bluuebunny/crossref_metadata_2025 huggingface dataset:
Inactivation of Bacteriophage T4, of Poly-D-Alanyl Bacteriophage and of Penicilloyl Bacteriophage by Immunospecifically Isolated IgM and IgG Antibodies
Authors: Haimovich Joseph , Sela Michael
Published: December 2022

Anti-bacteriophage T4, anti-poly-d-alanyl and anti-penicilloyl antibodies were isolated from the sera of goats and rabbits at different times after immunization by means of water-insoluble immunoadsorbents prepared with bromoacetyl cellulose. The isolated antibody preparations were separated into IgM and IgG fractions. The relative efficiency of the isolated IgM and IgG antibodies in inactivating bacteriophage T4, poly-d-alanyl bacteriophage and penicilloyl bacteriophage differed with respect to the time after injection at which sera were collected, the method of plating and the specificity of the respective antibody used for the inactivation, leading to the conclusion that it is not possible to determine the respective concentration of antibodies of the two classes exclusively from phage inactivation measurements.
The Immunologic Response of the Mouse to Aerosols of an Attenuated Encephalomyocarditis Virus Strain
Authors: Prato Catherine M. , Akers Thomas G.
Published: December 2022

The antibody response, interferon synthesis, viremia and replication of virus in the lungs of mice following aerogenic immunization with mengovirus-37A were studied. Virus titers in blood and lungs were maximal at day 1, whereas interferon activity was greatest at day 7 after aerosol exposure. Hemagglutination-inhibition (HAI) activity (2-ME sensitive) was detected on day 6, neutralizing antibodies by day 9 (also 2-ME sensitive) and complement fixation tests were positive on day 12. After day 12, 2-ME resistant HAI and neutralizing antibodies were observed. Maximum HAI and neutralizing antibody titers occurred on day 31.

The usual pattern of early 19 S response followed by 7 S antibody formation was observed following aerogenic immunization of mice with mengo virus-37A. However, the time required for antibody to appear was longer than that noted in other immunization studies using other routes of vaccination.
Identification and Genetic Control of Two New Low-Density Lipoprotein Allotypes: Phenogroups at the Lpq Locus
Authors: Albers John J. , Dray Sheldon
Published: December 2022

Two new rabbit low-density lipoprotein allotypes, Lpq-3 and Lpq-4, have been identified.

Segregation of the two allotypes is consistent with control by allelic genes.

The four allotypes at the Lpq locus, q1, q2, q3, and q4, are inherited as phenogroups controlled by three co-dominant alleles q1, 3, q2, 3, and q2, 4.
Comparative Immune Response to Amphibian Cytoplasmic Viruses Assayed by the Complement-Fixation and Gel Immunodiffusion Tests
Authors: Kaminski Suzanne , Clark H. Fred , Karzon David T.
Published: December 2022

Rabbits and guinea pigs immunized with several amphibian cytoplasmic virus strains consistently responded with neutralizing, complement-fixing, and precipitating virus-specific antibodies. Complement-fixation and gel-immunodiffusion tests revealed a close antigenic relationship among all strains tested, in agreement with previous results obtained with the virus-neutralization test. No antigenic relationship was demonstrated between the amphibian viruses and the morphologically similar Tipula iridescent virus.

Bullfrogs and snapping turtles responded irregularly to amphibian virus inoculation with low-titered neutralizing and/or precipitating antibody. However, antibody was detected in pre-immunization sera of some frogs and turtles, suggesting prior exposure to related viruses.

Viral antigen was not detected in leopard frog renal tumors by the immunodiffusion or complement-fixation tests. Similarly, antibody to cytoplasmic virus antigen could not be detected in sera of tumor-bearing or normal leopard frogs.
Studies on the Antigenicity of Bacterial Ferredoxins
Authors: Nitz Rodney M. , Mitchell Barbara , Gerwing Julia , Christensen John
Published: December 2022

The native ferredoxin of Clostridium pasteurianum and its performic acid-oxidized derivative were used as antigens to obtain antisera from rabbits. The antibodies produced were detectable by complement fixation and immune precipitation. Cross-reactivity studies were carried out using these antigens, ferredoxin from which the iron had been removed by trichloroacetic acid (TCA)-precipitation, and ferredoxin that had been reduced and alkylated with iodoacetamide. In all cases considerable degrees of cross-reactivity were observed. The results imply that the cysteinyl residues of C. pasteurianum ferredoxin may not be critically involved as antigenic determinants.
Studies on Lymphocyte Transformation in Nephrotoxic Serum Nephritis of the Rat
Authors: Warnatz Hermut , Scheiffarth Friedrich , Hagel Friedrich
Published: December 2022

Nephrotoxic serum nephritis was produced in 18 Wistar rats by injection of an heterologous anti-rat kidney serum. From 3 to 5 weeks after the first intravenous injection all animals showed the typical histologic lesions of nephritis. With the immunofluorescence technique the localization of heterologous globulin to the basal membranes of the glomeruli was demonstrated. In four cases a strong fixation of rat γ globulin was observed. Kidney autoantibodies were detected with the passive hemagglutination technique. Precipitating antibodies against kidney extract were absent in all cases.

