bluuebunny/crossref_metadata_2025 huggingface dataset:
Antibody Against Neoplastic Plasma Cells
Authors: Watanabe Takeshi , Yagi Yasuo , Pressman David
Published: December 2022

Antigenic differences between mouse myeloma cells and normal mouse lymph node cells were investigated by comparing the reaction pattern of rabbit antisera prepared against cellular components of myelomas of BALB/c mice and of antisera prepared against lymph node cells. The fluorochromasia cytotoxicity test was used. Cells of six individual myeloma lines were used for immunization and cytotoxicity tests. Anti-myeloma sera cytotoxic to myeloma cells of all the lines, yet not cytotoxic to normal lymph node cells, were obtained. The myeloma-cytotoxic antibodies were removed readily by absorption with myeloma sediments, but not at all with sediments of thymus, liver and kidney. Sediments of lymph node and spleen were effective absorbants only when they were used in extremely large amounts. These results indicate the presence of surface antigens on myeloma cells which are not present on lymphocytes, or liver and kidney cells. These antigens are probably present also on normal plasma cells, which comprise a small fraction of the total cell population in the lymph nodes and spleen of normal mice. Although anti-myeloma sera were prepared against individual myeloma lines, all the antisera were cytotoxic to myeloma cells of all the lines. No antigens truly specific to particular lines could be detected even after absorption of antisera with other myelomas. Anti-lymph node sera cytotoxic to lymph node and spleen cells, yet not cytotoxic to myeloma cells, were obtained after appropriate absorption. The lymphocyte-cytotoxic antibodies were removed readily by treatment with lymph node, spleen and thymus sediments. Sediments of myeloma cells and of liver and kidney cells were much less effective, indicating an antigenic difference between lymphocytes and the latter cells.
Destruction of Rabbit Platelets in the Allergic Response of Sensitized Leukocytes
Authors: Siraganian Reuben P. , Osler Abraham G.
Published: December 2022

The ability of sensitized rabbit leukocytes to release histamine from normal platelets is restricted to those cells which alone release histamine on addition of antigen. An excellent correlation between these two leukocyte functions has been obtained. Both reactions have similar metal and antigen requirements as well as parallel kinetics. In both cases cells from hyperimmunized animals are not responsive. Hapten specificity has been demonstrated for both the leukocyte and the leukocyte-dependent platelet response. Cell fractionation studies reinforced the conclusion that only those leukocytes which contain histamine and which release it on the addition of antigen can mediate this leukocyte-dependent platelet response. The lymphocytes, neutrophils and mononuclear cells derived from blood, tissues or exudates contain little or no histamine. Neither can these cells mediate the leukocyteplatelet histamine release. On these grounds it would seem that the sensitized basophil releases histamine on interaction with antigen and, through the mediation of a fluid phase intermediate, causes platelets to release histamine and leak intracellular 86Rb.
Stimulation of Peripheral Lymphocytes by Allogeneic and Autochthonous Mononucleosis Lymphocyte Cell Lines
Authors: Junge Ulrich , Hoekstra John , Deinhardt Friedrich
Published: December 2022

Cultures of peripheral lymphocytes obtained during the acute phase of infectious mononucleosis (IM) were established. Mitomycin C-inhibited cells of these cultures caused abnormally high transformation rates in mixed cultures (MLR) with allogeneic peripheral lymphocytes and to a lesser degree they also stimulated autochthonous peripheral lymphocytes obtained from the same persons 3 to 9 months after their acute disease. This stimulation seems not to be caused by Epstein-Barr virus (EBV), present in the IM-cultures, because concentrated EBV preparations did not stimulate the convalescent lymphocytes and allogeneic lymphocytes from infants with and without circulating EBV antibodies responded equally well in the MLR with cultured IM lymphocytes. Circulating antibodies against surface antigens of autochthonous acute phase IM lymphocytes, stored in liquid nitrogen or cultured, were demonstrated in convalescent IM sera by the indirect membrane fluorescence technique. Blastogenic activity for both allogeneic and autochthonous lymphocytes was found in the cell-free supernatants of these IM lymphocyte cultures and was probably associated with cell membrane fragments.
Use of Chromium Chloride As the Coupling Agent in a Modified Plaque Assay Cells Producing Anti-Protein Antibody
Authors: Sweet George H. , Welborn Frederic L.
Published: December 2022

A method is described for the detection and enumeration of mouse spleen cells producing antibody to protein antigens. The method employs chromium chloride as the coupling agent in a modified plaque technique and offers the advantages of greater economy, ease, speed and reproducibility over other available procedures.
Antigenic Competition Between Heterologous Erythrocytes
Authors: Gershon Richard K. , Kondo Kazunari
Published: December 2022

Antigenic competition between sheep red blood cells (SRBC) and horse red blood cells (HRBC) was shown to be thymus-dependent. Mice deprived of thymus-derived lymphocytes (T cells) were not affected in their response to HRBC by a prior injection of SRBC, even after adjuvant treatment. Mice reconstituted with 6 × 107 thymocytes and inoculated only with HRBC made more anti-HRBC antibody than did mice reconstituted with 1.5 × 107 thymocytes. However, after a prior injection of SRBC the situation was reversed: mice with more T cells now made less anti-HRBC antibody than did mice with fewer of these cells. The addition of various lymphoid cells to mice undergoing antigenic competition could not overcome it; indeed, the addition of thymocytes magnified it. Normal spleen cells given to mice lethally irradiated 2 days (but not 6 days or 1 hr) after the inception of an anti-SRBC response were impaired in their ability to respond to HRBC. The amount of interference produced by SRBC on the subsequent response to HRBC did not correlate with the amount of anti-SRBC antibody made. We suggest that the most likely interpretation of these data is that T cells make, in response to antigenic stimulation, a factor which temporarily inactivates other immunocompetent cells.
Differential Effects on Immunogenicity and Antigenicity of Heat, Freezing and Alkali Treatment of Bacterial Antigens
Authors: Whang H. Y. , Mayer H. , Neter E.
Published: December 2022

