Article

Gamma G globulin subgroup composition of the glomerular deposit in human renal diseases

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Abstract

36 renal biopsies from patients with nephritis were studied for glomerular localization of the heavy chain subgroups of immunoglobulin G (IgG or gammaG). The deposition pattern of these subgroups was selective and did not reflect the normal serum concentration of these proteins. gammaG2, which comprises 18% of normal serum gammaG, was the predominant or unique subgroup deposited in five cases of lupus nephritis and four biopsies with other forms of nephritis associated with granular gammaG deposits. gammaG3, which normally makes up only 8% of the serum gammaG, was the dominant subgroup seen in one biopsy of lobular glomerulonephritis. Patients with linear gammaG deposits generally had a selective absence of gammaG3 and often had large amounts of gammaG4 (normally 3% of the serum gammaG) deposited. The deposition of complement components C1q, C4, and C3 was variable. One biopsy had only gammaG2 and no complement components in the deposits and had no neutrophile leukocyte infiltration. This latter observation correlates well with the poor ability of gammaG2 to fix complement in vitro. Similarly, deposits containing large amounts of gammaG4, which does not fix complement, also tended to have less inflammatory infiltrate than deposits devoid of this subgroup. The selective deposition of monotypic or restricted gammaG subgroups on the glomerulus supports the likelihood that the gammaG represents antibody. The nature of the subgroup involved in the deposit may represent one variable in the determination of the inflammatory and morphological picture that evolves in human glomerulonephritis.

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Article
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Studies were performed to determine if cultured human endothelial cells synthesized basement membrane collagen. In culture, endothelial cells were attached to grossly visible membranous structures which on light microscopy were composed of ribbons of dense, amorphous material. On transmission electron microscopy, these membranous structures consisted of amorphous basement membrane, and material morphologically similar to microfibrils and elastic fibers. By immunofluorescence microscopy, these membranous structures stained brightly with antisera to human glomerular basement membrane. Cultured endothelial cells incorporated [3H]proline into protein; 18% of the incorporated [3H]proline was solubilized by purified collagenase. When endothelial cells were cultured with [14C]proline, 7.1% of the incorporated counts were present as [14C]hydroxyproline. Cultured endothelial cells were labeled with [3H]glycine and [3H]proline and digested with pepsin. The resulting fractions on analysis by SDS-polyacrylamide gel electrophoresis contained two radioactive protein peaks of mol wt 94,200 and 120,500. Both these peaks disappeared after digestion with purified collagenase. The peak of mol wt 120,500 corresponds to that of alpha1 (IV) collagen; the peak of the mol wt 94,200 probably corresponds to that of alpha1 (III) collagen. Thus, cultured human endothelial cells synthesize material which is morphologically and immunologically like amorphous basement membrane and biochemically like basement membrane collagen. Cultured endothelial cells probably also synthesize material which is morphologically similar to microfibrils and elastic fibers.
Article
Anti–glomerular basement membrane antibody-induced glomerulonephritis. Sixty-three glomerulonephritic patients with evidence of anti–glomerular basement membrane (GBM) antibody by immunofluorescence, elution of specific antibody and/or detection of circulating antibody are reviewed. Thirty-two of the patients had concomitant pulmonary manifestations (hemorrhage and/or pulmonary failure) compatible with Goodpasture's syndrome, 21 had only glomerulonephritis, and ten without prior immunopathologic studies had antiGBM antibodies in renal transplants and no history of pulmonary symptoms. The majority of the patients were young males who succumbed to the disease process or required dialysis and/or transplantation. Five males and two females did not develop permanent renal insufficiency. Bilateral nephrectomy (19 patients) was felt to be helpful or life-saving in seven patients; however, six patients had continued pulmonary symptoms after nephrectomy, contributing to death in three cases. Five of 13 nonnephrectomized or unilaterally nephrectomized individuals died in pulmonary failure. Immunosuppressive therapy was not generally helpful. Circulating antiGBM antibodies persisted for an average of six to eight months after bilateral nephrectomy. Transplantation was usually successful when delayed until circulating antiGBM antibodies had declined; however, evidence of recurrent glomerulonephritis developed in 19, possibly 20, of 34 transplanted patients, resulting in graft failure in seven, possibly eight, instances.
