bluuebunny/crossref_metadata_2025 huggingface dataset:
Univalence of Half-Molecules of Rabbit Antibody
Authors: Hong Richard , Palmer Jerry L. , Nisonoff Alfred
Published: January 2023

Dissociation of specifically purified rabbit antiovalbumin into half-molecules, followed by recombination at neutral pH, resulted in a 6 S product, 81% of which was precipitable by antigen. The sedimentation coefficient, precipitability, and shape of the precipitin curve indicated that most of the molecules in the product were bivalent.

When the recombination was carried out in the presence of a large excess of dissociated normal γ-globulin, the product did not form specific precipitates with antigen, but specifically inhibited the reaction of ovalbumin with untreated anti-ovalbumin. The results are consistent with univalence of half-molecules of antibody and the previously demonstrated capacity of half-molecules of anti-ovalbumin to combine with halfmolecules of normal γ-globulin, while retaining the integrity of antibody combining sites.

A procedure is described for specific purification of rabbit anti-ovalbumin and anti-bovine γ-globulin.
A Study of the Antigenic Relationship between Human Measles Virus and Canine Distemper Virus
Authors: Roberts J. A.
Published: January 2023

The antigenic relationship between human measles virus and canine distemper virus has been analysed.

Neutralization titrations of antisera produced by dogs in response to homologous and heterologous inoculation producedures indicated that the capsids of the viruses have similar antigens, but that there is no common identical antigen.

Adsorption of antibody onto, and elution from, preparations of purified virus indicated that at least part of the antibody produced during the heterologous secondary response comprises antibody molecules that can react with both viruses.
Immune Degradation in Passively Sensitized Mice
Authors: Terres Geronimo , Sorem Gail L.
Published: January 2023

Mice were injected with a constant amount of radioactive iodine (I*)-rabbit anti-bovine serum albumin (BSA) and concomitantly with different amounts of BSA such that the zones of antigen excess, equivalence and antibody excess were included. It was shown that both the amount and the rate of immune degradation of I*-rabbit anti-BSA were dependent on the amount of antigen injected. At equivalence the onset of immune degradation was rapid whereas in the zone of antigen excess the onset was delayed. In antibody excess the rate of degradation only slightly exceeded that of the controls. The degradation of I125-labeled antibody and of I131-labeled antigen were followed simultaneously. A constant amount of I125-RγG (rabbit γ-globulin) was injected with different amounts of I131-BSA representing combinations in antigen excess, equivalence and antibody excess. In each case there was an apparent delay in the onset of immune degradation of the antibody, whereas the immune degradation of the antigen began immediately except in antibody excess. Possible explanations for these results were discussed. The data support the idea of constant rearrangement of complexes in vivo such that the instantaneous rate of degradation is a function of the actual ratio of antigen-to-antibody remaining. The data also suggest that more than one type of antibody is involved in accelerated degradation and retention of labeled antigen.
Common Antigenic Determinants of Penicillin G, Cephalothin and 6-Aminopenicillanic Acid in Rabbits
Authors: Brandriss Michael W. , Smith James W. , Steinman Harry G.
Published: January 2023

The antigenic properties of penicillin G, cephalothin and 6-aminopenicillanic acid were studied in rabbits. The benzylpenicilloyl group appeared to be the major determinant of the penicillin G antigen, in confirmation of earlier reports, but we could not confirm that it was the only determinant involved. Cephalothin and 6-aminopenicillanic acid formed cross-reacting antigens with penicillin G and the cross-reaction was inhibited by benzylpenicilloyl hapten. This suggests the possibility that cephalothin and 6-aminopenicillanic acid become antigenic in vivo through direct conjugation of the β-lactam carbon to native protein and that the penicillins might follow the same pathway.
Induction of Immunologic Unresponsiveness in Chickens with Avian Serum Proteins
Authors: Tempelis C. H.
Published: January 2023

Intraperitoneal injections of 1.0 or more mg of turkey γ-globulin (TGG) into “wet” chicks resulted in partial or complete inhibition of precipitin production at 6 weeks of age. Persistence of complete tolerance was evident up to 12 weeks in the group receiving 50 mg TGG. Partial tolerance was present at 22 weeks in those receiving 100 mg TGG. Greater amounts of goose γ-globulin (GGG) were necessary to produce even a partial state of tolerance. This correlates with the phylogenetic relationship of the two proteins to the chicken. These experiments confirmed that the duration of immunologic tolerance was related to the dose of the antigen administered at birth and that chickens are made tolerant more easily to avian than mammalian serum proteins. Chickens produce good quantities of precipitating antibodies to GGG but not to TGG.
Antigenic Properties of Human Mucous Membranes
Authors: Bonstein Herbert S. , Mazow Jack B. , Arbesman Carl E. , Rose Noel R.
Published: January 2023