Heating, freezing and treatment with alkali of common enterobacterial antigen and of endotoxic lipopolysaccharides obtained by the aqueous ether procedure strikingly affected their immunogenicity (antibody response of the rabbit), but not their antigenicity (antibody-neutralizing capacity). Immunogenicity of these antigens was reduced by more than 90% by heating for 1 hr at 100°C or treatment with 0.25 N NaOH. Immunogenicity was restored to a significant degree after repeated freezing (-20°C) and thawing of heated antigen and to a lesser degree after alkali treatment. Antibody-neutralizing capacity remained unaffected by these measures. Lipopolysaccharides extracted by the phenol-water procedure had poor immunogenicity, which was affected only slightly by heating or freezing. Immunogenic lipopolysaccharides consisted of two fractions separated by Millipore filtration (0.22 µ); the aggregated fraction engendered antibodies readily and the soluble fraction did not. Heated, alkali-treated and filtered preparations of these antigens induced immunologic priming even when antibody production, as measured by the hemagglutination method, was minimal.
“Nonspecific” Stimulation of Lymphocyte Transformation by Cellular Fractions and Acid Extracts of Group a Streptococci
Authors: Keiser Harold , Kushner Irving , Kaplan Melvin H.
Published: December 2022

Hot hydrochloric acid extracts of group A streptococcal cells of types 5, 12 and 19 induced “nonspecific” blast-transformation and increased thymidine-14C incorporation by adult and cord blood lymphocytes of normal human donors. A similar degree of transformation was induced by preparations of streptococcal cell walls and cell membranes and by hot acid extracts of these cell components. Streptococcal cytoplasm was found to contain an inhibitor of phytohemagglutinin-induced transformation and of spontaneous transformation of cord blood lymphocytes. Treatment of streptococcal cytoplasm with hot hydrochloric acid resulted in the appearance or release of lymphocyte transforming activity in such preparations. The lymphocyte transforming activity of streptococcal acid extracts was unaffected by digestion with ribonuclease, trypsin, chymotrypsin, and pepsin. After gel filtration on Sephadex G-75, acid extracts of streptococcal cell membranes, cell walls and cytoplasm all exhibited their major transforming activity in a peak with maximum at Kd 0.36, suggesting a similar average molecular size for the transforming agent or agents derived from these components. The “nonspecific” mitogenic activity of acid extracts of streptococcal cells resembled that of streptolysin S preparations with respect to enzyme susceptibility and dose-response. Hot acid-treated streptolysin S preparations retained most of their transforming activity, while their hemolytic activity was completely destroyed. These data raise the possibility that the transforming activities of streptococcal cell acid extracts and of streptolysin S preparations are attributable to a similar or identical constituent.
Proliferation of Maximally Activated Antigen-Primed Cells: A Computer Simulation
Authors: Cohen Stanley , Milgrom Martin
Published: December 2022

Most reported models of formation of antibody or induction of tolerance require interaction between antigen and an antibody-like receptor on the surface of a potentially immunocompetent cell as a critical step for induction or suppression of the immune response. In the simplest situation, where interaction is between identical antigenic fragments and identical receptor units, the nature of the immune response must depend upon the geometric distribution of bound receptors on the cell. Considerations such as these led to formulation of a precise, quantitative model of the process by which the immune response is activated. The availability of such a model made possible computer simulation of the division of an activated cell to form a clone. In this simulation, if activated cells were allowed to divide until the number of antigen-bound receptor sites fell below various threshold values, the frequency distribution of progeny as a function of bound sites was biphasic for certain narrow ranges of these thresholds. The two populations so defined, which differed in the number of divisions they had undergone and the number of bound receptors they possessed, might represent a pool of end-stage antibody-producing cells and a pool of intermediate cells that have immunologic memory. A major consequence of this interpretation was restriction of heterogeneity in the secondary response.
The Effect of Transplanted Lymphoid Cells, Irradiation, and Drug Therapy on Permanent Immunologic Tolerance
Authors: Larsen Jørgen Hannover
Published: December 2022

Abolition of the permanent and constant tolerance to lymphocytic choriomeningitis (LCM) virus in inbred mice has hitherto only been possible by transplantation of syngeneic, sensitized or unsensitized lymphoid cells to the tolerant virus-carriers. A series of experiments showed that no essential difference exists between lymphoid cells isolated from the spleens, lymph nodes or buffy coat of hyperimmune, syngeneic mice in their ability to abolish the tolerance of virus-carrier recipients after transplantation. Large numbers of unsensitized thymus cells were without effect. Sensitized bone marrow cells only had a slight effect. Fetal liver cells were without effect. Using the same system with transplantation of lymphoid cells an Eichwald-Silmser effect was found in the highly inbred strain of C3H mice that was used. Large doses of transplanted, killed, syngeneic or living xenogeneic cells had no effect on the tolerance, nor did x-irradiation or treatment with cortisone or methotrexate. Syngeneic, sensitized lymphoid cells in Millipore chambers implanted into tolerant virus-carriers conferred the ability to produce antibody, but the viremia was not essentially affected.
Murine Malaria: The Role of Humoral Factors and Macrophages in Destruction of Parasitized Erythrocytes
Authors: Criswell B. Sue , Butler William T. , Rossen Roger D. , Knight Vernon
Published: December 2022

