Anti-gamma Globulin Research Articles

Anti-immunoglobulins and their idiotypes: are they part of the immune network?

The anti-gamma globulins represent a very heterogeneous group of proteins with widely different specificities that might be considered a portion of the immune network. The hypothesis is presented that at least some of these proteins have other specificities, with the anti-gamma globulin reactivity being a secondary property. Anti-idiotypic antibodies are possible candidates. This is based on the low binding affinity for gamma globulin of many of these proteins and the results of CRI and sequence studies. All the proteins of the major CRI group have VKIIIb light chains, and these have a dominant but not total influence on this CRI. This has been evident from studies with rabbit antibodies and recently also with monoclonal hybridoma antibodies. Sequence studies have also demonstrated the similarity in light chains that, however, are not greater than with other VKIIIb chains; the heavy chains show little similarity except possibly in the J segment. The possibility is discussed that antibodies with secondary anti-gamma globulin binding properties might have selective advantages.

Immunosuppressive effect of Entamoeba histolytica extract on hamsters.

The immune response to sheep red blood cells (SRBC) in mice and hamsters injected with an extract of entamoeba histolytica was studied. Both the primary and secondary immune response, measured by anti-SRBC antibody titers, were unaltered in the mouse, while a significant depression of the primary, but not the secondary, response was observed in the hamster. The effect was greatest when the amebic extract (AE) and SRBC were injected on the same day. The number of anti-SRBC rosettes formed in the spleen cells of hamsters treated with both AE and SRBC on day 0 was measured from days 1-16. The response peaked on day 13, while cells from animals injected with SRBC alone gave a maximal response on day 5. The formation of anti-SRBC rosettes in T-lymphocyte-enriched spleen cells treated with anti-gamma globulin serum and complement was almost abolished for the duration of the experiment. It is suggested that the mechanism responsible for this immunosuppressive phenomenon could involve early interference in the afferent limb of the immune response.

Anti-immunoglobulin antibodies. III. Properties of sequential anti-idiotypic antibodies to heterologous anti-gamma globulins. Detection of reactivity of anti-idiotype antibodies with epitopes of Fc fragments (homobodies) and with epitopes and idiotopes (epibodies).

Murine anti-V region antibodies against a human monoclonal protein Gl with anti-gamma-globulin activity and bearing the Wa cross-reactive idiotype were prepared in several strains of mice. Antibodies were obtained that were specific for the Gl idiotype, the Wa cross-reactive idiotype, and for various framework antigenic determinants that were distinguished by a variety of procedures. Synthesis of such antibodies were found to be independent of MHC and Igh gene complexes. These anti-V region antibodies, produced by a majority of mouse strains investigated, also share a cross-reactive idiotype recognized by BALB/c anti-anti-V region Gl antibodies. A fraction of BALB/c anti-anti-V region Gl antibodies displayed human Fc gamma binding activity and, therefore, can be considered homobodies, as described in other systems. Two of the anti-idiotype antibodies obtained in this system exhibited a peculiar property: they interacted not only with their own antigen (IgM Gl), but also with the Fc fragment with which IgM Gl reacts. Thus, A/J anti-V region Gl antibodies bind to the human Fc gamma fragment in addition to IgM Gl. Similarly, a monoclonal CB6 F1 anti-anti-V region Gl antibody interacted with the V region of IgM Gl as well as with the syngeneic anti-V region antibodies. We called these anti-idiotypic antibodies epibodies because they interact with epitopes of the primary antigen. These homobodies and epibodies, obtained by immunization across heterologous barriers, represent new examples of recognition of the internal image of the antigen within the immune system.

Complete amino acid sequence of variable domains from two monoclonal human anti-gamma globulins of the Wa cross-idiotypic group: suggestion that the J segments are involved in the structural correlate of the idiotype.

The complete amino acid sequences of the variable regions of two human IgM anti-gamma globulins are reported. The two proteins, Sie and Wol, share idiotypic determinants that are distinct from those shared by the two previously sequenced IgM anti-gamma globulins, Lay and Pom [Capra, J. D. & Kehoe, J. M. (1974) Proc. Natl. Acad. Sci. USA 71, 4032-4036; Klapper, D. G. & Capra, J. D. (1976) Ann. Immunol. (Inst. Pasteur) 127C, 261-271.] In contrast to Lay and Pom, Sie and Wol have very little homology in the complementarity-determining hypervariable regions of either their light or heavy polypeptide chains. However, there are only two amino acid differences in the framework portions of the light chains, and the J segments of both chains are homologous. These data suggest that the predominant idiotypic determinant in this cross-idiotypic system may be different than in the Po group and may be either related to their light chain framework structures or to the amino acid sequence in the J segments of these molecules.

