bluuebunny/crossref_metadata_2025 huggingface dataset:
Interferon in Rabbit Brain, Cerebrospinal Fluid and Serum Following Administration of Polyinosinic-Polycytidylic Acid
Authors: Cathala Francoise , Baron Samuel
Published: December 2022

The present study was undertaken to compare interferon levels in rabbit brain, cerebrospinal fluid and serum following parenteral administration of a synthetic, double-stranded RNA, polyinosinic-polycytidylic acid, a potent inducer of interferon. Following intravenous administration of polyinosinic-polycytidylic acid the titers of interferon in the brain and cerebrospinal fluid were always a small fraction of those in the serum. Intrathecal administration of polyinosinic-polycytidylic acid significantly increased the later-appearing interferon in the cerebrospinal fluid but did not significantly affect the interferon titers in the brain. Local injection of polyinosinic-polycytidylic acid intracerebrally did give rise to more sustained levels of interferon in the brain. However, the highest levels of brain interferon were always far below the highest levels which could be achieved in the serum. Local production of interferon in the brain and its meninges was indicated by the finding that after intracerebral administration of polyinosinic-polycytidylic acid the levels of interferon in the brain and cerebrospinal fluid exceeded those in the serum.
Enhancement of Virus Multiplication and Interferon Production by Cortisone in Ocular Herpesvirus Infection
Authors: Oh J. O.
Published: December 2022

Herpes simplex virus (HSV) was inoculated into the anterior chambers of rabbit eyes, and either saline or 25 mg of hydrocortisone was then injected daily into the subconjunctival tissues. The animals that received saline developed minimal corneal opacity in only 25% of their eyes; there was only brief multiplication of virus, but no interferon (IF) was detected in the ocular tissues. The cortisone-treated animals, however, showed much more severe corneal opacity in almost 80% of their eyes, enhanced and prolonged virus multiplication, and rather high titers of IF in the ocular tissues. The results suggest a) that the aggravation of corneal lesions of herpesvirus by cortisone treatment is not associated with reduced production of IF in the infected tissues, and b) that IF plays at best a very modest role in minimizing infection with HSV in the eyes treated with cortisone. A partial inhibitory effect of cortisone on anti-HSV action of IF was demonstrated in in vitro experiments with primary cultures of rabbit kidney cells.
Electron Microscopic Study of Erythrocyte Receptors with Labeled Antisera to Membrane Components
Authors: Howe C. , Spiele H. , Minio F. , Hsu K. C.
Published: December 2022

Hemoglobin-free membranes and erythrocytes of group O, after reaction with ferritin-labeled rabbit antibody to purified virus receptor substance, were examined by electron microscopy. Antibody was found to bind specifically to discrete sites, in thin sections of cells and ghosts, as well as in negative contrast preparations of washed membranes. The distribution of reactive sites was similar to that described by others for ferritin-labeled isoantibody. The same antibody labeled with fluorescein reacted with washed membranes to give a granular type of fluorescence. The relationship of the virus receptor substance-reactive sites to those containing blood group antigens is discussed.
Production of Lymphotoxin and Migration Inhibitory Factor by Established Human Lymphocytic Cell Lines
Authors: Granger G. A. , Moore G. E. , White J. G. , Matzinger P. , Sundsmo J. S. , Shupe S. , Kolb W. P. , Kramer J. , Glade P. R.
Published: December 2022

A number of continuous human lymphoid cell lines were examined for in vitro synthesis of the effector molecules, lymphotoxin (LT) and migration-inhibition factor (MIF). Twenty-six of a total of 39 lines established from human peripheral blood lymphocytes produced LT activity and 7 of 8 lines assayed secreted MIF. The amounts of LT elaborated varied considerably from one cell line to another and some lines apparently secreted none at all. The cell-toxin secreted in vitro by line 5287 displayed physical properties that were analogous to those of the cell-toxin secreted by phytohemagglutinin-activated normal human lymphocytes.
Repression of the Synthesis of Immunoglobulins in Newborn Mice by Antibodies Directed Against the Variable Region of Light Chains
Authors: Ruffilli A. , Compere A. , Baglioni C.
Published: December 2022

Antibodies directed against the variable half of human light chains have been shown to cross-react with some mouse light chains. Injection of these antibodies into newborn mice caused a repression of the synthesis of the cross-reacting light chains. This phenomenon is analogous to the allotypic suppression caused by antiallotypic antibodies. It indicates that antigenic determinants of the variable region of mouse immunoglobulin light chains are expressed in newborn animals.
Localization of Glycosphingolipids in Human Tissues by Immunofluorescence
Authors: Marcus Donald M. , Janis Rosamond
Published: December 2022

Two glycosphingolipids, ceramide lactose and globoside, were isolated from human erythrocyte stroma. Antisera to these compounds were prepared in rabbits and characterized by complement fixation and hapten inhibition. Immunofluorescence studies of human kidney revealed that globoside was present in the plasma membrane and cytoplasm of epithelial cells of the proximal convoluted tubule. In the spleen both glycolipids were detected only in the plasma membrane of reticuloendothelial cells of the red pulp. Although glycosphingolipids are found in many organs, these studies suggest that they are present in appreciable quantities in only a few types of cells. This observation provides the first evidence that glycosphingolipids may occur within cells as well as in the plasma membrane.
Comet Plaques: Evidence for Storage of Low Efficiency Hemolysins by Indirect Plaque-Forming Cells
Authors: Merchant Bruce
Published: December 2022

Although the plaques produced by direct “γM” plaque-forming cells (PFC) were originally thought to represent in vitro antibody synthesis (1), more recent findings suggest that most γM plaques result mainly from a respiration-dependent secretion of preformed high hemolytic efficiency antibody (2). Current observations now support the view that at least some indirect “γG” PFC contain sufficient stored antibody to account for much of the hemolysis ordinarily seen in the proximity of a single immune cell. In the course of experiments with hapten-specific indirect PFC frozen at a 1°C/min programmed rate (3,4), stored for 2 to 3 months in liquid nitrogen, and then thawed rapidly before plating, unusual comet-shaped streaks of hemolysis commonly appeared among the typical plaques.

