during childhood while the youngest had been committed on several occasions owing to repeated infections although she was asymptomatic for auto‐immune diseases. We also detected lower concentrations of C3 and factor B in both sisters. Biological functions dependent on complement activation such as the production of opsonins and killing of phagocytozed micro‐organisms, chemotactic factors and haemolytic activity were all significantly reduced in both probands. Consistent with the absence of fI and low levels of fH, a deregulated production of C3b was observed by bidimensional electrophoresis in sera of both the probands.
Blood γδ T Cells and γδ TCR V Gene Specificities in a Single Missense Mutation (L→Q271) in the Common Gamma Chain Gene
Authors: Schmalstieg F. C. , Palkowetz K. H. , Rudloff H. E. , Goldman A. S.
Published: March 2003

The numbers of blood CD4+, CD8+, and CD4−CD8− T cells bearing αβ T‐cell receptor (TCR) or γδ TCR molecules in males with a single missense mutation (L→Q271) in the common gamma chain gene (γc) were investigated by flow cytometry. Virtually all XCIDL→Q271 blood T cells that were CD4+ or CD8+ displayed αβ TCR but no γδ TCR. In contrast, CD4−CD8− T cells from affected males usually displayed γδ TCR, but no αβ TCR. The γδ TCR specificities were also studied. Except for the oldest subject, there was a direct relationship between blood CD3+ T cells that displayed γδ TCR and Vγ9 and Vδ2a specificities. Relative frequencies of CD3+ blood T cells that were Vγ9+ or Vδ2a+ were inversely related to age. In the oldest patient, the only detected γδ TCR specificity was Vδ1. Thus, in contrast to mice with no γc, XCIDL→Q271 blood T cells that bear γδ TCR with Vγ9/Vδ2a specificities develop but then decline in late childhood and thereafter. TCR with the Vδ1 specificity then appear in older survivors with XCIDL→Q271.
Modulation of IgD and CD20 by Ligation of CD5 on Tonsillar B Cells
Authors: Morikawa K. , Torii I. , Morikawa S.
Published: March 2003

The CD5 molecule, pan T cell marker, has been known to be expressed on a minor population of B cells, termed B‐1 cells. However, the physiological function and pathological role of CD5+B (B‐1) cells remain to be fully elucidated in humans. In the present study, we aimed to clarify the significance of CD5 expression on the B lymphocytes in human tonsil. Using flow cytometric analysis by three‐colour immunofluorescence staining, we observed a majority of the cell surface CD5‐positive (sCD5+) B cells among the sIgD+ B‐cell population, as previously described. Contrary to our expectation, approximately half of the sIgD+/sCD5+ B cells expressed CD38 on their cell surface. Furthermore, a small number of sCD5+ were observed in the sIgD− B cell population. The addition of anti‐CD5 monoclonal antibody (MoAb) to the culture induced downmodulation of sCD20 and sIgD of the tonsillar B cells, resulting in an increase of sCD38−/sIgD− (memory) B cells during the 10 day culture periods in the CD40/l cell culture system. Our findings suggest that ligation of CD5 might transduce the signal to regulate B cell maturation.
NK Cells, MHC Class I Molecules and the Missing Self
Authors: Kärre K.
Published: March 2003

This article is based on a lecture presented at the Novartis Prize ceremony at the International Congress of Immunology in July 2001. It gives a personal and historical perspective on the research performed by the author and his colleagues during the development and pursuit of the model of ‘missing‐self recognition’ for natural killer (NK) cells. This model is based on the idea that one important function of NK cells is to detect and eliminate cells because they fail to express normal self markers. Further mechanistic models predicted the existence of inhibitory major histocompatibility complex (MHC) class I specific receptors, later identified by the fellow Novartis laureates contributing in this issue. The article covers the first decade (1980–1990) of research on this concept. It discusses factors contributing to the formulation of a hypothesis, the use of predictions and experimental test models, the importance of international collaborations and reagent exchange, and several other aspects that allowed the progression of this research project. Finally, the perspective of today’s knowledge is used to discuss some surprising findings where the missing‐self hypothesis made the wrong predictions, or at least failed to make the correct ones.
Establishment of Animal Model of Antigen‐Specific T Lymphocyte Recruitment into Nasal Mucosa
Authors: KANAIZUMI E. , SHIRASAKI H. , SATO J. , WATANABE K. , HIMI T.
Published: March 2003