Enzyme linked immunoassay and fluorescent antibody techniques in the diagnosis of viral diseases using staphylococcal protein—A instead of anti-gamma-globulins

Staphylococcal protein—A (SpA) is known to interact with the crystallizable fragment (Fc) of IgG molecules from several species. In the present study, SpA coupled to either fluorescein isothiocyanate (FITC) or peroxidase was used in place of antisera to IgG for the fluorescent antibody (FA) techniques and the enzyme linked immunoassay (ELISA). The SpA conjugates produced low background staining when applied in these techniques, and provide a rapid, highly specific and sensitive means for the identification of viral isolates and the detection of serum antibodies. Moreover, SpA is a single reagent that replaces various preparations of anti-gamma globulin against many species. SpA-FITC conjugate was successfully applied for the identification of pseudorabies virus, hog cholera virus, swine vesicular disease virus, transmissible gastroenteritis virus, porcine parvovirus and porcine enteroviruses. Antibody titers against the mentioned viruses could be determined semi-quantitatively in the indirect FA test with SpA-FITC. In our laboratory the ELISA became a routinely practicable serological test for the detection of antibodies only after we introduced SpA-peroxidase as a marker for the IgG.

Rheumatoid factor in melanoma patients:Alterations of humoral tumor immunity in vitro

Rheumatoid factors (RF) were associated with alterations of antibody reactions to melanoma cells in vitro by two serologic assays. Removal of RF from melanoma patients' sera by absorption with Cohn's Fraction II coated latex particles enhanced seroreactivity in the Immune Adherence (IA) assay and diminished IgM detection by the Indirect Membrane Immunofluorescence (IMI) assay. The addition of serum with high titers of RF to these assay systems led to diminution of IA reactivity and enhancement of IgM detection by IMI. Since these factors are found in cancer patients' sera and can alter humoral immune reactions directed against antigens on the membranes of tumor cells, their presence should be recognized when performing assays with tumor target cells. RF may be of significance in the host-tumor relationship in vivo.

Antigamma globulin factors and levels of IgG, IgA and IgM in sera of patients with Dupuytrenʼs contracture

Antigamma globulin factors and levels of IgG, IgA and IgM in sera of patients with Dupuytrenʼs contracture

Insulin kinetics in metasomatotropic diabetes

Insulin kinetics were studied in dogs with metasomatotropic diabetes that had been induced by administration of growth hormone. The duration of this severe diabetes was about 1 yr. One of the dogs had been under insulin treatment for about 1 mo, but three others had not received insulin. In the unanesthetized diabetic and control dogs, in the postabsorptive state following intravenous injection of 131I-insulin, the amount of labeled immunoreactive insulin (L-IRI) injected and the amount present in venous blood were determined as the radioactivity of the complex precipitated by addition of excess of anti-insulin serum and anti-gamma globulin serum. The metabolic clearance rate of L-IRI was not significantly different in the diabetic and normal dogs (13.1 ± 3.3 and 10.7 ± 1.6 ml/kg body weight/min, respectively); the difference in fractional rate of disappearance was also not significant (0.053 and 0.054 fraction/min, respectively). The volume of distribution of L-IRI was comparable to the expected volume of extracellular fluid. The mass rate of disappearance of unlabeled immunoreactive insulin (IRI) in the metasomatotropic diabetic dogs was about one-half normal, however, due to the lower serum IRI level. The diabetic and control dogs were in a steady state with respect to systemic serum insulin during the test; thus the rate of loss could be related to the basal systemic delivery of insulin. The sub-normal delivery of insulin was sufficient for maintenance of life over long periods in this diabetic state, although the physical condition of the diabetic dogs deteriorated slowly.

