Normal Γ-globulin Research Articles

ANTIGENIC UNIQUENESS OF BENCE JONES PROTEINS.

MUCH information about the structure of the human immunoglobulins has derived from investigations of the antigenic characteristics of Bence Jones proteins. The two major antigenic groups of Bence Jones proteins have immunological counterparts in the light (L) chains of the normal population of γ2-, γ1A- and γ1M-globulins1. Using group specific rabbit antisera, we have shown that Bence Jones proteins of the same immunological group from different patients contain antigenic determinants not overtly present in pooled human 7S γ-globulin from normal individuals2. It is probable that some degree of the antigenic uniqueness of Bence Jones proteins is related to the individual antigenic specificity of single immunizing proteins2. Another variable determining at least a part of the immunological difference between Bence Jones protein and normal unreduced γ-globulin is related to the unmasking of hidden antigenic determinants of L chains when they are either not incorporated into or cleaved from the parent γ-globulin molecule3. It has been suggested that certain potential antigenic sites of L-polypeptide chains are inaccessible in the intact γ-globulin molecule due to steric hindrance of adjacent H chain4.

STRUCTURAL ASPECTS OF HUMAN ERYTHROCYTE AUTOANTIBODIES. I. L CHAIN TYPES AND ELECTROPHORETIC DISPERSION.

The 7S gamma-globulins causing erythrocyte autosensitization in 20 patients were isolated by elution and examined for homogeneity or heterogeneity of their L chain types and electrophoretic dispersion. The isolated erythrocyte autoantibodies from 12 patients contained only 1 detectable L chain type. Two of these "monotypic" populations showed appreciable restriction of electrophoretic dispersion, while 2 others more nearly resembled the electrophoretic heterogeneity of normal gamma-globulins. The autoantibodies from the other 8 patients exhibited L chains of both types. The single "bitypic" population so tested was relatively polydisperse electrophoretically. As a comparison, anti-Rh(o) isoantibodies from 5 of 6 donors without known hematologic disease showed bitypic reactions, and 2 of these isoantibody populations were relatively polydisperse electrophoretically. One Rh isoantibody is described which contained only 1 demonstrable L chain type. The structural similarities to "paraproteins" observed in a significant proportion of these erythrocyte autoantibodies raise the possibility of their origin from a restricted population of antibody forming cells, and may have implications concerning the pathogenesis of erythrocyte autosensitization.

GM CHARACTERS OF 7 S GAMMA-MYELOMA PROTEINS AND CORRESPONDING INDIVIDUAL NORMAL GAMMA-GLOBULINS.

Acta Pathologica Microbiologica ScandinavicaVolume 61, Issue 2 p. 181-189 Article GM CHARACTERS OF 7 S γ MYELOMA PROTEINS AND CORRESPONDING INDIVIDUAL NORMAL γ-GLOBULINS† ULF NILSSON, ULF NILSSON From the Department of Clinical Chemistry, Malmö General Hospital, Malmö, University of Lund, SwedenDr. Nilsson's present address is: Division of Experimental Pathology, Scripps Clinic and Research Foundation, La Jolla, California.Search for more papers by this author ULF NILSSON, ULF NILSSON From the Department of Clinical Chemistry, Malmö General Hospital, Malmö, University of Lund, SwedenDr. Nilsson's present address is: Division of Experimental Pathology, Scripps Clinic and Research Foundation, La Jolla, California.Search for more papers by this author First published: September 1964 https://doi.org/10.1111/apm.1964.61.2.181Citations: 4 †Aided by grants from the Medical Faculty of Uppsala and from Österlundska Stiftelsen, Malmö. 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 onFacebookTwitterLinked InRedditWechat Citing Literature Volume61, Issue2September 1964Pages 181-189 RelatedInformation

H CHAIN SUBGROUPS OF MYELOMA PROTEINS AND NORMAL 7S γ-GLOBULIN

Through the use of a variety of antisera to isolated myeloma proteins, four subgroups of 7S gamma-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 gamma-globulin and in the normal gamma-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.

