bluuebunny/crossref_metadata_2025 huggingface dataset:
The Purification and Reactivity of the First Component of Complement from Guinea Pig, Human and Canine Sera
Authors: Tamura Noboru , Nelson Robert A. , Paterson Leslie M. , Haugland Elaine
Published: December 2022

A simple procedure for separation and purification of the first component of guinea pig complement is described. The product has high specific reactivity, low protein content and minimal content of other complement components. The formation of EAC1 using sheep erythrocytes sensitized with purified IgG was greater at 0°C than at 37°C. In contrast, the formation of EAC1 using sheep erythrocytes sensitized with IgM proceeded more effectively at 37°C than at 0°C.

Preliminary results are presented on measurements of C1 by immune adherence as well as by immune hemolysis. Estimates of the number of molecules that are required for a 50% endpoint in each of these two reactions were 2 and 10 molecules per erythrocyte, respectively.

Certain problems concerned with the exact definition of the molecular weight of C1 in native serum are discussed.
A Cytotoxic Antiglobulin Technique for Assay of Antibodies to Histocompatibility Antigens
Authors: Fass Leroy , Herberman Ronald B.
Published: December 2022

A method for increasing the cytotoxic activity of antibodies to lymphoid cells has been developed. Mouse isoantisera with 7 S and 18 S antibody activity were tested against an allogeneic lymphoid tumor cell. A variety of heterologous antiglobulins with different specificites were examined for their ability to produce inceased cytotoxicity. Both anti-IgG and anti-light chain reagents increased the cytotoxic titers of the 7 S antibody as much as 16-fold. Antibodies against mouse light chains were also able to enhance the cytotoxicity of the 18 S antibody.

The degree of enhancement of cytotoxicity was considerably less than that found for antiglobulin reactions with erythrocytes and the possible explanations for these differences were discussed.
Critical Variables of the Jerne Plaque Technique As Applied to Rodent Antibody-Forming Systems Responding to Heterologous Red Cell Antigens
Authors: Hübner K. F. , Gengozian N.
Published: December 2022

Improved conditions were developed for the Jerne plaque technique; a simple test (“hemolytic spot-testing”) allows a study of interspecies antigen-antibody systems and interspecies complement effects as well as ionic requirements for hemolysis in agar. These optimal conditions were applied to the Jerne plaque test with parallel results.

The Jerne technique can easily be applied to study the hemolytic antibody response of C3BF1 mice, Golden Syrian hamsters, and Wistar rats immunized with heterologous RBC. Optimal results depend on the type and dilution of complement used for a certain heterologous combination: guinea pig complement gives best results in the mouse anti-sheep system; rabbit complement is superior in the mouse anti-rat system; and human complement is hemolytically most active in the mouse anti-hamster, hamster anti-mouse, and rat anti-mouse combination.

Each C′ requires different ionic conditions of the reaction medium. It is demonstrated that optimal results of the Jerne plaque test depend on adequate concentrations of Ca++ and Mg++ in the agar. The data suggest also that the anticomplementary effect of agar is ionic. This hemolysis-reducing or inhibiting factor is overcome by DEAE dextran and higher cation concentrations of the agar. The mechanism of action of the polycation and Ca++ and Mg++ cations is discussed.

The hemolytic activity of complement of different species is not only reflected in variation of the number of plaques but also in average plaque size. A quantitative complement-antibody correlation is considered as a possible explanation for this observation, and suggests that this method may provide a model for the study of the dynamics of cell lysis.
Two Mechanisms of Immunologically-Induced Injury to Rabbit Platelets
Authors: Des Prez Roger M. , Bryant Richard E.
Published: December 2022

Platelet injury (release of platelet 5-hydroxytryptamine) that occurs when antigen and antibody are added to non-immune platelet-rich plasma has been compared and contrasted to platelet injury that occurs when preformed immune precipitates are used as the challenge. Platelet injury by large concentrations of immune precipitate is not inhibited by citrate, by anticomplementary concentrations of heparin or sodium chloride, or by prior pepsin digestion of the antibody from which the precipitates are prepared. Prior zymosan adsorption of plasma at 16°C in the presence of Na2H2EDTA inhibits the precipitate-platelet interaction.

Platelet injury produced by soluble antigen and antibody in non-immune platelet-rich plasma is inhibited by citrate, by anticomplementary concentrations of heparin and sodium chloride, and by prior pepsin digestion of the antibody. Prior zymosan adsorption of plasma at 16°C in the presence of Na2H2EDTA is, however, without effect.

It is postulated that platelet injury by immune precipitates in citrated plasma results from phagocytosis of the precipitates by platelets and does not require complement, that platelet injury by soluble antigen and antibody in heparinized plasma is a complement-dependent process, and that platelet injury by immune precipitates in heparinized plasma may involve both mechanisms.
The Fixation of Guinea Pig Complement by γ1 and γ2 Immunoglobulins
Authors: Osler Abraham G. , Oliveira Benedito , Shin Hyun S. , Sandberg Ann L.
Published: December 2022

Several years ago the 7 S guinea pig immunoglobulins were separated into two well-defined electrophoretic classes: the slowly migrating γ2 and the more rapid γ1 (1–3). This achievement led to further studies which demonstrated that the ability to activate the complement (C′) sequence was confined to γ2 immunoglobulins. These antibodies failed to mediate passive cutaneous anaphylaxis (PCA) in the guinea pig. In contrast, the γ1 immunoglobulins were capable of eliciting PCA reactions despite their purported deficiency in C′-fixing potency.

