bluuebunny/crossref_metadata_2025 huggingface dataset:
Cyclophosphamide Inhibition of Experimental Allergic Encephalomyelitis and Cellular Transfer of the Disease in Lewis Rats
Authors: Paterson Philip Y. , Hanson Margaret A.
Published: December 2022

Single or daily injections of cyclophosphamide (CY) at appropriate dose levels completely inhibit the development of experimental allergic encephalomyelitis (EAE) in Lewis rats sensitized to spinal cord in adjuvant. CY treatment of sensitized Lewis donors or injection of CY into recipients after injection of donor lymphoid cells suppresses or inhibits transfer of EAE. Treatment of prospective recipients before injection of donor cells also may inhibit transfer. These observations, together with other data cited, suggest that active immunologic participation of the recipient may be a prerequisite for successful transfer of the disease.
Quantitative and Qualitative Studies on the Primary Antibody Response to Pneumococcal Polysaccharides at the Cellular Level
Authors: Baker Phillip J. , Stashak Philip W.
Published: December 2022

The kinetics of the antibody response to purified pneumococcal polysaccharides was studied at the cellular level using the technique of localized hemolysis-in-gel in conjunction with erythrocytes sensitized with immunizing antigen. The overall pattern of the antibody response was similar, with respect to the number of direct plaqueforming cells produced, to that reported for other antigens. However, using antisera specific for various classes of mouse immunoglobulins to characterize the types of specific plaque-forming cells produced, it appears that the antibody response to pneumococcal polysaccharides is exclusively macroglobulin in nature; two types of macroglobulin antibodies, differentiated on the basis of their efficiency in participating in immune hemolysis, are produced. The data suggest that direct plaque-forming cells may represent only 30% to 50% of the total number of antibody-forming cells produced following immunization.
Isolation and Characterization of IgA Produced by an Established Human Lymphocytoid Cell Line
Authors: Matsuoka Y. , Yagi Y. , Moore G. E. , Pressman D.
Published: December 2022

Extracellular and intracellular human immunoglobulins of the IgA class were isolated in a highly purified state from the culture of an established human lymphocytoid cell line RPMI 4666. It was found that the general features of these immunoglobulins are similar to the serum IgA in myeloma individuals, consisting of covalently-bound α heavy chains and kappa light chains and possessing idiotypic antigenic determinants in addition to the antigenic determinants common to other κ type IgA. The extracellular immunoglobulin was found in both monomeric and dimeric forms while the intracellular immunoglobulin was found only in the monomeric form. Although the extracellular and intracellular immunoglobulins differed distinctly in their electrophoretic mobility, no difference could be detected in their antigenic determinants. The difference in their mobilities was shown to be reflected in their Fab-fragments.
Mode of Action of Adjuvants: Effects on Antibody Responses to Macrophage-Associated Bovine Serum Albumin
Authors: Spitznagel John K. , Allison A. C.
Published: December 2022

Primary and secondary antibody responses were measured in CBA mice after injecting them with different primary doses of free bovine serum albumin (free BSA) or BSA taken up by syngeneic macrophages (MBSA). The antibody responses to MBSA were much higher than those to FBSA, and the slopes of the dose-response curves differed significantly. When FBSA was administered shortly before MBSA, the antibody responses were less than when MBSA was given alone. It is suggested that FBSA elicits two competitive responses in immunocompetent cells: immunization and the induction of tolerance; with MBSA the balance is changed strongly toward immunization.

Potent adjuvants, injected prior to FBSA, heightened antibody responses to approximately the level produced by the same doses of MBSA. This suggested that adjuvants can also prevent the tolerance-inducing effect of FBSA. MBSA is retained by macrophages incubated in vitro for several hours, and the immunogenicity of the retained BSA does not decrease. MBSA gives rise to an antibody response in irradiated recipients only if they have been reconstituted with syngeneic lymphocytes. MBSA in E. coli lipopolysaccharide (LPS)-treated macrophages induced consistently higher antibody responses in normal mice than MBSA bound by untreated macrophages. LPS treatment of lymphocytes used to reconstitute irradiated animals had no detectable effect on antibody responses. This suggests that the effects of adjuvants may be exerted at least partly on macrophages. LPS had no demonstrable effect on the uptake or degradation of antigen by macrophages.
Binding Properties of Anti-Dinitrophenylated-Bacitracin A Antibodies Produced in Guinea Pigs
Authors: Mancino Domenico , Paul William E. , Benacerraf Baruj , Siskind Gregory W. , Ovary Zoltan
Published: December 2022

Guinea pigs immunized with tri-DNP-bacitracin A produced anti-DNP-antibodies with binding characteristics which varied from individual to individual. Of nine antisera from single animals studied by equilibrium dialysis against tritiated DNP-ε-aminocaproic acid (DNP-EACA), DNP-δ-aminovaleric acid (DNP-DAVA) and DNP-isoleucine (DNP-ile), three bound DNP-EACA and DNP-DAVA clearly better than they bound DNP-ile, one bound DNP-ile with a significantly greater energy of interaction that it bound DNP-DAVA, and five bound the ligands with similar or only slightly different energies of interaction. In contrast, anti-DNP antisera produced by seven individual guinea pigs immunized with DNP-BGG or DNP-BSA all bound DNP-EACA or DNP-DAVA to a greater degree than they bound DNP-ile. These observations suggest that individual animals can respond preferentially to one of a series of antigenic determinants presented on the same molecule.
Demonstration of Viral Antibody Activity in Two Immunoglobulin G Subclasses in Patients with Varicella-Zoster Virus Infection
Authors: Leonard Larry L. , Schmidt Nathalie J. , Lennette Edwin H.
Published: December 2022

