bluuebunny/crossref_metadata_2025 huggingface dataset:
The Effects of Proteolytic Enzymes on Agglutination by Bacterial Antibodies of Lipopolysaccharide Modified Erythrocytes
Authors: Neter E. , Cohen E. , Westphal O. , Lüderitz O.
Published: January 2023

The effects of various proteolytic enzymes on agglutination of antigen modified erythrocytes from both warm and cold blooded animal species were investigated, and the following results were obtained.

Red blood cells from alligator, axolotl and caiman after treatment with enterobacterial antigens (crude extracts and highly purified lipopolysaccharides) are not agglutinated by homologous bacterial antiserum of rabbit and human origin, in contrast to sheep and human erythrocytes. Specific hemagglutination of lipopolysaccharide modified erythrocytes from these cold blooded animal species is made possible by treatment of the red blood cells with trypsin, chymotrypsin, protease (of pancreatic origin), ficin and papain. Identical results are obtained when enzyme treatment precedes or follows antigen modification. The agglutination enhancing effect of trypsin is abolished by soy bean trypsin inhibitor. Proteinase (of fungal origin), peptidase (from hog intestinal mucosa) and human plasmin are ineffective in this system. Trypsin, chymotrypsin, protease, ficin and papain also enhance agglutination of alligator erythrocytes by alligator red blood cell antiserum, although to a substantially lesser degree than agglutination of lipopolysaccharide modified erythrocytes by homologous bacterial antibodies. Alligator, axolotl and caiman erythrocytes after treatment with the proteolytic enzymes are not agglutinated by either homologous or heterologous bacterial antisera or normal rabbit serum in the dilutions used, indicating that a Thomsen-Friedenreich type of reaction is not encountered. Lipopolysaccharide modified human erythrocytes of blood group A or B after treatment with the proteolytic enzymes are agglutinated to a slightly greater degree by both homologous bacterial and blood group antisera than are red blood cells not treated with enzymes. Lipopolysaccharide modified sheep erythrocytes after treatment with trypsin, chymotrypsin, protease, ficin and papain are agglutinated to the same extent by homologous bacterial antiserum as are erythrocytes not treated with enzymes. Enzyme treatment, however, enhances agglutination by sheep cell antiserum (amboceptor). It is postulated that erythrocytes from certain cold blooded animal species, in contrast to those from sheep and man, contain on their surface in the area of the receptors of the lipopolysaccharide a protein or protein-like material which interferes with the spatial orientation of bacterial antibodies and subsequent hemagglutination, and that certain proteolytic enzymes remove or alter this inhibitor making hemagglutination possible. The bearing of the results obtained in this study on the nature of incomplete antibodies and of the agglutination by bacterial antibodies of erythrocytes modified by homologous bacterial antigen is discussed.
Immunity against a Transplantable Ascites Tumor of Spontaneous Origin in an Inbred Rat Strain
Authors: Goldner Herman , Bogden Arthur E. , Aptekman Paul M.
Published: January 2023

Rats of the inbred P.A. strain have been immunized against the 9A ascites tumor, which originated spontaneously in an animal of the same strain. This iso-immunity appears to be of the humoral type. The humoral antibody detectable in the serum of immunized rats has a cytotoxic action on the 9A ascites tumor and is capable of inactivating tumor cells both in vitro and in vivo. Therapeutic and prophylactic administration of immune serum afford complete protection of normal rats from lethal intraperitoneal tumor implants or extends their survival time. The ratio of the number of tumor cells to the volume of immune serum is cn'tical in such therapy.
Studies of Blood Group Antibodies
Authors: Abelson Neva M. , Rawson Arnold J.
Published: January 2023

A simple method is described for effecting separations of Rh saline agglutinins from other Rh antibodies. The method also can be used to separate Coombs-active and albumin-active antibodies from inert γ-globulins and from the bulk of the isohemagglutinins anti-A and anti-B.
Localization of I131-Labeled Antirat Lymph Node and Kidney Rabbit Antibodies in the Rat
Authors: Moore Richard
Published: January 2023

Summary and Conclusions

A homogenate of rat lymph node was injected into rabbits to stimulate production of antirat lymph node antibodies. The rabbit globulin was radioiodinated and adsorbed on and eluted from rat lymph node homogenate. The product was tested in vitro by precipitation tests against various organ homogenates, and radioassays showed the presence of labeled antilymph node antibodies.

The data from the rats injected subcutaneously and serially sacrificed showed the presence of labeled antibodies which localized in lymph nodes more than did labeled antikidney antibodies or labeled globulin from normal serum. Labeled antilymph node antibodies localized in spleen and mesenteric lymph nodes more than did labeled antikidney antibodies or labeled globulin from normal serum. In this experiment, antibodies prepared against lymphatic tissue differed qualitatively from other proteins in their lymphatic tissue localization.

An alternative method of preparation was employed in which antibody separation was followed, rather than preceded, by isotope labeling. Antibodies so labeled were injected intravenously and subcutaneously for an in vivo test similar to the previous experiment. Intravenous injection resulted in maximum antibody localization in spleen. Subcutaneous injection resulted in maximum antibody localization (up to 25% of the significant dose) at the injection site and the lymph nodes which drained it.