Article
Absence of C2 was identified in an 18-year-old white male with progressive and rapidly fatal lupus nephritis. Genetic studies of the patient and 8 family members linked this deficiency with the HLA haplotype A10/B18, for which the patient was homozygous. High titers (1:1600) of anti-RNP antibodies, as well as antibodies to double-stranded DNA, were present. Serum levels of properdin factor B were persistently decreased, whereas levels of C1q and C3 intermittently were low. Biopsies of skin and kidney showed typical SLE histopathology with deposition of immunoglobulins and early- and late-acting C components; properdin was deposited in the kidney. Despite the inability of the patient's serum to mediate lysis through the classic C pathway, C-dependent lysis through the alternative pathway was readily achieved using either unsensitized rabbit cells in gels or mixtures of guinea pig cells and zymosan as the indicator system. These studies thus reveal a new association of fatal lupus nephritis with homozygous C2 deficiency and show the usefulness of two new assays for measuring the hemolytic capacity of the alternative C pathway in human serum. The decreased levels of properdin factor B in serum, deposition of properdin and C3 in renal tissue, and capacity of C2-deficient serum to mediate hemolytic C activity through the properdin pathway in vitro implicate the alternative C pathway in the severe tissue damage observed.
Chapter
Renal symptoms can accompany infectious diseases in two periods: (i) at the time of the onset of the infection, or (ii) after a latent period corresponding to the time needed for the production of human antibodies. In the first case we talk about early, intrainfectious or para-infectious nephritis, and in the latter, about postinfectious glomerulonephritis. Early nephritis is caused with all certainty by primary toxic effects, while in the development of postinfectious glomerulonephritis the antigen-antibody reaction plays the decisive role. It was realized as early as the beginning of this century that scarlatinal nephritis is an allergic disease [265]. Since then a number of explanations have been put forward for the aetiology and pathomechanism of post-streptococcal glomerulonephritis including the theory of autoimmunization. Nevertheless, no final solution of the problem has been reached. Some authors are of the opinion that autoimmunization is not the cause but the consequence of renal disease.
Article
In an attempt to document progression rate differences in the development of glomerular lesions in mink infected with Aleutian disease virus (ADV), the glomeruli of Aleutian and non-Aleutian mink experimentally infected with ADV were evaluated by light, fluorescent, and electron microscopy. The animals were also examined for the presence of interstitial infiltrate, neutrophils, and arterial lesions. One hundred percent of the Aleutian mink had glomerular cell proliferation and interstitial infiltrate, while 95% of the Aleutian and 41% of the non-Aleutian mink had neutrophilic infiltrates and arteritis, respectively. Of the non-Aleutian mink, 91, 83, 42, and 12.5% had glomerular cell proliferations, glomerular neutrophils, interstitial infiltrate, and arterial lesions in, that order. All the Aleutian mink had glomerular depositions of gamma-globulin (IgG) and complement (C3), whereas 75% of non-Aleutian mink had deposits of IgG and C3. One hundred percent of both genotypes had glomerular deposits of immunoglobulin M (IgM). Ultrastructural glomerular changes consisting primarily of depositions of granular electron-dense material on basement membranes were observed in Aleutian mink 6 weeks after infection and 12 weeks after infection in non-Aleutian mink. These findings document progression rate differences in the development of glomerular lesions in Aleutian disease-affected Aleutian and non-Aleutian mink. Further, they emphasize the need for exploration of pathogenetic mechanisms involved in progression rate differences in lesion development.
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Studies were performed to determine if cultured human endothelial cells synthesized basement membrane collagen. In culture, endothelial cells were attached to grossly visible membranous structures which on light microscopy were composed of ribbons of dense, amorphous material. On transmission electron microscopy, these membranous structures consisted of amorphous basement membrane, and material morphologically similar to microfibrils and elastic fibers. By immunofluorescence microscopy, these membranous structures stained brightly with antisera to human glomerular basement membrane. Cultured endothelial cells incorporated [3H]proline into protein; 18% of the incorporated [3H]proline was solubilized by purified collagenase. When endothelial cells were cultured with [14C]proline, 7.1% of the incorporated counts were present as [14C]hydroxyproline. Cultured endothelial cells were labeled with [3H]glycine and [3H]proline and digested with pepsin. The resulting fractions on analysis by SDS-polyacrylamide gel electrophoresis contained two radioactive protein peaks of mol wt 94,200 and 120,500. Both these peaks disappeared after digestion with purified collagenase. The peak of mol wt 120,500 corresponds to that of alpha1 (IV) collagen; the peak of the mol wt 94,200 probably corresponds to that of alpha1 (III) collagen. Thus, cultured human endothelial cells synthesize material which is morphologically and immunologically like amorphous basement membrane and biochemically like basement membrane collagen. Cultured endothelial cells probably also synthesize material which is morphologically similar to microfibrils and elastic fibers.