DO11.10 transgenic mice, expressing an ovalbumin (OVA)‐specific αβ T‐cell receptor (TCR), have been used as a model of various immune diseases associated with T lymphocytes. Some studies of immunoresponse in lung have involved adoptive transfer of DO11.10 mice. As of yet, however, there have been no studies of the adoptive transfer model in the upper airway. The purpose of this study was to establish an animal model to clarify the recruitment mechanism and the roles of Th2 cells in allergic rhinitis. In accordance with the adoptive transfer system, we generated Th0, Th1 and Th2 cells from DO11.10 mice and transferred them into wild type BALB/c mice. Following nasal OVA challenge to DO11.10 mice or to the BALB/c mice into which antigen‐specific Th2 cells had been transferred, the number of local antigen‐specific TCR‐positive cells accompanying the local eosinophilia had significantly increased. However, nasal OVA challenge to BALB/c mice into which antigen‐specific Th0 or Th1 cells were transferred failed to increase the number of local OVA‐specific TCR positive cells. These observations suggest that an antigen‐specific homing mechanism of Th2 cells may exist in nasal mucosa. Analysis of this model will assist in the development of new therapeutic strategy, which targets Th2 cells in allergic rhinitis.
Selective Small Antigenic Structures are Capable of Inducing Widespread Autoimmunity which Closely Mimics the Humoral Fine Specificity of Human SLE
Authors: MCCLAIN M. T. , SCOFIELD R. H. , KURIEN B. T. , GROSS T. F. , JAMES J. A.
Published: March 2003

Recent data have suggested that autoantibodies in lupus can progress from simple immunity against a few antigenic structures to a complex response against multiple autoantigens. Our aim was to determine whether these diverse epitope patterns can indeed be generated by antigenic challenge with a single, small structure. Rabbits were immunized with either a 60 kDa Ro peptide commonly antigenic in human systemic lupus erythematosus (SLE) (Ro 274–289) or one which is rarely a humoral target (Ro 500–515). Rabbits immunized with the antigenic peptide (Ro 274–289) not only developed antibodies to multiple epitopes of 60 kDa Ro and La, as has been described, but also produced non‐cross‐reactive antibodies to the common spliceosomal proteins Sm B′ and D1, and nRNP A and C. Rabbits immunized with the Ro 274–289 peptide also mount a progressive, diversified immune response to the sequential antigenic regions of these proteins (60 kDa Ro, Sm B′ and D1, nRNP A and C), which is nearly identical to that seen in human SLE. Animals immunized with the nonantigenic peptide Ro 500–515 develop antibodies only to 60 kDa Ro. These results demonstrate that loss of tolerance to select single, small antigenic structures can begin a cascade which virtually recreates, at the epitope level, the humoral autoimmune specificity seen in human SLE.
Anti‐Inflammatory Activities of Human Lactoferrin in Acute Dextran Sulphate‐Induced Colitis in Mice
Authors: Håversen L. A. , Baltzer L. , Dolphin G. , Hanson L. Å. , Mattsby‐Baltzer I.
Published: March 2003

In this study, we investigated the anti‐inflammatory effects of orally administered human lactoferrin (hLF) and two peptides, based on the bactericidal region of hLF (HLD1 and HLD2), on the course of experimental colitis. Acute colitis was induced in C57Bl/6 mice by giving 5% dextran sulphate (DX) in the drinking water. The mice were killed after 2 or 7 days of DX exposure. The animals were given hLF or the peptides orally twice a day (2 mg/dose/mouse) during the DX exposure. In the control animals, the hLF or the peptides were replaced by bovine serum albumin or water.