The nature of the immunoglobulins produced in response to infection with the varicella-zoster virus (V-Z) was investigated. In primary infections (varicella) antibody was demonstrable in both IgM and IgG serum fractions, while in secondary infections (herpes zoster) antibody was demonstrable only in the IgG fraction. Two IgG components differing in their electrophoretic mobilities were separated from human sera by DEAE-cellulose chromatography and these differed in their antibody activity depending upon the clinical manifestation of the V-Z infection. V-Z antibody was demonstrable in the “slow” IgG subclass in varicella and herpes zoster infections by both neutralization and fluorescent antibody (FA) tests. In varicella infections neutralizing antibody was not demonstrable in the “fast” IgG fraction, although low levels of antibody were detected by FA tests. In herpes zoster infections, on the other hand, the “fast” IgG fraction contained antibody demonstrable by both neutralization and FA tests. This suggests that the antibody response to primary infection with the V-Z may be “incomplete.” The possible protective activities of these two subclasses of IgG are discussed.
Human M-Proteins with Antibody Activity for Nitrophenyl Ligands
Authors: Terry W. D. , Boyd M. M. , Rea J. S. , Stein R.
Published: December 2022

Human sera containing M-proteins were tested for their capacity to bind nitrophenyl haptens. About 1.5% of 580 M-proteins showed significant binding for either DNP-caproate or DNP-lysine. In all active sera, binding was due to the M-protein. By contrast, tests of 319 sera by immunodiffusion against nitrophenyl-protein conjugates resulted in many false positive and false negative reactions, indicating that this is a less satisfactory method for detecting M-proteins with antibody activity.
Pyrrolidonecarboxylic Acid at the N-Terminal Positions of Polypeptide Chains from Leopard Shark Immunoglobulins
Authors: Goodman Joel W. , Klaus Gerhard G. , Nitecki Danute E. , Wang An-Chuan
Published: December 2022

Leopard shark immunoglobulins are found in 7S and 17S forms, which are indistinguishable chemically and antigenically, and probably are analogous to IgM molecules of mammals (1). The two molecular forms thus appear to share common H and L polypeptide chains. Suran and Papermaster reported that the amino terminal 6 residues from H- and L-chains of at least a portion of 7S and 17S leopard shark Ig were virtually identical to each other and to the homologous regions of kappa L-chains of mouse and man (2). These results suggest that relatively little divergent evolution had occurred in this region of H- and L-chains at the level of the elasmobranchs and support the common origin of these chains by duplication of an ancestral gene (3, 4). However, lambda L-chains and the gamma and mu H-chains from mammalian Ig almost invariably possess a blocked N-terminal residue identified as pyrrolidonecarboxylic acid (PCA).
Concanavalin A-Induced Release of Skin-Reactive Factor from Lymphoid Cells
Authors: Schwartz Howard J. , Leon Myron A. , Pelley Ronald P.
Published: December 2022

We have found that pretreatment of tuberculin-sensitive guinea pigs with the plant lectin concanavalin A suppresses their delayed hypersensitivity response (1). Concanavalin A also complexes with glycoproteins, and possibly glycolipids, at the surface of a variety of cells (2, 3), and with glycoproteins in body fluids (4, 5). In the case of lymphocytes, this binding initiates a series of reactions leading to blast transformation (3). The observation that phytohemagglutinin (another plant lectin) activates lymphocytes for the production of lymphotoxin (7–9) prompted us to further examine the effect of concanavalin A on normal lymphocytes.

Concanavalin A was prepared by a minor modification (5) of the method of Agrawal and Goldstein (6). Axillary and inguinal lymph nodes and spleens were obtained from non-immune, randomly bred albino guinea pigs following exsanguination by cardiac puncture. The organs were diced and teased gently with mouse-toothed forceps in cold Hanks' balanced salt solution and then filtered through sterile glass wool twice.
Bacteriologic Studies on Wasting Disease Induced by Neonatal Thymectomy
Authors: Cantrell John C. , Jutila John W.
Published: December 2022

Neonatal Balb/c mice thymectomized at 12 to 24 hours of age and treated with two injections of rabbit anti-mouse thymocyte serum (RAMTS) developed a more severe wasting disease with a more rapid onset than did their thymectomized counterparts. Neonatal Balb/c mice that received only RAMTS failed to demonstrate symptoms of wasting disease.

The liver, lungs, spleen and blood of thymectomized Balb/c neonatal mice treated with RAMTS were found to be infected with several species of microorganisms presumed to originate from the normal flora of the gut. In the early onset of infection, a nonpathogenic species of Erysipelothrix was the predominant organism cultured from the tissues. As time progressed, mixed infections consisting of Streptococcus faecalis, Corynebacterium species, Bacillus species and Proteus mirabilis were commonly found in the tissues and organs of RAMTS-treated thymectomized mice. Citrobacter species and S. faecalis were the predominant microorganisms isolated from mice that showed terminal wasting disease.
Development and Duration of Mumps Fluorescent Antibodies in Various Immunoglobulin Fractions of Human Serum
Authors: Brown Gordon C. , Baublis Joseph V. , O'Leary Theresa P.
Published: December 2022