We have re-investigated this question and have confirmed all the previous findings excepting the C′-fixing properties of γ1 type antibodies. These immunoglobulins do interact with guinea pig C′ components, but probably by a different pathway.

Hartley strain guinea pigs were immunized with highly substituted dinitrophenylated bovine γ globulin (4) emulsified in complete Freund's adjuvant as described in (5). Initially, 250 µg of antigen were injected into each of the four toepads.
Protein-Carbohydrate Interaction
Authors: So Lucy L. , Goldstein I. J.
Published: December 2022

Two-dimensional agar gel diffusion and quantitative precipitation studies of concanavalin A with several polysaccharides of varying molecular weights (dextrans, levans and glycogens) demonstrated the effect of polysaccharide molecular size on the concanavalin A-polysaccharide precipitation reaction. The effect of the molecular weight of these polysaccharides was manifested by their band positions in agar gel diffusion tests, i.e., the lower molecular weight species formed precipitation bands closer to the concanavalin A well.

The influence of polysaccharide molecular weight is evident also from quantitative precipitation studies, especially in the region of excess concanavalin A and of excess polysaccharide. Equivalence (or maximal precipitation) was reached earlier with the lower molecular weight polysaccharides; the lower molecular weight polysaccharides were also more effective in inhibiting precipitation in the region of polysaccharide excess.
Physicochemical Properties of Human Reaginic Antibodies
Authors: Ishizaka Kimishige , Ishizaka Teruko
Published: December 2022

Reduction of anti-ragweed γE antibodies in 0.1 M mercaptoethanol for 1 hr followed by alkylation with iodoacetamide resulted in a great decrease of their skin-sensitizing activity, whereas treatment of the antibodies with 0.01 M mercaptoethanol did not have a significant effect. The γE antigenic determinants in the molecules were not impaired by reduction with 0.1 M mercaptoethanol and alkylation. The antigen-binding activity of the antibodies was diminished to about one-third by the treatment. It was also found that the affinity of γE molecules for skin tissues was lessened by the treatment. On a weight basis, the ability of γE to block passive sensitization with reaginic antibodies of a heterologous antigen system was diminished to one-fifth by the treatment. Passive sensitization with the reduced-alkylated γE antibodies did not persist for a long time. The results indicated that the effect of reduction-alkylation on the γE antibodies involve both antibody-combining sites and tissue fixation sites in the molecules. The structural basis of the affinity of γE molecules to target cells was discussed.
Reversal of Post-Thymectomy Wasting in Mice with Immunocompetent Cells: Influence of Histocompatibility Differences
Authors: Stutman O. , Yunis E. J. , Good R. A.
Published: December 2022

Reversal of post-thymectomy wasting syndrome in conventional C3H/Bi mice was achieved by intraperitoneal injection of 200 × 10 spleen cells from adult syngeneic, hemiallogeneic and allogeneic donors sharing the same H2 histocompatibility locus with the host. When cells from allogeneic donors differing at the H2 locus were used, reversal of wasting did not occur and all the animals died with a graft-versus-host reaction.

When neonatally thymectomized F1 hybrids were treated after the onset of the wasting syndrome with spleen cells, only syngeneic cells were effective, while parental or allogeneic cells killed the animals as a result of a graft-versus-host reaction.

The reconstituted animals developed ability to reject skin allografts but were tolerant to skin grafts of donor origin, indicating adoptive immunologic restoration and a chimeric state. The chimeric state was demonstrated using chromosome marked donor cells by high numbers of donor type mitosis in the lymphoid organs of the restored animals.

These findings warn against use of immunologically competent lymphoid cells differing from the host by major histocompatibility factors and encourage matching of host and donor in efforts to reverse wasting and establish competence in thymic deficiency syndromes.
Humoral Immunity in Rodent Malaria
Authors: Diggs Carter L. , Osler Abraham G.
Published: December 2022

Parasitemia in young rats infected with Plasmodium berghei can be monitored by electronic counting of particles in blood lysates. The number of particles so obtained provides an estimate of the number of parasitized cells in the blood sample. The method is highly reproducible as compared to conventional techniques, especially as applied to low levels of parasitemia. The further advantage of rapidity is also inherent in the procedure.
Mechanism of Immunologic Tolerance
Authors: Scott David W. , Waksman Byron H.
Published: December 2022

Specific unresponsiveness to BγG was demonstrated in thymectomized, irradiated Lewis rats restored with washed cells from intact lymphoid organs injected with soluble BγG and incubated for 2 hr in vitro. Both delayed sensitization and antibody formation against BγG were reduced in the recipient; responses to PPD and bovine γA-globulin were not affected. Unresponsiveness to soluble ovalbumin could be induced by use of the same technique. Evidence is presented that this phenomenon is not due to the transfer of either free antigen, with the production of low-zone tolerance, or of cell-associated (membrane-bound) antigen. Since comparable unresponsiveness was not induced by incubating lymphoid cells in suspension with BγG, it is suggested that a processing step may be necessary for the production of tolerance or, alternatively, that the physiologic environment of the lymphoid cells controls their reactivity with antigen.
Studies on the Induction of Immunologic Unresponsiveness
Authors: Golub Edward S. , Weigle William O.
Published: December 2022

The variation in the dose of ultracentrifuged human γ globulin (HGG) needed to produce an unresponsive state in different strains of inbred mice was shown to be the result of differences in the efficiency with which these strains process the trace amount of aggregated material remaining. Although mice from both C57BL/6J and BALB/cJ strains remove the aggregated material from the vascular fluids following injection of HGG, the BALB/cJ mice appear to process this material efficiently, and an immune response rather than an unresponsive state results. Conversely, C57BL/6J mice appear to process the aggregated material inefficiently, thus permitting the induction of unresponsiveness to the non-aggregated (tolerogenic) material. When the trace amount of aggregates was removed by salt fractionation, mice from both strains became unresponsive to small doses of HGG. The tolerogenic, deaggregated HGG was rendered immunogenic by chemical aggregation. The dose of HGG to which mice become unresponsive appears to be controlled by a genetic mechanism involving more than one gene.

