bluuebunny/crossref_metadata_2025 huggingface dataset:
Antigenic Analysis of Influenza Viruses by Complement Fixation
Authors: Lief Florence S. , Henle Werner , Schrack William D.
Published: January 2023

The data presented indicate that the complement fixation test with homologous strain-specific viral (V) antigen is suitable for the evaluation of antibody responses following immunization with monovalent influenza A2 virus vaccines. The results obtained are compatible with those reported by others who used the hemagglutinationinhibition techniques for assay.

In two of the groups under study the incidence of specific anti-A2V responses was low after one or two intradermal injections of 20 CCA units of virus (18 and 60%), higher after one (63–73%) and best after two subcutaneous doses of 200 CCA units (τ;90%). When the first injection induced a specific antibody response, a second dose given within 2–4 weeks often failed to increase the titers, although negative reactors were thereby often converted to positive.

There was evidence that the incidence and height of anti-A2V responses following the first dose increased to some extent with advancing age of the vaccinees, but a second dose, as a rule, eradicated these differences.

A certain percentage of the subjects in all age groups who were given vaccine subcutaneously possessed measurable anti-A2V in their sera prior to immunization and prior to the dissemination of A2 virus. Their anti-A2V responses were higher than responses seen in persons lacking anti-A2V in the base line serum.

In the third study group, which had experienced an A1 influenza outbreak 8 months earlier, with the development in over 30% of the patients of anti-A2V along with antibodies to the infecting strain, even a single intradermal dose of 20 CCA units of A2 influenza vaccine yielded good anti-A2V levels in over 80% of the individuals. This unusually high incidence of responses to the small dose of vaccine was not restricted to those persons who had been overtly ill previously or had formed anti-A2V at that time. It is suggested, therefore, that a clinical or inapparent infection in the recent past with an apparently unrelated influenza A strain might have supplied a conditioning stimulus which resulted in the heightened responses to the A2 virus vaccine.

The implications of these findings have been discussed.
Effect of Trypsin on a Susceptible Strain of Influenza A Virus
Authors: Cleeland Roy , Sugg John Y.
Published: January 2023

The study dealt with a trypsin-sensitive line of influenza A virus. Incubation of this virus with 0.25% crude (Difco 1:250) or crystalline trypsin for 1 hr at 37°C completely destroyed the hemagglutinins and reduced the egg infectivity by more than 99%. A second line of virus derived from the same isolate as the trypsin-sensitive line and closely related to it antigenically, and several other strains of influenza A, showed no significant decrease in either hemagglutinins or infectivity when treated in a similar manner.

Crude trypsin inactivated the sensitive virus at a more rapid rate than did crystalline trypsin. The results presented indicate that the crude product contains some heat labile constituent(s) which alone is incapable of inactivating the virus but which does make it more readily available to trypsin action.

All preparations of the sensitive virus appeared to contain a small percentage of trypsinresistant particles, but subcultures of these particles in embryonated eggs always yielded virus populations composed primarily of trypsinsensitive virus, and in that respect they were indistinguishable from the original virus suspension. By employing special procedures, a stable line of virus that possessed greatly increased resistance to trypsin was obtained from the sensitive strain.

Repeated passage of the sensitive virus at either high or low dilution caused no significant variation in its susceptibility to trypsin, which suggests that this property might serve as an easily recognized and stable marker for genetic studies.
Studies on Organ Specificity
Authors: Shulman Sidney , Witebsky Ernest
Published: January 2023

By means of zone electrophoresis of human thyroiditis serum, the major protein groups have been isolated. Subsequent serologic testing by agglutination of tanned and coated erythrocytes has demonstrated that human thyroid auto-antibody belongs to the γ-globulin group of serum proteins. By specific absorption of thyroiditis serum followed by electrophoretic analysis of the absorbed serum, the identification of thyroid autoantibody with γ-globulin has been confirmed. Precipitation analysis indicates that thyroiditis serum can show either of two types of behavior. One type of curve is flat and broad and does not show inhibition in antibody excess; the other is tall and sharp and shows a considerable range of antibody-excess inhibition. Examination in the ultracentrifuge reveals that the human thyroid autoantibody is primarily a member of the 7 S family of serum globulins, as observed in two sera. These sera represented both kinds of precipitation characteristics.
Studies on Organ Specificity
Authors: Rose Noel R. , Metzgar Richard S. , Witebsky Ernest
Published: January 2023

