bluuebunny/crossref_metadata_2025 huggingface dataset:
Neonatal Rabbits as Donors of Transferred Lymph Node Cells
Authors: Harris Susanna , Harris T. N. , Ogburn Clifton A. , Farber Miriam B.
Published: January 2023

Young rabbits were used as donors of lymph node cells for incubation in vitro with Shigella-trypsin filtrate and transfer to irradiated recipient rabbits. No agglutinins to Shigella could be detected subsequently in the sera of the recipient animals if the donors were less than 1 month of age. Thereafter, a progressive rise in maximal antibody level of recipients was found with increasing age of the donors, up to an age of approximately 3 months. Donor rabbits injected with Shigella in the 1st month of life yielded cells which, again, were ineffective. Thereafter, the effectiveness of the cells increased substantially more rapidly with increasing age of donor than in the case of antigen-incubated cells.

On active “immunization” of young rabbits with Shigella-trypsin filtrate at an adequate dosage level, agglutinins were detected only after the 1st month of life. With larger doses of this antigen, the antibody appeared in younger rabbits, but at a later time relative to the injection of antigen and at a relatively low titer. For comparison, other young rabbits were injected with alum-precipitated bovine γ-globulin. Anti-BGG could be detected in the sera of some rabbits injected as early as 3 days of age, and regularly in rabbits injected after the 1st week of life.
Studies on the Coliphage Neutralizing Activity of Normal Human Serum
Authors: Cowan Keith M.
Published: January 2023

The ability of normal human serum to neutralize the coliphages T2, T4, T6 or T7 was either abolished or markedly depressed by heat treatment, decomplementation, absorption with zymosan, or removal of divalent cations from the serum. The phage neutralizing activity of normal serum was inhibited by irradiated phage preparations. The inhibition produced was always greater in homologous systems than with heterologous ones. This would indicate that the normally occurring phage neutralizing substances exhibit some degree of serologic specificity.
Antibody Response of Adult Chickens to Infectious and Noninfectious Foot-and-Mouth Disease Virus
Authors: Fellowes O. N.
Published: January 2023

Lesions suggestive of infection were observed in or on the mouth, combs and feet of chickens inoculated with Type A-119 virus propagated in tissue culture. Lesions were not observed in chickens inoculated with Type A-GB virus propagated in guinea pig footpads.

A high titer of neutralizing antibody was induced in adult chickens by inoculation of 1 and 2 ml of either infectious or noninfectious Type A-119 virus grown in tissue culture. Infectious Type A-GB, propagated in guinea pig pads, produced a relatively low titer of neutralizing antibody in chickens.

Both strains, 119 and GB, of infectious Type A virus produced a good response of complement-fixing antibody in chickens as tested by a modified direct complement fixation test.

No evidence of infection was found in normal chickens held with infected birds in the same cage.
Quantitative Analyses by Diffusion Column Reactions
Authors: Glenn William G.
Published: January 2023

Experimental interpolation of slope K (diffusion (mm) vs √min) for SD columns of albumin homologous reactions resulted in the detection of concentration differences of 0.10 mg N2/ml (0.06% protein) of antigen and 0.010 mg N2/ml of antibody in 75% and 50% of the respective paired slope comparisons within specific antigen and antibody ranges. Comparison of the Kjeldahl concentrations with the same concentrations estimated from diffusion measurements for both antigen and antibody strengths in SD columns were made using results from regression of K on log of the appropriate reactant concentration. The data used were strictly selected for linearity and reproducibility. For 30 antigen derivations involving Fractions II and V homologous systems at three antibody levels, the mean deviation was 3.44%. The smallest deviation was -0.29% or 0.001 mg antigen N2 lower for the predicted as compared to the Kjeldahl concentrations. Estimated homologous antibody concentrations from rigidly selected data and curves (K regressed on the logarithm of the antibody concentrations), when compared with Kjeldahl values, showed an average deviation of 2.54% for 50 antibody derivations at five antigen levels for Fractions II and V in SD columns. The over-all mean difference (sign ignored) between Kjeldahl and diffusion—estimated values—was 0.007 mg antibody N2/ml. The limits of resolution for reactant concentration differences in SD column reactions at certain ranges are discussed in detail.
Virus-Tissue Union and the Pathogenesis of Enterovirus Infections
Authors: Kunin Calvin M.
Published: January 2023

A heat labile, sedimentable substance capable of inactivating Coxsackie B1 and B3 viruses has been demonstrated in noncultured mouse brain and in certain other murine and primate tissues. The tissue and age distribution of this factor appears, in general, to be correlated with the in vivo sites of multiplication of these viruses and with their virulence for newborn and adult mice. It has been postulated that this material in brain and other tissues is specific virus receptor-site and that the relative abundance of this substance in cells of various tissues is an important determinant of age and tissue specific susceptibility to these agents.