C57BL/6J mice readily became unresponsive following injections of deaggregated γG isolated from normal rabbit serum, but made an antibody response to γG (ALG) isolated from the sera of rabbits immunized with mouse thymus cells. Mice made unresponsive to deaggregated RγG did not lose their unresponsive state following injection of ALG.

Within the limits of the test system that was used, no low-dose unresponsive state was observed in C57BL/6J and BALB/cJ mice injected with ultracentrifuged HGG.
Inhibition of Capillary Migration by Antigen-Antibody Complexes
Authors: Spitler Lynn , Huber Heinz , Fudenberg H. Hugh
Published: December 2022

Peritoneal exudate cells from normal guinea pigs were passively sensitized by various antigen-antibody complexes and studied in the capillary migration system. The normal migration of cells from the capillary tubes was inhibited by red cell-IgG complexes but not by red cell-IgM complexes. The degree of inhibition depended on the dilution of antibody used to sensitize the red cells and the ratio of sensitized red cells to peritoneal exudate cells. The addition of keyhole limpet hemocyanin (KLH) to chambers containing normal peritoneal cells in the presence of anti-KLH antiserum also produced inhibition of migration. In these experiments, the inhibition was proportional to the amount of antigen added to the cultures.
Delineation of Two Antigenic Markers, “Non a” and “Non g” Related to the Genetic Antigens of Human γ Globulin
Authors: Natvig Jacob B. , Kunkel Henry G. , Joslin Fenneke G.
Published: December 2022

Two new antigenic markers have been described which supplement the known Gm genetic antigens. The first, “non a,” represents a genetic marker for the γG1 subgroup but occurs in all γG2 and γG3 proteins. The second, “non g,” is a genetic marker for the γG3 subgroup but is found in addition on all γG2 proteins. Rabbit and primate antisera to these determinants were readily produced and measurements were carried out by hemagglutination-inhibition. The two antigens appear to reflect the basic amino acid sequences that were modified as a result of the genetic events leading to Gm (a) and Gm (g), respectively. Several examples were cited where the new markers proved informative. It seems probable that failure to appreciate the strong “non a” antigen has lead to confusion regarding certain genetic antigens reported previously.
Development and Function of the Immunoglobulin-Producing System
Authors: Cain W. A. , Cooper M. D. , Van Alten P. J. , Good R. A.
Published: December 2022

The sera of normal chickens and of chickens bursectomized on the day of hatching contain both IgM and IgG antibodies after two injections of sheep erythrocytes. Chickens bursectomized before hatching either are nonresponders, synthesize IgM antibodies only, or synthesize low levels of both IgM and IgG antibodies. It is concluded that precursors of cells competent to synthesize IgM antibodies are activated through bursal influence prior to the activation of ability to synthesize IgG antibody.

Similar specific immunoglobulin deficiency syndromes in human patients could result from developmental failures occurring at different times.
Occurrence and Properties of Rabbit IgG1 Antibody
Authors: Rodkey L. S. , Freeman M. J.
Published: December 2022

Rabbits given multiple injections of bovine serum albumin in complete Freund's adjuvant produced at least three antigenically distinct immunoglobulins (Ig) that were identified by radioimmunoelectrophoresis using specific antisera. IgG and IgM antibody classes appeared early and simultaneously in the response. An antigenically distinct Ig designated as IgG1, which migrated in the fast γ region, appeared later in the response. The latter Ig was intermediate in anionic binding properties to IgG and IgM, sedimented as a 7 S molecule and was mercaptoethanol-resistant. These properties, the lack of homologous skin-fixing antibody activity, its antigenic dissimilarity to colostral IgA protein, and other characteristics, suggested that IgG1 is a minor but distinct and previously unrecognized class or subclass of rabbit Ig. Differences in the specific hemagglutinating ability of IgG1 and the bulk of the IgG class suggested, moreover, that the IgG1 class has unique functional properties.
Lysozyme and β-Lysin Release Stimulated by Antigen-Antibody Complexes and Bacteria
Authors: Tew John G. , Hess W. M. , Donaldson David M.
Published: December 2022

The concentrations of plasma lysozyme and β-lysin increased several-fold following the intravenous injection of either dead bacteria or antigen-antibody complexes into normal rabbits, or of BSA into immunized rabbits. Histamine and serotonin were not responsible for these alterations. The exposure of whole blood to dead Micrococcus lysodeikticus in vitro also resulted in a significant release of β-lysin and lysozyme into plasma. Similar exposures of peritoneal phagocytes to antigen-antibody complexes or bacteria resulted in the release of lysozyme during phagocytosis. Exposure of β-lysin-rich platelets to either antigen-antibody complexes or micrococci in vitro did not cause a consistent release of β-lysin. The in vivo release of β-lysin was accompanied by a decrease in the number of circulating platelets, a decrease in β-lysin per unit platelet, and extensive morphologic damage of platelets. For these reasons the in vivo results could be attributed to the release of β-lysin from platelets.
Subtypes of Human ABO Blood Groups and Subtype-Specific Antibodies
Authors: Mäkelä O. , Ruoslahti E. , Ehnholm C.
Published: December 2022