Contribution of γ-globulin subunits to electrophoretic heterogeneity: Identification of a distinctive group of 6·6 S γ-myeloma proteins

The electrophoretic heterogeneity of 6.6 S γ-globulins was investigated by studying the subunits obtained by reduction and alkylation and by papain digestion of normal γ-globulins and 15 γ-myeloma proteins. Studies of normal γ-globulin fractions and 12 γ-myeloma proteins revealed that the relative electrophoretic mobility of the H chains (reduction and alkylation) and the S pieces (papain treatment) corresponded to that of intact proteins. Evidence was obtained that the part of the H chain present in the S piece (termed the A piece by Porter) is largely responsible for the electrophoretic differences between γ-globulins. A group of 6.6 S γ-myeloma proteins were distinguished from other γ-myeloma proteins and from the bulk of normal γ-globulins on the basis of the response to papain digestion. Evidence is reviewed which indicated that these γ-myeloma proteins have distinctive H chain properties.

CONJUNCTIVAL BLOOD FLOW IN CRYOGLOBULINEMIA.

Introduction Cryoglobulins are groups of cold-precipitable proteins greatly resembling normal γ-globulin,1and their most striking characteristic is their ability to precipitate when exposed to a lowered temperature. An essential part of the phenomenon relates to the precipitate returning to solution when the temperature is raised to 37 C. The most prominent symptom noted in this disease is a particular sensitivity to lowered temperature with oronasal bleeding a common finding.2Physical findings include weight loss, pallor, petechiae with purpura, mottling and occasional ulceration of the lower extremity. The ocular findings include slowing and stasis of blood flow with marked erythrocytic aggregations in the vessels of the bulbar conjunctiva.3Ophthalmoscopic study has demonstrated dilated retinal veins with slowing and segmentation as well as occasional retinal hemorrhages and exudates.4 In 1957 Harders5reported observations on conjunctival blood flow in a patient with Waldenstrom's macroglobulinemia who in addition

THE TRANSMISSION OF ANTIBODIES AND NORMAL GAMMA-GLOBULINS ACROSS THE YOUNG MOUSE GUT.

When 11-day mice are fed with homologous or heterologous antibodies or with heterologous γ-globulin, the maximum concentration of these proteins in their sera is reached 2 h later, irrespective of the amount or specificity of the proteins administered, or of the final concentration achieved. The relation between y ( = relative titre of antibody, or concentration of heterologous γ-globulin, attained in the serum) and x ( = dose of γ-globulin administered) is best represented by ( a/y — b/x ) = 1, a rectangular hyperbola with a theoretical maximum of y max . = a when x is very large. For any x , the proportion of y max . attained depends inversely on the value of b which is greatest for rat γ-globulin and then falls progressively for mouse anti- Salmonella pullorum agglutinins, bovine γ-globulin, guinea-pig γ-globulin, rabbit anti- Brucella abortus incomplete agglutinins, rat anti- Salm . pullorum agglutinins, rabbit γ-globulin and guinea-pig anti- Salm . pullorum agglutinins. The lower the value of b for any protein the less is the transmission of that protein affected by admixture with other γ -globulins, and if the protein is a γ-globulin, the more it reduces the transmission of antibodies or of other γ-globulins mixed with it. The value of b for rat anti- Salm . pullorum agglutinins is less than that for the corresponding normal γ-globulin. Fragment III of rabbit γ-globulin, prepared by the digestion method of Porter, is transmitted across the gut less readily than the intact molecule, but its ability to reduce the transmission of antibody is greater. These observations are consistent with the hypothesis of competitive absorption of ingested γ-globulin by a specific receptor within the absorptive cells of the gut. The receptor protects the attached protein from proteolysis and conveys it to the blood or lymph, possibly altered to a form more acceptable to the host.

A Comparison of Fragments of Rabbit Antibodies and Normal γ-Globulin by the Peptide-Map Technique*

A Comparison of Fragments of Rabbit Antibodies and Normal γ-Globulin by the Peptide-Map Technique^*

Occurrence of Myeloma-Like γ-Globulin in C.S.F of A Four-Month Old Infant with Hydrocephalus

Myeloma-like immune globulins present themselves as narrow bands upon paper electrophoresis, and usually show a characteristic appearance in immunoelectrophoresis. Two antigenically different groups of myeloma proteins have been described: groups I and II. Recently, 60% of normal γ-globulin, throughout the mobility range of γ-globulin, has been shown to possess the antigen characteristic for group I, and 30% that for group II myeloma. Occurrence of myeloma-like proteins in the serum is not restricted to multiple myeloma. They may also be seen with other tumors, such as reticulum cell sarcoma, and various carcinomas. In addition, Sonnet and Milhaux have reported on the frequent occurrence of myeloma-like ("monoclonal") γ-globulins in the serum of adult Bantus with different diseases. When large amounts of a myeloma protein are present in the serum, it may be found in a much lower concentration in the spinal fluid.