The sera of rabbits immunized with pooled rabbit pancreas incorporated into Freund adjuvants contain antibodies that react with certain other rabbit pancreas extracts. They fail to react in vitro with the antibody-producing animal's own pancreas extract. No gross or microscopic evidence of damage to the pancreas was observed. The antibodies were therefore designated isoantibodies rather than auto-antibodies. These rabbit pancreas isoantibodies could be demonstrated by complement fixation, precipitation, and passive cutaneous anaphylaxis. The isoantibodies of the sera could be removed by absorption with some individual pancreas extracts but not with others. Absorption of the anti-rabbit pancreas rabbit sera with rabbit serum, erythrocytes, gastrointestinal and urogenital tract mucosae or other rabbit organs did not remove the rabbit pancreas isoantibodies. This finding was true even if the absorptions were carried out with organs from a rabbit whose pancreas extract provided a positive reaction. Absorption of these antisera with human A, B or O red blood cells or sheep red blood cells did not inhibit the homologous reaction with rabbit pancreas extract. The existence of at least three different rabbit pancreas isoantigens could be shown by gel diffusion precipitation experiments within the heterogenous strain of rabbits used in the study. Similar pancreas isoantigens have not been found so far in other experimental animals such as guinea pigs or dogs, even though these species contain the pancreas-specific antigens demonstrable by heteroimmunization of rabbits. The anti-rabbit pancreas rabbit sera did not cross-react with pancreas extracts of other species. “Natural antibodies” of the type described by Kidd and Friedewald could be domonstrated in these antisera both before and after immunization with rabbit pancreas extract, although an occasional increase in titer was observed during immunization. These antibodies could be differentiated from the pancreas isoantibodies by specific absorption and heat inactivation properties as well as by gel precipitation. The electrophoretic mobilities of the rabbit pancreas isoantigens were compared to that of human serum albumin in agar gel with the following results: one isoantigen had an electrophoretic mobility slightly greater than albumin, one isoantigen had an electrophoretic mobility similar to that of albumin, whereas the other had a lower electrophoretic mobility than human serum albumin. Immunoelectrophoresis studies showed the rabbit pancreas isoantibodies to be in the γ-globulin region.
Serologic Studies of the Neuraminidases of Vibrio Cholerae, Diplococcus Pneumoniae and Influenza Virus
Authors: Madoff Morton A. , Eylar Edwin H. , Weinstein Louis
Published: January 2023

Antisera were developed in rabbits to preparations of the neuraminidases of Vibrio cholerae and Diplococcus pneumoniae, and to influenza PR-8 virus. Neutralization of the neuraminidases of these organisms was studied by the methods of hemagglutination-inhibition, precipitin reaction, electrophoresis, and neuraminic acid release from α1-glycoprotein. Neutralization of bacterial and viral enzyme activity by homologous antisera followed typical enzyme-antibody neutralization patterns. No evidence of serologic cross-reactivity was found between either the two bacterial neuraminidases, or the bacterial and viral neuraminidases by any of the methods employed.
Mortality Patterns and Antibody Response in Chicks Inoculated with Eastern Equine Encephalomyelitis Virus
Authors: Byrne Robert J. , Robbins Mary Louise
Published: January 2023

The HI test was applied in detecting antibody in chickens experimentally infected with equine encephalomyelitis virus. Hemagglutination-inhibiting antibody is detectable at a high level prior to neutralizing antibody and persists for at least 140 weeks.

Resistance to disease from Eastern equine encephalomyelitis virus infection, demonstrated by diminishing mortality, increases in chickens between the 3rd and the 14th day of life. Infection in birds which are 14 days of age or older when infected can be detected by the rapid development of hemagglutination-inhibiting antibodies.
In Vivo Detection of Ferritin in the Normal Guinea Pig by Passive Cutaneous Anaphylaxis
Authors: Ovary Zoltan , Janoff Aaron
Published: January 2023

a) Intradermal injections of anti-guinea pig ferritin antiserum in large guinea pigs produced PCA reactions when dye alone was administered 3 hr later. b) These reactions were not inhibited by mixing the antiserum with transferrin but were neutralized after mixing antiserum and crystalline ferritin. c) The reactions cannot be attributed to interaction of γ-globulin with anti-γ-globulin antibody present in the antiserum as a contaminant. d) Nonspecific inflammatory substances in the antiserum have also been ruled out as participating in the observed reactions. e) It is concluded that physiologic titers of ferritin have been detected in the circulation of normal, large guinea pigs by the PCA reactions provoked by the intradermal injection of specific antiferritin antibody.
Studies on Pulmonary Alveolar Macrophages from the Normal Rabbit: A Technique to Procure Them in a High State of Purity
Authors: Myrvik Quentin N. , Leake Eva Soto , Fariss Bruce
Published: January 2023

A technique is described for procuring large numbers of alveolar macrophages in a high state of purity, uniformity and viability from normal rabbit lungs. The macrophages can be maintained in tissue culture and are phagocytic but multiplication has not been observed. The cells undergo marked changes in volume with little loss of viability when exposed to osmotic changes. The studies are interpreted as indicating that these alveolar macrophages comprise a constitutive flora of the normal rabbit lung.
Naturally Occurring Isoantibodies of the S Blood Group System in Cattle
Authors: Stone W. H. , Miller W. J.
Published: January 2023

Several normal sera with naturally occurring antibodies reactive with cattle cells possessing the subtype factors U1 and U2 are described and two representative ones are more fully characterized by absorption analyses. It was shown that these sera detect a new specificity called U′, possessed by U2 and not by U1 cells. The results indicate strongly that the antigenic specificity called U2 is qualitatively different from U1.

The new U′ reagents are useful in distinguishing cells containing U1 and U2 from those containing U1 only. The naturally occurring antibodies exhibit a marked prozone, which is a distinctive feature that may have contributed to the failure to detect this specificity before now.

