ABSTRACT
Impatiens glandulifera, or Himalayan balsam, is a prolific invader of riverine habitats. Introduced from the Himalayas for ornamental purposes in 1839, this annual species has naturalised across Great Britain (GB) forming dense monocultures with negative affects across whole ecosystems. In 2006 a programme exploring biocontrol as an alternative control method was initiated and to date, two strains of the rust fungus Puccinia komarovii var. glanduliferae have been released. To better understand the observed differences in susceptibility of GB Himalayan balsam stands to the two rust strains, inoculation studies were conducted using urediniospores and basidiospores. Experiments revealed large variation in the susceptibility of stands to urediniospores of the two rust strains, with some resistant to both. Furthermore, the infectivity of basidiospores was found to differ, with some stands fully susceptible to the urediniospore stage, being immune to basidiospore infection. Therefore, before further rust releases at new sites, it is necessary to ensure complete compatibility of the invasive stands with both urediniospores and basidiospores. However, for successful control across GB it is essential that plant biotypes are matched to the most virulent rust strains. This will involve additional strains from the native range to tackle those biotypes resistant to the strains currently released.
Subject(s)
Biological Control Agents , Impatiens , Puccinia , Ecosystem , Impatiens/microbiology , Plant Diseases/microbiology , Plant Diseases/prevention & control , Puccinia/pathogenicity , Puccinia/physiology , United KingdomABSTRACT
Treatment with gold in the form of aurothiomaleate, silver or mercury (Hg) in genetically susceptible mouse strains (H-2(s)) induces a systemic autoimmune condition characterized by anti-nuclear antibodies targeting the 34-kDa nucleolar protein fibrillarin, as well as lymphoproliferation and systemic immune-complex (IC) deposits. In this study we have examined the effect of single-gene deletions for interferon (IFN)-gamma, interleukin (IL)-4, IL-6 or CD28 in B10.S (H-2(s)) mice on heavy metal-induced autoimmunity. Targeting of the genes for IFN-gamma, IL-6 or CD28 abrogated the development of both anti-fibrillarin antibodies (AFA) and IC deposits using a modest dose of Hg (130 microg Hg/kg body weight/day). Deletion of IL-4 severely reduced the IgG1 AFA induced by all three metals, left the total IgG AFA response intact, but abrogated the Hg-induced systemic IC deposits. In conclusion, intact IFN-gamma and CD28 genes are necessary for induction of AFA with all three metals and systemic IC deposits using Hg, while lack of IL-4 distinctly skews the metal-induced AFA response towards T helper type 1. In a previous study using a higher dose of Hg (415 microg Hg/kg body weight/day), IC deposits were preserved in IL-4(-/-) and IL-6(-/-) mice, and also AFA in the latter mice. Therefore, the attenuated autoimmunity following loss of IL-4 and IL-6 is dose-dependent, as higher doses of Hg are able to override the attenuation observed using lower doses.
Subject(s)
Autoimmune Diseases/chemically induced , CD28 Antigens/immunology , Cytokines/immunology , Metals, Heavy/toxicity , Animals , Antibodies, Antinuclear/immunology , Antigen-Antibody Complex , Autoimmune Diseases/immunology , CD28 Antigens/genetics , Chromosomal Proteins, Non-Histone/immunology , Dose-Response Relationship, Drug , Female , Gene Deletion , Genetic Predisposition to Disease , Gold/toxicity , Immunoglobulin G/immunology , Interferon-gamma/genetics , Interferon-gamma/immunology , Interleukin-4/immunology , Interleukin-6/genetics , Interleukin-6/immunology , Male , Mercury/toxicity , Mice , Mice, Knockout , Mice, Mutant Strains , Silver/toxicityABSTRACT
Natrium aurothiomaleate (GSTM) is a useful disease-modifying anti-rheumatic drug, but causes a variety of immune-mediated adverse effects in many patients. A murine model was used to study further the interaction of GSTM with the immune system, including induction of systemic autoimmunity. Mice were given weekly intramuscular injections of GSTM and controls equimolar amounts of sodium thiomaleate. The effects of gold on lymphocyte subpopulations were determined by flow cytometry. Humoral autoimmunity was measured by indirect immunofluorescence and immunoblotting, and deposition of immunoglobulin and C3 used to assess immunopathology. Gold, in the form of GSTM, stimulated the murine immune system causing strain-dependent lymphoproliferation and autoimmunity, including a major histocompatibility complex (MHC)-restricted autoantibody response against the nucleolar protein fibrillarin. GSTM did not cause glomerular or vessel wall IgG deposits. However, it did elicit a strong B cell-stimulating effect, including both T helper 1 (Th1)- and Th2-dependent isotypes. All these effects on the immune system were dependent on the MHC genotype, emphasizing the clinical observations of a strong genetic linkage for the major adverse immune reactions seen with GSTM treatment.