Neither autologous kidney extract nor nephrotoxic rabbit serum and normal rabbit serum caused an increased transformation of lymphocytes obtained from rats with nephrotoxic serum nephritis. On the contrary, cultures of lymphocytes from these rats showed a depression of blast cell formation after being exposed to nephrotoxic serum in comparison with lymphocytes of normal rats. This depression was more intense in cultures of lymph node lymphocytes than in those of thoracic duct lymphocytes. The results indicate that lymphocytes do not play a role in the pathogenesis of nephrotoxic serum nephritis. The possible lymphocytotoxic activity of the nephrotoxic serum, however, should be taken into consideration for the judgement of these results.
High Efficiency of SAClgp4gp2gp Conversion to S* with EDTA-Treated Rat Serum
Authors: Miyakawa Yuzo , Sekine Teruaki , Shimada Kohkichi , Nishioka Kusuya
Published: December 2022

The conversion of SAC1gp4gp2gp to S* in titrations of guinea pig C1, C4 and C2 is much more efficient with Crat-EDTA than with Cgp-EDTA. Comparative titrations indicate that the efficiency of this conversion by the usual procedure with Cgp-EDTA is only 6% or less.
Enhancing Antibody
Authors: Zimmerman Barry , Feldman Joseph D.
Published: December 2022

While studying the enhancement of normal skin homografts in rats, we have observed that Lewis (Le)2 antisera created against either Buffalo2 (Bf) or Brown Norway (BN)3 rat tissues prolonged survival of test skin grafts taken from both donor strains. In each case, however, the antiserum prolonged the survival of its specific homograft to a greater degree than the cross-reacting graft. The data presented here indicate that Bf and BN rats share histocompatibility antigens, and that Le anti-Bf enhancing sera contain antibodies to more than one transplantation antigen.

The production of enhancing sera and their assay for prolongation of skin homograft survival on 10- to 12-day-old Lewis rats2 3 have been previously described (1). Briefly, 200- to 250-g male rats received first and second set skin grafts followed by two weekly infusions of 20 to 30 × 106 donor spleen cells. One week after the last infusion the serum was harvested.
The Latency Requirements and Rate of Evolution of Passive Cutaneous Anaphylactic Reactions Produced by Untreated and Carbamylated Rabbit γG Antibody
Authors: Feinman Lawrence , Cohen Stanley , Becker Elmer L.
Published: December 2022

The amounts of antibody necessary to produce passive cutaneous anaphylaxis (PCA) with and without a latent period were compared, and the rate of evolution of the reaction was studied in both cases as a function of antibody dose. Differences in the morphologic appearance of the reaction in these two cases were defined. It was demonstrated that there is no absolute requirement for a latent period in the production of PCA in guinea pig skin. Rather, latency in this system must be defined in terms of enhanced anaphylactic sensitization as a function of time between the administration of antibody and challenge with antigen. Quantitative comparisons between the amount of antibody needed to produce these two types of PCA reactions and the amount of antibody needed to produce anaphylaxis with soluble immune complexes suggest that the underlying mechanism of all three may be similar. Studies using carbamylated antibody with marked reduction in antigen-aggregating ability suggest that small immune complexes are involved in the production of PCA when antibody and antigen are injected separately. Therefore in analogy with the situation in which soluble complexes are used, where the active complex has been shown to contain two molecules of antibody, it is argued that PCA reactions involve the binding of antigen to at least two molecules of antibody present at adjacent sites on an effector cell. If this is so, then latency, as defined above, may be determined by a time-dependent reassociation of antibody and antigen on the cell surface until optimal configurations are attained.
Non-Complement-Fixing IgM Antibodies in the Guinea Pig
Authors: Linscott William D. , Hansen Stephen S.
Published: December 2022

The direct and indirect C1 fixation and transfer techniques were used to estimate the amounts of complement-fixing and non-complement-fixing IgM antibodies present in serial samples of serum taken from individual guinea pigs after immunization with rabbit erythrocytes, sheep erythrocytes, boiled sheep erythrocyte stroma, or bovine γ globulin. In most cases there was an increase in the relative proportion of non-complement-fixing IgM with time after immunization, and the ratio of non-complement-fixing antibody to complement-fixing antibody ranged from approximately 1 to greater than 10. Most antibody preparations contained several times as much non-complement-fixing IgM as complement-fixing IgM.
Prolonged Bursal Lymphocyte Depletion and Suppression of Antibody Formation Following Irradiation of the Bursa of Fabricius
Authors: Weber W. T. , Weidanz W. P.
Published: December 2022

Destruction of lymphoid follicles with lasting lymphocyte depletion was observed when the bursa of Fabricius of day-old chicks was exposed to 1000-r doses of x-radiation on the 1st and 7th day of post-embryonic life. Irradiated bursas did not reach more than 2% of the control bursa weights over a 90-day period of observation. The numbers of germinal follicles and plasma cells were decreased in the spleens and cecal tonsils of bursa-irradiated (BI) birds. In addition, BI birds demonstrated a diminished antibody-producing capability to selected antigens, and several of them showed a transient γG dysgammaglobulinemia. No histologic differences between the thymuses of bursa-irradiated and non-irradiated birds were detected.
Mechanism of Bacteriophage T4 Neutralization by Rabbit Immunoglobulin and Its Proteolytic Digestion Fragments
Authors: Stemke G. W.
Published: December 2022

