bluuebunny/crossref_metadata_2025 huggingface dataset:
Human Eosinophil Adherence to Serum-Treated Sepharose: Granule-Associated Enzyme Release and Requirement for Activation of the Alternative Complement Pathway
Authors: Metcalfe Dean D. , Gadek James E. , Raphael Gordon D. , Frank Michael M. , Kaplan Allen P. , Kaliner Michael
Published: December 2022

Human eosinophils, obtained in greater than 90% purity from patients with idiopathic hypereosinophilia, were found to adhere to serum-treated Sepharose beads and to release granule-associated enzymes. The release of aryl-sulfatase, an eosinophil-specific enzyme, was found to occur during a 60-min time period and to be dependent upon the presence of serum or a component of serum coating the Sepharose beads. IgG was examined as a possible opsonic factor and no significant role was evident. The factor in serum responsible for inducing eosinophil adherence and enzyme release was heat-labile and was not generated in C3-deficient serum, but was formed upon reconstitution of C3-deficient serum with purified human C3. The activation of the Sepharose particles by normal human serum was found to be dependent upon the presence of magnesium but not calcium, and to occur normally in C2, C4, C5, and C6-deficient sera, suggesting a requirement for the activation of C by the alternative pathway. With 125I anti-C3b/c, serum-exposed Sepharose was shown to exhibit C3b/c on its surface. Furthermore, human RBC were observed to adhere to serumtreated Sepharose consistent with the presence of C3b on the bead surface. Although the opsonic factor has not been directly identified, the concordance appreciated between the loss of immune adherence and the capacity of serum-treated Sepharose to generate aryl-sulfatase release after heating in 2 M salt as well as after trypsin treatment suggests that both phenomena may be related. Thus, eosinophils encountering a noningestible surface capable of activating the alternative C pathway may adhere to that surface and release granule-associated enzymes.
Interaction of T Cells with Allogeneic Macrophages in Mixed Leukocyte Cultures in Rats
Authors: Oehler J. Ronald , Herberman Ronald B.
Published: December 2022

The role of macrophages in rat mixed leukocyte cultures was investigated. Previous studies indicated that macrophages in the responding cells suppressed the proliferative and cytotoxic responses. When, in the present study, macrophages were depleted from the stimulating cells, the proliferative and cytotoxic responses were greatly depressed. Reconstitution of the macrophage-depleted cultures with low concentrations of irradiated peritoneal exudate cells of the stimulator but not the responder haplotype resulted in the generation of strong cytotoxic responses. Further, highly purified stimulator peritoneal macrophage populations induced the generation of strongly cytotoxic effector cells, which suggested that macrophages are the predominant stimulating cells in rat mixed leukocyte cultures.
Cortisone As a Probe for Cell Interactions in the Generation of Cytotoxic T Cells
Authors: Lee Kwok-Choy
Published: December 2022

The effect of cortisone on the in vitro cell-mediated cytotoxic response was investigated with respect to the participating cell types, viz., the cytotoxic T cell precursor (CTP), the helper cell, and the accessory (A) cell. Spleen and thymus cells from cortisone-treated CBA/CaJ mice were unable to generate alloreactive cytotoxic T cells when cultured with semiallogeneic (CBA/CaJ × DBA/2JF1) spleen cells or allogeneic (P815) tumor cells. This immunosuppression could be reversed by the addition of 2-Me to the cultures, which suggests that the CTP and helper cells were present but unresponsive. Measurements of the CTP frequency and helper activity in cultures containing 2-Me confirmed this possibility. Thus, limiting dilution analysis of spleen and thymus cells from cortisone-treated mice showed that they contained higher frequencies of CTP than cells from normal mice. Helper cell activity, as measured by the secretion of helper factor by primed cells, was found to be higher after cortisone treatment. The possibility of a defect in A cell function in spleen cells of cortisone-treated mice was suggested by 1) the ability of irradiated peritoneal macrophages to reconstitute the response, and 2) the greatly reduced efficiency of irradiated spleen cells (adherent or nonadherent) from cortisone-treated mice to restore responsiveness to nonadherent, A cell-deficient normal spleen cells. Although cortisone may exert its immunosuppressive effect on helper cells and CTP directly, it is likely that the deficiency in A cell function is at least partially responsible for the unresponsiveness of cortisone-treated cells.
Cellular Events in Tolerance
Authors: Venkataraman M. , Scott David W.
Published: December 2022