Interactions between cellular and humoral factors in the response of mice to malaria were evaluated by placing Millipore diffusion chambers (0.22 µ pore size) containing washed macrophages from mice and mouse erythrocytes parasitized with Plasmodium berghei in the peritoneal cavities of BALB/c- male mice that were normal or had acute or chronic malaria. Macrophages were obtained from normal mice, endotoxin-treated mice, quinacrine-treated mice, or mice with chronic malaria established by treatment with quinacrine hydrochloride. Parasitized erythrocytes were obtained from mice with acute malaria. The rate of destruction of parasitized erythrocytes alone in Millipore chambers was greater when chambers were placed in mice with chronic malaria than when placed in normal mice. This finding suggested that humoral factors diffused into the chamber causing increased destruction of parasitized erythrocytes. When macrophages from mice with chronic malaria were added to the chambers with parasitized erythrocytes, and implanted into normal mice, an enhanced rate of clearance of parasitized erythrocytes also occurred. Macrophages from normal mice or from endotoxin-treated mice did not increase the rate of clearance of parasitized erythrocytes when placed in normal mice. However, an accelerated rate of clearance of parasitized erythrocytes occurred when chambers containing parasitized erythrocytes and macrophages from normal mice or mice with chronic malaria were placed into mice with chronic malaria. Therefore, humoral factors not only acted directly on the chamber contents to cause destruction of parasites, but also possibly acted synergistically with macrophages to enhance destruction of parasitized erythrocytes.
Specificity of Anti-Poly G-Poly C Antibodies
Authors: Nahon-Merlin E. , Michelson A. M. , Verger C. , Lacour F.
Published: December 2022

Immunization of rabbits with the double-stranded polynucleotide complex poly G·poly C adsorbed to methylated bovine serum albumin produced antibodies that reacted with RNA. These antibodies were specific for ribosomal RNA from animal cells and RNA from certain viruses.
Immunologic Enhancement in Malaria Infection (Plasmodium Berghei)
Authors: Jerusalem Christoph , Weiss Margaret L. , Poels Lambert
Published: December 2022

Immunologic enhancement of malaria (Plasmodium berghei) comparable to that described in tumor and transplantation immunity could be produced by a variety of active and passive immunization procedures, using quantities of antigen comparable to those commonly employed. Much greater quantities were required for the development of some degree of protection. The clinical importance and the possible relationship to tumor and transplantation immunity, and the possible mechanisms of this phenomenon, are discussed. Attempts at active and passive immunization against malarial parasites not only frequently failed to show beneficial results, but often led to exacerbation of the challenge infection. The challenge infection is influenced by various antibodies in different proportions. Non-cytotoxic antibody appears to act as an enhancing agent in the absence of a second, protective antibody; but it may play a minor role, or even be a decisive factor, in the presence of a protective antibody. The observed enhancement phenomenon appears to resemble the immunologic enhancement of tumor growth.
PCA Reactions in Adult Specific Pathogen-Free Chickens
Authors: Ettinger A. Conway , Hirata A. A.
Published: December 2022

Young chickens readily gave PCA reactions but as they became older they gradually lost this capacity (4). PCA reactions were elicited in SPF chickens ranging from 45 to 897 days of age. When these SPF chickens were later housed in an ordinary environment for more than 6 months, they retained their ability to give PCA reactions.
The Unique Molecular Weight of the Heavy Chain from Human IgG3
Authors: Saluk Paul H. , Clem L. William
Published: December 2022

Human IgG consists of four subclasses (IgG1, IgG2, IgG3 and IgG4) which are known to differ from each other in certain antigenic, structural and biologic properties (1–5). In particular, the IgG3 subclass has many unique properties and it is catabolized faster than the other three subclasses (6, 7). Most monoclonal IgG cryoglobulins with anti-IgG activity belong to this subclass (8) as do IgG myeloma proteins that exhibit marked concentration- and temperature-dependent aggregation (9). In addition, IgG3 is the only subclass which does not react with staphyloccal protein A (10). One interesting structural feature unique to IgG3 is five inter-heavy chain disulfide bonds (11).

Recently we found that gel filtration of a IgG3 cryoglobulin on a Sephadex (Pharmacia Fine Chemicals, Piscataway, N. J.) G-200 column at pH 3.9 yielded an elution position ahead of human Cohn fraction II (FII).
Ultrastructural Study of Clq.
Authors: Shelton E. , Yonemasu K. , Stroud R. M.
Published: December 2022

Purified C1q was diluted in buffer to between 10 and 50 µg/ml and droplets were placed on carbon-coated 400-mesh copper grids. Some grids were rinsed with dilute buffered formaldehyde or glutaraldehyde in order to fix the molecules. All grids were rinsed and stained in either 2% potassium phosphotungstate or 1% uranyl acetate. Approximately 200 photomicrographs from six different preparations were analyzed.

