Γ2 Globulins Research Articles

F-Actin-Depolymerizing Activity of Human Serum

Non-heated human and animal sera contain a factor which exhibited an inhibiting activity on the staining of actin-containing structures by anti-actin antibodies in indirect immunofluorescence experiments. The presence of this factor lowered the viscosity of F-actin preparations and caused, as studied by electron-microscopy, a depolymerization of F-actin filaments as well as inhibition of filament formation of G-actin. The factor was, after its reaction with F-actin, liberated seemingly unaffected, indicating an enzymatic activity. The factor tentatively termed 'F-actin depolymerizing factor' was heat-sensitive and trypsin sensitive but resisted reduction. It was Ca2+ dependent and the staining inhibiting reaction was faster at 30 degrees C and 37 degrees C than at lower temperatures. Gel filtration experiments on Sephadex G-200 suggested a molecular size of the actin depolymerizing factor slightly higher than that of albumin. The electrophoretic mobility was that of gamma 2 globulin. The physiological role of the factor might be to prevent the presence of F-actin filaments within the circulation.

Studies on .GAMMA. globulin of rice embryo. VII. Localization of .GAMMA. globulin in rice seed and changes in .GAMMA. globulin content during seed development and germination.

Three components of γ globulins, γ1 γ2 and γ3, in rice seed were investigated immunochemically. γ1 Globulin was found to be immunochemically distinct from γ3 globulin because the immuno-precipitin arcs crossed, while γ2 globulin was immunochemically identical with γ3 globulin as γ2 globulin reacted with the anti-γ3 globulin-serum. It was revealed by means of fluorescent-antibody technique that both γ1 and γ3 globulins were localized in scutellum and aleurone cells. The content of γ1 globulin in aleurone cells was higher than that in scutellum, while the relative contents of γ3 globulin in these tissues were in the reverse relation. During seed development, γ1 and γ3 globulins increased almost linearly from the 5th day to the 40th day after flowering. On the other hand, the amounts of γ1 and γ3 globulins decreased rapidly in the process of germination. The rate of disappearance of γ3 globulin was greater than that of γ1 globulin.

Suppression of rabbit ear skin homograft rejections with sheep anti-thymus globulins. A preliminary report.

AbstractTwo electrophorectically distinct “7s” γ1, and γ2 globulins were isolated from sheep anti‐rabbit thymus serum. The γ1 fraction was effective in prolonging skin homograft retention in rabbits but did not suppress humoral antibody formation. The γ2 fraction appears to contain other antibodies which react with rabbit tissue to cause anaphylactic‐like reactions.

Activation of Complement by Endotoxin: A Role for γ2 Globulin, C1, C4 and C2 in the Consumption of Terminal Complement Components by Endotoxin-Coated Erythrocytes

Abstract Sheep erythrocytes (E) coated with endotoxin (LPS) from Salmonella typhosa or Veillonella alcalescens (E-LPS) have been employed to define more precisely the nature of a pathway by which LPS activates the terminal components of complement (C). E-LPS consumed C in normal guinea pig serum in a manner identical to free LPS, that is, with marked consumption of C3 through C9 and little utilization of C1, C4 and C2. A γ2-migrating globulin, isolated from normal guinea pig serum by DEAE-cellulose chromatography, was required for C-mediated lysis of E-LPS in diluted guinea pig serum. Absorption experiments with E-LPS indicated that the heat stable γ2 globulin showed specificity for the LPS employed. Cobra venom factor, known to activate C by the alternate pathway (C3 proactivator system), was equally able to deplete C activity in normal serum or serum absorbed with E-LPS. E-LPS reacted with γ2 globulin, C1, C4, C2, followed by C-EDTA resulted in lysis of E-LPS. Deletion of γ2 globulin or any of the earlier acting C components from the reaction prevented lysis. The small amounts of the early acting C components employed, particularly C4 and C2, indicated that it would be difficult to measure their loss in whole serum after activation with LPS by conventional hemolytic assay. This could explain the finding of a relative sparing of C1, C4 and C2 during consumption of C3 through C9 by LPS in normal guinea pig serum. Thus, a classical C pathway involving a “natural” γ2 globulin and early components of C has been defined as a mechanism by which endotoxin can efficiently activate the terminal components of C.

