bluuebunny/crossref_metadata_2025 huggingface dataset:
Studies on the Secretory Immunologic System of Fowl
Authors: Leslie Gerrie A. , Wilson H. R. , Clem L. William
Published: December 2022

The presence of immunoglobulins was investigated in the secretions of chickens. IgY was present in seminal plasma, tracheobronchial washes, alimentary tract washes and feces. In no case was it possible to demonstrate the presence of more antigenic determinants on these molecules than those present on serum IgY. Seminal plasma contained a 10.8S component that may represent a unique secretory molecule. IgM was not found in appreciable quantity in any of the secretions although trace amounts were detected in seminal plasma, feces, crop washes and duodenal washes. Intravenous administration of 125I-labeled IgY resulted in transfer of trichloracetic acid-precipitable radioactivity into seminal plasma, upper alimentary tract and feces. On the basis of these and other data it was concluded that the chicken has a secretory immunologic system wherein the secretory immunoglobulin is of the same class as the predominant serum immunoglobulin.
Partial Dissociation of Rheumatoid Synovial Fluid Cryoprotein: Micro-Complement Fixation by IgG- or IgG and IgM-Containing Fractions and Denatured Calf Thymus Dan
Authors: Marcus Robert L. , Townes Alexander S.
Published: December 2022

Complement-fixing cryoproteins from rheumatoid synovial fluid were partially dissociated using preparative ultracentrifugation in a 15% sodium chloride, sorbitol gradient. Heavy-sedimenting fractions retained capacity to fix human complement and contained variable amounts of DNA. Using a modified micro-complement fixation procedure, additional complement fixation was obtained upon addition of denatured calf thymus DNA to fractions containing IgG or mixed IgG and IgM. There was little fixation with native DNA. Interaction of heat-aggregated human γG globulin with denatured DNA under assay conditions failed to give micro-complement fixation in excess of that by γG globulin alone, reducing the likelihood that a nonspecific reaction occurs between denatured DNA and γG globulin aggregates. These findings demonstrated the presence in rheumatoid synovial fluid cryoproteins of immunoglobulin with determinants specific for a site or sites on denatured DNA.
Induction of Antigenic Competition with Thymus-Dependent Antigens: Effect on DNA Synthesis in Spleen Cells
Authors: Möller Göran
Published: December 2022

Thymus-dependent heterologous red cell antigens induced antigenic competition more efficiently than did thymus-independent lipolysaccharide antigens from Escherichia coli. The bacterial antigens responded to antigenic competition induced by other antigens to the same extent as red cell antigens. Mice subjected to antigenic competition exhibited a decreased level of DNA synthesis in spleen lymphocytes in comparison with normal mice.
Immunochemical Studies on Blood Groups
Authors: Lundblad Arne , Kabat Elvin A.
Published: December 2022

The immunochemical potency of the blood group A and B specific urinary difucosyl pentasaccharides was compared with that of mono- and difucosyl type 2 reduced oligosaccharides of soluble blood group A and B substances. The urinary difucosyl oligosaccharides were as active as the corresponding difucosyl compounds from blood group substance but were less effective inhibitors than the reduced monofucosyl blood group oligosaccharides in most sera and lectin systems used. With one A anti-A system, however, the urinary A difucosyl pentasaccharide was more active. The difucosyl B oligosaccharides gave slight inhibition in a Leb anti-Leb system whereas the difucosyl A pentasaccharide from urine was inactive.
Immunochemical Studies on Blood Groups
Authors: Feizi Ten , Kabat Elvin A. , Vicari Giuseppe , Anderson Byron , Marsh W. Laurence
Published: December 2022

Quantitative precipitin studies with three anti-I sera show them to be distinctly different in specificity as revealed by their reactions with milk fractions, with Lea substances, with precursor and with cow blood group substances and with the various stages of periodate oxidation and Smith degradation of A, B and H substances. The various I specificities are concealed in interior structures of the blood group A, B, H, Lea and Leb substances and may be exposed by stepwise periodate oxidation and Smith degradation of A, B and H substances or by mild acid hydrolysis of B substance. Oligosaccharide inhibition assays showed one anti-I serum (Ma) to be inhibited best by a terminal nonreducing β-D-Gal-(1 → 4)-β-D-GNAc-(1 → 6)-structure present in a precursor blood group substance. From quantitative precipitin studies, the specificity of another anti-I serum was entirely different. Evidence suggesting that certain anti-I sera may not contain homogeneous populations of anti-I molecules is presented. The genetic implications of the findings are discussed. Anti-I eluted from OI erythrocytes behaved similarly to the anti-I in the original antiserum. I-anti-I specific precipitates dissolve readily at 37°C and reprecipitate rapidly on colling. The definition of the human I-anti-I system based on hemagglutination reactions in the cold may not have revealed its ture complexity especially with respect to its genetics.
Deficient IgA Antibody Responses to Arsanilic Acid Bovine Serum Albumin (BSA) in Neonatally Thymectomized Rabbits
Authors: Clough John D. , Mims Linda H. , Strober Warren
Published: December 2022

The anti-hapten antibody response to intravenously injected arsanilic acid-bovine serum albumin was studied in neonatally thymectomized, 8-week-old rabbits using the Cla fixation and transfer test. This test allows quantitation of antibody in individual immunoglobulin classes. The IgG antibody response of thymectomized rabbits was not significantly different from the response of intact rabbits and the IgM antibody response of thymectomized rabbits was only slightly diminished. In contrast, the IgA antibody response of thymectomized rabbits was absent or strikingly diminished. This depressed IgA antibody response was also observed when animals were stimulated with a dose of antigen 10-fold greater than the original dose. It is proposed that for at least certain antigens, serum IgA antibody responses are more dependent on the thymus-derived cell system than are antibody responses in other immunoglobulin classes.
The Direct Hemolytic Plaque Response to Hemocyanin in the Rabbit
Authors: Kotoulas Angeliki O. , Moroz Leonard A.
Published: December 2022