The numbers and fate of antigen-binding cells (ABC) in neonatal and adult mice rendered tolerant to fluorescein (FL)-labeled heterologous γ-globulins were studied. Similar numbers of FL-ABC were observed 1 day after tolerogen in both adult and neonatal mouse spleens; by 7 days after tolerization, no FL-ABC were observed in either case. Reinjection with FL-tolerogen at 7 days led to the detection of normal numbers of ABC in adult mice but significantly reduced numbers in neonates. This suggests that neonatal ABC either have been deleted or have failed to resynthesize surface receptors.

Two weeks after tolerance induction, spleen cells from these tolerant mice were cultured with Escherichia coli lipopolysaccharide (LPS), a polyclonal B cell mitogen, or with specific antigen. Tolerant adult spleen cells made an equivalent anti-FL response to that of the uninjected controls when stimulated with LPS, but were unresponsive to specific antigenic triggering. In contrast, spleen cells from neonatally tolerized mice were unresponsive to either specific or nonspecific (LPS) stimulation. Thus, these neonatally tolerized spleen cells lose sensitivity to polyclonal-stimulating agents (along with their receptors), or more simply, are deleted.
A New Method for Evaluation of Intracellular Inhibition of Multiplication or Killing by Mononuclear Phagocytes
Authors: McLeod Rima , Remington Jack S.
Published: December 2022

A new method for evaluation of the capacity of mononuclear phagocytes to inhibit intracellular microorganisms is described. The system provides a means for assessing this effect of macrophages without concern for multiplication of extracellular organisms, effect of antibiotics, and the potential observer bias which may result from visual evaluation. It involves measurement of amount of [6-3H]UdR incorporated by Toxoplasma gondii. Differences between uptake of [6-3H]UdR by infected and uninfected macrophages can be augmented by cytosine arabinoside as this agent inhibits macrophage DNA synthesis but does not substantially alter DNA synthesis by the test organism, T. gondii.
Genetic Analysis of Generation of Serum Interferon by Bacterial Lipopolysaccharide
Authors: Apte Ron N. , Ascher Orna , Pluznik Dov H.
Published: December 2022

Administration of bacterial lipopolysaccharides (LPS) to mice causes interferon (IF) to appear in their blood stream. The generation of this serum IF is “early” and its maximum concentration appears 2 hr after an inoculation of a single optimal dose of LPS (100 µg). Two inbred strains of mice differing in their ability to generate LPS-induced serum IF were used: a high responder strain (C3H/eB), which generates high levels of IF (about 100 IF units/ml serum), and a low responder strain (C3H/HeJ), which fails to generate any detectable amounts of IF in the serum. A genetic analysis was carried out and circulating IF production was determined in the F1 hybrid, in the F2 generation, and in the progeny of backcrosses of F1 hybrids to either parent strain. The results indicate that a single dominant autosomal gene is responsible for the difference in circulating IF production between both parent strains. In addition it was shown that this type of IF is stable to treatment at pH 2 for periods of time as long as 48 hr, but is labile to heating at 56°C for 30 min.
Histocompatibility-2 System in Wild Mice
Authors: Zaleska-Rutczynska Zofia , Klein Jan
Published: December 2022

Sixteen B10.W congenic lines carrying on C57BL/10Sn or B10 background H-2 haplotypes extracted from wild mice are described. The lines were tested serologically in two ways: first by antisera against known H-2K and H-2D private antigens of inbred strains, and second by antisera made in inbred strains against antigens carried by the B10.W lines. Both direct cytotoxicity and absorption tests were used. The analysis resulted in the serologic identification of the H-2 haplotypes carried by the 16 B10.W lines. Lines B10.STA10 and B10.STA12 are serologically indistinguishable; so are lines B10.KPA42, B10.KPA132, and B10.SNA57, whose H-2 haplotypes resemble the H-2v haplotype of B10.SM; all other B10.W lines are different from one another and different from all inbred strains known. In total, 10 new H-2 haplotypes (H-2w11 through H-2w20) and 16 new H-2 antigens (H-2.110 through H-2.125) were identified. One haplotype was discovered carrying new combination of known H-2 antigens: H-2w19, which is H-2K.19, H-2D.2. Known H-2 antigens in combination with new one were discovered in haplotypes H-2w8 (H-2.23 and 110), H-2w12 (H-2.23 and 112), and H-2w11 (H-2.26 and H-2). The first two of these three haplotypes are probably the result of intra-H-2 crossing over that occurred during the production of B10.W lines; the last one (and the H-2w19 haplotype) is probably a natural recombinant.
The First Complement Component: Evidence for an Equilibrium between C1s Free in Serum and C1s Bound in the C1 Complex
Authors: Bartholomew Richard M. , Esser Alfred F.
Published: December 2022