Coliphage T4 is neutralized more efficiently by divalent than by univalent antibody fragments. The size of the univalent fragment is not of major significance. While first-order one-hit neutralization of T4 is obtained with divalent neutralizing agents, univalent agents neutralize normally with two-hit kinetics. However, a tail fiber amber mutant of T4, X4e, is neutralized with multitarget kinetics. A “decision” assay of the effect of univalent fragment on T4 likewise leads to one-hit kinetics. If dissociation of univalent fragment explains its lower activity as compared to divalent agents, it is not the kind of dissociation detectable by the “decision” assay.
Production of Anti-Hapten Antibodies by Several Classes of Lower Vertebrates
Authors: Leslie Gerrie A. , Clem L. William
Published: December 2022

Antibody against DNP was demonstrated in several classes of lower vertebrates immunized with DNP-protein conjugates. In many cases, the antibody could only be detected by the neutralization technique employing DNP-T4 phage; however, teleosts in general appeared capable of synthesizing rather high levels of antibody to DNP.
Determination of Antibody-Hapten Equilibrium Constants by an Ammonium Sulfate Precipitation Technique
Authors: Stupp Yehudit , Yoshida Takeshi , Paul William E.
Published: December 2022

The determination of the equilibrium constant of antibody-hapten interactions has led to important insights into the specificity and structure of antibodies (1) and into the nature and control of the immune response (2). The techniques presently used for measuring this constant all have certain disadvantages. Equilibrium dialysis (3), which must be regarded as the primary method, is relatively tedious to perform whereas other techniques, such as fluorescence quenching (4), require the use of purified antibody and of expensive equipment.

We now report the satisfactory determination of the average intrinsic association constant (K0) of hapten-antibody interaction on the basis of data obtained by precipitating antibody-hapten complexes with ammonium sulfate solutions. This represents an extension of the well-known Farr technique (5), which has previously been used to estimate the avidity of anti-bovine serum albumin antibody.
Model Systems for the Study of Drug Hypersensitivity
Authors: Hoffman Donald R. , Campbell Dan H.
Published: December 2022

Anti-aspiryl antibodies were produced in rabbits using three protein carriers. The method of inhibition of precipitation was used to determine the specificity of the antibody produced. ε-Aspiryl-aminocaproate was found to be the most effective inhibitor of those haptens tested, indicating that the combining hapten of the protein is ε-aspiryl-lysyl. Free aspirin and salicylates were poor inhibitors and did not combine with the antibody to a significant extent in equilibrium dialysis experiments. The ortho group in aspiryl and salicylyl amides was found to participate in the binding to antibody. The average binding constants of antibody globulins and specifically purified antibody were measured.
In Vitro Immune Responses of Rabbits with Persistent Adenovirus Type 5 Infection
Authors: Reddick Rhoda A. , Lefkowitz Stanley S.
Published: December 2022

Antibody responses of rabbits to adenovirus type 5 were studied. Serum antibodies were still present 12 months after the animals were given a single injection of infectious virus. Isolated lymphoid tissues from these animals were capable of synthesizing high antibody titers in vitro if exposed to viral antigen prior to cultivation. Antibody-producing cultures were maintained up to 120 days. Serum antibody titers of animals immunized with the soluble antigens declined rapidly. Synthesis of antibody could be inhibited by the addition of chloramphenicol and puromycin to the culture media. Phytohemagglutinin enhanced antibody synthesis of established antibody-producing cultures. The antibody species synthesized in vitro was primarily 7S although some 19S antibody was produced. Evidence for a persistent infection in the lymphoid tissues of rabbits given infectious virus was obtained by recovery of the virus from the tissues of the animals up to 12 months after the initial injection. The virus was also isolated from spleen and lymph node fragments maintained in vitro. This study indicates that persistent infection of the rabbit with adenovirus type 5 was responsible for the prolonged immune response.
Enhanced Humoral Immunity in Mice Infected with Attenuated Venezuelan Equine Encephalitis Virus
Authors: Howard R. J. , Craig C. P. , Trevino G. S. , Dougherty S. F. , Mergenhagen S. E.
Published: December 2022

Attenuated Venezuelan equine encephalitis virus (VEE) infection increased the antibody response to human γ globulin (HGG) in adult Balb/c mice. Mice infected 1 or 11 days before but not 29 days before immunization with 1 mg HGG showed increased rates of elimination of 131I-HGG and elevated hemagglutination titers to HGG. In addition, VEE infection prevented the induction of tolerance to HGG and converted a tolerance-inducing stimulus into an immunizing stimulus. However, VEE infection did not prolong the survival of skin allografts across weak or strong histocompatibility barriers.

Two to 6 days after infection, mice showed increased rates of clearance of colloidal carbon. This may have been related to the proliferation of reticuloendothelial cells, since VEE infection caused a proliferation of these cells in germfree mice. In addition, germfree mice infected with VEE had increased spleen weights, and germinal centers appeared in histologic sections of spleens. Increased γ globulin concentrations were seen in the sera of infected germfree mice, which could be attributed only partially to antivirus antibody. Thus, VEE behaves as an adjuvant for humoral antibody responses.
Spontaneous Thyroiditis in the Obese Strain of Chickens
Authors: Witebsky Ernest , Kite Joseph H. , Wick Georg , Cole Randall K.
Published: December 2022