Antibody-forming cells can be detected in the spleen and lymph nodes of rabbits immunized with keyhole limpet hemocyanin (KLH) by a direct plaque assay incorporating sheep red cells, coated with KLH by a chromic chloride method, as the target cell layer. After a single intravenous injection of associated KLH, direct plaque responses were observed in the spleen, with a peak at day 3, whereas no significant response was detectable in peripheral lymph nodes. When KLH in complete Freund's adjuvant was injected into hind footpads, the plaque response in regional lymph nodes was greater than that observed in the spleen. Numerically, the direct plaque response of rabbits to KLH was of the same order of magnitude as the response to bovine serum albumin reported by others.
Cellular Events during Induction of Immunologic Unresponsiveness in Adult Mice
Authors: Chiller Jacques M. , Weigle William O.
Published: December 2022

A specific and completely unresponsive state in adult mice was induced at the cellular level with ultracentrifuged human γ globulin (HGG). The cellular kinetics of induction of unresponsiveness were quantitated using a cell transfer system and the hemolytic plaque assay to detect plaque-forming cells to HGG. The results demonstrated that although 100% of the cells were not unresponsive until the 4th or 5th day after injection of tolerogen, 75% of the cells were unresponsive 6 hr following tolerogen. The kinetic curve did not reflect sequential induction to determinants located on the Fc and Fab fragments of HGG. Furthermore, the process of inducing unresponsiveness to HGG did not demonstrate antibody-forming cells detectable by the hemolytic plaque assay in the spleens of tolerogen-treated mice.
Immunologic Assays for Identifying Single Components in Protein Mixtures after Isoelectric Focusing in Urea-Containing Acrylamide Gels
Authors: Good Anne H. , Ceverha Barry B.
Published: December 2022

Procedures are described for locating either a specific antigen or a specific antibody in a mixture of proteins after isoelectric focusing in 5% acrylamide gels containing 8 M urea. The gels are frozen, cut into segments, and extracted with buffer. Complement fixation, passive hemolysis, or passive hemagglutination can then be performed to locate the gel segments that contain the desired material without prior removal of urea or ampholytes. Location of the desired component in a mixture by immunologic assay agrees well with the location of stained bands of that purified component when focused alone.
The Evolution of the Immune Response
Authors: Chartrand S. L. , Litman G. W. , Lapointe N. , Good R. A. , Frommel D.
Published: December 2022

Immunglobulins of two different sizes and classes that had antibody activity were purified from immune turtle serum. The characteristics of the IgM molecules did not deviate from those of molecules of this immunoglobulin class in other Placoderm-derived species. The low molecular weight immunoglobulin differed from mammalian IgG in that it had a sedimentation coefficient of 5.7, a shortened heavy chain with a molecular weight of 38,000, and low carbohydrate content with almost undetectable levels of hexosamine. This immunoglobulin had agglutinin activity, bound homologous and heterologous complement, and was transferred to the turtle egg. On the basis of the structural and biologic properties of this immunoglobulin, it was hypothesized that the C-terminal constant region of the heavy chains present in mammalian IgG was lacking. The implications of the molecular requirements for expression of the biologic function assigned to the Fc fragment were discussed in relation to diversification in function of immunoglobulins during evolution.
Further Characterization of Purified Fractions of M Protein from A Strain of Group A, Type 12 Streptococcus
Authors: Vosti Kenneth L. , Johnson Rudolph H. , Dillon Michael F.
Published: December 2022

Two purified fractions of M protein of type 12 streptococci were further characterized. Only one of the fractions, “b,” continued to meet additional physical, chemical and immunologic requirements for homogeneity. The protein in this fraction had a molecular weight of 32,000, contained N-terminal alanine and C-terminal leucine, stimulated formation of bactericidal antibodies and did not contain the nontype-specific determinants found in the “a” fraction. The differences in molecular weights and antigenic complexity disclosed by comparisons of the proteins of the “a” and “b” fractions suggested certain structural relationships among the molecules of M protein of type 12 streptococci.
Experimental Respiratory Syncytial Virus Infection of Adults
Authors: Mills V John , Van Kirk John E. , Wright Peter F. , Chanock Robert M. , Fishburne Isaac E.
Published: December 2022

Volunteers were inoculated with a low (102.7 pfu), or 200-fold higher dose (105.0 pfu) of respiratory syncytial virus (RSV). The low dose infected 100% of the men but did not produce illness; the level of virus replication and the magnitude of the subsequent antibody response were inversely correlated with the level of nasal secretory neutralizing antibody prior to infection. The high dose infected only 53% of the men (9 of 17); five of the infected men became ill. Two of the men who excreted virus apparently resisted experimental inoculation and acquired infection secondarily by contact with infected men. The differences in rates of infection between the two studies were minimized if only those men who had extensive infection (virus excretion τ; 102.5 pfu or significant antibody responses) were considered. Resistance to extensive infection in both studies, and resistance to illness in the second study, appeared to be correlated with high levels of nasal wash antibody, but not with the level of serum antibody.
Comparison of Lymphocyte Transformation, Inhibition of Macrophage Migration and Skin Tests Using Dialyzable and Nondialyzable Tuberculin Fractions from Mycobacterium Bovis (BCG)
Authors: Chaparas Sotiros D. , Thor Daniel E. , Hedrick Sally R.
Published: December 2022

