bluuebunny/crossref_metadata_2025 huggingface dataset:
Hemolytic Properties of Eastern and Western Equine Encephalomyelitis Viruses
Authors: Karabatsos N.
Published: January 2023

Genetron-treated fluids of chick embryo cultures infected with Eastern and Western equine encephalomyelitis viruses were found, after freezing and thawing, to possess hemolytic activity over a wide temperature range (4 to 37°C). The virus hemolysin and hemagglutinin were inactivated by heat, whereas treatment with trypsin, ether and Ca++ tended to cause a selective suppression of the hemolysin activity of both viruses. Certain changes in the pH at which assays were performed resulted in a reduction of hemolytic activity which was greater than any change in hemagglutinating activity. Treatment of chick red blood cells with RDE and trypsin failed to remove the receptors for either of the activities. However, trypsin treatment significantly increased the hemagglutinating activity of both viruses. The hemolysins were inhibited by specific viral antibody and were sedimentable with the hemagglutinins.
A Study of Enzymatic and Antigenic Properties of Renin
Authors: Haas Erwin , Goldblatt Harry , Gipson Edwin C.
Published: January 2023

The procedure for the immunization of rats with renin was reinvestigated and optimal conditions for the formation of large amounts of antirenin within 3 weeks were established. This made possible the investigation of the effect on antigenicity of hog renin of treatment by enzymatic, chemical, immunochemical and physical procedures, known to exert profound effects on the structure of proteins, on the activity of other enzymes and on the immunologic specificity of antigens. These procedures included the digestion of renin by trypsin, papain, or pepsin, treatment with acid or alkali, exposure to carboxymethylation at various pH levels, reduction by various thiol compounds, depolymerization by urea, the detergent action of sodium dodecylsulfate, immunologic inactivation by antirenin, the blockade of functional groups by gold chloride and p-chloromercuribenzoate, nitroguanidination, and irradiation by x-rays or ultrasound.

Some of these treatments completely inactivated hog renin with respect to its enzymatic activity but spared to a variable extent the antigenic activity. Antirenin to unmodified hog renin is capable of inactivating the enzymatic activity of the renin of many animals but not of man. In no case was the antigenicity of modified hog renin so altered as to induce the formation of antirenin also capable of inactivating human renin.

Certain new properties of renin were established in the course of this study. It was shown that renin possesses an unusually high resistance to digestion by proteolytic enzymes. It can be exposed to reduction by cysteine without loss of activity, and subsequent exposure to atmospheric oxygen does not lead to its reoxidation. Exposure of renin to carboxymethylation in neutral solution results in complete inactivation of the pressor producing activity, but this activity can be protected by treatment with various thiol compounds. A reactive thiol group probably is not involved directly in the catalytic activity of renin. Renin which has been completely inactivated as a pressor agent by an excess of antirenin is still effective in the production of antirenin.

Various observations have been summarized in the proceedings of a recent symposium (6) which suggest that the suppression of antibody formation against antigen complexed with excess antibody is a general phenomenon. This does not apply to the renin-antirenin complex produced in vitro, for the product of this reaction remains antigenic, and retains the property of inducing the formation of antirenin. The choice of the species for immunization was found to be a determinant in the efficiency of the resulting antirenin to cross-react with renin of different species.

A distinction between enzymatically and antigenically active sites has become possible since complex formation, which abolishes the antigenicity of renin, retains its catalytic (pressor) activity (2), whereas various procedures which abolish the enzymatic activity of renin fail to destroy its ability to induce the formation of antirenin (Table V).
Mechanisms of Immunologic Paralysis by Pneumococcal Polysaccharide
Authors: Neeper Carolyn A. , Seastone C. V.
Published: January 2023

Immunologic paralysis by type I pneumococcal polysaccharide could be terminated with water-in-oil emulsion 14 days after its induction. Mice were not immunologically paralyzed by 500 µg of pneumococcal polysaccharide if water-in-oil emulsion was inoculated simultaneously, 2 hr after, or 2 days before. Resistance resulting from adjuvant alone was observed but was of a lower order of magnitude than that produced by adjuvant in paralyzed mice.

Large numbers of murine splenic and exudative cells failed to break paralysis. Paralyzed mice were made resistant to pneumococcal infection by injection of a suspension of normal splenic cells containing 0.5 µg of polysaccharide, but these results could not be regularly repeated.
The Pathogenesis of Lymphocytic Choriomeningitis in Mice: the Effects of Different Inoculation Routes and the Footpad Response
Authors: Hotchin John , Benson Lois
Published: January 2023

A detailed study of the different host responses after inoculation of mice with lymphocytic choriomeningitis (LCM) virus by different routes was made using s.c., i.p., i.c. and f.p. inoculations. Mice of two different strains were observed for clinical disease signs, the progress of which was followed objectively by recording the over-all weight changes of the mice for a 2-week period after inoculation; both aggressive and docile virus strains were used.

High doses of the docile virus strain given by the i.c. inoculation route did not regularly produce disease and death, particularly in the Swiss mice. This phenomenon was concluded to be due to immunologic paralysis. In this and in some other cases, the immunizing titer of virus preparations was higher than the LD50 titer.

Subcutaneous inoculation caused very low mortality with the production of immunity. Footpad inoculation production of immunity. Footpad inoculation produced a severe edematous response which was the most reliable and sensitive clinical method of detecting docile virus strains.