Article
Lymphocytes with surface IgG and plasma cells producing IgG have been shown to be a prominent feature of different inflammatory periodontal lesions. The subclass nature of the IgG-bearing cells in gingival biopsies was investigated in order to further define these inflammatory cell infiltrates and correlate them with different stages of human periodontal disease.Mild gingivitis was characterized by lymphocytes which lacked surface IgG and Fc receptors suggestive of thymus (T) dependent cells. In severe gingivitis, the nature of the localized cellular infiltrate changed with increasing numbers of IgG1 (22%) and IgG3 (17%) labelled lymphocytes, with lesser number of IgG4 (7%) and IgG2 (1%) similar incidence of IgG subclass plasma cells was also observed. Destructive periodontitis was characterized by high numbers of plasma cells (57%) distributed throughout the gingiva. The incidence of IgG subclasses was 25% IgG1, 19% IgG4, 18% IgG3, and 1% IgG2. This distribution of IgG subclass. These findings present further evidence that the cellular infiltrate in the most destructive stage of periodontal disease, priodontitis, is primarily composed of bone marrow (B) derived cells.
Article
Systemic lupus erythematosus (SLE) was documented in a patient with genetic deficiency of the second complement component (C2). A review of disease manifestations in this patient and in others with SLE and genetic C2 deficiency previously reported on suggest that many findings such as fever, skin lesions, central nervous system involvement, the presence of autoantibodies and leukopenia occur in patients with SLE independent of whether or not they have C2 deficiency. However, renal disease appears mild in patients with genetic C2 deficiency despite the presence of immunoglobulins and complement components demonstrated in glomeruli by immunofluorescent microscopy. Electron microscopic study of glomeruli from the patient we describe showed electron-dense deposits consistent with the immunofluorescent findings. Tubuloreticular inclusion bodies identical to those previously seen in patients with SLE were also observed. Analysis of serum complement components and tissue deposition of complement components suggests activation of both the classic and alternative complement pathways in patients with C2 deficiency. The clinical, pathologic and complement findings in this group of patients support the hypothesis that although SLE is similar in patients with and without C2 deficiency, renal disease remains mild when this classic pathway component is not present. Histocompatibility antigen analysis of the family of the propositus confirmed the association of C2 deficiency with an A10, B18 haplotype, and with a haplotype not previously associated with C2 deficiency, AW32, BW40. One sibling with this latter haplotype has normal C2 levels, presumably as a result of recombination.
Article
Although an unusually high incidence of immunological diseases has been described in patients with hereditary C2 deficiency, the severity of these illnesses has been relatively mild, suggesting that blocking complement activation beyond C4 may protect against significant complement-mediated inflammation. This report describes studies in a C2-deficient patient with severe systemic lupus erythematosus (SLE) and diffuse proliferative glomerulonephritis. An immunopathological study of the kidney revealed the deposition of properdin, properdin factor B, C3 and C5 in a pattern similar to immunoglobulin G deposits. Serum properdin and properdin factor B levels were low at various times during the patient's course. In vitro complement fixation studies showed C3 fixation by glomerular deposits could occur via the alternative pathway. Studies of the immune deposits in the patients' skin revealed similar results. These studies suggest that inflammation may be effectively mediated via the alternative complement pathway in the C2 deficiency-lupus syndrome.
Article
The in vivo complement deposition and in vitro complement reactivity of immune aggregates were studied in the lungs of five patients with systemic lupus erythematosus (SLE). By direct and indirect immunofluorescence, C1q deposition appeared comparable to Ig deposition, but deposition of other complement proteins (C4, C2, C3c, C5, B, P) was scant or absent. In contrast, glomerular and renal vascular immune deposits in these patients showed extensive localization of these complement components. The pulmonary immune aggregates were capable of in vitro complement fixation of C4, C3c, and C5. These studies suggest that complement deposition may be limited at sites of pulmonary immune aggregation in certain patients with SLE, and that factors other than intrinsic complement reactivity appear to be responsible for the observed complement component composition of immune deposits in the lungs of these patients.