The whole C1q molecule is made up of an undetermined number of dense subunits which measure, on the average, 65 to 70 Å in diameter. These sub-units are joined at a central attachment area by strands estimated to be 60 to 100 Å in length and 15 to 20 Å in width. The structure of the central attachment area is hard to define, but is definitely less complicated than the central region of the IgM molecule.
Cleavage of Human Cls̄ by Proteolytic Enzymes: Evidence for a C4-Binding Site of Cls̄ Distinct from the Catalytic Center
Authors: Taubman S. B. , Lepow I. H.
Published: December 2022

Previous reports have shown that trypsin, plasmin and lysosomal enzymes from human peripheral leukocytes generate esteratic activity from human C1s. With continued incubation, esteratic activity progressively diminishes. Present studies demonstrated that C1s̄ was cleaved into at least two fragments during incubation with these proteolytic enzymes. Only the heavier fragment retained esteratic activity; the lighter fragment increased in concentration with incubation time. Inactivation kinetics of C1s̄ were first-order both with respect to esteratic activity and residual ability to inactivate purified human C4 in solution. However, the rate of the latter reaction was much greater than the former and the rapid loss of ability to inactivate C4 was associated with C1s̄ cleavage, suggesting the presence of a C4-binding site on native but not cleaved C1s̄. Indeed, a complex of C4 and native, but not cleaved, 125I-C1s̄ could be demonstrated by sucrose density gradient ultracentrifugation.
Characterization of the C3 Inactivator of Human Serum
Authors: Cooper N. R.
Published: December 2022

This study was initiated to investigate the properties and mechanism of action of the C3 inactivator, a serum protein which inhibits the hemolytic activity of cell bound C3. The C3 inactivator was found to be inseparable from the C3 proactivator on purification, and the final preparations, homogeneous on disc- and on immunoelectrophoresis, possessed both activities.

Five separate preparations were isolated which differed slightly in charge and markedly in functional properties, suggesting the existence of several distinct functional sites in the proactivator molecule. These preparations are: 1) 5–6S β-pseudoglobulins; 2) antigenically indistinguishable on Ouchterlony analysis with specific antisera; 3) able to hydrolyze certain synthetic amino acid esters, such as acetyl glycyl-l-lysine methyl ester; 4) fully able to complex with cobra factor. These preparations, however, differ markedly in: 1) ability to cleave C3 in free solution in the presence or absence of cobra factor, 2) activity/protein ratios in the C3 inactivator assay and 3) ability to inhibit the hemolytic activity of C3 proactivator depleted serum.
Generation of Chemotactic Activity from Human Serum and Purified Components of Complement by Serratia Proteinase
Authors: Chapitis J. , Ward P. A. , Lepow I. H.
Published: December 2022

The ability of culture filtrates of Serratia marcescens to effect immunoelectrophoretic (IEP) conversion of C3 in human serum in the presence of EDTA or of purified C3 correlated with elaboration of Serratia proteinase. This was confirmed with purified proteinase (supplied by Dr. C. Decedue). Higher concentrations of purified proteinase were required for partial IEP conversion of C3 in serum (500 µg proteinase/ml) than purified C3 (1 to 2 µg/ml), indicating the presence of a serum inhibiter which was localized in a 36% to 57% saturated ammonium sulfate fraction of pseudoglobulin. The extent of IEP conversion of purified C3 was a function of proteinase concentration and incubation time and was associated with loss of hemolytic activity and cleavage of C3 with appearance of two light fragments (estimated molecular weights 34,000 and 5,600). Chemotaxis for rabbit neutrophils was found in the slowly sedimenting region of sucrose gradients.
Activated C5: Its Stabilization by Complexing with C6 and the Catalytic Role of Cell-Bound C2a in Its Formation
Authors: Goldlust M. B. , Shin H. S. , Mayer M. M.
Published: December 2022

In the course of our work on the formation and properties of the EAC1̄,,5b,6 intermediate, we have found that eluates prepared from these cells contain a complex factor which has an apparent sedimentation rate of 14S to 15S and comprises C5b and C6. This material, designated C5b/6, converts EAC ∼ 3 (EAC or EAC4,3b) to EAC ∼ 6 (i.e., an intermediate susceptible to lysis by C7, C8 and C9). The conversion proceeds in the presence of EDTA. In contrast, native C5 and C6 will react only with EAC to yield EAC ∼ 6, i.e., cell-bound C2a is essential for the reaction with native C5 and C6. The lack of C2a requirement in the reaction with C5b/6 is supported by the observation that monospecific rabbit anti-guinea pig C2 does not inhibit C5b/6 actionon EAC ∼ 3. On the other hand, as would be expected, monospecific rabbit anti-guinea pig C5 effectively blocks conversion of EAC ∼ 3 to EAC ∼ 6 by C5b/6.
Detection of Antibodies to the Basic Protein of Human Myelin by Radioimmunoassay and Immunofluorescence
Authors: Lennon Vanda A. , Whittingham Senga , Carnegie Patrick R. , McPherson Thomas A. , Mackay Ian R.
Published: December 2022