Subject(s)
Antirheumatic Agents/immunology , Autoimmunity , Gold Sodium Thiomalate/immunology , Animals , Antibodies, Antinuclear/biosynthesis , Antibodies, Antinuclear/blood , Autoimmunity/genetics , Female , Genetic Predisposition to Disease , Glomerular Mesangium/immunology , Immunoglobulin G/biosynthesis , Immunoglobulin G/blood , Immunoglobulin M/biosynthesis , Immunophenotyping , Lymphocyte Subsets/immunology , Mice , Mice, Inbred Strains , Species Specificity , Spleen/immunologyABSTRACT
F1 hybrids of SWR (H-2(q)) and SJL (H-2(s)) mice spontaneously develop a lupuslike condition in an age-dependent manner, and these two H-2 haplotypes also confer susceptibility to induction of systemic autoimmunity by heavy metals such as mercury, silver, and gold with anti-fibrillarin antibodies (AFA) as marker. The aim of this study was to determine how the mixing of two susceptible genomes might influence expression of idiopathic and induced autoimmunity over a period of 14 mo of exposure to mercury and silver. Spontaneous autoimmunity first appeared as antinuclear antibodies (ANA) in females at 10 wk of age and in males at 10 mo of age, and was followed by development of anti-chromatin antibodies. Antibodies to double-stranded DNA developed in 60% of males and 20% of females. Thirty percent of males and 10% of females developed a coarsely speckled ANA pattern associated with high titers of anti-Sm antibodies. Glomerular immune complex (IC) deposits and a proliferative glomerulonephritis were seen at 17 mo of age. The F1 hybrids treated with metals showed no exaggeration of spontaneous autoimmunity. However, the metals suppressed the spontaneous development of anti-Sm and antichromatin antibodies. The metal-induced AFA, linked to the H-2(s) and H-2(q) haplotype, reached a maximum after 3-4 mo of treatment and then declined; 33% of the silver-treated hybrids finally became AFA-negative, despite continuous treatment. The decline in ANoA during metal treatment is contrary to the situation in metal-treated SJL mice. This indicates that dominant SWR background genes suppressed induction of certain autoimmune traits in the (SWR x SJL)F1 hybrid mice.
Subject(s)
Antibodies, Antinuclear/analysis , Autoimmunity , Mercury/toxicity , Silver/toxicity , Age Factors , Animals , Antibody Formation , Female , Genetic Predisposition to Disease , Genome , Glomerulonephritis/chemically induced , Male , Mice , Mice, Inbred StrainsABSTRACT
The possible health effects of the organic mercury compound thimerosal (ethylmercurithiosalicylate), which is rapidly metabolized to ethylmercury (EtHg), have recently been much debated and the effect of this compound on the immune system is largely unknown. We therefore studied the effect of thimerosal by treating A.SW (H-2s) mice, susceptible to induction of autoimmunity by heavy metals, with 10 mg thimerosal/L drinking water (internal dose ca 590 microg Hg/kg body weight/day) for up to 30 days. The lymph node expression of IL-2 and IL-15 mRNA was increased after 2 days, and of IL-4 and IFN-gamma mRNA after 6 and 14 days. During the first 14 days treatment, the number of splenocytes, including T and B cells as well as Ig-secreting cells decreased. A strong immunostimulation superseded after 30 days treatment with increase in splenic weight, number of splenocytes including T and B cells and Ig-secreting cells, and Th2- as well as Th-1-dependent serum immunoglobulins. Antinucleolar antibodies (ANoA) targeting the 34-kDa nucleolar protein fibrillarin, and systemic immune-complex deposits developed. The H-2s strains SJL and B10.S also responded to thimerosal treatment with ANoA. The A.TL and B10.TL strain, sharing background genes with the A.SW and B10.S strain, respectively, but with a different H-2 haplotype (t1), did not develop ANoA, linking the susceptibility to H-2. Thimerosal-treated H-2s mice homozygous for the nu mutation (SJL-nu/nu), or lacking the T-cell co-stimulatory molecule CD28 (B10.S-CD28-/-), did not develop ANoA, which showed that the autoimmune response is T-cell dependent. Using H-2s strains with targeted mutations, we found that IFN-gamma and IL-6, but not IL-4, is important for induction of ANoA by thimerosal. The maximum added renal concentration of thimerosal (EtHg) and inorganic mercury occurred after 14 days treatment and was 81 microg Hg/g. EtHg made up 59% and inorganic mercury 41% of the renal mercury. In conclusion, the organic mercury compound thimerosal (EtHg) has initial immunosuppressive effects similar to those of MeHg. However, in contrast to MeHg, thimerosal treatment leads in genetically susceptible mice to a second phase with strong immunostimulation and autoimmunity, which is T-cell dependent, H-2 linked and may at least partly be due to the inorganic mercury derived from the metabolism of ethyl mercury.