Sequential loss, with age, of adsorptive capacity of mouse brain for Coxsackie B1 virus was demonstrated in quantitative studies. Amount of tissue employed was rate limiting, but inoculum size was not critical. Cortisone treatment did not increase adsorptive activity of adult mouse brain for this virus. Adsorption of Coxsackie B1 and poliovirus by brain homogenates was temperature dependent, being more marked in the case of the former. Adsorption of poliovirus by noncultured, extraneural rhesus monkey tissues was confirmed; adsorption of Coxsackie B1 virus was limited to rhesus monkey cerebral cortex, cerebellum and liver. Brain weight-virus adsorption curves demonstrated about equal activity of rhesus monkey brain against both virulent and avirulent polioviruses but marked differences were observed in similar studies utilizing newborn mouse brain against virulent and avirulent strains of Coxsackie B3 virus.

These studies are presented as evidence that age specific susceptibility and mouse virulence of certain Coxsackie viruses are related to the relative abundance of receptor-like substance in host tissues.
The Growth Cycle of a Trachoma Agent in FL Cell Cultures
Authors: Bernkopf H. , Mashiah P.
Published: January 2023

FL cell cultures were infected with a Chinese strain of trachoma, and the development of infectivity during a single growth cycle was studied. A noninfective phase lasting at least 24 hr after infection was demonstrated. Infectivity reappeared at 28 hr and reached a maximum at 48 to 72 hr. These findings agree well with the morphological findings reported before, according to which particles of elementary body size changed their appearance shortly after infection, could again be demonstrated at 28 hr, and then rapidly increased in number.
Growth Characteristics of Respiratory Syncytial Virus
Authors: Jordan William S.
Published: January 2023

Respiratory syncytial virus was studied with respect to the effect of type of cell and composition of nutrient fluid on cytopathic manifestations and multiplication in tissue culture. Production of syncytium was striking in HEp-2 cells, minimal in KB and HeLa cells; rounding was the characteristic response of DMB cells. In comparison with a maintenance fluid containing Eagle's minimal essential medium, a fluid utilizing Scherer's solution limited the formation of syncytium and delayed the evolution of cytopathic effect, particularly in HEp-2 cells. The presence or absence of syncytium had little effect on the total yield of infectious virus.

The latent period of the initial multiplication cycle was 12 to 14 hr, and was followed by rapid growth and release of virus. A method for cell disruption which released cell-associated virus with little inactivation was utilized to demonstrate that nearly 50% of virus is still cell-associated even when the cytopathic effect is marked. Some virus was inactivated by a single freeze-thaw cycle, but aliquots stored at -70°C maintained stable and adequate levels of infectious virus for at least 6 months.
Accelerated Breakdown of Immunological Tolerance Following Whole Body Irradiation
Authors: Mäkelä O. , Nossal G. J. V.
Published: January 2023

Rats were made unresponsive to mouse erythrocytes or bovine serum albumin by a series of injections starting at birth. The unresponsiveness, though not always complete, remained constant in a given animal if antigen injections were continued. In animals irradiated with 550 or 600 r the tolerance usually broke down, with the formation of substantial amounts of antibody.

Normal animals also irradiated at a similar age and given repeated injections of mouse erythrocytes formed excess antibody during the 5th to the 13th postirradiation week. By contrast, irradiated rats receiving a single injection of bovine serum albumin in Freund's adjuvant formed less antibody than did nonirradiated controls. The theoretical implications of the work are discussed.
Precipitating and Nonprecipitating Antibodies in the Primary and Secondary Immune Responses
Authors: Blumer H. , Richter M. , Cua-Lim F. , Rose B.
Published: January 2023

The formation of antibodies to soluble protein antigens for many months after primary immunization has been confirmed. These antibodies were produced after the immunization of rabbits with bovine serum albumin, egg albumin, human γ-globulin, pig γ-globulin and bovine γ-globulin. The antibodies present in the circulation 11 to 16 days after the initial immunization could be detected by using antigen-coated red cells and, in most cases, by the precipitin technique as well. Those antibodies present in the circulation several months later could be detected only by the hemagglutination technique. The hemagglutinating antibody titer remained constant for at least 6 months after primary immunization. In some cases the nonprecipitating antibody concentration was found to rise at a time when the precipitating antibody level was declining. Hemagglutinating antibody titers of sera, following secondary immunization, rose to high levels very rapidly and then started to decline, rapidly at first and more slowly later. Only those antisera which were obtained during the early period of antibody production and which contained precipitating antibody could passively sensitize guinea pigs. Cortisone acetate inhibited the primary response to intravenously injected antigens in six out of seven rabbits. It failed, however, to inhibit the secondary antibody response.
Immunochemical Studies on Bacteriophage Deoxyribonucleic Acid
Authors: Murakami William T. , Van Vunakis Helen , Lehrer Harris I. , Levine Lawrence
Published: January 2023

Several rabbit antisera to ruptured T2, T4 and T6 bacteriophage contained antibodies to DNA. With all of the antisera, denatured DNA was more effective serologically than native DNA. Of 41 different DNA preparations tested with an antiserum to T4 DNA, only DNA isolated from T2, T4 or T6 bacteriophage fixed C′. The specificity of the antibodies as measured by cross-reactivity is described. The antigenic determinant groups of T-even bacteriophage DNA include, in part, the glucosyl residue of the DNA.
The Staining Properties of Human Serum Proteins Conjugated with Purified Fluorescein Isothiocyanate
Authors: Frommhagen Laurence H. , Spendlove Rex S.
Published: January 2023

