bluuebunny/crossref_metadata_2025 huggingface dataset:
The Incompetence of the Hamster Cheek Pouch Membrane as a Mechanical Barrier to the Movement of Tritiated E. Coli B3 and Tritiated T4 Bacteriophage
Authors: Shepro David , Cohen Paul S. , Kula Nora
Published: January 2023

Tritium labeled Escherichia coli B3 and tritium labeled T4 phage were injected into the hamster cheek pouch or hamster ear, the latter site serving as a control. Eighty per cent of the B3 and 98% of the T4 drained from the pouch within 72 hr. Comparable amounts of B3 and T4 escaped from the ear within the same period of time. These data indicate that the tissues of the cheek pouch do not act as a barrier to antigens the size of E. coli and T4 phage. The observation that the hamster produced phage specific antibodies following injections into the cheek pouch lends additional support to this view. The degree of antibody production was much lower in the cheek pouch-injected animals than in the controls. Based upon the above evidence, a theoretical explanation of the cheek pouch as a privileged site for homografts is here presented.
Purification and Characterization of the Hemagglutinin Present in Potatoes
Authors: Marinkovich Vincent A.
Published: January 2023

A procedure is described for the isolation of the hemagglutinin found in potatoes. Evidence is presented that this procedure results in a highly purified preparation.

The lectin is a nonspecific agglutinin with optimal activity at 4°C, and is reversibly bound to the red cells during agglutination. It is a basic glycoprotein with a sedimentation constant of 2.1 S, containing 12.5% nitrogen and 5.2% carbohydrate. A 0.1% solution of the lectin has a specific absorbance (K2780 Å) of 0.962 per mg-cm, and an agglutination dilution titer of 1:104. Thus agglutination is detectable at a lectin concentration as low as 0.1 γ/ml.

The lectin is shown to contain only one type of sugar, tentatively identified as arabinose. Amino acid analysis shows a preponderance of acidic amino over basic amino acids even though the lectin behaves as a basic molecule. This discrepancy is not easily explained by hydrolytic loss alone. The alternative possibility is presented that the sugar molecule is attached to the protein moiety by ester linkage to the free carboxyl groups of aspartyl and glutamyl residues, thereby mitigating their contribution to the net charge of the molecule.

The mechanism involved in the agglutinating activity of the potato lectin is briefly discussed.
Characterization of Early 19 S and Late 7 S Immunoglobulins in the Mouse
Authors: Kim Yoon Berm , Bradley S. Gaylen , Watson Dennis W.
Published: January 2023

Biphasic responses and differing rates of neutralizing activity of early and late sera from BALB/Sy mice given single and multiple injections of actinophage indicated two types of antibodies in phage neutralization.

By the use of physical and immunochemical techniques the neutralizing activity of early serum was found to be associated with a γ1-immunoglobulin of the 19 S class. The late serum contained mostly a γ2-immunoglobulin of the 7 S class.

The early 19 S antibody was inactivated by 2-mercaptoethanol in contrast to the stability of the late 7 S γ2 antibodies.

Immunoelectrophoretic patterns of pooled mouse sera were typical of rabbit and human patterns. In the mouse, however, mercaptoethanol dissociates the so-called β2M into a new component with the mobility of, but antigenically distinct from, the γ2-immunoglobulin.

In the mouse, the 19 S immunoglobulin is not related to the classical β2M line but is associated with the fast γ2-globulin and appears antigenically identical.

The implication of the apparent close relationship of these two classes of immunoglobulin is discussed in relation to the mechanism of their production.
The Effect of X-Ray Irradiation upon the Sequence of Immune Globulins Following Initial Immunization in the Rabbit
Authors: Robbins John , Smith Richard T.
Published: January 2023

The effect of x-ray irradiation upon the sequential formation of immune globulins was studied. With increasing doses of total body irradiation, a delay in the appearance of γ2-globulin antibody was observed, whereas only a slight delay in the timing of the initial detection and minimal diminution in the over-all titer of γ1M-macroglobulin antibody was noted. After lethal irradiation during which various portions of the animal's lymphoid tissue was shielded, a prolonged period of exclusive γ1M-macroglobulin synthesis was demonstrated. The restoration of immunopoietic tissue during the shielding procedures could be accomplished with the cells of the spleen, appendix, hind limb and peripheral blood. Simultaneous immunization of shielded and normal tissue after lethal irradiation demonstrated a normal sequential formation of antibodies to that antigen used to inject the shielded tissue but specific inhibition of γ2-globulin antibody to that antigen used to inject irradiated tissue. Various hypotheses which attempt to explain the relative resistance of the γ1M-macroglobulin phase of the immune response as compared to the γ2-globulin phase to total body irradiation were discussed.
Globulins on NZB Mouse Erythrocytes
Authors: Norins Leslie C. , Holmes Margaret C.
Published: January 2023

Using four rabbit antisera specially absorbed and then tested for hemagglutinating activity against various mouse erythrocytes and by immunoelectrophoresis against mouse serum, the major coating of direct Coombs positive NZB mouse erythrocytes has been found to be a classical γ-globulin. It is considered that this confirms the resemblance of the autoimmune hemolytic anemia of NZB mice to the corresponding human disease.
The Fate of Radioactive Bovine γ-Globulin during the Primary Antibody Response in the Mouse
Authors: Sweet Lawrence C. , Abrams Gerald D. , Johnson Arthur G.
Published: January 2023