Subject(s)
Autoimmunity , Immunosuppressive Agents/immunology , Thimerosal/immunology , Thimerosal/pharmacology , Administration, Oral , Animals , Antibodies, Antinuclear/blood , Antibodies, Antinuclear/drug effects , Antibodies, Antinuclear/immunology , B-Lymphocytes/drug effects , B-Lymphocytes/immunology , B-Lymphocytes/physiology , B7-1 Antigen/metabolism , Blood Vessels/chemistry , Blood Vessels/drug effects , CD4-Positive T-Lymphocytes/drug effects , CD4-Positive T-Lymphocytes/immunology , Cell Communication/drug effects , Cell Communication/immunology , Cell Proliferation/drug effects , Female , Gene Expression/drug effects , Gene Expression/genetics , Immunoglobulin G/chemistry , Immunoglobulin G/drug effects , Immunoglobulin Light Chains/blood , Immunoglobulin Light Chains/drug effects , Immunoglobulin Light Chains/immunology , Immunosuppressive Agents/pharmacology , Interferon-gamma/genetics , Interferon-gamma/immunology , Interferon-gamma/metabolism , Interleukin-4/genetics , Interleukin-4/immunology , Interleukin-4/metabolism , Kidney/blood supply , Kidney/chemistry , Kidney/drug effects , Leukocytes, Mononuclear/drug effects , Leukocytes, Mononuclear/immunology , Lymph Nodes/chemistry , Lymph Nodes/drug effects , Lymph Nodes/metabolism , Lymphocyte Activation/drug effects , Lymphocyte Activation/immunology , Male , Mesentery/drug effects , Mesentery/metabolism , Mice , Mice, Transgenic , Mutagenesis, Site-Directed/genetics , Mutagenesis, Site-Directed/immunology , Mutation/drug effects , Organ Size/drug effectsABSTRACT
The characterization of autoantibody specificities in rheumatic diseases is important in both diagnostic and basic research areas. Identification of the epitopes recognized by autoantibodies and their clinical and biological significance is not a trivial task. Epitopes may range in complexity from simple linear sequences of amino acids to complex quaternary structures. In addition to this structural complexity the frequency with which an autoantigen and its epitopes are recognized in a patient population may be useful in diagnosis, defining disease subgroups, and may offer information on disease prognosis. In this review recent advances in the epitope mapping of autoantigens in connective tissue diseases are discussed, with particular emphasis placed on the methodologies used to identify epitopes and the classification of the structural features of epitopes. To illustrate the identification of epitope structure, clinically relevant autoantigens, including CENP-A, PM/Scl-100, fibrillarin, filaggrin, Ro-52, and dsDNA, are discussed as examples of each type of epitope.
Subject(s)
Autoantigens/immunology , Connective Tissue Diseases/immunology , Epitope Mapping/methods , Epitopes, B-Lymphocyte/analysis , Amino Acid Sequence , Animals , Epitopes, B-Lymphocyte/immunology , Filaggrin Proteins , Humans , Molecular Mimicry/immunology , Molecular Sequence DataABSTRACT
Exposure of SJL/J mice to mercury induces an anti-nucleolar autoantibody response. The predominant target is fibrillarin, a 34-kDa component of the small nucleolar ribonucleoprotein particles (snoRNP), but other proteins are also recognized. To characterize these proteins, monoclonal IgG anti-nucleolar antibodies were produced from HgC12-treated SJL/J mice. One monoclonal, 17C12, recognized fibrillarin, while two others, 7G3 and 6G10, were found to immunoprecipitate snoRNP particles but not fibrillarin. Antibody 6G10 gave a nucleolar immunofluorescence pattern in human, murine, and amphibian cells, but was negative in immunoblot. The 7G3 monoclone reacted with a 60-kDa protein conserved in human and murine, but not amphibian, cell lines. The 7G3 and 6G10 antigens and fibrillarin colocalized to the nucleolus and Cajal bodies in interphase cells and decorated metaphase chromosomes. These studies suggest that the mercury-induced anti-nucleolar antibody response targets other protein components of the snoRNP particles in addition to fibrillarin.