Human respiratory mucosa crude extract was fractionated by ammonium sulfate precipitation, starch block electrophoresis and agar gel electrophoresis. The serologic reactions of these fractions were studied by means of immunoelectrophoresis, double diffusion in agar and tanned cell hemagglutination. The respiratory mucosa fractions precipitated by 0 to 1.34 M ammonium sulfate (Fraction I), 1.34 to 2.00 M ammonium sulfate (Fraction II), and 2.00 to 3.00 M ammonium sulfate (Fraction III) all produced precipitation with specific antiserum to bronchial mucosa. Lung was similarly fractionated and showed visible precipitation lines in Fraction II and Fraction III using the same bronchial mucosa antiserum. By means of double diffusion in agar all the lines formed by the various fractions of respiratory mucosa showed reactions of identity. The faint lines formed by fractions of lung were partially identical to those formed by fractions of respiratory mucosa. Tanned cell hemagglutination tests were positive in highest titers with Fractions I and II of respiratory mucosa, Fraction II giving a lower titer. The remaining fractions did not react in hemagglutination tests. Inhibition of hemagglutination of Fraction I-coated cells and Fraction II-coated cells was shown by Fractions I, II and III of respiratory mucosa. Fraction III inhibited agglutination of Fraction II-coated cells in greater dilution than did Fraction II itself. No significant inhibition was demonstrated by extracts of lung, esophagus, or kidney. Different levels of the respiratory tract differ quantitatively in their content of the antigens represented in Fraction II.
Use of Polistes Venom in Petrolatum: Arlacel Repositories to Immunize against Yellow Jacket Wasp-Sting Allergy
Authors: Loveless Mary Hewitt , Nall Thomas M.
Published: January 2023

Summary and Conclusions

Twenty-four persons with extreme allergy toward yellow jacket wasp stings were immunized safely with single, yearly, subcutaneous repositories of emulsified heterologous venom from the polistes wasp, there being no significant adverse developments on treatment day and no longterm sequelae such as abscess, cyst or neoplasm in the next 3 to 4 years. By contrast, control groups with identical allergy exhibited two adverse responses among 111 single-visit courses of fluid venom prepared from the homologous wasp or from polistes wasps. Effectiveness of therapy was judged by the clinical response to later stingings, of which 60 took place by plan in the hospital and four in the field by accident. Aside from trivial focal developments in 12 instances, only two significant adverse episodes were engendered, one in a repository-treated man and one after an immunization with homologous fluid venom. Subsequent repetition of the challenge found both men fully tolerant, one having taken a booster of fluid polistes venom in the meantime while the second merely rested. It was deduced that factors other than specificity differences in the two immunizing venoms probably had been responsible for the clinical failure after repository. The 64 challenges involved one to four insects and post-treatment periods that ranged from 9 days to 21 weeks for courses of fluid venom, and from 15 to 27 weeks for depot therapy. All but a few of the sting tested subjects have been transferred to intermittent sting-therapy to maintain their demonstrated immunity and are now receiving 2-min stingings from 3 to 4 wasps—polistes in cold months, yellow jacket wasps in summer—at intervals approaching a year. Threshold tests of skin and eye may afford a clue, before immunization, to the patient's tolerance for injected venom. After therapy, there is a small but statistically significant elevation in requirement, due to induced blocking antibody. As to the indication for specific prophylaxis in sting allergy, probably only those victims whose next encounter threatens disaster merit immunization, in view of the implied long-term risk of using foreign proteins parenterally. Obviously, only the simplest preparations and the smallest effective amounts are justified in any case, viz., venom as opposed to whole body extracts, and a single annual treatment as opposed to weekly and monthly injections over protracted periods.
Differences in the Electrophoretic Mobility of Antibody from Inbred Strains of Mice
Authors: Fahey John L. , Barth Werner , Ovary Zoltan
Published: January 2023

The electrophoretic mobilities of precipitating and skin-sensitizing antibodies were investigated in eight strains of mice. The precipitating antibodies produced by C57BL/6JN mice were slow migrating, while BALC/c and other strains of mice produced relatively rapidly migrating antibodies. Skin-sensitizing antibodies differed in electrophoretic mobility in the same manner as precipitating antibodies.

The mobility difference between mouse strains was confirmed for antibodies against three different antigens. Genetic factors evidently influence the electrophoretic mobility of antibody in mice.

These observations also show that some of the factors determining differences in electrophoretic mobility of immunoglobulin molecules are independent of antigen-binding (antibody) sites and the skin-sensitizing site.
Antigen Excess in Guinea Pig Anaphylaxis
Authors: Frick O. L. , Liacopoulos P. , Raffel Sidney
Published: January 2023

Systemic anaphylaxis in passively sensitized guinea pigs failed to occur when the animals were challenged with massive doses (0.5 to 1.0 g) of protein antigen intracardially. In the isolated sensitized guinea pig lung, bronchospasm was diminished or absent when large amounts of antigen were used for perfusion. PCA in guinea pig skin passively sensitized with rabbit anti-DNP sera was markedly reduced when large concentrations of divalent DNP haptens (bis-DNP lysine and bis-DNP ornithine) were employed.