Biochemistry, Schizophrenias, and Affective Illness

Reading this book is a frustrating experience. It is probably the best source of authoritative knowledge about the biochemistry of mental illness. It is encyclopedic in scope. Yet it leaves the clinician feeling dissatisfied. Why should this be? The editor, Dr. Himwich, has admirably described the present status of indoleamines in schizophrenia and depression. Drs. Snezhevsky and Vartanyan have reviewed the role of erythrocyte permeability in schizophrenia and given us the Russian approach to classification. Dr. Schildkraut has made an exhaustive, scholarly review of catecholamine metabolism and affective illness. Dr. Gottlieb and his co-workers have reviewed their provocative hypothesis about the role of γ2 globulin in schizophrenia and have suggested that this substance is out of control in the schizophrenic just as glucose is out of control in the diabetic. Dr. Heath has brought up to date his immunological theory of the etiology of schizophrenia and the possibility of an

Investigations on eosinophilia. The influence of histamine, antigen-antibody complexes containing gamma-1 or gamma-2 globulins, foreign bodies (phagocytosis) and disrupted mast cells.

SUMMARY.— Eosinophilia in guinea‐pigs and mice is mediated by antibodies that sensitize tissues anaphylactically. In the guinea‐pig these are guinea‐pig γ1 globulin antibodies and rabbit passive sensitizing antibodies; in the mouse, mouse γ1 globulin and reagin. The γ2 globulin antibodies in both species, mediate Arthus reactions without eosinophilia. However, in vitro, there is no difference between eosinophils and neutrophils in their response to antigen‐antibody complexes containing either γ2 or γ1 globulin, which are ingested by both types of cell. Eosinophils in anaphylactic or parasitized tissues are particularly attracted to large, basophilic mononuclear cells which are found in mesenteric milk spots, around parasitic granulomata and probably in lymph nodes. Histamine fails to evoke eosinophilia in guinea‐pigs and rats, though it traps eosinophils in the tissues, forming an apparent eosinophilia. Anti‐histamine drugs fail to suppress the eosinophilia of anaphylaxis in guinea‐pigs. Though generalized or local anaphylaxis results in eosinophilia, it is not known whether complexes of antigen with tissue passive sensitizing antibody on the cell surface attract the eosinophils, or whether they are attracted by some vascular permeability factor released from the anaphylactic cells.

The Specificity of the Early Immune Response

Abstract The specificity of circulating antibody and delayed hypersensitivity (DHS) was studied in both normal and Fab, Fc tolerant guinea pigs immunized with human γG globulin (HGG). During the first 2 weeks after immunization, between 88% and 100% of the circulating antibody in both normal and Fab, Fc tolerant animals was found to be specific for the intact HGG molecule and did not react with Fab or Fc fragments. By the 6th week, antibody of this specificity accounted for about 60% of the total antibody in the tolerant animals but only about 15% in normal animals. Such antibody was not associated with any particular immunoglobulin class, activity being demonstrated by radioimmunoelectrophoretic and antigen-binding studies to occur in both γ1 and γ2 globulin fractions. Antibody against Fab and Fc fragments was not demonstrable in sera from normal animals until the 3rd week following immunization, but in later sera accounted for about 80% of the total antibody production. Little or no antibody against such fragments was demonstrable in the sera of the tolerant animals. Skin testing with HGG, Fab and Fc fragments elicited comparable areas of erythema in normal animals over a wide range of antigen doses on several occasions following immunization. Fab, Fc tolerant animals on the other hand, gave DHS reactions only with the intact HGG molecule. There thus appears to be a difference in the specificity of humoral and cell mediated responses to HGG, particularly in the first 2 weeks after immunization. It is considered possible that the earliest humoral response might, as a general phenomenon, have a specificity directed against the intact configurational form of an antigen.

Increased IgG synthesis during the induction of immunological paralysis in adult guinea pigs.

IT is well known that after immunological stimulation, animals produce non-specific gamma globulins as well as specific antibody1. Quantitative studies have shown that injection into guinea-pigs of an antigen mixed with Freund's adjuvant causes a marked increase in the serum concentrations of γ1 and γ2 globulins, only part of which can precipitate with the antigen2. On the other hand, the induction of immune paralysis in adult guinea-pigs is accompanied by a transient immunological unresponsiveness to unrelated antigens3. A possible explanation for this phenomenon could be that immunological paralysis may start by a defective commitment of immunologically competent cells, which does not result in the production of specific antibody. The aim of the present work was to find out whether the induction of paralysis also results in an increase of γ1 and γ2 globulins which might account for some kind of commitment of the competent cells.