A development of general interest is the illustration that differences in specificity between two reagents can not always be detected by their patterns of reactivity with even a large panel of known cells.
Studies on the Combining Sites of the Protein Antigen Silk Fibroin
Authors: Cebra John J.
Published: January 2023

For the study of its combining sites with antibody, silk fibroin was prepared by solution in lithium bromide, followed by dialysis. The fibroin had a molecular weight of 50,000–60,000 and contained only a slight amount of contaminants by immunologic assay. The number of antigenic sites occupied by antibody at equivalence varied with the rabbit immune serum but in most cases ranged from 2.6 to 3.2. The peptides in the soluble fraction following chymotryptic hydrolysis of fibroin were able to produce essentially complete inhibition of the fibroin-rabbit-antifibroin precipitin reaction at a concentration of 130 to 160 μm N per ml. Dialyzable peptides possessed differential inhibitory activity as shown by testing fractions obtained by partition column chromatography.
The Multiplication of Foot-and-Mouth Disease Virus in Trypsinized Calf Kidney and Tongue Cells and Its Use As Immunizing and Complement-Fixing Antigens
Authors: de Boer Carl J. , Bachrach Howard L.
Published: January 2023

Substitution of minced calf kidney tissue for bovine tongue epithelium in suspended cultures has not proved satisfactory for propagation of foot-and-mouth disease virus. Trypsin-dispersed calf kidney cell suspension, however, supported the multiplication of the virus to significant titers. The use of confluent monolayers of calf kidney cells in Povitsky flasks offers a very effective means to produce large quantities of virus for immunologic, biochemical and biophysical studies. Immunogenic studies with formalin-treated tissue culture virus in mice and guinea pigs indicate that it is capable of inducing antibody titers to significant levels. Acid inactivation of the virus is destructive to its immunogenic properties. Foot-and-mouth disease virus multiplying on monolayers forms a complement-fixing antigen.
Circulating Antibody and Anaphylaxis in Mice
Authors: Rothberg Richard , Talmage David W.
Published: January 2023

Using 1.0 mg BSA in incomplete Freund's adjuvant to sensitize mice, and measuring the antibody response as antigen binding per milliliter of sera with the ammonium sulfate salting out technique, experiments were conducted to provide additional information regarding the relationship of circulating antibody to anaphylaxis and the influence of genetic background on both anaphylaxis and the influence of genetic background on both anaphylaxis and the production of circulating antibody.

Evidence is presented to demonstrate that: a) C57 and DBA mice produced small amounts of circulating anti-BSA, and were resistant to anaphylaxis with BSA; b) CF1 mice produced more anti-BSA, and were highly sensitive to anaphylaxis; c) the severity of anaphylaxis observed in all three strains appeared related to the amount of antibody present; d) anaphylactic shock was not observed in any strain until circulating antibody could be demonstrated; e) a 2-mg immunizing dose of BSA resulted in an earlier antibody response and anaphylaxis in CF1 mice than did a 1.0-mg dose.
The Release of Histamine from Rabbit Platelets by Means of Antigen-Antibody Precipitates
Authors: Barbaro J. F.
Published: January 2023

Not all rabbit platelets and/or plasma possess the same capacity to release histamine when exposed to immune precipitates. The variability in ability to support histamine release can also be demonstrated with plasma and platelets from the same rabbit tested on different days. Precipitates prepared from rabbit antisera are more effective in histamine release than those from horse and sheep antisera. When the precipitate weight is above 200 μg of nitrogen, there is a direct relationship between composition of precipitate and histamine release. The greater the antibody-antigen ratio, the greater the histamine release. The capacity of an antigen-antibody system to release histamine does not correlate with its complement-fixing ability.
The Release of Histamine from Rabbit Platelets by Means of Antigen-Antibody Precipitates
Authors: Barbaro J. F.
Published: January 2023

The need for plasma in the release of histamine from rabbit platelets has been demonstrated and the necessity for plasma in the rabbit platelet system has been discussed. Although plasma is needed for histamine release by immune precipitates, diluted plasma has an enhanced releasing capacity. Plasma preincubated at temperatures ranging from 15–37°C for 30 min exhibited a decreased capacity to support histamine release by immune aggregates, whereas plasma preincubated at 44°C showed an increased capacity to support histamine release. Plasma heated at 56°C for 30 min still retains its capacity to release histamine, although less than unheated plasma. The thermolabile components of complement do not appear to be involved in histamine release. The possible role of C′3 and C′4 in the release of histamine are discussed.
Studies of Lymphocytic Choriomeningitis in Mice
Authors: Hotchin John , Weigand Heribert
Published: January 2023

The ability of LCM virus to cause disease has been studied in mice after inoculation at various time intervals following birth. Two different strains of virus and host were used and the results of the four combinations compared.

It was found that, in general, less disease and mortality occurred as the interval between birth and inoculation decreased, except when this interval was very short (2 hr) which caused high mortality. Some variations occurred with the different combinations of host and virus.

Mice inoculated during the first few days after birth frequently passed through a “runt” stage for several weeks, with severe retardation of growth and hair development. It was concluded that LCM disease is an immunologic conflict between the host and the viral antigen; this can end in any one of three ways: a) immunological tolerance or paralysis (persistent tolerated infection); b) death of the host; c) suppression of virus with immunity of the host.
Immune-Adherence with Leptospira Antigens
Authors: Linscott William D.
Published: January 2023

The immune-adherence (I-A) and microscopic agglutination (M-A) tests with living Leptospira antigens have been compared by parallel testing of hyperimmune and convalescent sera. Although there is good correlation between the two tests with regard to specificity, the I-A test is decidedly more sensitive. When excess complement could be used, allowing the attainment of maximal sensitivity, I-A titers were 50 to 500 times as high as the corresponding M-A titers. A serologic procedure of such great sensitivity may be of value in certain aspects of Leptospira research.