These observations suggest that the inhibition of anaphylaxis by large concentrations of antigen may depend upon the formation in vivo of “nontoxic” complexes of composition Ag2Ab.
Interference with Establishment of Immunologic Tolerance to Shigella Antigens in Neonatal Mice by Specific Antiserum
Authors: Friedman Herman
Published: January 2023

Administration of hyperimmune mouse anti-Shigella serum to newborn mice receiving a tolerance-inducing inoculum of Shigella soluble antigen (SSA) interferes with establishment of tolerance. An interval of 1 or 2 days between injection of antigen at birth and administration of antiserum results in only partial interference with tolerance, while an interval of several days or more results in no demonstrable effect. γ-Globulin containing fractions prepared from hyperimmune mouse serum is also capable of preventing induction of tolerance when administered at birth or shortly thereafter. Normal mouse serum, heterologous antiserum and hyperimmune anti-Shigella serum absorbed with Shigella organisms are ineffective. Administration of specific antiserum several weeks following induction of tolerance does not permanently “break” tolerance to Shigella. It is postulated that specific antibody to the tolerance-inducing Shigella antigen may combine with and/or neutralize the antigen in vivo in the newborn mouse so that no antigen remains free to induce and maintain tolerance.
An Analysis of Certain Factors Related to the Quantitation of Immunoelectrophoresis
Authors: Russell Warren J.
Published: January 2023

Factors involved in determining antigen concentration by relating immunoelectrophoretic precipitin arc position to the origins of antigen and antibody have been analyzed using refined immunologic systems. The data have been statistically analyzed, results evaluated and recommendations set forth.

A linear relationship is found to exist for both albumin and γG-globulin when the p value (diffusivity ratio) is plotted against the log antigen concentration. A departure from linearity was not detected by the technique employed, even though the diffusion coefficients of the reactants were different. It was found that the p value difference must exceed 0.075 for albumin-antialbumin and 0.059 for γG-globulin-anti-γG-globulin before any significant differences can be declared at the 0.05 level.

A comparison between experimentally determined equivalence and theoretical equivalence p values has been made and results indicate that the observed differences are most likely due to a decrease in peak antigen concentration when the antigen is moved in the electrical field.

On the basis of the data in this report, an experimental design is presented for determining unknown antigen concentrations by immunoelectrophoresis. This protocol specifies the number of samples needed to establish significant differences. Confidence intervals for various p values are given.
Reactions of Localizing Anti-Kidney Antibodies with Tissues of Various Species
Authors: Gery Igal , Yagi Yasuo , Pressman David
Published: January 2023

Reactions of localizing anti-kidney antibodies with kidney (and liver) of homologous and heterologous species were studied by the “paired label technique,” using mixtures of I125-labeled antibody and I131-labeled normal globulin.

The kidney localizing activity of rabbit antirat kidney antibodies is largely removed by absorption with rat kidney sediment and is removed to various lesser extents by prior absorption with kidney sediments of other species.

Anti-kidney antibodies formed in rabbits against sheep kidney and antibodies formed in goats and guinea pigs against rabbit kidney localize in vivo to a small extent in kidneys of rats.

Cross-localization reactions become much more apparent by the use of antibodies purified by absorption on and elution from kidney and liver sediments. Purified antibody preparations react differently with various tissue antigens, indicating that the purification procedure selects certain populations of anti-kidney antibodies out of the much more heterogeneous original mixture. Each population cross-reacts preferentially with the tissue antigens of the species used for purification.

A small degree of cross-reaction was observed even with kidney of the species used for antibody production. This was shown only after purification on this tissue sediment.

The cross-reaction of anti-rat kidney antibody with kidneys of heterologous species may be useful in the characterization of the antigens responsible for the specific antibody localization in the rat kidney.
Circulatory Collapse in Anaphylaxis and Histamine Toxicity in Mice
Authors: Bergman R. K. , Munoz J.
Published: January 2023

Active anaphylaxis, passive anaphylaxis and histamine were all found to cause a highly significant rise in hematocrit values over prechallenge levels in Swiss-Webster strain (CFW) mice. The rise in hematocrit values indicated a loss of intravascular fluid sufficient to lower the average blood volumes of these groups 23 to 38% within 5 min after challenge. Injecting plasma extenders, i.e., physiologic saline or 6% dextranphysiologic saline, into mice during the first hour after challenge protected them from death, except for normal mice challenged with 24.0 mg of histamine. Injection of plasma extenders into mice receiving this massive histamine challenge gave no beneficial effects whatsoever and even seemed to be deleterious.

This study strongly indicates that anaphylactic and histamine death in hypersensitive mice is due to circulatory collapse which is brought about by loss of blood volume into the extravascular space and for which the animal is unable to compensate. The histamine-sensitizing factor (HSF) from B. pertussis appears either to augment the blood volume-depleting effects of anaphylactic and histamine shock or to interfere with the vascular compensatory mechanism. The increase of histamine sensitivity produced by dichloroisoproterenol (DCI) seems to have similar mechanisms.
Enumeration of Antibody-Forming Cells in the Peripheral Blood of Immunized Rabbits
Authors: Kearney R. , Halliday W. J.
Published: January 2023

Rabbits were injected with sheep erythrocytes, and their blood leukocytes were separated at intervals and examined by the agar plaque technique for cells producing specific antibody.

In a typical experiment, no active cells appeared after the first injection, but a second stimulus consistently produced antibody-forming cells in considerable numbers (up to several hundred/ml of blood). These cells first appeared on the 2nd day after the second antigen injection, reached a maximum on the 3rd day and rapidly declined. Subsequent injections gave rise to smaller responses.

An attempt was made to estimate the efficiency of the separation procedure.