Spleen cell suspensions were prepared as previously described (5) from adult New Zealand White rabbits immunized with 3 ml of 10% arsanilic acid- or sulfanilic acid-derivitized pigeon erythrocyte stromata on days 1, 3, 6, 17 and 130 and killed on day 134.
Metabolism of γG and γM Immunoglobulins in Normal and Hypogammaglobulinemic Chickens
Authors: Frommel D. , Perey D. Y. E. , Good R. A.
Published: December 2022

The half-lives of immunoglobulins G and M were studied in normal chickens, in chickens rendered hypogammaglobulinemic by early bursectomy and x-irradiation, and in dysgammaglobulinemic chickens that had been subjected to bursectomy only. The catabolic rate of labeled IgM was found to be independent of serum levels whereas the disappearance curve of labeled IgG was related to the serum concentration of this immunoglobulin. These findings parallel prior observations in humans having inherited or acquired immune deficiencies. In the chicken, the relationship of the half-lives of IgM and IgG was found to be 1:1.5 in contrast to that observed for mammals, in which a ratio of about 1:4 is characteristic. It can thus be concluded that the metabolic properties of the 7S immunoglobulins in chickens differ markedly from those of mammals.
The Effect of Heterologous Antilymphocyte Serum (ALS) on the Humoral Antibody Response to Salmonella Typhi “H” Antigen and Bovine Serum Albumin
Authors: Lance Eugene M.
Published: December 2022

The immunosuppressive effects of heterologous ALS were studied in mice immunized with a bacterial (Salmonella typhi “H”) and a protein (BSA) antigen. The principal conclusions are that depression of the primary response depends upon the administration of ALS prior to antigen, the magnitude of depression varies directly with ALS dose and inversely with antigen dose, the secondary response is little affected, and ALS pretreatment may set the stage for the induction of tolerance. These results are discussed in relation to current knowledge of the humoral immune response and the mode of action of ALS.
Studies on IgA
Authors: Anderson Truman O. , Zschocke Rainer H. , Bach Gerhard L.
Published: December 2022

Bromoacetyl cellulose was used to prepare a monospecific anti-IgA immunoabsorbent for the one-step isolation of IgA from small volumes of human serum. Two features were essential to this procedure. a) Anti-IgA globulin was freed of inert γ globulin by immunoabsorption and was used for coupling to bromoacetyl cellulose. The binding capacity of this immunoabsorbent was 15 mg IgA/g absorbent (using whole human serum). b) 3 M urea wash solution was used to minimize contamination of the eluted IgA by nonspecifically absorbed serum proteins. The IgA obtained by this method was 98.5% pure; it retained biologic activity and showed the electrophoretic pattern of IgA in serum.
Immunochemical Quantitation of Human C1 Esterase (C1s): Concentration in Normal Serum and Purification Yields
Authors: Nagaki K. , Stroud R. M.
Published: December 2022

Immunochemical quantitation of C1s is possible with the single radial immunodiffusion technique. C1s is the complement component with the lowest concentration for which this technique has been used. The normal adult concentration is 32.8 ± 6.2 µg/ml (1 S.D.). The C1s concentration in normal human serum is relatively insensitive to the method of storing and collecting blood samples. Sera or EDTA plasma can be kept at 0°C, -20°C or -70°C, so this should be a practical method for clinical laboratories. Clinical studies in various disease states are in progress. Using this method, the recovery and overall yield at each stage of purification as well as the specific hemolytic and esterase activities were calculated. The overall yield of C1s averaged 3% to 4% with specific hemolytic activities of 150,000 to 350,000 site-forming units/mg, and specific esterase activities ranging from 1500 to 2500 units/mg. These figures are the best obtained to date, but with increasing knowledge of this important serum enzyme more rigid criteria of purification are being determined with the goal of achieving high constant specific activity. The yields at various steps should be helpful in redesigning the methods of purification.
Delayed Hypersensitivity to Trichinella Spiralis
Authors: Kim C. W. , Jamuar M. P. , Hamilton L. D.
Published: December 2022