The blood clearance rate and tissue distribution of I125 bovine γ-globulin (BGG) in the mouse following a single intraperitoneal injection with and without the stimulus of endotoxin have been studied. The circulatory fate of BGG was found to be similar to that previously described in rabbits. Whole organ counts after initiation of antibody synthesis revealed the greatest levels of radioactivity in the kidneys followed by liver, spleen and lungs. There was a redistribution of residual tissue radioactivity with time such that the relative amount in the kidneys tended to increase and that in the lungs to decrease while the relative amount in the liver and spleen remained essentially constant. Tissue autoradiograms revealed a selective localization of radioactive material in the reactive follicular centers of the spleen and mesenteric lymph nodes as well as the glomeruli of the kidneys. Antibody titers in mice receiving I125 BGG alone were higher than anticipated and were equal to those in mice stimulated also with endotoxin. No difference in antigen distribution was seen under these conditions in endotoxin-stimulated mice.
Biologic Properties of Polynucleotides
Authors: Yachnin Stanley , Ruthenberg Janet M.
Published: January 2023

Of a variety of naturally occurring and synthetic polynucleotides, only polyinosinic acid (poly I) is capable of evoking vascular permeability in guinea pig skin. This property of poly I is abolished by hydrogen-bonded helix formation with polyadenylic or polycytidylic acids, a maneuver which also destroys the capacity of poly I to interact with the complement system. The capacity of poly I to evoke vascular permeability is not affected by antihistamine. This biologic property of poly I supports previous evidence indicating that poly I can function as an activator of the first component of complement.
The Nature of the Antigen-Antibody Complexes which Initiate Anaphylactic Reactions
Authors: Levine Bernard B.
Published: January 2023

A divalent benzylpenicilloyl (BPO) hapten and several homologous multivalent BPO haptens of widely differing molecular sizes were compared quantitatively with regard to their abilities to evoke PCA reactions in guinea pigs passively sensitized with threshold, optimal and excess amounts of rabbit anti-BPO antibodies.

These haptens were compared also with regard to their abilities to specifically precipitate rabbit anti-BPO sera, and to evoke passive Arthus reactions.

It was found that equimolar concentrations (and not equal weight concentrations) of the divalent and of the multivalent haptens were equally effective in evoking PCA reactions throughout the dose-response curve. In contrast, equal weight concentrations of the multivalent haptens (and not equimolar concentrations) were found to be equally effective in precipitating anti-BPO sera, and in evoking passive direct Arthus reactions in guinea pigs sensitized with rabbit anti-BPO sera. The divalent hapten was virtually ineffective in precipitating anti-BPO antibodies present in these sera, and failed to evoke passive direct Arthus reactions.

These results are interpreted as demonstrating, at least in this system, that, whereas the precipitin reaction and the passive direct Arthus reaction depend on the formation of large cross-linked complexes, the PCA reaction in guinea pigs is initiated by the simple bridging of two (or a small integer (n)) firmly fixed anaphylactic antibody molecules by multivalent haptens to form comparatively simple immune complexes of the composition H1Abn.
Evaluation of Electronic Measurements of Hemagglutination for Quantitative Studies
Authors: Gibbs Mary B. , Dreyfus Joseph C. , Aguilu Luis A.
Published: January 2023

A method has been developed whereby the model B Coulter counter can be employed for counting free cells present in agglutinated samples. The method is valid over the entire range of agglutination and a wide range of serum concentrations. Instrument log probit assay curves are identical in slope and position to those obtained by hemocytometry. The method is reliable with anti-A and anti-B agglutination but has not been applied to measurements of agglutination in other blood group systems.
Antimycobacterial Activity of Tuberculostatic Factor on Intracellular Bacilli
Authors: Kochan Ivan , Smith Linda
Published: January 2023

This study demonstrated that the tuberculostatic factor of human serum cannot exert its effect on intracellular bacilli in macrophages collected from normal, tuberculin-negative guinea pigs. It was found, however, that infected, immune macrophages in the presence of tuberculostatic human serum showed significantly inhibited bacillary multiplication. Since infected, immune cells maintained in a bacillary growth-supporting medium did not inhibit the multiplication, it was concluded that the intracellular inhibition in the presence of human serum was caused by tuberculostatic factor. Once tuberculostatic factor was introduced into phagocytic cells with sensitized bacilli, inhibition took place in the absence of the antimycobacterial factor in tissue culture medium.

The results suggest that the permeability of “immune” macrophages is different from that of normal cells. This permeability of cells collected from vaccinated animals might be attributed to their state of hypersensitivity, which is triggered by the intracellular bacilli or their products.
Experimental Sensitization to Vitamin E (Tocopherol) in Guinea Pigs
Authors: Lipton Murray M.
Published: January 2023

The delayed or tuberculin type of sensitization has been produced in guinea pigs (as shown by skin test) using the natural compound vitamin E (d, α-tocopherol) incorporated into complete Freund's adjuvant.