Electrophoretic analysis has shown that six preparations of fluorescein isothiocyanate (FITC) from one source contain degradation products which exhibit essentially similar fluorescence properties and a similar capacity to couple with serum protein, but which confer upon the conjugate the propensity for nonspecific staining. It is recommended that only undegraded FITC (e.g., BBL) which manifests superior specificity and higher fluorescence intensity, be used for the conjugation of specific immune γ-globulins, and that the pH of the conjugation system should not exceed 8.8. Purified FITC is now commerically available or may be easily obtained by continuous flow electrophoresis. It is emphasized, however, that the degradation products of FITC constitute only one of a number of factors contributing to nonspecific fluorescent staining. Two other such factors, relating to the overlabeling of the γ-globulin and to the other serum proteins, are discussed.
Evaluation of the Complement Method of Fluorescent Antibody Technique with Myxoviruses
Authors: Hinuma Yorio , Ohta Rei , Miyamoto Tsutomu , Ishida Nakao
Published: January 2023

Serum globulin obtained from rabbits which had been immunized with the globulin fractions of guinea pig sera revealed anti-C′ activity. When this rabbit globulin was conjugated with fluorescein isothiocyanate, the sedimentation pattern and constant were unchanged, and no decrease of anti-C′ activity followed. The molecular ratio of the dye to globulin was found to be approximately 4:1.

The higher sensitivity of the C′ method, in comparison with the indirect method, was established quantitatively in detecting both antigen and antibody of Sendai virus. Moreover, more intense staining was obtained by the C′ method.

Application of the C′ method to detect the antigens of influenza A and B viruses in tissue cultured cells, as well as those of NWS virus in Ehrlich ascites tumor cells, showed good correlation with the results obtained with Sendai virus. Antibody titers obtained by this method were similar in magnitude to those obtained in in vitro CF tests.
Sensitization and Desensitization of Mice to Histamine and Serotonin by Neurohumors
Authors: Fishel Charles W. , Szentivanyi Andor , Talmage David W.
Published: January 2023

The urinary elimination pattern of normal mice following the injection of radioactive histamine and serotonin was compared with that of pertussis-treated animals. The absence of gross differences between these two groups suggested that the handling of histamine and serotonin by the body as a whole is not altered in the highly sensitive pertussis-vaccinated mouse. For this reason, it was concluded that the hypersensitivity is the result of a localized hyper-reactivity of the effector cells to normally nontoxic concentrations of histamine and serotonin. Attempts were therefore made to modify the sensitivity with a variety of neurohumoral agents known to react with the receptors of the neuroeffector cells.

It was found that acute exposure to progressively increasing amounts of histamine and serotonin desensitized the pertussis-treated animals to these amines and the desensitization persisted for at least 24 hr. Cross-protection was also obtained; complete with histamine pretreatment against serotonin, incomplete with serotonin against histamine. However, desensitization attempts with another toxic amine (agmatine), not known to function as a neurohumor and whose toxicity for mice is not modified by pertussis-sensitization, demonstrated that the resistance of either normal or pertussis-treated mice to the same amine was not altered, nor was this procedure of any apparent value in protecting the latter animals against histamine.

The agmatine disparity further emphasized the critical significance of neurohumors in the pertussis-induced sensitive state and led to the administration of various autonomic drugs which were shown to be protective in the pertussis-treated animals. Another such substance, dibenzyline (α-adrenergic blocking agent), was also protective in pertussis-sensitized animals, whereas a β-adrenergic blocking agent, dichloroisoproterenol, duplicated the histamine and serotonin hypersensitivity in normal animals.

Based on these observations, the hypothesis is advanced that the pertussis-induced hypersensitivity is the result of a functional imbalance between two types of adrenergic effector systems, i.e., between two types of adrenergic receptors or in the neural pathways leading to them.
Immunization in Wasp-Sting Allergy through Venom-Repositories and Periodic Insect Stings
Authors: Loveless Mary Hewitt
Published: January 2023

Summary and Conclusions

Of 11 persons with alarming immediate reactions to polistes stings, 9 have been immunized with Arlacel:petrolatum:venom emulsions during 1960 and 1961, a total of 12 such annual treatments being given the group. For all but one member (BA), the repository was comprised of 1.2 ml containing all the venom of six live polistes workers, placed under the skin in a single injection (patient AT preferred a divided dose). There were no adverse effects either focally at the time of therapy or locally during the ensuing year or two. Seven of these members had been immunized, in one or more preceding years, with the same annual dose administered in fluid form during a one-visit, 2½-hr course that employed five to seven endermal injections. Five of their 19 treatments with fluid venom had, in contrast to the fully tolerated repositories, evoked borderline focal signs. In addition to these dually-treated patients, there was one physician who had tolerated five annual, 6-sac endermal courses of fluid venom, and another man who accepted uneventfully the full standard dose of venom in emulsified form.

Repositories built up a satisfactory degree of immunity as judged by six planned and two accidental stings by one to three polistes inflicted on eight members at intervals as great as 7½ months after treatment. Only two trivial focal reactions were engendered. Resistance appeared to be equally good after the standard series of fluid venom injections, for there was only one trivial adverse development among 16 challenges that were made as soon as 13 days or as late as 41½ weeks after therapy.