Splenectomy of the rabbit had no significant effect on the occurrence of antibody-forming cells in the blood.
Cross-Reactions among γ-Globulins of Various Species in Indirect Immunofluorescence
Authors: Henle Gertrude , Henle Werner
Published: January 2023

Several preparations of fluorescein isothiocyanate-conjugated rabbit antibodies to guinea pig or human γ-globulin fractions were found to be capable of staining the characteristic mumps virus inclusions in infected HeLa cells previously sensitized with virus-specific antibodies derived from heterologous animal species. Brilliant immunofluorescence was elicited by labeled antibodies to heterologous γ-globulins only when the antiviral sera used for the primary stage of the reaction were used undiluted, or at the most in 10-fold dilution. The rabbit anti-guinea pig γ-globulin sera from which effective fluorescein conjugates were prepared yielded from one to three lines of precipitation in Ouchterlony tests with sera from several heterologous species with the exception of rabbits, whereas those sera from which strictly species-specific conjugates were obtained produced no precipitates with heterologous sera. Cross-reacting antisera to guinea pig γ-globulin also reduced the viral neutralizing capacity of human poliomyelitis convalescent serum. No attempts were made to identify the cross-reacting serum component(s). Such broadly reactive anti-γ-globulins might be useful for the indirect technique of immunofluorescence in that they overcome the usually observed species limitation of this method.
Effect of Bacillus Calmette-Guérin on the Levels of Acid Phosphatase, Lysozyme and Cathepsin in Rabbit Alveolar Macrophages
Authors: Heise E. R. , Myrvik Q. N. , Leake E. S.
Published: January 2023

A study has been made of the time course and dose-response curves of lysozyme, acid phosphatase and cathepsin in rabbit alveolar macrophages following the injection of bacillus Calmette-Guérin (BCG) by the intratracheal route. Significant elevation in the levels of these acid hydrolases was observed within 3 to 7 days following the injection of BCG. The increase in enzyme levels was observed to be a dose-related response.
An Agar Interaction in Immunodiffusion
Authors: Hokama Y. , Coleman Monroe K. , Riley Richard F.
Published: January 2023

The diffusion constant of C-reactive protein determined by the method of Allison and Humphrey is not the same for all samples of pathologic serum, indicating that C-reactive protein may exist in different sera in different states of aggregation. It is unlikely that it is strongly associated in native serum with β-lipoprotein.

Results of manipulating the ionic milieu and the nature of the supporting medium in which diffusion was carried out, indicated that C-reactive protein (CRP) shows a strong interaction with the charged groups of agar. The anomalous behavior of CRP on immunoelectrophoresis may result in part from this interaction.

The relevance of these observations to the quantitative determination of C-reactive protein (and probably some other antigens) by immunodiffusion techniques in agar were pointed out.
Studies on Mouse Antibodies
Authors: Adler Frank L.
Published: January 2023

Summary and Conclusions

The immune response of mice to one or more intraperitoneal injections of sheep red cells has been studied. A 25- to 100-fold variation in the dose of antigen that was injected into normal mice failed to have any significant effect on the formation of agglutinins and lysins. The kinetics of hemolysin and agglutinin formation differed, primarily because of the superior agglutinating activity of 7 S antibody, which appeared 24 hr after 19 S antibody and continued to increase as the titer of the latter dropped. While early lysins and agglutinins were ME-sensitive β-globulins, those appearing later were 7 S γ-globulins, among which the lysins appeared to possess greater electrophoretic mobility. The production of 19 S lysin in the primary response appeared to occur in two phases of which the early one was characterized by rapid rates of rise and decline, while the second phase was one of prolonged production at a slowly decreasing rate. Reinjection of antigen caused a rise of 19 S antibody in the continued presence of 7 S antibody which was similar to that following the first injection.

A class of 7 S lysins and agglutinins, of γ1 and γ2 mobilities, was found to be sensitive to ME. Such antibodies were encountered during the early phases of the 7 S antibody formation in primary and secondary responses. In contrast to 19 S antibodies, which lost all serologic activity after ME treatment, these sensitive 7 S antibodies fully maintained their combining ability.
Estimation of the Homogeneity of Purified Ragweed Antigens
Authors: Pruzansky Jacob J. , Patterson Roy
Published: January 2023

The antigenic homogeneity of three purified ragweed antigens has been studied by an antiglobulin coprecipitation technique using human antiserum. They were approximately 84, 67 and 64% homogeneous as determined by this procedure. Smaller amounts of other antigens were also present. Only one major precipitin band was found, and it was common to all three preparations. The utility and the limitations of the coprecipitation procedure were considered. The occurrence and significance of multiphasic binding curves in heterogeneous systems were discussed with the aid of a model consisting of a mixture of purified antigens and homologous antibodies.
Antibody Responses of Rhesus (Macaca Mulatta) Monkeys Experimentally Infected with Coxsackieviruses of Group B and Group A, Type 9
Authors: Schmidt Nathalie J. , Dennis Juanita , Lennette Edwin H. , Ho Helen H. , Shinomoto Tak T.
Published: January 2023