Nondialyzable and dialyzable fractions prepared from BCG culture filtrates were compared in in vitro and in vivo tests for delayed hypersensitivity. With the non-dialyzable fractions in vivo lymphocyte transformation did not correlate with in vitro inhibition of macrophage migration or skin reactivity. The degree of lymphocyte transformation was correlated with the protein content of the fractions. A carbohydrate-rich fraction did not induce lymphocyte transformation but did induce skin reactivity and inhibition of macrophage migration. Activity of the dialyzable fractions in each of the three tests was dependent on molecular size.
In Vitro Response of Human Leukocytes to Associated and Dissociated Hemocyanin
Authors: Green Alexander A. , Borella Luis
Published: December 2022

Associated (98S) and dissociated (18S) KLH were prepared by rate zonal centrifugation. Peripheral blood leukocytes from unimmunized individuals were cultured with associated or dissociated KLH and the incorporation of 3H-TdR was determined after 6 days of culture. Associated KLH stimulated 3H-TdR incorporation 9 to 93 times (mean 38) more than that observed in control cultures without KLH. In contrast the mean incorporation of cultures incubated with dissociated KLH was only 1.3 times that in the controls. Leukocytes obtained from an immunized donor also responded to associated but not to dissociated KLH. We conclude that stimulation of DNA synthesis in human lymphocytes following in vitro exposure to KLH does not require prior in vivo exposure to this antigen, and that this stimulation depends on the molecular size of KLH.
An In Vitro Immune Response to Penicillin
Authors: Naor David , Henry Claudia , Fudenberg H. Hugh
Published: December 2022

The use of the in vitro system of Mishell and Dutton (1), originally described for the immune response of dissociated mouse spleen cells to red cells, has contributed valuable information to our understanding of the primary immune response. However, since the red cell is a complex and possibly atypical antigen, there are obvious advantages to extending the system with chemically defined haptens as antigens. Kettman and Dutton (2) have demonstrated a primary in vitro response to the determinant trinitrophenyl (TNP) using TNP-coupled sheep red cells as immunogen, and have also shown that the response is greatly enhanced when the spleen cells were taken from mice which had been immunized with the carrier, sheep red cells (3). Trowbridge et al. (4) obtained similar results using the hapten, hydroxy-nitroiodophenyl (NIP). Segal et al. (5) observed a primary in vitro response to the dinitrophenyl group (DNP) in spleen and lymph node explants.
Hapten-Carrier Relationships in the Production of Rat Homocytotropic Antibodies
Authors: Strejan Gill H. , Marsh David G.
Published: December 2022

Various hapten-protein conjugates have been used to elicit the production of antibodies involved in immediate-type hypersensitivity reactions in immediate-type hypersensitivity reactions in guinea pigs (1, 2), mice (3), rats (4, 5) and man (6). In the guinea pig, the concentration, the relative proportions of γ1 and γ2 and the average intrinsic association constant of anti-2,4-dinitrophenyl (DNP) antibodies were shown to depend on the nature of the carrier molecule used for immunization (7). Soluble extracts derived from Ascaris suum injected in association with B. pertussis vaccine elicited high titers of homocytotropic (Hc) antibodies in the rat; compared with hemocyanin and ovalbumin, 10 times less Ascaris antigen was required to induce detectable levels of Hc antibodies (8). The present report deals with the production of rat Hc antibodies to three different dinitrophenyl-carrier conjugates. The results will show that both anti-DNP and anti-carrier antibody titers obtained in rats immunized with a DNP-Ascaris conjugate were significantly higher than those obtained against DNP-hemocyanin or against DNP-bovine gamma globulin.
Human Clr, Purification and Assay Based on Its Linking Role
Authors: de Bracco M. , Stroud R. M.
Published: December 2022

C1, the first component of C, has been shown to be a macromolecule composed of three subunits, C1q, C1r, and C1s. Whereas C1q and C1s have been purified and monospecific antibodies have been produced against both, the C1r preparations used in most studies were only partially purified.

We developed a method of purification and assay for C1r. The purification procedure starts with a neutral euglobulin precipitation, followed by DEAE- and CM-cellulose chromatography, and a final preparative polyacrylamide gel electrophoresis.

Rabbits injected with this preparation produce monospecific precipitating antisera, which are capable of inhibiting C1 activity of cell-bound and fluid phase C1, but do not inhibit C1q or C1s.

The electrophoretic mobility of purified C1r is that of a β globulin on disc acrylamide electrophoresis at pH 8.6; its molecular weight, as estimated by Sephadex G-200 gel filtration, is 168,000.

The hemolytic assay of C1r in purified preparations is based on its property to link C1s to C1q and thereby generate macromolecular C1.
Restriction of the Substrate Specificity of C1 Human by Heat Inactivation or Cell Binding
Authors: Kondo M. , Gigli I. , Austen K. F.
Published: December 2022

Fluid phase or cell-bound activated first component of human complement (C1̄) utilizes the fourth (C4) and to a lesser degree the second (C2) components, as well as N-α-acetylglycyl-l-lysine methylester (AGLMe), N-acetyl-l-arginine methylester (AAMe), p-tosyl-l-arginine methylester (TAMe) and N-acetyl-l-tyrosine methylester (ATMe). The hydrolysis of either natural or synthetic substrates is decreased when C1̄ is cell bound (EAC1̄). Inactivation of C4 and C2 by C1̄ or EAC1̄ is inhibited by TAMe and AAMe, but not ATMe. C2 destruction by C1̄ in the presence of C4 exhibits a similar inhibition profile with these esters.