The indirect immunofluorescence technique was successfully adapted to the detection and titration of mumps virus antibodies in human sera. The curve of antibody development and duration for 3 months following the onset of clinical disease was measured with fluorescein-labeled antisera prepared against the IgG, IgA and IgM fractions as well as the whole human GG, and was compared with neutralizing antibodies. Antibodies in the whole GG and the IgG fraction were present in high titer at the time of onset and persisted unchanged for at least 3 months. IgM antibodies also developed rapidly to somewhat lower titers but declined steadily and were rarely detectable 50 days after the disease. Limited observations on the curve of IgA antibodies indicated that they too are transient and decline during the first month after infection. Neutralizing antibodies in these same individuals developed slowly and to relatively lower titers but persisted for the duration of the study. The specificity of reactions with labeled antisera for various fractions of GG was confirmed by treatment with 2-ME followed by density gradient centrifugation.

The FA response of individuals receiving live, attenuated mumps virus vaccine was measured using conjugates prepared against IgM and whole GG and was found to correspond closely to that measured by the neutralization technique.

The extent of subclinical infection and immunity among 48 ward contacts of cases in an institution was determined by testing paired serum specimens which had been collected at an interval of 36 days. Thirty-one per cent were found to be serologically immune. Twenty-five per cent remained susceptible and 43% experienced subclinical infection.

Reactions between antibodies in sera of patients convalescent from mumps and antigens prepared with parainfluenza viruses of types 1, 2 and 3 were prominent with fluorescein-labeled antisera against whole GG but were completely eliminated when anti-IgM serum was employed. This rapid and highly specific diagnostic procedure has the added significance that a positive reaction in even a single serum specimen proves that the individual has experienced infection during the past few weeks.
Inhibition of the Eighth Component of Complement (C8) by Ethylenediaminetetra-Acetate (Edta)
Authors: Schultz Duane R. , Zarco Romeo M.
Published: December 2022

Functionally pure C8 from human or guinea pig serum was inactivated by 1 to 2 × 10-3 M EDTA. Native C8 in serum was partially inactivated by EDTA, but much longer periods of incubation at 37°C were required for this to be evident. The effect of EDTA on C8 was dependent upon time, temperature, and concentrations of both the chelating agent and C8. The inactivation of C8 by EDTA was partially or totally reversed by the addition of Ca2+ or by dialysis, but reversal was dependent upon the duration of exposure and the temperature. The process became irreversible within relatively short periods of time at 30°C and 37°C. EDTA had no apparent effect on the activity of cell-bound C8 or on the binding site of C8 for C9, but it did impair the binding site of C8 for C7. Neither EAC1–7 nor C9 was affected by EDTA.
The Biologic Activities of Guinea Pig Antibodies
Authors: Oliveira Benedito , Osler Abraham G. , Siraganian Reuben P. , Sandberg Ann L.
Published: December 2022

Guinea pig immunoglobulins have been separated into the γ1 and γ2 classes by chromatography on DEAE cellulose. Both types show charge heterogeneity. The heterogeneity of the γ2 immunoglobulins is attributed to the F(ab) fragments. As noted previously by others, the ability to mediate passive cutaneous anaphylaxis (PCA) in the guinea pig skin is confined almost entirely to the γ1 immunoglobulins. Both types of immunoglobulins participate in C-dependent allergic reactions of the immediate type. These include the generation of anaphylatoxin, the mediation of reverse passive Arthus reactions, as well as allergic histamine release from normal rabbit platelets.
Synthesis of Secretory IgA in the Rabbit
Authors: Lawton Alexander R. , Asofsky Richard , Mage Rose G.
Published: December 2022

Rabbit colostral IgA internally labeled with 14C has been isolated from culture fluids of short term cultures of mammary tissue obtained from three rabbits. Alpha, light, and T chains of intact or extensively reduced and alkylated IgA were separated by Sephadex G-200 chromatography in 5 M guanidine, and by polyacrylamide disc electrophoresis. The results indicate that all three chains are synthesized locally, and that 14C lysine and isoleucine are incorporated into α, light and T chains in a ratio of approximately 4:2:1. This ratio is in reasonably close agreement with the mass ratio of the chains of an IgA molecule with 4α, 4 light, and 2 T chains. The data indicate that local synthesis can account for all of the IgA found in rabbit colostrum.
Antigenic Specificity of γA Polymers
Authors: Apicella Michael A. , Allen James C.
Published: December 2022

Studies of the polymer and monomer γA isolated from a single patient with γA multiple myeloma indicate that the polymer has an antigenic specificity distinct from the monomer. This specificity is dependent upon the polymer configuration and is unrelated to individual antigenic specificity.
A New Serum Factor in Normal Rabbits
Authors: Woolsey Marion E. , Mandy William J.
Published: December 2022

The levels of HR activity in normal adult rabbits were not affected by immunization with autologous intact γG-globulin or with the purified Fab and Fab′ fragments of autologous γG-globulin.

The effect of immunizing newborn rabbits with homologous γG-globulin or with the enzymatic fragments of homologous γG-globulin on HR synthesis was demonstrated by an increase in the levels of HR significantly beyond the levels found in control rabbits of the same litter at the same age.

The induced HR was similar to the naturally-occurring HR in: a) specificity for buried determinants of homologous γG-globulin, and b) specificity for either Fab or Fab′ fragments as the provoking antigens. Of particular interest was the finding that HR for both the Fab and Fab′ specificity was induced in newborn rabbits by immunization with intact γG-globulin.