We advance an hypothesis which explains the presence of anti-A1 in group A2 individuals and of anti-B1 in group Bv individuals without assuming that A1 and A2 receptors are different or that B1 and Bv receptors are different. Taking into account the known fact that the receptor-density is higher in A1 and A2 erythrocytes, the hypothesis states that anti-A1 antibodies have a low affinity for A receptors and cannot form firm bridges between red cells when they use only one valency per cell. Consequently they do not agglutinate A2 cells, on which the density of receptors is too low. They do agglutinate A1 cells, on which the receptors are closer together. Thus subtype-specific antibodies must be multivalent, and bivalent antibodies such as IgG cannot distinguish between high- and low-receptor-density cells. A corresponding explanation may be valid for subtypes of B and anti-B1.

Our results, as far as they go, support this hypothesis. Although normal anti-A and anti-B isoagglutinins contain a small proportion of IgG molecules, we could not demonstrate any IgG in isoagglutinins specific for subtypes A1 and B1. Anti-A1 reagents could be prepared from the purified IgM fraction of anti-A sera but not from the IgG fractions of the same sera.

Oligosaccharides with blood group A or B specificity strongly inhibited agglutination when the test cells had a low density of receptors (A2B or Bv). They were much poorer inhibitors of agglutination when the test cells had a high density of receptors (A1 or normal B). Macromolecular salivary blood group substances were used as inhibitors in parallel tests. They inhibited strongly regardless of the receptor density on the erythrocytes. The use of A2B rather than A1, and Bv rather than B1, erythrocytes in agglutination-inhibition tests for blood group specific oligosaccharides improved the method by making it more sensitive.

According to the above concept, anti-A1 or A2 individuals may possess an autoantibody directed against their own A receptors. The problems arising from this situation are discussed.
Inhibition of the mixed leukocyte culture reaction (MLC) by pre-incubation of the leukocytes with sucrose
Authors: Rode Harold N. , Gordon Julius
Published: December 2022

When leukocytes of two genetically different donors are cultured together for a period of 5 to 7 days, some of the cells transform and undergo mitosis (1). This reaction, which is referred to as the mixed leukocyte culture reaction (MLC), is probably immunologic in nature (2). The experiments to be described indicate that short-term pre-incubation of human blood leukocytes in sucrose inhibits their reaction in mixed leukocyte culture. The evidence obtained suggests that sucrose exerts its effect by acting on phagocytic cells.

Leukocytes were cultured essentially according to the method of Bain et al. (1) as previously described (3). For incubation with sucrose, freshly drawn and sedimented leukocytes were pelleted by centrifugation (500 × G) and were suspended at 5 to 10 × 106 lymphocytes/ml of sucrose. Incubation was at either 4°C or 37°C for periods of 10 to 120 min. Sucrose2 was used at a concentration of 5% to 35% in 0.02 M phosphate buffer, pH 7.2.
Do Antibody-Forming Cells Circulate in the Blood?
Authors: Roseman Jeffrey M. , Leserman Lee D. , Fitch Frank W. , Rowley Donald A.
Published: December 2022

Criteria have been presented for differentiating hemolytic plaques produced by rat cells into three categories. These categories are determined by the effect on plaque formation of rotenone, a metabolic inhibitor, and papain, a sulfhydryl protease. The first in these categories, “true” plaques, are formed by the release of intracellular antibody from cells requiring an intact cellular respiratory system for such release. The second group, “passive release” plaques, are produced by intracellular antibody from cells which can release this antibody in the presence of metabolic inhibitors. The third, “false” plaques, are formed by antibody which was adherent to cell-platelet aggregates.

Spleen cells from recently immunized rats produced either “true” or “passive release” plaques. All plaques produced by peripheral blood cells appeared to be “false” plaques.
Studies of the Chicken Immune Response
Authors: Keily Sister Danile , Abramoff Peter
Published: December 2022

Young chickens, when bleeding or rupture of the spleen accompanied splenectomy, often regenerated small spleens (splenules) in 4 weeks. These splenules, at 8 weeks, were found to be capable of producing antibody, as tested by the Jerne plaque technique. The bone marrow of such chickens was also functional, although to a less extent. When splenectomy was carried out with no bleeding or rupture, no splenules occurred. In these chickens, the introduction of sheep red blood cells 4 weeks later led to the occurrence of antibody-producing cells in the bone marrow but not in the bursa, thymus, liver, lung or cecal lymph. The serum hemagglutinin titer was always lower in splenectomized chickens than in controls, and always rose more slowely and reached its peak titer later. The IgM fraction appeared sooner than the IgG fraction, but the ratio of the two components for 7 days was similar in all groups. There was no significant difference in time of rejection of skin homografts in control and splenectomized chickens.
Passive Immune Lysis with Chromic Chloride-Treated Erythrocytes
Authors: Perucca Philip J. , Faulk W. Page , Fudenberg H. H.
Published: December 2022