Article
The originally dismal prognosis associated with anti-GBM Ab-mediated GN and Goodpasture's syndrome may be changing as we recognize a broader spectrum of disease, improve general supportive care, and improve specific treatment. Immunosuppressive therapy, if started early in the course of disease, may prevent or allow recovery from renal failure and may also result in cessation of recurrent pulmonary hemorrhage in most patients with this form of Goodpasture's syndrome. The administration of pharmacologic doses of corticosteroids intravenously can result in cessation of and dramatic recovery from severe pulmonary hemorrhage and obviate the need for emergency bilateral nephrectomy. Plasmaspheresis may represent a useful therapeutic procedure for the immediate and long term reduction in amounts of circulating anti-GBM Ab, but the definition of its true value and role awaits completion of controlled, prospective trials. Immunosuppressive therapy, with or without plasmapheresis, can reduce quantities of anti-GBM Ab in serum to undetectable levels without nephrectomy. Thus, it is likely, but not proven, that nephrectomy can be discontinued as a routine pretransplantation procedure in patients with anti-GBM Ab mediated GN. Finally, in patients who suffer irreversible renal failure, renal transplantation can be successfully undertaken with minimal risk of recurrent disease, when circulating anti-GBM Ab becomes undetectable.
Article
Nonimmunoglobulin C3 activating factor in membranoproliferative glomerulonephritis. Material isolated by cryoprecipitation from the serum of a patient with membranoproliferative glomerulonephritis exhibited alternative complement pathway C3-splitting activity associated with a nonimmunoglobulin γ-migrating protein. The cryoprecipitate required factor B and magnesium ions, but not C1, C2, or calcium ions for activity. Antiserum produced against the cryoprecipitate recognized a previously unidentified γ-migrating material, as well as β1H globulin (C3b inactivator-accelerator), and κ light chains. Immunoglobulin (Ig) G was present in trace quantities. Immunoadsorption with antisera to human γ heavy chains or κ light chains did not alter C3 activation by the resolubilized material. The γ-migrating material, which appears to be an altered form of a normal α2 protein, retains its C3 splitting property when separated from β1H and IgG by sucrose density ultracentrifugation. Antisera monospecific to β1H and to the γ-migrating material each produced intense glomerular immunofluorescence in the patient's renal biopsy in a pattern similar to that of anti-C3. No glomerular staining occurred with antihuman IgG, M, A, E, Clq, C4, or C2. It is proposed that interaction between this protein and β1H in the circulation and in the glomerulus could block β1H-dependent decay-dissociation of C3 convertases, resulting in unregulated alternative complement pathway C3 activation and complementmediated glomerular damage.
The subclasses of IgG antibodies formed by grass pollen-allergic patients during immunotherapy were investigated by using a radioallergosorbent test (RAST) and a quantitative immunofluorescence method known as the defined antigen substrate spheres (DASS) system. By the use of rabbit antisera directed against the subclasses of IgG, the specificity of which was checked in the passive hemagglutination and immunofluorescence techniques, it was shown that a relatively high proportion of the grass pollen-specific antibodies belonged to the IgG4 subclass. Apart from the high binding activity of IgG4 which increased during treatment, a moderate binding activity of the other subclasses was also found. Binding of all subclasses increased slightly in the pollen season and could be specifically blocked by perincubation with soluble grass pollen extract. The results of the IgG4 binding, determined in vitro with the DASS system, and the blocking activity of the sera, determined in vivo by skin tests are suggestive for a relation between these activities. Also in the group of patients with a low IgE-RAST score, the skin reactivity decreased as the IgG4 binding activity increased.