Heat-inactivated C1̄ loses the ability to split C2, TAMe, or ATMe, but not C4 or AAMe. C4 destruction by heat-inactivated C1̄ is prevented by AAMe. C1̄ can be blocked by C1̄INH so as to lose its activity against C2 and TAMe, but not C4 or AAMe.
Degradation of C3b by the C3b Inactivator
Authors: Ruddy S. , Austen K. F.
Published: December 2022

The C3b Inactivator (C3bINA) is a normal serum β globulin which destroys the activity of cell-bound C3b in immune hemolysis, immune adherence and the enhancement of phagocytosis. C3bINA treatment of EAC14oxy23 prepared with 125I-labeled C3 releases protein-bound radioactivity into the supernatant fluid. Kinetic studies demonstrate that the rate of disappearance of the hemolytic activity of cell-bound C3b greatly exceeds the rate of release of radioactivity from the cell surface. Polyacrylamide disc electrophoresis of the products of the action of C3bINA on washed EAC1423 125I cells reveals two new radioactive bands derived from the labeled C3, in addition to traces of C3b; only traces of C3b are found in supernatants from cells incubated in buffer alone. The more prominent new band migrates anodally to C3b, in a region identical to that of C3c produced by aging radioactive C3 in whole human serum at 37°C for 48 hr; the behavior of this band on gel filtration also resembles that of C3c and this band contains the “A” antigen of C3.
C3 Consumption in Immune Reactions
Authors: Pondman K. W. , Hannema A. , Wolters G.
Published: December 2022

C3 consumption and site formation was studied with specific antisera against three antigenic determinants of the C3 globulin molecule, namely, [β1C] or B, the β1A and the α2D determinant.

Interaction between C3 in serum and optimal specific precipitates results in the rapid loss of the [β1C] or B determinant and concomitant appearance of C3i and free α2D in the fluid phase, the latter determinant being nearly unexposed in intact C3 globulin. Experimental data further suggest that the B determinant of the intact C3 molecule is lost by destruction or by intramolecular rearrangement in the C3 molecule after interaction with cell bound C. This is borne out by the following data. (a) Immune aggregates with bound complement are incapable of inducing anti-[β1C] antibodies in rabbits; (b) sensitized Lea-positive red cells with bound complement agglutinate with monospecific anti-α2D and anti-β1A, but fail to agglutinate with anti-[β1C]; (c) EAC1̄, cells prepared with limited C3 fail to agglutinate with anti-[β1C] but agglutinate strongly with anti-β1A and anti-α2D.
Additional Studies on the Fractionation of Human C5: Identification, Possible Method of Removal and Significance of a Plasminogen/Plasmin Contaminant
Authors: Nilsson U. R. , Tomar R. H. , Taylor F. B.
Published: December 2022

Preparations of C5 purified according to a previously described preparation procedure (Nilsson and Müller-Eberhard, 1965) demonstrated fibrinolytic activity when reacted with streptokinase in a bovine clot lysis assay. Fibrinolytic activity was readily demonstrable although quantitative analysis revealed that the degree of contamination would correspond to only 0.01% to 0.05% plasminogen/plasmin (Plg/Pln). Presence of a potentially active proteolytic enzyme in the C5 fractions may cause uncontrolled modification of the C5 protein. Loss of C5 hemolytic activity upon treatment with relatively large doses of Pln supports this possibility. Attempts have been made to accomplish a complete separation of C5 from Plg/Pln, as well as to prevent possible proteolytic action by Pln during the purification procedure.
A Blood Coagulation Abnormality in C6 Deficient Rabbits and Its Correction by Purified C6
Authors: Zimmerman T. S. , Arroyave C. M. , Müller-Eberhard H. J.
Published: December 2022

In this communication we report a coagulation abnormality in rabbits genetically deficient in the sixth component of complement which is characterized by prolonged whole blood clotting time and deficient prothrombin consumption. The mean glass clotting time in five C6 deficient rabbits was 38 min (range: 34 to 41 min) as opposed to a mean of 24 min (range: 20 to 31 min) in five normal rabbits. Clotting times in plastic tubes varied from 140 to over 400 min in C6 deficient animals, whereas the range for the normal rabbits was 70 to 90 min. Prothrombin consumption measured in plastic tubes for 1 hr at 37°C varied from 0% to 10% in six deficient rabbits, whereas the mean for six normal rabbits was 72.5% with a range of 62% to 79%. Prothrombin and partial thromboplastin times were normal in the deficient animals as were activities of the classical clotting factors II, V, VII, VIII, IX, X, XI and XII.
Arrangement of Macromolecular Components in Erythrocyte Membranes and Their Relation to Bound C3 and C4
Authors: Dalmasso A. P. , Diaz A. , de Bracco M. M. E.
Published: December 2022

The purpose of this study was to examine the distribution of a ntigenic components in the intact erythrocyte (E) membrane (M) and their relation to bound C3 and C4. Rabbit antisera were prepared by immunization with whole sheep EM. Immunoelectrophoresis (IEP) of M solubilized with 2.5% Triton X-100 (T) revealed one β2 line and two to four α-β1 lines. M solubilized by sonication in 10% n-butanol and fractionated by ultracentrifugation in 24% sucrose (Morgan, T. H. and Hanahan, D. J., Biochemistry, 5: 1050, 1966) yielded a light fraction rich in carbohydrates and lipids, having high affinity for T and β2 position on IEP after treatment with T, and a heavy fraction rich in protein, migrating in the α-β1 IEP area. Absorption of anti-EM with intact E removed its hemagglutinating activity and its capacity to react with the β2 component without altering its anti α-β1 activity.
Complement-Mediated Lysis of Nucleated Cells
Authors: Shipley U. W. , Baker A. R. , Colten H. R.
Published: December 2022