A new method of passive immune lysis is presented. This method involves the coupling of protein antigen to chromic chloride-treated human erythrocytes. The kinetics of CrCl3 requirements, antigen dosage, and incubation are presented. The antigen-coated erythrocytes are lysed by an excess of rabbit complement. Experiments documenting the sensitivity and reproducibility of the test as well as the stability and fragility of CrCl3-treated erythrocytes are presented, and various advantages of the test are discussed.
Presence of L-L Interchain Disulfide Bonds in Reconstituted λG Molecules
Authors: Grey Howard M.
Published: December 2022

When partially reduced and alkylated human γ chains were combined with reduced but not alkylated light chains, the reconstituted reoxidized γG protein possessed 65% of its light chains in the form of L-L disulfide-linked dimers. This suggests that light chains are present in γG molecules in such a position that it is sterically possible for them to come into such close proximity to one another that an L-L disulfide bond can be formed under experimental conditions that do not allow formation of an L-H disulfide bond.
Extracellular Esterases of Streptococci and the Distribution of Specific Antibodies in Human Sera of Various Age Groups
Authors: Stock Aaron H. , Lynn Raymond J.
Published: December 2022

Most of the streptococci representative of Lancefield Groups A through O produced an extracellular carboxylic acid esterase active against β-naphthyl acetate. No esterase activity was found in organisms of Groups D and N. Group K strains were not examined. The esterases could be differentiated into five distinct immunologic types based on reactions of identity or non-identity in immunodiffusion with normal adult human sera. A few strains produced esterases that did not react with the human sera in these immunologic tests.

The distribution of antibodies to the five esterases was determined in the sera of infants, children and adults. Antibodies to the esterase E-IV, obtained from animal Group C strains, were rarely found. Antibodies to the other four esterases, E-I, E-II, E-III, and E-V, were present in the sera of newborns as frequently as they were in adults. Presumably these were antibodies obtained from the mother by placental transfer. Infants produced antibodies to these esterases with increasing frequency after the first year of age. The incidence of antibodies to these esterases in children in the 9- and 10-year-old age groups was similar to that observed in adults. The occurrence of antibodies to esterases E-III and E-V obtained from Group B and Group L streptococci, respectively, is contrary to present epidemiologic data pertaining to human infection with these organisms. A cross-reacting antigenic stimulus has not been demonstrated.
Development of Humoral and Cell-Mediated Immunity to Lymphocytic Choriomeningitis Virus in the Mouse
Authors: Larsen Jørgen Hannover
Published: December 2022

The immune response in the mouse to LCM virus has been studied by measuring the CF-antibody titers and the elimination of viremia in syngeneic tolerant virus-carrier mice after adoptive immunization. This latter phenomenon has previously been shown presumably to be the expression of a specific cell-mediated immunity to the virus.

With regard to the humoral response in the adult mouse, it is shown that CF antibodies are formed by the end of 1 week after primary vaccination with LCM virus. They rapidly increase to titers which adjust to a constant level, presumably for the rest of the animal's life. This constant level is the same in secondarily vaccinated or hyperimmunized mice and is unaltered by a booster dose of virus. This constant immunity is most probably caused by a continuous stimulation from viral foci, especially in the kidneys. After adoptive immunization of syngeneic tolerant virus-carrier mice, lymphoid cells from such immune mice are able to give an enormously enhanced immune response. The CF antibodies from both the immune and the adoptively immunized mice are all 2-ME insensitive, indicating their 7 S character.

It is shown that the cell-mediated immunity to LCM virus after primary vaccination develops almost synchronously with the humoral response. During the first weeks of their development, a clear-cut dissociation of the immune responses may be found.
Antigenic Relationships Among the Coronaviruses of Man and Between Human and Animal Coronaviruses
Authors: McIntosh Kenneth , Kapikian Albert Z. , Hardison Kathleen A. , Hartley Janet W. , Chanock Robert M.
Published: December 2022

Antigenic comparison of seven organ culture-grown human coronavirus strains, one tissue culture-grown human strain (229E), several strains of mouse hepatitis virus (MHV) and one strain of infectious bronchitis virus (IBV) was made. With the use of neutralization, complement fixation and fluorescent antibody tests, it was shown that strain IBV-42 was unrelated to the human and murine viruses and that strain 229E bore little, if any, relation to the murine or other human strains tested. In addition, three of the organ culture grown viruses, two of which have been adapted to growth in suckling mice, were shown to be antigenically related to several strains of MHV. The four remaining human strains possessed heterogeneous and poorly defined antigenic characteristics.
The Detection of Neutralizing Antibody to Lymphocytic Choriomeningitis Virus in Mice
Authors: Hotchin John , Benson Lois , Sikora Edward
Published: December 2022

Neutralization tests were done on a variety of human and animal anti-LCM sera utilizing the mouse f.p. inoculation route, which proved to be more sensitive than the i.c. or i.p. route in detecting antibody. The animal's immune status paralleled its f.p. response and refinement of doubtful results with poorly neutralized sera could be obtained by challenging survivors i.c. to determine the immunity end point. If rapid information was desired the mouse i.p. route combined with assay by weight change measurement proved only slightly less sensitive than the f.p. route. Salt-free bovine serum albumin as a virus and serum diluent increased the sensitivity of the f.p. neutralization test approximately 600 times.