IDENTIFICATION OF PARAMYXOVIRUS‐SPECIFIC HAEMOLYSIS‐INHIBITING ANTIBODIES SEPARATE FROM HAEMAGGLUTINATING‐INHIBITING AND NEURAMINIDASE‐INHIBITING ANTIBODIES

Egg-grown Newcastle disease (NDV) and mumps virus were used for preparation of rabbit hyperimmune sera against purified whole virus and projectionless virus particles. These sera and convalescent sera after natural NDV and mumps infections in chickens and human subjects, respectively, were studied in haemolysis-inhibition (HLI), haemagglutination-inhibition (HI) and neuraminidase-inhibition (NI) tests both before and after absorption with Tween 80-ether (TE) treated virus preparations. In addition, neutralization tests using the different sera were carried out. HI and NI antibodies and the major population of neutralizing antibodies in convalescent sera were removed by absorption with TE treated virus material without changing the titre of non-HI HLI antibodies. Rabbit hyperimmune sera directed against projectionless virus particles exhibited HLI antibody titres in marked excess of HI and NI antibody titres, whereas this was not found in sera against whole virus. Absorption with TE treated virus material resulted in removal of all demonstrable antibody activities in sera against whole virus. The corresponding absorption of sera against projectionless particles eliminated HI antibodies without changing the titre of non-Hi HLI antibodies. In rabbit hyperimmune sera, HI antibodies were of primary importance in neutralization tests. After addition of anti-gamma globulin to the test, an efficient neutralization was observed if mumps non-HI HLI antibodies were used whereas this was not found if NDV non-HI HLI antibodies were used.

Cryoimmunoglobulinemia in rheumatoid arthritis. Significance in serum of patients with rheumatoid vasculitis.

Cryogloculins were examined in a standardized manner in an unselected group of 35 patients with rheumatoid arthritis (RA) and 8 patients with RA complicated by cutaneous vasuclitis and neuropathy. Optimum conditions for detection and characterization of cryoglobulins were established; the proportion of resolubilized to total precipitable protein remained constant in an individual patient under these conditions. All 8 vascultis patients and 9 of 35 other patients with RA exhibited cryoglobulins; total protein and immunoglobin content were significantly higher in the cryoglobulins of patients with vasculitis. Immunoglobulins G and M constituted two-thirds and three-quarters of the total protein in the cryoglobulins from uncomplicated rheumatoid and vasculitis patients, respectively. Serum antiglobulin titers were higher, and serum C3 levels were lower, in vasculitis patients compared to rheumatoid patients without vasclitis. Anti-gamma globulin activity was detected in all cryoglobulins from vasculitis patients. Cryoglobulin IgG and IgM were polyclonal. Density gradient analyses demonstrated the majority of the cryoglobulin activity to reside in the 19S IgM fraction. There was no evidence of a light weight (8S) IgM. A monoclonal rheumatoid factor did not detect 7S-ANTI-7S complexes in the cryoprecipitates, but acid eluates from some cryoglobulins absorbed with insoluble IgG revealed an antiglobulin of the IgG class. Serial studies performed on vasculitis patients treated with cyclophosphamide disclosed a relationship between clinical evidence of vasculitis and the presence of cryoglobulins. The antigen (IgG) and antibody (largely IgM rheumatoid factor) nature of these cryglobulins is presented as evidence that the widespread vascular complications of RA are mediated, at least in part, by circulating immune complexes.

Structure of antibodies with shared idiotypy: the complete sequence of the heavy chain variable regions of two immunoglobulin M anti-gamma globulins.

The complete amino acid sequence of the heavy chain variable regions of two different molecules of immunoglobulin M anti-gamma globulin has been determined. These proteins, from different human patients, had independently been shown to share idiotypic specificity. Only eight sequence differences were discernible for the entire length of their heavy chain variable regions. Five of the differences occurred outside hypervariable regions, while three were placeable within such regions. A comparison of these molecules of anti-gamma globulin with the seven human V(H)III variable region sequences presently available for immunoglobulins without known antibody activity showed that the great majority of sequence differences between the two idiotypically similar antibodies and these seven proteins were confined to hypervariable regions. This study illustrates in precise terms a convergence of the distinct immunological properties of idiotypy, hypervariable region structure, and combining site specificity as they relate to the variable region of the immunoglobulin molecule. To a great degree these properties now appear to be a reflection of the same structural attributes of the variable region.

Immunocytochemical localization of LHRH in axons and nerve terminals of the rat median eminence.