As is well known, lymphatic tissue is scattered throughout the body with some portions lying well below the skin and not easily found. A technique for tracing the tissues of this system would provide information to immunologists tracing antigen-antibody combinations, and, conceivably, to surgeons performing radical lymph node dissections. The data presented are consistent with the hypothesis that lymph nodetissue specific antibody had been prepared and labeled and that the antibody, when injected, traced the lymphatic flow of the animal.
Quantitative Studies on the Antigen-Antibody Reaction by the Monolayer Technique
Authors: Ogata Tomio , Tachibana Taro , Suzuki Kan , Suzuki Teruko
Published: January 2023

The monolayer technique in the observation of the antigen-antibody reaction is applicable to desorption experiments irrespective of the valency of the antigenic substance. It offers a new method of measurement of the strength of combination between haptens and antibodies. This strength can be expressed as relative desorption strength.

The values of relative desorption strength significantly depend upon the number of atoxyl groups per molecule and higher values are obtained with the haptenic compounds containing greater number of atoxyl groups. However, differences exist in the desorption strengths of haptens with the same valence. Among the substances with two atoxyl groups, the atoxyl-azoamino acids, which are closer in chemical structure to the immunizing antigen, exhibit higher values than any other compounds. In addition they give no precipitate with the antiserum in contrast to other haptens with two atoxyl groups.

It seems reasonable to assume that the haptens bearing two atoxyl groups in meta position constitute one single determinant group. In that sense it may be considered as univalent contrary to the current concept that a haptenic substance with two atoxyl groups is bivalent under any circumstances.
The Properdin System and Immunity
Authors: Lepow Irwin H. , Pillemer Louis , Schoenberg Melvin D. , Todd Earl W. , Wedgwood Ralph J.
Published: January 2023

Human properdin, purified 2500-fold over serum by adsorption to and elution from Zymosan, was found to be heterogeneous by biochemical, immunological, electrophoretic, and ultracentrifugal criteria. Electrophoretically, properdin activity was associated with γ1-globulins; ultracentrifugally, primarily with a component having a sedimentation constant of 19 S at infinite dilution. Purified properdin was antigenically heterogeneous and was contaminated with a variety of normal human serum constituents. In all cases, including phosphatase, lysozyme and Zymosan agglutinin, these could be distinguished from properdin itself.
Immunochemistry of Synthetic Polypeptides and Polypeptidyl Proteins
Authors: Stahmann Mark A. , Lapresle Claude , Buchanan-Davidson Dorothy J. , Grabar Pierre
Published: January 2023

Bovine albumin, modified by the addition of polypeptides of glutamic acid, lysine, leucine, or phenylalanine was found to be antigenic in rabbits. Evidence is presented to show that antibodies specific for the added peptide modification and for the carrier protein were formed. In addition a third type of antibody may have been formed to the polyleucyl and polyphenylalanyl bovine albumins, which requires the added peptide and at least a part of the carrier protein. Rabbit albumin modified in the same way was also antigenic in rabbits. Antibodies were formed to the polyleucyl and polyphenylalanyl rabbit albumins which were specific for the modification and which also required the added peptide and at least part of the carrier protein. Antisera to the polypeptidyl proteins cross reacted with several purified protein preparations with which antisera to the carrier proteins did not react.
Studies of Aggregated γ-Globulin
Authors: Christian Charles L.
Published: January 2023

Injection of aggregated human γ-globulin into the skin of guinea pig, rat and man produced inflammatory changes. Histologically, the injection sites of guinea pigs and rats demonstrated marked polymorphonuclear and mononuclear infiltrations that were maximal at about 24 hr.

Intravenous administration of aggregated human γ-globulin, which is strikingly anticomplementary in vitro, rendered the sera of recipient animals deficient in hemolytically active C′ for several hours.
Studies on Variation in Coxsackie A-9 Virus
Authors: Hsiung G. D.
Published: January 2023

After 6 to 14 serial passages of Coxsackie A-9 virus in x-irradiated patas cultures, a strain was obtained which a) multiplied in patas cultures, b) was reduced in virulence for new born mice, and c) produced only small plaques (5 mm at 7 days) on both rhesus and patas monolayers.

A single passage of the adapted virus either in mice or in rhesus cultures (both in fluid medium and under agar), shifted the virus population toward the original large plaque type and tissue culture host range.

Reversion to the large plaque type was observed after 4–8 hr growth (single growth cycle) when the adapted virus at limiting dilutions was transferred to rhesus cultures. However, if harvests were made after 24–48 hr a mixed plaque size population was obtained. This was attributed to a slower growth rate in the adapted virus.
Propagation of Certain Arthropod-Borne Viruses in Avian and Primate Cell Cultures
Authors: Henderson J. R. , Taylor R. M.
Published: January 2023

Multiplication in tissue culture of 21 arthropod-borne viruses representative of serogroups A, B and C was manifested by a consistency of plaque formation under an agar overlay. Cytopathic reactions in infected fluid cultures were not always apparent even though virus multiplication was detected. In comparative virus titrations in tissue cultures and infant mice, the plaque assay was found to be the most sensitive system. In general, group A viruses were found to multiply in both avian (chick embryo and Pekin duck kidney or embryo) and the primate (rhesus monkey kidney) cell systems with few exceptions. Group B viruses could be propagated in Pekin duck kidney or Pekin duck embryo cultures, but not in chick embryo or rhesus monkey kidney (except Ilheus), and the group C viruses produced plaques only in rhesus monkey kidney cell cultures.
Studies on Organ Specificity
Authors: Rose Noel R. , Metzgar Richard S. , Isaacs Eli
Published: January 2023