Neutralization was demonstrated following inoculation of virus and antisera in different f.p. of the mouse on the same or on different days, but this method was not as quantitatively sensitive as the inoculation of mixtures. Protection was also noted when the serum was given by the i.p. route and the virus by the f.p. route.

Low levels of antibody were detected in LCM immune mouse sera 20 to 60 days after infection using the SFA diluent-f.p. method. No neutralizing antibody was detected in PTI (LCM neonatally inoculated) mice. High titers of neutralizing antibody were found in mice which had suppressed LCM virus infection several months previously. This reverses the previously held view that mice could not normally develop significant neutralizing LCM antibody, and shows that their response in this respect is normal, but extremely slow. These results are discussed in relation to their bearing on the pathogenesis of slow virus disease.
Isolation and Characterization of Free λ Chain of Immunoglobulin Produced by an Established Cell Line of Human Myeloma Cell Origin
Authors: Matsuoka Y. , Yagi Y. , Moore G. E. , Pressman D.
Published: December 2022

A λ-type light chain of immunoglobulin produced by an established cell line (No. 8226) of human myeloma cells from patient S was isolated in an essentially pure state from spent culture medium. The purified preparation (S-λ-medium) showed the common physical and chemical features of λ-type Bence Jones proteins. At neutral pH, S-λ-medium showed a single peak (S20, w = 3.6) in the ultracentrifuge and existed as a dimer, some of which reverted to the monomer in 1 M proprionic acid. Tryptic peptide maps of S-λ-medium indicated that this λ chain, produced in vitro, was a complete light chain which was composed of both “constant” and “variable” portions.

Antigenic determinants of S-λ-medium were compared with the Bence Jones protein of patient S from whom the cell line had been derived, and with other light chain preparations. In addition to the common λ determinants, S-λ-medium had particular determinants which were found only in the Bence Jones protein of patient S, but not in others. This provides strong evidence that the cell line No. 8226 is the progeny of myeloma cells from patient S.
A Survey of Hemagglutinins in Various Seeds
Authors: Allen Nancy K. , Brilliantine Laura
Published: December 2022

Saline extracts of 2663 varieties of plant and vegetable seeds were tested for human blood group activity. Of these, 1635 were inactive, 711 were nonspecific, 227 hemolyzed red cells of all groups and 90 were specific. Twenty-nine of the 90 specific extracts possessed anti-M and anti-N activity. A new lectin with anti-M specificity derived from the seeds of Iberis amara was discovered. This is the first example of anti-M lectin from a plant source and the first lectin found in this species and family. The remaining 61 specific extracts either had been previously reported or were too weak to be of value.
The Adjuvant Effect of Powdered Ferric Oxide: Enhancement of Response to Mycoplasma Pneumoniae and Respiratory Syncytial Virus Vaccines
Authors: Warren Joel , Kende Meir , Takano Kouichi
Published: December 2022

The ability of powdered magnetic oxide to adsorb respiratory syncytial virus and M. pneumoniae, and methods for simple elution of these agents have been described. When formaldehyde-inactivated vaccines incorporating iron oxide were tested in experimental animals, the metallic salt was found to act as an adjuvant and increased the antibody response. In a comparison with mineral oil-Arlacel adjuvant, it was found that the oil adjuvant was superior, but the Fe2O3 preparations were definitely superior to non-adjuvanted controls. Histologic examination of the site of inoculation revealed a mild, non-inflammatory response consisting of prolonged accumulation of tissue macrophages accompanied by slow dissolution and adsorption of the iron oxide. It is suggested that antigens adsorbed to iron oxide may be directed to various sites by the appropriate application of magnetic force.
Studies on the Mechanism of Immune Adherence
Authors: Nishioka K. , Sekine T. , Okada H. , Mayumi M. , Kawachi S.
Published: December 2022

Sheep erythrocytes sensitized with antibody and carrying on their surface the first four components of complement (EAC′1423) are capable of adhering to human erythrocytes. Removal of C′1 and C′2 from EAC′1423 does not affect the immune adherence reaction (IA).

Treatment of EAC′43 or EAC′4 with pronase results in the removal of C′4 without affecting the antibody. Pronase-treated EAC′43 cells retain IA activity with human erythrocytes.

Tannic acid-treated cells can be coated with highly purified C′3. The resulting C′3-coated sheep erythrocytes adhered to human erythrocytes but not to non-primate erythrocytes. Treatment of C′3-coated cells with C′3 inactivator destroys their ability to adhere to human erythrocytes. Isotope-labeled C′3 on tanned cells was 1/32 as effective in IA as C′3 on EAC′1423.

These results indicate that C′3 is the only component required for the IA reaction with human erythrocytes, and may enable the elucidation of the molecular structure of C′3 in relation to its immune adherence reactivity.
Cleavage Products of Components of Human Complement
Authors: Lepow I. H. , Patrick R. A.
Published: December 2022

Both human C′3 and C′5 may serve as substrates from which biologically active fragments may be cleaved by various enzymatic pathways, including prior acting components of complement and trypsin. The fragments share general properties such as contraction of guinea pig ileum with tachyphylaxis and degranulation of mast cells with histamine release and, therefore, meet criteria for designation as anaphylatoxin. However, the fragments from C′3 (C′3a) and C′5 (C′5a) are distinctly different with respect to physicochemical properties and the species of mast cell on which they act. C′5a corresponds to the classical anaphylatoxin generated by incubation of fresh rat serum with agar, based on cross-desensitization of guinea pig ileum and lack of activity on rat mast cells. C′3a, on the other hand, does not cross-desensitize guinea pig ileum to C′5a or rat agar anaphylatoxin and liberates histamine from rat mast cells. Anaphylatoxin is, therefore, a general term descriptive of more than one molecular entity.
Electrofocusing and Electrophoretical Analysis of Complement Components
Authors: Mukojima T. , Tachibana T.
Published: December 2022

Isoelectric points of all nine components of guinea pig complement were determined by electrofocusing, employing Ampholine ampholites. The following results were obtained (in pH units): C′1 5.3–6.0, C′4 5.90, C′2 5.30, C′3 6.25, C′5 5.10, C′6 6.63, C′7 5.78, C′8 6.34, C′9 5.18 and C′3 inactivator 6.45.

