Localization of fluorescein-labeled antinucleoside antibodies in glomeruli of patients with active systemic lupus erythematosus nephritis
01 Nov 1970-Journal of Clinical Investigation (American Society for Clinical Investigation)-Vol. 49, Iss: 11, pp 2106-2118
TL;DR: The findings are consistent with the hypothesis that antigen-antibody complexes, formed by denatured DNA, specific antibody, and complement, are present in the deposits of foreign material accumulated in the GCW of patients with active SLE glomerulonephritis, and that they may contribute to the pathogenesis of this renal disease.
Abstract: Renal tissues from two groups of patients were studied with fluorescein-labeled (Fl-) antibodies (Abs) to immunoglobulins, complement, and antibodies prepared in rabbits against BSA conjugate of 5-methyluridine (T) and cytidine (C), the latter two of which react specifically with denatured DNA. The first group consisted of 13 SLE patients, and the second consisted of 53 patients with non-SLE nephropathies. The data obtained from the two groups of patients were used for comparison, and they showed the following:(a) Fl-Abs to immunoglobulins and complement were bound in the glomeruli of tissues from all patients with active SLE glomerulonephritis characterized by deposits of foreign material in glomerular capillary walls (GCW). The fluorescent pattern was granular, corresponding to the distribution of the glomerular deposits, as seen by electron microscopy. Fl-Abs reactive with thymine and cytosine were bound in the GCW of eight of the nine patients with active SLE glomerulonephritis and showed the same granular distribution. The capacity of these latter Fl-Abs to stain the GCW was removed by absorption with the homologous antigen or denatured DNA.(b) Fl-Abs to immunoglobulins, complement, and pyrimidine bases of DNA did not react with the GCW of two SLE patients without clinical and histologic evidence of glomerulonephritis or with the sclerotic glomeruli of two uremic patients with chronic "burned out" lupus nephritis.(c) The glomeruli of 47 of the 53 patients with other nephropathies bound Fl-Abs to immunoglobulins and complement to some extent, and in 26, the localization appeared as marked as in the patients with active SLE glomerulonephritis. Fl-Abs reactive with thymine and cytosine were bound in the GCW of only one of the renal tissues from the 53 non-SLE patients. In the remaining 52, no binding was seen.(d) The findings are consistent with the hypothesis that antigen-antibody complexes, formed by denatured DNA, specific antibody, and complement, are present in the deposits of foreign material accumulated in the GCW of patients with active SLE glomerulonephritis, and that they may contribute to the pathogenesis of this renal disease.
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TL;DR: Autoantibodies to nuclear antigens (ANAs) have assumed an important place in the diagnostic armamentarium of the clinician because of distinct profiles of ANAs in different diseases.
Abstract: Publisher Summary Autoantibodies to nuclear antigens (ANAs) have assumed an important place in the diagnostic armamentarium of the clinician because of distinct profiles of ANAs in different diseases. Profiles of ANAs have, therefore, been extremely useful in differential diagnosis, where the disease does not have classical or full-blown manifestations.. Immune complexes formed in the circulation or in situ mediate tissue injury by the activation of complement and other inflammatory mediators. Not only do these antibodies precipitate their respective antigens but also other proteins or nuclear RNAs that might be associated with them in special ways. The reasons for these special associations of protein antigens with specific sets of nuclear RNAs is unknown, but the possibility that there might be functional relationships in these complexed particles is not unreasonable. The key question that pervades the minds of many investigators in this field is the reason for the appearance of ANAs in certain individuals. It is highly improbable that the phenomenon is a random immune response to nuclear breakdown products, because the types of ANAs in different diseases are strikingly different. Some known environmental agents are drugs, such as hydralazine and procainamide, that together with lower levels of hepatic acetyltransferase enzyme predispose the host to the development of ANAs. Another agent may be the Epstein–Barr virus that is a ubiquitous environmental agent.
881 citations
TL;DR: This chapter summarizes the data presented in two reviews of experimental acute and chronic immune complex disease produced by nonliving antigens and discusses in detail more recent studies.
Abstract: Publisher Summary No experimental model has provided greater insight into the mechanism of immune complex disease than the experimental serum sickness. The morphological, immunohistological, and serological features of the laboratory models have provided a basis for understanding the pathogenic mechanisms responsible for human glomerulonephritis, vasculitis, and a variety of systemic connective tissue diseases. The subject of experimental acute and chronic immune complex disease produced by nonliving antigens has received extensive review in this series. This chapter summarizes the data presented in these two reviews and discusses in detail more recent studies. Experiments to study acute immune complex disease (serum sickness) have been performed in rabbits almost exclusively. Experimental chronic immune complex disease has proved to be a most useful model in understanding human glomerulonephritis. When injected daily with heterologous serum protein antigens, rabbits with strong antibody responses develop chronic membranous glomerulonephritis in about 5 weeks.