Specific antibodies to a basic protein that is a component of myelin were demonstrated quantitatively by gel filtration radioimmunoassay in three species, rabbit, guinea pig and rat, after a single immunization with the antigen in complete Freund's adjuvant. In the rabbit, the antibody to 125I-labeled basic protein was IgM at the peak primary response and IgG late in the response after a second immunization. There was good correlation between the results of two methods for detecting this antibody, namely, radioimmunoassay and indirect immunofluorescence using unfixed frozen sections of rabbit central nervous tissues. Fluorescence with anti-basic protein serum was specific for the basic protein and occurred predominantly in areas containing myelinated fibers. The speckled pattern contrasted with the more diffuse pattern given by an anti-myelin serum, suggesting restricted exposure of the antigenic determinants of the basic protein in the intact central nervous system. Following immunization, more than 85% of guinea pigs had measurable levels of antibody that were correlated with time after immunization, but not with immunizing quantity of basic protein. In the Lewis rat, the serum antibody level and severity of clinical signs of experimental autoimmune encephalomyelitis (EAE) were proportional to the quantity of basic protein. The lowest quantity induced clinical and histologic EAE but no measurable antibody. There was no clear correlation between presence or levels of antibody to basic protein and neurologic disease in any of the species studied. The determinant(s) of the basic protein molecule that induce antibody production, as yet not identified, were apparently not in the same region of the molecule as the main encephalitogenic determinant.
Studies on Antigen-Antibody Complexes
Authors: Arend William P. , Mannik Mart
Published: December 2022

The quantitative tissue uptake of soluble immune complexes made with rabbit γG antibodies, was studied in normal and complement-depleted rabbits. The immune complexes that were cleared quickly from the circulation were primarily localized in the liver; less than 1% of the injected complexes was found within the lungs, kidneys or spleen. The tissue distribution of complexes made with intact antibodies was unchanged in complement-depleted rabbits. Soluble complexes made with reduced and alkylated antibodies fixed complement ineffectively. There was no hepatic uptake of HSA-anti-HSA and decreased uptake of γG-anti-γG and γM-anti-γM complexes, made with reduced and alkylated antibodies. In the latter two instances residual hepatic uptake was due to human γG or γM antigens in the complexes. It was concluded that the in vivo interaction of the fixed tissue macrophages and γG class of antibodies in soluble immune complexes was not mediated by circulating complement components. However, studies with a Waldenström's macroglobulin suggested that the in vivo interaction of macrophages and γM depended upon the third component of complement. It appeared that the complexes had to contain more than two γG antibody molecules for rapid uptake by the RES to occur. The RES uptake of complexes did not depend on their physical size. The rapidly removed complexes fixed complement strongly in vitro, although their in vivo uptake by the RES did not depend on an intact complement system in the circulation.
Conversion of Non-Immune Rabbit Spleen Cells by Ribonucleic Acid of Lymphoid Cells from an Immunized Rabbit to Produce IgM and IgG Antibody of Foreign Heavy-Chain Allotype
Authors: Bell Clara , Dray Sheldon
Published: December 2022

Spleen cells (SpC) from non-immunized rabbits homozygous for one allotype at the a heavy-chain locus of immunoglobulins were incubated with RNA extracts of lymph nodes from immunized homozygous rabbits of a different genotype to yield IgM “direct” plaque-forming cells (PFC) or IgG “indirect” PFC. The heavy-chain allotype of the antibody in the plaques was identified by radioautography and by inhibition of plaque formation using anti-allotype antibodies. The allotype of the RNA donor rabbit was identified in 75% to 86% of the IgM “direct” plaques and in 43% to 62% of the IgG “indirect” plaques. The allotype of the SpC donor rabbit was found in 2% to 12% of the IgM “direct” plaques and in 24% to 39% of the IgG “indirect” plaques. The lysates of “RNA-converted” SpC were analyzed for immunoglobulins of each heavy-chain allotype by immunodiffusion. Allotypes of both RNA and SpC donor rabbits were found in the lysates, whereas only that of the SpC donor was found in the lysates of untreated SpC, and neither allotype was found in the RNA extracts. These results were similar to those reported previously for light-chain allotypes and indicated that a component of the RNA extract provided information for synthesis of at least part of the heavy chains as well as the light chains of the IgM and IgG antibody molecules.
Genetic Control of Immune Responsiveness to Limiting Doses of Proteins and Hapten Protein Conjugates in Guinea Pigs
Authors: Green Ira , Benacerraf Baruj
Published: December 2022

As shown earlier for responsiveness to limiting doses of bovine serum albumin (BSA), strain 2 and not strain 13 guinea pigs form antibody when immunized with low doses of human serum albumin (HSA), an antigen strongly cross-reactive with BSA. (2 × 13)F1 animals responded as did the parental strain 2 animals, indicating dominant genetic control of responsiveness to limiting doses of BSA and HSA. Responsiveness to low doses of BSA in (2 × 13)F1 × 13 back-cross offspring was associated with inheritance of the PLL gene and also of the major strain 2 histocompatibility specificities. Genetic control of the immune response to BSA and HSA in strain 2 and strain 13 animals was specific, as no significant difference was observed in the antibody response of these guinea pigs to limiting doses of several other unrelated proteins. Random-bred Hartley guinea pigs immunized with low doses of HSA were distributed into distinct populations with high or low anti-HSA antibody responses. Responsiveness to limiting doses of HSA in these random-bred animals was not associated with possession of the PLL gene. Antihapten antibody response depended upon the gene controlling responsiveness to the carrier molecule in strain 2 and strain 13 guinea pigs immunized with limiting doses of DNP7-BSA. Strain 2 guinea pigs formed considerably higher levels of anti-DNP antibody than strain 13 guinea pigs in response to this antigen. In contrast, strain 13 guinea pigs developed significantly higher anti-DNP serum antibody levels than strain 2 animals when immunized with DNP6-GPA.
Streptococcal Related Glomerulonephritis
Authors: Markowitz A. S. , Horn Donald , Aseron C. , Novak Robert , Battifora H. A.
Published: December 2022