The sera of rabbits immunized with hog thyroid crude saline extract or hog thyroglobulin were studied by gel-diffusion precipitation techniques. Several precipitation lines were discernible by the double diffusion techniques of agarprecipitation in tubes when hog thyroid antisera were tested with hog thyroid extract. The individual precipitation bands were identified by absorption experiments. The antisera were shown to contain antibodies that react only with antigen(s) in hog thyroid extract as well as antibodies that cross-react with antigen(s) in thyroid extracts from other species. In addition, some of the antisera contained antibodies that react with extracts of other hog organs. The nature of the reactions of hog thyroid antisera with hog thyroid extract and thyroid extracts from other species was demonstrated by the Ouchterlony technique of gel precipitation. One of the two lines produced by the reaction of a hog thyroid antiserum (after species-specific antibodies had been absorbed) with hog thyroid extract gave a reaction of partial identity with a single precipitation band produced by the same antiserum with the thyroid extracts of certain other species. Hog thyroid antisera (species-specific antibodies absorbed) gave a single precipitation are with hog thyroid extract and thyroid extracts of other species subjected to electrophoresis. The location of the precipitation are was the same for both the homologous and cross-reactions. The organ-specific thyroid antigens have the same electrophoretic mobility as α-globulins of human serum.
A Possible Correlation between the Minimum Sensitizing Dose of Antibody in Passive Anaphylaxis and the Molecular Weight of the Antigen
Authors: Pruzansky Jacob J. , Feinberg Samuel M. , Harris Mildred , Wojcik Norma
Published: January 2023

Rabbit and guinea pig antisera to four antigens were compared as to their ability to sensitize guinea pigs passively. The minimum sensitizing dose of each set of sera appeared to be related to the molecular weight of the corresponding antigen. Nonprecipitating antisera were indistinguishable quantitatively from precipitating antisera.
Changes in Immunity and Antitoxin Level Immediately after Secondary Stimulus with Tetanus Toxoid in Rabbits
Authors: Ipsen Johannes
Published: January 2023

Injections of 5 Lf of fluid tetanus toxoid were given to 79 rabbits. Four weeks later circulating antitoxin was measurable in all but six of the animals. At that time booster injections of 5 Lf fluid tetanus toxoid were given to experimental groups whereas other groups were maintained nonstimulated. At various times from 3 hr to 8 days after the stimulus blood samples were taken from all rabbits, after which an intramuscular injection of 1000 lethal rabbit doses of tetanus was given. Prolongation in survival time was shown to start immediately after the stimulus whereas antitoxin level showed no significant rise in the first 2 days after stimulus. It is inferred that the lag in antitoxin response after tetanus booster dose not reflect a lack of response in terms of total body immunity.
Studies on the Immunologic Mechanism of Anaphylaxis
Authors: Ovary Zoltan , Karush Fred
Published: January 2023

The fractions obtained from purified rabbit antihapten antibody by papain digestion, which contain the antibody combining site (ADDS or ADS-I), were unable to sensitize the skin of the guinea pig for PCA or RPCA reactions. These fractions, however, when mixed with the intact antibody specifically inhibited the latter from producing PCA reactions showing that they can combine in vivo with the antigen. From a kinetic study it was concluded that these fractions diffuse away faster from the injection site than does the intact antibody. These fractions showed a precipitation line in agar double diffusion with horse-anti-rabbit γ-globulin. The fraction without antibody combining site (ADDP) can produce good RPCA reactions. It was concluded that the failure of ADDS and ADS-I to produce PCA or RPCA reactions is due to their inability to be “fixed” to the guinea pig tissues and that this “fixation” site is present on ADDP.
Studies on the Combining Sites of the Protein Antigen Silk Fibroin
Authors: Cebra John J.
Published: January 2023

The following peptides have been isolated on a preparative scale (100–500 mg) from soluble chymotryptic hydrolysates of fibroin: Gly(Gly, Ala)Tyr; Gly(Gly, Val)Tyr; Gly(Gly3,Ala3)-Tyr; Gly(Gly3,Ala2, Val(Tyr; Ala(Gly2, Val, Asp)Tyr; and a mixture of dodecapeptides of the form [Gly(Gly3,Ala3)Tyr][Gly(Gly, Val)Tyr], [Gly(Gly3,Ala2, Val)Tyr] [Gly(Gly,Ala)Tyr] and so forth.
Restoration of Serum Bactericidal Activity and Prevention of its Loss in X-Irradiated Mice
Authors: Kornfeld Lottie , Miller C. Phillip
Published: January 2023

The loss of normal serum bactericidal activity of CF-1 mice for a strain of Escherichia coli which occurs 12 hr after total-body x-irradiation (600 r) was prevented if yeast autolysate, yeast ribonucleic acid, homogenates of normal mouse spleens or bacterial endotoxin were injected ½ hr before or ½ hr after exposure to x-rays. Injection 18 hr before irradiation was without effect. Injection of any of these substances during the postirradiation period, when bactericidal activity was absent, resulted in the appearance of bactericidins 18 hr after treatment. Spleens removed from donor mice 1 or more hours after exposure to 600 r failed to restore bactericidins in irradiated recipients.
Biological Activity of Aggregated γ-Globulin
Authors: Ishizaka Kimishige , Ishizaka Teruko , Sugahara Takao
Published: January 2023