Emulsification decreased the venom's capacity to evoke local and focal responses. There were no cysts, abscesses, or new-growths at the repository sites.

In an effort to provide year-round protection, 8 of the 11 members have, since their exhibition of tolerance toward stinging wasps, been transferred 2 to 21 weeks later to a regimen of intermittent stinging. Five trivial and one mild focal response have been engendered by 41 such sting treatments. The latter serve not only as probable booster stimuli but also as periodic tests of immunity.
Immunization against Parainfluenza Infections
Authors: Jensen Keith E. , Peeler Betty E. , Dulworth William G.
Published: January 2023

Following adaptation of strains of parainfluenza types 1 and 3 to embryonated eggs, antigenic comparisons were made with convalescent ferret sera and postvaccination human sera. Homotypic reactions evident with ferret sera were not confirmed by responses of man which indicated a close antigenic relationship between the Sendai and HA-2 strains of type 1 and also heterotypic crossing with type 3 strains, SF-3 and HA-1.

Vaccines prepared by formalin treatment of virus sedimented from infected egg fluids have been found to be safe and potent in adults, children and infants. Possible explanations have been discussed for the relative poor antigenicity of type 1 virus.
Increased Precipitability of Antibody as a Result of Iodination
Authors: Pressman David , Radzimski Gerald
Published: January 2023

The precipitability of anti-p-azobenzoate antibody by an antigen containing azobenzoate groups is not complete and soluble complexes of antigen-antibody are present in the supernatant. A limited degree of iodination of the antibody increases its precipitability by antigen. Extensive iodination causes decreased precipitation presumably through effect on the binding site itself. The increase in precipitability is explained on the basis of increased hydrophobic bonding, or increased van der Waals' interaction due to the added iodine atom or denaturation.

The use of chemically altered antibody systems is proposed for a study of the effect of alteration on protein-protein interactions in aqueous systems. The precipitability of antiprotein antibody such as anti-BSA or antibovine γ-globulin antibodies is markedly decreased at levels of iodination which give increased precipitation with antihapten antibody. It would appear that the antiprotein antibody site is more sensitive to iodination than antihapten antibody sites. This may be due to presence of a particularly important and sensitive group in the site or due to the presence of more than one iodinatable group in the site, any one of which when iodinated destroys ability of the site to combine with antigen.
Inhibition of Complement Fixation by Human Serum
Authors: Heimer Ralph , Levin Francoise M. , Primack Anita , Corcos Josue M. , Nosenzo Carlo
Published: January 2023

Assays for inhibition of fixation of gpC′ were described. The inhibitory activity of human sera was found to vary, but inhibitory activity was significantly elevated in sera of individuals having rheumatoid arthritis.

Fractionation of a number of sera of healthy donors by DEAE cellulose chromatography revealed two inhibitors of CF. In addition to these substances, a third compound occurred in the sera of individuals with rheumatoid arthritis. The compound was purified 400-fold and was shown to be a γ-macroglobulin and was distinct from RF.
Leukocyte Antibodies in Inbred Strains of Guinea Pigs Following First- and Second-Set Skin Homografts
Authors: Walford Roy L. , Anderson Robert E. , Carter P. Kenneth , Mihajlovic Felicia
Published: January 2023

The development of leukocyte antibodies in inbred strains of guinea pigs following skin homografting in demonstrated. These antibodies react strongly with lymphocytes and only weakly, or not at all, with neutrophils. Following repeat set-grafts in guinea pigs, a prompt and marked rise in leukocyte antibody titer takes place, suggesting an anamnestic response. The production of humoral antibodies reactive with leukocytes (lymphocytes) may be a general phenomenon following homografting in all species. The possible significance of such a phenomenon in relation to transplantation immunity and transplantation disease is discussed.
Immunochemical Study of Antigenic Specificity in Delayed Hypersensitivity
Authors: Leskowitz Sidney
Published: January 2023

Heating, iodination, dinitrophenylation and disulfide bond cleavage, in increasing order, were found to reduce the ability of bovine serum albumin and ribonuclease to precipitate with rabbit antibody. A parallel decrease was noted in the ability of these modified antigens to produce anaphylactic sensitivity in guinea pigs to the native antigens.

Ability to produce delayed hypersensitivity in guinea pigs to native antigens was markedly diminished in all cases.

It was concluded that no dissociation of antigenic properties in respect to precipitation with antibody and production of anaphylactic or delayed sensitivity was produced by these alterations.
Quantitative Study of the Combination of Lima Bean Lectin with Human Erythrocytes
Authors: Boyd William C. , Bhatia Harkishin M. , Diamond Michael A. , Matsubara Shoji
Published: January 2023

The combination of lima bean anti-A lectin with human erythrocytes was studied quantitatively by use of lectin labeled by coupling with diazotized p-iodoaniline containing I131. The anti-A consistently took up two to four times as much I131 as did the nonspecific proteins present. It was found that the labeled lima bean proteins combined with erythrocytes in greater and greater amounts as the ratio of lectin to cells was increased. The relation between amount of lectin per cell and ratio of lectin to cells added was found to be substantially linear if both were plotted logarithmically, so that the amount of lectin per cell did not reach a maximum in the range studied. In addition to the anti-A which combined with group A red cells and agglutinated them, there was present in the lectin preparations large amounts of a protein that combined with all human erythrocytes without agglutinating them. Homozygous A1A1 cells seemed to combine with more anti-A than did heterozygous A1O cells. A2 cells may combine with somewhat less, but on the whole the results suggest that the main difference between A1 and A2 is qualitative rather than quantitative. A3 cells combined with much less anti-A lectin than did A1 or A2 cells.