The availability of a rapidly growing cell line in tissue culture (Chinese hamster lung, CHL) with high plating efficiency and the ability to form discrete colonies has made it possible to adapt the colony inhibition technique to quantitative studies of complement-mediated lysis of nucleated cells and, in particular, to study the effect of the cell growth cycle on the susceptibility of target cells to immune lysis. During complement-mediated lysis, the cells were examined by phase contrast microscopy, the molecular weight of target cell DNA was estimated by ultracentrifugation in alkaline sucrose gradients and the extent of lysis was determined by a loss of colony-forming ability.

Within 8 min after the addition of complement to sensitized CHL cells, morphologic changes in the target cells were noted. Following these changes, an enzymatic degradation of target cell DNA occurred, resulting in a 10-fold reduction in molecular weight of the single-stranded DNA fragments.
The Immune Response in the Hamster
Authors: Coe J. , Peel L. , Smith R. F.
Published: December 2022

Hamster 7Sγ1 and 7Sγ2 antibodies to hen-egg albumin (HEA) were tested for their ability to fix complement and mediate passive cutaneous anaphylaxis (PCA). Only 7Sγ2 anti-HEA bound complement in the micro-complement-fixation test and hemolysed HEA-sensitized sheep red blood cells. In addition, only 7Sγ2 anti-HEA mediated the PCA reaction in guinea pigs, which could be blocked with 7Sγ2 globulin from normal hamster sera. Hamster antibody did not produce detectable PCA reactions in hamsters, and hamsters with circulating 7Sγ1 anti-HEA did not show signs of active anaphylaxis after intravenous challenge with the antigen.
Genetic Polymorphism in Rhesus C3 and Ge Globulin
Authors: Alper Chester A. , Robin Noel I. , Refetoff Samuel
Published: December 2022

Extensive genetic polymorphism of C3 was found in 81 sera from rhesus monkeys. Evidence was obtained for three alleles, rhesus C3S, C3F and C3F1, with gene frequencies of 0.66, 0.33 and 0.01. The variants were inherited as autosomal codominant traits. Polymorphism of Gc globulin was also found. Although most sera contained Gc globulin that formed a single band on electrophoresis, in a few sera this protein consisted of two bands, one of which was slower than the usual. These observations suggested that Gc globulin was controlled by two codominant alleles, rhesus Gc1 and Gc2, with frequencies of 0.975 and 0.025, respectively.
Antibody Formation Against Salmonella Flagella by an Immune Ribonucleic Acid Fraction
Authors: Yamaguchi Nobuo , Kurashige Satonori , Mitsuhashi Susumu
Published: December 2022

An immune ribonucleic acid (RNA) preparation was extracted with phenol from the spleens of mice immunized with Salmonella flagella. A high titer of serum antibody was produced after injection of only a small amount of Salmonella flagella into mice that had previously been injected with immune RNA. This reaction was found to be immunologically specific and indicated a secondary antibody response. In contrast, secondary responses were not induced by injection of an immune RNA preparation into mice primed with either an immune RNA preparation or Salmonella flagella. These results suggest that immune RNA preparations did not contain antigens or fragments thereof, or at least the antigens that are needed to stimulate a secondary response. They indicated, moreover, that the immune RNA preparation can induce memory cells capable of responding to a secondary antigenic stimulus. The required interval between first and second antigenic stimulation and the necessary amounts of immune RNA or flagella antigen were investigated.
Effect of Cold Exposure on the Immunologic Response of Rabbits to Human Serum Albumin
Authors: St. Rose J. E. M. , Sabiston B. H.
Published: December 2022

The effect of low environmental temperature on the response of rabbits to a single injection of human albumin was examined. Animals exposed to cold (4°C ± 2°C) had faster catabolism of antigen and earlier appearance and higher levels of circulating antibody. Mercaptoethanol-resistant antibody was produced early (12 days) in coldexposed animals and late (26 days) in non-exposed animals. The effects of cold exposure on the immune response were as apparent after acute exposure (1 to 2 days) as after chronic exposure (3 to 8 wk).
Inability of the NZB/NZW F1 Thymus to Transfer Cyclophos-Phamide-Induced Tolerance to Sheep Erythrocytes
Authors: Jacobs Michael E. , Gordon Joyce Kay , Talal Norman
Published: December 2022

NZB/NZW (B/W) and C57BL/6 mice were made tolerant to sheep erythrocytes with cyclophosphamide. Thymus or bone marrow cells from tolerant or untreated donors were injected in various combinations into syngeneic irradiated recipients and assayed for plaque-forming cells. In C57BL/6 mice both marrow and thymus transfer tolerance; however, in B/W mice only the bone marrow did so. Unexpectedly, the B/W thymus from tolerant donors cooperated fully with B/W marrow cells from untreated donors. Thus the B/W thymus did not express the donor animal's tolerance status.
Immunochemical Studies on Bovine Carboxypeptidases a
Authors: Amiraian Kenneth , Plummer Thomas H.
Published: December 2022