Strain variations of mouse Sγ1 globulins

Strain variations in the migration of the S γ1 globulin of mice have been demonstrated by immunoelectrophoresis. The variation of electrophoretic mobility seems to have a hereditary basis and was not accompanied by any detectable antigenic dissimilarity. The fast migration of the CBA 7 S γ1 was presumably not due to to the presence of sialic acid. Papain digestion of the 7 S γ1 from a strain of mice with 7 S γ1 of fast mobility (CBA) and a strain with slow mobility (C57BL/6) showed that both the F c and the F ab pieces migrated at rate similar to the parent molecule. In addition, the F c piece of the 7 S γ1 was seen to migrate faster than the F c piece of the 7 S γ2 globulin.

Hexose Content of Guinea Pig 1 and 2 Immunoglobulins.

SummaryVarious preparations of normal and antibody guinea pig γ1 and γ2 globulins have been analyzed for their hexose content, using the indol method. The value obtained, when expressed in glucose equivalents, was 0.74% for both types of γ-globulin. Analysis of the different fragments obtained by enzymatic splitting of the molecule of γ2 globulin indicated that most of the hexose was contained in the fast (F) fragment obtained after digestion with papain (piece III of Porter). The fragment obtained after pepsin digestion contained very little if any hexose. The results indicate that guinea pig γ1 globulin does not correspond to the γ1A globulin of man and mouse. It is suggested that γ1 globulin represents a distinct family of im-munoglobulins, possibly also existent in other animal species, which mediates anaphylactic reactions within the species.

ANTIBODIES IN HUMAN COCCIDIOIDOMYCOSIS: IMMUNOELECTROPHORETIC PROPERTIES.

SummaryAn immunoelectrophoretic study was undertaken on two types of antibody detectable in human coccidioidomycosis, early precipitins and later complement-fixing antibodies. By using concentrated precipitin sera and dilute coccidioidal antigen, arcs of precipitation were produced which appeared to correspond to the mobility of 19S β2 macro-globulin. Complement-fixing sera and cerebro-spinal fluids reacted with coccidioidal antigen to give precipitation arcs with the mobility of 7S γ2 globulin.

EFFECT OF ALDEHYDR FIXATION ON CELLULAR RHEUMATOID FACTOR AND CERTAIN TISSUE ANTIGENS.

Rheumatoid facor, nuclei, and γ2 globulins can be identified by immunofluorescence microscopy in alcohol-fixed paraffin-embedded sections, though wax of low melting point is required for preservation of nuclear antigens. Rheumatoid factor also resisted aldehyde fixatives; hence it should be possible to investigate the ultrastructural characteristics of cells containing it.

CUTANEOUS ANAPHYLACTIC REACTIONS WITH ANTIBODIES OF DIFFERENT QUALITIES.

These results indicate that a 7S γ2 rabbit antibody having a low affinity for its antigen was equally as efficient as a 7S γ2 high affinity antibody in producing cutaneous anaphylaxis in guinea pigs when abundant antigen was present. However, when the concentration of antigen in the reaction was reduced the antibody of low affinity was not as potent as the antibody of higher affinity. A possible explanation of the inefficiency, marked by a slow rate of development and at times less severe anaphylactic reaction, was found to lie in a relatively slower rate of association noted between the absorbed antiserum and its antigen (Fig. 1). This slower rate of association may well bear causal significance when considering the finding that when one merely increases the concentration of antigen in the circulation, the rate of anaphylactic response of the slowly associating antiserum was increased to equal that of the control, rapidly associating antiserum (Table I). These latter conditions favored more rapid and complete binding of the lower affinity antiserum by antigen. These findings fail to support the possibility that the absorbed, slower associating antibody was less capable of “fixing” to tissues or for other reasons less capable of reacting, since merely by raising the dosage of antigen it became fully reactive in vivo. Other tests failed to show significant differences between antibodies in the 2 antisera: both were 7S, γ2 globulins as determined by density gradient ultracentrifugation and radioimmunoelectro-phoresis. When mixed with antigen they both fixed abundant hemolytic complement. The antigen-antibody dissociation rates of the 2 antisera were not appreciably different considering the amount of time required to demonstrate as much as 15-20% difference. Further, in other unpublished studies in this laboratory, results similar to those herein reported have been found using anti BSA obtained from individual rabbits after primary immunization and absorbed once with a small amount of antigen. This diminishes the possibility that the results could be explained by some unique quality of the absorbed antiserum.