That the extra radioactivity taken up by group A cells was due to the combination of anti-A lectin with them was shown by the fact that N-acetyl-D-galactosamine would inhibit this combination while leaving the amount of nonspecific protein combined unchanged, and also by the fact that after the labeled lectin had reacted with A substance it no longer added more radioactivity to A cells than to O or B cells.

If it is assumed that each molecule of the anti-A lectin that combines with a group A erythrocyte is combined with an A combining site, we can calculate, making plausible assumptions as to the molecular weight of the lectin, how many such A-specific sites there are in a group A cell. The results are astoundingly high, as much as 8 or 10 million. Nevertheless, it is thought that these figures may not be in error, though possibly they may refer to the A-combining sites in the entire stroma rather than merely to those available on the surface.
In Vitro Uptake of Antigen-Antibody Complexes by Phagocytic Cells
Authors: Patterson Roy , Suszko Irena M. , Pruzansky Jacob J.
Published: January 2023

The ingestion of antigen-antibody complexes prepared as soluble materials was studied by a method of in vitro uptake by phagocytic cells using I131 trace labeled complexes. Different degrees of in vitro pinocytosis occurred with preparations of complexes at different degrees of antigen excess. This in vitro pinocytosis was also demonstrated by a fluorescent antibody technique localizing the complexes intracellularly. These methods provide a system of study of transfer of soluble materials across the cell membranes of phagocytic cells which can be compared with the same phenomenon in vivo. The in vitro uptake of complexes was compared with antigen binding to globulin by the ammonium sulfate technique and the in vivo localization of radioactive trace labeled antigen-antibody complexes. By these methods the ingestion of antigen-antibody complexes by cells may be studied in a simplified manner. The localization of antigen-antibody complexes in phagocytes occurs both with complexes prepared with rabbit or human antiserum against serum protein antigens.
On the Nature of the Precipitate in the Generalized Shwartzman Reaction
Authors: Berken A. , Wolman M.
Published: January 2023

Evidence has been presented which indicates that the generalized Shwartzman reaction in rabbits is primarily a consequence of the intravascular precipitation of serum proteins. These precipitable proteins are present in sufficient quantity in the normal animal to result in the generalized Shwartzman reaction soon after the other necessary conditions are invoked. Apparently, these conditions are the presence in the circulation of a sufficient amount of a precipitating substance and depression of reticuloendothelial system (RES) phagocytic function. The administration of relatively large amounts of a precipitating substance (Liquoid) seems to obviate the requirement for an exogenous RES depressant. Pretreatment with the RES depressant lipid, ethyl stearate, followed by the administration of a single dose of endotoxin results in the occurrence of the generalized Shwartzman reaction. The analogous effects of ethyl stearate and cortisone on the RES and their possible significance are discussed.
Delayed Cutaneous Reactions in the Rat
Authors: Flax Martin H. , Waksman Byron H.
Published: January 2023

Typical delayed skin reactions to tuberculin were elicited in rats sensitized by footpad injection of killed tubercle bacilli in oil. Their similarity to classical delayed reactions in guinea pig or rabbit is demonstrated by their identical time course, characteristic histologic appearance and passive transfer with sensitized lymph node cells. Similar reactions were obtained after sensitization with Nocardia asteroides in oil, OT in incomplete Freund's adjuvant or BSA in complete adjuvant. These were indurated reactions made up largely of mononuclear infiltrates in the deep dermis and subcutaneous tissues, maximal at 24 to 48 hr. This location of the lesions is correlated with the distribution of vessels of venous type.
Demonstration of a Toxin in Mumps Virus and Its Dissociation from Infective and Hemagglutinating Properties
Authors: Wacker Waldon B. , Lipton Murray M. , Steigman Alex J.
Published: January 2023

In controlled experiments, a hitherto underscribed lethal viral toxin has been demonstrated, employing neonatal mice intravenously injected with mumps vaccine. Unlike other viral toxins, in the system employed, toxicity is demonstrable with formalin inactivated mumps vaccine, as well as with infective preparations. Although related to initial viral concentration and age of the vaccines used, the toxicity observed is a property distinct from the hemagglutinin or infectious content of the preparations used. The observed toxicity is prevented by mumps antisera, but not appreciably by normal sera. Intravenous injection of neonatal mice may be a valuable tool for the study of viral toxins.
Precipitation of Radiolabeled Poliovirus with Specific Antibody and Antiglobulin
Authors: Gerloff Robert K. , Hoyer Bill H. , McLaren Leroy C.
Published: January 2023

A highly sensitive and specific serologic test for type 2 poliovirus or its antibody is described. Radiolabeled virus in submicrogram quantities was incubated with specific antiserum; thereafter, rabbit antiglobulin was added and the incubation repeated. Coupling between antiglobulin and globulin resulted in precipitation of both, along with virus in combination with immune globulin. Precipitation of radioactivity was used as the measure of virus-antibody reaction, since it paralleled precipitation of infectious particles. Thus the test has been called the radioisotope precipitation (RIP) test.