Immunologic findings in the Obese Strain (OS) of chickens are described. This strain was obtained by selective breeding of individual OS chickens which comprised less than 1% of the original flock of the Cornell C-strain. At the present state of breeding about 80% to 90% of the individual chickens exhibit the OS characteristics. Tanned red blood cells from chicken, sheep and human coated with chicken thyroid extract are agglutinated by the majority of sera of OS chickens to various titers. Only chicken thyroid extract and no other chicken organ extract is capable of inhibiting the reaction. On the other hand, cells coated with human thyroid extract are essentially negative, showing the species-specificity of the chicken thyroglobulin antibodies. Precipitins can be easily demonstrated in the sera of OS chickens. By both methods (indirect hemagglutination and precipitation) the autoantibody nature of the thyroglobulin antibody can be demonstrated using thyroid material from the antibody-producing chicken itself. Upon sucrose gradient fractionation of OS chicken serum the highest hemagglutination titer appeared in the lighter antibody fraction and a lower one in the heavier antibody region.
Varying Specificity of Systemic Lupus Erythematosus Sera for Histone Fractions and a Periodatesensitive Antigen Associated with Histone
Authors: Stollar B. David
Published: December 2022

Three human systemic lupus erythematosus (SLE) sera reacted with free histones and showed varying selectivity for the histone fractions. Two sera were primarily reactive with F1 histone and one with F2a. Another serum appeared to react specifically with F2a2 histone, but the antigen which reacted with this serum was inactivated by periodate and was removed from the histone by electrophoresis. A fifth serum, which did not react with free DNA or free histone, reacted with histone-DNA complexes, but only when they were formed with the F2a2 histone; this component of histone antigen remained active after exposure to periodate and was recovered with homogeneous F2a2 protein on preparative electrophoresis. The sera also varied in their relative reactivities with free histone and nucleoprotein.
Myeloma Proteins with Anti-Hapten Antibody Activity: ε-2, 4-Dinitrophenyl Lysine Binding by the Protein Produced by Mouse Plasmacytoma MOPC-460
Authors: Jaffe Bernard M. , Eisen Herman N. , Simms Ernest S. , Potter Michael
Published: December 2022

Approximately 5% to 10% of mouse myeloma proteins react selectively with nitrophenyl ligands (1, 2), and nearly all of the reactive proteins are IgA immunoglobulins. Comparison of the binding properties and structural features of these proteins is therefore of some interest. The protein produced by mouse plasmacytoma MOPC-315 was previously shown to bind ε-DNP-l-lysine with an affinity (intrinsic equilibrium association constant) of 1 to 2 × 107 M-1 (1). In this preliminary study we show that another A-myeloma protein, produced by mouse plasmacytoma MOPC-460, has a markedly different affinity for the same ligand.

MOPC-460 was induced in an inbred, pedigreed Balb/c An N mouse by three intraperitoneal injections of mineral oil (Bayol F, Humble Oil and Refining Co., Bayonne, N. J.) (3). The tumor was maintained in serial transplant by subcutaneous passage every 4 weeks. Pooled sera, obtained by cardiac puncture of mice with large tumors, were stored at -20°C.
A Small Molecular Weight Permeability Factor in Guinea Pig Serum: Adsorption to Antigen-Antibody Aggregates
Authors: Movat Henry Z. , Treloar Mary P. , Takeuchi Yuko
Published: December 2022

The plasma kinin system is readily activated in vitro by glass and other surface active agents (1, 2). Davies and Lowe (3, 4) showed that in addition to anaphylatoxin a permeability factor, which is not suppressed by antihistamine, is activated when serum is incubated with immune precipitates. They demonstrated the possible role of complement in the generation of this factor (designated Pf/P), but failed to show its relationship to any known chemical mediator. We have presented evidence that guinea pig, rabbit and human serum can be activated by immune aggregates (5), and that such serum contains at least two distinct kinin-forming enzymes in addition to other permeability factors, presumably activators (6, 7). Activation of the kinin system by antigen-antibody complexes is of interest because it may have in vivo significance.

We have further investigated the properties of the vascular permeability enhancing material that becomes adsorbed on the immune aggregates.
Structural Localization of Isotypic Markers on Mouse Myeloma γG Globulins
Authors: Medgyesi G. A. , Gorini G. , Doria G.
Published: December 2022

Antibodies to the papain-sensitive antigenic determinants of mouse myeloma γG globulins were produced in guinea pigs. These antibodies could be used to identify isotypic markers on globulins of different subclasses. Immunochemical analyses of globulin fragments obtained by different proteolytic enzymes indicated that amino acid residues located both near to the site of papain cleavage and on the Fc participate in the expression of the papain-sensitive antigenic determinants.
Communications
Authors: Wang A. C. , Goodman J. W. , Fudenberg H. H.
Published: December 2022

The N-terminal residue on the heavy chains of five human IgA myeloma proteins was determined. Three of these heavy chains had PCA at their N-terminal end while the other two had flee N-terminal amino groups. The presence or absence of PCA at the N-terminal position of the a chain was not correlated with any of the following characteristics of the IgA molecules: a) antigenic subclass of the a chains; b) inherited anti-genic determinant of a chains, the so called Am(l) factor; and c) class of light chain associated with the a chain in the intact IgA molecule.
The Use of Egg Albumin Conjugates for the Purification of Antibody
Authors: Szafran Halina , Beiser Sam M. , Erlanger Bernard F.
Published: December 2022