Rabbit antisera to bovine carboxypeptidase Aγ were prepared by intravenous injection of a crystalline suspension of the enzyme. Quantitative immunoprecipitation tests with these antisera and carboxypeptidases Aγ and Aα and procarboxypeptidase A show that carboxypeptidase Aα, which possesses seven additional amino acids in the amino terminal position compared with carboxypeptidase Aγ, precipitates 10% less antibody N than does the homologous antigen. Procarboxypeptidase A precipitates only 8% to 15% of the antibody N as carboxypeptidase Aγ, and the proenzyme is maximally precipitated at the same weight of N rather than at the same molar concentration as the enzyme. Immune precipitates formed with carboxypeptidase Aγ and rabbit anticarboxypeptidase Aγ sera, at equivalence, are almost as active as the free enzyme in hydrolyzing carbobenzoxyglycyl-L-phenylalanine. These precipitates are capable, to a lesser degree, of hydrolyzing the carboxy terminal amino acids of human and bovine serum albumins. Immune precipitates formed with bovine carboxypeptidase B and its rabbit antisera, at equivalence, are as active as the free enzyme toward the substrate hippuryl-L-arginine.
Immunochemistry of the Cell-Wall Carbohydrate of Group L Hemolytic Streptococci
Authors: Karakawa W. W. , Wagner J. E. , Pazur J. H.
Published: December 2022

A cell-wall carbohydrate consisting of N-acetylglucosamine, N-acetylgalactosamine, galactose and rhamnose was isolated from group L hemolytic streptococci. The antigenic determinant of this carbohydrate consisted of terminal β-N-acetylglucosamine residues attached to rhamnose side chains of the polymer. Group L carbohydrate treated with an induced enzyme preparation from a soil bacterium no longer gave a precipitin reaction with its homologous antiserum but did so strongly with antigroup G streptococcal serum. The native carbohydrate also gave a precipitin reaction with antisera from rabbits immunized with group A streptococci. The antigenic cross-reactivity between group A and group L carbohydrates was due to the fact that the antigenic determinants of both carbohydrates were terminal β-N-acetylglucosamine residues. The N-acetylglucosamine residues of group L carbohydrate were extremely acid labile, whereas those of the group A carbohydrate were considerably more stable. This difference indicated some subtle structure differences between group A and group L antigens.
Distribution of H-2 Alloantigenic Specificities on Radiolabeled Papain-Solubilized Antigen Fragments
Authors: Cullen Susan E. , Nathenson Stanley G.
Published: December 2022

A method for determining the distribution of H-2 alloantigenic specificities on papain-solubilized fragments has been developed. Partially purified, radiolabeled antigen was reacted with monospecific alloantiserum and the antigen-antibody complex was removed by gel filtration or disc gel electrophoresis. Several specificities were shown to reside on the same fragment because monospecific sera removed specificities in addition to the one against which each serum was directed.
In Vitro Studies of the Human Response to Organ Allografts: Appearance and Detection of Circulating Activation Lymphocytes
Authors: Hersh Evan M. , Butler William T. , Rossen Roger D. , Morgan Robert O. , Suki Wadi
Published: December 2022

The rate of 3H-thymidine incorporation into circulating peripheral blood lymphocytes was measured immediately after isolation from the blood of 24 organ allograft recipients followed serially, and 42 normal control subjects. The isotope incorporation by 106 cells incubated for 2 hr with 2 µCi of 3H-thymidine was referred to as the activated lymphocyte level. It was 319 ± 295 (mean ± standard deviation) counts per minute (cpm) in the normal controls, 535 ± 377 cpm in the patients prior to grafting, 644 ± 629 cpm after grafting but when the patients had adequate nonfluctuating graft function, and 6744 ± 3798 cpm during active rejection characterized by progressively decreasing graft function. Levels rose an average of 5.4 days prior to clinical signs of rejection and tended to remain high for an average of 10.9 days after rejection was controlled, as characterized by stabilization or improvement in graft function. This outpouring of dividing lymphoblasts was interpreted as the production of effector cells in response to the antigenic stimulus of the graft. Its identification may be useful to predict impending rejection.
Regulation of Homocytotropic Antibody Formation in the Rat
Authors: Taniguchi Masaru , Tada Tomio
Published: December 2022

The effects of four representative immunosuppressants, i.e., 5-bromouridine deoxyriboside (BUDR), actinomycin D, cyclophosphamide and cortisone, on the formation of homocytotropic antibody (HTA) were observed in rats immunized with dinitrophenylated ascaris extracts (DNP-As) and killed Bordetella pertussis, with the following results. 1) A single injection of 35 mg/kg of BUDR given simultaneously with or shortly after the start of immunization caused enhancement and prolongation, after some delay, of HTA formation, as measured by the mean of reciprocal PCA titers. 2) Pretreatment with 0.3 mg/kg of actinomycin D caused a delayed response of HTA, but the subsequent mean peak titer was about the same as that of the control untreated rats. Such treatment after immunization was started caused an absolute stimulation of HTA, as judged by high peak titers and prolonged production of the antibody. 3) Pretreatment with a single injection of 150 mg/kg of cyclophosphamide caused delayed but enhanced formation of HTA which lastedo lnger than the normal period of time, whereas the same treatment started 2 days after the antigen was given caused suppression. 4) Pretreatment with 200 mg/kg of cortisone caused temporary enhancement, whereas treatment during immunization was moderately suppressive. The enhancement or suppression of HTA formation did not correlate with γM and γG antibodies in the serum, as measured by passive hemagglutination titers before and after 2-mercaptoethanol treatment.
Long-Term Reconstitution of Bursectomized Chickens
Authors: Cain W. A. , David L. A.
Published: December 2022

Since the discovery of the immunologic role of the avian bursa of Fabricius (1), several investigations (2–4) have succeeded in defining functions of this organ. Bursectomy before hatching (5, 6), treatment of embryos or young chicks with testosterone (7) or cyclophosphamide (8), and bursectomy on the day of hatching coupled with whole body x-irradiation (9) block development of bursa-dependent components of peripheral lymphoid tissues.