Two Types of 6.6 S γ-Globulins, β2A-Globulins and 18 S γ1-Macroglobulins in Normal Serum and γ-Microglobulins in Normal Urine

Summary Two types of γ-globulin antigenic determinants (I and II) are described. These determinants are common to all γ-globulin groups. The 6.6 S γ-globulins, γ1-macroglobulins, and β2A-globulins of normal serum, and the γ-microglobulins of normal urine were divided into two types by the presence of type I or II antigenic determinants. Individual γ-globulin molecules appear to have either type I or II, but not both, types of antigenic determinants. All individual sera tested had both type I and II γ-globulin molecules. In normal serum the type I γ-globulins predominate in a ratio of about 2:1. Similar ratios were observed in normal serum 6.6 S γ-globulins and γ1-macroglobulins, but more nearly equal ratios of type I and II molecules were found in the γ-microglobulins of normal urine. Type I and II γ-globulin molecules are similarly distributed throghout the entire range of electrophoretic mobility of normal γ-globulin. Type I and II γ-globulin molecules also are similar in ultracentrifugal and chromatographic behavior and in hexose content. Preliminary observations, however, indicate that the InV genetic factors are more likely to be on type I than on type II molecules.

TWO MAJOR TYPES OF NORMAL 7S γ-GLOBULIN

Normal 7S human gamma-globulin was found to contain two fundamental antigenic groups of molecules. The group 1 molecules of normal gammaglobulin correspond antigenically to group 1 multiple myeloma proteins and Bence Jones proteins; and group 2 molecules of normal gamma-globulin correspond antigenically to group 2 multiple myeloma proteins and Bence-Jones proteins. Among pooled human Fr II and several individual gamma-globulin preparations, approximately 60 per cent of molecules belong to group 1 and approximately 30 per cent of molecules to group 2 in this classification. The possible existence of a third minor antigenic group, constituting about 10 per cent, is discussed. Antisera to Bence Jones proteins of antigenic group 1 and group 2, in conjunction with I-131-labeled 7S gamma-globulin proved to be the most useful system for defining the antigenic groups of normal gamma-globulin. The group-specific antigenic determinants of normal 7S gamma-globulin molecules were located on the S fragments of these proteins.

Structural Transitions in Antibody and Normal γ-Globulins: III. IODINATION OF RABBIT γ-GLOBULIN

Structural Transitions in Antibody and Normal γ-Globulins: III. IODINATION OF RABBIT γ-GLOBULIN

ANTIGENIC RELATIONSHIPS OF BENCE JONES PROTEINS, MYELOMA GLOBULINS, AND NORMAL HUMAN γ-GLOBULIN

By means of immunodiffusion and immunoelectrophoresis study has been made of antigenic relationships of Bence Jones proteins, and the three classes of normal and pathological immunoglobulins, 7S gamma, beta(2A), and beta(2M). All thirty-nine Bence Jones proteins studied could be classified into either one of two distinct antigenic types, A or B. Both types are related to the immunoelectrophoretically slow (S) fragment of a papain digest of normal gamma-globulin; B is related more closely than A, but neither has antigenic determinants in common with the fast (F) fragment. The 7S gamma myeloma globulins were either immunological type I or II. The papain digests of these proteins produced the S and F precipitin lines in immunoelectrophoresis but multiple bands in starch gel electrophoresis, especially in the F region. The S fraction of type I myeloma globulins is antigenically similar to Bence Jones protein of type B, and the S component of type II myeloma globulins has antigenic determinants in common with type A Bence Jones protein. Correspondingly, myeloma patients with type I globulins and proteinuria usually excrete type B Bence Jones proteins, whereas patients with type II excrete type A proteins. The F fragment is the part common to normal 7S gamma-globulin and types I and II myeloma globulins but is absent in beta(2A) and beta(2M) pathological globulins and in both types of Bence Jones proteins. Papain digests of beta(2A) myeloma globulins produced a single precipitin line in immunoelectrophoresis. beta(2A) myeloma globulins appeared to have two antigenic units, one in common with type B Bence Jones protein and normal gamma-globulin, and another specific to beta(2A). The beta(2A) myeloma patients excreted type B Bence Jones protein. The papain digest of a macroglobulin produced two precipitin lines, the faster of which had antigenic determinants in common with type B Bence Jones protein, the slower seemed specific for the macroglobulin. Five serum micromolecular globulins proved to be either type A or B Bence Jones proteins. From the above results, an antigenic map was constructed showing which determinants are shared and which are specific for normal 7S gamma-globulin, types I and II myeloma globulins, beta(2A) myeloma globulins, a macroglobulin, and types A and B Bence Jones proteins.