The electrophoretic mobilities of complement components determined by immunolysoelectrophoresis were reported previously. After electrophoresis of a complement component in polyacrylamide gel, hemolytic bands were developed by overlaying an appropriate intermediate complex and complement components in agarose. C′1 showed three distinct hemolytic bands. They were defined as F (fast moving), M (median moving) and S (slow moving) bands. By electrofocusing, three peaks of human C′1 were observed at pH 4.56, 4.95 and 5.40. The electrophoretic mobilities of these C′1 peaks were examined by immunolysoelectrophoresis. The first migrated in the F, the second in the M and the third in the S band.
Comparison of Aa1 Allotypic Specificity Carried by Rabbit IgG and IgM
Authors: Segre Mariangela , Segre Diego , Inman F. P.
Published: December 2022

Sheep red blood cells (RBC) sensitized with a sub-hemolytic dose of anti-sheep RBC IgG of a1, b4 allotypic specificity did not hemolyze upon addition of complement, but hemolyzed upon successive treatment with anti-a1 serum and complement. When sheep RBC so sensitized were incubated in agar together with spleen cells from an a3, b4 rabbit which had been immunized with a1, b4 IgG, hemolytic, complement-dependent plaques appeared around cells secreting anti-a1 antibodies. Plaques were specifically inhibited by Ig bearing the a1 allotypic specificity. Thus, it was possible to compare a1 IgG with a1 IgM for efficiency of plaque inhibition. The molar concentration of IgG required to inhibit 50% of the plaques was about 27-fold smaller than the corresponding concentration of IgM (calculated on the basis of the molecular weight of the subunit), showing that antibody produced by single cells had greater average affinity for the immunizing IgG than for the cross-reacting IgM. As IgM, used in sufficient concentration, caused 100% plaque inhibition, there was no evidence of cells producing antibody specific for a determinant present only on IgG. All experiments were done with a single batch of immune spleen cells, which were preserved in liquid nitrogen.
Inhibition of in vitro Cell Migration in Experimental Allergic Encephalomyelitis
Authors: Rauch Helene C. , Ferraresi R. W. , Raffel S. , Einstein Elizabeth Roboz
Published: December 2022

Peritoneal exudate cells obtained from guinea pigs within 7 days following treatment with a dose of purified basic protein from bovine spinal cord emulsified in complete Freund's adjuvant, which ordinarily induces allergic encephalomyelitis, showed inhibition of migration when cultured in the presence of the encephalitogenic antigen. Since guinea pigs that occasionally survive such an encephalitogenic challenge have sensitive cells, as measured by the in vitro migration-inhibition test, it is suggested that factors in addition to hypersensitive cells may be involved in the onset of disease. In animals pretreated with antigen to avert disease, the development of hypersensitive cells appeared to be substantially inhibited. However, animals in which the appearance of disease was suppressed by treatment after the encephalomyelitis-inducing injection yielded cells which were inhibited in culture.
Inhibition of Macrophage Migration by a Skin-Reactive Polysaccharide from BCG Culture Filtrates
Authors: Godfrey H. P. , Baer Harold , Chaparas S. D.
Published: December 2022

A skin-reactive polysaccharide isolated from unheated BCG culture filtrates (GAE) inhibited guinea pig peritoneal exudate cell migration to the same degree as PPD in guinea pigs sensitized 2 to 3 months previously. The hydrolyzed polysaccharide and a second polysaccharide, both skin non-reactive, were inactive in this in vitro model of delayed hypersensitivity. The degree of migration inhibition correlated well with skin test size in recently sensitized (2 to 3 months) animals for both polysaccharide and protein antigens but bore no relationship to serum antibodies. With time, the ability of exudate cells from skin-reactive animals to react in vitro to the polysaccharide antigen disappeared while in vitro reactivity to PPD remained. We conclude that GAE, as well as a number of other polysaccharides, is able to induce and elicit delayed hypersensitivity.
Ultrastructure of Rosette-Forming Cells in the Mouse during the Antibody Response
Authors: Storb U. , Bauer W. , Storb R. , Fliedner T. M. , Weiser R. S.
Published: December 2022

Ninety-four single rosette-forming (RF) cells obtained from the spleens of mice 2 to 6 days after primary injection of sheep erythrocytes were studied with the electron microscope. The observed RF cells could be classified in five ultrastructurally distinct groups, 1) lymphocytes, 2) plasma cells, 3) “cells with pale nuclei,” 4) “cells with cytoplasmic bodies,” and 5) macrophages.