Sensitivity of the test varied inversely with concentration of virus and could be increased to about 80 times that of the tissue culture neutralization test. Ratio of antiglobulin to total globulin was unimportant if antiglobulin were in excess; likewise, quantity of precipitate formed was inconsequential if all of the specific immune globulin were included in it. The test could be completed in 2 to 3 hr, if necessary, and was performed satisfactorily with virus inactivated by carefully controlled ultraviolet irradiation. Inhibition of reaction between labeled virus and its antibody by prior exposure of the antibody to unlabeled virus resulted in conditions whereby unlabeled virus in 0.001 to 0.1 µg quantities could be assayed. Although most of the studies were done with P32-labeled virus, the feasibility and advantages of using I131 for labeling were investigated. Removal of infectious units from the supernatant was also a satisfactory index of presence of specific antibody and enables the use of minute quantities of nonradioactive, biologically active antigen in this type of test.
Studies on Japanese B Encephalitis Virus Vaccines from Tissue Culture
Authors: Rohitayodhin Skon , Hammon W. McD.
Published: January 2023

Experiments were performed on the cultivation of the Nakayama strain of JBE virus in HKC monolayer cultures at temperatures between 37° and 30°C. The virus was found to propagate readily in the tissue at a temperature as low as 30°C, the lowest in this study. At this temperature the metabolic rate of the tissue culture cells decreased, CPE developed more slowly and the peak of virus multiplication also was retarded. Nevertheless, titer yields were higher than those formerly obtained at 37°C. Virus harvests of cellfree medium with reasonably high titers could be made repeatedly, within comparatively short intervals, from the same culture, beginning soon after CPE was observed and until CPE was almost complete. However, if the same culture medium was left in contact with the cells for 13 hr or longer the virus titers of the fluid harvests were relatively low. Lability of the virus, which could be expressed in terms of an 18-hr half-life at 30°c in this study, was believed to be the major factor responsible for this drop, and this, in turn, was believed to be enhanced by the increasing hydrogen ion concentration in the virus-cell culture especially at the vicinity of the surface of the tissue culture cell monolayer to which the newly released virus particles were suddenly exposed on release in a stationary tube. If the pH was not dropping and was within the practical range of variation of the tissue culture system, it had no detectable effect upon the half-life of the virus. However, a medium with a pH as low as 7.0 when first introduced into an active virus producing culture and left for a period of 12 hr was found to give a lower yield of virus than one at the pH of 7.3 and 7.6.
Demonstration of Multiple Hemagglutinating Factors in Sera of Ragweed-Allergic Individuals with the Help of Immunosorbents
Authors: Perelmutter L. , Freedman S. O. , Sehon A. H.
Published: January 2023

Skin-sensitizing and hemagglutinating antibodies were completely removed from sera of nontreated ragweed-sensitive individuals by absorption with adequate amounts of erythrocyte-WSR conjugates. However, when increasing volumes of these sera were added to a constant amount of immunosorbent, differences in the ability of skin-sensitizing and hemagglutinating antibodies to combine with the ragweed antigens on the immunosorbent were revealed. Thus, the immunosorbent was saturated with respect to skin-sensitizing antibodies before reaching its full capacity for the removal of hemagglutinins. Similar conclusions were derived from absorption experiments in which serum fractions, obtained by chromatography on DEAE-cellulose, were used. Moreover, it was shown that skin-sensitizing antibodies could be displaced from the immunosorbent by the addition of serum fractions containing only hemagglutinins.

From this study, it was also concluded that there exists a spectrum of hemagglutinating antibodies both in terms of their physicochemical properties, and in terms of their affinity for the antigens present on the immunosorbent. It was shown that even if skin-sensitizing antibodies have hemagglutinating ability, their contribution to the over-all hemagglutination titer is only minimal. Therefore, a simple relationship between hemagglutination and Prausnitz-Küstner titers cannot be expected.
Quantitative Studies on Reverse Cutaneous Anaphylaxis in Mice with Progressive Cryptococcosis
Authors: Abrahams Irving , Gilleran Thomas G. , Weiss Carl Broock
Published: January 2023

Intradermal injections of C. neoformans rabbit antibody specifically provoked cutaneous anaphylactic reactions in mice with active experimental cryptococcosis as well as in uninfected animals previously prepared by passive administration of purified cryptococcal polysaccharides. In general, passive anaphylaxis tests in mice with this fungal system were more sensitive (interms of Ab N required for reactions) than tests reported with other antigen-antibody systems and in guinea pigs, sensitivity was either the same or somewhat less than with other systems.

RCA tests became positive in mice after an initial lag period of 4 to 6 days following infection with C. neoformans and thereafter the intensity of the reactions, as measured by intradermal titrations with antibody, increased with time as the disease progressed. A direct correlation was demonstrated between fungus cell count in the brain and sensitivity of antemortem reverse cutaneous anaphylaxis (RCA) titrations. Absorption of C. neoformans antiserum with cryptococcal polysaccharide abolished its capacity to provoke reactions in infected mice. Intravenous injection of 0.1 mg Ab N resulted in severe or fatal anaphylaxis in mice that were infected 15 days prior to the test.