Subject(s)
Antibodies, Antinuclear/immunology , Autoimmunity , Chromosomal Proteins, Non-Histone/immunology , Ribonucleoproteins, Small Nucleolar/immunology , Xenobiotics/pharmacology , 3T3 Cells , Animals , Antibodies, Monoclonal/immunology , Cell Cycle , Cell Line , Cell Nucleolus/metabolism , Coiled Bodies/chemistry , Female , HeLa Cells , Humans , Mercuric Chloride/pharmacology , Mice , Microscopy, Fluorescence , Rats , Ribonucleoproteins, Small Nucleolar/metabolismABSTRACT
Although evidence indicates that environmental factors play a major role in precipitating systemic autoimmunity in genetically susceptible individuals, little is known about the mechanisms involved. Certain heavy metals, such as mercury, are potent environmental immunostimulants that produce a number of immunopathologic sequelae, including lymphoproliferation, hypergammaglobulinemia, and overt systemic autoimmunity. Predisposition to such metal-induced immunopathology has been shown to be influenced by both MHC and non-MHC genes, as well as susceptibility to spontaneous lupus, in mice and other experimental animals. Among the various mouse strains examined to date, the DBA/2 appears to uniquely lack susceptibility to mercury-induced autoimmunity (HgIA), despite expressing a susceptible H-2 haplotype (H-2d). To define the genetic basis for this trait, two genome-wide scans were conducted using F2 intercrosses of the DBA/2 strain with either the SJL or NZB strains, both of which are highly susceptible to HgIA. A single major quantitative trait locus on chromosome 1, designated Hmr1, was shown to be common to both crosses and encompassed a region containing several lupus susceptibility loci. Hmr1 was linked to glomerular immune complex deposits and not autoantibody production, suggesting that DBA/2 resistance to HgIA may primarily involve the later stages of disease pathogenesis. Identification and characterization of susceptibility/resistance genes and mechanisms relevant to the immunopathogenesis of mercury-induced autoimmunity should provide important insights into the pathogenesis of autoimmunity and may reveal novel targets for intervention.
Subject(s)
Autoimmune Diseases/genetics , Chromosome Mapping , Immunity, Innate/genetics , Mercuric Chloride/immunology , Xenobiotics/immunology , Animals , Autoimmune Diseases/chemically induced , Autoimmune Diseases/immunology , Chromosome Mapping/methods , Crosses, Genetic , Female , Genetic Linkage/immunology , Genetic Markers/immunology , Genetic Predisposition to Disease/genetics , Mice , Mice, Inbred DBA , Mice, Inbred NZB , Mice, Inbred Strains , Quantitative Trait, Heritable , Species SpecificityABSTRACT
The aims of this study were to compare the in vitro responses of murine lymphocytes to HgCl2 to determine the requirement for adherent cells, and the contribution that costimulation plays in T cell proliferation. The in vitro proliferative response of murine splenocytes to HgCl2 was found to be both cell concentration- and HgCl2 concentration-dependent with the greatest response occurring with 5 x 10(6) cells/ml in the presence of 10(-5) M HgCl2. Both CD4+ and CD8+ T cells proliferated in response to HgCl2, but B cells and immature T cells (thymocytes) did not. Proliferation required the presence of splenic adherent cells and was inhibited by addition of anti-IL-1 alpha antibodies. Antibodies to the other co-stimulatory molecules CD40 ligand, CD80 (B7-1), and CD86 (B7-2), although inhibitory, were less effective. Xenobiotics such as the heavy metal mercury can elicit a spectrum of immunological responses ranging from immunosuppression to autoimmunity. The most common response, in vivo and in vitro, is lymphoproliferation, which may be a prelude to immune activation. Although a number of the requirements for mercury-induced T cell proliferation in vitro have been described, the role that adherent cells play remains to be explained. The studies described here show that interaction between co-stimulatory molecules of adherent cells and mature T cells contributes to HgCl2-induced T cell proliferation. Among these co-stimulatory molecules, IL-1 appears to play an important role. The requirement for mature T cells, adherent cells, and co-stimulatory molecules argues that HgCl2-induced T cell proliferation possesses the properties of an antigen-induced response.
Subject(s)
Interleukin-1/metabolism , Mercuric Chloride/toxicity , T-Lymphocytes/drug effects , T-Lymphocytes/immunology , Animals , Antibodies/pharmacology , Cell Differentiation , Cell Division/drug effects , Female , In Vitro Techniques , Interleukin-1/antagonists & inhibitors , Lymphocyte Activation/drug effects , Macrophages/drug effects , Macrophages/immunology , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Inbred DBA , T-Lymphocytes/cytologyABSTRACT
The diverse genetic backgrounds of lupus-prone murine models, which produce both quantitative and qualitative differences in disease expression, may be a valuable resource for studying the influence of environmental exposure on autoimmune disease in sensitive populations. We tested this premise by exposing autoimmune-prone BXSB and the nonautoimmune C57BL/6 mice to the heavy metal mercury. Although both strains express a nonsusceptible H-2 haplotype, exposure to mercury accelerated systemic autoimmunity in both male and female BXSB mice, whereas the C57BL/6 mice were resistant. The subclasses of antichromatin antibodies elicited in BXSB mice by mercury exposure were more consistent with the predominant Th1-type response of idiopathic disease than with the Th2-type response found in mercury-induced autoimmunity (HgIA). The appearance and magnitude of both humoral and cellular features of systemic autoimmunity correlated with the mercury dose. Furthermore, environmentally relevant tissue levels of mercury were associated with exacerbated systemic autoimmunity. These studies demonstrate that xenobiotic exposure can accelerate spontaneous systemic autoimmunity, and they support the possibility that low-level xenobiotic exposure enhances susceptibility to systemic autoimmunity in genetically susceptible individuals.
