One-off streptococcal serologic testing in young children with recurrent tonsillitis.

OBJECTIVES: Recurrent acute tonsillitis in children under 4 years of age is usually viral, making antibiotic therapy inappropriate and the indication for tonsillectomy uncertain. Identifying those young children with bacterial infections is therefore important. The purpose of this study was to determine whether one-off streptococcal serologic testing is a useful tool in assessing recurrent acute tonsillitis in young children. METHODS: We performed a retrospective study of 45 children (35 male and 10 female) under the age of 4 years who were found by a staff otolaryngologist to have recurrent acute tonsillitis over a 5-year period and had one-off serologic testing for anti-streptolysin O titers and anti-deoxyribonuclease B levels. Data were collected by chart review. RESULTS: Three children (6.7%) had clearly positive titers for either one or both streptococcal antibodies. Children with negative serologic results were significantly less likely to have shown a significant response to antibiotic therapy for their acute episodes (26% versus 100%; p = .026). Nine children (20%) eventually underwent tonsillectomy, all of whom had negative serologic results. CONCLUSIONS: Anti-streptolysin O and anti-deoxyribonuclease B levels may aid clinical evaluation of recurrent acute tonsillitis in young children in differentiating between those cases due to group A beta-hemolytic Streptococcus and those that are viral in origin.

Anti-DNA antibody derived from a systemic lupus erythematosus (SLE) patient forms histone-DNA-anti-DNA complexes that bind to rat glomeruli in vivo.

Histone can mediate the binding of both free DNA and DNA complexed to anti-DNA antibody to the glomerular capillary wall. We tested whether performed histone-DNA-anti-DNA immune complexes (IC) could bind to the glomerular capillary wall. The immune complex, generated with anti-DNA antibody derived from an SLE patient and excess of 125I-DNA followed by digestion with DNase, was mixed with histones. The complex containing 4 micrograms DNA was injected via the aorta into the left kidney of rats. At 15 min, 1-3% of the histone-DNA-anti-DNA antibody complex bound (measured as 125I-DNA), when histone was omitted less than 0-1% of the DNA-anti-DNA antibody complex bound. By immunofluorescence human immunoglobulins and histones, representing the IC, could be observed in a capillary pattern; but no complement deposition was detected. Electron microscopy revealed discrete, electron dense deposits in a subendothelial, subepithelial and mesangial localization at 15 min. These results provide direct evidence that antibodies from serum of SLE patients can form soluble histone-DNA-anti-DNA immune complexes that bind to the glomerular capillary wall in vivo.

Specificities and genetic characteristics of nucleosome-reactive antibodies from autoimmune mice.

Antinuclear antibodies are present in the serum of individuals with systemic autoimmune diseases such as SLE. Most autoantibodies characterized to date are directed against isolated nuclear molecules such as DNA or histones. We have obtained from spontaneously autoimmune mice six IgG mAb that recognize conformational nucleosome epitopes, but do not react with individual histones or DNA. For three of these mAb, the epitope is at least partially present in the H2A-H2B-DNA nucleosome subparticle, although their binding characteristics differ from those of conventional anti-H2A-H2B-DNA antibodies. All six mAb use VH or Vkappa genes which are recurrently utilized in anti-DNA and other antinuclear antibodies. The V regions of the nucleosome-reactive mAb also contain charged (mostly cationic) residues at sites that are likely to be critical for interaction with nucleosomal antigens. These results suggest that the usage of certain V gene segments in conjunction with suitable V(D)J rearrangements may confer reactivity to nucleosomal antigens. B cells producing such autoantibodies are probably expanded early during the autoimmune process. Somatic mutations in the V regions of nucleosome-reactive mAb may modulate their specificities and result in the acquisition of binding patterns restricted to individual chromatin components such as DNA.

Antigen-specific induction of antibodies against native mammalian DNA in nonautoimmune mice.

Spontaneous anti-DNA antibodies in autoimmune mice have the characteristics of antibody produced by Ag-specific, clonally selective B cell stimulation. The nature of the somatically derived antibody V region structures recurrent among spontaneous anti-DNA antibodies suggests that DNA or DNA-protein complexes may provide the antigenic stimulus for autoimmune anti-DNA antibody. In order to test this hypothesis directly, we have immunized normal, nonautoimmune-predisposed mice with complexes formed with DNA and an immunogenic, DNA-binding peptide. The highly immunogenic peptide, Fus1, forms an internal domain of a 128-amino acid ubiquitin-fusion protein from Trypanosoma cruzi. DNA-Fus1 complexes formed with native calf thymus DNA induced anti-DNA antibody in normal, nonautoimmune-predisposed mice that is similar in isotype and specificity to spontaneous anti-DNA antibody in (NZB x NZW)F1 autoimmune mice. The progressive nature of the development of dsDNA specificity in the immunized mice was also analogous to what is observed in the spontaneous anti-DNA antibody response of autoimmune (NZB X NZW)F1 mice. DNA-Fus1 immunized mice that produced IgG that bound to dsDNA had low to moderate levels of proteinuria and glomerular deposits of IgG. This experimental immunization system may be useful for understanding the immunologic basis for autoimmunity to DNA.