It has been shown that the sera of normal guinea pigs contain antibody-like substances which cause leptospirae to attach to human erythrocytes in an I-A reaction. This natural adherence activity is specific for particular serotypes and among random bred guinea pigs may vary considerably from one individual to another. Among inbred animals the variation observed appears to be reduced.
Nature of the Active Site of Antibenzoate Antibodies: Further Evidence for the Presence of Tyrosine
Authors: Pressman David , Nisonoff Alfred , Radzimski Gerald , Shaw Anita
Published: January 2023

The effect of variation of pH on the binding constant of anti-p-azobenzoate antibody with hapten indicates that there is an ionizable group in its site with a pK of about 10. This pH dependence of binding is consistent with the presence of a tyrosine, cysteine, lysine, or N terminal amino acid residue in the site. However, it has already been pointed out that an amino group (lysine or N terminal acid), if in the site, would have to be very resistant to acetylation compared to the other such groups in the antibody. The fact that the binding constant is the same at pH 6 as at 8 rules out the possiblity of histidine in the site.

The complete reversibility of the capacity to bind hapten after exposure of the antibody to pH 10 followed by restoration of neutral pH indicates that major irreversible structural changes in the antibody do not occur. Moreover, the total number of combining sites estimated by extrapolation of the binding curves to infinite free hapten concentration are the same within experimental error over the range of pH used.

Further evidence for the presence of a tyrosine in the site is the fact that the decrease in binding observed with extensive acetylation is reversible on exposure to high pH values. Such reversibility is consistent with the hydrolysis of an ester linkage rather than hydrolysis of an amide linkage and indicates the presence of tyrosine, serine or threonine or perhaps cysteine. The presence of tyrosine had previously been implicated through the effect of iodination on the antibody activity.

The reversal of acetylation is not complete. It may be that some irreversible alteration takes place in the molecule upon acetylation or that certain of the antibody molecules which have been acetylated are resistant to hydrolysis. This also suggests the possibility that the combining sites are heterogeneous with respect to chemical composition.
Serologic Studies of Myxovirus Para-Influenza 3 in Cattle and the Prevalence of Antibodies in Bovines
Authors: Abinanti F. R. , Hoerlein A. B. , Watson R. L. , Huebner R. J.
Published: January 2023

The relative sensitivities of the hemagglutination inhibition (HI), neutralization (N) and complement fixation (CF) tests to detect parainfluenza 3 antibodies in bovine sera were compared. The HI and N tests were essentially similar in sensitivity; the CF test was less sensitive but appeared to be useful in detecting recent infections. The temporal appearance and persistence of each type of antibody was studied.

A serologic survey of market cattle bled at slaughter showed that at least 70% had parainfluenza 3 antibodies and that infection had a wide geographic distribution. The data suggest that greater numbers of new infections occur during the fall than during other periods of the year.
A Comparison of the Growth Properties of Coxsackie Virus Strains A-9 and A-10 with Poliovirus Strain Mahoney in Cultures of Primary Human Amnion Cells
Authors: Dunnebacke T. H. , Mattern C. F. T.
Published: January 2023

Comparable cultures of primary human amnion cells were inoculated with poliovirus strain Mahoney, Coxsackie virus strain A-9 and Coxsackie virus strain A-10TC. The intracellular virus reached a maximal titer between 8 to 12 hr after inoculation for each of the three viruses. The release of virus into the medium followed its production in the cell by about 4 hr, but the maximal amount released by 30 hr after infection did not exceed more than one-fifth of the amount produced (cell associated, intracellular).

The cytopathic changes in primary human amnion cells which followed inoculation were identical for each of the three viruses. However, the temporal relationships between the production of these viruses and the appearance of cellular changes were markedly different. The early cytopathic changes preceded slightly the appearance of intracellular poliovirus. Only 30% of the cells were in early stages of cellular change when the maximal intracellular titers of Coxsackie virus A-9 were reached. At the time of the maximal production of Coxsackie A-10TC, less than 10% of the cells were damaged, although 90% eventually underwent characteristic cellular alterations and degeneration. The data for the Coxsackie viruses showed that the cellular changes occurred subsequent to virus formation.