A simple procedure is described for obtaining purified anti-hapten antibodies in good yield. The hapten was coupled to egg albumin in solution and the resulting heavily substituted conjugate was rendered insoluble by boiling in solution at pH 4.5. The insoluble product was used as an immunoabsorbent, the purified antibody being recovered by displacement with hapten.
An Active Interferon Inducer Obtained from Hemophilus Influenzae Type B
Authors: De Clercq Erik , Merigan Thomas C.
Published: December 2022

An extract was prepared from Hemophilus influenzae type b which induced significant interferon production both in vitro (human skin fibroblasts) and in vivo (mice). When compared to the endotoxin of Salmonella enteritidis the H. influenzae extract was 10 times more active in inducing interferon in human cells, 10,000 times more active in inducing cellular resistance to vesicular stomatitis virus in human cells and 300 to 1000 times less toxic in inducing lethality in mice and dermotoxicity in rabbits. The antiviral activity of the extract was not affected by ribonuclease, deoxyribonuclease or trypsin but was partially reduced by incubation at 25°C or 37°C. The chemical structure of the active substance of the H. influenzae extract has yet to be defined.
Studies on the Polylysine Immune Responder Gene. The Rank Order of Immunogenicity of Dinitrophenyl Conjugates of Basic Homopolyamino Acids in Guinea Pigs
Authors: Levine Bernard B.
Published: December 2022

The poly-l-lysine (PLL) gene appears to control an “antigen-processing step” permitting responder guinea pigs to make immune responses to hapten-PLL conjugates. This paper reports the finding of a rank order of immunogenicity of dinitrophenyl (DNP) conjugates of basic homopolyamino acids: poly-l-lysine τ; poly-l-arginine τ; poly-l-homarginine τ; poly-l-ornithine in both antibody formation and delayed hypersensitivity in responder guinea pigs. A similar rank order was observed for the hydrolysis of these conjugates by crystalline trypsin, except that polyarginine was hydrolyzed slightly faster than was polylysine. Complexing the ornithine and homarginine conjugates to bovine serum albumin considerably increased their immunogenicity. These data indicate that the same gene controls the immunogenicity of all four basic polyamino acids. The data suggest the hypothesis that the above rank order may reflect the specificity of interaction of these conjugates with the PLL gene-product.
Identification and Genetic Control of two Rabbit Immunoglobulin Allotypes at a Second Light Chain Locus, the c Locus
Authors: Gilman-Sachs Alice , Mage Rose G. , Young Glendowlyn O. , Alexander Cornelius , Dray Sheldon
Published: December 2022

Cross immunization of rabbits of different strains with IgG of varying electrophoretic mobility resulted in production of two precipitating allo-antibodies, which identified two IgG immunoglobulin light chain allotypes, c7 and c21. The c7 and c21 allotypes segregate as if controlled by allelic genes; however, progeny data of a few families suggest that the genes may instead be closely linked, i.e., pseudoalleles. The b and c light chain loci of immunoglobulins are not closely linked. The c locus is not linked to other immunoglobulin loci, i.e., the a heavy chain or f Ig A loci, nor to other serum protein loci, the Lpq low density lipoprotein and the Mtz α2 macroglobulin loci.
Lymphoid Precursors: Thymus Independent Antibody Production
Authors: Tyan Marvin L. , Herzenberg Leonard A. , Gibbs Priscilla R.
Published: December 2022

Thymectomized and nonoperated adult mice were lethally irradiated and given liver cells from embryos of various ages. The resultant chimeras were hyperimmunized with sheep erythrocytes, and cell suspensions from their spleens were used in a hemolytic plaque assay employing specific anti-allotype antisera as developing agents in order to distinguish between cells producing host and donor-type γG2a (7S) antibodies. It was found that lymphiod stem cells taken from embryos as early as the 10th day of gestation (i.e., before the appearance of the thymic rudiment) were able to produce γG2a anti-SRBC antibodies in thymectomized hosts. The number of donortype “developed” plaques found in the spleens of thymectomized hosts was approximately one-tenth that found in their nonoperated controls.

Sensitization of the hosts to SRBC before irradiation and the transfer of embryonic liver cells did not increase the number of donor-type PFC.
The Effects of Heterologous Antithymocyte Sera in Mice
Authors: Mandel Mark A. , DeCosse Jerome J.
Published: December 2022

The inoculation of Balb/c mice with 5 × 104 MPC 37 plasma cell tumor cells resulted in death of the animals in 8 to 9 weeks. Treatment of these mice with a single 0.5-ml dose of antithymocyte serum on the day of tumor inoculation resulted in an accelerated rate of tumor growth and death of all animals within 5 weeks. This rapid tumor growth effect was noted even when ATS was given 3 days prior to tumor inoculation. When mice given antithymocyte serum were injected intravenously 2 days later with 7.5 × 107 syngeneic adult spleen cells, a partial reversal of the rapid tumor growth was noted. Critical considerations in this reconstitution were the timing of the injection and number of cells administered. When tumor cells were incubated in ATS prior to their inoculation into mice, accelerated tumor growth did not result. These findings were corroborated by similar experiments carried out with a second tumor, MOPC 104E.

The relationship between tumor size and host serum γ2b immunoglobulin level was constant in mice that received tumor alone, tumor and ATS, and tumor, ATS, and then syngeneic spleen cells. Thus, treatment of the host did not seem to alter the functional capability of the tumor.