Genetic characters of human gamma-globulins in myeloma proteins.

The genetic factors Gm(a), Gm(b), Gm(x), and Inv(a), Inv(b) described for normal human gamma-globulin were all found in different myeloma proteins. A single myeloma protein never contained more than one product of alternate alleles even in heterozygous individuals. However, factors determined by the two different loci were often found in the same myeloma protein. The Gm(a) character of the myeloma protein parallelled that of the normal gamma-globulin of the same serum in most cases. In contrast, the Gm(b) character was usually absent in the myeloma protein when it was directly demonstrable in the normal gamma-globulin. The myeloma proteins from six Negroes were Gm(a+b-), whereas the normal gamma-globulin was Gm(a+b+). This indicates that the effect of gene Gm(b) is similar in Negroes and whites, even though its relation to gene Gm(a) is different in the two races. Gm factors were found only in the 7S gamma-globulin type myelomas and not in other products of plasma cell tumors. Inv characters were, however, present in all four types of proteins studied, namely 7S and 19S gamma-globulins, beta(2A)-globulins, and Bence Jones proteins. In two instances, genetic heterogeneity of the protein products was demonstrated suggesting the proliferation of more than one clone of plasma cells in some multiple myeloma patients. The accumulated evidence obtained in this study strongly suggested that the presence and absence of genetic characters was compatible with the concept that myeloma proteins were closely analogous to individual moieties in the spectrum of normal gamma-globulins rather than truly abnormal proteins. Their study offered evidence of a heterogeneity of genetic characters among the normal gamma-globulins in a given individual. It also appears probable that in normal individuals single plasma cells have a restricted capacity to express genetic information in their protein product.

Chemical differences between antibody fragments as shown by paired label studies.

PORTER1has shown that antibodies can be digested with papain to yield univalent fragments, molecular weight about 50,000, and an inactive fragment, molecular weight 80,000. The univalent fragments obtained are separable by chromatography on carboxymethylcellulose (CMC) into two fractions called Fraction I and Fraction II. Since then, Stelos et al.2have shown that the fragments in each fraction come from different antibody molecules (see also Palmer et al.3). We have also been able to show that fragments of Fraction I and fragments of Fraction II can be further fractionated and that the fragments in the various fractions appear to be different on a chemical basis also2. We showed that in the case of specifically purified antibody against the azobenzoate ion and azobenzenearsonate ion the rates of iodination of the fragments appearing in Fraction I are different from those appearing in Fraction II. This was demonstrated by iodinating the mixture of fragments and then chromatographing it. It was found that the iodine–protein ratio was different for the two fractions. On the other hand, the rates of iodination of the fragments of Fraction I and of Fraction II from normal γ-globulin were usually quite similar. The results of this experiment indicate that not only were there chemical differences between the fragments in Fraction I and in Fraction II from a specifically purified antibody, but also that these molecules differed from normal γ-globulin.

Structural Transitions in Antibody and Normal γ-Globulins. I. Molecular Properties

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTStructural Transitions in Antibody and Normal γ-Globulins. I. Molecular PropertiesHarold. Edelhoch, R. E. Lippoldt, and R. F. SteinerCite this: J. Am. Chem. Soc. 1962, 84, 11, 2133–2138Publication Date (Print):June 1, 1962Publication History Published online1 May 2002Published inissue 1 June 1962https://doi.org/10.1021/ja00870a027RIGHTS & PERMISSIONSArticle Views24Altmetric-Citations22LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit PDF (691 KB) Get e-Alerts Get e-Alerts

Structural Transitions in Antibody and Normal γ-Globulins. II. Fluorescence Polarization Studies