D, α-tocopherol or dl, α-tocopherol have been shown to be sensitizing when injected into the skin in quantities of 0.01 to 0.02 ml (approximately 10 to 20 mg). The skin site acts both as a sensitizing site and a skin test site on primary inoculation.

A vitamin E-like compound has been demonstrated immunologically in tubercle bacilli. This is indirectly observed by skin testing with pure α-tocopherol animals which have been injected with Freund's complete adjuvent less the tocopherol.

Although the guinea pig is constantly coming in contact by the alimentary route with tocopherol in its food, this does not desensitize the animal.
Serologic Differentiation of Trachoma Strains and Other Agents of the Psittacosis - Lymphogranuloma Venereum-Trachoma Group with the Aid of the Direct Fluorescent Antibody Method
Authors: Katzenelson E. , Bernkopf H.
Published: January 2023

The serologic relationship of various strains of the psittacosis-lymphogranuloma venereum-trachoma group of agents was examined with the aid of rabbit immune sera and antigens partially purified with fluorocarbon or diethylaminoethyl cellulose. The direct fluorescent antibody technique was employed. Major sharing of antigens between trachoma and inclusion conjuctivitis (TRIC) agents and two strains of lymphogranuloma venereum was demonstrated. No cross-reactions were observed between these strains and a strain of psittacosis. By the use of cross-absorbed immune sera, a subdivision of TRIC agents was obtained according to which strains Dari and Camal, isolated in Israel, and strains Bour and T'ang fell in Group I and strains Asgh, SA-1 and G-17 in Group II. The strain of inclusion conjunctivitis IC Cal-3 showed cross-reactions with both groups of trachoma and, in addition, some specificity of its own. With the aid of cross-absorbed rabbit immune sera it was possible to group trachoma agents directly from infected yolk sac smears.
Estimation of Molecular Size of Complement Components by Sephadex Chromatography
Authors: Borsos Tibor , Rapp Herbert J.
Published: January 2023

The molecular weights of C′1, C′2 and C′4 (the first, second and fourth components of complement) in a number of species were estimated by studying their behavior in gel filtration. The molecular weight found for C′4 was about 180,000 and for C′2 about 150,000. C′1 is a considerably larger molecule than C′4 or C′2, for it coincided with the high molecular weight fraction obtained from serum by gel filtration. Using intermediate products and purified components of guinea pig C′, we have demonstrated the presence of C′1, C′2 and C′4 in the sera of a wide variety of mammals.
Gm Genes and γG-Globulin Synthesis in the Human Fetus
Authors: Mårtensson L. , Fudenberg H. H.
Published: January 2023

By means of highly specific typing systems for genetic γ-globulin factors, evidence has been obtained that the human fetus may synthesize small amounts of γG-globulin. A comparison of the Gm(a) and Gm(b) titers of cord sera with the titers of sera from adults suggests that the two genes, Gma and Gmb, do not synthesize the same relative amounts of γ-globulin at different stages of ontogenetic development. Some, but not all, of the maternal anti-Gm's that sediment as 7 S γ-globulin in density gradient experiments are also detectable in the fetal circulation.
Identification of the Species of Origin of Cells by Mixed Hemadsorption: A Mixed Antiglobulin Reaction Applied to Monolayer Cell Cultures
Authors: Espmark J. åke , Fagraeus Astrid
Published: January 2023

The use of mixed hemadsorption for testing species identity is outlined. The test, which is essentially a mixed anti-globulin reaction, is performed directly on cell sheets in monolayer cultures. The sensitivity is high if it is assumed that the foreign cells appear in the form of colonies. The combination of the method with a disc test in pure cell cultures permits a satisfactory quantitation of titers of anti-cellular sera. Some advantages of the method are discussed.
Inhibition of the Development of Delayed Hypersensitivity by Treatment with Antiserum
Authors: Crowle Alfred J. , Hu C. C.
Published: January 2023

Daily i.p. injections in mice of homologous or heterologous antiserum to chicken ovalbumin performed during the period of initial sensitization specifically suppressed development and hastened waning of immediate and delayed hypersensitivities to this antigen. This suppressive effect was considerably more effective against delayed hypersensitivity than against immediate. Intensive antiserum treatments usually were employed to elicit this type of suppression, but it could be induced with as little as 1.2 µl of antiserum given in 12 daily equal aliquots, or by a single injection of nonisogenic immune lymphoid cells.

Immunologic suppression by antiserum treatments affects production of antibodies but does not interfere with the initial events of antigen recognition necessary for development of anamnestic responsiveness.

These findings indicate that although immediate and delayed hypersensitivities may be different mechanistically they cannot be considered independent of each other, for the presence or absence of one can influence development of the other. The implications of this conclusion for interpreting such general phenomena as antiseruminduced resistance to autoimmune diseases, immunologic enhancement, the pathogenesis of certain infectious diseases, tolerance and mechanisms of adjuvant activity are discussed.
Autoinhibition of Rheumatoid Factor Reactivity Demonstrated by a Block Hemagglutination Procedure
Authors: Oreskes Irwin , Plotz Charles M.
Published: January 2023

Various rheumatoid factor (RF) sera were titrated with human γ-globulin (HGG) coated tanned sheep cells at six cell concentrations ranging from 0.125% to 2.0%. It was found that in 36 of 63 sera tested there was an abnormally large decrease in titer with increasing cell concentration. Double logarithmic plots of titers vs. cell concentrations were linear. While linearity with a slope of -1.0 was to be expected, it was found that these 36 sera tested yielded slopes greater than -1.0.