Cytoimmunological localization of LHRH-containing axons and terminals in the median eminence of male rats was achieved with a potent anti-LHRH antiserum and subsequent labeling with a second, peroxidase- or fluorescein isothiocyanate-bound, anti-gamma globulin antibody. The distribution of peptidergic fibers extends from the rostral part of the median eminence, immediately behind the retrochiasmatic area, to the region of the pituitary stalk. In the external zone of the rostral median eminence and close to the stalk, the antigenic material appears in the form of large granules accumulated around blood vessels. This distribution suggests that such granules are likely to represent axon terminals, in contrast to axonal processes containing smaller granular elements lined up radially or longitudinally underneath the external zone.

STAPHYLOCOCCAL RADIOIMMUNOASSAY FOR HEPATITIS B ANTIGEN AND ANTIBODY

A rapid radioimmunoassay (RIA) for the measurement of hepatitis B antigen and antibody is described in which protein A carrying Staphylococcus aureus is used as a solid phase antigamma globulin reagent. The tests are easy to perform, they are completed in two hours, and are easily automatized. The competition principle is used for antigen testing while antibody is measured by direct binding of radiolabelled hepatitis B antigen. Using human reconvalescent antisera, the staphylococcal test for antigen in 25 μl serum has about the same sensitivity as the Ausria test. The staphylococcal test for antibody has at least the sensitivity of the passive hemagglutination test. Both tests are highly specific.

Binding of antinucleoside antibody to DNA

Reactions between radiolabeled heat-denatured E. coli DNA and antibodies specific for each of the four naturally occurring mononucleosides were detected by the modification of the Farr procedure using antigamma globulin to precipitate the antigen-antibody complex. Nonimmune serum binds less than 5 per cent of the radiolabeled DNA. The homologous hapten is at least 10 3 times more inhibitory than the heterologous haptens. Formaldehyde-denatured DNAs are also effective inhibitors. M. lysodeikticus DNA‡ (72 per cent G-C), denatured in the presence of 1% formaldehyde inhibited anti-cytidine better than did Cl. perfringens DNA (30 per cent G-C) similarly denatured. The reverse was obtained with antithymidine. Thus DNAs of varying base composition can be differentiated from each other.

Immunofluorescent study of placental bed vessels in pre-eclampsia of pregnancy

Anti-gamma globulin and anticomplement fluorescent stains of decidual vessel lesions were positive in women with pre-eclampsia. The lesions may represent an immunopathologic process which could lead to diminished perfusion of the placenta.

Anti-gamma globulins and chronic infection: comparative studies of the immune response to various bacteria and gamma globulin preparations.

A study of the relationship of clinical states associated with prolonged infection (bacterial endocarditis and osteomyelitis) and generation of serum anti-gamma globulins was made with particular reference to quantitative amounts of staphylococcal protein A in various infecting strains. No correlation between individual strain amounts of protein A and presence of anti-gamma globulins was detected. Thirty-eight rabbits were immunized intravenously with various strains of bacteria (Staphylococcus aureus, enterococci, Streptomyces viridans, pneumococci, pseudomonas, and Escherichia coli) for periods of 6 weeks, and antibacterial as well as anti-gamma globulin antibodies were assayed. No single group or strain of bacteria stood out as being more prone to produce anti-gamma globulins than others tested. Most rabbits developed anti-gamma globulins reacting with human gamma globulins, whereas the specificity for rabbit gamma globulin appeared more restricted. In 16 rabbits immunized with eight different strains of S. aureus, quantitative elevation of serum gamma globulin above 2.5 g per 100 ml often seemed to be correlated with presence of detectable serum anti-gamma globulins. By contrast 15 rabbits immunized with autologous or isologous rabbit gamma globulins in many instances developed extremely high titers of anti-gamma globulins showing primary specificity for human rather than rabbit gamma globulin. These studies further amplify the remarkably heterogeneous anti-gamma globulin reactivity associated with various types of immune response.

Appearance and persistence of antibodies against different virus components after regular measles infections.