The amount of virus produced in individual cells of comparable size was similar for poliovirus strain Mahoney and Coxsackie virus A-10TC, while 4 to 7 times that amount was produced after inoculation with Coxsackie virus A-9.
Plaque Formation with ECHO Virus Types 15 to 24
Authors: Maisel John , Moscovici Carlo
Published: January 2023

ECHO virus types 15 through 20 and types 22 through 24 were studied in Rhesus renal epithelial monolayers for plaque production and for morphology of the plaques formed. All the types studied produced plaques by the fifth day after virus seeding, and three different plaque patterns were observed. Small, irregular plaques with diffuse boundaries and hazy centers were formed by ECHO virus types 15 through 20 and by types 22 and 23; on the basis of plaque morphology, they apparently belong to the “Group A” of Melnick (11). ECHO type 19 also formed another type of plaque which was large, irregular, with diffuse boundaries and clear centers. This did not easily fit into either Group A or Group B. The third pattern was produced by ECHO virus type 24, and was samll, irregular, with sharp boundaries and clear centers; it might form a subgroup with types 1 and 13, and apparently belongs in Group A. To date, four distinct plaque patterns have been observed to be produced by ECHO viruses, and grouping of ECHO viruses according to plaque morphology should aid in their study.
Laboratory and Field Investigations of Bovine Myxovirus Parainfluenza 3 Virus and Vaccine
Authors: Hamparian V. V. , Washko F. V. , Ketler A. , Hilleman M. R.
Published: January 2023

A study was carried out to evaluate, in cattle, the efficacy of a killed Myxovirus parainfluenza 3 virus vaccine which was incorporated into an emulsified mineral oil adjuvant. The vaccine was highly active in eliciting neutralizing, HI and CF antibody following a single dose in bovines and in preventing infection following artificial challenge by the respiratory route. By contrast, a prior history of one or two artificially or naturally acquired infections with the virus did not prevent reinfection on challenge. Appraisal of the value of the vaccine in preventing the clinical epidemiologic entity “shipping fever” must await clarification of the etiology and host factors responsible for this syndrome and proof of efficacy under natural conditions in the field. Evidence was presented for natural infection with Myxovirus parainfluenza 3 virus among initially seronegative calves in isolation. The source of the infection was unknown but might have been of human origin or have resulted from activation of a latent infection. The hemagglutination inhibition test was more sensitive than the neutralization or CF techniques for detecting antibody against Myxovirus parainfluenza 3 virus.
Effect of Endotoxin on Antibody Production by Chicken Spleen Cells Transferred to Chick Chorioallantois
Authors: Sibal Louis R.
Published: January 2023

The effect of endotoxin on the antibody response to bovine serum albumin (BSA) by adult chicken spleen cells transferred to chick chorioallantoic membrane (CAM) has been studied under various conditions. Spleen cells from donor chickens given endotoxin either 24 hr prior to or in conjunction with antigen produced significant amounts of antibody on chick membranes 4 to 7 days after transfer. This process was demonstrable even when cells were taken as early as 1 hr after antigen injection of endotoxin treated donors. The antibody response by cells of donors given antigen and endotoxin simultaneously 48 hr prior to transfer appeared to be depressed. Spleen cells from normal chickens consistently produced antibodies on the chick membranes when removed 48 hr after antigenic stimulation of donors. Possible explanations for these observations have been discussed.
Prophylactic Effectiveness of Live and Killed Tularemia Vaccines
Authors: Eigelsbach Henry T. , Downs Cora M.
Published: January 2023

An attenuated USSR vaccine culture of P. tularensis was examined for dissociation and found heterogeneous with regard to colony type morphology. Live BV, the more immunogenic of two variants isolated from the culture, proved effective for the protection of the mouse against s.c. challenge with strain SCHU S4; in contrast, killed cultures were not immunogenic. Guinea pigs vaccinated with live BV cells and challenged with strain SCHU S4 exhibited an appreciably longer survival time than animals vaccinated with killed preparations but eventually they succumbed.

Although SCHU S1-11, one of several colony type variants of reduced virulence isolated from strain SCHU S1, proved equally as immunogenic for the mouse as BV and afforded the guinea pig greater protection than BV, the relatively high residual virulence of this variant for the guinea pig precluded its use as a vaccine strain. Serial animal passage of BV permitted the isolation of a culture that was ultimately selected for the production of live vaccine. The culture was designated the live vaccine strain (LVS) and is currently being used for the production of lyophilized live vaccine for use in man.

Strain SCHU S4 proved superior to strain 503 as a challenge strain for quantitating the protection afforded test animals administered live vaccines.

Data on the immunogenicity and storage stability of lyophilized live tularemia vaccine produced from cells harvested from GCHA and more recently from MCPH proved sufficiently encouraging to have warranted an extension of this study to a systematic evaluation of the vaccine in the monkey and subsequently in man.
Mouse Antibody
Authors: Anacker R. L. , Munoz J.
Published: January 2023

It has been shown that peritoneal fluid of mice immunized intraperitoneally with Ea in Freund's adjuvants may contain as much as 2 mg of Ab N/ml. Qualitatively, mouse peritoneal fluid appears to be very similar to mouse serum. Mouse antibody behaves like rabbit antibody in the quantitative precipitin test; insoluble precipitates are formed in extreme antibody excess, and soluble complexes are formed in extreme antigen excess. The anti-Ea antibody, the only one studied, was found only in the γ-globulin portion of the fluid. Various factors affecting the precipitin reaction such as incubation temperature, length of incubation and NaCl concentration have been investigated, and conditions which permit satisfactory and reproducible results have been established.
Bacterial Adherence: A Method for Detecting Antibody Production by Single Cells
Authors: Mäkelä O. , Nossal G. J. V.
Published: January 2023

A technique believed to be a further method for the detection of antibody production by single cells is described.