Addition of aggregated HGG to a -1.0 slope serum caused a rise in slope. Removal of autologous 7 S HGG, by diethylaminoethyl (DEAE) column chromatography, from a high slope serum lowered the slope. Addition of 7 S HGG, prepared either from a high slope serum or from certain normal sera, to a -1.0 slope serum served to raise the slope. Partial removal of RF by adsorption of a low slope serum also caused an increased slope. Heating at 56°C for 30 min did not affect slope values.

These results suggest that the high slope effect is primarily due to autoinhibitory heat stable components present in the 7 S HGG fraction of most rheumatoid factor sera.
The Complement Fixation Reaction for Measurement of Immunogenic Antigens in Formaldehyde-Inactivated Poliomyelitis Vaccine
Authors: Macmorine H. G. , Wardlaw A. C. , Weber J. C. W.
Published: January 2023

A method for measuring complement fixation (CF) antigens in formaldehyde-inactivated poliomyelitis vaccine has been described in detail. The antisera used were absorbed with heated polioviruses to make the test specific for the types I, II and III D-antigens. A high degree of type specificity has been demonstrated and statistical analyses have indicated that replicate assays are reproducible within narrow limits.

The ability of trace amounts of anticomplementary substances to produce significant shifts in the end point of the titration has been clearly demonstrated. The levels of these substances can be reduced by dialysis against Carbowax and by ultracentrifugation procedures which also effectively concentrate the D-antigen.

A variety of experiments were carried out, to investigate correlation between CF measurements of the D-antigen and antigenic response in chicks. These included the yield of antigen following concentration and purification of vaccines, loss of antigen during formaldehydeinactivation and loss on long term storage at refrigerator temperature. In the latter there was evidence that the loss of potency was somewhat greater when measured by chick tests than by CF but the two methods showed an encouraging degree of correlation. It is suggested that CF testing with anti-D sera can provide useful data for accurate standardization of poliomyelitis vaccines.
Studies on the Interrelationship of Cold Environment, Immunity and Resistance to Infection
Authors: Northey William T.
Published: January 2023

Immunochemical studies were conducted on sera from rabbits to investigate the qualitative and quantitative aspects of the immune response under conditions of “hypothermia.” Rabbits have been subjected to lowered environmental temperature (4°C), shaved of their pelage and immunized with various protein antigens. Sera representing different antigens and conditions of cold exposure which were obtained from varying periods during each of the studies have been qualitatively assayed for antibody response by Ouchterlony gel diffusion, immunoelectrophoresis, starch gel electrophoresis and paper electrophoresis. Quantitative analyses included “ring” precipitin titrations, quantitative gel diffusion analysis and microquantitative precipitin analysis. In all of the studies the responses of the “cold exposed” animals have been compared with those of non-cold exposed animals treated in an identical manner, except for maintenance of the lower environmental temperature.

An evaluation of the data concerning the effect of “cold exposure” on the antibody response indicates that any “differences” attributable to “cold” per se from both the qualitative and quantitative aspects are of insufficient magnitude to indicate that cold exposure appreciably affects the antibody response of rabbits.
Agglutination of Latex Particles without Antigen Adsorption
Authors: Donaldson Paul , Pennington Karen
Published: January 2023

Latex particles were agglutinated during combination of antiserum with antigen in circumstances inhibiting detectable adsorption of the antigen. Antigen adsorption was inhibited by proportionate excesses of protein not involved in the antigen-antibody reaction. Evidence for the adsorption of measurable amounts of antigen-antibody complexes in association with agglutination of the latex particles for conditions suppressing antigen adsorption was not obtained.
Biological Activities of Aggregated γ-Globulin
Authors: Ishizaka Kimishige , Ishizaka Teruko , Banovitz Jay
Published: January 2023

The minimum size of nonspecifically aggregated rabbit γ-globulin having skin reactivity and C′-fixing properties was studied. The results indicated that 9.2 to 9.6 S component in normal and biphenylene azo-RGG preparations had both activities, whereas nonaggregated γ-globulin of 6.6 S did not have either activity. The C′-fixing activity of the 9.6 S component was less than that associated with aggregates larger than this component. The smallest aggregate of piece III of the γ-globulin having both activities was comparable to or slightly smaller than 7 S γ-globulin. The results suggested that induction of these biologic activities was associated with dimerization of γ-globulin or piece III.
Antibodies Produced by Guinea Pigs Infected with Foot-and-Mouth Disease Virus
Authors: Cowan K. M. , Trautman R.
Published: January 2023

Guinea pigs infected with foot-and-mouth disease virus produced 19 S-γ1 antibodies that were detected by agar gel precipitin tests at the 4th day following inoculation. This antibody type persisted for a short time and was not detectable by precipitin tests after 15 days. The 19 S antibody neutralized virus, adsorbed firmly to diethylaminoethyl (DEAE) cellulose, passed rapidly through Sephadex G-200, but did not fix complement. About 10 days following infection, 7 S-γ2 antibody was demonstrable and persisted. It neutralized virus, precipitated in agar gel tests, fixed complement, bound weakly to DEAE-cellulose, and passed more slowly through Sephadex G-200 than 19 S antibody.