Experimental glomerulonephritis was induced in rhesus monkeys both actively and passively. The actively induced disease was brought about by immunizing the animals with alum-precipitated PGT CM (soluble glycoprotein from the nephritogenic streptococcal protoplasmic membrane) and the passively induced disease by intravenous injection of rabbit anti-PGT CM γ globulin-rich fraction. In both instances the disease was moderate in intensity and relatively short in duration. Of special interest however, was that in both instances the γ globulin (whether heterologous or autologous) and C3 deposition was grossly granular and located in the mesangium. Analytical and preparative acrylamide gel electrophoresis of PGT CM would tend to explain the moderate disease induced in that only a small portion of the immunogen employed (12% or less) is cross-reactive with glomerular basement membrane.
Association of Polypeptides with Ribonucleic Acid
Authors: Egan Marianne L. , Smyth Robert D. , Maurer Paul H.
Published: December 2022

Peritoneal exudate (PE) cells were incubated with synthetic polypeptides and the association of antigen with extracted cellular RNA was determined. Positively charged polypeptides were in higher concentration in the RNA fraction than were negatively charged polymers. Neutral polymers were associated to the least extent (0 ng to 0.2 ng/µg RNA). Incubation of insoluble methylated bovine serum albumin (MBSA) complexes of a negatively charged polymer (GAT) with PE cells resulted in RNA-associated polypeptide concentrations equivalent to those observed with the positively charged polymer (LAT) without a carrier protein. The levels of RNA-associated antigen were related to concentration of polypeptide in the incubation mixture. GAT was associated with RNA or PE cells from GAT-tolerant rabbits to a greater extent than with RNA from cells of normal or GAT-immune animals. Lymph node, spleen, bone marrow or testes cells exhibited no markedly different degree of pattern of association with the various RNA fractions. Association of antigen with low molecular weight RNA was demonstrated by methylated albumin-kieselguhr column chromatography and sucrose gradient ultracentrifugation.
Inhibition of Histamine and SRS-A from Monkey Lung Tissue by Chlorophenesin
Authors: Malley A. , Baecher L.
Published: December 2022

Chlorophenesin completely inhibited allergen-reagin-induced histamine and SRS-A release from monkey lung tissue. Preliminary studies suggest that chlorophenesin mediates this inhibition by activating adenylcyclase.
Combined Effect of Phytohemagglutinin and Antithymocyte Serum on Skin Xenograft Survival in Mice
Authors: Chen C. M. C. , Chung C. F. , Tsai L. C.
Published: December 2022

Phytohemagglutinin (PHA) is generally regarded as a weak immunosuppressive agent. Calne et al. (1) initially reported that it can potentiate the immunosuppressive action of azathioprine in dogs with renal allografts when the two substances were given together. Later on, Gertner et al. (2) recorded again that combination of PHA with either azathioprine or prednisone increased markedly the survival time of skin allografts in dogs and decreased the need for toxic doses of azathioprine and prednisone. Recently, Stefani and Moore (3) suggested that there might be a synergistic effect of PHA and antilymphocyte serum (ALS) in preventing allograft rejection in the clinical situation. In view of the previous evidence that PHA can significantly prolong the survival time of skin xenografts in mice (4) and that antithymocyte serum (ATS) has a greater immunosuppressive effect than ALS on allograft survival (5, 6), it seemed justifiable to do a preliminary experiment concerning skin xenografts in mice treated with a combination of PHA and ATS to see whether a significant synergistic effect could be obtained in this model.
Method of Serum IgA Isolation
Authors: Mestecky J. , Kulhavy R. , Kraus F. W.
Published: December 2022

Although immunoglobulin A (IgA) is the second most abundant immunoglobulin in human serum, the isolation of large quantities of it in immunochemically pure form has been difficult. Fractionation procedures for purifying serum IgA have recently been reviewed by Vaerman (1). Because of the similarity between IgA and IgG, techniques based on net-charge and molecular-weight differences between the two give a low yield. Other methods, based on specific antigenic determinants of IgA, require a monospecific antiserum and its attachment to form an immunoadsorbent.

The method reported here used the F(ab′)2 fragment of antibody to IgA; the immune precipitate was dissociated in low pH buffer and the components were subsequently separated by gel filtration.

An antiserum specific for IgA was produced in the goat. Immunochemically pure secretory IgA (S-IgA) from human colostrum was mixed with complete Freund's adjuvant (2) and injected both subcutaneously and intramuscularly. Three immunizations, of 20 mg S-IgA each, were given 3 weeks apart.
Production and Evaluation of a Formalin-Killed Chikungunya Vaccine
Authors: Harrison V. R. , Eckels K. H. , Bartelloni P. J. , Hampton C.
Published: December 2022

Production methods and immunogenic characteristics of a formalin-inactivated Chikungunya vaccine prepared in bank-frozen green monkey kidney tissue culture were described. The total absence of untoward reactions or side effects and the excellent immunogenic response in volunteers attested to the safety and immunogenicity of this vaccine.
Studies of the Mechanism of Binding of Chemically Modified Cytophilic Antibodies to Macrophages
Authors: Thrasher Susan G. , Cohen Stanley
Published: December 2022