The measurements of weight changes in the inoculated mouse proved to be the most sensitive index of disease. By recording weight changes, it is possible to judge the end point of LCM virus titration after approximately 4 days instead of the usual 10 days to 2 weeks. The method also indicates the presence of virus in doses which ordinarily do not cause detectable clinical response. The findings are discussed in terms of the pathogenic mechanism of LCM infection.
Studies of Pasteurella Pestis
Authors: Elsler Daniel M. , Kubik Gayle M. , Von Metz Eleanor
Published: January 2023

A crude protein fraction (FIB) and a toxin were prepared from Pasteurella pestis grown in casein hydrolyzate mineral carbohydrate medium, by fractionating the culture filtrate with (NH4)2SO4. The protein nitrogen, sulfur, phosphorus and the ultraviolet light absorption values were similar to those published by other investigators. Electrophoresis, however, revealed two rather than one component. Both the FIB and the toxin were antigenic.

One inoculation of either protein 1/2 hr before subcutaneous infection with virulent organisms induced immediate protection against P. pestis but not against Pseudomonas pseudomallei or Pasteurella tularensis. The degree of protection induced by one inoculation of FIB increased as the interval between treatment and infection increased.

The immediate protection against P. pestis, induced by the proteins, was probably specific but the participation of nonspecific factors was not precluded.
Immune Cytolysis of Mouse Thymic Lymphocytes
Authors: Reif Arnold E.
Published: January 2023

Immune cytolysis was carried out by mixing a suspension of mouse thymic lymphocytes containing 97 to 99% viable cells with complement and various dilutions of rabbit antiserum or isoantiserum. Cellular integrity was determined after incubation for 2 hr at 37° C, by use of vital staining. An end point of 50% cell deformation was employed. As source of complement, hamster serum was almost nontoxic to mouse thymic lymphocytes. Two absorptions with mouse red cells and three absorptions with mouse liver removed the slight toxicity and any tendency for clumping. Guinea pig serum similarly absorbed was still slightly toxic. The count of deformed cells in the assay tubes containing antiserum changed relatively little between 1 and 3 hr after the start of incubation. This could be attributed to a balance between the rate of increase in the number of stained cells and their rate of disappearance due to complete lysis. Optimal conditions for assay of complement were determined. Complement titers determined with cells sensitized with varying concentrations of isoantiserum were relatively constant; in contrast, the complement requirement decreased with increasing concentrations of rabbit antiserum. This complement-sparing action of rabbit antiserum was only partially correlated with its complement-independent cytotoxicity. Optimal conditions for assay of cytolysin were determined. Direct proportionality between cytolytic titer and cell concentration was obtained over a range from approximately 5 to 20 million cells/ml. There was progressive loss of proportionality at lower cell concentrations. An unabsorbed rabbit antiserum to AKR thymic lymphocytes gave no evidence of specificity in its cytolysis of thymic lymphocytes taken from four different strains of inbred mice. The two isoantisera that were tested showed potent cytolysis of AKR thymic lymphocytes.
Studies on Delayed Hypersensitivity to Proteins and Protein Conjugates
Authors: Trakatellis Anthony C. , Stinebring Warren R. , Axelrod A. E.
Published: January 2023

Picrylation of γ-globulins suppresses both their ability to induce delayed hypersensitivity and anaphylaxis to the carrier protein in guinea pigs. Furthermore, picrylation decreases the reactivities of the test antigens in both systems. These data provide no evidence for a dissociation between the antigenic sites responsible for antibody production and delayed hypersensitivity.

Difficulties inherent in the application of chemically altered proteins to studies of this type have been emphasized.
Antigenic Relationship between Two Types of 7 S Guinea Pig γ-Globulins
Authors: Jeanette Thorbecke G. , Benacerraf Baruj , Ovary Zoltan
Published: January 2023

The antigenic relationship between isolated guinea pig 7 S γ1 and γ2 antibodies has been examined. The two γ-globulins cross-react when studied with rabbit antisera to whole guinea pig serum in immunoelectrophoresis and Ouchterlony plates. Absorption of rabbit antisera with the slow fragment of guinea pig γ-globulin, obtained by starch block electrophoresis of papain digest, removes this cross-reactivity. It is concluded that the known differences in biologic activities of these two types of γ-globulins must be due to different structures of the H chain.

Isolated guinea pig antibodies of various antihapten specificities show consistent differences in their immunoelectrophoretic patterns with respect to mobility and number of precipitation arcs observed. Some of the differences have been shown to reside in the slow fragment. However, no qualitative antigenic differences have thus far been shown by absorption studies of the rabbit antisera used.
Oral Immunization and Production of Coproantibody in Human Volunteers
Authors: Freter Rolf , Gangarosa Eugene J.
Published: January 2023

A group of nine human volunteers received daily doses of oral cholera vaccine for 4 weeks. At the end of this period, seven (78%) showed appreciable antibody titers in fluid stools and/or duodenal fluids. Coproantibody could be maintained in these volunteers for 12 weeks (i.e., the length of the experiment) by a single weekly oral dose of vaccine.

A primary course of daily oral doses for 2 (rather than 4) weeks was less effective in inducing coproantibody. Completely ineffective was a primary course of twice-weekly (rather than daily) oral doses for 4 weeks. A tentative schedule for induction of coproantibody in man by oral vaccine may therefore be formulated which consists of a primary course of one daily dose for 4 weeks, followed by one weekly dose for as long as coproantibody must be maintained. Oral vaccination is the only known method for maintaining coproantibody in man.