The ultracentrifugal separation of 7 S and 19 S classes of antibody was done in an angle rotor using a KBr-NaNO3 gradient. This procedure made it possible to run as much as 1.0 ml of undiluted serum/tube.
Use of Mouse Serum in Identification and Serologic Classification of Rickettsia Akari and Rickettsia Australis
Authors: Pickens E. G. , Bell E. J. , Lackman D. B. , Burgdorfer W.
Published: January 2023

Following infection with R. akari or R. australis, Swiss mice developed species-specific antibody which, unlike guinea pig convalescent sera, did not fix complement with the soluble, cross-reacting antigen of heterologous rickettsias of the spotted fever group. The EM rickettsia, a newly described agent recovered from ticks from eastern Montana, also produced highly specific CF antibody in mice. Cross-reactions were not observed between the EM agent and any of the members of the spotted fever group tested. Species-specific antibody was produced in mice by other agents of the group, i.e., R. rickettsi, R. conori and R. siberica. Yet, because of the extremely large dose of these rickettsias required and the erratic results obtained, these studies are regarded as preliminary only. Toxin-neutralizing, but not CF, antibody could be detected in sera of mice inoculated with relatively low doses of live or inactivated R. rickettsi.
Relation of Antibody Titer to Persistence of Antigen
Authors: Richter Maxwell , Zimmerman Sarah , Haurowitz Felix
Published: January 2023

The antibody titer in the serum of rabbits injected with bovine serum albumin or ovalbumin was determined over a period of 1 to 2 years. The decrease in antibody titer, several months after the last injection, parallels in general the decrease in the amount of persisting antigen.
The Radioisotope Precipitation Test for Study of Q Fever Antibodies in Human and Animal Sera
Authors: Tabert Gayle G. , Lackman David B.
Published: January 2023

The radioisotope precipitation test (RIPT) for measuring Q fever antigen-antibody reactions was used to examine various groups of human and animal sera. The specificity of the RIPT was determined by testing sera of Alaskan natives from Q fever-free areas, prevaccination sera from Q fever vaccination experiments in man and four sera from an egg-sensitive person. Only two of the sera from persons in the Alaskan group, one of whom had a strongly positive skin test, were RIPT positive. Only four of the 173 prevaccination sera were RIPT positive and no reaction was noted with the sera from an eggsensitive person. Results on the vaccine groups and those groups occupationally exposed demonstrated that the sensitivity of the RIPT is much greater in man than the complement fixation or capillary agglutination tests. The finding that there are so few cases of Q fever among exposed persons may be partly explained by the high percentage already having antibody detectable by the RIPT.

The test was further applied to cattle, guinea pig and mouse sera but the increased sensitivity observed with human sera was not present with animal sera although it could be used for testing them.
Natural Resistance of Germ-Free Mice and Colostrum-Deprived Piglets to Group A Streptococci
Authors: Stollerman Gene H. , Ekstedt Richard D. , Cohen Irun R.
Published: January 2023

A study was made of the resistance of germfree and conventional mice to direct challenge with Group A streptococci of graded virulence. Germ-free and conventional mice were highly, and equally, resistant to nonencapsulated strains of Group A streptococci lacking M protein. Greater susceptibility of germ-free mice could be demonstrated only with Group A strains containing both M protein and capsules. This difference could be abolished by a globulin fraction of normal mouse serum but not by normal mouse γ-globulin or by homologous type-specific antiserum.

In vitro studies with bloods of germ-free mice and colostrum-deprived piglets provided additional evidence that phagocytosis of avirulent Group A streptococci required little, if any, specific antibodies. Bloods from these animals phagocytized and destroyed streptococci as readily as did their conventional counterparts. Thermolabile opsonins against avirulent Group A streptococci were readily demonstrable in the blood of colostrum-deprived piglets, whose serum contains only minute amounts of γ-globulin. Absorption of the sera of these animals at low temperatures with large amounts of homologous streptococci resulted in some decrease in opsonization, but considerable residual phagocytosis was apparent. The leukocytes of colostrum-deprived, and of colostrum-fed, animals were equally capable of phagocytizing and destroying Group A streptococci in the presence or absence of specific opsonins.
The Role of Collagen in the Induction of Flea Bite Hypersensitivity
Authors: Michaeli D. , Benjamini E. , de Buren F. P. , Larrivee D. H. , Feingold B. F.
Published: January 2023

Salt-soluble and acid-soluble collagens were isolated from guinea pig skin following bites of fleas, and were shown to possess the capacity to induce flea bite hypersensitivity upon injection into recipient guinea pigs. Less salt-soluble collagen was required for induction of sensitivity than was acid-soluble collagen. Flea hapten associated with salt-soluble collagen did not hinder the latter's in vitro reconstitution. The in vitro reconstituted product served as a better carrier for the hapten than the nonreconstituted form.
Antigenic Differentiation of Polioviruses by Absorbed Hyperimmune Sera
Authors: Eylan E. , Fogel A. , Dukler R. , Rannon L.
Published: January 2023

The antibody absorption method for the preparation of highly specific antisera against Type I vaccine and virulent strains has been applied in an epidemiologic study.