Chemical modification of lysine or tryptophan groups of rabbit γG antibody led to marked reduction in its cytophilic binding ability. The procedures used, carbamylation, amidination and benzylation, were highly selective in their effect on the various properties of antibody. Thus, although they led to loss of complement-fixing ability and diminution in the rate of immune aggregation, they had no significant effect on the primary union of antibody with antigen or on the ability of antibody to mediate anaphylaxis in the guinea pig. This selectivity suggested that the modifications led to specific alteration of Fc binding sites rather than nonspecific alteration of the molecule as a whole. The fact that these chemical alterations affect both cytophilic binding and complement fixation does not imply a functional relationship between the two. Cytophilic binding did occur in the absence of complement. Moreover, the various treated antibodies had dissociation of these activities; concentrations of conjugated antibody sufficient for cytophilic rosette formation were not capable of fixing enough complement for detectable erythrocyte lysis. In this regard, differences in the ability of untreated antibody to mediate rosette formation and complement fixation in the direct and indirect procedures for cytophilic binding suggest that the mechanisms of complement-antibody and macrophage-antibody interaction are distinct, even though both reactions can be a property of the same antibody molecule.
Genetic Control of Immune Responsiveness to a Glutamic Acid, Alanine, Tyrosine Copolymer in Mice
Authors: Martin W. J. , Maurer P. H. , Benacerraf B.
Published: December 2022

Mice of various strains were immunized with a random terpolymer of glutamic acid60 alanine30 tyrosine10 (GAT10). The ability to produce significant anti-GAT10 antibody, detected by an antigen-binding assay, was possessed by mice of H-2a, H-2b, H-2d, H-2k histocompatibility alleles. Mice with H-2p and H-2s alleles were non-responders. Congenic mice that differed only at the H-2 locus responded according to their H-2 type.
IgE Levels in Normal Infants and Mothers and an Inheritance Hypothesis
Authors: Bazaral Michael , Orgel H. Alice , Hamburger Robert N.
Published: December 2022

Immunoglobulin E (IgE) in serum was measured by an improved quantitative test (an assay with competitive inhibition of the binding of 125I PS-IgE to bromacetylcellulose-anti-ND-IgE).

IgE levels were determined in serum samples from 35 postpartum mothers (μ = 205 units/ml, range 19.4 to 810), 33 newborn cord sera (μ = 2.1 units/ml, range <1 to 7.5), 23 6-week-old infants (μ = 5.5 units/ml, range <1 to 31.6) and 17 6-month-old infants (μ = 57.6 units/ml, range 4.1 to 458). In 28 maternal-infant pairs we confirmed the absence of transplacental passage of IgE. Cord serum levels reported here are lower than those from other laboratories; in our study half of the newborns had less than 1.5 units/ml of IgE. No synthesis of IgE was apparent in the serum of one-third (8) of the 6-week-old infants, and only 3 of 23 overlapped the adult range. All 6-month-old infants had measurable IgE levels. The 3 with clinical allergy had 3 of the highest 4 serum IgE levels (47, 69, and 458 units/ml). In the adult group there was a presumptive fit to the Hardy-Weinberg distribution, consistent with simple Mendelian heredity of basal IgE levels.
Immunoprecipitin Reaction of Influenza Virus-Antibody Complex with Anti-IgG
Authors: Daugharty Harry
Published: December 2022

A virus immunoprecipitin radioassay (VIR) was developed to quantitate influenza virus interaction with specific antibody. After the initial virus-antibody reaction, quantitative binding of isotopically labeled anti-IgG resulted in the final formation of a virus immunoprecipitate. The anti-IgG radioactivity incorporated in this virus immunoprecipitate was a measure of the variation in any one of the reactants. This finding confirmed that the immunologic specificity of the VIR procedure was directly dependent upon virus antigen concentration and virus antibody dilution for proportional binding of the 125I-labeled anti-IgG with no prozone activity. Competitive inhibition of labeled anti-IgG antibody by unlabeled anti-IgG for detecting specifically bound virus antibody indicates the sensitivity of the virus immunoprecipitin reaction. Alternate methods for eliminating unbound anti-IgG from the virus immunoprecipitin complex, the carrier precipitate and the sucrose gradient methods were used. In the sucrose gradient method, the changes in the reactants, as evidenced by sedimentation behavior, were quantitative and proportional to the dilution of influenza antibody. Hemagglutination inhibition is no more reproducible and is not as sensitive or precise as the VIR for quantitating influenza-specific antibody. Correlation between the two tests indicates that they probably measure the same factor of influenza-specific antibody.
Studies on the Immune Response to Penicillin and Cephalothin in Humans
Authors: Spath Peter , Garratty George , Petz Lawrence
Published: December 2022