Guinea pigs were sensitized four times at 1-, 2- or 3-week intervals with crude saline extract of Trichinella spiralis larvae emulsified in Freund's complete adjuvant, then skin-tested 28, 56 or 84 days after the initial sensitization. The skin-test reaction was not typical of delayed hypersensitivity but was somewhat like the Arthus reaction. Electron microscopic examination of the lymphocytes from these animals showed structural changes that are generally associated with synthetic functions. In addition, passive cutaneous anaphylaxis (PCA) and precipitating antibodies were detected in their sera. The 2-week interval between sensitizations proved optimal for synthetic activity and for antibody production of both γ1 and γ2 classes. Correlation was noted between the degree of antigenic stimulus and proliferation of cellular organelles associated with protein synthesis in lymphocytes. Indeed, with the largest dose and optimal interval of antigenic stimulation, fine-structure studies indicated the production of lymphocytes very much like plasma cells. Lymph node cells from repeatedly sensitized donor animals transferred to normal recipients elicited negligible skin reactions, somewhat like the Arthus rather than the delayed hypersensitivity type. Even on day 1 after transfer of cells from repeatedly sensitized donors, antibody was detected by PCA in recipients, especially when the donors had been sensitized at 2-week intervals. Since PCA antibody was detected within 24 hr after transfer, it seems likely that the lymphocytes themselves produce antibody in recipients, at least until the lymphocytes possibly can develop into another type. Precipitating antibody was not detected in any of the recipients by either micro-immunoelectrophoresis or the ring test. In contrast, the transfer of delayed hypersensitivity was successful with lymphocytes from donors sensitized only once irrespective of dosage. The present findings suggest that animals repeatedly sensitized by the antigen apparently “lose” delayed hypersensitivity.
Studies on Human IgD
Authors: Saha Anil , Chowdhury Parimal , Sambury Sheldon , Behelak Youssef , Heiner Douglas C. , Rose Bram
Published: December 2022

Physicochemical studies were undertaken on two human IgD (Δ:λ) myeloma proteins. Sedimentation equilibrium analyses using a photoelectric ultraviolet scanner showed that the apparent weight-average molecular weight of IgD is 200,000 ± 2000. On the basis of carbohydrate content, IgD is a high carbohydrate immunoglobulin, the ratio of the oligosaccharide moieties being quite distinct from other high carbohydrate immunoglobulins, IgA, IgM and IgE. One IgD molecule differs considerably from another with regard to several amino acid residues, e.g., asp, glu, ser, ile, gly, ala and val. The proline content of IgD was low in comparison with the other immunoglobulins. One IgD myeloma patient excreted λ-type Bence Jones protein which contained 12 oligosaccharide moieties. Fluorescence emission maxima of IgD occurred at 326 nm and 330 nm, the excitation wavelengths being 278 nm and 295 nm, respectively. Temperature-dependent fluorescence emission spectra showed an absence of a definite transition temperature in the case of IgD, whereas both IgA and IgM showed a definite transition temperature of 60°C.
The Assessment by Immunofluorescence Methods of Humoral Anti-Myelin Antibodies in Man
Authors: Edgington Thomas S. , Dalessio Donald J.
Published: December 2022

The indirect immunofluorescence assay provides an immunologically specific primary binding test for the measurement and characterization of anti-myelin antibodies in man. Anti-myelin antibodies of the IgG class are found in 88% of normal individuals and appear to be acquired between 1 and 4 years of age, a period associated with active myelination of the central nervous system. A slightly higher titer of these antibodies is observed in the sera of patients with amyotrophic lateral sclerosis than in the sera of normal individuals and of patients with certain other diseases. Anti-myelin antibodies of the IgA and IgM classes are relatively infrequently detected in sera of normal individuals, although they appear with considerable frequency in sera of patients with amyotrophic lateral sclerosis and multiple sclerosis. No significant in vivo fixation of these antibodies could be detected by radioimmunochemical means, and it would appear that anti-myelin antibodies may reflect release of myelin immunogen rather than serve a primary role in the mediation of disease.
An Immunogenic Polysaccharide-Protein Conjugate
Authors: Fielder R. J. , Bishop C. T. , Grappel S. F. , Blank F.
Published: December 2022

A galactomannan isolated from the dermatophyte Epidermophyton floccosum was covalently linked to bovine γ, globulin by reaction with 2, 4,6-trichloro-s-triazine (cyanuric chloride). This polysaccharide-protein conjugate induced humoral antibody formation in the rabbit. The antibodies gave precipitin reactions with the polysaccharide, with the conjugate and with the 7 globulin. Absorption of the antiserum with γ globulin left. a preparation of antibodies that were specific for the polysaccharide. Hyperimmunized rabbits also developed cutaneous hypersensitivity to the polysaccharide.
Antigenic and Immunogenic Properties of Components Contained in Rabies Virus-Infected Tissue Culture Fluids
Authors: Schlumberger H. Dieter , Wiktor Tadeusz J. , Koprowski Hilary
Published: December 2022

Following differential centrifugation or zinc acetate precipitation, it was possible to fractionate rabies-infected tissue culture fluids into components which exhibited different complement-fixing, protective and immunogenic activities. The virion-depleted fraction showed higher complement-fixing activity than the fraction which contained essentially all the virions. The virion-depleted fraction obtained after precipitation of the virus with zinc acetate showed, in addition to complement-fixing activity, good protective activity. Thus, it is possible to produce material which contains a minimum quantity of infectious virus, is immunogenic and induces protection against challenge with virulent virus.
Role of the Thymus in Tolerance
Authors: Ellison George W. , Waksman Byron H.
Published: December 2022

Adult rats received 750 or 800 R of whole body x-irradiation with simultaneous thymic or splenic shielding and immediately thereafter were given intrathymic, intrasplenic, or intraperitoneal injections of suspensions of rat spinal cord or acid-extracted rat cord encephalitogen. Three weeks later the animals were challenged with the corresponding antigen in Freund's adjuvant. Animals receiving intrathymic encephalitogenic material showed a slight but definite decrease in the severity of experimental autoallergic encephalomyelitis (EAE). The acid-extracted encephalitogen showed four components after filtration through Sephadex G-200. Electrophoresis on 5% acrylamide gel in the presence of 8 M urea revealed at least four cathodally migrating and four anodally migrating bands.
Physicochemical and Antigenic Studies on Human γE in Respiratory Fluid
Authors: Newcomb Richard W. , Ishizaka Kimishige
Published: December 2022