Rats were immunized with a variety of bacteria, and lymph node cells removed during the secondary response were studied for their capacity to induce motile bacteria to adhere to their surface. About 100,000 lymph node cells from immunized and nonimmunized animals have been studied, in drops containing either one cell or many thousands. It was found that adherence occurred only when the test bacteria shared an antigen with the immunogen. Ninety-eight per cent of the identifiable positive cells belonged to the plasma cell series. It was not possible to render plasma cells positive for adherence by incubating them in a high-titered immune serum either in vivo or in vitro.

Evidence is presented that the phenomenon depends on an antigen-antibody reaction between the bacteria and cell-associated antibody; that it is possible to differentiate two major types of adherence, viz., H-adherence and O-adherence; and that the positive cells actually are antibody-producers.

It is believed that this quick simple test may help in the screening of large cell populations for antibody-producing cells.
Antigenicity, Solubility and Chemical Stability of Renin Complexes
Authors: Haas Erwin , Goldblatt Harry
Published: January 2023

The ability of renin to function as an antigen for the formation of antirenin and the inhibition of the pressor activity of renin by the immune serum have been reconfirmed. This capacity of renin to stimulate the formation of antienzymatic antibodies can be impaired greatly by coupling of the renin to certain polymers of high molecular weight such as sodium desoxyribonucleate, hyaluronic acid or chondroitin sulfate.

The pressor activity of the renin was unimpaired when it was in the form of the complexes. This indicates that the enzymatically active groupings (centers) in the renin molecule were not affected by the complex formation.

Renin was investigated further in combination with five additional compounds known to form complexes with proteins, but all of these failed to interfere with the activity of renin both as a pressor agent and as an antigen in the formation of antirenin.

The solubility of renin was enhanced considerably by complex formation with hyaluronic acid, chondroitin sulfate and protamine.

Renin exposed to n-heptane was completely inactivated, but it was protected and retained its pressor activity in the form of its complexes with oleic acid or lecithin.
Antibodies to Denatured Desoxyribonucleic Acid in a Lupus Erythematosus Serum
Authors: Stollar David , Levine Lawrence
Published: January 2023

The reaction between the serum of a patient with disseminated lupus erythematosus and DNA has been studied by quantitative complement fixation. Thermally denatured DNA was shown to be a more reactive antigen than native DNA. Immunologic measurement of melting out and renaturation of the antigen characterized it as DNA. DNase-digested antigen preparations failed to react but were effective inhibitors. Lupus erythematosus (LE) serum detected serologic differences in denatured DNA preparations from various sources.
Studies in Experimental Eosinophilia
Authors: Litt Mortimer
Published: January 2023

Eosinophilia was induced in the peritoneal cavity of guinea pigs by the passive transfer of antigen-antibody complexes. These results were obtained in three systems: bovine serum albumin and rabbit antibody, hemocyanin and either guinea pig or rabbit antibody. Complexes were effective whether prepared in antigen excess, antibody excess or at equivalence. Activity disappeared from the antisera when the antibody was absorbed by the specific antigen. These experiments identify the active factor in serum as antibody and provide direct evidence that eosinophilia is induced by immune complexes. The conclusion is reached that eosinophilia is one of the consequences of antigen-antibody union and that eosinophils do not mediate the production of antibody. The possibility that a common mechanism underlies the eosinophilia which is noted in diverse pathologic states is discussed.
Typing of Echo Virus Isolates by Immune Serum Pools
Authors: Schmidt Nathalie J. , Guenther Raymond W. , Lennette Edwin H.
Published: January 2023

An “intersecting serum scheme” for the identification of viral isolates by testing against pools of type-specific ECHO virus immune sera is described. Each specific serum is included in two pools and identification is made by demonstrating neutralization of the agent by the two pools sharing a common immune serum. The identification of all ECHO virus isolates tested in this scheme agreed with that made in tests with individual immune sera. The ECHO virus immune serum pools did not neutralize isolates of the poliomyelitis and Coxsackie viruses.

A single cell culture tube may be used for each immune serum pool-virus mixture.
Some Immunologic Properties of Human and Dog Glomerular Basement Membranes
Authors: Steblay Raymond William , Lepper Mark H.
Published: January 2023

A consistently useful technique for producing human glomeruli and human glomerular basement membrane preparations has been demonstrated, based on the Krakower-Greenspon method. One human glomerular preparation was microscopically free of any capsular or tubular contaminants. Human and dog glomerular basement membrane preparations were found to produce CF antibodies in the rabbit. An in vitro cross reaction between human and dog glomerular basement membrane preparations was demonstrated by means of a CF test. The homologous titer was greater in all cases. The supernatant of the low speed centrifugate of the oscillated glomeruli, largely cellular, was found to be a good antigen in the in vitro CF test. It is not clear whether this is due to solubilized and fragmented basement membrane or shorn away cellular cytoplasm antigens or both. The most likely explanation for the above facts is that immunologically similar if not identical antigen(s) exist in dog and human glomerular basement membrane preparations of the purity herein described.
Studies of Mercaptoethanol-Dissociated Normal Human 19 S γGlobulin and Pathologic Macroglobulins from Patients with Macroglobulinemia
Authors: Reisner Catherine A. , Franklin Edward C.
Published: January 2023