539 citations
TL;DR: This chapter describes the interaction of immune complexes (ICs) with complement and with the cells of the immune system, thereby making it possible to identify the antigens involved in immune processes of a great many diseases, including those of unknown etiology.
Abstract: Publisher Summary This chapter describes the interaction of immune complexes (ICs) with complement and with the cells of the immune system. The effect of immune complexes depends to a great extent on their antigen–antibody ratio so that their influence, either stimulatory or suppressive, is itself modulated by quantitative aspects of the immune response. The development of numerous techniques for the detection and quantitation of immune complexes has stimulated clinically related research and expanded the list of diseases in which immune complexes appear to play an important role. An extension of this diagnostic technology is the ability to isolate immune complexes and, in turn, their antigenic component, thereby making it possible to identify the antigens involved in immune processes of a great many diseases, including those of unknown etiology. In vivo and in vitro experiments have clarified many factors involved in IC formation, removal, and localization as well as the mechanisms of IC-induced inflammatory reactions. An individual can make an immune response to a large number of exogenous and a smaller number of endogenous antigens. Depending upon the availability of antigen, the antibodies so produced form ICs, which for the most part serve the purpose of aiding the host in eliminating potential pathogens.
434 citations
TL;DR: A major role for high avidity antinative-DNA in DNA/antiDNA immune complex-induced glomerular injury in systemic lupus erythematosus is supprot.
Abstract: Significant differences in both specificity and avidity of anti-DNA antibodies were observed in the sera of groups of patients with active systemic lupus erythematosus glomerulonephritis, active systemic lupus erythematosus without nephritis, and in IgG eluates obtained by DNAase digestion of isolated glomeruli from glomerulonephritic kidneys. With methylated albumin-kieselguhr fractionated 3H-HeLa DNA as a source of native or single-strand DNA antigen in a modified Farr assay, an increased level of antibody to native DNA was associated with active systemic lupus erythematosus, particularly active nephritis. The avidity of antinative DNA estimated from plots of the reciprocals of bound and free antigen according to the Sips distribution formula was significanly lower in active glomerulonephritis sera than in sera from patients with active systemic lupus erythematosus without nephritis. However, antinative DNA of uniformly high avidity was found in the glomerular eluates. Avidity of single-strand DNA antibodies did not differ in the various patient groups. The data stronly supprot a major role for high avidity antinative-DNA in DNA/antiDNA immune complex-induced glomerular injury in systemic lupus erythematosus.
403 citations
TL;DR: CIE is a rapid and sensitive technique for detecting precipitating antibodies to a number of nuclear acidic antigens and methods are described to identify the immunochemical specificities of the precipitin lines by the use of standard reference sera.
Abstract: Saline extracts of rabbit thymus were found to contain many nuclear antigens that reacted with antibodies in the sera of patients with systemic rheumatic diseases. Counterimmunoelectrophoresis (CIE) was used to detect antibodies to nuclear acidic protein (Sm), nuclear ribonucleoprotein (RNP), and antibody to nuclear antigen B, which was reported previously in Sjogren's syndrome. All these nuclear antigens behaved as anions with different mobilities in CIE and could be distinguished from one another by the locations of the precipitin lines. They could also be distinguished by the facts that the nuclear RNP precipitin lines were abolished by digestion with ribonuclease whereas others were unaffected, and that Sm precipitin lines developed later than other precipitin lines. With this technique antibody to nuclear RNP was detected in 46% of patients with systemic lupus erythematosus (SLE) compared to 26% detected by the hemagglutination technique. Similarly the increased sensitivity of the CIE technique was able to show that antibody to B antigen was present in 12% of SLE patients, whereas this antibody was not detectable in the same group of patients by immunodiffusion. This study shows that CIE is a rapid and sensitive technique for detecting precipitating antibodies to a number of nuclear acidic antigens. Methods are described to identify the immunochemical specificities of the precipitin lines by the use of standard reference sera.
359 citations
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TL;DR: The immunochemical evidence for the high specific activity of antinuclear antibodies and the association of DNA antigen with DNA antibody in glomeruli add further support for the antigen-antibody complex hypothesis for renal injury in systemic lupus erythematosus.