The appearance of occult blood in the faeces and macroscopic rectal bleeding were significantly delayed and partly reduced in the hLF‐treated animals compared with the control animals. The shortening of the colon, a pathological effect of DX exposure, was significantly less pronounced in the hLF‐treated group compared with the control group. Also, the interleukin‐1β (IL‐1β) levels in the blood were significantly diminished in this group after 2 days of DX exposure. A significantly lower crypt score was observed in the distal part of the colon in the hLF‐treated group compared with the control group. Also, significantly reduced numbers of CD4 cells, F4/80‐positive macrophages and tumour necrosis factor‐α‐producing cells were detected by immunohistochemistry in the distal colon of the hLF‐treated animals compared with the control animals after 7 days of DX exposure. A reduction was also observed concerning the IL‐10‐producing cells in the middle colonic submucosa. The HLD1 and HLD2 treatment, which was carried out for 2 days, only gave results almost identical to those of hLF, concerning clinical parameters after the 2 days of DX exposure. An even stronger effect was observed for HLD2, regarding decreased occult blood in the faeces and colon length.

Our results show that perorally given hLF mediates anti‐inflammatory effects on the DX‐induced acute colitis, and further suggest that the bactericidal region of the hLF molecule may be involved in these activities.
Effects of Various Adjuvants and a Viral Infection on the Antibody Specificity Toward Native or Cryptic Epitopes of a Protein Antigen
Authors: Gómez K. A. , Longhi S. A. , Marino V. J. , Mathieu P. A. , Loureiro M. E. , Coutelier J.‐P. , Roguin L. P. , Retegui L. A.
Published: March 2003

An immunization protocol that induces antibodies (Abs) directed to cryptic epitopes of a protein antigen (Ag) reduces the efficacy of vaccines that ideally should induce Abs against native epitopes. We have shown earlier that viral infections concomitant with immunization against a protein tend to shift the Ab specificity toward cryptic epitopes and tend to induce the production of autoantibodies (autoAbs). Here, we show the effects of three adjuvants on the Ab specificity in the absence or presence of a viral infection (lactate dehydrogenase‐elevating virus or LDV), with human growth hormone (hGH) being, as before, the protein Ag.

Pathogen‐free CBA/Ht and BALB/c mice were immunized with hGH in the presence of complete Freund's adjuvant (CFA), monophosphoryl lipid A (MPL) or alum, with the animals being either infected with LDV or not infected with LDV. Conventional and competition enzyme‐linked immunosorbent assays (ELISAs) indicated that in noninfected mice, CFA induced higher titres of anti‐hGH Ab than did MPL or alum, with the Ab being almost totally directed to cryptic hGH epitopes. Strikingly, CFA plus LDV infection in CBA/Ht mice shifted the specificity of the anti‐hGH Ab toward native epitopes, whereas the virus decreased the Ab titre when MPL or alum was used. Our Western blot results showed that 70% of mice immunized with hGH in the presence of any adjuvant produced autoAbs against a variety of tissue Ags. The amount of autoAb and the concentration of Ab to hGH cryptic epitopes did correlate, suggesting a relationship between both kinds of Ab. Significant differences were observed in the various effects of adjuvants and the viral infection between the two mouse strains used in this work.
Indoleamine 2,3‐Dioxygenase Expression in Patients with Acute Graft‐versus‐Host Disease after Allogeneic Stem Cell Transplantation and in Pregnant Women: Association with the Induction of Allogeneic Immune Tolerance?
Authors: Steckel N. K. , Kuhn U. , Beelen D. W. , Elmaagacli A. H.
Published: March 2003