In later life, chickens bursectomized by one of these methods are deficient in immunoglobulins, lymphoid germinal centers, plasma cells and the ability to make antibodies. It can be concluded that the bursa of Fabricius is the central lymphoid organ in chickens, which performs crucial steps in the development of humoral immunologic competence.

Less progress has been realized in attempts to define mechanisms of bursal action.
A Simplified Method for Measuring Human IgE
Authors: Centifanto Ysolina M. , Kaufman Herbert E.
Published: December 2022

The radioactive single radial diffusion test described by Rowe (1) is commonly used for determining human IgE in secretions. Although this test is sensitive and allows the measurement of IgE at amounts as low as 40 ng/ml, it has definite drawbacks. First, extensive washing is necessary to remove both unreacted sheep antihuman IgE and 131I-labeled rabbit antisheep globulin serum. Second, the short half-life of the radioactive label makes this test impractical for routine diagnostic measurements of IgE in secretions or sera.

Another test described by Arbesman and Ito (2) uses a one-step reaction with 125I-labeled antihuman IgE serum, thus eliminating the need to use antibodies to anti-IgE.

Although both tests are sensitive and very suitable for research projects, a simplified procedure that does not require radioactive labeling seemed desirable. For this purpose, we developed a new method to detect IgE in human tears, which employs a fluorescein-labeled rabbit antisheep globulin reagent.
Characterization of Immunoglobulin G Anti-Glomerular Basement Membrane Antibodies Eluted from Kidneys of Patients with Glomerulonephritis
Authors: McPhaul John J. , Dixon Frank J.
Published: December 2022

Direct immunofluorescence microscopy was performed on sections of kidney from 22 patients with anti-glomerular basement membrane (anti-GBM) antibody-mediated nephritis proved by in vitro elution techniques. Although immunoglobulin G (IgG) was fixed to glomeruli of all native or transplanted kidneys, the third component of complement (C3) was bound at the site of IgG fixation in slightly more than half (10 of 17). Immunoglobulin subtyping of eluted IgG anti-GBM antibodies, performed by an indirect immunofluorescence technique, indicated the presence of IgG subtypes 1, 2 and 3 in the eluates. Although eluted IgG anti-GBM antibodies failed to fix human C3 on normal homologous kidney sections, IgG anti-GBM antibodies isolated from serum of nephritic patients did so. Failure to visualize C3 by immuno-fluorescence examination of nephritic human kidneys was usually not due to an anomaly of IgG subtype participation or to lack of intrinsic ability of IgG anti-GBM antibodies to bind C3 in vitro.
Anti-Neuraminidase Antibody Response in Serum and Nasal Secretions Following Intranasal or Subcutaneous Inactivated A2/Hong Kong/68 Influenza Virus Vaccine
Authors: Fedson David S. , Fulk Robert V. , Huber Margret A. , Reisberg Margaret A. , Kasel Julius A.
Published: December 2022

The anti-neuraminidase antibody (ANAb) response following subcutaneous or intranasal A2/Hong Kong/68 inactivated influenza virus vaccine was evaluated in 81 volunteers of varying ages. Rises in serum ANAb titer were seen in 33 subjects although 76 developed neutralizing antibody rises. In nasal secretions ANAb rises appeared in only 12 subjects as compared with 31 subjects with neutralizing antibody rises. In individual subjects the magnitude of serum ANAb rises to the Hong Kong neuraminidase and an earlier related A2/1957 neuraminidase were similar. Serum and secretory ANAb cross-reacted considerably with neuraminidase antigens of earlier strains of A2 influenza virus. The enzyme inhibition test for ANAb was more sensitive than the hemagglutination inhibition test. ANAb in nasal secretions was IgA. Since ANAb may participate in host defense against influenza virus infection, it may be important to measure the antibody response to the neuraminidase as well as hemagglutinin antigen to select vaccines with optimal prophylactic activity.
C3b Inactivator of Man
Authors: Ruddy Shaun , Austen K. Frank
Published: December 2022

C3b inactivator (C3bINA), a normal serum protein that destroys the ability of cells bearing C3b to participate in immune adherence, to undergo enhanced phagocytosis by polymorphonuclear leukocytes and to react with subsequent components of the complement system, was studied. C3bINA treatment of EAC14oxy23 cells prepared with purified and radiolabeled C3 released two bands of radioactive protein that were distinguishable from C3b on polyacrylamide disc gel electrophoresis. Similar products were demonstrable following the interaction of fluid phase C3b with C3bINA. The more anodal band appeared to be identical to C3c or β1A on the basis of electrophoretic mobility, behavior on gel filtration and antigenic content. The portion of C3 that remained on the cell following C3bINA treatment resembled C3d or α2D in its antigenicity. The action of C3bINA in vivo to produce cells coated only with C3d could account for absence of immune adherence reactivity and resistance to lysis of erythrocytes circulating in patients with certain forms of hemolytic anemia.
Immunochemical Assay of Antilymphocyte Serum with Subcellular Fractions and Purified Membranes
Authors: Von Fellenberg Roland , Briner Hans U. , Guggisberg Elisabeth
Published: December 2022