Subject(s)
Genetic Predisposition to Disease , Lupus Erythematosus, Systemic/chemically induced , Mercury/adverse effects , Xenobiotics/adverse effects , Animals , Antibody Formation , Autoantibodies/immunology , Autoimmunity , Chromatin/immunology , Disease Models, Animal , Female , Lupus Erythematosus, Systemic/genetics , Lupus Erythematosus, Systemic/immunology , Male , Mice , Mice, Inbred C57BLABSTRACT
The heavy metal mercury elicits a genetically restricted autoantibody response in mice that targets the nucleolar autoantigen fibrillarin. HgCl2-induced cell death of macrophages resulted in the proteolytic cleavage of fibrillarin. A prominent feature of mercury-induced cell death was the generation of a 19-kDa fragment of fibrillarin that was not found following apoptotic or nonapoptotic cell death induced by stimuli other than mercury. Proteolysis of fibrillarin lacking cysteines, and therefore unable to bind mercury, also produced the 19-kDa fragment, suggesting that a mercury-fibrillarin interaction was not necessary for the unique cleavage pattern of this self-Ag. In contrast to immunization with full-length fibrillarin, the 19-kDa fragment produced anti-fibrillarin Abs with some of the properties of the HgCl2-induced anti-fibrillarin response. We propose that cell death following exposure to an autoimmunity-inducing xenobiotic can lead to the generation of novel protein fragments that may serve as sources of antigenic determinants for self-reactive T lymphocytes.
Subject(s)
Apoptosis/drug effects , Apoptosis/immunology , Autoantigens/metabolism , Peptide Fragments/biosynthesis , Peptide Fragments/immunology , Xenobiotics/pharmacology , Animals , Apoptosis/genetics , Autoantibodies/biosynthesis , Autoantigens/administration & dosage , Autoantigens/genetics , Autoantigens/immunology , Cell Line , Chromosomal Proteins, Non-Histone/administration & dosage , Chromosomal Proteins, Non-Histone/genetics , Chromosomal Proteins, Non-Histone/immunology , Chromosomal Proteins, Non-Histone/metabolism , Hydrolysis , Injections, Subcutaneous , Macrophages/cytology , Macrophages/drug effects , Macrophages/immunology , Macrophages/metabolism , Mercuric Chloride/pharmacology , Mice , Mice, Inbred C57BL , Molecular Weight , Mutagenesis, Site-Directed , Peptide Fragments/administration & dosage , Peptide Fragments/geneticsABSTRACT
Arginine methylation in RNA-binding proteins containing arginine- and glycine-rich RGG motifs is catalyzed by specific protein arginine N-methyltransferase in cells. We previously showed that lymphoblastoid cells grown in the presence of an indirect methyltransferase inhibitor, adenosine dialdehyde (AdOx), accumulated high level of hypomethylated protein substrates for the endogenous protein methyltransferases or recombinant yeast arginine methyltransferase [Li, C. et al. (1998) Arch. Biochem. Biophys. 351, 53-59]. In this study we fractionated the lymphoblastoid cells to locate the methyltransferases and the substrates in cells. Different sets of hypomethylated methyl-accepting polypeptides with wide range of molecular masses were present in cytosolic, ribosomal, and nucleus fractions. The methylated amino acid residues of the methyl-accepting proteins in these fractions were determined. In all three fractions, dimethylarginine was the most abundant methylated amino acid. The protein-arginine methyltransferase activities in the three fractions were analyzed using recombinant fibrillarin (a nucleolar RGG protein) as the methyl-accepting substrate. Fibrillarin methylation was strongest in the presence of the cytosolic fraction, followed by the ribosomal and then the nucleus fractions. The results demonstrated that protein-arginine methyltransferases as well as their methyl-accepting substrates were widely distributed in different subcellular fractions of lymphoblastoid cells.
Subject(s)
Arginine/metabolism , Lymphocytes/metabolism , Peptides/metabolism , Protein-Arginine N-Methyltransferases/metabolism , Adenosine/analogs & derivatives , Adenosine/pharmacology , Blotting, Western , Enzyme Inhibitors/pharmacology , Glutathione Transferase/biosynthesis , Glutathione Transferase/genetics , Humans , Lymphocytes/drug effects , Methylation , Peptides/chemistry , Protein-Arginine N-Methyltransferases/antagonists & inhibitors , Protein-Arginine N-Methyltransferases/genetics , Recombinant Fusion Proteins , Subcellular FractionsABSTRACT
OBJECTIVE: Proteins that are phosphorylated during apoptosis are commonly precipitated by autoantibodies found in the sera of patients with systemic lupus erythematosus. We sought to determine whether scleroderma autoantigens such as small nucleolar RNPs (snoRNP) also associate with phosphoproteins in response to various cellular stressors. METHODS: We screened a panel of monoclonal antibodies derived from mice exposed to mercury, a well-characterized murine model of the anti-snoRNP autoimmune response, for the ability to selectively precipitate phosphoproteins from radiolabeled lysates prepared from Jurkat T cells subjected to stressful stimuli. RESULTS: Monoclonal antibodies reactive with snoRNPs precipitated a phosphoprotein complex (pp42, pp34, and pp23) from lysates prepared from apoptotic cells. Several novel phosphoproteins (pp62 and pp18) were also observed. The phosphorylation and/or recruitment of these proteins to the snoRNP complex is induced by multiple apoptotic stimuli (e.g., Fas ligation, anisomycin, or ultraviolet irradiation), an effect that is blocked by overexpression of Bcl-2. We were unable to demonstrate an association of the phosphoprotein complex with snoRNPs in cells treated with the xenobiotic agent mercury. The snoRNP-associated phosphoprotein complex is composed of serine/arginine (SR) splicing factors, including SRp40. CONCLUSION: The association of phosphorylated SR proteins with snoRNPs in cells undergoing apoptosis suggests that the immune response to fibrillarin that characterizes a subset of patients with scleroderma may be related to cell death induced by apoptotic stimuli (e.g., Fas ligation, irradiation, or chemical toxins), or by exposure to mercury.