A total of 119 Type I polioviruses isolated in Israel during a 3-year period were tested for their antigenic relationship to LSc-2ab virus. All isolates from the prevaccination period were different from the vaccine, enabling identification of strains from these groups by antigenic marker. Strains excreted by vaccinated children early after feeding (1 week) were also clearly classified, in this case as vaccine-like. In viruses excreted by vaccinated children in later periods after feeding, antigenic change, observed also by other authors, was confirmed by the antibody absorption method. The greater sensitivity of this method is emphasized.
Studies of Herpes Simplex Virus by the Gel Diffusion Technique
Authors: Tokumaru Tadasu
Published: January 2023

Herpes simplex virus obtained from infected rabbit kidney cell tissue cultures contained high titers of precipitating antigens, which proved suitable for use in a diagnostic test for precipitins in primary and recurrent herpes infections. The use of this test in the screening of the sera of 671 persons has provided valuable information regarding human precipitins, particularly with respect to the existence of wide variations in the antigen-antibody systems involved. The precipitin tests used in these studies should be of value for the detection of infections in patients with passively acquired circulating antibodies. A high incidence of primary and recurrent herpes simplex virus infections of the upper respiratory tract were observed in the present study.
Kinetics of the Bactericidal Action of Normal Serum on Gram-Negative Bacteria
Authors: Davis Starkey D. , Wedgwood Ralph J.
Published: January 2023

The kinetics of the bactericidal action of serum upon Gram-negative organisms was studied with a viable plate count assay. This was found to be a first-order reaction consisting of a definite lag period followed by killing at a constant logarithmic rate. The Q10 of this reaction was about 2. The lag phase probably represents the time required for the formation of intermediate products. The bactericidal reaction was markedly affected by bacterial growth phase. Bacteria in the resting state were resistant to serum, and bacteria in the rapid growth phase were sensitive to serum. In addition, the culture temperature was found to markedly affect the subsequent bactericidal reaction. Bacteria cultured at 37°C were less sensitive to killing at 27°C than those originally cultured at 27°C.
Chromatographic Purification of Tetramethylrhodamine-Immune Globulin Conjugates and Their Use in the Cellular Localization of Rabbit γ-Globulin Polypeptide Chains
Authors: Cebra John J. , Goldstein Gerald
Published: January 2023

Rabbit spleen imprints were double stained with pairs of fluorescein and tetramethylrhodamine conjugates derived from the following goat reagent globulins: anti-Fragment III (anti-γG heavy chain), anti-light chain and anti-Fragment I. To obtain specific staining of the component chains of rabbit immunoglobulin it was necessary to develop procedures for the chromatographic fractionation of tetramethylrhodamine-immune globulin conjugate to supplement existing procedures for the preparation of specific staining fluorescein-conjugates. Bright specific staining could be obtained using the subfractionated conjugates at concentrations as low as 32 to 43 µg of antibody/ml. A series of simple Kodak Wratten filters, K2, 23A and 57A, were used to differentiate between singly and doubly stained fluorescent cells. These filters permitted detection and proper classification of fluorescent cells even when the optimal ratio of reagent concentrations was varied over a 16-fold range. Staining with anti-light chain and anti-III, conjugated with contrasting fluorochromes, resulted in double labeling of 63 to 78% of all fluorescent cells. These cells presumably contained all component chains of γG-globulin. A distinct minority of the fluorescent cells, 21 to 34% of the population, stained only with the anti-light chain reagent. A similar class of cells, comprising 14 to 26% of all fluorescent cells, stained only with the anti-I reagent when it was used as a counterstain with anti-III reagent. Anti-light chain reagents detected more immunoglobulin-containing cells than did anti-I reagents. Double staining with anti-I and anti-light chain, conjugated with contrasting fluorochromes, yielded 63 to 85% doubly stained cells, 15 to 32% of cells stained with the anti-light chain alone, and 0 to 10% of cells stained with anti-I alone.
The Stability of Messenger Ribonucleic Acid in Antibody Synthesis
Authors: Lazda Velta , Starr Jason L.
Published: January 2023

Spleen fragments obtained from hyperimmune rabbits 3 days after secondary immunization continued to form antibody in vitro for at least 18 hr after preincubation with actinomycin D. Total cellular ribonucleic acid (RNA) synthesis was decreased by about 75% and was limited to nucleotide incorporation into soluble RNA. Antibody synthesis in actinomycin-treated cells was markedly diminished after 24 hr, at which time RNA synthesis was beginning to increase. The results are consistent with the hypothesis of a relatively stable messenger in mammalian protein synthesis, in contrast with the unstable messenger RNA observed in bacterial cells. A revision in the concept of nonspecific ribosomes may be necessary for animal cells.
Mercaptoethanol-Sensitive Antibody Production in Germ-Free Mice and Guinea Pigs
Authors: Sell Stewart
Published: January 2023