Coproantibody titers in the orally immunized volunteers were of the same order of magnitude as those found previously after parenteral vaccination, in spite of the fact that 75 times higher serum antibody titers were induced by parenteral vaccine. Consequently, coproantibody stimulated by oral immunization appears to be produced in the intestine rather than being derived from serum antibody.

Coproantibody titers in the duodenum of fasting volunteers were low, but increased sharply after a test meal of butter. Antibody titers in duodenal fluids were unrelated to the presence of bile. In many volunteers duodenal antibody titers were of the same order of mangitude as fecal titers, suggesting that release of antibody into the lumen occurs not only in the duodenum but also in lower portions of the intestinal tract. Presence of coproantibody in duodenal fluids was unrelated to the presence of bile pigments.
Passive Anaphylaxis in Mice with Homologous Antiserum
Authors: Treadwell Perry E. , Rasmussen A. F.
Published: January 2023

Various factors influencing the anaphylactic reactivity of mice have been studied. There is a direct relation between the amount of antigenantibody complex injected and the severity of the anaphylactic reaction. This complex must be present at a specific level within a limited period of time for death to occur. Mice surviving an initial reaction are desensitized to subsequent lethal reactions 10 min to 6 days later, the duration depending upon the challenging dose of complex. The Webster-Swiss (W-BRVS) strain is exquisitely sensitive to anaphylactic shock when compared to several other strains. Studies on desensitization with different types of anaphylaxis using different antigen-antibody systems for desensitization and challenge suggest that the depletion of a substance other than antibody or complement is responsible for the refractory state.
Direct Localization of Hemagglutinating Proteins on Paper Electrophoresis Strips
Authors: Miller John T. , Boyd William C.
Published: January 2023

A method for the direct demonstration by staining with erythrocyte suspensions of hemagglutinating proteins on paper electrophoresis strips is described.
The Induced Rejection of Guinea Pig Skin Homografts by a Partially Purified Transfer Factor
Authors: Powell A. E. , Ray Otis , Hubay C. A. , Holden W. D.
Published: January 2023

The effects of a partially purified transfer factor (PTF) on specific guinea pig skin homografts has been described. There is a significant increase in the rate of rejection of such grafts, compared with controls. PTF taken from nonspecifically stimulated animals is inactive, as is that prepared from second-set recipients. There is no obvious correlation between the size of the cutaneous transfer reaction and the anti-graft activity of these preparations.
Immunologic Studies in Urinary Tract Infections
Authors: Ehrenkranz N. Joel , Carter Mary Jo
Published: January 2023

Hemagglutinins (HA) to O antigens from bacteria in urine were measured in serial serum specimens from patients with urinary tract infections. Twofold or greater HA rises were found in 51 of 105 bacteria from patients with acute infections of varying severity.

HA rises to specific Escherichia, particularly strains possessing O antigens 4 and 6, were found primarily in patients with temperatures greater than 101° and in those with overt acute pyelonephritis. In these persons and in patients with chronic pyelonephritis high HA titers to heterologous E. coli O4 and O6 antigens were also found. Serologic and cultural findings of other Gram-negative bacteria were more randomly distributed. These serologic methods offer promise in documenting urinary tract infections and separating such severe infections as acute and chronic pyelonephritis from cystitis.
The Effect of X-Ray on the Antigenicity of Donor Cells in Transplantation Immunity
Authors: McKhann Charles F.
Published: January 2023

The x-ray sensitivity of normal histocompatibility antigens in mice was studied. Strong antigens associated with the H-2 locus were found to be resistant to as much as 12,800 r in vitro, in that mice inoculated with such heavily irradiated cells were immunized by them and gave second set responses to subsequent skin grafts. Weak antigens associated with H-1 and H-3 loci were effectively destroyed by as little as 400 r, failing to induce immunity after irradiation in vitro. The cells used to induce immunity were spleen cells and a methylcholanthrene-induced sarcoma. Possible biologic relationships between strong and weak transplantation antigens are discussed and a reason for their different sensitivities to irradiation suggested.
Agglutinins of the Red Kidney Bean (Phaseolus Vulgaris): A New Cytoagglutinin Distinct from Hemagglutinin
Authors: Tunis Marvin
Published: January 2023

Kidney bean extract has been investigated with respect to its human erythrocyte and nucleated cell agglutinating properties. In addition to the customarily found hemagglutinin, the extract contains two different agglutinin activities. These can be separated by differential absorption with red blood cell stroma and show activity against cells other than red blood cells. The first of these is associated with a fraction of bean extract containing the hemagglutinin and shows some physical properties in common with the hemagglutinin. The hemagglutinin and this nucleated cell agglutinin can be absorbed and recovered from stroma under the same conditions and both have the same temperature of inactivation. It is not yet known whether the two activities are a function of the same material or two separate agglutinins. The second nucleated cell agglutinin is found in another fraction not absorbable by stroma, is devoid of hemagglutinating activity, and requires a higher temperature than the aforementioned nucleated cell agglutinin to be inactivated.
Antigenicity of Lipid-Adsorbed Diphtheria and Tetanus Toxoids
Authors: Youngner J. S. , Axelrod Velma
Published: January 2023

Studies of the effectiveness of hexadecylamine (HDA)-adsorbed vaccines have been extended to bacterial as well as viral antigens. Adsorption experiments show that diphtheria and tetanus toxoids adsorb to HDA with great efficiency whereas about 30 to 40% of the protein, which is not toxoid, does not adsorb. A significant purification of toxoid therefore is achieved by adsorption to lipid.