Highly immunosuppressive and non-suppressive antilymphocyte sera (ALS) were compared by quantitative micro-complement fixation. Skin graft survival was correlated with micro-complement fixation properties using purified membranes of spleen cells as antigen. The immunochemical anti-membrane activity of the ALS was due to both specific antigens of lymphoid cells and non-cell-specific antigenic determinants. The results obtained immunochemically were confirmed morphologically by indirect immunofluorescence staining. The biologic activity of ALS was discussed with respect to cooperation between specific and non-cell-specific antibodies.
Relative Absence of Circulating Antigen-Reactive Cells during Oral Immunization
Authors: Goldberg Salmon S. , Kraft Sumner C. , Peterson Raymond D. A. , Rothberg Richard M.
Published: December 2022

Adult rabbits subcutaneously immunized with bovine serum albumin (BSA) incorporated in incomplete Freund's adjuvant regularly had circulating antigenreactive cells (ARC) 14 to 21 days later. During a 3-month study period, animals immunized by oral administration of BSA in low concentrations rarely had detectable ARC despite the presence of comparable amounts of circulating anti-BSA. Discontinuing antigen administration led to diminished antibody concentrations but a persistent absence of detectable circulating ARC. Following a subcutaneous injection with BSA in incomplete Freund's adjuvant that resulted in detectable circulating ARC in non-immunized rabbits, orally immunized animals developed an increased rate of antibody synthesis without detectable ARC. The ability of lymphocytes from orally immunized animals to respond to BSA was demonstrated in 64% of these rabbits 1 week after 100 mg BSA was given intravenously after 3 months of antigen ingestion. These results and studies by others suggested that during oral immunization, ARC primarily are localized to gut-associated lymphoid tissue.
The Nature of the Antibody in Iso-Immune Anti-L Sera Affecting Active Potassium Transport in LK Sheep Red Cells
Authors: Snyder J. J. , Rasmusen B. A. , Lauf P. K.
Published: December 2022

The factor in ovine iso-immune L-antiserum that was responsible for stimulation of active K+-transport in LK sheep red cells was found to be an antibody of the IgG1 class. Reduction and alkylation of the anti-L-IgG1 protein abolished its K+-pump stimulatory activity. Treatment with papain to yield univalent Fab fragments substantially reduced stimulatory activity; there was no enhancement from the Fc protein. Bivalent fragments obtained by pepsin digestion, however, were nearly fully active in effecting stimulation of active K+-transport in LK sheep red cells.
Isolation and Physico-Chemical Characterization of the “IgM-Like” Immunoglobulin from the Stingray Dasyatis Americana
Authors: Johnston William H. , Acton Ronald T. , Weinheimer Peter F. , Niedermeier William , Evans E. Edward , Shelton Emma , Bennett J. Claude
Published: December 2022

The stingray Dasyatis americana was found to synthesize high molecular weight antibodies against Salmonella typhosa “H” antigen and human A erythrocytes. These antibodies were demonstrated to be macroglobulins with a sedimentation coefficient of 17.1S and a molecular weight of 820,000. Carbohydrate analysis revealed a total content of 9.3% and a composition which was distinctively different from human IgM. Complete reduction and alkylation of the stingray macroglobulin, followed by gel filtration in the presence of guanidine hydrochloride, resulted in separation of the heavy (H) and light (L) polypeptide chains. These chains were found to have molecular weights of 70,000 and 23,000 for the H and L chains, respectively. With the exception of a lower content of the basic amino acids, the amino acid composition of the H and L chains was similar to that reported for human macroglobulin polypeptide chains. The stingray macroglobulin was examined by electron microscopy and displayed a pentameric structure. The data suggest a structural similarity between stingray macroglobulin and mammalian IgM.
Early and Enhanced Germinal Center Formation and Antibody Responses in Mice after Primary Stimulation with Antigen-Isologous Antibody Complexes as Compared with Antigen Alone
Authors: Laissue Jean , Cottier Hans , Hess Max W. , Stoner Richard D.
Published: December 2022

Primary antibody responses elicited in mice by injection into the hind leg footpads of tetanus toxoid complexed with isologous antibody at equivalence were enhanced as compared with responses to antigen alone. Serum antitoxin titers in animals given immune complexes started to rise approximately 4 days earlier and reached peak values more than 15 times higher than in mice given antigen alone. In both groups of animals, however, the doubling time of serum antitoxin concentrations was similar and approximated 30 hr as compared with 8 hr in anamnestic responses to tetanus toxoid. These differences were reflected in de novo appearance and growth of germinal centers in popliteal lymph nodes. The median number of germinal centers/lymph node cross-section of complex-injected animals began to rise significantly 4 days after stimulation and throughout days 4 to 16 exceeded corresponding values for mice given tetanus toxoid. The median numbers of germinal centers in both groups of stimulated animals was maximal around day 12 and fell to near-control values at day 20, indicating that many of these structures underwent involution within 3 wk following de novo formation. The potentiating effect of antigen complexes as compared with antigen was seen in the significantly larger size of newly formed germinal centers from day 8 through day 24 following stimulation. The best estimate for volume doubling time of developing germinal centers after injection of immune complexes was 10 hr or less, which was near corresponding values proposed previously for germinal center growth in regional lymph nodes during anamnestic responses. Labeling patterns 1 hr after i.v. injection of 3H-thymidine and mitotic activity of lymphoid germinal center cells revealed no significant differences between the two groups of stimulated mice. These results were discussed in view of available evidence that identified lymphoid germinal center cells as precursors of antibody-forming elements possibly related to production of a certain class or subclass of immunoglobulins. The question was raised whether in vivo formation of immune complexes is also instrumental in triggering the germinal center system in the course of a true primary antibody response to antigen alone.