Neutralizing, complement-fixing (CF), precipitating and hemagglutination-inhibiting (HI) antibody responses were studied in rhesus monkeys experimentally infected by oral administration of coxsackievirus types B1, B3, B4, B5, B6 and A9. Antibody responses were studied both in initial infections and in successive infections with viral heterotypes. Neutralizing antibody responses were seen to be rapid and type specific; in successive infections with viral heterotypes antibody elicited by previous infections was often recalled, but antibody was not produced to viral types other than those with which the animals had experienced an infection. Complement-fixing antibody responses were type specific in initial infections, but subsequent infections served to broaden the antibody spectrum; not only was antibody recalled to viral types with which the animals had previously been infected, but CF antibody was produced to viral types with which the animals had ostensibly had no previous experience. CF antibody responses were also accelerated in successive infections. The monkeys produced both specific and group-reactive precipitating antibodies, the latter to viral types with which the animals had not been infected. As in the case of CF antibodies, the spectrum of group precipitating antibodies was broadened in successive infections. Specific precipitating antibody was produced only to the infecting viral type and was of short duration. HI antibody produced in response to infections with coxsackievirus types B1, B3 and B5 was type specific and tended to parallel neutralizing antibody in temporal appearance and persistence.
Studies with Rhesus Monkey Antisera to Human γM-Globulins (γM)
Authors: Mackenzie M. R. , Deutsch H. F.
Published: January 2023

Serologic differences between γM-globulin antibodies such as Rh saline agglutinins, the α- and β-isohemogglutinins and cold and heterophile agglutinins have been demonstrated by the use of rhesus monkey antisera to individual Rh saline agglutinins and to Waldenström γM-globulins. Differences in the γM-globulin antibodies within an individual human sera also exist.

Rhesus monkey antisera to γM-globulins give multiple precipitin bands with normal human serum or with isolated γM-globulins when the reactions are carried out in agar gel. This further indicates the antigenic complexity of these proteins.

Immunoelectrophoretic studies in agar indicate that the primary factors in human serum responsible for the precipitin reactions with the monkey anti-Rh saline agglutinins are γM-globulins. Starch gel immunoelectrophoretic experiments confirm this and further indicate that other proteins are present in Type O sera that react with monkey antibody to γM-globulins.
Frequency Distribution of α, γ, κ and λ Polypeptide Chains in Human Lymphoid Tissues
Authors: Bernier George M. , Cebra John J.
Published: January 2023

Fluorescent antibody preparations of different colors and specificities have been used to determine the qualitative and quantitative distribution of four kinds of immunoglobulin polypeptide chains in human splenic tissue. Single cells were found to contain one kind of heavy chain and one kind of light chain, but rarely, if at all, did two kinds of light chain (κ and λ) coexist in a single cell. Similarly, the heavy chains, α and γ, appeared to be mutually exclusive. The relative frequency of the two kinds of light chains paralleled the relative serum concentrations of these moieties. Cells containing γ chains were slightly more frequent than cells containing α chains, but not to the extent anticipated by the relative serum concentrations of these two immunoglobulins.
An Immunogenetic Study of the Rabbit Agglutinator Factor (Anti-Antibody)
Authors: Cohen Carl , Tissot Robert G.
Published: January 2023

Serum factors, called agglutinators, have been described which are capable of causing the agglutination of rabbit erythrocytes specifically sensitized with certain incomplete rabbit isoantibody. Of 13 different sensitized cell systems, 5 were capable of detecting the agglutinator factor. One of these systems detected an agglutinator factor which appears to be present as the result of the action of the recessive genes ag/ag.
Association of Immunologic Determination with the Lymphocyte Fraction of the Peritoneal Fluid Cell Population
Authors: Weiler Ivan Jeanne , Weiler Eberhardt
Published: January 2023

Two immune peritoneal fluid cell populations, determined with respect to different antibody specificities (bacteriophages T6 and T7), were fractionated in parallel. The lymphocyte fraction of specificity T6 (LyT6) was then combined with the macrophage-neutrophil fraction of the other specificity (M-NT7), and vice versa. The two heterospecifically recombined cell populations were each injected into a group of irradiated mice. The antibody titers of specificities T6 and T7 in each group were then related to the proportions of each of the respective cell types injected into the host animals.

Mice receiving the combination LyT6/M-NT7 produced more antibody of specificity T6 than of T7; mice receiving recombined LyT7/M-NT6 produced more T7 antibody. It is concluded that immunologic determination is associated with the lymphocyte fraction of the peritoneal fluid cell population.
Conversion of Nonimmune Spleen Cells to Antibody-Forming Cells by Rna: Strain Specificity of the Response
Authors: Cohen Edward P. , Newcomb Richard W. , Crosby Louise K.
Published: January 2023

Spleen cells from nonimmunized mice were converted to antibody-forming cells by an RNA extract from the spleens of immunized mice. The conversion took place in the presence of trypsin, pronase, both α and β-amylase and DNase, but was blocked by chloramphenicol and ribonuclease. RNA extracted from the spleens of mice immunized with an unrelated antigen was ineffectual, and the addition of RNA from nonimmunized mice to the active preparation reduced the number of antibody-forming cells. The conversion was found to be partially strain-specific. Separation of total cellular RNA revealed most of the active material to be in the 8 to 12 S fraction.
Studies on Immunity in Experimental Leptospirosis: the Immunogenicity of Leptospira Icterohemorrhagiae Attenuated by γ-Irradiation
Authors: Hubbert William T. , Miller James N.
Published: January 2023