The γE in sputum and nasal fluid specimens from asthmatic children behaves like the γE in serum on ion-exchange chromatography, gel filtration and ultracentrifugation. No secretory piece antigen was demonstrable on the γE in respiratory exocrine fluids. Like the γE in serum, sputum γE sensitized monkey skin for passive cutaneous anaphylaxis.
Immunosuppressive Effect of Serum from CBA Mice Made Tolerant by the Supernatant from Ultracentrifuged Bovine γ Globulin
Authors: Tong James L. , Boose Dorothy
Published: December 2022

In studies on tolerance to bovine γ globulin (BGG) induced in CBA mice by injection of the supernatant fraction of ultracentrifuged BGG, tolerance was not terminated by adoptive transfer of 108 spleen cells from normal or immune mice. Moreover, the serum of tolerant mice passively transferred to normal mice inhibited the immune response to BGG. This inhibition was attribute 1 to the presence of specific antibody in the serum of tolerant mice.
Mechanisms of Release of Constituents from Rabbit Platelets by Antigen-Antibody Complexes and Complement
Authors: Henson Peter M.
Published: December 2022

Two mechanisms have been described by which antigen-antibody complexes in the presence of complement induce release of vasoactive amines from rabbit platelets. The reaction of soluble antigen, antibody in excess, normal rabbit plasma and platelets caused adherence of the platelets to the immune complexes (which required the action of C3) and lysis of the platelets (which required the action of C6). The lysis was observed microscopically and was accompanied by release of vasoactive amines, nucleotides and cytoplasmic and lysosomal enzymes. The release did not require active participation of the platelet and was postulated to result from cell lysis by complement components activated by the immune complexes in or on the platelet membrane. A nonlytic, complement-dependent release of vasoactive amines resulted from immune adherence of platelets to erythrocytes or zymosan that had fixed complement components through C3. Vasoactive amines and nucleotides were released by the reaction, but not intracellular enzymes or potassium (measured with rubidium 86). Platelet energy and activated platelet esterases were required for the release, which appeared to result from the C3-induced adherence of the platelets to the particulate antigens.
The Unblocked N-Terminal Sequence of Chicken IgG λ-Like Light Chains
Authors: Kubo Ralph T. , Rosenblum Irwin Y. , Benedict Albert A.
Published: December 2022

The light chains of human and mouse immunoglobulins belong to two antigenically and chemically distinct classes, κ and λ (1, 2). Hood et al. (3) have suggested that the light chains of the immunoglobulins of other animal species can be classified as homologues of the κ and/or λ classes and that the ratios of κ to λ are variable among the species. The κ class has a free (unblocked) N-terminal amino acid and a C-terminal cysteine residue, whereas light chains of the λ class generally have a blocked N-terminal position, usually pyrrolid-2-one-5-carboxylic acid (PCA), and a penultimate cysteine residue at the C-terminal end (3, 4).

Light chains of both κ and λ classes have a unique structural feature: a variable region of about 107 amino acid residues in the N-terminal half of the light chain, and a common region of approximately residues 108 to 215.
Conversion of Non-Immune Rabbit Spleen Cells by Ribonucleic Acid of Lymphoid Cells from an Immunized Rabbit to Produce IgG Antibody of Foreign Light Chain Allotype
Authors: Bell Clara , Dray Sheldon
Published: December 2022

Spleen cells (SpC) from non-immunized rabbits were converted to antibody “indirect” plaque-forming cells (PFC) by incubating them with ribonucleic acid (RNA) extracts of lymph node cells (LNC) obtained from rabbits 18 to 24 days after they were immunized with several intravenous injections of 4 × 108 sheep red blood cells (SRBC). The RNA extract was inactivated by ribonuclease (RNAse) but not by deoxyribonuclease (DNAse) or trypsin, indicating that the conversion required intact RNA. “Indirect” PFC were developed with optimal amounts of goat anti-IgG-Fc fragment and these were not inhibited by 2-mercaptoethanol (2-ME) or goat anti-IgM but were inhibited by excess anti-IgG-Fc fragment. This indicated that the antibodies produced by the PFC are of the IgG immunoglobulin class. The conversion was specific for the SRBC antigen injected. The light chain allotype of the IgG antibody produced by the PFC was identified by use of anti-b4 and anti-b5 to develop the “indirect” PFC, to visualize the PFC by radioautography and to inhibit plaque formation. Most of the “indirect” PFC of the converted SpC (64% to 84%) possessed IgG antibody with the allotype characteristic of the donor of the RNA extract; some of the “indirect” PFC (4% to 32%) possessed IgG antibody with the allotype characteristic of the donor of SpC. When the “RNA-converted” SpC were lysed by freezing and thawing, the anti-SRBC titer of the lysate, measured by the localized hemolysis in gel technique, was 4096 compared to 4 for lysates from the same number of lysed, nonimmune SpC alone. Moreover, anti-SRBC antibody could not be detected in the RNA extracts. These results indicate that a component of the RNA extract, presumably RNA itself, is providing information for the synthesis of at least part of the IgG antibody molecule.
Independence of Allotypic Determinants and Antibody Specificity
Authors: Spring Susan B. , Nisonoff Alfred , Dray Sheldon
Published: December 2022