Subject(s)
Apoptosis/physiology , Autoantigens/immunology , Nuclear Proteins/immunology , Phosphoproteins/immunology , Ribonucleoproteins, Small Nuclear/immunology , Scleroderma, Systemic/immunology , Animals , Chromosomal Proteins, Non-Histone/metabolism , Humans , Jurkat Cells/drug effects , Mercury/pharmacology , Mice , Nuclear Proteins/chemistry , Nuclear Proteins/metabolism , Peptide Mapping , Phosphoproteins/chemistry , Phosphoproteins/metabolism , Phosphorylation/radiation effects , RNA Splicing , RNA-Binding Proteins , Ribonucleoproteins, Small Nuclear/metabolism , Serine-Arginine Splicing Factors , Ultraviolet RaysABSTRACT
The linkage between xenobiotic exposures and autoimmune diseases remains to be clearly defined. However, recent studies have raised the possibility that both genetic and environmental factors act synergistically at several stages or checkpoints to influence disease pathogenesis in susceptible populations. These observations predict that individuals susceptible to spontaneous autoimmunity should be more susceptible following xenobiotic exposure by virtue of the presence of predisposing background genes. To test this possibility, mouse strains with differing genetic susceptibility to murine lupus were examined for acceleration of autoimmune features characteristic of spontaneous systemic autoimmune disease following exposure to the immunostimulatory metals nickel and mercury. Although NiCl(2) exposure did not exacerbate autoimmunity, HgCl(2) significantly accelerated systemic disease in a strain-dependent manner. Mercury-exposed (NZB X NZW)F1 mice had accelerated lymphoid hyperplasia, hypergammaglobulinemia, autoantibodies, and immune complex deposits. Mercury also exacerbated immunopathologic manifestations in MRL+/+ and MR -lpr mice. However, there was less disease acceleration in lpr mice compared with MRL+/+ mice, likely due to the fact that environmental factors are less critical for disease induction when there is strong genetic susceptibility. Non-major histocompatibility complex genes also contributed to mercury-exacerbated disease, as the nonautoimmune AKR mice, which are H-2 identical with the MRL, showed less immunopathology than either the MRL/lpr or MRL+/+ strains. This study demonstrates that genetic susceptibility to spontaneous systemic autoimmunity can be a predisposing factor for HgCl(2)-induced exacerbation of autoimmunity. Such genetic predisposition may have to be considered when assessing the immunotoxicity of xenobiotics. Additional comparative studies using autoimmune-prone and nonautoimmune mice strains with different genetic backgrounds will help determine the contribution that xenobiotic exposure makes in rendering sensitive populations susceptible to autoimmune diseases.
Subject(s)
Autoimmunity/drug effects , Lupus Erythematosus, Systemic/etiology , Xenobiotics/toxicity , Animals , Autoantibodies/biosynthesis , Autoimmunity/genetics , Disease Models, Animal , Environmental Exposure , Female , Humans , Lupus Erythematosus, Systemic/genetics , Lupus Erythematosus, Systemic/immunology , Mercuric Chloride/toxicity , Mice , Mice, Inbred AKR , Mice, Inbred MRL lpr , Mice, Inbred NZB , Nickel/toxicity , Species SpecificityABSTRACT
Fibrillarin is a 34-kDa nucleolar protein associated with many of the small nucleolar ribonucleoprotein (snoRNP) particles and plays a role in ribosomal RNA processing. A subset of patients with the systemic autoimmune disease Scleroderma produce autoantibodies against fibrillarin and it is a genetically restricted target of murine mercury-induced autoimmunity. To aid in characterizing the antigenicity of fibrillarin, we have constructed two forms of mouse fibrillarin. The wild-type clone contains two cysteine residues that enable the protein to form an intramolecular disulfide bond, whereas the mutant clone contains alanine replacements which cannot form the disulfide bond. We have successfully expressed and purified both wild-type and mutant recombinant mouse fibrillarin using nickel-chelation chromatography. The combination of T7 promoter-driven expression vector pET28 and Escherichia coli strain JM109(DE3) induced at 25 degrees C yielded up to 19 mg of 94% pure recombinant protein per liter of culture. As the antigenicity of fibrillarin requires the full-length protein, the purification protocol was optimized for isolation of the full-length protein by the addition of N- and C-terminal T7 Tag and FLAG epitope sequences to the fibrillarin sequence. Anti-peptide antibodies were used in immunoblot to identify conditions favoring minimal proteolysis of recombinant protein. Both wild-type and mutant recombinant fibrillarin, purified under denaturing conditions and in the presence of 2-mercaptoethanol, were recognized by anti-fibrillarin antibodies from Scleroderma patients and exhibited structural similarities to eukaryotic and in vitro translated fibrillarin.