Following reductive cleavage of normal 19S γ-globulin and four pathologic macroglobulins with mercaptoethanol, the dissociated fractions could be separated into three distinct components by chromatography on diethylaminoethyl (DEAE)-cellulose. Fractions 1 and 2 had s rates of approximately 8S whereas fraction 3 appeared to represent heterogeneous low molecular weight substances. The fragments shared certain antigenic groups with 7S γ-globulin. However, they differed from it in antigenic and electrophoretic properties, sedimented more rapidly in the ultracentrifuge and contained greater amounts of hexose. Fractions 1 and 2 generally differed from each other and the native macroglobulin in electrophoretic behavior and hexose content. The fragments of four of five proteins studied were antigenically deficient when compared to the native protein but could not be distinguished immunologically when compared to each other. The possible relationship of these structural units to the electrophoretic mobility of the native protein is discussed. The existence of several distinct 7 to 8S components, derived from the normal 19S γ-globulins, each of which differs from 7S γ-globulin, offers additional evidence for the existence of at least two major groups of immune globulins.
Influence of Magnesium and Calcium Ions on the Hemolytic Activity and Stability of Guinea Pig Complement
Authors: Fife Earl H. , Muschel Louis H.
Published: January 2023

Quantitative studies have shown that Mg++ and Ca++ each have specific roles in C′ activity, the Mg++ enhancing hemolytic activity, the Ca++ protecting the reagent from decay. Although addition of Mg++ alone significantly increased C′ activity, this was offset by a marked reduction of stability. Concomitant addition of Ca++, however, overcame the lability conferred by Mg++. The findings indicated that optimal concentrations of both cations in complement fixation would provide maximum hemolytic activity and stability of C′, and minimize possible adverse effects due to uncontrolled amounts of endogenous cations in test reagents. A relative excess of either cation inhibited the activity of the other. Thus, addition of one cation alone, particularly Mg++, to diagnostic tests seemed strongly contraindicated.
Complement-Fixing Antibody Response to Inactivated Poliovirus Vaccine
Authors: Lennette Edwin H. , Schmidt Nathalie J. , Magoffin Robert L.
Published: January 2023

The CF antibody responses induced by the administration of formalin-inactivated poliovirus vaccine (Salk type) were studied in two groups of 6- to 9-year-old children, totaling approximately 200 subjects, and a group of 47 army recruits, predominantly 18 to 22 years of age. Four-fold or greater rises in the CF antibody titer were elicited in 57% of the children with satisfactory pre- and postvaccination bleedings, but only half as often (28%) in the recruits. Detectable increases in CF antibody level were almost exclusively encountered in subjects (71 out of 74) who had had prior natural infection with one or more types of poliovirus as indicated by the presence of prevaccination neutralizing antibody. In these subjects, however, “heterotypic” CF antibody responses, i.e., to viral types for which there was no prevaccination neutralizing antibody, fairly often accompanied a homotypic response. In a few children bled 1 year after vaccination a marked decline in the CF antibody levels was evident but titers in the 1:4 to 1:16 range were still encountered more often than in the prevaccination blood specimen.
Hemorrhagic Encephalopathy in Chicken Embryos Infected with Influenza Virus
Authors: Grossberg Sidney E. , Hook Edward W. , Wagner Robert R.
Published: January 2023

Prior allantoic infection with homologous or heterologous influenza viruses, or Newcastle disease virus, protected chicken embryos against the hemorrhagic and lethal effects of neurotropic (NWS) influenza virus inoculated intravenously. The titers of NWS virus in brains of protected embryos were appreciably lower than those in fully susceptible controls, suggesting that the protective effect of prior allantoic infection was related to inhibition of NWS virus multiplication. The refractory state to intravenous challenge with NWS virus did not depend on the amount of interfering virus inoculated intra-allantoically but did require that active allantoic infection be established 8 to 16 hr prior to intravenous challenge. Intra-allantoic inoculation of myxovirus inactivated by heat or u.v. irradiation, or neutralized by immune serum, did not enhance resistance to NWS virus challenge. The small quantity of infectious virus disseminated from the allantoic site of infection to the tissues of the embryo seems insufficient to account for interference with NWS virus multiplication. However, multiple intravenous or intra-allantoic injections of infected allantoic fluid, freed of infectious influenza virus by centrifugation and heating, protected embryos against hemorrhagic encephalopathy.

These studies are consistent with the hypothesis that the protective effect of prior allantoic infection is related to in vivo production of an interferon that gains access to the embryonic circulation from the infected allantois.
The Serological Cross-Reaction between Bovine Serum Albumin and Anti-Ovalbumin
Authors: Weigle William O. , McConahey Patricia J.
Published: January 2023

BSA reacted with sera from both guinea pigs and rabbits immunized with five times crystallized OV, but OV did not react with the sera from rabbits immunized with crystalline BSA. The cross-reaction between BSA and anti-OV sera was the result of antibody produced against an impurity in the five times crystallized OV. This impurity could be detected by double diffusion in agar in a commercial preparation of OM, but not in preparations of OV, CA or lysozyme. Sheep albumin also reacted with anti-OV sera. Sera of rabbits and guinea pigs immunized with five times crystallized OV contained antibodies which reacted in agar with OV, CA and OM, but not with lysozyme. The reactions of egg white, OV, CA and OM with rabbit anti-OV also were analyzed by immunoelectrophoresis.
Studies on the Role of Leukocytes in Infection with Influenza Virus
Authors: Inglot Anna , Davenport Fred M.
Published: January 2023