Lipid-adsorbed toxoids produce an antibody response in guinea pigs equivalent to the response elicited with alum-precipitated antigen, indicating that HDA can act as an adjuvant in combination with bacterial as well as with viral antigens. Data are presented dealing with the influence of the interval between primary vaccination with adjuvant vaccine and secondary stimulation with fluid toxoid on antibody response.

In contrast to the failure of HDA to act as an adjuvant in combination with influenza virus when injected into rabbits, HDA does act as an adjuvant with diphtheria toxoid in this species. The results introduce a new factor into the study of lipid adjuvants, namely, the effectiveness of the adjuvant in different species as a function of the antigen used.
Biologic Properties of Polynucleotides
Authors: Yachnin Stanley
Published: January 2023

Polyriboguanylic acid synthesized by means of RNA polymerase has been tested for anticomplementary activity. 0.054 µmoles P poly G inhibits one 50% hemolytic unit of complement. Since poly GU is a more potent anticomplementary substance than poly IU of similar base composition, the lesser (1/7) potency of poly G as compared with poly I is thought to result from the fact that poly G exists in solution as a multi-stranded helix.
Immunologic Studies of Purified Viral Hemagglutinin
Authors: Tong Myron J. , Fong Jacob
Published: January 2023

A hemagglutinin fraction (HAF) of influenza A (PR8) virus, purified by sucrose density gradient centrifugation and shown to be highly homogenous by analytical ultracentrifugation, electron microscopy and a gel diffusion test, was used as an immunizing antigen in these studies.

Sera of guinea pigs immunized intraperitoneally with purified HAF contained antibodies which exhibited hemagglutination-inhibiting, virus neutralizing, and toxin neutralizing properties.

Mice immunized with purified HAF prepared from PR8 virus were resistant to challenge infection with the homologous strain of virus but susceptible to infection with a heterologous (FM1) strain of virus.

Guinea pigs immunized with whole virus and challenged intranasally with homologous virus exhibited more extensive and intensive histopathologic changes than nonimmunized animals; intranasal challenge of immunized animals with purified HAF duplicated some of the pathologic changes caused by challenge of immunized animals with whole virus.
Antibody Formation by Rabbit Lymph Node Cells
Authors: Attardi Giuseppe , Cohn Melvin , Horibata Kengo , Lennox Edwin S.
Published: January 2023

A microdrop experiment was done following a suggestion of Dr. Frank Lanni (1) that T5, having at least two antigens involved in its neutralization, might by itself serve to determine whether a single cell can make antibody against more than one antigenic determinant and to see whether the heterogeneity of anti-T5 antibodies has its origin in a corresponding cellular heterogeneity. Lanni had shown (2, 3) that the survival of T5 plaque-forming ability after exposure to antiserum differs depending on whether it is assayed on Escherichia coli B or F and that this difference is due to reactions of antibodies with two antigens on the bacteriophage. Thus by examining single cells of an animal immunized with T5 for antibody neutralizing T5 plated on B or on F one can see whether a single cell synthesizes both antibodies.

The materials and methods are as described in previous articles in this series (4–7).
Immunochemical Studies on Cross-Reactions of Antipneumococcal Sera
Authors: Goodman Joel W. , Kabat Elvin A.
Published: January 2023

A study of the cross-reactions of type XXII horse antipneumococcal serum with various dextrans has shown the presence of two populations of cross-reacting antibody molecules. The reaction of one of these with B512 dextran (N236) is inhibited by the isomaltose series of oligosaccharides, isomaltohexaose and isomaltoheptaose being equal on a molar basis and the best inhibitors. On absorption of these antibodies, a second population remains which cross-reacts best with dextrans with high proportions of 1,2-and 1,3-like linkages, and for which the best inhibitors are the disaccharides kojibiose, trehalose and isomaltose; the higher oligosaccharides of the isomaltose series were equal in potency to isomaltose on a molar basis.
Antigenicity of the M Proteins of Group A Hemolytic Streptococci
Authors: Fox Eugene N.
Published: January 2023

M proteins from four group A streptococcus serotypes have been extracted from cells with hot acid, further purified by column chromatography and analyzed serologically and chemically. The purified M proteins elicited typespecific antibodies when administered in various protocols to 95 rabbits in two or three dosages of 100 to 400 µg per intramuscular injection. These antibodies were assayed by the hemagglutination of tanned human erythrocytes sensitized with purified M proteins. Bordetella pertussis vaccine, sodium alginate, polyethylene glycol and dextran were used as adjuvants. Only the B. pertussis enhanced the M antibody response. Eighty per cent of the rabbits responded with positive titers ranging from 1:100 to over 1:25,000. An antiserum with a titer of 1:3200 passively protected mice against 800 LD50 doses of homologous live streptococci. Bactericidal tests with antisera also corroborated the effectiveness of the M protein immunization. None of the immunized rabbits exhibited any signs of anaphylactic reaction or delayed cutaneous hypersensitivity as a result of multiple inoculations of M proteins.
Studies on the Properties of Fragments of Guinea Pig γ1 and γ2 Antibodies Obtained by Papain Digestion and Mild Reduction
Authors: Nussenzweig Victor , Benacerraf Baruj
Published: January 2023

Specifically purified guinea pig anti-hapten antibodies were digested with papain according to the method described by Porter. Immunoelectrophoretic analysis of the digested γ1 and γ2 antibodies reveals three antigenically distinct fragments. The slowest migrating component at pH 8.2 (S fragment) is yielded by both antibody types and can be shown to combine specifically with the antigen. The other two components (F1 and F2) are obtained respectively from γ1 and γ2 antibodies and are responsible for the differences in mobility of the antibody type.