The accelerated tumor growth rate found in mice treated with ATS appears to be due to the immunosuppressive effects of this antiserum. A potential hazard of ATS is, therefore, that it lowers the ability of the host to cope with oncogenic challenges.
Enzyme Activation by Antibody
Authors: Suzuki T. , Pelichová H. , Cinader B.
Published: December 2022

Antisera to bovine pancreatic ribonuclease were fractionated on columns of DEAE Sephadex A-50 and the fractions were characterized by allotype content, electrophoretic mobility, the heterogeneity of heavy chains and the effect on enzyme activity.

Neutralizing antisera contained fractions which augmented ribonuclease activity in the presence of cyclic cytidylic or cyclic uridylic acid as substrate, but not in the presence of nucleic acid. Such antibodies were contained in a fraction P1 which could be separated as the first eluate from DEAE Sephadex A-50 and freed of small quantities of inhibiting antibody by rechromatography. The resulting fraction P1′ was less heterogeneous than other fractions and its heavy chains had unique mobilities and showed a minimal tendency to polymerize.

The properties of these activating antibodies differed from those previously obtained by immunization with polyalanine-polytyrosine ribonuclease. Some other fractions, all more heterogeneous than P1, also showed neutralization properties which differed from those found with unfractionated antisera. This was particularly marked with fractions P3–P6, which gave an initial decrease in enzyme activity as a function of increasing levels of antibody, and a secondary increase in enzyme activity with yet higher antibody concentrations.
Antiphagocytic Effects of Staphylococcal Protein A
Authors: Dossett John H. , Kronvall Göran , Williams Ralph C. , Quie Paul G.
Published: December 2022

Strains of Staphylococcus aureus possessing large amounts of protein A tended to resist phagocytosis more than strains that contain lesser amounts or none of this protein. Protein A extracted from S. aureus inhibited phagocytosis of both S. aureus and Escherichia coli in in vitro phagocytosis systems. The antiphagocytic effect was produced by nonspecific reaction of protein A with the Fc piece of γG globulin.
Effect of Disodium Cromoglycate on Certain Passive Cutaneous Anaphylactic Reactions
Authors: Lopez Manuel , Bloch Kurt J.
Published: December 2022

Disodium cromoglycate, in certain doses, completely inhibited the 48-hr PCA reaction in the rat prepared with homoeytotropic antibody and partially inhibited the 4-hr PCA reaction in rats prepared with IgGa antibodies. It is suggested that disodium cromoglyeate may inhibit the immunologic release of histamine and serotonin from rat mast cells regardless of the immunoglobulin class of the antibodies involved.
A Model for the Interaction of Cell Types in the Generation of Hemolytic Plaque-Forming Cells
Authors: Groves D. L. , Lever W. E. , Makinodan T.
Published: December 2022

The results of limiting-dilution and dose-response experiments involving the primary immune response of dispersed mouse spleen cells in Millipore diffusion chambers are presented. Sheep erythrocytes were employed as antigen and the immune response assessed by the enumeration of hemolytic plaque-forming cells using the classical Jerne technique. Limiting-dilution analysis demonstrated that the number of “response units” involved in initiating the immune response was a linear function of the spleen cell dose. The frequency of these response units in the antigen-stimulated spleen cell population was also estimated. Logarithmic dose-response curves dealing with the magnitude of the immune response as a function of spleen cell dose were found to be biphasic. At low doses of spleen cells the magnitude of the response was a nonlinear function of the cell dose, but this relationship abruptly became linear at higher spleen cell doses. A stochastic model for the initiation of the immune response based on the interaction of two cell types is proposed and this model is compatible with the findings of both limiting-dilution analysis and dose-response experiments. This model suggests that a response unit is formed by the multiplicative interaction of precursors of antibody-forming cells with antigen-reactive cells, the former cells becoming transformed into antibody-forming cells through such an interaction. That is, an individual antigen-reactive cell is able to interact with and influence the transformation of more than one precursor cell and the number of precursor cells transformed per antigen-reactive cell is a function of the spleen cell dose.
The Immunochemistry of Immune Tolerance
Authors: Baer Harold , Dawson Charles R. , Byck Joseph S. , Kurtz A. Peter
Published: December 2022

Immune tolerance was induced in guinea pigs to pentadecylcatechol and to cyclohexylmethylcatechol when oil solutions were injected subcutaneously. Structural features of these molecules associated with the induction and specificity of delayed contact sensitivity were operative in the induction and specificity of immune tolerance. These factors include the size and shape of the hydrocarbon side-chain which determine specificity, the presence of unsubstituted ring hydroxyl groups that permit quinone formation, and the availability of ring positions to permit coupling. Serum antibody could be found only occasionally either in sensitive or tolerant animals.
Chemical Modification of Human and Rabbit Antistaphylococcal γG Antibodies: Effect of Acetylation, Carbamylation and Amidination on Opsonic Activity
Authors: Messner R. P. , Parker C. W. , Williams R. C.
Published: December 2022