Leptospira icterohemorrhagiae, γ-irradiated with 70,000 r or greater, failed to multiply in vitro, were rendered noninfectious for guinea pigs, yet maintained their motility in Stuart's medium for 2 to 10 days, the time varying inversely with the irradiation dosage. L. icterohemorrhagiae, γ-irradiated with 100,000 r or 1,000,000 r, retained their ability to induce agglutination-lysis antibody in both hyperimmunized rabbits and guinea pigs. Of 12 guinea pigs immunized with L icterohemorrhagiae, γ-irradiated with 100,000 r, 11 were protected against both symptomatic and latent renal infection as measured by the absence of positive blood and kidney cultures following challenge. This was in contrast to the formalinkilled, sonically disrupted and water-soluble vaccines which failed to provide adequate clinical protection. Little or no agglutinin response resulted following immunization with the four vaccines.
Immunochemical Properties of Human γA Isoagglutinin
Authors: Ishizaka Kimishige , Ishizaka Teruko , Lee Evelyn H.
Published: January 2023

The three immunoglobulins, i.e., γG, γA and γM, were isolated from the sera of type O humans, who were immunized with A substance. γA-globulin was fractionated into polymer and monomer types. The effect of reduction with mercaptoethanol and alkylation by iodoacetate on antigen-binding activity of anti-A antibodies were studied. The results indicated that the stability of the activity to the treatment was different depending on the immunoglobulins with which the anti-A antibody was associated; the susceptibility of both polymer and monomer types γA-antibodies was intermediate between that of γG- and γM-antibodies. Upon reduction and alkylation, the polymer type γA-antibody was dissociated to subunits containing antigen-combining site(s). It was also found that a part of the antigen-combining activity remained after reduction and alkylation of γM-antibody. Evidence was presented that the activity was associated with subunits of the antibody. Reduction of γM-antibody followed by dialysis against saline resulted in reassociation of subunits to form fast-sedimenting molecules having agglutinating activity.
Immunochemical and Genetic Studies of Two Distinct γG-Immunoglobulins in BALB/c Mice
Authors: Dray Sheldon , Potter Michael , Lieberman Rose
Published: January 2023

Twenty-seven different immunoglobulins prepared from sera and urine of BALB/c mice with plasma cell tumors were tested with a variety of mouse allotype antisera reacting with immunoglobulins of normal BALB/c mice. None of the antisera obtained from 15 different donor-recipient combinations reacted in Ouchterlony precipitin tests with the one γM- and three γA-immunoglobulins nor with the ten λ and two κ light polypeptide chains from mice with plasma cell tumors. Several of the antisera gave weak precipitin reactions near the antigen well with some of the five immunoglobulins designated γF; these reactions were interpreted as the probable result of contamination with small amounts of immunoglobulins from normal plasma cells.

The antisera distinguished between two classes of γG-immunoglobulins designated γG-Be1 and γG-Be2. Some antisera gave dense precipitin bands with the three tumor-derived γG-Be2 immunoglobulins as well as the corresponding γG-Be2 immunoglobulin in normal serum but not with the three tumor-derived γG-Be1 immunoglobulins. Similarly, some antisera gave dense precipitin bands with the three tumor-derived γG-Be1 immunoglobulins as well as the corresponding γG-Be1 immunoglobulin in normal serum but not with the three tumor-derived γG-Be2 immunoglobulins. The allotypic specificities which distinguish between γG-Be1 and γG-Be2 (designated a1′ and a1") were found to be associated with the papain Fc-fragment, a part of the heavy polypeptide chain.

Antisera specific for γG-Be1 and γG-Be2 (anti-a1′ and anti-a1″) were used to test F2 progeny for independent assortment or linkage. The a1′ and a1″ allotypic specificities were either both present or both absent in the same F2 progeny indicating close linkage in their genetic control.
Studies in Immunization
Authors: Salvaggio John E. , Flax Martin H. , Leskowitz Sidney
Published: January 2023

Beryllium, a known granuloma-producing agent in man, has been substituted for mycobacteria in Freund's adjuvant and its effects on antibody production and delayed hypersensitivity to bovine serum albumin have been observed in rabbits and guinea pigs.

Animals inoculated with beryllium adjuvant emulsions exhibited only a moderate increase in antibody production to the antigen when compared with a control group given bovine serum albumin (BSA) in incomplete Freund's adjuvant. However, they developed virtually no delayed hypersensitivity to BSA when compared to a third group given BSA in complete Freund's adjuvant (with mycobacteria).

Histologic study showed marked similarities in the cellular response of the local injection sites of beryllium and mycobacterial adjuvants. However, the granulomata associated with beryllium were not as extensively disseminated or as rich in epithelioid cells as those associated with mycobacterial adjuvants.
A Study of Phase Variation in Coxiella Burneti Employing Hemagglutination Tests
Authors: Bakemeier Richard F.
Published: January 2023

Hemagglutination tests with normal and tannic acid-treated RBC exposed to ultrasonicates of Coxiella burneti have been employed in a study of the phase variation of these organisms. The results are consistent with the hypothesis, initially derived from complement fixation studies, that Phase I C. burneti contain a component(s), with the RBC-adsorption characteristics of polysaccharides, which is markedly deficient, although probably present, in purified suspensions of Phase II organisms. Rickettsiae of both phases appear to contain protein antigens reacting with anti-Phase II sera.
Preparation of Antiserum to Enteroviruses in Large Animals
Authors: Hampil Bettylee , Melnick Joseph L. , Wallis Craig , Brown Russell W. , Braye Edward T. , Adams Raymond R.
Published: January 2023

The usefulness of five species of large domestic animals for the preparation of enterovirus immune serum was investigated systematically, using horses, cattle, sheep, goats and swine and five representative enterovirus strains, namely, poliovirus 1, coxsackievirus A9, coxsackievirus B3, and echoviruses 4 and 11.