Subject(s)
Chromosomal Proteins, Non-Histone/genetics , Chromosomal Proteins, Non-Histone/isolation & purification , Amino Acid Sequence , Animals , Autoantigens/biosynthesis , Autoantigens/genetics , Autoantigens/isolation & purification , Base Sequence , Chromosomal Proteins, Non-Histone/biosynthesis , DNA, Recombinant/genetics , Disulfides/chemistry , Escherichia coli/genetics , Genetic Vectors , Humans , Mercuric Chloride , Mice , Mutagenesis, Site-Directed , Protein Conformation , Recombinant Proteins/biosynthesis , Recombinant Proteins/genetics , Recombinant Proteins/isolation & purificationABSTRACT
Imbalances of Th1- and Th2-type responses have been postulated to be a predisposing factor for both humoral and cellular mediated autoimmune diseases. To further define their roles in systemic autoimmunity, IL-4 and IFN-gamma gene knockout mice were studied for susceptibility to the prototypic Th2-mediated mercury-induced autoimmunity. A predominant Th2-type response following HgCl2 treatment of wild-type B10.S mice was confirmed by the findings of a significant increase in splenic IL-4 and hypergammaglobulinemia primarily of the IgG1 isotype, without an increase in IFN-gamma levels. Paradoxically, IL-4-deficient mice developed the characteristic anti-nucleolar autoantibodies and tissue deposition of immune complexes, while IFN-gamma-deficient mice had very low autoantibody levels and essentially normal immunohistology. Studies to define defects in Ab responses of IFN-gamma-deficient mice, using the T-dependent Ag (4-hydroxy-3-nitrophenyl)acetyl, revealed an attenuated IgG response to low and to a lesser extent high doses of (4-hydroxy-3-nitrophenyl)acetyl-hemocyanin, but maintenance of affinity maturation. These results indicate that Th1/Th2 imbalance does not directly play a role in susceptibility to mercury-induced autoimmunity, and suggest that the dependence on Th1-type responses in certain autoimmune diseases is due to the requirement for IFN-gamma for Ab production to weakly antigenic self molecules.
Subject(s)
Autoimmune Diseases/chemically induced , Interferon-gamma/physiology , Mercuric Chloride/immunology , Th1 Cells/immunology , Th2 Cells/immunology , Animals , Autoantibodies/biosynthesis , Autoimmune Diseases/genetics , Autoimmune Diseases/immunology , Disease Susceptibility , Dose-Response Relationship, Immunologic , Haptens/immunology , Interferon-gamma/genetics , Interleukin-4/biosynthesis , Interleukin-4/deficiency , Interleukin-4/genetics , Mice , Mice, Inbred C57BL , Mice, Knockout , Nitrophenols/immunology , Phenylacetates , Th1 Cells/metabolism , Th2 Cells/metabolismABSTRACT
Asians and Pacific Islanders (API) have an increasingly visible presence in the United States. This diverse population--encompassing persons with ancestry from East and Southeast Asia, the Indian subcontinent, and the Pacific islands--has grown at a faster rate than any other major racial or ethnic group. In 1996 Asian Americans numbered approximately 9.7 million (up from 3.8 million in 1980), nearly 4 percent of the U.S. population. The Census Bureau projects that this population group will reach 34.4 million by 2050, representing roughly 9 percent of all Americans. While immigration has fueled much of this growth, Asians' young age structure also will help boost their numbers in the next century. Fifty-six percent of Asian Americans live in three states--California, New York and Hawaii. Asian Americans comprise very small proportions of the populations of most other states. About 77 percent of the 2.8 million API households in 1996 were families, compared with 69 percent of white households. Roughly one in six Asian American households has five or more persons, compared with one in 12 white households. Educationally, Asians tend to be high achievers--42 percent of all API adults have at least a bachelor's degree, compared with 26 percent of while adults. Two-thirds of Asian Americans participated in the civilian labor force in 1996. Among employed Asians, one-third held managerial and professional jobs. Both proportions were roughly the same as for whites. Although the median income for API households was 9 percent higher than for white households in 1995, this difference is largely due to Asian households having more workers contributing to the household income. Despite these apparent measures of success, the poverty rates for Asian American families and individuals are nearly twice as high as those for whites.