Upon dialysis of the papain digested antibodies, a crystalline fraction is obtained which can be identified as originating from the F1 component of the γ1-globulins. These crystals have the appearance of quadrangular plates and are barely soluble in neutral pH.

Attempts to inhibit passive cutaneous anaphylaxis in guinea pig skin with the whole papain digest or with a solution of F1 crystals failed, indicating that the structure present in γ1-globulin molecules and necessary for skin fixation is lost after proteolytic action.

Reduction and alkylation of guinea pig γ2-globulin in aqueous solution followed by acidification and filtration in Sephadex G-100 leads to the formation of two fractions. The first component A (75%) is heterogeneous and cannot be immunologically distinguished from the native γ2-globulin. The second, B, is contained in the papain S fragment, and contains only part of the antigenic determinants of the native γ2-globulin.
Isoantibodies to Human Cancer Cells in Cancer Patients Following Cancer Homotransplants
Authors: Itoh Tetsuo , Southam Chester M.
Published: January 2023

Homotransplants of human cancer cells were made in patients with cancer of various types and in various stages of progression. Tissue-cultured cancer cell lines (HEp 2, HEp 3 or RPMI 41) were used for 28 recipients, and cell suspensions prepared from surgical specimens were used for another 12. Homologous leukocytes or other non-neoplastic cells were admixed with some of the inocula. Sera collected at frequent but irregular intervals were tested for reaction with antigens prepared from HEp 2 cells, using the tanned erythrocyte agglutination technique.

All pretransplantation sera gave negative tests.

Twenty-three of the 28 recipients of tissue-cultured cells developed antibody at serum dilution titers of 1:10 to 1:40, usually by 14 days after transplantation. Five of the 12 direct homotransplant recipients developed antibody, but titers were only 1:5 or 1:10.

As previously reported, rejection of these cancer cell homografts in cancer patients was significantly delayed in about half of the recipients—as compared with rate of rejection in healthy recipients. There was no consistent relationship between homotransplant behavior and antibody response.

The role of serum antibody in homograft rejection, and the ability of cancer patients to produce serum antibody, are discussed briefly.
Adoptive Tolerance to Shigella Antigen in Irradiated Mice Receiving Spleen Cell Transplants from Unresponsive Donors
Authors: Friedman Herman
Published: January 2023

Spleen cell suspensions from young mice immunologically tolerant to Shigella paradysenteriae antigens, induced by neonatal injection of a large inoculum of the antigen, were not competent to form agglutinins when transferred to lethally x-irradiated recipient mice. Challenge injection of irradiated recipients with Shigella antigen at various time intervals following spleen cell transfer from Shigellatolerant donor mice did not result in normal levels of agglutinins, as occurred following transfer of spleen cells from normal or Shigella-immune donors. Irradiated mice receiving unresponsive spleen cells were capable of forming normal levels of antibody to sheep erythrocytes or bovine serum albumin. Adoptive tolerance could not be demonstrated with any degree of confidence in non-irradiated recipients capable of actively forming antibody to Shigella antigens, either with or without spleen cell transfer from normal, Shigella tolerant or Shigella immune donors.
An Accelerated Storage Test for Predicting the Stability of Suspensions of Measles Virus Dried by Sublimation In Vacuo
Authors: Greiff Donald , Rightsel Wilton A.
Published: January 2023

Several recent publications have described how the expiration dates of vitamin preparations stored at room temperatures may be predicted quantitatively on the results of short-term degradation studies at elevated temperatures (1–5). The fundamental studies of Garrett have been responsible for the greatest advances in this field (2). Briefly, the method is based on the observation that a suitable metameter of the potency (the potency itself or its logarithm) declines, at a selected temperature, linearly with time; further, that different rates of decline in potency (or its metameter) obtained at different temperatures are related to these temperatures and act in accordance with fundamental physical chemical principles (6).

Based on the foregoing, thermal degradation should follow the logarithmic form of the Arrhenius relation with respect to absolute temperature (T): log k = -(ΔHa/2.303R)(1/T) (1) where k is the specific rate of degradation of the biologic material, R is the gas constant and ΔHa is defined as the heat of activation.
Induction of Tolerance to Bovine Serum Albumin by Means of Whole-Body X-Radiation
Authors: Linscott William D. , Weigle William O.
Published: January 2023