Different measles virus-specific antibody activities in acute, early (11 to 40 days after rash) and late (4 to 20 years postinfection) convalescent sera and gamma globulin were determined. Early immunoglobulin G antibodies gave a poor neutralization, which was increased 10- to 60-fold by addition of anti-gamma globulin. There was a high degree of correlation between titers of hemolysis-inhibiting (HLI) and hemagglutinating-inhibiting (HI) antibodies. However, in one out of fifteen late convalescent sera an HLI antibody titer of 640 in the presence of titer of only 20 in HI tests with Tween 80-either-treated antigen was found. Similar findings were made with sera from two patients with multiple sclerosis included in a parallel study. A somewhat higher titer of HI antibodies was demonstrable in these three sera when untreated material was used as antigen. These findings are interpreted in the following way. Antibodies against the hemagglutinin can block not only virus-specific agglutination but also lysis of red cells. In contrast, antibodies against the hemolysin, besides blocking the biological activity of this component, carry only a slight HI activity. This HI activity can be detected only by use of antigen preparations containing hemagglutinin-associated hemolysin.Complement-fixation (CF) and immunodiffusion tests (the latter were carried out with antigen preparations treated with 0.25% sodium dodecyl sulfate) demonstrated that, in almost all cases, antibodies against nucleocapsid structures dominated quantitatively among antibodies appearing in connection with and persisting after regular measles infections. Generally, only low titers of antibodies reacting with purified small particle hemagglutinin (HA; 10 to 14S) or additional structural or nonstructural components were identified in CF and immunodiffusion tests.

Equine Nfectious Anemia: Detection of Infectious Virus-Antibody Complexes in Me Serum

The sera of horses with persistent infection by equine infectious anemia virus were examined for the presence of infectious virus-antibody complexes, Virus-containing sera were treated with rabbit anti-equine gamma globulin, rabbit anti-equine albumin, and normal rabbit serum. Infectivity remaining in the supernates was titrated in primary equine leukocyte cultures. In 2 of 3 sera anti-gamma globulin precipitated at least 2 log10 of viral infectivity using the anti-albumin and normal rabbit serum treatments as controls. This demonstrates that in some sera EIA virus circulates as an immune complex with antibody, possibly enhancing its ability to persist in the host by providing a continued source of virus for reinfection.

Structure-function relationships among anti-gamma globulin antibodies.

Annals of the New York Academy of SciencesVolume 190, Issue 1 p. 371-381 STRUCTURE-FUNCTION RELATIONSHIPS AMONG ANTI-GAMMA GLOBULIN ANTIBODIES* J. Donald Capra, J. Donald Capra Department of Microbiology Mount Sinai School of Medicine of the City University of New York and The Rockefeller University New York, N.Y.Search for more papers by this authorJ. Michael Kehoe, J. Michael Kehoe Department of Microbiology Mount Sinai School of Medicine of the City University of New York and The Rockefeller University New York, N.Y.Search for more papers by this authorRobert J. Winchester, Robert J. Winchester Department of Microbiology Mount Sinai School of Medicine of the City University of New York and The Rockefeller University New York, N.Y.Search for more papers by this authorHenry G. Kunkel, Henry G. Kunkel Department of Microbiology Mount Sinai School of Medicine of the City University of New York and The Rockefeller University New York, N.Y.Search for more papers by this author J. Donald Capra, J. Donald Capra Department of Microbiology Mount Sinai School of Medicine of the City University of New York and The Rockefeller University New York, N.Y.Search for more papers by this authorJ. Michael Kehoe, J. Michael Kehoe Department of Microbiology Mount Sinai School of Medicine of the City University of New York and The Rockefeller University New York, N.Y.Search for more papers by this authorRobert J. Winchester, Robert J. Winchester Department of Microbiology Mount Sinai School of Medicine of the City University of New York and The Rockefeller University New York, N.Y.Search for more papers by this authorHenry G. Kunkel, Henry G. Kunkel Department of Microbiology Mount Sinai School of Medicine of the City University of New York and The Rockefeller University New York, N.Y.Search for more papers by this author First published: December 1971 https://doi.org/10.1111/j.1749-6632.1971.tb13549.xCitations: 36 † Supported in part by National Science Foundation grant GB-17046 and National Institutes of Health grant AI-09810. AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature Volume190, Issue1ImmunoglobulinsDecember 1971Pages 371-381 RelatedInformation

Subacute Bacterial Endocarditis as an Immune Disease

Among the numerous immunologic changes occurring in s BE, one of the most significant appears to be the development of anti-gamma globulin (rheumatoid) factors in many patients. These factors react with 7S opsonic gamma G antibodies to inhibit phagocytosis of the invading bacteria; they may thus hold the key to the chronicity of the disease. Relevantly, with effective treatment of SBE, antiglobulin factors disappear.