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The composition of various isolated antibodies was determined by quantitative analyses for heavy chain subgroups and light chain types. Certain antibodies such as anti-tetanus toxoid and anti-A isoagglutinins were predominantly of the major gammaG1-type. However, a high preponderance of molecules of the minor gammaG2-subgroup was found for antibodies to dextran, levan, and teichoic acid. These findings explain some unusual features previously noted for anti-dextrans such as weak PCA reactions and lack of Gm antigens. Studies of several isolated antibodies from single heterozygous individuals showed a selective absence of genetic markers in certain antibodies and their presence in others. The "allelic exclusion" principle was clearly evident in the isolated antibodies of two different individuals. Large differences in the ratio of kappa to lambda light chains were observed for the same type of antibody from different individuals. Subfractionation of dextran antibodies by affinity for specific glycosidic linkage or combining site size produced marked changes in the ratios. The isomaltohexaose eluates of the dextran antibodies from two subjects were primarily kappa and the isomaltotriose eluates were predominantly lambda. The one anti-levan antibody studied was uniquely homogeneous, consisting exclusively of gammaG2-heavy chains and kappa light chains. By these criteria as well as others, it closely resembled myeloma proteins.
Article
Daily injections of any one of several foreign serum proteins produced in rabbits functional and morphological alterations similar to those seen in acute, subacute, and chronic human glomerulonephritis. The critical factor determining whether a rabbit would develop renal disease and the type of disease developed was the amount of antibody the rabbit formed. Those responding with much antibody were likely to develop an acute, self-limited glomerulonephritis and to be subsequently immune to further renal damage. Those responding with antibody barely sufficient to neutralize the antigen injected developed subacute and chronic glomerulonephritis. In the circulation of the rabbits with chronic glomerulonephritis, there was a daily recurring antigen-antibody reaction in the region of near antigen excess to near antibody excess which presumably led to the disease. Antigen apparently in the form of antigen-antibody complexes was deposited along the renal capillary basement membranes coincident with the development of subacute and chronic glomerulonephritis. Once developed, the morphologic stigmata of chronic glomerulonephritis persisted even after injections of antigen were stopped. However, in milder instances the renal function recovered in part after stopping antigen. This experimental model has several implications: first, the renal injury is precipitated by antigens with no known affinity for, or immunologic relationship to, kidney; second, antigen antibody complexes localize in the kidney, apparently on the basis of non-immunologic factors, and may be an etiologic agent of renal injury; third, severe hypersensitivity disorders can be related specifically to relatively poor as well as to good antibody responses; and finally, the pathogenesis suggested here offers an alternative to that of nephrotoxic serum nephritis for the experimental approach to the study of human glomerulonephritis.
Article
The primary phase of nephrotoxic serum nephritis produced by rabbit nephrotoxic serum appears to be dependent to a great extent, but not completely, upon the participation of serum complement. On the other hand, duck nephrotoxic serum produces its primary renal injury without detectable utilization of or dependence upon serum complement. The secondary phase of nephrotoxic serum nephritis appears to be largely or entirely dependent upon the host's antibody response to the heterologous gamma globulin fixed in the glomeruli. No evidence could be obtained for the existence of an autoimmune antikidney response by the host in this experimental model.
Article
Excerpt More than twenty known genetic antigens have been delineated on the heavy chains of human γ globulin (Martensson, 1966; Natvig and Kunkel, 1967a). These are distributed among three different subgroups of γG globulin and have served as genetic markers for the elucidation of the relationship between the different structural genes that determine heavy chain synthesis (Kunkel, Allen, and Grey, 1964; Terry, Fahey, and Steinberg, 1965). Certain heavy chains have multiple genetic antigens in different areas which can be separated by papain splitting. Information concerning the genetic control of different portions of these chains can thus be obtained. Two new genetic antigens, Gm (n) and Gm (g), have proven of particular utility in understanding the differences between the subgroups (Kunkel, Yount, and Litwin, 1966; Natvig, 1966). Varying relationships between genetic markers in different populations and within certain rare families have offered evidence for recombination between the different cistrons involved. These studies...
Article
These observations established the presence of anti-GBM antibodies in the sera and/or kidneys of six humans with glomerulonephritis. Further, it seems clear that these antibodies do combine with the host's glomeruli in vivo and with GBM antigen of several species in vitro. Transfer of acute glomerulonephritis to normal recipient monkeys was possible with serum or renal eluate IGG from the three patients with anti-GBM antibodies in whom sufficient material was available. Based on this transfer of nephritis and on the presence of these antibodies at the site of injury in the nephritic kidneys of both the patients and the recipient monkeys, it seems likely that they are at least a contributing, if not primary, cause of the glomerular injury. The frequency of anti-GBM antibodies in human nephritis is not certain, but on the basis of preliminary observations it would appear that they are present in all cases of Goodpasture's nephritis and somewhat less than half of the cases of subacute and chronic glomerulonephritis of adults. The nature and source of immunogen stimulating the production of anti-GBM antibodies is not known, but the presence of potentially nephritogenic GBM antigens in normal urine raises the question of possible autoimmunization. From a practical point of view, it appears that patients forming anti-GBM antibodies may not be good candidates for renal transplantation since they are likely to produce in the transplants the nephritic changes already suffered by their own kidneys.