A single injection of aggregated human and rabbit γ-globulin, obtained by coupling with bis-diazotized benzidine, induced Arthus-like reactions in normal guinea pigs. The aggregated γ-globulin and soluble BSA-anti-BSA complexes were comparable on a weight basis with respect to their biologic activity. The activity of aggregated γ-globulin was dependent on the animal species from which the γ-globulin was derived. Neither aggregated bovine nor chicken γ-globulin showed activity. Horse γ-globulin induced the skin reaction upon aggregation, although it was less active than the aggregated human and rabbit γ-globulins. Aggregated human γ-globulin prepared by heating and by coupling with bis-diazotized benzidine induced erythema-wheal reactions in humans.
Agglutinin Response, a Function of Cell Number
Authors: Perkins Eugene H. , Robinson M. A. , Makinodan T.
Published: January 2023

Our in vivo culture studies revealed that, independent of a) the strain of mice; b) the type of particulate antigen; c) the time of bleeding after cell transfer and antigen injection; and d) whether animals were undergoing a primary or secondary antibody response, a linear log2 antibody titer log2 nucleated cell relation was obtained with spleen and peritoneal exudate cells. Furthermore, the slopes of the resulting regression lines for the primary and secondary responses did not differ significantly, but the antibody-forming activity per given number of nucleated cells was significantly greater in the secondary antibody response than in the primary antibody response. The optimum dose of RBC for a maximum sensitization was 1 ml of 1%, and no difference between intravenous and intraperitoneal routes of RBC injection was noted.
Studies on Hemagglutinins in California Wild Flower Seeds
Authors: Brilliantine Laura , Allen Nancy K.
Published: January 2023

Saline extracts of 215 varieties of California wild flower seeds belonging to 42 families were tested for the presence of hemagglutinins specific for human blood factors. No specificity for A, B, O, M, N or Rh antigens was found.

Of the 215 extracts studied, 31 showed hemolytic activity and 158 were inactive. Twenty-six were nonspecific; of these, 7 were nonspecific in saline and 19, although nonreactive in saline, were nonspecific after the addition of polyvinyl pyrrolidine.
A Mortality Enhancing Extract Isolated from Listeria Monocytogenes
Authors: Silverman Sidney J. , Elwell Lynn , Kautter Donald A.
Published: January 2023

The injection of a crude extract (MEF) prepared from cells of Listeria monocytogenes reduced the resistance of mice, guinea pigs, rabbits and rhesus monkeys to infection with Listeria. The resistance of mice to a number of other species of bacteria, both Gram-positive and Gram-negative, was also impaired by the cell material. The extract was heat stable when held at temperatures up to 70°C for 30 min. It was effective when administered prior or subsequent to infection or when injected by a route other than that used for challenge.

Injection of MEF into guinea pigs caused (a) a loss of weight, (b) changes in body temperature, (c) qualitative and quantitative alteration of the white blood cells, including monocytosis and (d) changes in serum lysozyme, properdin and complement.
The Inactivation of Rinderpest Virus by β-Propiolactone and Its Effect on Homologous Complement-Fixing and Neutralizing Antibodies
Authors: Stone S. S. , DeLay P. D.
Published: January 2023

Cattle inoculated with 5 × 104 lethal doses of Pendik rinderpest virus treated with 0.4% β-propiolactone (BPL) did not develop the disease and were susceptible to challenge. The same concentration of BPL also inactivated the Nakamura III strain of rinderpest. BPL in concentrations from 0.1 to 1.0% did not significantly alter the complement-fixing (CF) activity of anti-rinderpest rabbit serum. However, the titers as measured by agar gel diffusion (AD) were lowered. Rabbit anti-rinderpest serum treated with 0.4% BPL retained its virus neutralizing capacity when tested in rabbits. The CF and AD titers of Pendik virus were not altered after treatment with BPL in concentrations ranging from 0.1 to 1%.
Endotoxin Susceptibility and Delayed Hypersensitivity in Experimental Histoplasmosis
Authors: Box Edith D. , Briggs Norman T.
Published: January 2023

Mice with a sublethal Histoplasma infection were shown to have an increased susceptibility to a bacterial endotoxin (S. typhosa, lipopolysaccharide). This increase in susceptibility was directly related to the degree of stimulation of the RES by the infection as reflected by change in liver size. Delayed hypersensitivity and increased susceptibility to endotoxin are separate phenomena as judged by their temporal relationships to the course of a sublethal Histoplasma infection. Although susceptibility to endotoxin declined as the organism count declined, delayed hypersensitivity to the homologous challenge remained at a high level when most of the organisms had been eliminated. On the other hand, both phenomena were inhibited by chemotherapy of the infection, and specific hypersensitivity was decreased by treatment with endotoxin.
Immunochemical Properties of the Cross-Reactions between Anti-BSA and Heterologous Albumins
Authors: Weigle William O.
Published: January 2023

The immunochemical properties of the cross-reactions of 11 mammalian serum albumins with rabbit anti-BSA were investigated. The degree of the cross-reactions, as measured by precipitation, ranged between 5 and 75%. Nonmammalian albumin showed no cross-reaction with rabbit anti-BSA. In general, the initial combining ratios and the combining ratios at equivalence were lower with the heterologous reactions than with the homologous reaction. The precipitation curve obtained with the heterologous albumins showed the typical zones of excess antibody, equivalence and excess antigen. When I131 trace-labeled heterologous albumins were added to anti-BSA, virtually all of the I* activity was precipitated in the zone of excess antibody. The reaction between BSA and anti-BSA after absorption of the anti-BSA with sheep albumin gave a lower initial combining ratio and a lower ratio at equivalence than before absorption.