Two morphologic findings had not been described in antibody-containing cells before. 1) In most of the medium-sized and in all the large RF lymphocytes clusters of ribosomes were surrounded by patches of matrix more electron dense than the rest of the ground substance and less dense than the ribosomes. The significance of these patches with respect to antibody synthesis is discussed. 2) Cells in Group 4 contained round lysosome-like cytoplasmic bodies, which were larger (τ;2 µ diameter) than the lysosomes described in lymphoid cells transformed by mitogens. The cytoplasm of some of these cells was otherwise undifferentiated.

The developmental interrelations of the various RF cells are discussed in terms of morphologic criteria and sequence of appearance.
A Study of the Neutralization of Rhinoviruses by Means of a Plaque-Reduction Test
Authors: Fiala Milan
Published: December 2022

Neutralizing antibodies to four rhinoviruses were measured by the reduction in the number of plaque-forming units of virus by antiserum. Both the 50% plaque-reduction titer and the neutralization constant, K, were used to characterize reproducibly the neutralizing activity of an antiserum. The 95% confidence interval of 50% plaque reduction titers was found to be ±1.48-fold. The sensitivity of the plaque-reduction test exceeded that of the end point tube test. The rate of neutralization of rhinovirus types 2 and 12 was first-order for only a short period after which the rate decreased, indicating the presence in the viral population of a persistent fraction.
Immunochemistry and Genetics of a “New” Allotypic Specificity Ae14 of Rabbit γG Immunoglobulins: Recombination in Somatic Cells
Authors: Dubiski S.
Published: December 2022

A “new” allotypic specificity of rabbit immunoglobulins (A14) is described. The determinant is localized on the Fc portion of heavy chains of rabbit γG. The specificity is inherited in a simple Mendelian manner and the gene controlling its synthesis is localized at the locus Ae, which is distinct from Aa and Ab loci; the product of the gene allelic to the Ae14 gene is not yet known. There is a strong indication that the loci Ae and Aa are closely linked in the germline, but that they may undergo recombination in the somatic cells, thus causing the synthesis of recombinant molecules.
“In vitro” cell reactions in delayed hypersensitivity
Authors: Salvin S. B. , Nishio Jane
Published: December 2022

Delayed hypersensitivity has been characterized primarily by responses “in vivo” in either man or experimental animals. Typically, therefore, the slowness of the indurated response to intradermal injection of antigen, the mononuclear nature of the cellular infiltrate, the lack of correlation between the extent of induration and the amount of antibody, and the passive transfer of hypersensitivity with lymphoid cells and not with antiserum all constitute delayed hypersensitivity “in vivo”. Animals possessing these characteristics may also possess an “in vitro” correlate, namely, the inhibition of migration of lymphoid cells in the presence of specific antigen. Initially, cell migration from fragments of lymphoid organs was inhibited in tissue culture by specific antigen (1–3). This inhibition of migration could not be induced by circulating antibody, and apparently therefore is not associated with conventional circulating globulins (4).

Additional “in vitro” systems for the study of migration inhibition in delayed hypersensitivity have recently been introduced (5, 6).
Identification of IgA in Rat Serum and Secretions
Authors: Nash D. R. , Vaerman J. P. , Bazin H. , Heremans J. F.
Published: December 2022

Although serum and/or secretory IgA has been well defined in the human as well as in several laboratory animals including mice, rabbits and dogs (1, 2), this immunoglobulin has not been adequately identified and characterized in the rat. Several reports have appeared in which a rat serum immunoglobulin of relatively slow mobility was referred to as IgA (3–5) or γ1 (6). This identification was made, for the most part, on the basis of electrophoretic mobility. The purpose of this communication is to demonstrate that the mobility of rat serum IgA is faster than that of the presently known rat immunoglobulins, and that some of the previous papers dealing with rat IgA refer to a serum immunoglobulin which may in fact be homologous to mouse IgG1 (IgF).

Of the many possible criteria which can be used in identifying a specific immunoglobulin class within a species, the most precise would appear to be a comparison of amino acid sequences between known immunoglobulins and the protein being studied.
Serologic Activity of Antigen Isolated from the Body Fluid of Ascaris Suum
Authors: Oliver-González José , Hurlbrink Patricia , Conde Encarnita , Kagan Irving G.
Published: December 2022

An antigen prepared from the body fluid of Ascaris suum by fractionation on Sephadex G-200 and purification by cellulose acetate electrophoresis was evaluated for the diagnosis of visceral larva migrans (VLM) in adults and children. The antigen was highly reactive with sera (95%) from children with VLM. Among the sera of adults from an area endemic for ascariasis, 65% were positive. It is suggested that positive reactions in these cases are due to an antigen present in migratory phases, which is also present in the body fluid of Ascaris. The high incidence of reactions of the antigen with Ascaris antibody may serve as a useful serologic adjunct in the diagnosis of visceral larva migrans in suspected cases among children.
Lysosome Stabilization by Leukocyte Granule Membrane Antiserum
Authors: Persellin Robert H.
Published: December 2022

Antiserum prepared in the rabbit to rat peritoneal leukocyte granule (lysosome) membranes retarded the release of enzyme activities from isolated lysosomes. This stabilizing action, isolated in the γG fraction of antiserum, protected intact rat liver large granules stressed by 37°C incubation and also protected rabbit peritoneal leukocyte granules exposed to Triton X-100. The stabilizing activity could be removed by absorption with homologous and heterologous membranes.

Since lysosomes are implicated in the induction of various forms of tissue damage, a membrane-reactive antibody capable of stabilizing lysosomes could act to retard the inflammatory process.