Article
Human antibodies against dextran, teichoic acid, blood group A substance, levan, tetanus toxoid, and nuclei were isolated and analyzed for their content of Gm(a), Gm(b), and Inv(a) γ-globulin genetic factors. The majority of these antibodies contained all the genetic factors determined in the donor's whole γ-globulin, but in many antibodies at very different concentrations. In a few instances specific factors could not be detected despite their presence in the individual's whole γ-globulin. Different antibodies isolated from the serum of the same individual showed different relative concentrations of genetic factors. The distribution of genetic factors seen in certain isolated human antibodies appeared to approach the selective occurrence of these factors in myeloma proteins.
Article
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 gamma-globulin in glomerular eluates compared with that in the corresponding serums. The gamma-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.
Article
This chapter discusses the chemistry and reaction mechanisms of complement. Complement constitutes the principal, immunologically relevant effector system that is present in blood serum. It consists of nine components or eleven distinct serum proteins. Membranes are the primary target of complement. They are irreversibly damaged, sustaining distinct ultrastructural lesions, by direct attack that requires participation of all nine complement components. Isolation of many of these components makes possible the analysis of the chemistry and dynamics of the complement reactions themselves and the understanding of the protein-protein interactions and enzyme activations involved. The effects of complement, primarily on cell surface membranes, eventuate in a spectrum of changes ranging from cell lysis to directed migration, histamine release, and susceptibility to phagocytosis, all of which are partially described in molecular terms. Complement research has become an active, rapidly moving, and exciting field of the biological sciences. New methods and tools are available to the biologist and clinician for the investigation of the physiogenic and pathogenic role of complement.
Article
Described here is a patient in whom the nephrotic syndrome developed during the course of secondary syphilis. A renal biopsy was performed, and the specimen showed perivascular mononuclear cell infiltration, interstitial edema and glomerular mesangial cell proliferation under light microscopy. Subepithelial hump-like electron-dense deposits with epithelial cell foot process fusion in the glomeruli were found on electron microscopy. Immunofluorescent microscopy demonstrated a granular deposition of gamma G2 lambda globulin in the glomerulus but was unable to detect complement deposition in the same location. Although the antigen involved in the glomerular deposit was not identified, the findings suggest that acute syphilitic nephrosis is an immune deposit disease.
Article
The reaction between staphylococcal protein A and human γG globulin, which is known to be mediated by structures on the Fc part of the heavy chains, was studied qualitatively using 68 isolated γG myeloma globulins. Twenty-one myeloma globulins of subgroups γG-1, γG-2 and γG-4 gave a precipitin reaction with protein A. The 35 remaining meyloma globulins of these subgroups all inhibited this reaction, indicating the presence of protein A-reactive sites. Twelve γG-3 myeloma globulins did not precipitate or inhibit the precipitation; two showed slight inhibition that could be attributed to background γG. It thus appears that anti-protein A reactivity is confined to γG-1, γG-2 and γG-4 molecules.
Article
Sera of patients with systemic lupus erythematosus were demonstrated to contain precipitating antibodies to soluble tissue components other than DNA. One dominant reaction was observed which was provisionally termed the Sm system. The antigen involved was identified in nuclei of a wide variety of cells from different species. It was associated with protein fractions but was a non-histone substance quite sensitive to periodate treatment. Antibodies to the Sm antigen could also be detected by complement fixation. They showed a high incidence in systemic lupus erythematosus with considerable specificity for the disease.