Monkey kidney tissue culture fluids of the prototype strains were treated with molar MgCl2 to preserve maximal viral infectivity and antigenicity, and concentrated tenfold by the Mg-Al carbonate method, for use as immunizing antigens. The results of serum neutralization tests showed that horses, cattle, sheep and goats produce serum of useful titer against all five viruses, and large volumes of antisera were collected. Swine responded to poliovirus 1 but not to the other four viruses. The horse appears to be the animal of choice in terms of the height of the antibody response and the closely similar titers among individual animals of each group. Booster doses, administered 21 to 28 weeks after completion of primary immunization with four doses of antigen, were effective in elevating the serum antibody titers to levels equal to or above the highest titers previously observed. Recommendations are proposed for a practical scheme to yield antiserum of high titer (10,000 range) in volumes as large as 25 L, from as few as two animals.

A problem of fundamental significance was emphasized by tests performed on the echovirus 4 antisera. These sera, though high in neutralizing antibody titer against the DuToit strain of echovirus 4, were essentially negative when tested against the prototype virus (Pesascek) which had been used as the immunizing antigen. This paradoxical situation has been studied and is discussed further in a separate publication (19).
The Specificity and Interaction with Poliovirus of an Inhibitory Bovine Serum
Authors: Pagano Joseph S. , Gilden Raymond V. , Sedwick W. David
Published: January 2023

Bovine sera capable of inhibiting plaque formation of poliovirus may be highly selective in their action. A bovine serum that strongly inhibited plaque formation of the CHAT type 1 attenuated poliovirus had little effect on the 24 other type 1 strains tested, whether attenuated or virulent. An inhibitory factor, located in the 19 S serum globulin fraction by gel filtration with Sephadex G-200, did not affect replication of the sensitive virus, but it did initially neutralize virus in a cell-free system according to first order kinetics. The neutralized virus was reactivated at pH's below 5.0.

In correlating the inhibition in plaque reduction tests with the effects of cell-free neutralization, three results were observed: a) virus sensitive in both systems, b) virus nonsensitive in plaque reduction tests but sensitive to direct incubation with inhibitory serum and c) virus relatively nonsensitive in both systems.

In experiments with sequential neutralization, the addition of fresh bovine serum to mixtures containing virus previously exposed to bovine serum did not cause neutralization of the remaining infectious virus. However, type 1 antiserum did neutralize additional virus, although the amount neutralized and the rate of the second reaction were both somewhat diminished. Conversely, bovine serum added to complexes of virus and type 1 antiserum did not neutralize the residual virus.
Partial Separation of Skin-Sensitizing Antibody from Antibodies Which Bind Antigens in vitro
Authors: Kopp William L. , Bilbo Richard E. , Reed Charles E.
Published: January 2023

Gel filtration and density-gradient sedimentation partially separated the immunoglobulins in sera of atopic persons so that the distribution of antibodies to ragweed and house dust antigens could be studied. Binding of radioiodine-labeled antigens was assayed by the Farr technique and skin-sensitizing antibody by passive transfer. Binding of labeled house dust antigen correlated with the distributions of both IgM and IgG. Binding of ragweed varied. In some sera maximal binding correlated with IgM, in others with IgG, and in still others with both immunoglobulins. In three sera which incidentally contained antibody to BSA, binding appeared to be associated only with IgG. The distribution of binding of ragweed and house dust correlated poorly with the distribution of skin-sensitizing antibody. While a large amount of binding of these antigens by IgG might obscure a small amount of binding by IgA, most of the antibodies which bind ragweed and house dust antigens in vitro are IgM and IgG.
Characterization of the Antibody for Vibrio Fetus Endotoxin in Sera of Normal and V. Fetus Infected Cattle
Authors: Winter A. J.
Published: January 2023

Antibody for Vibrio fetus endotoxin has been demonstrated in sera from normal cattle of different ages. The V. fetus antibody resembles the “natural antibodies” described for other endotoxins in its specificity and in the uniformity of its distribution and concentration in serum. Antibody active in a passive hemagglutination reaction with lipopolysaccharide sensitized cells was associated almost exclusively with the γ1-macroglobulin component. Precipitating activity was localized mainly in the lower molecular weight globulins. Certain characteristics of the precipitins suggest that they may be γ1A-globulins. The presence of the venereal variety of V. fetus in bulls brought about no detectable change in quality or quantity of antibody. Venereal vibriosis in heifers led to the formation of serum precipitins more heat-stable than those in normal sera. In one animal which had aborted from vibriosis in the 6th month of gestation, precipitating and hemagglutinating activities were divided between the γ2- and γ1-macroglobulins.
The Adjuvant Action of Latex Particulate Carriers
Authors: Litwin Stephen D. , Singer Jacques M.
Published: January 2023

Polystyrene latex particles, with human γ-globulin adsorbed to their surface, acted as an adjuvant to antibody production. Several features of this adjuvancy were studied. It was found that the enhanced antibody response was present during the primary immune response but could not be demonstrated on secondary stimulation. No adjuvant effect occurred unless the FII and latex particle were in direct contact with one another. The antibody production during the immune response was dose-dependent on the amount of protein antigen used and was unaffected by very wide variations in the number of particulate carriers utilized, or the diameter of the particulate carrier (range from 500 Å to 13,000 Å). Antigenicity was not increased when fewer protein molecules were adsorbed per latex particle. During this adjuvant-induced primary antibody response the antibody population was initially composed of 19 S and 7 S molecules, followed later by 7 S antibody.