Subject(s)
Asian/statistics & numerical data , Adult , Age Distribution , Aged , Censuses , Demography , Educational Status , Emigration and Immigration/statistics & numerical data , Employment/statistics & numerical data , Ethnicity/statistics & numerical data , Female , Humans , Income/statistics & numerical data , Male , Middle Aged , Occupations/statistics & numerical data , Pacific Islands/ethnology , Population Dynamics , Poverty/statistics & numerical data , Socioeconomic Factors , United StatesABSTRACT
The heavy metal mercury elicits a genetically restricted, anti-nucleolar autoantibody response that targets fibrillarin, a 34-kDa protein component of many small nucleolar ribonucleoprotein particles. The mechanisms by which a toxin such as mercury elicits an autoantibody response that predominantly targets a single intracellular protein autoantigen remain uncertain, but may be prefaced by mercury gaining access to the intracellular environment. Mercury-induced cell death was associated with loss of fibrillarin antigenicity and modification of the molecular properties of fibrillarin as revealed by aberrant migration under nonreducing conditions in SDS-PAGE. Addition of mercury to isolated nuclei also resulted in aberrant migration of fibrillarin, but not other nuclear autoantigens. The sensitivity of the HgCl2-induced modification of fibrillarin to 2-ME, iodoacetamide, and hydrogen peroxide suggested interaction of mercury with the two cysteines in the fibrillarin sequence. This was confirmed by mutation of the cysteines to alanines, which abolished the aberrant migration of fibrillarin in the presence of HgCl2. The modification of the molecular structure of fibrillarin by mercury reduced immunoprecipitation by anti-fibrillarin autoantibodies, pointing to unmodified fibrillarin as the B cell Ag and implicating mercury-modified fibrillarin as the source of T cell antigenicity. These observations demonstrate for the first time that an environmental toxin can alter the physicochemical properties of an autoantigen and may help to explain the antigenic specificity of mercury-induced murine autoimmunity.
Subject(s)
Autoantigens/drug effects , Autoantigens/immunology , Chromosomal Proteins, Non-Histone/immunology , Chromosomal Proteins, Non-Histone/pharmacology , Mercuric Chloride/immunology , Mercuric Chloride/pharmacology , Xenobiotics/immunology , Xenobiotics/pharmacology , Antibodies, Monoclonal/chemistry , Autoantibodies/metabolism , Binding Sites, Antibody , Cell Death/drug effects , Cell Death/immunology , Cell Nucleus/drug effects , Cell Nucleus/immunology , Chromosomal Proteins, Non-Histone/drug effects , Cysteine/physiology , Disulfides/chemistry , Electrophoresis, Polyacrylamide Gel , Epitopes/chemistry , Humans , Subcellular Fractions/drug effects , Subcellular Fractions/immunologySubject(s)
Autoimmunity/drug effects , Immune System/drug effects , Lymphoid Tissue/immunology , Mercury/toxicity , Animals , Drug Hypersensitivity/complications , Drug Hypersensitivity/epidemiology , Drug Synergism , Humans , Immunosuppression Therapy , Lymphocytes/drug effects , Lymphoid Tissue/drug effects , Metals/pharmacologyABSTRACT
Demographic changes have shaped the nation's past and will continue to shape its future. During the first half of the 1990s, the U.S. population grew, on average, by 2.7 million people each year, reaching 262.8 million in 1995. Population growth is projected to continue for the next 50 years, although at a slower rate. The forecast is for more than 390 million Americans by the year 2050. As the U.S. population grows, it will increasingly become more diverse along many socioeconomic dimensions. This increasing diversity will represent an historic shift in America's racial and ethnic composition with long-range implications for how we view racial issues, how we define racial categories and how the political landscape will be refashioned. By the middle of the 21st century the "minority" population will almost equal the size of the non-Hispanic white population. The minority population grew 14 percent during the first half of the 1990s compared with a 3 percent growth in the non-Hispanic white population. But even within the minority population, growth rates varied. Between 1990 and 1995, the Asian population grew 23 percent, the Hispanic population 20 percent and the African American population increased their numbers by 8 percent. Hispanics are projected to outnumber African Americans within the next 15 years. In part, these demographic changes are shifting because of U.S. immigration policies. Until the early 1960s, immigrants to the United States were primarily of white, European stock. Nowadays, Europeans account for about 20 percent of the immigrants. Three-quarters of legal immigrants in the mid-1990s now come from Latin America, the Caribbean and Asia. The increasing racial and ethnic diversity in the United States will create both challenges and opportunities for U.S. schools and businesses in the future. The magnitude of these numbers and their geographic location will be important factors to consider as we prepare for the 21st century.