The addition of an extreme excess of heterologous albumin to anti-BSA previously absorbed with the same albumin, failed to inhibit markedly the precipitation of the remaining antibody by BSA. Only 2 to 14% more cross-reacting antibody could be detected by an inhibition test than could be detected by direct precipitation.

Studies employing absorption techniques, inhibition tests and double diffusion in agar, demonstrate that the cross-reactions between heterologous albumins and anti-BSA result from a sharing of certain related or identical determinant groups which represent only a portion of the total determinants present on the molecules. With absorption techniques and double diffusion in agar, it was shown that most heterologous albumins contain two or more serological types of determinants which crossreact with anti-BSA. Pepsin degraded BSA was shown to contain at least four serologically distinct determinants.
The Effect of Unheated Normal Bovine Serum on the Complement-Fixing Activity of Heat Inactivated Bovine Antiserum with Homologous Antigen
Authors: Rice Christine E. , Carrière J.
Published: January 2023

Fresh unheated normal bovine serum has been fractionated by dialysis against phosphate buffer, pH 5.0 to 6.2 and ionic strength 0.02, and the fractions tested for supplementing activity in complement fixation tests of two bovine antibody-antigen systems, an antibacterial and an antiviral. These bovine antisera in the dilution used did not fix guinea pig complement in the presence of homologous antigen but showed strong fixation when the guinea pig complement was made up with diluted unheated normal bovine serum.

The precipitates obtained from the normal bovine serum as dialysis proceeded for 1 to 24 hr showed an increase in supplementing titer whereas the supernatants became progressively less active. The nonspecific complement-fixing activity with tissue extracts or other antigens remained in the supernatant. Precipitates formed during 18 to 24 hr of dialysis were sometimes anticomplementary, particularly those from pools of sera from adult cattle.
Resistance of Hamsters in Infections with Histoplasma Capsulatum
Authors: O'Hern Elizabeth M.
Published: January 2023

Of 108 mature hamsters inoculated subcutaneously with 5 × 104 yeast cells of Histoplasma capsulatum all developed a disseminated infection. Dissemination appears to have occurred rapidly in all animals: spleens from each of the five animals sacrificed on the 2nd day following injection were positive by culture. Furthermore, during the 6-week period of study either spleen or liver, or both, yielded positive cultures in all instances.

Daily composite culture and tissue section indices rose gradually during the first 2 weeks of infection at which time they reached a plateau which was roughly maintained throughout the remainder of the 6-week test. Individual daily indices showed extensive variation throughout the plateau period.

Certain patterns of resistance and susceptibility were discerned among the responses of the individual animals to infection. These patterns ranged from apparent early resistance with minimal spread of the infection, to late resistance following extensive dissemination; and from extreme susceptibility resulting in death within the first 2 weeks of infection, to a progressive deterioration during the course of infection.

Evidence suggesting some degree of effective resistance was found in 51% of the animals infected 2 weeks or longer.

Large yeasts were found in 45% of the infected livers. In all such livers well circumscribed lesions were present.

Zones of inhibition of organism growth surrounding the tissue on culture medium were observed increasingly during the last 2 weeks of the investigation, as well as an increasing number of negative cultures of tissue found positive in tissue section. Possible bases for this growth inhibition and its relation to cultural diagnosis of human infection are discussed.
Studies on Immunity to Toxins of Clostridium Botulinum
Authors: Fiock Mary A. , Devine Leonard F. , Gearinger Naomi F. , Duff James T. , Wright George G. , Kadull Paul J.
Published: January 2023

The response of man to immunization with a purified, bivalent, AB botulinum toxoid was investigated. The toxoid was well tolerated; only occasional mild local reactions were encountered. Four schedules of immunization were studied; 0.5-ml injections were given at 0, 2, 4 and 6 weeks, at 0 and 8 weeks, at 0, 2 and 10 weeks and at 0 and 10 weeks; serum antitoxin titers were determined at intervals thereafter. Titers of 0.02 unit of type A and 0.005 unit of type B antitoxin were considered satisfactory, since guinea pigs with these antitoxin levels survived intraperitoneal i.p. challenge with approximately 105 mouse i.p. LD50 of homologous toxin. The 0-2-10 week immunization schedule produced the highest proportion of satisfactory titers, and the highest mean titers. Booster inoculations of 0.5 ml were given 1 year after the initial injection, except for one group in which boosters were given after 6 months. Marked increases in antitoxin titers were observed following the 1-year boosters, the mean titers reaching a value at least 500 times the satisfactory level and remaining well above this level for at least 2 years. Six-month boosters were considerably less effective, both with respect to the magnitude and to the duration of the secondary response. Prior immunization with univalent type A toxoid depressed the response to the type B component of the bivalent preparation. After storage for 3¼ years at 4°C the toxoid appeared to have deteriorated slightly, although considerable antigenicity remained.
Electron Microscopic Counting of Virus Particles by Sedimentation on Aluminized Grids
Authors: Smith Kendall O. , Melnick Joseph L.
Published: January 2023