Adult New Zealand rabbits given 500 or 550 r of x-radiation, followed 2 and 7 days later by intravenous injections totaling 300 mg of bovine serum albumin (BSA), were unable to make antibodies against this antigen when appropriately challenged 2 months later. The degree of immunologic unresponsiveness observed was related to the amount of BSA injected during the first week after irradiation: 1 mg reduced the subsequent antibody response after challenge by 70%, 12 mg reduced it by 95%, and 300 mg reduced it by more than 99%. The resulting unresponsive state was specific, in that tolerance to BSA did not occur following irradiation and injection of 300 mg of human γ-globulin. The unresponsive state was much less complete 4 months after irradiation and treatment with BSA than it was at 2 months. The amount of BSA per kg of body weight required to induce tolerance appears to be approximately the same for irradiated adult rabbits and for neonates.
II. Reverse Passive Cutaneous Anaphylaxis in the Guinea Pig with Ovalbumin
Authors: Liacopoulos P. , Liacopoulos-Briot M. , Halpern B. N. , Herlem G.
Published: January 2023

Reverse passive cutaneous anaphylaxis with ovalbumin and rabbit anti-ovalbumin antibody has been regularly produced in guinea pigs.

The quantity of antibody necessary to elicit such reactions was found to be the same as that required to sensitize guinea pig skin for direct anaphylaxis in a few minutes after the intravenous injection of antibody.

It may be concluded that in explanation of anaphylactic reactions in guinea pigs in the direct as well as the reverse procedure, since antibody “fixation” appears to be unnecessary in sensitizing guinea pig tissues, one can invoke an antigen-antibody system giving rise to complexes toxic for the guinea pig, providing some quantitative conditions have been respected.
Immune Degradation in Passively Sensitized Mice
Authors: Morrison Sherie Leaver , Terres Geronimo
Published: January 2023

The rate of antigen degradation in passively sensitized mice is dependent on the species of animal in which the antiserum was made. Using guinea pig antiserum, it was found that some of the antigen was degraded at an accelerated rate while some was retained and degraded at a slower rate than antigen alone. The retained antigen was complexed in circulation with guinea pig γ-globulin. Two different rabbit antisera were tested. Mice injected with one antiserum retained antigen; mice injected with the other antiserum degraded antigen at the control rate after an initial phase of accelerated degradation. With mouse antiserum antigen was always degraded at an accelerated rate. Various explanations of these results are discussed. The data suggest that retention and accelerated degradation are a consequence of the existence of two different types of antibody.
Studies in Borreliae
Authors: Felsenfeld Oscar , Decker Walter J. , Wohlhieter John A. , Rafyi A.
Published: January 2023

One strain of Borrelia turicatae and one of B. parkeri were studied. The organisms had chemical compositions similar to those of leptospirae and of Trypanosoma brucei. Three antigenic factors were isolated, of which two were proteins and one a protein or polypeptide complex. One antigen, designated B, was shared by B. turicatae and B. parkeri. It did not differ in strains isolated from the first attack and from relapse. Antigens A and C were strain and relapse specific. These two antigens also played a role in the immobilization and lysis of borreliae. They reacted with antibodies present in the β- and γ-globulin fractions of the immune sera. Relapse sera protected young rats against borreliae from both the relapse and the first attack, while sera collected after the first attack did not protect against borreliae isolated from the relapse. The immune sera had a strong immobilizing and lytic activity, the latter dependent on complement. The strains used in this experiment did not show adhesion. Lysis was, therefore, considered independent of the adhesion phenomenon.
Eaton Pleuropneumonia-Like Organism (Mycoplasma Pneumoniae) Complement-Fixing Antigen
Authors: Kenny George E. , Grayston J. Thomas
Published: January 2023

Some properties of a complement-fixing (CF) antigen for Eaton pleuropneumonia-like organism (PPLO) (Mycoplasma pneumoniae) are described. CF antigen was found to be heat stable and extractable with organic solvents. The properties of the antigen indicated that it either was lipid or was associated with lipid. Raw antigen was extremely anticomplementary (AC). AC activity of whole antigen could be reduced by boiling, while AC activity of solvent-soluble antigens was low. Antigen extracted by either chloroform or ether behaved as a complete antigen by eliciting antibody production in rabbits, while antigen produced by chloroformethanol-aqueous KCl partition (L antigen) was at best poorly immunogenic.

For diagnostic purposes L antigen was more specific and demonstrated larger antibody responses than either boiled antigen or ether-extracted antigen. Testing of 40 paired sera from Eaton PPLO isolation-positive patients revealed 36 with 4-fold antibody titer rises, while the 4 others had high antibody in the first sera obtained 7 or more days after onset of illness. Geometric mean titer of first sera was 1:2.3 and of second sera 1:48, indicating an average 20-fold antibody response. Only 12 of 364 (3%) other patients with similar acute respiratory disease who were negative for Eaton PPLO isolation showed 4-fold antibody titer rises with L antigen.
Inhibition of Antibody Production and Acquired Immunologic Tolerance by Actinomycin
Authors: Claman Henry N. , Bronsky Edwin A.
Published: January 2023

Actinomycin D delays the appearance of circulating antibody to Bentonite-bovine γ-globulin (BGG). Actinomycin D converts the tolerogenic property of soluble BGG to an immunogenic stimulus. This adjuvant effect of actinomycin is not seen with 6-mercaptopurine, 5-fluorouracil, nitrogen mustard or cyclophosphamide. The ability of actinomycin to inhibit the tolerance produced by soluble BGG is considered a) as a possible result of inhibition of a repressor of antibody production, b) as a possible result of an alteration in antigen processing.
Relationship between Affinity of Anti-Dinitrophenyl Antibodies and Their Biologic Activities
Authors: Hurlimann Jean , Ovary Zoltan
Published: January 2023

Affinity of rabbit and guinea pig anti-dinitrophenyl antibodies was measured by equilibrium dialysis and the use of DNPNS dye. Both methods gave concordant results.