Indoleamine 2,3‐dioxygenase (IDO) is an interferon‐γ (IFN‐γ)‐induced enzyme, which is suggested to play an important role in the prevention of allogeneic fetal rejection. IDO effects the suppression of T‐cell activity by catabolizing the essential amino acid l‐tryptophan. We studied IDO expression by reverse transcription polymerase chain reaction (RT‐PCR) in dendritic cells and by real‐time RT‐PCR in monocytes of patients undergoing allogeneic transplantation for leukaemia, who developed acute graft‐versus‐host disease (aGvHD), and compared the IDO expression with that of pregnant women and healthy volunteers.

A spontaneous IDO expression was detected in the monocytes of 20 pregnant women with an IDO/glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) ratio at a median of 1.0%, whereas none of 15 healthy volunteers or patients after allogeneic transplant had any detectable spontaneous IDO expression. The IDO expression increased by in vitro IFN‐γ stimulation in pregnant women (median 116%), healthy volunteers (median 11.7%) and patients with a low‐grade aGvHD (grades 0–II) 28 days after transplant (median 433%) but not in patients with a severe aGvHD (grades III–IV) (median 0%), which was highly significant (P < 0.01).

IDO expression was also measured in dendritic cells by qualitative RT‐PCR, where a spontaneous IDO expression was detected in 16 of 31 (52%) pregnant women versus none of 17 healthy volunteers and none of 62 studied patients after transplant. IFN‐γ‐induced IDO expression was detected in all pregnant women, all volunteers and 47 of 49 (96%) patients with a low‐grade aGvHD (grades 0–II) after transplant, whereas only in two of 13 (16%) patients with aGvHD grade III–IV was IFN‐γ‐induced IDO expression observed.

These data suggest that IDO expression might be involved in the development of allogeneic immune tolerance.
Sea level controls on the textural characteristics and depositional architecture of the Hueneme and associated submarine fan systems, Santa Monica Basin, California
Authors: Normark  , Piper  , Hiscott
Published: March 2003

Hueneme and Dume submarine fans in Santa Monica Basin consist of sandy channel and muddy levee facies on the upper fan, lenticular sand sheets on the middle fan, and thinly bedded turbidite and hemipelagic facies elsewhere. Fifteen widely correlatable key seismic reflections in high‐resolution airgun and deep‐towed boomer profiles subdivide the fan and basin deposits into time‐slices that show different thickness and seismic‐facies distributions, inferred to result from changes in Quaternary sea level and sediment supply. At times of low sea level, highly efficient turbidity currents generated by hyperpycnal flows or sediment failures at river deltas carry sand well out onto the middle‐fan area. Thick, muddy flows formed rapidly prograding high levees mainly on the western (right‐hand) side of three valleys that fed Hueneme fan at different times; the most recently active of the lowstand fan valleys, Hueneme fan valley, now heads in Hueneme Canyon. At times of high sea level, fans receive sand from submarine canyons that intercept littoral‐drift cells and mixed sediment from earthquake‐triggered slumps. Turbidity currents are confined to ‘underfit’ talweg channels in fan valleys and to steep, small, basin‐margin fans like Dume fan. Mud is effectively separated from sand at high sea level and moves basinward across the shelf in plumes and in storm‐generated lutite flows, contributing to a basin‐floor blanket that is locally thicker than contemporary fan deposits and that onlaps older fans at the basin margin. The infilling of Santa Monica Basin has involved both fan and basin‐floor aggradation accompanied by landward and basinward facies shifts. Progradation was restricted to the downslope growth of high muddy levees and the periodic basinward advance of the toe of the steeper and sandier Dume fan. Although the region is tectonically active, major sedimentation changes can be related to eustatic sea‐level changes. The primary controls on facies shifts and fan growth appear to be an interplay of texture of source sediment, the efficiency with which turbidity currents transport sand, and the effects of delta distributary switching, all of which reflect sea‐level changes.