These findings are offered as supportive evidence that HR synthesis is prompted and maintained by antigenic stimuli resulting from degradation products of autologous γG-globulin occurring as early as 4 to 5 weeks after parturition.
Immunoglobulin Classes of Antibody-Forming Cells in Mice
Authors: Nordin Albert A. , Cosenza Humberto , Sell Stewart
Published: December 2022

Spleen cell suspensions from mice that had been immunized with an optimal dose of sheep erythrocytes were assayed by a replicate plating technique capable of detecting and separately differentiating plaque-forming cells (PFC) producing IgM and/or γ1 hemolysin. The mice were tested 5 1/2 days after immunization when γ1 PFC were first detectable in significant numbers; approximately equal to the number of IgM PFC. Only 2% to 4% of the PFC examined could be considered as producing IgM and γ1 hemolysin simultaneously. These results are inconsistent with the theory that the rapid appearance of a large number of IgG-PFC between day 4 and day 51/2 is the result of antibody-producing cells switching from IgM to IgG production.
Communications
Authors: Nagaya H. , McKenzie W. N. , Kilburn K. H. , Sieker H. O.
Published: December 2022

We have shown that the immunosuppressive effect of antithymus serum is greater than that of antilymphocyte serum in rats (1, 2). Histologically, the thymus of animals treated with antithymus serum shows marked depletion of small lymphocytes whereas the thymus of rats treated with antilymphocyte serum appears to be intact. Our subsequent radioautographic studies indicated that thymic lymphocyte production was suppressed in rats receiving antithymus serum, but was increased in animals receiving antilymphocyte serum (3, 4). These findings suggest that suppression of thymic lymphocyte production is one of the important mechanisms of the immunosuppressive action of antithymus serum. This communication reports that in mice, also, antithymus serum has greater immunosuppressive potency than does antilymphocyte serum. Furthermore, thymus antigens responsible for the production of potent antiserum are closely associated with the ribosomal fraction of thymus glands.

Antithymus serum (ATS) was produced in rabbits by 5 weekly subcutaneous injections with a mixture (2.5 ml) of a 20% saline suspension (by volume) of 6- to 8-week-old AKR mice thymus homogenates and Freund's complete adjuvant.
Evidence for Conformational Unfolding of Immunoglobulin Heavy Chains in Propionic and Acetic Acids
Authors: Harrington Judson C. , Fenton John W.
Published: December 2022

The procedures of Fleischman et al. (1, 2) are widely used to prepare immunoglobulin heavy (H) and light (L) polypeptide chains. Because they reported higher yields of L-chains separated in 1 M propionic acid than in other solvents (1), propionic acid is traditionally used for gel-filtration fractionation of chains from reduced and alkylated immunoglobulins. When chains are separated on Sephadex G-75 in this solvent, two peaks are characteristically obtained which correspond to the H- and L-chain fractions, respectively. However, when chains are similarly separated on higher resolving Sephadex grades (i.e., G-100, G-150, or G-200), three peaks are generally obtained (3–5). Here the initial peak is commonly referred to as the X fraction and is followed by the H- and L-chain fractions. Because the X peak is not apparent in profiles of chains separated in protein dissociating solvents, such as 5 M guanidine hydrochloride (6), 6 M urea (7), or detergent (8), the X fraction is presumed to represent an aggregated form of H-chains.
Model Systems for the Study of Drug Hypersensitivity
Authors: Hoffman Donald R. , Campbell Dan H.
Published: December 2022

Normal rabbit serum is acetylated by aspirin under in vitro conditions, which are similar to physiologic conditions. The extent of acetylation is determined by immunochemical tests. The acetylated serum proteins are shown to be potent antigens in rabbits. It is also shown that aspiryl proteins are partially acetylated. The relation of these results to human aspirin intolerance is discussed.
Late 19S Rabbit Antibody Neutralization Test for Differentiating Herpes Simplex Virus Types 1 and 2
Authors: Hampar Berge , Martos Lidia M. , Chakrabarty Mrinal , Burroughs Mary Alice K.
Published: December 2022

Early (7 day) and late (hyperimmune) sera were prepared in rabbits against two type 1 and one type 2 strains of herpes simplex virus (HSV), and neutralization tests were carried out. The results indicated that the late 19S antibodies were at least four to eight times more effective than were the early 7S and 19S antibodies or the late 7S antibodies in their ability to differentiate between HSV types 1 and 2. The potential role of late 19S rabbit neutralizing antibodies as herpesvirus typing reagents is discussed.
Enhancing Antibody
Authors: Zimmerman Barry , Feldman Joseph D.
Published: December 2022

Parenteral administration of Le and LBNF1 anti-Bf sera prolonged the MST of Bf grafts from 7.6 days to 16 and 14 days, respectively. IgG prepared from these sera were also biologically active but increased mean survival times to less than 13 days. Immersion of Bf grafts in anti-Bf whole serum or IgG also was effective in prolonging survival of Bf grafts. When two Bf grafts, one dipped in anti-Bf serum or IgG and the other undipped, were sutured on neonatal recipients, the dipped graft was enhanced in two-thirds of the trials while the undipped graft was rejected within control times. These results were interpreted to mean that enhancing antibodies were active peripherally either in the skin graft or its draining lymph node. Despite the use of isotopes, direct evidence for binding to the homograft was not obtained.
Reactions in Agarose Gel between Clq and Aggregated γ Globulin
Authors: Thunold S. , Abeyounis C. J. , Milgrom F.
Published: December 2022

The reaction of normal human and animal sera with heat-aggregated serum γ globulin (FII) was studied by means of double diffusion precipitation tests in agarose gel. Best results were obtained using agarose dissolved in a low molar salt solution (0.075 M NaCl). Precipitation lines were obtained, and in some instances improved, in the presence of EDTA. The double diffusion tests in agarose gel could be employed as a simple and convenient analytic tool for identification of the precipitin as the Clq component of complement.