The possible application of the antiserum intradermal test to the study of human cryptococcosis is suggested.
Immunoelectrophoresis of Nasal Secretions Collected during a Common Cold: Observations Which Suggest a Mechanism of Seroimmunity in Viral Respiratory Infections
Authors: Anderson Truman O. , Riff Louise J. M. , Jackson George Gee
Published: January 2023

Human nasal secretions collected prior to and during various stages of naturally-acquired or experimentally-induced common colds were analyzed for the presence of serum proteins by the method of immunoelectrophoresis. These analyses showed that serum albumin was present in all nasal secretion.

Experimental challenge of volunteers by intranasal instillation of an infectious common cold secretion resulted in the appearance of serum gamma globulin in the nasal secretion within 48 hr in all subjects tested regardless of whether the individual developed a cold. Nasal washings from those volunteers who developed a symptomatic illness contained serum α- and β-globulins in addition to albumin and γ-globulin on the 4th and 6th postchallenge days. In contrast, nasal washings from asymptomatic volunteers showed only serum albumin and gamma globulin on the 4th and 6th postchallenge day. It is suggested that the latter group, those subjects clinically resistant to experimental challenge with the common cold virus, may exude specific antibody onto the mucous membrane within the first 48 hr after contact with the virus and by this mechanism abort further progression of the infection and the appearance of clinical illness.

The action of viral neuraminidase on serum proteins in whole human serum and nasal secretion was demonstrated.
The Function of Proteolytic Enzymes and Tannic Acid in Inducing Erythrocyte Agglutination
Authors: Pirofsky Bernard , Cordova Maria , Imel Theodore L.
Published: January 2023

The pretreatment of erythrocytes with bromelin, a proteolytic enzyme, results in a 2-to 3-fold increase in globulin coating the erythrocyte surface. This increased number of globulin molecules is not responsible for the agglutination of saline suspended erythrocytes by incomplete antibody. The pretreatment of erythrocytes with tannic acid increased the number of γ-globulin molecules coating erythrocytes. This increased number of globulin molecules is not responsible for the agglutination of γ-globulin-coated erythrocytes by antiglobulin serum. Neither proteolytic enzymes nor tannic acid function in hemagglutination procedures by inducing the coating of erythrocytes with a globulin molecule. Rather, by surface changes, they permit a normally nonagglutinating reaction to result in agglutination. It is postulated that the incomplete antibody has a potentially bivalent nature. The function of enzymatic treatment in converting incomplete antibodies to complete acting substances might well be the removal of steric obstructions resulting from the disposition of antigen and incomplete antibody sites.
Passive Anaphylaxis in Mice with Homologous Antiserum
Authors: Treadwell Perry E. , Rasmussen A. F.
Published: January 2023

Study of anaphylaxis in the mouse produced by passive and reverse passive sensitization was undertaken. Groups of mice were sensitized with several levels of homologous antibodies and shocked with 1, 8 or 100 times the equivalence dose of antigen at intervals from immediately to 1 week after sensitization. The severity of the resulting anaphylactic reactions depended upon the amount of antibody injected, the amount of antigen injected and the interval between the sensitizing and shocking injections. A biphasic response was observed in both passive and reverse passive anaphylaxis; maximal reactivity occurred 0 to 2 hr and 48 to 168 hr after sensitization with minimal reactivity from 6 to 24 hr after sensitization. The following conclusions were deduced from this study: a) the anaphylactic reactivity depends upon the amount of precipitable antibody, and b) upon the antigen-antibody ratio of the complexes formed in vivo as well as in vitro; c) the anaphylactic reactivity in the first 24 hr after sensitization depends primarily upon the formation of antigen-antibody complexes in the circulation, and after 24 hr, on an antigen-antibody reaction outside of the circulation.
Differentiation of Active and Passive Poliomyelitis Antibodies in Human Sera by Indirect Immunofluorescence
Authors: Riggs John L. , Brown Gordon C.
Published: January 2023

The indirect method of immunofluorescence has been shown to detect antibodies for poliomyelitis virus. The reaction is specific for immunologic type and the FA titers correspond to the neutralizing antibody titers but are consistently lower.

Paired sera from recovered cases and from vaccinated infants reflecting active antibody development show a marked increase in fluorescent antibody reaction. In contrast, most of the sera from infants containing high titers of passive antibodies which suppress active response to vaccine fail to stain by the indirect procedure of immunofluorescence.

Possible explanations for this phenomenon are discussed.
Effect of Molecular Weight on Uptake of Antigens by Immunologic Aggregates
Authors: Kahn Robert E. , Leskowitz Sidney , Lowell Francis C.
Published: January 2023

Washed specific precipitates were prepared from goat antiserum to rabbit γ-globulin and rabbit antibody to a series of protein antigens. The ability of these aggregates to bind antigen specifically was studied with I131-labeled materials.