The technique described employs collodion covered grids which have been coated with a thin layer of aluminum. Negatively charged virus particles are sedimented at high speed directly upon the positively charged aluminized grid surface. Grids may be washed under a stream of water to remove soluble salts and further purify poliovirus without detectable loss of virus particles. Particle distribution is satisfactory for counting with the electron microscope. Precision of counting is quite high when substantial numbers of particles are counted, and the number of particles counted per unit area is directly proportional to the relative concentration. The relative simplicity of the method permits numerous determinations to be made within a short period of time. The method has been used satisfactorily with poliovirus, bacteriophage and vaccinia virus, three viruses differing greatly in size and morphology.
Immunofluorescent Studies with Trachoma and Related Antigens
Authors: Nichols Roger L. , McComb Dorothy E.
Published: January 2023

Methods are presented for the utilization of immunofluorescence in field and laboratory studies of the PLT antigens: Antigen-antibody reactions produced easily discernible fluorescence of PLT viruses in yolk sac and tissue culture. Elementary body inclusions in conjunctival scrapings from the eyes of trachomatous patients yielded a brilliant stain. Evidence for the specificity of these reactions is discussed.The titration of PLT antisera by fluorescent antibody is presented.The superiority of fluorescent antibody to Macchiavello staining in the early detection of trachoma virus in egg isolation passages is detailed.A screening test for the presence of PLT antibodies in serum is described, based on the specific inhibition of immunofluorescence.The enhancement of serologic specificity of trachoma antiserum by cross-absorption with trachoma or LGV antigens is reported.

These studies, if confirmed by others, constitute another step in the serologic differentiation of the psittacosis-lymphogranuloma venereum-trachoma group of viruses.
Mumps Virus Replication
Authors: Treagan Lucy , Fong Jacob
Published: January 2023

Although 84 to 99% of seed virus was adsorbed to chorioallantoic membranes of chick embryos within 2 hr after infection, only 1% or less of the introduced virus was recoverable during the latent period. The length of this latent period was found to be at least 20 hr.

The development of hemagglutinins and viral complement-fixing antigens in general paralleled that of infective virus and a rapid release of virus and these viral components form cells was in evidence. The development of soluble antigen, however, showed a more specific relationship to infective virus and release of this component from cells (if indeed it did originate from cells) was considerably delayed.

The treatment of viral particles with anesthetic ether released internal S antigen; the quantity of released internal S antigen was related to the original concentration of the viral sample. The V antigen titers decreased 4- to 32-fold and hemagglutinin titers for chicken cells dropped 32-fold or more as the result of other treatment.
A Study of Macrophages and Epithelioid-Like Cells from Granulomatous (BCG-Induced) Lungs of Rabbits
Authors: Myrvik Quentin N. , Leake Eva Soto , Oshima Shunsaku
Published: January 2023

Techniques have been described for obtaining alveolar macrophages form granulomatous lungs in rabbits in large quantities. These macrophages demonstrate marked variations in their cytoplasmic volume suggesting that some of them may be epithelioid cells. Lysozyme analysis of the extracts indicate that the macrophages collecting in the lungs of rabbits undergoing an acute type of granulomatous response (vaccinated-challenged) contain less lysozyme than normal alveolar macrophages. In contrast, extracts from macrophages form the lungs of rabbits undergoing a chronic type of granulomatous response (vaccinated-not challenged) contain more lysozyme than normal alveolar macrophages. These data suggest that the cells in the acute type of response include cells of extrapulmonary origin which are low in lysozyme.
A Comparison between Cross-Tolerance and Cross-Reaction Induced by Bovine γ-Globulin
Authors: Timourian H. , Schechtman A. M.
Published: January 2023

The effect of neonatal injection of BGG on subsequent production of anti-BGG, anti-BSA and anti-HGG was studied in the rabbit. The magnitude of cross-tolerance was compared with the degree of cross-reaction between anti-BGG and BSA and HGG. Injection of 60 mgs of BGG into neonatal rabbits resulted in: a) 95% suppression of anti-BGG when the rabbits were challenged at 4 and 5 months of age; b) 70 to 90% suppression of anti-HGG production in comparison with noninjected littermate controls tested at 4 and 5 months of age; and c) 70% suppression of anti-BSA at 5 months of age. Anti-BSA production at 4 months of age was not significantly affected. Rabbit anti-BGG cross-reacted with BSA (6%) and with HGG (3%). The high degree of specificity of anti-BGG produced in normal adult rabbits is in contrast with the relatively low specificity of immunological tolerance induced by the same antigen in neonatal rabbits.
The Blood Interrelationship of Horses, Asses and Mules
Authors: Walsh L. S. N.
Published: January 2023

Since the discovery of normal human hemagglutinins by Ehrlich and the pointing out of their occurrence in groups by Landsteiner (1) with subsequent confirmation by Jansky (2), Moss (3) and many others, numerous workers have contributed to the more decisive identification of these groups. It has been shown by absorption as well as by specific agglutination (Hektoen (4), Koekert (5),) that four definite types of human blood can be identified.

Although iso-hemagglutinins are found in most species of lower animals it seems that no definite grouping exists. Ottenberg and Friedman (6) claim to have obtained definite typing in rabbits and steers.

Hektoen (7) studied the blood of not less than ten of each of the following species: rabbit, guinea pig, dog, horse and ox. He did not observe iso-hemagglutination in any of them.