Studies were carried out to ascertain whether the specificity of an IgG antibody molecule influences the expression of the full complement of determinants characteristic of a given allotype. The presence of determinants was measured by the capacity of a given IgG preparation to displace from anti-allotypic antibodies all of a reference 125I-Fab fragment, possessing the same allotype, but derived from a different source. The results indicated that all IgG molecules studied, including specifically purified antibodies, possess a full complement of determinants (a1, a3, b4) and suggest that the portion of the surface of the molecule comprising the antibody-combining site is independent of the region of the surface which combines with anti-allotypic antibodies.
Detection of Immune Response Against Synthetic Polymers of Amino Acids Employing the Plaque-Forming Cell System
Authors: Egan Marianne , Maurer Paul H.
Published: December 2022

The secretion of anti-polypeptide immunoglobulins by popliteal lymph node cells from rabbits immunized with synthetic polymers of α-L and α-D amino acids was detected with the localized hemolysis in gel (LHG) assay employing polypeptidecoated tanned sheep erythrocytes. Plaque-forming cells (PFC) were detected 5 to 7 days following primary immunization of rabbits with poly(L-glu42-L-lys28-L-ala30)n, or poly (L-glu36-L-lys24-L-ala35-L-tyr5)n, and with poly (l-glu60-L-lys40)n, and poly (D-glu42-D-lys28-D-ala30)n, when complexed with methylated bovine serum albumin (MBSA). No PFC were detected 5 days after immunization with poly (L-glu42-L-lys28-D-ala30)n, poly (D-glu42-L-lys28-L-ala30)n or poly (D-glu42-D-lys28-D-ala30)n. Cross-reacting antibody was detected at the cellular level. The PFC were due to immunoglobulin secretion since they were inhibited by sheep anti-rabbit globulin and specific anti-allotype serum.
Measurement of IgA Levels in Human Cord Serum by a New Radioimmunoassay
Authors: Faulkner W. , Borella L.
Published: December 2022

A new quantitative inhibition immunoassay for measurement of human IgA has been developed, based on the Single Antibody Millipore Filtration (SAMF) method of Borella. IgA labeled with 125I was mixed with anti-human IgA serum, incubated for 24 hr at 37°C and filtered through a 220-mµ pore size Millipore filter. Labeled antigen-antibody complexes were retained by the filter while unbound 125I IgA was filtered through. The concentration of IgA specifically bound to antibody was calculated by the radioactivity retained in the filter. Addition of nonlabeled IgA to the reaction mixture inhibited the binding of 125I IgA by antibody. Between 20% and 80%, this inhibition was a linear function of the concentration of nonlabeled IgA. This immunoassay was used to measure the concentration of IgA in cord serum and all of 60 specimens had measurable levels of IgA. The mean value was 0.8 mg%, ranging from 0.15 mg% to 2.15 mg%. The SAMF immunoassay is rapid, reproducible and highly sensitive and should provide an ideal method for future studies on the ontogenetic development of IgA and its relationship to perinatal infections.
Relationship of Epstein-Barr Virus-Induced Membrane Antigens in Lymphoid Cells to Viral Envelop Antigens
Authors: Gerber Paul , Goldstein Gerald
Published: December 2022

Immunofluorescent membrane antigens of Epstein-Barr virus-infected lymphoid cells from a normal donor and from a patient with Burkitt's lymphoma appear to be identical and to contain components of the viral envelop.
A Pre-Albumin Activator of Prekallikrein
Authors: Kaplan Allen P. , Austen K. Frank
Published: December 2022

Chromatography of human serum on diethylaminoethyl cellulose reveals five peaks having bradykinin-generating activity. Kallikrein is found in peak 1, Hageman factor is found overlapping peaks 2 and 3, and a prekallikrein activator is found in peaks 2 through 5. Further purification of the prekallikrein activator by gel filtration, carboxymethyl cellulose chromatography, and elution from unstained sliced disc gels after electrophoresis gave purified material which appeared heterogeneous and had an estimated molecular weight of 30,000 to 40,000. Throughout each step of purification, the prekallikrein activator retained the ability to correct specifically the coagulation defect of Hageman factor deficiency. Prolonged dialysis of a partially purified preparation of active Hageman factor resulted in the development of pre-albumin bands in the position previously noted for the highly purified prekallikrein activator. The development of these bands and their associated clot-promoting and prekallikrein-activating abilities at low ionic strength and the prevention of these events at high ionic strength but under otherwise identical conditions are consistent with the direct derivation of the low molecular weight pre-albumin prekallikrein activator from active Hageman factor.
Persistent Lymphocytic Choriomeningitis Viral Infection
Authors: Oldstone Michael B. A. , Dixon Frank J.
Published: December 2022

Mice naturally infected transplacentally with lymphocytic choriomeningitis virus carry high titers of virus in their blood and tissues, make anti-viral antibody, and develop an associated chronic disease. The virus travels in the circulation in virus-IgG-complement(C) complexes. A study of the renal glomeruli of such transplacentally infected mice showed the presence of viral antigen, C3 and Ig at birth, and a progressive accumulation of virus-Ig complexes that led to death from glomerulonephritis in some mice by 2 weeks of age.
Studies on the Immune Capabilities of Chiroptera
Authors: Hatten Betty A. , Allen Rae , Sulkin S. Edward
Published: December 2022