Article
Daily injections of any one of several foreign serum proteins produced in rabbits functional and morphological alterations similar to those seen in acute, subacute, and chronic human glomerulonephritis. The critical factor determining whether a rabbit would develop renal disease and the type of disease developed was the amount of antibody the rabbit formed. Those responding with much antibody were likely to develop an acute, self-limited glomerulonephritis and to be subsequently immune to further renal damage. Those responding with antibody barely sufficient to neutralize the antigen injected developed subacute and chronic glomerulonephritis. In the circulation of the rabbits with chronic glomerulonephritis, there was a daily recurring antigen-antibody reaction in the region of near antigen excess to near antibody excess which presumably led to the disease. Antigen apparently in the form of antigen-antibody complexes was deposited along the renal capillary basement membranes coincident with the development of subacute and chronic glomerulonephritis. Once developed, the morphologic stigmata of chronic glomerulonephritis persisted even after injections of antigen were stopped. However, in milder instances the renal function recovered in part after stopping antigen. This experimental model has several implications: first, the renal injury is precipitated by antigens with no known affinity for, or immunologic relationship to, kidney; second, antigen antibody complexes localize in the kidney, apparently on the basis of non-immunologic factors, and may be an etiologic agent of renal injury; third, severe hypersensitivity disorders can be related specifically to relatively poor as well as to good antibody responses; and finally, the pathogenesis suggested here offers an alternative to that of nephrotoxic serum nephritis for the experimental approach to the study of human glomerulonephritis.
Article
The primary phase of nephrotoxic serum nephritis produced by rabbit nephrotoxic serum appears to be dependent to a great extent, but not completely, upon the participation of serum complement. On the other hand, duck nephrotoxic serum produces its primary renal injury without detectable utilization of or dependence upon serum complement. The secondary phase of nephrotoxic serum nephritis appears to be largely or entirely dependent upon the host's antibody response to the heterologous gamma globulin fixed in the glomeruli. No evidence could be obtained for the existence of an autoimmune antikidney response by the host in this experimental model.
Article
Rabbit anticryoprotein and anticomplement antisera recognized a heat-labile antigen in normal human serum. This antigen best fitted the previously described US protein because of its presence in fresh human serum, euglobulin, and purified 11S preparations and its absence in heated serum, R11S, and pseudoglobulin preparations. The 11S hemolytic activity correlated well with the presence of this heat-labile antigen in the 11S region in sucrose density gradient ultracentrifugation and in the gamma globulin region on zone electrophoresis. It could be identified as a single component in the gamma globulin region in immunoelectrophoresis. The intermediate complex EAC'11S was lysed by R11S reagents and agglutinated by these antisera. The antisera also agglutinated a human complement-binding Rh-positive cell system if the 11S protein had been previously bound.
Article
Through the use of a variety of antisera to isolated myeloma proteins, four subgroups of 7S γ-globulin type proteins were readily distinguished. The first, the Vi subgroup, consisted of ten of 64 myeloma proteins studied. The second, the We group, contained the majority of myeloma proteins. The third, the Ge subgroup, included three of 50 myeloma proteins. The fourth remains ill-defined and appears heterogeneous. Counterparts for both the Vi and the Ge subgroup, were found in the Fr II γ-globulin and in the normal γ-globulin of all of a large number of individual sera studied. The unique antigenic character of both groups was localized to the H chains, although different determinants were involved for different antisera. An essential role of intact disulfide bonds was apparent with certain rabbit antisera. In addition to the special antigenic characteristics, the Ge subgroup showed in each instance a fast mobility for the F fragments produced by papain which was not found for other myeloma proteins.
Article
The antibody in agar plate test has proved valuable for the quantitative measurement of individual serum immunoglobulins. With this technique, specific antiserum is mixed uniformly in an agar gel plate. Antigen containing solutions are placed in small antigen wells cut in the agar. A concentric ring of antigen: antibody precipitate forms around the antigen well. By graphically comparing the ring diameters with those of appropriate standards, the protein concentration of the test sera can be determined. The probable error in measurement of normal serum immunoglobulins is ±10%. This procedure has been used to quantify protein concentrations as low as 0.003 mg/ml. Multiple samples can be easily tested and less than 0.1 ml of sample is required. The mean immunoglobulin levels in 20 normal human sera were found to be 12.4 mg/ml for IgG (7 S γ2-globulins); 2.8 mg/ml for IgA (γ1A or β2A-globulins) and 1.23 mg/ml for the IgM (18 S γ1M-globulins). Type K (I) and Type L (II) immunoglobulins were similarly determined. The results by the antibody-agar plate method are similar to those obtained by the isotopic immune inhibition technique, except for the serum IgA and Type K (I) and Type L (II) immunoglobulin levels. With each of these proteins the values obtained by the isotopic immune inhibition test are higher than those found by the antibody-agar plate test. The basis for this difference is discussed.