It was found that aggregates prepared with rabbit antibody to ribonuclease, hen egg albumin, bovine serum albumin and human γ-globulin could, with sufficiently long incubation, take up approximately 80% of theoretic amounts of antigen. Aggregates prepared with rabbit antibody to hog thyroglobulin and busycon hemocyanin bound only small amounts of antigen.

The results suggest that the dimension of the lattice formed by goat antibody and rabbit γ-globulin permit antigens up to the size of human γ-globulin to enter and react with specific rabbit antibody sites. The larger antigens are almost totally excluded and react probably only at the surface of the aggregate. Each molecule of rabbit antibody is positioned in the aggregate so as to be capable of binding close to two molecules of antigen under optimal conditions.
Observations on Bacterial Anaphylaxis with Pneumococcus
Authors: Zinsser Hans , Mallory Tracy B.
Published: January 2023

Bacterial anaphylaxis was studied during the early periods of investigation into hypersusceptibility in general by Rosenau and Anderson and others who were pioneers in this subject. All of these workers, employing the methods of animal injection, were well aware of the difficulties of occasionally distinguishing between anaphylaxis-like reactions and true anaphylaxis. It is, of course, a well recognized fact, at the present time, that the frequent toxicity of bacterial extracts may produce symptoms not easily distinguished from those of the moderate anaphylactic reactions, and that the intravenous injection of any finely divided suspension may lead to phenomena practically indistinguishable from the specific reaction, by a mechanism not yet understood, but surely different from that of true anaphylaxis.

For this reason, one of the writers, with Parker, in 1917 employed the well known Dale method for the study of bacterial anaphylaxis, working with typhoid bacilli and their extracts (1).
On Antigenic Properties of Horse Serum and Egg Albumen After Heat Coagulation
Authors: Zinsser Hans
Published: January 2023

In a recent publication on the solubility of coagulated proteins,Wells and Lewis1 employed complement fixation and anaphylactic experiments to test the presence in solution of horse serum and egg white which had been completely coagulated by heat,and subsequently washed. Immunological reactions are of considerably greater delicacy than any known qualitative chemical test, and by these methods they succeeded in showing that after intraperitoneal injection into guinea-pigs, coagulated egg and serum proteins were dissolved in sufficient amounts to sensitize the animals to the homologous uneoagulated protein. They concluded from their experiments that heat coagulation is possibly a reversible process, and that such re-dissolved protein has lost none of its immunological antigenic characteristics.

In recent studies of our own, in which the question of heat coagulation and antigenic properties was involved, we encountered conditions which we believe have interest both in connection with the observation of Wells and Lewis, as well as in showing the considerable exposure to heat which proteins will withstand without being antigenically altered.
Pathogenesis of the Specific Inflammatory Reaction of Immunized Animals (Arthus Phenomenon)
Authors: Opie Eugene L.
Published: January 2023

Whenan animal has received repeated injections of foreign protein it reacts with acute inflammation often followed by necrosis whenthe antigen is reinjected into the skin. This reaction, usually designated the Arthus phenomenon, has been described asskin hypersensitiveness, local anaphylaxis, etc. It is not limited to the skin,for tissues of internalorgans,for example, the lungs or serous membranes, exhibit a similar heightened inflammatory reaction in consequence of repeated protein injections. The reaction makes its appearancewithin ten daysafter the injection of protein but doesnot reach maximum intensity until the animal has received six or eight injections at intervals of several days.No relation to anaphylactic shock has been established.

The Arthus phenomenon resembles very closely the tuberculin, luetin and typhoidin reactions which are produced when poducts derived from a microörganism are injected into an individual who has been invaded by the corresponding microörganism.
The Preparation of 0.1 L+ Toxin-Antitoxin Mixture
Authors: Banzhaf Edwin J.
Published: January 2023

The media for diphtheria toxin to be utilized for toxin-antitoxin mixture is prepared in the usual way with the following exception: To every pound of ground fat free veal a liter of water is added and placed in a refrigerator over night. The following morning the liquid is collected and boiled. The media is boiled at this stage to kill all organisms except the few heat resisting spores, thereby practically limiting the bacterial protein soluble substance to the coli and diphtheria bacilli which are later added. After boiling the reaction is corrected to 1.5 acid to phenolphthalein and 10 cc. of a twenty-four hour culture of B. coli is added for every liter of media and incubated about eighteen hours. One egg for every 2 liters is added, then boiled and strained. The reaction is corrected to 0.5 acid to phenolphthalein.
On the Susceptibility to Diphtheria (Schick Test Positive) with Reference to the Inheritance of Blood Groups
Authors: Hirszfeld Hanna , Hirszfeld L. , Brokman H.
Published: January 2023

By means of the Schick reaction, we were able to distinguish, among human beings, those susceptible to diphtheria from the immune. The blood of the individuals having a negative Schick reaction contains normal antitoxin, and these are resistant to the infection. The origin of normal antitoxin has not been heretofore explained. Most investigators regard them as representing an acquired immunity. However, we meet with Schick negative individuals who have not had diphtheria. In these cases, the authors suppose that we are dealing with the effect of unrevealed infections. The following data seem to support this opinion. The countrymen are more often susceptible than town dwellers. The poor are more often immune than the rich. Children are more susceptible than grown-ups, girls more than boys. The authors explain this greater immunity of the poor and boys by their being more exposed to infection.