Abstract: Antibodies were eluted from the isolated glomeruli prepared from the kidneys of 10 patients with the nephritis of systemic lupus erythematosus. Antibodies reacting primarily with buffer extracts of nuclei were eluted by acid treatment, and antibodies reacting mainly with DNA and nucleoprotein were eluted with deoxyribonuclease. Quantitative immunochemical studies revealed a high concentration of antinuclear antibody per milligram of γ-globulin in glomerular eluates compared with that in the corresponding serums. The γ-globulin of two eluates was found to consist predominantly of antinucleoprotein antibody. The selective elution of antinuclear antibodies was also indicated by the absence of other serum antibodies in the eluates. DNA antigen was demonstrated in the glomeruli of two kidneys with nephritis by means of isolated anti-DNA antibody labeled with fluorescein. In one of these cases, anti-DNA antibodies were also found concentrated in the glomeruli and, in the second, circulating anti-DNA antibodies were demonstrated in the patient's serum. The immunochemical evidence for the high specific activity of antinuclear antibodies and the association of DNA antigen with DNA antibody in glomeruli add further support for the antigen-antibody complex hypothesis for renal injury in systemic lupus erythematosus.
797 citations
772 citations
TL;DR: These findings show that the major component of bacteriochlorophyll is inert with respect to the foregoing light-induced activities, and that a special kind of reaction center is needed for the photochemistry that leads to photosynthesis.
Abstract: c or reduced phenazine methosulfate coupled with the reduction of ubiquinone. We wish to report now that chromatophores from aerobically grown R. spheroides, strain Ga, sensitize these reactions efficiently (with quantum requirements of a few quanta per electron transfer), whereas chromatophores from the nonphotosynthetic mutant strain PM-8 are entirely unable to drive these photochemical processes. The failure of strain PM-8 to catalyze the photooxidation of reduced phenazine methosulfate is the more remarkable because this reaction is sensitized by purified bacteriochlorophyll in vitro. These findings show that the major component of bacteriochlorophyll is inert with respect to the foregoing light-induced activities, and that a special kind of reaction center is needed for the photochemistry that leads to photosynthesis. The results of experiments with exogenous reagents will be published in detail elsewhere.
433 citations
TL;DR: Sera from patients with active lupus erythematosus fixed complement with a wide variety of nuclei from different organs and species, with calf thymus nucleoprotein, and in two instances with histone, suggesting the presence of 2 distinct serum factors.
Abstract: Summary1. Sera from patients with active lupus erythematosus fixed complement with a wide variety of nuclei from different organs and species, with calf thymus nucleoprotein, and in two instances with histone. Isolated calf thymus, salmon sperm, human leukocyte and pneumococcal DNA also fixed complement with many of these sera. Similar reactions were not encountered in a limited control series including normal individuals and other pathological states. 2. Most active L.E. sera fixed complement with both nuclei and DNA in roughly parallel titer. However, exceptions were encountered and one serum reacted strongly with nuclei but failed to react with DNA. Cross-absorption experiments with nuclei and DNA suggested the presence of 2 distinct serum factors. 3. The L.E. factor appeared to be related to the factor responsible for complement fixation with nuclei but distinct from that responsible for DNA fixation. 4. The significance of these findings with respect to antibodies against nuclear constituents is disc...
266 citations
TL;DR: Renal biopsies from 16 patients with nephrosis, 7 patients with glomerulonephritis, and 3 patients with disseminated lupus erythematosus were studied with the electron microscope to indicate that early in the course of each of these diseases alterations occur in the fine structure of the glomeruli which serve to distinguish one disease process from another.
Abstract: Renal biopsies from 16 patients with nephrosis, 7 patients with glomerulonephritis, and 3 patients with disseminated lupus erythematosus were studied with the electron microscope. The observations presented indicate that early in the course of each of these diseases alterations occur in the fine structure of the glomeruli which serve to distinguish one disease process from another. In nephrosis, some distortion of the organization of the epithelial foot processes was seen in all patients. These epithelial changes constituted the early, consistent lesion of the disease. There was frequently also a swelling of the endothelium. In glomerulonephritis, pronounced proliferative changes involving the endothelium and to a lesser extent the epithelium, together with the laying down of a basement membrane-like material, represented the predominate pathologic processes. There was also a swelling of both endothelial and epithelial cytoplasm. The epithelial foot processes generally appeared normal. In patients with a clinically "mixed" picture of nephrosis and nephritis, the glomerular changes were likewise "mixed," for various combinations of epithelial, endothelial, and basement membrane abnormalities were present. In disseminated lupus erythematosus, a more or less generalized thickening of the basement membrane proper associated with a variable degree of endothelial proliferation was seen. It is suggested that an accentuation of the process of basement membrane thickening results in the "wire loop" appearance sometimes seen by light microscopy. Although the earliest alterations in glomerular fine structure were characteristic for each of the disease processes, at later stages the changes were not always distinctive. The resulting scarred or "hyalinized" glomeruli, composed of relatively homogeneous, basement membrane-like material, and a few atrophic cells, appeared quite similar. Although the functional implications of the structural changes observed remain obscure at this time, it is believed that insight into mechanisms may stem from such observations.
256 citations