Germ-free and nongerm-free guinea pigs and mice were injected intraperitoneally or in the footpads with soluble I131 bovine γ-globulin (BγG) or BγG precipitated with alum. The germ-free guinea pigs responded to these injections in a manner similar to the nongerm-free guinea pigs. Germ-free mice injected intraperitoneally also responded in the same manner as nongerm-free mice, but germ-free mice injected in the footpads produced only a delayed 19 S antibody response while nongerm-free mice injected in the footpads demonstrated a prompt antibody response with a 19 S-to-7 S conversion. It is suggested that this difference in antibody response in germ-free mice after different routes of injection indicates that the number of “potentially competent” cells available to a given antigen may determine the type of antibody produced.
Roles of the Liver and Appendix in the Serum Hemolysin Response in Rabbits
Authors: Draper Laurence R. , Sussdorf Dieter H.
Published: January 2023

The roles of the liver and appendix in the serum hemolysin response in rabbits immunized with one or more injections of sheep erythrocyte stromata were studied. The effects of extirpation of these organs were observed both in the presence and absence of the spleen. Antibody titers were determined photometrically in 50% hemolysin units, and the serum hemolysin response was analyzed with respect to the lengths of the latent period and rise to peak titer, day and magnitude of peak titer, and rates of hemolysin rise and decline.

Using this approach, no antibody contribution by the appendix or liver to the serum hemolysin response was detected. These organs apparently were inactive even after repeated injections of antigen which overrode the depressive effect of splenectomy by inducing increased nonsplenic activity.

Changes in the hemolysin response observed following immunization via the portal vein and in partially hepatectomized animals injected via the ear vein could be attributed to the liver's capacity to remove significant amounts of antigen from the circulation. Increased disappearance rates of passively transferred hemolytic antiserum in partially hepatectomized and liver-irradiated rabbits suggest that the regenerating or recovering liver catabolizes antibody faster.
The Effectiveness of Acriflavine and Ineffectiveness of Proflavine as Immunosuppressant Agents
Authors: Samuelson Joel S. , Stewart P. Brian , Kraft Sumner C. , Farr Richard S.
Published: January 2023

Acriflavine hydrochloride N.F. is an immunosuppressant agent containing approximately 80% acriflavine and 20% proflavine. The structures of these two diaminoacridines differ only with respect to a methyl group at the 10-position of the acridine nucleus. Antibody production to BSA in rabbits was suppressed by acriflavine but not by comparable or higher doses of proflavine. The degree of immunosuppression was related to the amount and duration of acriflavine administration. One course of acriflavine treatment during the first exposure to BSA was sufficient to induce a degree of immune paralysis to a subsequent dose of BSA. Acriflavine administered via the oral route was absorbed poorly and resulted in a high mortality.
Influence of Antigen Dosage on Kinetics of Hemagglutinating Antibody Production
Authors: Albright J. F. , Evans T. W.
Published: January 2023

We have studied the effects of antigen dosage upon the kinetics of hemagglutinating antibody production by both primed and nonprimed spleen cells. For these studies the method of in vivo culture of spleen cells was employed, thus avoiding many of the complicating factors which have attended previous studies in intact animals. A broad range in both competent spleen cells and antigen dosage (rat erythrocytes) was examined. Antibody response was characterized by regions of antigen insufficiency, optimum antigen and antigen excess. Detailed response profiles indicated that the slopes of the logarithmic phase of antibody production were independent of antigen dosage and characterized by antibody-doubling times of about 8 hr, a figure which is consistent with the known cell cycle time of 9 hr of blast cells involved in antibody response. Thus, marked variation in either the cell cycle time or the amount of antibody produced per functional cell per cell cycle during a given response appeared not to occur with variation in antigenic stimulus. The variable factors which seem most likely to be affected by antigen dosage were concluded to be: a) variation in the number of activated progenitor cells, b) variation in the number of functional progeny generated by progenitor cells, and c) variation in the initial, but subsequently constant, amount of antibody produced per functional cell per cell cycle. It was further concluded that variation in antigen dosage is not simply all-or-none activation of progenitor cells.
Experimental Infection of Parainfluenza Virus Type 5 in Mice, Hamsters and Monkeys
Authors: Chang P. W. , Hsiung G. D.
Published: January 2023

Experimental infection of DA virus in mice, hamsters and monkeys has been studied and the pathogenesis of this virus in these animal species has been analyzed. Inapparent infections have been demonstrated in all three animal species. Infection of hamsters and monkeys with DA virus induced high levels of antibody. Such animals were resistant to further infection by the virus. In mice, low level or frequently no detectable antibody was found following infection, and these animals could be reinfected.

DA virus was isolated from a number of organs from infected animals. The appearance of the virus in different organs followed a definite pattern, depending on the route of inoculation. Viremia was detected infrequently following intranasal inoculation, and viremia may have served as a means in the dissemination of the virus from the primary site of infection, i.e., lung and trachea, to the liver, kidneys and brain. Minor histopathologic changes were found in the lung tissue of infected mice, but none was found in the trachea, liver or kidney. Virus was recovered in the urine of mice following intraperitoneal inoculation and in hamsters following intranasal inoculation. DA virus persisted in kidneys of rhesus monkeys, even in the presence of humoral antibodies. Persisting virus could not be detected in the kidneys by the conventional isolation methods, but virus could be recovered by growing the kidney cells in monolayers on glass and maintaining these cultures for a period of 1 to 2 months.
Immunologic Reactivity in Neonatally Thymectomized Rats Receiving Competent Lymphoid Cells during Immunization
Authors: Isaković Katarina , Waksman Byron H. , Wennersten Christine
Published: January 2023