Opsonically active γG-globulins isolated from five patients with staphylococcal subacute bacterial endocarditis and two hyperimmunized rabbits were subjected to acetylation, carbamylation and amidination to determine if substitution of chemical groups in the γG molecule would result in loss of opsonic activity. The degree of chemical substitution was monitored by the ninhydrin reaction. In vitro opsonic capacity of all γG preparations tested decreased with progressive acetylation or carbamylation. No preparations were opsonically active after substitution of 50% of the free amino groups. The ability of the antibodies to react with staphylococcal antigens was not significantly diminished by these treatments. Both acetylation and carbamylation produced a marked increase in the net negative charge of γG globulin and significantly altered antigenic determinants of both light and heavy chains. Limited alteration of γG opsonins was obtained with ethylacetimidate which substitutes only at α and ε amino groups. Amidination of up to 87% of the free amino groups did not alter the charge of the γG preparations and resulted in only minor changes in antigenic determinants of the light and heavy chains. The ability of the antibody to combine with staphylococcal antigens was retained. The opsonic capacity of human γG was abolished by substitution of 65% of the free amino groups. A comparable degree of amidination of rabbit γG globulin had no significant effect on its opsonic activity.
Isolation of Molecules of Restricted Allotype from Antistreptococcal Polysaccharide Antibody
Authors: Rodkey L. Scott , Choi Tat K. , Nisonoff Alfred
Published: December 2022

Rabbits were immunized with pepsin-treated Streptococcus pyogenes, strain C-39. Two of thirty-five rabbits immunized responded with antibodies of limited electrophoretic heterogeneity, as indicated by zone electrophoresis and isoelectric focusing. Both of these rabbits were heterozygous, one at the a locus and the other at the b locus. Antibodies in the major peaks of the isoelectric focusing profile were refocused. The antibodies were then distributed in a very narrow range of isoelectric pH. In each case the protein in the peak had a distribution of allotypic specificities markedly different from that of the IgG of the same serum, with one specificity predominating. When light chains were isolated from the preparation heterozygous at the b locus, and subjected to isoelectric focusing, 99% of the chains in the major peak were of b5 specificity. In contrast, isoelectric focusing and refocusing of nonspecific IgG had no appreciable effect on the distribution of allotypic specificities.

The results provide support for the hypothesis that a substantial portion of certain antipolysaccharide antibody populations may be of monoclonal origin. The data also suggest that the isoelectric focusing technique may be applicable to the isolation of relatively homogeneous antibody populations on a preparative scale.
Macrophage Aggregation in Vitro: A Correlate of Delayed Hypersensitivity
Authors: Lolekha Somsak , Dray Sheldon , Gotoff Samuel P.
Published: December 2022

A macrophage aggregation test which uses peritoneal exudate cell suspensions (PEC) in tubes in the presence of antigen was found to correlate with cutaneous delayed hypersensitivity. Moreover, the supernatant fluids from lymphoid cell cultures from antigen sensitive animals caused aggregation of nonsensitive PEC in the presence of antigen. Thus, antigen leads to the release of a macrophage aggregation factor (MAF) from sensitive lymphoid cells in culture. By serial dilution, the MAF titers were determined for the supernatant fluids of various lymphoid cell cultures.
The Biologic Activities of Guinea Pig Antibodies
Authors: Sandberg Ann L. , Osler Abraham G. , Shin Hyun S. , Oliveira Benedito
Published: December 2022

Guinea pig γ1 immunoglobulins diminish the hemolytic activity in fresh serum when added as preformed immune precipitates. The loss in lytic potency is due primarily to interaction with the late C components beginning with C3. This fixation profile differs from that of the γ2 immunoglobulins which utilize all of the C components, whether they interact with antigen in the presence of C, or are incubated with C as preformed precipitates. Since the γ1 immunoglobulins do not cleave C3 in the presence of EDTA, it is suggested as one alternative, that a metal requiring enzyme other than the C4–C2 convertase may be involved in initiating C utilization by guinea pig γ1 immunoglobulins.
An In Vitro Measure of Cellular Immunity to Fibroma Virus
Authors: Tompkins W. A. F. , Adams Cathryn , Rawls W. E.
Published: December 2022

A modification of the macrophage migration inhibition test using infected monolayers as a source of antigen was applied to the in vitro study of cellular immunity to fibroma virus infection. The capacity of infected monolayers or infected cell homogenates to inhibit migration of peritoneal exudate cells from fibroma virus-infected rabbits paralleled the onset of a cutaneous delayed hypersensitivity response and resistance to re-infection in the intact animal. Studies using antigens prepared at different intervals after infection of cells with the virus suggest that the test is measuring specific antigen(s) associated with the cell surface and probably not a component of the intact infectious virion. Immune serum did not prevent the antigen from inhibiting migration of sensitized cells, but instead enhanced the inhibition. The data obtained demonstrate that the macrophage migration test has practical application in the study of cell-mediated immunity to animal virus infections.
Isoelectric Separation of Mouse Immunoglobulins
Authors: Murgita Robert A. , Vas Stephen I.
Published: December 2022

New classes of immunoglobulins have recently been characterized, and major classes divided into subclasses. Proteins of the known classes are structurally distinct and some differences in biologic properties are known, but the general significance of these immunoglobulins is as yet not clear. Prior to a study of the biologic functions of the individual classes of immunoglobulins, techniques must be available which will consistently yield relatively large amounts of pure immunoglobulin. Several methods for purifying γ globulin have been described (1–3), but many of them have limitations that make them unsatisfactory for obtaining a consistently pure product from sizeable quantities of serum (4, 5). It is unlikely that a one-step procedure can efficiently purify large amounts of protein and therefore it is necessary to select a combination of techniques that will work best to accomplish purification. The majority of methods for purification use separation on DEAE-cellulose as a major step in the procedure (6); others use zone electrophoresis in conjunction with ion exchange chromatography in a two-step procedure (7).
Correlation of Charge Properties of Human Reaginic Antibodies with Charge of the Corresponding Allergens
Authors: Underdown Brian J. , Goodfriend Lawrence
Published: December 2022