An equilibrium between free C1s and C1s bound in macromolecular C1 exists in human serum. This equilibrium can be utilized to incorporate radioiodinated C1s into serum C1. Human sera were incubated for 40 hr at 4°C with 125I-C1s to allow the exchange between free and bound C1s to reach equilibrium. The C1 complex labeled in this manner was separated from the majority of serum proteins by centrifugation in linear 10 to 30% sucrose density gradients. The resulting fractions containing 125I-C1 can be used directly and conveniently in C1 activation assays that detect the cleavage of proenzyme C1s. Electrophoretic analysis on polyacrylamide gels showed the presence of only proenzyme 125I-C1s in serum C1, whether or not the applied labeled material contained 125I-C1s̄ or other labeled proteins. The inability of C1 to incorporate C1s̄ was shown to be the result of decreased stability of C1 upon activation.
Murine B Lymphocyte Heterogeneity: Distribution of Complement Receptor-Bearing and Minor Lymphocyte-Stimulating B Lymphocytes among Cells with Different Densities of Total Surface Ig and IgM
Authors: Scher Irwin , Ahmed Aftab , Sharrow Susan O.
Published: December 2022

The distribution of complement receptor-bearing (CR+) and minor lymphocyte-stimulating (Mls)-defined lymphocyte-activating determinants (LAD) among B lymphocytes with different densities of total surface immunoglobulin (sIg) and sIgM was determined. B lymphocytes that bore intermediate densities of sIg and low densities of sIgM had the highest frequency of CR+ cells and were the most active in expressing Mls-defined LAD. The distribution of these and other surface markers on B-lymphocyte subpopulations is discussed.
Changes of Fcγ Receptor-Related Properties Induced by Interaction of Human Lymphocytes with Insoluble Immune Complexes
Authors: Cordier Geneviève , Samarut Christiane , Revillard Jean-Pierre
Published: December 2022

An in vitro model was designed to investigate the fate of membrane receptors for Fcγ (FcγR) on human lymphocytes after a first interaction with antigen-antibody complexes or aggregated immunoglobulins. After the binding of erythrocyte-antibody (EA) complexes to FcγR, a process of dissociation occurs. It is temperature dependent and faster at 37°C than at 4°C. In addition, the dissociation becomes progressively irreversible as shown by the inability of dissociated cells to reform EA rosettes or to act as effectors in antibody-dependent cell cytotoxicity. Similarly, inhibition of EA rosette formation by aggregated human IgG could be reversed by trypsinization after incubation at 4°C but not at 37°C. Several mechanisms that might have accounted for the irreversible dissociation of EA rosettes could be excluded: no antibody fragments could be evidenced at the cell surface, trypsinization did not restore the capacity to form EA rosettes, and lymphocytes were shown to be alive, to incorporate amino acids, and to release β2 microglobulin. The phenomenon was therefore termed modulation of FcγR. Finally, FcγR was not detectable at the cell surface after culture for up to 3 days, suggesting that FcγR was rapidly shed and/or slowly resynthetized after modulation.
Local Transfer of Delayed Hypersensitivity and Cutaneous Basophil Hypersensitivity
Authors: Stashenko Philip P. , Bhan Atul K. , Schlossman Stuart F. , McCluskey Robert T.
Published: December 2022

We have studied the capacity of unfractionated cells, or of purified populations of T and B cells, prepared from the lymph nodes of sensitized guinea pigs with classical delayed hypersensitivity (DH) or with cutaneous basophil hypersensitivity (CBH), to induce reactions in normal syngeneic guines pigs after intradermal injection with antigen (ovalbumin, DNP-BGG, or α,DNP-Lysine11).