[Antibodies against deoxyribonucleoproteins in circulating immune complexes in the blood of patients with autoimmune diseases]. / Protilátky proti deoxyribonukleoproteinu v cirkulujících imunitních komplexech a sérech nemocných s nekterými autoimunitními chorobami.

In 40 patients with various autoimmune diseases antibodies against desoxyribonucleoprotein (DNP) were assessed by enzyme immunoanalysis (ELISA) in serum and after dissociation of the isolated precipitate of circulating immune complexes (CIC) in the presence of polyethylene glycol. The results indicate that antibodies against DNP are not specific for systemic lupus erythematosus (SLE) and can be detected in serum and in particular in CIC in various autoimmune conditions. In SLE they may be important for evaluation of the activity of the disease, in particular if estimated concurrently in serum and in CIC.

Immunogenic DNA-related factors. Nucleosomes spontaneously released from normal murine lymphoid cells stimulate proliferation and immunoglobulin synthesis of normal mouse lymphocytes.

The cell-free supernatants of normal spleen and thymus lymphocytes in short-term culture release low molecular weight (LMW) DNA protein molecules that have an immunoproliferative effect (polyclonal B cell activation) in vitro. We have determined that the protein-LMW DNA complexes responsible for these effects are nucleosomal constituents of chromatin, since the mitogenically active fractions of these cell-free supernatants contain the constituents of core histones (H3, H2A, H2B, H4) together with LMW DNA (140-180 bp), and since the immunoproliferative effects of these cell-free supernatants could be mimicked by various other nucleoprotein preparations (including calf thymus and chicken erythrocyte nucleosomes). The spontaneous cellular release of cleaved chromatin constituents in vitro can be attributed to a form of programmed cell death termed apoptosis, since the cultured spleen cells exhibited (a) morphologic evidence consistent with this process by electron microscopy, and (b) evidence of intracellular cleavage of chromatin which, like apoptosis, could be blocked with ZnSO4. This resulted in inhibition of the extracellular release of nucleosomal constituents as well as the immunoproliferative effects of the cell-free supernatants. The immunoproliferative effect of nucleosomes released from cells during apoptosis could be responsible for previously observed spontaneous in vitro anti-DNA and anti-histone antibody responses of murine spleen cells, and in vivo in normal lymphoid tissues, resulting in renewed cellular proliferation after cell death. In pathological states, this could result in abnormal polyclonal B cell proliferation and autoantibody formation.

Lupus erythematosus cell formation by a monoclonal antibody derived from an autoimmune MRL/Mp-lpr/lpr mouse.

An MRP-2 monoclonal antibody (MoAb) was primarily a product of hybridoma selected by binding to poly(ADP-ribose) from a lupus prone MRL/Mp-lpr/lpr (MRL/l) mouse, and was shown to cross-react with single-stranded (ss) DNA. Detailed examination revealed that MRP-2 MoAb bound to a conformational epitope formed between double-stranded (ds) DNA and total histone: both H3 and H4 were essential for the formation of this conformational epitope with dsDNA. Because of this characteristic of the MoAb, its ability to induce lupus erythematosus (LE) cells was examined in an indirect LE test with peripheral blood of MRL/Mp-+/+ (MRL/n) mice, which develop a mild form of lupus after the age of one year. MRP-2 MoAb was found to induce hematoxylin bodies, LE rosettes and LE cells, but a direct LE test using MRL/n mouse blood did not induce LE cell phenomena. This is the first demonstration of induction of LE cells by a MoAb that binds to dsDNA-histone complexes.

Characterization of antinuclear antibodies induced in rabbits by rinderpest virus infection.

Rabbits infected with the L strain of rinderpest virus (RV) produced high titres of antinuclear antibodies (ANA) which reached a maximum two weeks after inoculation but rapidly disappeared by 6-8 weeks. These ANAs reacted with HeLa cells by indirect immunofluorescence test resulting in a homogeneous nuclear fluorescence. In order to investigate the target antigens of ANAs, the effects on the nuclear fluorescence pattern of pretreating HeLa cells were examined: DNase 1 treatment resulted in a decrease in the fluorescence whereas no changes were evident after RNase A treatment. Some group of sera showed decreased fluorescence in the cells from which histones were acid extracted, but other groups did not change in fluorescence. Sera which had failed to react with acid extracted cells gave positive fluorescence following histone reconstitution. The results indicate that DNA and nucleohistone are the major target antigens for ANAs. In addition, antibodies against nucleoli and extractable nuclear antigens were induced in some rabbits.

Nuclear acceptor sites for sex steroid hormone receptors in chromatin.