K values based on 90% neutralization of φX174 indicate that the response of bats held at 24°C and 37°C to immunization with multiple doses of 1010 plaque-forming units (PFU) was less and that production of ME-sensitive antibodies was more pronounced than in rabbits or guinea pigs. Neutralization of φX174 by early bat antibodies was especially dependent upon the presence of a heat-labile serum factor or factors. Once a response was elicited in bats (24°C), antibody production continued at a relatively high level for a long period of time. An anamnestic response was demonstrated at the latter time interval in which the quantity of predominantly ME-resistant antibodies, having minimal requirements for heat-labile factors, approached that induced earlier in guinea pigs. A rapid response with early conversion to ME-resistant antibodies was evoked in bats (24°C) after single doses of 1011 PFU of φX174 (without adjuvant) or 109 PFU of φX174 in complete adjuvant. Considerable dissociation of φX174 from bat antibodies occurred when less than 90% of the phage had been neutralized, but was not remarkable when inactivation was greater than 90%. The proportionate loss of anti-φX174 activity in relation to concentration of antibodies diminished upon dilution of bat plasma pools as the immune response progressed, suggesting that an increase in affinity of antibodies for φX174 had occurred, possibly in association with changes in the major type of antibody being produced. Potential effects of the nature and degree of antigenic stimulation upon immune responses in bats are discussed.
Degradation of Immunoglobulins by Intracellular Proteases in the Range of Neutral pH
Authors: LoSpalluto Joseph J. , Fehr Kurt , Ziff Morris
Published: December 2022

A proteolytic enzyme active in the range of neutral or slightly alkaline pH has been partially purified from a lysosome-containing granule fraction of human spleen. The molecular weight of the enzyme was estimated to be in the range of 15,000 to 30,000 by gel filtration, and it has a pH optimum of approximately 7.5 with bovine hemoglobin or human IgG as substrates. Its activity is stimulated by cysteine but is strikingly inhibited by diisopropylfluorophosphate. The enzyme is active against a variety of protein substrates including human IgM, IgG, IgA, albumin, and also bovine hemoglobin. The products of IgG and IgM digestion were studied most extensively. IgG was broken down into Fab and Fc fragments, which proved to be very similar in size and immunochemical properties to fragments produced by papain digestion. In addition, a slightly degraded product with a molecular weight in the range of 110,000 to 120,000 was observed, which was not distinguishable from intact IgG by immunodiffusion analysis. IgM was extensively degraded by the neutral protease, a large proportion being broken down to small, dialyzable fragments. Relatively small quantities of large fragments containing Fab and Fc(μ) determinants were found. Evidence was obtained that both Fab and Fc regions of the IgM molecule are degraded by this enzyme.
A Cytolytic System for the in Vitro Detection of Cell-Mediated Immunity to Soluble Antigen
Authors: Henney Christopher S.
Published: December 2022

An in vitro cytotoxic test is described for the demonstration of cell-mediated immunity to soluble antigen. This system has been used to study the development of a cytotoxic splenic lymphocyte population in the guinea pig after immunization with dinitrophenylated human IgG (DNP-HGG). As “target” cells, 51Cr-labeled mouse mastocytoma cells were used, and were coupled to DNP-HGG with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide HCl. “Target” cell destruction was assessed by comparing the rates of 51Cr release from these cells in the presence of splenic lymphocytes from specifically sensitized animals with that released in the presence of the same number of lymphocytes from unimmunized animals. The mastocytoma cells were found to be preferable to erythrocytes as carriers of antigen, in that they were more sensitive to membrane permeability changes. Splenic lymphocytes from guinea pigs immunized with DNP-HGG caused specific lysis of DNP-HGG-coated mastocytoma cells. Cells coated with DNP-BSA and HGG were lysed to a lesser extent, while cells coated with antigens immunochemically unrelated to the antigen used for sensitization were not lysed to any greater extent than that observed with lymphocytes from unimmunized animals. Several factors distinguished cytolysis by “sensitized” lymphocytes from that mediated by serum antibody in the presence of complement. Cytolysis by lymphocytes was not inhibited by anti-guinea pig immunoglobulin antisera. The total cytolytic activity of splenic lymphocytes did not parallel measured serum antibody. The specificity of cytolysis by splenic lymphocytes from animals immunized with DNP-HGG was markedly carrier-dependent.
Release of Virus Inhibitor from Tuberculin-Sensitized Peritoneal Cells Stimulated by Antigen
Authors: Milstone Leonard M. , Waksman Byron H.
Published: December 2022

Tuberculin-sensitized mouse peritoneal cells exposed to purified protein derivative release an interferon-like virus inhibition within 24 hr. Release appears to require the interaction of specifically sensitized lymphocytes and sensitized or unsensitized macrophages.
Heterogeneity of Blastogenic Factors Produced in Vitro by Antigenically Stimulated and Unstimulated Leukocytes
Authors: Kasakura Shinpei
Published: December 2022