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTStructural Transitions in Antibody and Normal γ-Globulins. II. Fluorescence Polarization StudiesR. F. Steiner and Harold. EdelhochCite this: J. Am. Chem. Soc. 1962, 84, 11, 2139–2148Publication Date (Print):June 1, 1962Publication History Published online1 May 2002Published inissue 1 June 1962https://doi.org/10.1021/ja00870a028RIGHTS & PERMISSIONSArticle Views47Altmetric-Citations41LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit PDF (1 MB) Get e-Alerts Get e-Alerts

Site of cleavage of γ-globulins by papain

Limited cleavage of γ-globulins of several species by a variety of proteolytic enzymes yields 3.5-S fragments apparently representing adjoining structural moieties of the parent molecules. The locus, specificity, and stoichiometry of the peptide bonds involved have been studied by N-terminal group assay. Sulfhydryl-activated plant proteases such as papain, bromelain, ficin, and chymopapain all cleave similar bonds in human γ-globulin and form similar 3.5-S fragments. The γ-globulins of different species vary in susceptibility to papain; rabbit γ-globulin and pathological human proteins are most readily cleaved. Ficin and papain yield indistinguishable fragments from rabbit γ-globulin, namely Fractions I and II, and Fraction III. The latter can be resolved into three subfractions, IIIA, B, and C. In general, the sensitive bond involves a branched-chain amino acid, usually leucine. However, a transient threonyl N-terminal group is liberated in early stages of the cleavage of normal γ-globulin later being replaced by leucine. Both peptide-bond rupture and disulfide-bond reduction are requisite for the formation of 3.5-S fragments, but the cleavage is apparently initiated by the proteolysis which exposes a critical disulfide bond. However, no structure common to the γ-globulins of the various species can yet be formulated because they differ in the nature and apparent number of N-terminal groups and do not yield the same number or types of 3.5-S fragments on cleavage by papain.

Gm Factors in Normal γ-Globulin Fractions, Myeloma Proteins, and Microglobulins

The Gm groups in man are genetically determined properties of normal serum γ-globulins. In the present study, Gm factors (a), (b), and (x) were measured in fractions of normal serum γ-globulin obtained by anion-exchange (DEAE) cellulose chromatography. Gm factors appeared to be present in the same proportion in all the normal human γ-globulin fractions, with ultracentrifugal sedimentation coefficient of about 6.6S. However, the normal fraction, composed primarily of 18S globulin, had no detectable activity. Myeloma proteins, classified immunoelectrophoretically as γ-type and having a single 6.6S component in the ultra-centrifuge, varied in their Gm activities. Two Gm factors were undoubtedly present on one such protein. Other myeloma proteins appeared to have only one of the two or three Gm factors present in normal γ-globulins. Several myeloma proteins showed activity equal to that of normal γ-globulin for one Gm factor, but other Gm factors were found at very low levels. The quantitative differences in Gm activity of the individual γ-type myeloma globulins are believed to reflect qualitative differences in the proteins. Myeloma proteins classified immunoelectrophoretically as β2A-type and having multiple (6.6S, 9S, 11S, and 13S) components in the ultra-centrifuge did not have demonstrable Gm factors. The γ-18S macroglobulins (β2M on immunoelectrophoresis) similarly were without Gm factors. Reduction of these macromolecules to smaller units with sedimentation coefficients of about 6.6S did not uncover masked or latent Gm activity. The finding of Gm activity on γ-myeloma globulins indicates that malignant plasma cells retain certain genetic features of normal plasma cells.

On the associated state of rabbit allotypes, the existence of rabbit antibody molecules against two allotypes, and the dissociation of human γ-globulin antigens into smaller molecules

This chapter discusses the physical, chemical, and biological aspects of γM antibodies in depth. The chapter describes the physical characteristics, chemical composition, and subunit structure of the typical circular γM pentomers and the relationship of these molecules to low molecular weight γM-like proteins. The characteristics of the interaction of γM antibodies with antigens—for example, the nature and number of antigen combining sites—and the interaction of γM antibodies with the complement system are compared to the corresponding properties of γG antibodies. The biosynthesis and metabolism of γM antibodies and their peculiar role in the immune response are also discussed in the chapter. The chapter presents a brief summary related to the hallmarks of immunoglobulin heterogeneity and the criteria that must be used to judge the restricted heterogeneity or molecular uniformity of an antibody preparation. The chapter discusses human antibodies to certain selected antigens that appear to possess less heterogeneity than normal γ-globulin.