The reaction of Clq of human and animal origin was stronger with aggregated rat FII than with aggregated human FII. No reactions were obtained with bovine, sheep or rabbit FII. The anticomplementary activity of FII preparations in a hemolytic system paralleled their activity in the precipitation tests.

Eluates containing Clq were prepared from precipitates formed in the reaction between human serum and rat FII in the presence of EDTA. Antisera specific for human Clq were produced in rats by injection of such precipitates.
A Study of Antibody Responses by Radioimmunodiffusion with Demonstration of γ D Antigen-Binding Activity in Four Sera
Authors: Heiner Douglas C. , Rose Bram
Published: December 2022

Sixty-six selected patients were studied using radioimmunodiffusion for evidence of antigen-binding activity by γD. Four were found to have γD antibodies, three of whom were sensitive to cow's milk and had γD antibodies directed toward bovine γ globulin. Two of these also had γD antibodies to bovine serum albumin, another constituent of cow's milk. In a fourth subject, γD antibodies were produced as a result of booster injections of diphtheria toxoid. Each of the subjects in whom γD antibodies were found had antigen-binding activity by other classes of immunoglobulins and had further evidence of vigorous immunologic responsiveness. Two had an antibody response to a specific antigen which involved all five immunoglobulin classes.
Tuberculin Sensitivity: The Effect of Antilymphocyte and Antimacrophage Serum on Cutaneous, Systemic and In Vitro Reactions
Authors: Heise E. R. , Weiser R. S.
Published: December 2022

Antilymphocyte serum (ALS) and antimacrophage serum (AMS) were employed in the tuberculin system in an attempt to define the role of lymphocytes and macrophages in the efferent limb of the delayed sensitivity response. Administration of ALS prior to or simultaneous with tuberculin challenge of BCG-sensitized guinea pigs suppressed both cutaneous and systemic reactivity. With few exceptions skin reactions were not completely suppressed nor were signs of systemic shock or lethality prevented in all cases. However, tuberculin skin reactions in all ALS-treated animals were markedly suppressed, as evidenced by a grossly apparent reduction in erythema and induration and by a lessened degree of cellular infiltration. Likewise, systemic tuberculin reactions were less frequent and occurred later in ALS-treated than in NRS-treated control animals.

Antimacrophage sera were much less effective than ALS in suppressing cutaneous tuberculin reactions. Whereas cutaneous reactions were only moderately suppressed in the AMS-treated group, systemic reactions were equal to or more severe in AMS-treated animals than they were in NRS-treated controls. It is unlikely that the difference between the suppressive effects of ALS and AMS was due to lack of specific cytotoxic activity of the AMS, since in vitro tests showed that the cytotoxic titer of the AMS for macrophages was greater than that of ALS for lymphocytes. That AMS was active in vivo was indicated by the finding that fewer alveolar macrophages were recovered from AMS-treated than from ALS- or NRS-treated guinea pigs. Attempts should be made to produce stronger and more specific AMS for further studies.

The capacity of ALS to modify systemic tuberculin shock has not been reported previously, but is consistent with the observation that sensitivity to systemic tuberculin shock can be transferred passively in the guinea pig with lymphoid cells but not with serum.
The Biologic Activities of Denatured Human IgG and its Heavy Chains
Authors: Hirose Shun-Ichi
Published: December 2022

tH (polymerized H-chain) of human IgG did not show skin-fixing activity in the guinea pig as judged by RPCA. The structures of pU-IgG (polymerized urea-treated IgG) and tH required for reactivity with rheumatoid factors are not related to the IgG site needed for skin fixation. tH does not increase vascular permeability. The results suggest that the skin-fixing site of IgG is dependent upon its three-dimensional structure and is easily lost by many modifications. On the other hand, the modified structure of IgG and H-chain exhibits reactivity with rheumatoid factors, but only non-alkylated, reduced and aggregated IgG increases vascular permeability. These results strongly suggest that newly-created conformational structures are essential for development of the ability to increase vascular permeability in the guinea pig skin and reactivity with rheumatoid factors, but that these structures are not identical.
Antibodies to Type III and Type VIII Pneumococcal Polysaccharides: Evidence for Restricted Structural Heterogeneity in Hyperimmunized Rabbits
Authors: Pincus Jack H. , Jaton Jean-Claude , Bloch Kurt J. , Haber Edgar
Published: December 2022

Two hundred and thirty-two rabbits were hyperimmunized with formalinized type III and type VIII pneumococci in an attempt to elicit the formation of structurally restricted antibody in high yield. Although precipitating antibody to either type III or tyep VIII pneumococcal polysaccharide could be demonstrated in the sera of all immunized rabbits, electrophoretic restriction of the antibody in the λ globulin region could be demonstrated only in a minority of the animals, e.g., in sera from 7% of rabbits immunized with tyep III and from 9% of rabbits injected with type VIII pneumococci. The concentration of type-specific antibody in serum of rabbits showing an electrophoretically-restricted λ globulin region ranged from 3.5 mg to 32 mg/ml of serum. For the isolation of antibodies to S3 or S8 in high yield, a specific immunoadsorbent synthesized from bromoacetylcellulose and a polysaccharide-bovine serum albumin conjugate was developed. Antibodies could be recovered in 75% to 85% yield after being eluted under mild conditions and were found to be 85% to 100% precipitable with the homologous polysaccharides.
Goldfish Immunoglobulins and Antibodies to Bovine Serum Albumin
Authors: Trump Gary N.
Published: December 2022