Reaginic (skin-sensitizing) antibodies in the sera of ragweed allergic individuals show considerable heterogeneity in elution properties when chromatographed on the anion exchanger, DEAE-Sephadex (1, 2). Evidence was obtained (3) that the chromatographic heterogeneity was primarily due to charge and not to size differences between the reaginic antibody molecules. A basis for charge heterogeneity of antibody molecules in general was suggested by the recent finding of Sela and Mozes (4) that specific rabbit antibodies vary in their overall charge depending (inversely) on the net charge of the corresponding immunogen. The availability of two purified ragweed pollen allergens, antigen Ra.3 (5) and antigen E (6), which differ in net charge as well as other physicochemical and immunologic properties, made it possible to investigate whether a similar relationship might obtain for human reagins formed to the allergenic constituents of ragweed pollen.
A Sensitive Neutralization Test for the Adeno-Associated Satellite Viruses
Authors: Boucher D. Wark , Parks Wade P. , Melnick Joseph L.
Published: December 2022

A highly sensitive neutralization test for the four satellite serotypes was developed by selecting particularly effective combinations of adenovirus helper and cell system. Neutralizing antibody titers were measured by inhibition of satellite replication as assayed in terms of complement-fixation-producing units (CFPU) of virus. Neutralizing antibody measured by this method was more type-specific, and the antibody titers were much higher than those obtained by the direct complement-fixing antibody test. Homotypic neutralizing antibody titers of 103 were common, and in some instances titers as high as 106 were observed. The neutralizing activity against satellite virus was found to reside in the λ globulin fraction of post-infection human and post-immunization rabbit sera. The frequency of individuals with neutralizing antibodies but without CF antibodies suggests that the CF antibody titer falls off more rapidly than the neutralizing titer. Both neutralization and CF tests revealed significant intertypic crossing, especially between types 2 and 3.
Polyclonal 13S Human γG Immunoglobulin
Authors: Heimer Ralph , Martinez Jose , Abruzzo John
Published: December 2022

Polyclonal 13S γG immunoglobulin, isolated from the serum of a patient with dermatomyositis, dissociated to 6.5S material possessing anti-γG activity. The Fc but not the Fab fragment, prepared from the 13.3S material, formed soluble complexes with autologous and isologous γG immunoglobulin. Aggregation rather than an antigen-antibody reaction appeared to be involved in the formation of the γG complex.
Immunologic Properties of Glucose Oxidase
Authors: Green Ira , Inman John K.
Published: December 2022

Glucose oxidase, an anzyme obtained from Aspergillus niger, is a potent antigen. Guinea pigs and rabbits immunized with this antigen in complete Freund's adjuvant produced high serum concentrations of precipitating antibody. In addition, heat-inactivated enzyme and apoenzyme can be used to detect cellular immunity in the guinea pigs.

Glucose oxidase retains its enzymatic activity when bound to antibody, thus allowing direct visualization of the latter upon incubation with a suitable mixture of substrate and chromogenic reagent. Plasma cells producing antibody to the enzyme were visualized in smears and in tissue sections by sequentially reacting fixed preparations with glucose oxidase and chromogenic reagent. Anti-DNP antibody-producing cells elicited by immunization of guinea pigs with dinitrophenylated-ovalbumin were also visualized in a similar manner by employing dinitrophenylated glucose oxidase as a test antigen.
Effect of Antigenic Stimulation on the Specificity of Antibody Produced by Rabbits Immunized with Bacteriophage MS-2
Authors: Krueger R. G.
Published: December 2022

Repeated immunization of rabbits with the RNA coliphage MS-2 resulted in the appearance of an increasing amount of neutralizing antibodies that showed an increased rate of association (i.e., affinity) with the homologous phage antigen. There was a progressive narrowing of the specificity of the antibodies produced after each antigenic stimulation as evidenced by a decrease in the extent of cross-reactions observed with related heterologous RNA phages. These results are discussed in relation to the nature of the antibody response and the antigenic structure of phage MS-2.
A Rapid, Quantitative Fluorescence Assay for Cell Damage by Cytotoxic Antibodies
Authors: Edidin Michael
Published: December 2022

Our laboratory recently (1) described a fluorochromatic assay for cell damage by cytotoxic alloantibody, which was based on retention of fluorescein by intact cells after its hydrolysis from fluorescein diacetate esters (FDA) (2). Although the method gave reproducible results in our hands, it was slow and always subject to error, since traces of complement used in the reaction were capable of hydrolyzing FDA, thus yielding erroneously high values of cell viability. This was not satisfactory and accordingly we sought another approach to differential fluorescent staining of cells, which would still allow machine-reading of the assays. Rather than looking for other compounds that were enzymatically changed from non-fluorescent to fluorescent, we chose to screen a number of fluorescent probes (3), compounds whose fluorescence is enhanced when bound to protein or nucleic acids. Such compounds were initially screened by observing fresh and heat-damaged lymphocytes, bathed in solutions of various probes, with the fluorescence microscope.