THE PROTEINS OF EEL SERUM: STARCH GEL ELECTROPHORESIS AND IMMUNOLOGICAL STUDIES.

SummaryEel serum (Anguilla vulgaris) has been examined using several electrophoretic technics such as paper, agar gel, starch gel, and double electrophoresis, and using immunological studies such as immunoelectrophoresis and the Ouchterlony diffusion test. Twenty-one components were observed. No γ2 globulin was seen in the normal sera but it appears after immunization of eel against human proteins. The serum of eels contains at least 12 immunologically different antigens to rabbits. We observed most individual electrophoretic differences in the β region of the protein pattern.

The preparation and uses of antiglobulin reagents with special reference to complement-fixing blood group antibodies.

This paper describes the preparation of anti‐C′, anti‐γ1 and anti‐γ2 antiglobulin reagents especially for use with human cells sensitized with blood group antibodies. Anti‐γ1 and anti‐γ2 are carefully prepared so as to be as specifically active as possible. The paper also describes the value and application of these reagents in clinical diagnostic work and their use in the detection of γ1 and γ2 globulins in solution, or as they occur in the form of antibody absorbed on to erythrocytes. By the use of these sera it is possible to determine the nature of the antibody coating sensitized cells. Within the limitations described, it is possible to do this with the cells coated not only with antibody but with complement as well.The value of these sera in experimental work, with particular reference to the demonstration of the action of human complement on red cells sensitized with blood group antibodies, is described; it is shown that the action between sensitized cells and anti‐γ1 antiglobulin serum is depressed by complement activity and the significance of these observations is discussed.

Studies of blood group antibodies, V. Fractionation of examples of anti-B, anti-A,B, anti-P, anti-Jka, anti-Lea, anti-D, anti-CD, anti-K, anti-Fya, anti-s and anti-Good.

Examples of blood group antibodies were subjected to density gradient ultracentrifugation and anion‐cation chromatography on modified cellulose adsorbents and three fractions were collected. The first fraction owed its serologic activity to the presence of γ2 globulins with a 7S sedimentation constant; the principal serologically‐active component in fraction two was γ1 globulin of the 7S class; in most cases the serologically‐active component of the third fraction was γ1 globulin of the 19S class. Two anti‐A donors were found to have most of the serologic activity in fraction three before injection of blood group substance and a marked rise in fractions two and three after stimulation. In anti‐M, anti‐P and saline anti‐D sera the serologic activity associated with S19 globulins equalled or exceeded the activity with the S7 globulins. Group O serums were studied before injection of A and B substances and S7 activity was preponderant in 48 per cent whereas post‐immunization it was preponderant in 88 per cent. Fraction one activity was also preponderant in examples of anti‐K, anti‐Fya, anti‐s, anti‐Jka and anti‐Good. Those antibodies which are most likely to cause erythroblastosis fetalis are those which are characterized by a predominance of the S7, γ2 component.

Study of Bence-Jones proteins

SummaryA biochemical study of fourteen cases of multiple myeloma with Bence-Jones proteins present in the urine, is made. Bence-Jones proteins have several properties in common which distinguish them from the other normal or abnormal serum proteins. Thanks to their low molecular weight, they pass through the kidney and can be easily characterised in and extracted from the urine. Their electrophoretic mobilities are situated between those of the β and γ2 globulins in the serum.Complete thermosolubility at 100° C is not an absolute criterium for their identification. The presence of non-soluble fractions at boiling point does not necessarily imply the presence of other proteins.The amino acid content of a given Bence-Jones protein as determined by Moore and Stein’s method does not seem to differ appreciably according to the process used to extract it from the urine.The comparison of our results concerning the amino acid composition of electrophoretically differing Bence-Jones proteins shows an appreciable d...