Adult Lewis rats, injected with bovine serum albumin and complete Freund's adjuvant in a hind footpad, develop intense delayed skin reactivity and both hemagglutinating and precipitating antibody, accompanied by skin reactivity of the Arthus type. Animals thymectomized within 24 hr of birth fail to develop these responses when challenged at 8 to 10 weeks of age. If, however, they receive intravenous infusions of normal isologous lymph node cells at the time of challenge, they react immunologically in a manner closely similar to controls. The intensity and time course of each immune response are correlated with the dose and the time of administration of cells, respectively. The blood lymphocytes and the small lymphocyte population of the spleen and lymph nodes return towards normal levels in transfused animals. The experiment fails in noninbred Sprague-Dawley rats, which develop homologous disease when injected with lymph node cells. Adult thymus cells, even in very large numbers, are ineffective in restoring reactivity in Lewis rats, whether given at the time of challenge or 4 to 8 weeks earlier.
Passive Uveal Anaphylaxis in Guinea Pigs
Authors: Lanzieri Mario
Published: January 2023

Passive anaphylactic reactions were obtained in albino guinea pigs by injection of antibody into the vitreous body or into the anterior chamber of the eye.

When the antibody is injected into both the vitreous body and the anterior chamber, only the anterior part of the uvea participates in the reaction. Edema of the tissues, changes in the permeability of the capillaries (especially small veins), presence of polymorphonuclear leukocytes in large numbers in the anterior chamber and around the ciliary body are observed.

The posterior uvea does not seem to participate in these anaphylactic reactions.
Antigens in Immunity
Authors: Nossal G. J. V. , Ada G. L. , Austin Caroline M.
Published: January 2023

When newborn rats were injected intraperitoneally with 1 µg or more of Salmonella adelaide flagellin on the day of birth and thereafter twice per week until adult life, they developed immunologic tolerance to this powerful bacterial antigen. Similarly, a single injection into newborn rats of a larger amount (100 µg) of flagellin induced complete tolerance, though this was not as long-lasting as the tolerant state following multiple injections.

Rats rendered tolerant to monomeric flagellin also showed a marked degree of partial tolerance to the particulate antigens, polymerized flagellin and intact flagella.

When these particulate antigens were injected repeatedly during the newborn period, it was shown that multiple doses of polymerized flagellin produced nearly complete tolerance; on a weight basis, polymerized flagellin was 10 times less efficient than monomeric flagellin. Only a slight degree of tolerance could be produced by multiple injections of intact flagella.

The chief value of the study was twofold: First, it showed that convincing tolerance can be induced toward highly “foreign” microbial antigens. Second, elucidation of the minimum amount of antigen capable of giving complete tolerance has set the stage for a meaningful study of the cellular distribution and persistence of antigen molecules responsible for tolerance induction, using radioactively-labeled flagellin and autoradiography.
Complement Fixation Studies with a Varicella-Zoster Antigen
Authors: Gold Eli , Godek Geraldine
Published: January 2023

Discussion and Summary

A satisfactory complement-fixing varicella or zoster antigen can be prepared from tissue culture cells infected with these agents. The two agents are antigenically indistinguishable and either antigen may be used to measure the antibody response to either infection. The studies with the complement fixation test are consistent with the results of other investigators (3, 4) and provide the basis for the statement that varicella and herpes zoster viruses are identical.

The apparently more rapid development of antibody following zoster in comparison with varicella may represent a secondary or booster response, or may simply indicate that dating the onset by the appearance of the rash may be erroneous. Most patients with zoster have a history of paresthesia or some form of discomfort in the area in which the rash later appears. Perhaps this symptom should be considered the time of the onset of the disease. The failure of antibody to persist for a greater duration following zoster than after varicella makes it even more difficult to support the concept of a secondary immune response.

The fall in antibody levels within 4 years or less following infection was not accompanied by any apparent change in susceptibility to infection; no second attacks of varicella or zoster were observed. Re-exposure of children with a history of chickenpox to that infection did not produce a rise in antibody levels.

Complement-fixing antibodies to the agents of zoster or varicella may fall more quickly than neutralizing or other types of antibody (as occurs with the viruses of mumps, measles, poliomyelitis, and others), or levels of antibody not measurable by present techniques may be sufficient to protect from infection. A sensitive test for neutralizing antibody is needed to resolve this point. It is clear from the studies presented, however, that the CF test may be used to measure the antibody response following zoster or varicella, but it is of little value for determining susceptibility to infection.
Synthesis of Components of Sendai Virus in the Presence of Proflavine
Authors: Azadova N. B. , Uryvayev L. V. , Zhdanov V. M.
Published: January 2023

Proflavine sharply alters the processes of transport of S-antigen of Sendai virus from the nucleolus to the cytoplasm, but does not essentially decrease its synthesis. The decrease in synthesis of the V-antigen is proportionately more marked. Retention of S-antigen in the nucleolus leads to the formation of incomplete virus and to a reduction of release of infectious virus from the cell. Interaction of proflavine with the viral RNA does not essentially alter the messenger mechanism responsible for synthesis of protein components of the virus.