Blastogenic factor derived from antigenically stimulated or unstimulated leukocyte cultures was studied. Cell-free medium (CFM) from unmixed leukocyte cultures was frequently stimulatory for both autologous and allogeneic cells. However, the blastogenic factor usually stimulated allogeneic cells much more than autologous cells. The blastogenic factor which is stimulatory only for allogeneic cells may be a soluble form of histocompatibility antigen. The CFM preparations that were the most stimulatory toward allogeneic cells were not necessarily the most active toward autologous cells. The two types of stimulatory activity showed no correlation. It seems probable that CFM from unmixed cultures of leukocytes contains at least two components: a) a specific blastogenic factor (histocompatibility antigen), and b) a nonspecific stimulatory factor (probably identical to the “potentiating factor” described by Janis and Bach). Blastogenic factor was produced in vitro by tuberculin-sensitive leukocytes in response to PPD-stimulation. The production of this factor was antigenically-specific, but once produced, it was as active for autologous cells as for allogeneic cells. This factor is not histocompatibility antigen. The continuous presence of antigen was not required for activity. This factor may be different from the “lymphocyte-activating factor” described by Valentine and Lawrence. When the medium was changed every 2 days, the blastogenic factor was produced repeatedly until the culture died. The present findings indicate that the blastogenic factor produced by PPD-stimulation differs from that which is produced in unmixed cultures and which is stimulatory only for allogeneic cells. However, it is not known whether there are differences between the blastogenic factor arising from PPD-stimulation and the factor which is produced in unmixed cultures and which stimulates autologous cells.
Production of Congenital Malformations Using Tissue Antisera
Authors: Bragonier J. Robert , Frank Michael M. , Brent Robert L.
Published: December 2022

Studies have been performed in a model teratologic system in which congenital malformations are produced in offspring by injecting sheep anti-rat kidney antiserum into pregnant rats on day 8 of gestation. To determine whether the antiserum produces its effects simply by combining with antigen, or whether direct participation of nonspecific mediators of immunologic damage is required, the antibody molecule was structurally altered by two methods: (1) a globulin fraction of the antiserum was reduced and alkylated to dissociate IgM antibody and destroy the complement-fixing ability of IgG antibody, and (2) a globulin fraction was digested with pepsin to destroy the Fc fragment of IgG. Both modified fractions retained the capacity to produce congenital malformations. This result suggests that complement and other nonspecific mediators are not involved in the immunologic damage which results in malformations in this system. The finding lends support to the proposition that this teratogenic antiserum acts by interfering sterically with normal yolk sac function during a critical period of fetal development.
The Prevention of Autoimmunity to the Thyroid and Allergic Thyroiditis by Antigenic Competition
Authors: McMaster Philip R. B. , Kyriakos Michael
Published: December 2022

When bovine gamma globulin (BGG) is mixed with thyroid extract in Freund's complete adjuvant, the BGG competes with the thyroid extract and inhibits the development of allergic thyroiditis as well as the production of antibody and delayed sensitivity to thyroid extract. The amount of BGG required to inhibit the production of thyroiditis, anti-thyroid antibody and delayed sensitivity to thyroid extract depends in part upon the amount of thyroid antigen used to immunize and in part upon the location of the two antigens.
Conversion of Soluble Immune Complexes into Complement-Fixing Aggregates by IgM-Rheumatoid Factor
Authors: Tesar Joseph T. , Schmid Frank R.
Published: December 2022

Complement fixation was studied after aggregation of soluble immune complexes by IgM-rheumatoid factors (RF). Such aggregates fixed larger amounts of complement than did soluble complexes alone. The extent of this reaction was dependent upon the antigen-antibody ratio in the complex. Complement-fixing ability of those complexes formed near equivalence could be further enhanced by IgM-RF, while those formed in greater antigenic excess with little or no ability to fix complement could be aggregated and converted into complement-fixing units by IgM-RF. The composition of complexes that most readily reacted with RF to form aggregates contained two or more molecules of IgG antibody. Within the aggregate, the major share of complement fixation was mediated by the RF component rather than the IgG antibody. These observations are consistent with the view that RF blocks the binding site on IgG for complement and substitutes its own instead. Under the conditions employed, this substitution resulted in a net increase in complement fixation.
Use of Immunoadsorption Techniques in the Preparation of Chemical Agammaglobulinemia
Authors: Frommel D. , Dupuy J. M. , Litman G. W. , Good R. A.
Published: December 2022

Currently the use of affinity chromatography has been directed toward selective purification of proteins (1). Specific antibodies have been obtained in high yield using columns of Sepharose coupled either to haptens or to protein antigens (2). It is the purpose of this communication to report the application of these newly developed immunoadsorbents to selective removal of immunoglobulins from serum by passage on a Sepharose anti-light chain antibody column.

Preparation of the immunoadsorbent. Antiserum to guinea pig light chain was prepared by immunizing a goat with an IgG light chain pool, emulsified in complete Freund's adjuvant (3). In double diffusion analysis this antiserum reacted strongly with the κ type light chains whereas the precipitin reaction against the λ type of light chains though present was significantly weaker.

Five and six-tenths grams of CNBr (37.5 mg/ml of Sepharose), dissolved in water, were added with stirring at room temperature to 150 ml of packed Sepharose 4 B (Pharmacia Chemicals, Piscataway, N. J.), contained in 300 ml of 0.05 M sodium bicarbonate buffer, pH 9.0.
Ultrastructural Analysis of Hemolysin-Forming Cell Clusters
Authors: Saunders George C. , Hammond William S.
Published: December 2022

Following in vitro stimulation of mouse spleen cells with sheep red blood cell antigens (SRBC) many of the resulting hemolysin-forming cells have been found to be associated with cell clusters (1, 2). We have initiated a series of studies designed to investigate the interrelationships of the various cell types found within these clusters. We report here preliminary morphologic observations of hemolysin-forming cell clusters.