The steroid receptor interactions in vitro with specific acceptor sites composed of acceptor protein-DNA complexes fulfill many of the criteria of a physiologically significant binding system. Chromatin acceptor sites for many steroid receptors (especially for the progesterone and estrogen receptor) are specific since they are saturable and competitive with unlabelled receptors, have high affinity for the receptor, distinguish between functional and nonfunctional receptors and demonstrate target tissue specificity. Pure DNA as acceptor sites does not display many of these properties. Therefore, it is clear that certain chromatin proteins provide the necessary specificity for the acceptor sites for the steroid receptors. For the progesterone receptor in the chick oviduct, these nuclear sites appear to contain specific chromosomal proteins as well as specific DNA sequences. The substitution of other chromosomal proteins or the genomic DNAs from evolutionarily distant organisms results in a loss of the specific nuclear binding. The nuclear acceptor sites appear to be resistant to the DNase activity which is not characteristic of transcriptionally active domains of the genome. Further studies using the ovalbumin gene sequences from genomic clones also indicate that none of the sequences within this domain and the 3-k flanking regions appear to contain the specific acceptor sequences. These observations have led to development of a model suggesting that the steroid receptors bind to acceptor sites distant from the structural genes the steroids ultimately regulate. Neighboring these acceptor sites are regulatory genes which code for regulatory substances which in turn (as secondary messengers) regulate at great distances the expression of the structural gene. This model might better fit the sex steroids which require 1-2 h to measurably alter gene transcription, as opposed to the glucocorticoids which more rapidly alter gene expression.

Characterization and antigenic specificity of chlorpromazine-induced antinuclear antibodies.

Prolonged treatment with chlorpromazine is often associated with the development of antinuclear antibodies, an immunoglobulin M lupus anticoagulant, and polyclonal serum IgM elevation, but not with clinical features of systemic lupus erythematosus (SLE). Sera from 62 long-term psychiatric patients given treatment daily with 100 mg or more of chlorpromazine for at least 1 year were screened for antinuclear antibodies by indirect immunoperoxidase assay using HEp-2 cells. In 26 samples, antinuclear antibody titers greater than or equal to 1:40 with a homogeneous pattern were seen when anti-human IgM was used as the second antibody, three sera samples reacted with IgG, and four samples reacted with both IgG and IgM antisera. The antinuclear antibody antigenic reactivity was investigated by using histone and nonhistone nuclear antigens by enzyme-linked immunosorbent assay and passive hemagglutination techniques. Forty serum samples reacted with histone. Twenty-five samples reacted with deoxyribonucleoprotein (DNP), 28 with single-stranded DNA, and two with double-stranded DNA. No reaction was obtained with the extractable nuclear antigens RNP or Sm. These results indicate that chlorpromazine-induced antinuclear antibodies, like the antinuclear antibodies induced by hydralazine and procainamide, react mainly with histone nuclear antigens. Unlike the hydralazine and procainamide response, in which both IgG and IgM antibodies are demonstrated, the chlorpromazine-induced autoantibodies are predominantly of the IgM class.

Autoantibodies and their relation to rheumatic diseases.

The use of tissue culture substrates for immunofluorescence determinations of nuclear, cytoplasmic, and mitotic cell-related autoantibodies has resulted in the delineation of diverse new specificities, whose clinical correlates are now becoming apparent. This review details both major and minor autoantibody specificities, the status of knowledge regarding their target antigens, and the relation of these serologic systems to distinctive rheumatic disease syndromes.

Antinuclear antibodies as indicators for the procainamide-induced systemic lupus erythematosus-like syndrome and its clinical presentations.

Fifty patients on a regimen of procainamide were studied in regard to the association between antinuclear antibodies (ANA) and the development of drug-induced systemic lupus erythematosus (SLE)-like syndrome. Four groups were identified: Group 1 was ANA-positive, with clinical manifestations (serologic and clinical findings); Group 2 was ANA-positive, without clinical manifestations (serologic findings only); Group 3 was ANA negative (no patients with clinical manifestations); and Group 4 had SLE persisting after discontinuance of procainamide. The leukocyte-specific ANA (LSANA) patterns were predominant, with peripheral LSANA confined to Groups 1 and 4. Furthermore, the titer of the homogeneous LSANA, to which peripheral LSANA converts on dilution, was clinically significant. A homogeneous LSANA titer of 160 or greater was seen essentially only in patients with clinical manifestations of the SLE-like syndrome. Serial ANA determinations are therefore necessary to monitor patients receiving procainamide.

[Physiological significance of specific immunoglobulins in regulating steroidogenesis in adrenal cortex cells]. / Fiziologicheskoe znachenie spetsificheskikh immunoglobulinov v reguliatsii steroidogeneza v kletkakh korkovogo veshchestva nadpochechnikov.

The data obtained suggest the existence of physiological immunoglobulin mechanisms regulating the functions of the cells including hormone--producing ones. Serologically identical antigens were revealed both in the blood and the extracts from adrenocortical cell nuclei of intact rats. Apart from that, autoantibodies to deoxyribonucleoprotein (DNP) of adrenocortical cells were found in the blood sera of normal animals. Immunoglobulins G (ig G) against adrenocortical cell nuclei, DNP and the nuclei partially devoid of DNP, were produced. The specific IgG were shown to stimulate the steroidogenesis in target cells. The penetration of the specific antibodies into the target cell nuclei in vivo was proved. The data obtained support the hypothesis of the nuclear mechanisms of antibody--caused cytostimulation.