Whole goldfish anti-bovine serum albumin (BSA) serum was fractionated to yield two populations of antibody-containing immunoglobulins of different charge. Whole serum protein homologies of the two antibody preparations were demonstrated by Osserman-type immunoelectrophoretic tests. Diffusion-in-gel tests indicated that the two antibody populations shared at least one antigenic determinant, although one population was antigenically deficient with respect to the other. On the basis of their whole-serum homologies and antigenic disparities, the two antibody populations are thought to represent distinct serum proteins.
Relative Concentrations of IgM and IgG Antibodies during the Primary Response
Authors: LeFor W. M. , Bauer D. C.
Published: December 2022

The synthesis of IgM and IgG antibodies to bovine serum albumin was examined during the primary response in the rabbit. After separation of the two immunoglobulins by gel filtration or density gradient ultracentrifugation, the quantity of antibody was determined by a modified antigen-binding procedure. The assay allowed weight estimates of the antibody concentration to be made, was dependent only on the primary union of antigen with antibody and did not demonstrate a selective efficiency in detecting one immunoglobulin class of antibody over the other.

IgM and IgG antibodies initially appeared in the serum at approximately the same time. At the peak of the primary response IgG antibody was present in a 50-fold molar excess over IgM. Active synthesis of IgM antibody took place in the presence of this large excess of IgG antibody. These observations do not support an inhibitory role of IgG antibodies on IgM synthesis.

During the study it was determined that the intact IgM antibody molecule bound a maximum of five to six antigen molecules when measured by the modified antigen-binding procedure.
An Antigen Density Effect on the Hemolytic Efficiency of Complement
Authors: Linscott William D.
Published: December 2022

It has been generally accepted for many years that immune hemolysis is a “one-hit” phenomenon in that a single complement-fixing antibody site at the cell surface is sufficient for the complement (C) system to produce a local damaged site (S*) which will lead eventually to lysis of the cell (1). A single molecule of IgM antibody has been shown to be capable of initiating the lytic reaction, by fixing one molecule of the first component of C (C1) (2).

It has recently become apparent that not all target cells are equally susceptible to the cytolytic effect of IgM antibody and C, and that fixation of C1 at a sensitized site (SA) does not necessarily guarantee that the remaining C components will be able to convert the resulting SAC1 to S* and produce cytolysis; although this conversion efficiency is as high as 30% to 50% with sheep erythrocytes (E) and anti-Forssman antibody (3), it is only about 0.03% with IgM-sensitized nouse leukemia cells (4).
Tissue- and Species-Specific Antigens of Normal Human Prostatic Tissue
Authors: Ablin R. J. , Bronson Paul , Soanes Ward A. , Witebsky Ernest
Published: December 2022

Heteroimmunization of rabbits with saline extracts of normal human prostatic tissue incorporating complete Freund's adjuvant produced antibodies reacting with saline extracts of normal human prostatic tissue as well as with comparable extract preparations of other human tissues and fluids as judged by the criteria of double diffusion gel precipitation and tanned cell hemagglutination. Rigorous absorption studies of rabbit antisera to extracts of normal human prostatic tissue accompanied by enhanced resolution of lines of precipitation following incubation of gel diffusion plates in a buffer-substrate mixture incorporating lead nitrate and sodium-β-glycerophosphate revealed that a portion of these antibodies were prostate-specific, and in addition permitted identification of two prostatic tissue-specific antigens. By specific immunochemical staining of the gel precipitation pattern, one of the prostate tissue-specific antigens was identified as prostatic acid phosphatase.

Species-specificity studies of the antigens of the normal human prostate revealed that comparable antigens were absent from preparations of canine and rabbit prostatic tissue, but apparently shared to some degree with antigens of the cranial lobe of the monkey prostate.

Several sera of patients with benign and malignant diseases of the prostate gave precipitation reactions accompanied by staining for prostatic acid phosphatase, but the relevance of such reactions remains to be determined. In addition, preliminary gel diffusion precipitation studies suggested specific antigenic differences between normal, benign and malignant prostatic tissue extracts.

Immunoelectrophoretic analysis of extracts of normal human prostatic tissue and specifically absorbed homologous antisera revealed one arc of precipitation in the βγ-region of the electrophoretic field. Further resolution of immunoelectrophoretic patterns obtained by incubation in the buffer-substrate mixture and staining for acid phosphatase disclosed two to three additional lines of precipitation, the major arc of precipitation being identified as prostatic acid phosphatase.

The sensitivity and specificity of rabbit antisera to extracts of normal human prostatic tissue as evaluated by gel diffusion precipitation and immunoelectrophoresis, as well as the development of this immunologic response, was confirmed by both direct tanned cell hemagglutination and inhibition of tanned cell hemagglutination.
The Preparation of Human Kininogen and the Elicitation of Antibody for Use in a Radial Immuno-Diffusion Assay
Authors: Spragg Jocelyn , Haber Edgar , Austen K. Frank
Published: December 2022

Kininogen was purified from fresh human plasma using mild conditions and an inhibitor of the kinin-activating sequence. The material recovered after chromatography, gel filtration and electrophoresis was only of the low molecular weight variety. It has been shown to release kinin upon treatment with purified human plasma kallikrein, urinary kallikrein or trypsin.