Fishbein (8) studied 60 hogs, 60 cattle, 40 sheep, 25 rabbits, 10 dogs, and 20 frogs.
The Importance of Polyvalence in the Use of Therapeutic Antimeningococcus Serum
Authors: Felix A. , Yunowich R.
Published: January 2023

In view of the poor results obtained from specific serum therapy in sporadic cases of meningococcus meningitis in Jerusalem the meningococcus cultures which were isolated were subjected to a closer serological examination. We found that they did not seem to show similarity either with the English or with the American types. Moreover, the strains which we examined proved to be different with regard to their “receptor apparatus.” This may be attributed to the relatively great serological variability of the meningococcus.

Of all the therapeutic meningococcus sera which were examined, the ones from the Rockefeller Institute and from the Division of Laboratories and Research, New York State Department of Health proved to be by far the most effective in their action upon our cultures.

Therefore it seems that the serological examination of therapeutic meningococcus serum to be used in practice on the cultures isolated in various countries and in various epidemics is most urgent.

The authors wish to thank Dr. Wallach of the Shaare Zedek Hospital in Jerusalem for his coöperation in the carrying out of this work.
Acute Inflammation Caused by Antibody in an Animal Previously Treated with Antigen
Authors: Opie Eugene L.
Published: January 2023

Normal rabbits may be sensitized to the local action of horse serum by injection of serum obtained from an animal immunized by repeated injections of the antigen. Horse serum injected into the dermis of this passively sensitized animal causes acute inflammation (Arthus phenomenon). This passive transfer of sensitization shows that the blood serum of the immunized animal contains an antibody which introduced into a normal animal renders it sensitive. Furthermore, simultaneous injection of horse serum and anti-horse serum from rabbit causes acute inflammation which fails to occur when anti-horse serum is replaced by normal serum.

These observations suggest the possibility that acute inflammation will occur whenever antigen and antibody come into contact within the tissues. The attempt has been made to determine whether a normal rabbit which has received horse serum will react with acute inflammation when serum from an immunized or “sensitized” animal is injected into its tissues.
The in Vitro Detection of “Cell-Fixed” Hemagglutinating Antibodies to Tuberculin Purified Protein Derivative (PPD) in Humans
Authors: Freedman Samuel O. , Turcotte R. , Fish A. J. , Sehon A. H.
Published: January 2023

Extracts of the washed peripheral blood leukocytes of all adult human subjects studied were found to contain hemagglutinating antibodies to tuberculin purified protein derivative (PPD). Subjects with positive tuberculin skin tests had significantly higher titers of these “cell-fixed” hemagglutinating antibodies to tuberculin PPD than did tuberculin skin test negative subjects. The finding of low titers of “cell-fixed” antibodies in tuberculin skin test negative subjects may reflect minor degrees of tuberculin hypersensitivity not detected by skin testing, or may be due to cross-reactivity with Mycobacteria other than Mycobacterium tuberculosis. The “cell-fixed” antibodies are probably adsorbed from the serum of the leukocyte donor. No hemagglutinating antibodies to tuberculin PPD were found in sera or leukocytes obtained from the umbilical cord blood of newborn infants. The relationship between “cell-fixed” hemagglutinating antibodies to tuberculin PPD and “transfer factor” was not established in this investigation.
Immunochemical Study of Rabbit γ-Globulin Allotypes
Authors: Leskowitz Sidney
Published: January 2023

Antibodies to γ-globulin allotypes were produced in rabbits by immunization with purified rabbit γ-globulin. These antibodies were studied quantitatively by precipitin reaction and shown to be directed towards antigenic determinants occurring on the antibody combining Fragments I and II obtained by papain digestion of rabbit γ-globulin.

Similar allotype specific antibodies could be produced in goats by immunization with one type of rabbit γ-globulin and absorption with another type. One such goat antiserum was found to be capable of precipitating with whole rabbit γ-globulin of the appropriate allotype but not with the papain digest fragments obtained therefrom. Such fragments were, however, shown to inhibit precipitation with whole rabbit γ-globulin, thus indicating their apparent univalence.

When rabbit γ-globulin in the form of a specific precipitate in Freund adjuvant was used for immunization, an antibody specific for human γ-globulin was found in addition to the rabbit allotype specific antibody.
Immunochemistry of Coagulated Antigen-Antibody Complex
Authors: McDuffie Norman M. , Sutherland G. Bonar
Published: January 2023

Under suitable conditions, a number of chemical agents and/or heat may be used to render insoluble either antibody molecules or specific complexes, without destroying or masking all of the native combining sites. These coagula may be used as highly specific adsorbents for native antigen and the antigen may be subsequently eluted at pH 3.0. A study of some suitable coagulating reagents and conditions has been presented.
Antigenicity of Polypeptides (Poly-α-Amino Acids)
Authors: Maurer Paul H.
Published: January 2023

It has been shown that copolymers of glutamic acid and lysine having ratios of 6:4, 7:3 or 5:5 are antigenic in about 40 to 50% of Hartley strain guinea pigs. A 6:4 copolymer of glu-lys having a molecular weight as low as 5000 was also antigenic. Within the dosage limits studied, the amount of antigen injected in complete Freund's adjuvant did not appear to affect significantly the immune response. Both “immediate” and “delayed” type skin reactions were noted in the guinea pigs. With the hyperimmune sera injected intraperitoneally, recipient animals did not exhibit either the “immediate” Arthuslike or “delayed” reactions. However, these same sera could evoke good passive cutaneous anaphylaxis (PCA) reactions and sensitize guinea pigs for passive systemic anaphylaxis. Some speculations have been advanced to explain the behavior of the glu-ala and glu-tyr copolymers as well as the poor over-all reactions with the glu-lys copolymers.