Both antibodies were capable of sensitizing guinea pig skin for PCA reactions. Only antibodies of high affinity could give PCA reaction with the bivalent hapten, bis-DNP-lysine.

Both antibodies gave the same PCA reactions with DNP-BγG and DNP-RγG when these were highly derivated and in excess. With smaller amounts of these highly derivated antigens only the antibodies of higher affinities gave PCA reactions to the same extent, whereas the antibodies with lower affinities reacted only when four times more antibody was used.

With the weakly coupled DNP proteins, PCA reactions were provoked only with four times more antibody of lower affinity, and excess of antigen did not increase the threshold of PCA reaction.

No difference was observed in RPCA between antibodies of higher and lower affinities.

The significance of these results is discussed.
Rejection of Allogeneic Skin Grafts and Production of Isohemagglutinins by Sensitized Mice after Sublethal Irradiation
Authors: Tyan Marvin L. , Cole Leonard J.
Published: January 2023

Data are presented on hemagglutinin production and skin graft rejection by nonirradiated and irradiated mice sensitized by various means. Certain of these data suggest that hemagglutinin production and homograft sensitivity are mediated by separate products of cell differentiation, perhaps distinct cell populations.
Enterovirus Hemagglutination: Inhibition by Several Enzymes and Sugars
Authors: Lerner A. Martin , Bailey Elizabeth Jane , Tillotson James R.
Published: January 2023

Hemagglutination (HA) with the enteroviruses, echovirus types 3, 7, 11, 12 and 19 and coxsackievirus, group B, types 1 and 5, and with reovirus type 2 was studied. Pretreatment of suitable erythrocytes with β-glucosidase, maltase or hyaluronidase did not effect the later HA of these viruses. Trypsin treatments inhibited the red blood cells' later agglutination.

Pretreatment of all these viruses with β-glucosidase removed their HA property. Coxsackie-viruses group B, types 1 and 5, were similarly altered by incubations with hyaluronidase. Trypsin and maltase did not alter the HA of any of the viruses.

Inhibition of HA with these viruses was readily demonstrated with a number of sugars. The patterns and quantities of sugars necessary for these inhibition reactions were distinct for each virus. The mechanism of action of these sugars has not as yet been elucidated.

These experiments make it necessary to determine chemically if the capsids of enteroviruses and reoviruses are glycoproteins.
Immunizing and Toxic Properties of Particulate Antigens from Two Immunogenic Types of Pasteurella Multocida of Avian Origin
Authors: Heddleston Kenneth L. , Rebers Paul A. , Ritchie Alfred E.
Published: January 2023

Sac-like particulate antigens were isolated from avirulent cells of two immunogenically distinct strains of Pasteurella multocida by extraction with cold saline and centrifugation at 105,000 × G. They possessed many of the properties ascribed to endotoxins, i.e., they were toxic, high-molecular-weight, nitrogen-containing, phosphorylated lipopolysaccharides which were readily inactivated under mild acid conditions.

Almost 100% of the chickens given either intravenous injections of saline suspensions of the particulate antigens or subcutaneous injections of mineral oil emulsions of them were protected to challenges of live organisms which killed 100% of the controls. The same degree of protection was obtained in mice.

Rabbits injected i.d. with mineral oil emulsions of the particulate antigens gave strong specific antibody responses, but the degree of passive protection of mice by their antisera did not correlate with the amount of antibody. Good protection was obtained with antisera to one of the strains of P. multocida, but not to the other.

Injection of 0.1- to 0.5-mg amounts of the particulate antigens into mice, rabbits or chickens produced moderate to severe toxic effects such as depression and diarrhea; death frequently followed. Chickens showed many of the same signs after injection of fractional milligram amounts of the particulate antigen as have been observed with cases of acute fowl cholera.

Mice were actively and passively protected to the lethal effects of the X-73 antigen, but the degree of active protection was superior to the passive protection obtained with rabbit antiserum.
Heterogeneity of Antibody Producing Cells and Antibodies as a Result of Environmental Alterations in Tissue Culture
Authors: Patterson Roy , Suszko Irena M. , Orr Mary F.
Published: January 2023

The same spleen from chickens immunized in vivo and cultured in two types of media produced cells with different morphology. Those cultured in normal chicken serum had cells with the appearance of macrophages, large and small lymphocytes, plasma cells and monocytes. Cultures of the same spleen in egg white contained small cells with the appearance of either small lymphocytes or small plasma cells. The major immunoglobulin containing antibody in the chicken serum cultures and egg white cultures appeared to differ when studied by autoradiographic immunoelectrophoresis.
The in Vitro and in Vivo Effects of Cortisol on Interferon Production and Action
Authors: Mendelson Jack , Glasgow Lowell A.
Published: January 2023

Using the plaque reduction assay for interferon, it was found that cortisol inhibited inter feron production in infections with either an RNA (Chikungunya) or a DNA (vaccinia) virus in vitro. The depression of interferon production was not associated with an increased rate of virus replication. There was no effect of cortisol on the anti-viral activity of interferon in vitro. In vivo, the injection of cortisol 1 hr before an experimental viremia significantly reduced the amount of interferon in the sera of mice 4.5 to 6 hr after infection.
The Isolation and Characterization of a Ninth Component of Hemolytic Complement, C′3f
Authors: Inoue Kozo , Nelson Robert A.
Published: January 2023