A comparison was made of the associative reactions of PR8 strain of influenza virus with guinea pig red and white blood cells. Leukocytes adsorb virus more efficiently than do erythrocytes, but only a small proportion of the virus taken up by leukocytes was eluted. WBC-associated virus was less available to antibody than was RBC-associated virus, as judged by adsorption of antibody from solution and by neutralization of virus. The leukocytes of peritoneal exudates appeared to contain a higher content of virus than blood obtained from infected mice. Phagocytosis of virus-fluorescein labeled antibody complexes by leukocytes was clearly demonstrated but tissue culture cells showed only surface localization of antigen. The appearance of both types of cells treated with the hemagglutinin subunits of influenza virus obtained by extraction with ether was indistinguishable from cells treated with virus. The results of the studies with fluorescein labeled antibodies were obtained by the application of new techniques developed in the course of this investigation. The significance of the findings is discussed and the hypothesis is presented that leukocytes may serve to transport virus to extrapulmonary sites in infections of man and animals.
Transfer of Rabbit Lymph Node Cells to Neonatal Recipient Rabbits
Authors: Harris T. N. , Harris Susanna , Farber Miriam B.
Published: January 2023

When lymph node cells were obtained from adult rabbits, incubated in vitro with a filtrate of trypsin-treated Shigella, and transferred to irradiated neonatal recipient rabbits, agglutinins to Shigella appeared subsequently in the sera of these recipients. When approximately the same number of cells was given to neonatal and to adult recipients, the amount of antibody produced per transferred cell was estimated to be about one-fifth as much in the neonatal as the adult recipient. However, with decreasing numbers of cells transferred to the neonatal rabbits there was an increase in the highest relative values of antibody produced per cell until, when the numbers of cells were approximately proportional to the weights of the recipients, the maximal value for the amount of antibody per transferred cell in neonatal recipients was about half of the level in 1-kg recipients.

X-irradiation of the neonatal recipients prior to transfer led to higher antibody levels than those found in nonirradiated litter-mates, in the same range of ratios as that found in older recipients. Cells obtained from lymph nodes draining sites of injection of heterologous antigens did not lead to significantly different antibody titers than in the case of cells from uninjected donor rabbits.

Neonatal recipient rabbits previously injected with rabbit leukocytes, even on the first day of life, yielded lower agglutinin titers after lymph node cell transfer carried out 1 week later. Complete suppression of the antibody-inducing effect of transferred lymph node cells was found after the injection of 107 leukocytes, and decreasing degrees of suppression in the case of 106 and 105 leukocytes, respectively.
Studies on Helminth Immunity
Authors: Heyneman Donald
Published: January 2023

Summary and Conclusions

The time interval between initial infection with eggs of Hymenolepis nana and onset of a protective host reaction against a challenge with the same parasite has been restudied.

These findings confirm earlier reports of an extremely rapid onset of immunity to H. nana. It is concluded that a local, rapidly mobilized antibody reaction by the immediate cells affected could be responsible in spite of the brief period involved. If this can be proved immunologically, it would provide interesting evidence supporting the concept that locally concentrated cellular antibodies are an important host response to tissue invasion by helminths.
Studies on Organ Specificity
Authors: Rose Noel R. , Shulman Sidney , Witebsky Ernest
Published: January 2023

Hog thyroid crude extract and two fractions derived from it by ammonium sulfate precipitation—thyroglobulin and thyralbumin—were evaluated serologically, using rabbit antisera with various combinations of thyroid-specific and species-specific antibodies. Most of the organ specific thyroid antigen is precipitated as thyroglobulin between 1.60 and 1.70 M ammonium sulfate. Very little species-specific antigen precipitates in this fraction. Additional thyroid-specific antigens that do not cross-react with antiserum to heterologous beef thyroid extract are brought down by higher levels of ammonium sulfate. The thyralbumin fraction, precipitated between 2.50 and 3.00 M ammonium sulfate, contains little or no detectable thyroglobulin but reacts strongly with hog serum antiserum. Absorption of antithyralbumin rabbit serum with hog serum removes all of its demonstrable antibody content. In double diffusion gel precipitation tests, with thyralbumin antiserum, thyralbumin produces up to four lines, all identical with antigens of hog serum or hog kidney. By means of immunoelectrophoresis, thyralbumin can be seen to contain at least two components, migrating with mobilities of serum albumin and serum γ-globulin respectively. In the same experiment, thyroglobulin appears as a single or doubled are in the α-globulin region.
Immunochemical Studies of Elastase and Antielastase
Authors: McIvor Barbara C. , Moon Henry D.
Published: January 2023

In quantitative precipitin studies with elastase and antielastase serum, the maximum amount of precipitate developed in the region of slight antigen excess. With a greater excess of elastase, soluble complexes were formed. Studies on the inhibition of the enzymatic activity of elastase were conducted with antielastase γ-globulin using elastolysis as an index of enzymatic activity. The antigen-antibody precipitates were completely inert; the elastolytic activity of the supernatants was inversely proportional to the amount of antielastase γ-globulin used.