Dissociated mouse spleen cells were cultured in the presence of SRBC using Saunders' (3) modification of Marbrook's (4) technique. After 48 hr of incubation, the spleen cells were carefully recovered and transferred into 15-ml centrifuge tubes which were then placed in an icebucket. Cell clusters were allowed to settle out in the cold for 10 min. The supernatant containing the remainder of the cells and medium was discarded. The recovered cell clusters were analyzed for hemolytic activity by the method of Jerne and Nordin (5).
Lymphocytes and Yellow Fever
Authors: Wheelock E. Frederick , Toy Stephen T. , Stjernholm Rune L.
Published: December 2022

Human lymphoeytes can support the replication of 17-D yellow fever virus when stimulated in vitro with phytohemaggiutinin. Between 7 and 11 days following vaccination with 17-D virus, peripheral lymphocytes lose their ability to support virus replication but regain this capacity by the 25th day post-vaccination.
Electron Microscopic Study of Inhibition of Macrophage Migration in Delayed Hypersensitivity
Authors: Smyth A. Collier , Weiss Leon
Published: December 2022

The migration of peritoneal cells from guinea pigs with delayed hypersensitivity to bacillus Calmette-Guérin was studied by light and electron microscopy. To permit satisfactory morphologic study and monitoring of cell migration, cells were concentrated in clots whose open interstices were accessible to medium. This clot technique, compared to the commonly used capillary tube, provided: 1) better viability and more uniform morphology, 2) better sampling for electron microscopy, 3) more uniform exposure to antigen, and 4) the opportunity to use fewer cells. Suspension cultures of spleen cells were also examined. among peritoneal exudate cells from sensitized animals, purified protein derivative (PPD) induced clumping of macrophages, an increase in the number of their microvilli, and some cell death. Many of the clumped cells bore numerous microvilli which intertwined with those of contiguous cells. No intracellular differences between sensitized and control macrophages or lymphocytes, exposed or not exposed to antigen, were discernible. Sensitized cells exposed to PPD in suspension cultures were disc-shaped and clumped. Control cells were flattened on the flask floor, angulated and usually separated. We believe that both the rounded form of the macrophages and their clumping may contribute to the lack of migration.
Immunosuppression by Leukemia Viruses
Authors: Ceglowski Walter S. , Friedman Herman
Published: December 2022

Spleens of mice infected with Friend leukemia virus (FLV) were assayed for antibody-precursor cells by an indirect cell transfer method. Drug-treated syngeneic mice were used as immunologically inert recipients. Varying numbers of spleen cells from either normal control or FLV-infected animals were injected intravenously into the recipient mice, followed immediately by challenge immunization with sheep erythrocytes. At various times thereafter spleens of the recipients were tested for the number of individual antibody-forming cells, as well as for hemolytic antibody foci, regarded as clusters of progeny antibody-forming cells. There was a significant diminution in the number of both plaque-forming cells (PFC) and foci in recipients of spleen cells from donor mice infected with FLV. The greatest suppression occurred when donors were infected 7 to 14 days before sacrifice. There was a slight to moderate suppression when the donor spleen cells were obtained from animals infected shortly before sacrifice.

The calculated number of antibody-precursor cells present in the donor spleen cell suspension from infected mice was less depressed than would be expected from the suppression observed in the number of PFC appearing directly in the spleens of infected and directly immunized mice. Although the pool of spleen cells capable of responding to antigen and developing into PFC was depleted in spleens of infected animals, it appeared that there was also a secondary effect by virus on the proliferative capacity of such precursor cells and their progeny. The nature and significance of immune suppression at the cellular level are discussed in relation to the mechanism of virus-induced leukemogenesis.
Human Immunologic Response to Soluble Antigens
Authors: Brandriss Michael W. , Bullock Ward E.
Published: December 2022

Volunteers were injected intradermally and subcutaneously with soluble dinitrophenyl-human serum albumin (DNP-HSA) conjugates. Some responded with increasing titers of mercaptoethanol-sensitive agglutinating antibody to DNP-coupled red cells and some developed immediate type wheal and flare skin reactivity. Serum from one of these subjects was capable of passive transfer of DNP-specific wheal and flare reactions.

Thirty-eight of the 55 subjects developed delayed type skin reactions to DNP-HSA. These tended to be maximal in intensity at 24 hr after testing and declined or disappeared by 48 hr in contrast to classic delayed type reactions such as those to tuberculin which are usually prominent at 48 hr. There were also differences in both gross and histologic appearance between DNP-HSA and tuberculin reactions. The response of peripheral leukocytes to antigen in vitro measured by increased DNA synthesis was considerably less to DNP-HSA than it was to tuberculin in individuals who displayed skin reactivity to both. Passive transfer of delayed reactivity to DNP-HSA with dialysates of peripheral leukocytes from sensitive individuals could not be accomplished.

In contrast to the characteristic persistence of delayed sensitivity to tuberculin or other microbial antigens, delayed hypersensitivity to DNP-HSA developed early in the course of sensitization and then declined markedly in intensity or disappeared entirely with time and repeated testing. We suggest that this is a general characteristic of the human immune response to the administration of soluble antigens. This may be a distinct form of immunologic response that can be separated from both the serum antibody response and the classic delayed hypersensitivity that is induced by microbial infections or by administration of antigens in complete Freund's adjuvant.