Experiments are described which strongly indicate that nine fractions of guinea pig serum are required for the hemolytic action of C′. The new component, termed C′3f, reacts seventh in the hemolytic sequence, i.e., after C′3e but before C′3a, and produces an intermediate complex on the surface of sheep erythrocytes which is relatively stable. C′3f is difficult to separate completely from C′2 by column chromatography. Along with C′3d, potent C′3f reactivity is found in certain fractions of bovine plasma. A method is described for the measurement of C′3f in guinea pig, canine and human sera. It is depleted from the fluid phase by treatment with EAb C′1,4,2, 3c,3b,3e. About 70% of C′3f reactivity is lost during the absorption of serum with zymosan.

Initial studies indicate that C′3f is a macromolecule with a sedimentation constant of about 5.0. It has a half-life at 56°C of about 23 min. There is no major loss of C′3f reactivity when whole serum is treated with NH4OH, cobra venom, phlorizin or salicylaldoxime. Purified C′3f is rapidly inactivated by trypsin. The major portion of C′3f is precipitated from serum when (NH4)2-SO4 is added to yield a final concentration of about 2.2 M. As yet, there are no known co-factors or inhibitors of the hemolytic reactivity of C′3f.
Complement-Mediated Hemolysis in Agar Gel
Authors: Milgrom Felix , Loza Ulana
Published: January 2023

Complement and rabbit anti-sheep hemolysin diffusing against each other in sheep erythrocyte-containing agar gel form two zones of hemolysis which can be related to γG and γM hemolytic antibodies. Similarly, two hemolytic zones were observed in immunoelectrophoresis experiments in which the hemolysin was electrophoretically separated in an agar medium containing sheep erythrocytes and in which lysis was induced by diffusion of complement. In many experiments, the γG hemolytic zone was separated into two areas which indicated the presence of two molecular species of γG hemolytic antibodies. This finding was confirmed by the demonstration of two antigenic types of γG antibodies separated from sheep erythrocytes.

Anti-sheep hemolysins of rabbit and human origin formed circular zones of hemolysis when they were diffusing into agar containing sheep erythrocytes and complement. These zones could be reduced in size by allowing antisera with hetero- or isoantibodies for γG-globulins to diffuse against the hemolysins. The practical application of hemolysis in agar gel was explored in studying lysis of bovine erythrocytes by infectious mononucleosis sera and lysis of human erythrocytes by the hemolytic variety of human anti-A and anti-B antibodies.
The Proliferative State of Antigen-Sensitive Precursors of Hemolysin-Producing Cells, Determined by the Use of the Inhibitor, Vinblastine
Authors: Syeklocha D. , Siminovitch L. , Till J. E. , McCulloch E. A.
Published: January 2023

A study has been made of the effects of vinblastine, an inhibitor of cellular proliferation, on the number of hemolysin-producing cells which appear in the spleens of mice following injection of sheep red cells. Vinblastine was given either before, or after, administration of the antigen. When sheep red cells and vinblastine were given in close succession, doses of the inhibitor of greater than 0.1 mg per mouse were found to cause a decrease in the number of hemolysin-producing cells present in the spleen 3–4 days later. A dose of vinblastine of 0.4 mg per mouse produced a 5,000-10,000-fold reduction in the number of hemolysin-producing cells detectable at this time. In contrast, when doses of vinblastine up to 1 mg per mouse were given 3 days prior to injection of antigen, the decrease in numbers of hemolysin-producing cells after 3–4 days was no more than 2-fold, when tested using two different assay procedures.

The results obtained indicate that the precursor cells (antigen-sensitive cells) which respond to antigen by proliferating and giving rise to hemolysin-producing cells, are not proliferating as rapidly in the absence of antigen as in its presence. Thus, an important and early part of the response of these cells to antigen is a change in their state, so that they enter into a more rapid cell cycle.
Antibody Plaque-Forming Cells: Kinetics of Primary and Secondary Responses
Authors: Hege John S. , Cole Leonard J.
Published: January 2023

Numbers of Jerne plaque-forming cells per spleen, serum hemolysin concentrations, and the degree of resistance of the serum hemolysin to degradation by 2-mercaptoethanol have been determined in adult CBA and C3D2F1 mice as functions of time following primary or secondary intravenous sensitization with 4 × 108 sheep erythrocytes.

The characteristic secondary response yields fewer PFC and lower serum antibody concentrations than the primary response. When an adequate time interval (9 weeks or more) elapses between primary and secondary antigen injections, the magnitude of the secondary response begins to approach that of the primary.

Primary serum hemolysin is mercaptoethanol sensitive (19S immunoglobulin) while secondary serum hemolysin is largely mercaptoethanol resistant (7S immunoglobulin).

Generation times of 7 and 9 hr are obtained for plaque-forming cells during the first 4 days of the primary response.

A mathematical model is presented which relates numbers of antibody-forming cells to serum hemolysin concentrations. Using the model together with available experimental data, it is concluded that the great majority of the cells producing 7S hemolysin probably are not detected by the Jerne technique, as applied to the spleen.