This study evaluated the critical factors in the titration of penicillin and cephalosporin antibodies using hemagglutination techniques. Emphasis was placed on the development of sensitive and reproducible methods for further investigational work as well as the development of a simple and practical method suitable for use by clinical laboratories with only occasional need for performing such tests. Conditions for optimal sensitization of erythrocytes with the antibiotics were evaluated including concentration of the antibiotics; pH, temperature, and time of sensitization; comparison of red cells from different donors; and the effects of varying conditions of storage and age of the red cells. Optimal sensitization of red cells by penicillin are obtained when 1 ml of fresh erythrocytes is incubated with 1,000,000 units Kbenzylpenicillin G in 15 ml Trimethylamine (TMA)-buffer, pH 10.0, at room temperature for 2 hr. Similarly, for preparation of cephalothin-sensitized red cells, 1 ml of fresh erythrocytes is incubated with 400 mg Na-cephalothin in 10 ml TMA-buffered saline (pH 10.0) at 37°C for 2 hr. Red cells sensitized at pH 10.0 with penicillin gave titers 30 times higher than those sensitized at pH 7.3, but only marginal differences were noted with cephalothin. Several hemagglutination systems were evaluated, including standard blood-banking methods such as saline, albumin and antiglobulin techniques. These were compared with agglutination in a diluent containing dextran, normal rabbit serum, and Tris-buffered saline (Dex-NRS-TBS). The indirect antiglobulin (Coombs') test using antibiotic-sensitized cells is optimal for clinical purposes. However, if numerous penicillin and cephalothin antibody titrations are to be performed for investigational work, direct agglutination in Dex-NRS-TBS is generally more practical.
Antigenicity of Alum and Aqueous Adenovirus Hexon Antigen Vaccines in Man
Authors: Kasel J. A. , Couch R. B. , Douglas R. G.
Published: December 2022

The availability of similarly prepared hexon antigen vaccines of three different serotypes of adenovirus (types 1, 2 and 5) provided the opportunity to systematically examine the responses in man to several variables of subunit vaccine and vaccination. Each of the vaccines exhibited differing antigenicity for guinea pigs and was available as an aqueous and an adjuvant preparation. In addition to examining the effect of these variables of the vaccine, the effect of one and two injections and route of inoculation were also examined.

Recent studies with diphtheria toxoid in aqueous and adjuvant form have suggested that the physical state of antigen is of paramount importance for development of 11S IgA secretory antibody (1). In those studies, the aqueous diphtheria toxoid preparation stimulated secretory antibody following topical as well as parenteral administration. However, when adjuvant vaccine was given parenterally, volunteers developed high serum antibody titers but no 11S IgA secretory antibody and when given topically this vaccine was apparently non-antigenic.
Variable Region Sequence Differences Associated with Rabbit Light Chain Allotypes
Authors: Hood L. , Waterfield M. , Morris J. , Todd C. W. , Eichmann K. , Krause R.
Published: December 2022

The light chains from eight individual rabbits, two from each of the four allotypes, were run on the automatic protein sequencer for 21 residues. The resulting PTH-amino acid derivatives were quantitated by gas chromatography and after hydrolysis with the amino acid analyzer. It is possible to distinguish unequivocally the light chains from each of the four allotypes. These differences include striking shifts in the ratios of VK subgroups expressed (e.g., the b6 light chains seem to be predominantly of the VKI subgroup whereas b4's and b5's express primarily the VKII subgroup). These differences include amino acid substitutions (e.g., b9 chains show predominantly a serine at position 9 whereas the other subgroups express predominantly an alanine at this position). Thus in rabbit light as well as heavy chains, V region sequences correlate with allotype differences. Obviously light chains offer a much simpler system than heavy chains for studying the structural basis for V region sequence differences and rabbit allotypy.
Possible Role of Antibody Specificity in Genesis and Survival of Plasmacytomas
Authors: Leon Myron
Published: December 2022

The specificity of a myeloma protein may play a role in the genesis and/or the survival of a plasmacytoma. If the immunoglobulin synthesized by a plasma cell cross-reacts with an appropriate determinant present inside the cell it may cause alteration of cellular metabolism. Such an altered cell may be more prone to neoplastic transformation than a normal plasma cell. If secreted myeloma protein cross-reacts weakly with external membrane determinants, it may act as an enhancing antibody, protecting the plasmacytoma from the normal cytotoxic surveillance system. Plasmacytomas secreting antibodies with such specificities may thus have a higher rate of survival than plasmacytomas secreting immunoglobulins that have no affinity for plasma cell membranes.
Hemolytic Anemia in Agammaglobulinemic Chickens: A Model of “Autoimmune” Disease in Immune Deficiency
Authors: Weiden Paul L. , Blaese R. Michael
Published: December 2022

Agammaglobulinemic chickens, which have less than 0.5% of normal immunoglobulin levels, develop a hemolytic anemia with variable frequency. Autologous red blood cell (RBC) survival studies using DFP-3H-labeled cells demonstrated a survival of 15 days in agammaglobulinemic anemic chickens compared with 30 days in normal chickens. Homologous survival studies demonstrated normal survival of normal RBC in agammaglobulinemic anemic chickens and shortened survival of agammaglobulinemic anemic RBC (Aγ-RBC) in three of four normal chickens. Aγ-RBC could be specifically agglutinated by an antibody present in the normal serum of several species. Several lines of evidence indicate that chicken immunoglobulin is not on the Aγ-RBC surface and hence not involved in the agglutination reactions observed. Infectious agents were not identified in the agammaglobulinemic anemic chickens. The absence of an autoantibody in this hemolytic anemia, which is similar to that described in other species as “autoimmune,” was consistent with the concept that autoantibodies may not play a primary pathogenetic role in at least some phenomena generally characterized as autoimmune. The most likely etiologic factor in the hemolytic anemia was thought to be an unidentified infectious agent, although the contribution of the absence of immunoglobulin per se could not be adequately evaluated. The pertinence of the use of the agammaglobulinemic chicken as a model for “autoimmune” phenomena in immune deficient humans was discussed, and the necessity of proper controls in using anti-immunoglobulin sera in the investigation of hemolytic anemias was stressed.