The data obtained suggested that the adjuvant effect of the human FII latex particulate carrier system was due to alteration of the configuration of the protein rather than being a result of changes in phagocytic uptake and handling.
Chemical and Physical Variables Affecting the Properties of Fluorescein Isothiocyanate and Its Protein Conjugates
Authors: Klugerman Maxwell R.
Published: January 2023

The effect of pH, time, temperature, and buffer systems on solutions of fluorescein isothiocyanate and its protein conjugates was examined fluorometrically. The fluorescence of the isothiocyanate and of the protein conjugate was maximal at about pH 8.7 and pH 10.7, respectively. The stability of their fluorescence above pH 7 was affected adversely by increases in pH or temperature. The protein conjugate, however, showed maximum stability at pH 10.5 and above. The type of buffer (carbonate, phosphate, borate, Tris and barbiturate) did not affect the fluorescence of the free dye significantly. On the other hand, increasing the molarity of the buffer caused a decrease in stability of fluorescence of the free dye but did not seriously affect the fluorescence of the conjugate.

The absorbance of fluorescein isothiocyanate and its conjugates increased with increasing pH. In 1 N NaOH, the effect of pH variation on absorbance was minimized.

By increasing the pH or temperature of the reaction mixture during conjugation, fluorescein isothiocyanate reacted more readily with the protein. Conditions may be selected to obtain the desired degree of label with short conjugation periods. Conjugation of a bovine anti-Brucella abortus globulin sample for 30 min at pH 9.45 and room temperature was as effective as conjugating at pH 8.75 for 18 hr at 5°C. No apparent loss of biologic activity as the result of conjugation was observed.

The binding of fluorescein isothiocyanate to protein resulted in a loss of fluorescence efficiency, the extent of loss being dependent upon the pH of measurement.

Existing procedures in fluorescent antibody studies should be reevaluated on the basis of the variables discussed.
Structural Characteristics of Red Cell Autoantibodies
Authors: Eyster M. Elaine , Nachman Ralph L. , Christenson William N. , Engle Ralph L.
Published: January 2023

The direct Coombs' reaction has been studied in 18 cases of autoimmune hemolytic anemia not associated with elevated cold agglutinin titers. The erythrocyte coating substance was γG-globulin alone in five patients, γG-globulin in combination with complement in nine, and complement alone in four. It has been demonstrated that the heavy chain and both types of light chains are involved in the γG-antiglobulin reaction. Variation in the strength of direct antiglobulin reactions with specific antisera to Bence Jones proteins of type K or type L indicated that the autoantibodies in nine of these patients were a heterogeneous group of γG-globulin molecules. Possible sites of complement fixation to the red cell membrane and to the γ-globulin molecule are discussed in relation to these findings.
Arthus Type Inflammation with Rabbit γM Antibody
Authors: Tada Tomio , Ishizaka Kimishige
Published: January 2023

Rabbit γM antibody against human type A substance gave precipitin reactions in vitro and induced reversed Arthus type of inflammation in guinea pigs which had received A substance. Preformed complexes composed of A substance and γM antibody also induced the allergic inflammation in normal guinea pig skin. On a weight basis, the minimum dose of the γM antibody for the induction of hemorrhage in reversed Arthus reactions was two to four times as much as the dose of the γG antibody. In the reactions by the preformed complexes, a similar dose of γM and γG antibody in the form of complexes with A substance induced comparable skin reactions.
Role of Interferon during Viremia
Authors: Baron Samuel , Buckler Charles E. , McCloskey Richard V. , Kirschstein Ruth L.
Published: January 2023

Mice, chickens, hamsters, rats and monkeys were studied for induction of circulating interferon following viremias resulting from infection or intravenous injection of diverse viruses. Most combinations of animal species and virus types resulted in early production of serum interferon, although not all such combinations were successful. These results indicate that circulating interferon is a rather general response by animals during the viremic phase of virus infection. Interferon was observed to leave the serum in vivo at a rate of about thirtyfold/hr. Using this rate it can be conservatively estimated that a total of 100,000 units of circulating interferon are produced during many of the viremias of mice.
Adjuvant Properties of Bacterial Cell Wall Mucopeptides
Authors: Holton Juanita Bird , Schwab John H.
Published: January 2023

Antibody production in rabbits against BSA, as measured by passive hemagglutination, gel diffusion and skin reaction, is enhanced when injected with saline suspensions of mucopeptide derived from the cell walls of β-hemolytic streptococci. The mucopeptide is more effective than comparable doses of cell walls or soluble cell extract. The best antibody response is obtained when the interval between mucopeptide and antigen injections is less than 24 hr. The adjuvant effect is local rather than systemic, since the antigen must be injected into the same skin site as mucopeptide. The mucopeptide has no effect on the immunoglobulin category in which antibody is found.