Antibody elicited in rabbits with purified kininogen reacted with biologically active kininogen eluted from disc gel fractions. This antibody was employed in the preparation of a radial immunodiffusion assay for kininogen in human biologic fluids.
Transfer of Autoimmune Thyroiditis of the Rat with Lymph Node Cells
Authors: Twarog Frank J. , Rose Noel R.
Published: December 2022

Autoimmune thyroiditis was transferred within inbred strains of rats by lymph node cells. Cells obtained 7, 9 or 14 days after injection of donor animals were able to induce the thyroidal lesions. Rather larger numbers of cells were needed. Only 13% (4/31) of recipients of 100 to 300 × 106 cells had lesions, whereas 52% (32/61) of recipients of 300 to 600 × 106 cells were positive. Multiple transfers of sensitized cells were more effective than a single administration, with 33% to 86% of recipient animals showing thyroiditis. The transfer of thyroiditis required viable cells.
Radionuclide Marking of Canine Fetuses for Studying Ontogeny of the Immune Response
Authors: Stone J. M. , Shifrine Moshe
Published: December 2022

In all of the recent studies (1) on ontogeny of the immune response in mammals, a single antigen has been used to elicit the response in fetuses of one dam. Since individual fetuses in a litter can not be identified with this procedure, the following constraints are imposed: all fetuses must be injected, and with the same antigen; if different antigens are used a second injection can not be given; all sera have to be examined for antibody titers against all antigens used for injection; antibody responses can be compared only between litters, instead of within litters; different doses of an antigen can not be given to littermates; more animals need to be used for any given number of antigens.

Gamma-emitting radionuclides were selected for marking canine fetuses in utero because of their reasonably long physical half-lives (30 to 300 days), easily detectable energy emission spectra, low toxicity to biologic systems, commercial availability, and reasonable price.
Attempts to Sensitize Mice for PCA Using Pepsin Digested Mouse Antibody Fragments
Authors: Casey Francis B. , Tokuda Sei
Published: December 2022

Treatment of γG immunoglobulin with pepsin removes a portion of the molecule analogous to the Fc fragment leaving the F(ab′)2 fragment which has a sedimentation constant of approximately 5S (1). The biologic activity associated with F(ab′)2 fragments appears to vary with the species source of the antibody fragments and the test systems employed. For example, rabbit 5S F(ab′)2 antibody fragments, although incapable of sensitizing guinea pig skin for passive cutaneous anaphylaxis (PCA) (2), are capable of provoking PCA in the mouse (3). Additionally, F(ab′)2 obtained from rabbits and guinea pigs suppresses the development of the primary immune response in rabbits (4) and guinea pigs (5), respectively, whereas mouse F(ab′)2 is incapable of suppressing the primary immune response in mice (6). The purpose of this report is to describe experiments which demonstrate that while mouse F(ab′)2 preparations do not provoke PCA in the mouse, they specifically inhibit PCA induced by intact mouse antibody.
Consumption of Hamster Complement by Bacterial Endotoxin
Authors: Hook William A. , Snyderman Ralph , Mergenhagen Stephan E.
Published: December 2022

While studying the interaction of normal hamster serum incubated with endotoxin (LPS) to form a substance capable of releasing histamine from mast cells, it was observed that a concomitant loss of total hemolytic complement (C) activity occurred (1). Gewurz et al. have studied reaction mixtures containing guinea pig serum incubated with LPS and found that there was a preferential consumption of the terminal C components C3 through C9 (2). In contrast, we have found that LPS incubated in normal hamster serum under similar conditions caused a marked consumption of C1, C4 and C2 in addition to classical C EDTA (C3–C9). Accordingly, experiments were performed to compare C consumption by LPS with hamster and guinea pig serum under varying experimental conditions.

Blood was collected from female golden hamsters 5 to 10 weeks of age obtained from the Animal Production Unit, National Institutes of Health.
Antigen Quantitation in Experimental Immune Complex Glomerulonephritis
Authors: Wilson Curtis B. , Dixon Frank J.
Published: December 2022

Acute serum sickness glomerulonephritis was induced in rabbits with radiolabeled bovine serum albumin (I*BSA) 250 mg/kg. The kidneys of these rabbits contained 1 to 35 µg of I*BSA following >;99% elimination of I*BSA from the circulation. The I*BSA disappeared from the kidneys with a half time of about 10 days. The severity of glomerulonephritis in these rabbits was related to the rapidity of of immune elimination and the amount of I*BSA deposited. As little as 0.25 µg/g of kidney of I*BSA could be detected by immunofluorescence. During the 8-day period following immune elimination the BSA detectable by immunofluorescence decreased rapidly while host immunoglobulin and complement increased. The continued presence of I*BSA in these kidneys shown by isotopic studies suggested that this BSA was being covered by host anti-BSA antibody and complement from the circulation. These immunofluorescence findings occurred during the time of resolution of the glomerulonephritis by histologic and functional criteria. This suggested that the immune complexes formed initially in antigen excess were nephritogenic while immune complexes deposited in an extravascular site were less phlogogenic in spite of continued interaction with antibody and complement.
Inhibition of Target Cell Proliferation by Supernatants from Cultures of Human Peripheral Lymphocytes
Authors: Green J. A. , Cooperband S. R. , Rutstein J. A. , Kibrick S.
Published: December 2022