Article
Rats and guinea pigs were depleted of complement (C') by treatment with heat aggregated human γ-globulin (agg HGG), zymosan, anti-ß1C globulin, and carrageenan. Although antigen and antibody were bound to vascular structures, Arthus reactions were inhibited. This inhibition was characterized by the lack of C' binding to walls of vessels, the lack of polymorphonuclear (PMN's) cellular infiltrates, and the lack of significant vascular damage. When the same animals were followed for several hours thereafter, levels of serum C' began to rise, C' was bound in tissues, PMN infiltrates appeared, and immunologic vasculitis developed. Blood counts, chemotaxis of PMN's induced by lysates of PMN granules, together with studies on motility and phagocytosis by PMN's obtained from C' depleted rats, failed to establish any abnormality in these cells which would account for inhibition of Arthus reactions. The specificity of C' depletion in terms of effects in the first four reacting components of guinea pig C' was studied. Treatment with agg HGG led to loss of activity in all components, whereas zymosan and anti-ß1C globulin predominately affected the third component (C'3c). Carrageenan mainly affected the first two reacting components of C'. Thus, the availability of the 3c component, or a subsequently reacting component, correlated with the attraction of PMN's to immune reactants in vivo. Various antibodies with different C' fixing capacities in vitro were tested for their ability to induce immunologic vasculitis in normal animals. In rats, only those antibodies which fixed C' in vitro possessed biological activity, whereas in guinea pigs, all antibodies tested, regardless of C' fixation in vitro, induced Arthus reactions. For a given antibody in rats the vasculitis-inducing property was reflected in its ability to bind C' in vascular structures. Rats depleted of circulating PMN's by specific antibody were tested for Arthus activity. Although concentrations of immune reactants and C' were readily detected in vascular structures, no PMN infiltration occurred and significant vascular damage was averted.
Article
In acute nephrotoxic nephritis, polymorphonuclear leukocytes (polymorphs) accumulated in large numbers in the glomeruli in the first 12 hours. The endothelial cells were dislodged by the polymorphs which then came to lie immediately adjacent to the glomerular basement membranes. Ultrastructural changes in neither polymorphs nor basement membranes were observed. Depletion of polymorphs in both rats and rabbits prevented the development of proteinuria. This occurred when doses of nephrotoxic globulin were employed that produced proteinurias of as much as 1800 mg/kg/24 hours in intact rabbits, or enough to yield near maximal immediate proteinuria in intact rats. In addition, measurable glomerular damage was frequently averted until the onset of the secondary stage of NTN. Controls indicated that the polymorph depleted animals exhibited minimal non-specific changes in the blood, that the ability of their vascular beds to react to stimuli was not affected, and that deposition of nephrotoxic antibody and C' in the glomeruli was not inhibited. Elimination of polymorphs from the circulation was only partially effective in preventing glomerular damage when large doses of nephrotoxic globulin were used. This indicated that under these circumstances, a polymorph independent glomerular injury may also take place in first stage nephrotoxic nephritis. An indirect role of C', i.e., the accumulation of polymorphs, in bringing about glomerular injury in first stage nephrotoxic nephritis was apparent. When rabbit nephrotoxic globulin was injected into rats depleted of C', or when duck nephrotoxic globulin that fixed C' poorly was injected into normal rats, C' failed to bind with the antibody along glomerular basement membranes and polymorphs did not accumulate.
Article
1. A procedure was described for the electrophoretic separation and isolation of materials employing various types of supporting media. 2. Starch proved particularly useful because of its low adsorption of proteins and peptides in aqueous buffers. 3. A comparison was made of the electroosmotic flow in various media under similar conditions. 4. The separation of 1-4 cc of serum with isolation of the components could be carried out employing the starch system. 5. The α and β-lipoproteins of serum could be determined quantitatively by phospholipid and cholesterol analyses of the serum fractions.
Relation between metabolic properties and serum concentration of IgG-subclasses in man
  • A Morell
  • W Terry
  • T Waldmann
Morell, A., W. Terry, and T. Waldmann. 1969. Relation between metabolic properties and serum concentration of IgG-subclasses in man. Clin. Res. 17: 356. (Abstr.)
Fuden-berg Biologic globulin. VIII. Aggregated ferent classes
  • T Ishizaka
  • K Ishizaka
  • S Salmon
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