The order of decreasing sensitivity in the test for the production of antibody was systemic anaphylaxis, PCA and “active intracutaneous anaphylaxis of the ‘immediate’ type.” With the systems employed the molecular weight of the glu-lys copolymer had little effect on the cross-PCA reactions when compared on a weight for weight basis.
Allergic and Immunologic Studies of a “Purified” Fraction of Ragweed Pollen
Authors: Arbesman Carl E. , Reisman Robert E. , Bonstein Herbert S. , Rose Noel R.
Published: January 2023

Summary and Conclusions

The allergenic and antigenic properties of pool C, a “purified” fraction of ragweed pollen, were investigated. As compared to whole ragweed pollen extract, pool C was found to be highly antigenic as measured by its ability to stimulate antibody production in rabbits, induce passive cutaneous anaphylaxis and provide indirect hemagglutination with both human and animal antisera. Pool C is not immunochemically pure as evidenced by three or four lines produced by its homologous antisera in immunoelectrophoresis. However, many of the antigens in the whole ragweed extract have been eliminated. Positive hemagglutination titers with pool C were found with the sera of 72% of untreated ragweed-sensitive individuals. This contrasted greatly with 20% positive reactions found with whole ragweed extract. On a weight basis, pool C was 100 to 1000 times more skin-reactive than whole ragweed extract, both by direct and passive transfer tests. In vitro and particularly in vivo neutralization tests indicated that some of the allergens in whole ragweed extract have been eliminated in the preparation of pool C.
Dissociation of Phagocytosis and Intracellular Killing of Staphylococcus Aureus by Human Blood Leukocytes
Authors: Li Isabel W. , Mudd Stuart , Kapral Frank A.
Published: January 2023

The sera of healthy adult humans contain thermostable and thermolabile components which promote the phagocytosis of Staphylococcus aureus by human leukocytes. Serum heated to 56°C for 7½ to 30 min produces phagocytosis at a somewhat slower initial rate; the ingested staphylococci are not killed intracellularly, at least within the 150 min of the present experiments. Unheated sera produce phagocytosis at a more rapid initial rate, and ingested staphylococci are killed intracellularly although killing is never complete within the time of these experiments. The amount of fresh serum required to produce this killing effect is very small, of the order of 0.05%.

Fresh serum also causes adherence of staphylococci to erythrocytes. Confusion due to this adherence phenomenon can, however, be obviated by appropriate arrangements of the experiments.
Studies on the Sensitizing Properties of Human Antisera and Purified Antibodies
Authors: Kabat Elvin A. , Liacopoulos Panayotis , Liacopoulos-Briot Monique , Halpern Bernard N. , Relyveld Edgar H.
Published: January 2023

Purified human antidextran, an S = 7 γ-globulin, was about 1000 to 3000 times less potent in sensitizing intestine in vitro and 25 to 100 times less effective in sensitizing for passive cutaneous anaphylaxis than were the most potent human diphtheria antitoxic sera under the conditions studied. Human antitoxic sera also varied as much as 100-fold in their sensitizing capacity for intestine/µg antitoxin N.
Normal Human Serum Derivatives That Inhibit the Tanned Cell Hemagglutination Test for Rheumatoid Arthritis and Other Reactions
Authors: Bloomfield Norman
Published: January 2023

With the exception of albumin, the major Cohn plasma fractions and the derivatives of normal human serum obtained by ultracentrifugation in high density salt solutions and by electrophoresis on Geon inhibit the tanned cell reaction with rheumatoid serum. Native γ-globulin is a poor inhibitor but heating increased its inhibitory activity 4000-fold. The most potent inhibitors were those rich in low-density (β) lipoprotein; lipoprotein-poor α- and β-globulins also possessed appreciable inhibitory activity. A large proportion of inhibition found in normal serum was recovered in the middle fraction obtained by ultracentrifuging β-lipoprotein-depleted serum in a 1.21 density salt solution; a slowly sedimenting (small molecule?) component of great inhibitory activity may therefore also be present.

Inhibition by the unheated fractions is not specific for the rheumatoid reaction; serum and several of its ultracentrifugal and electrophoretic fractions were examined and found to inhibit the tanned cell test for trichinosis. Inhibition by serum and several derivatives of lectin agglutination of sheep cells further demonstrated the nonspecificity of the inhibition.
Studies on the Immune Globulins of Human Serum
Authors: Heremans J. F. , Vaerman J.-P. , Vaerman C.
Published: January 2023

The serum of a subject vaccinated with diphtheria toxoid and two sera from patients recovering from infections with Brucella abortus suis were fractionated and samples consisting of γss- (7 S γ), γ1M- (19 S γ) and γ1A- (β2A-) globulins were separately assessed for antibody activity. All three protein species appeared to be serologically active.

Particular care was taken with the interpretation of data concerning the γ1A-globulin samples. The serologic activities of the γ1A samples could not be explained on the basis of any contamination with γss-globulin, and were ascribed to γ1A-globulin itself. A peculiar feature of the γ1A anti-Brucella agglutinins here studied was their absence of complement-fixing capacity.

It is suggested that future investigations on the physicochemical nature of antibodies be carried out with the knowledge that γ1A- (β2A-) globulin constitutes a third class of antibodies, besides the γss- (7 S γ) and γ1M- (19 S γ) classes.