Supernatants from cultures of phytohemagglutinin (PHA)-stimulated human lymphocytes inhibit the division of certain cells in tissue culture. This effect of lymphocyte culture supernatant was routinely estimated by the rate of tritiated thymidine incorporation into DNA. In this system dilutions of 1:20 of the proliferation inhibitory factor (PIF) reduced DNA synthesis to 10% to 20% of that in control cultures. At dilutions of less than 1:640, PIF generally inhibited DNA synthesis by greater than 50%. Consistent with the decrease in DNA synthesis, cell cultures growing in the presence of PIF had fewer mitotic figures and lower total cell density than did control cultures. Supernatants from cultures established with dead leucocytes and PHA, extracts of fresh leucocytes, or PHA alone failed to produce this effect. A lesser degree of inhibitory activity also was present in supernatants from “unstimulated” lymphocyte cultures. Inhibition of proliferation in susceptible cultures was not accompanied by loss of viability; and cells from treated cultures excluded trypan blue, retained the ability to re-attach to glass, were capable of division and formed microcolonies when cloned. In addition to HeLa cells, susceptible cell cultures included human amnion, HEp 2, and KB cells. PIF was not effective in cultures of mouse L cells or in cultures of chick embryo fibroblasts. Preliminary data suggest that PIF is heat stable at 85°C for one-half hr, nondialyzable, nonsedimentable at 90,000 × G and is destroyed by trypsin.
Interrelationships of Tolerance and Immunity
Authors: Kong Yi-Chi M. , Wiger Donna L.
Published: December 2022

Passage of Armour bovine γ globulin (A-BGG) through the circulation of donor mice for 24 hr resulted in a preparation, P-BGG, which was essentially nonimmunogenic in recipient mice at a dose of 0.3 mg except when given in adjuvant. Immunoelectrophoresis of P-BGG against hyperimmune antiserum to A-BGG showed the presence of the major γ component but a lack of the minor fast γ component. P-BGG induced tolerance readily to the major component in adult mice; all mice challenged with P-BGG in Freund's adjuvant showed nonimmune elimination of labeled BGG and most showed either no hemagglutinins or 2-mercaptoethanol (ME)-sensitive antibodies with titers of 1:40 or below. When A-BGG was used for challenge, the hemagglutinins formed were primarily against the contaminant and were mainly ME-sensitive. In contrast, control mice given A-BGG in adjuvant and challenged with either antigen form showed immune elimination and ME-resistant antibodies with titers up to 1:2560. Attempts to prevent the removal of the phagocytizable components in A-BGG by blockade of the reticuloendothelial system with a suppressive dose of Thorotrast were unsuccessful. Pretreatment of donor mice with Thorotrast also did not affect the capacity of P-BGG to induce tolerance in the recipients.
Synthesis of the Ninth Component of Complement by a Clonal Strain of Rat Hepatoma Cells
Authors: Rommel Fred A. , Goldlust Marvin B. , Bancroft Frank C. , Mayer Manfred M. , Tashjian Armen H.
Published: December 2022

The MH1C1 strain of rat hepatoma cells was found to produce the ninth component of complement (C9). Most of the C9 produced by the cells was secreted into the culture medium. The production of C9 by MH1C1 cells was temperature sensitive with a Q10 of 2.2. Cycloheximide completely inhibited the production of C9 within 6 hr. Removal of cycloheximide at 24 hr resulted in a rapid return of C9 production. The production of C9 was also inhibited by treatment of the cells with actinomycin D. Hydrocortisone (3 × 10-6 M), which stimulates the production of serum albumin by MH1C1 cells 4- to 5-fold, did not increase C9 production. C9 produced by MH1C1 cells in culture was similar to guinea pig and rat serum C9 when comparisons were made by density gradient ultracentrifugation, by inhibition of hemolytic activity by antrypol, and by kinetic analysis of the reaction between C9 and EAC14235678. The results of these studies show that MH1C1 hepatoma cells synthesize C9 and secrete this complement component into the culture medium by processes that require both protein and RNA synthesis.
Heterogeneity of Immune Response to the Somatic (O) Antigens of Proteus Mirabilis
Authors: Smith James W. , Barnett Jack A. , Sanford Jay P.
Published: December 2022

A variation in the antibody response to antigenically differing somatic antigens (O) Proteus mirabilis was noted in rats. Certain strains induced the synthesis of 2-mercaptoethanol sensitive (2-MES), or IgM, antibody, whereas other strains led to the rapid appearance of 2-mercaptoethanol resistant (2-MER), or IgG, antibody. The varying response was not due to the differences in the quantity of lipopolysaccharide administered and was demonstrated when animals were challenged either separately or simultaneously with two strains. These differences in antibody response were not solely dependent upon antigenic mass administered as demonstrated by the failure of a 100-fold increase in antigenic mass to convert a 2-MES response to that of 2-MER, although a 100-fold diminution in antigenic mass converted the 2-MER response to 2-MES. No difference in the carbohydrate composition of the lipopolysaccharides obtained from these strains was apparent from gas chromatographic analysis. Thus, these studies demonstrate that the immune response of a given host can be determined not only by the quantity of antigen but also by differing characteristics of lipopolysaccharides from a single bacterial species.