Injections of cells or antigen alone produced little or no reaction. Injections of antigen and cells from DH donors resulted in delayed onset reactions, which were indurated and which usually contained only a few basophils. With cells from CBH donors, delayed onset reactions were also produced, but these were less indurated and the infiltrate generally contained higher percentages of basophils. Thus, the reactions resembled those in actively immunized animals.

With cells from DH donors, reactions were consistently produced with unfractionated cells or T cell preparations, with cell doses as low as 0.5 to 1 × 106. B cell preparations also produced reactions, but only with higher cell doses (2 to 3 × 106); these reactions were probably due to contaminating T cells.

With cells from CBH donors, reactions were consistently produced with unfractionated cells, T or B cell preparations in the dose range of 2 to 3 × 106 cells. There was no lower dose at which reactions resulting from B cell preparations could be eliminated while still retaining T cell reactivity. Moreover, doubly purified B cell preparations, with 98% Ig-positive cells, were as effective as singly purified preparations (95% Ig positive). These findings indicate that both T and B cells can initiate CBH reactions and support the interpretation that CBH reactivity is heterogeneous.
A Cross-Reaction between β2-Microglobulin and κ-Light Chains
Authors: Gottlieb A. B. , Engelhard M. , Kunkel H. G. , Tanigaki N. , Pressman D.
Published: December 2022

Certain antisera to immunoglobulins containing κ-chains show the presence of antibodies that cross-react with β2-microglobulin. This was most apparent with an antiserum made to highly purified F(ab) fragments of Fr II γ-globulin. These cross-reactive antibodies caused positive fluorescence and cytotoxicity reactions with a variety of cell types including T cells. These reactions were completely removed by absorption with highly purified κ-chains but not with λ-chains or λ immunoglobulins. β2-microglobulin preparations also absorbed or inhibited the special cellular reactivities. Evidence was obtained that HLA-bound β2-microglobulin was more efficient in this respect. The possibility is discussed that similar cross-reactive antibodies may have been involved in some previous studies of inhibition of T cell function by immunoglobulin antisera.
Linkage of Guinea Pig Bf and C4 to the GPLA
Authors: Krönke Martin , Hadding Ulrich , Geczy Andrew F. , de Weck Alain L. , Bitter-Suermann Dieter
Published: December 2022

Inbred and partially inbred strains of guinea pigs were typed for their polymorphic Bf and C4 patterns. In crosses and backcrosses of inbred strains of guinea pigs the inheritance of Bf, C4, and GPLA phenotypes was studied. The presented data indicate linkage of the structural Bf and C4 loci to the major histocompatibility complex (MHC) of the guinea pig, the GPLA system. Evidence for a GPLA-B-Ia crossover was obtained. Bf and C4 typing contributes to the characterization of the GPLA region in some inbred strains thus providing a simple method for determining the state of inbreeding on the one hand and providing the necessary third or fourth region within the GPLA for the detection of informative meiosis, on the other.
Mechanism of Histamine Release by Formyl Methionine-Containing Peptides
Authors: Siraganian Reuben P. , Hook William A.
Published: December 2022

Small, acylated, methionine-containing peptides release histamine from human basophils. The characteristics of this reaction were compared to that of C5a- and IgE-induced release. fMet peptide-induced release requires Ca++ and is inhibited by EDTA in a manner similar to IgE- and C5a-mediated reactions. The fMet-Phe-Met-initiated reaction is complete within 2 min at temperatures of 25, 30, and 37°C; but does not occur at 0°C. There was a large variation in the capacity of leukocytes from different donors to release histamine with fMet peptides. However, there was no correlation in the capacity of leukocytes to release histamine with fMet-Phe-Met and their release with C5a or anti-IgE. The release by fMet-Phe-Met (but not by C5a or anti-IgE) was reversibly inhibited by a nonreleasing tripeptide. Leukocytes could be desensitized to the action of active fMet-peptide by preincubation with the peptide in the absence of cations. After washing, these cells released normally with C5a or anti-IgE. Conversely, cells desensitized to the action of C5a- or IgE-mediated reactions released normally with fMet peptides. There was cross-desensitization between different active peptides, and inactive peptides could not desensitize the leukocytes. Pharmacologic agents had similar effects on C5a and fMet peptide-induced release (e.g., lack of enhancement with deuterium oxide; enhancement with cytochalasin B; and inhibition with aminophylline and dibutyryl cyclic AMP). Therefore, histamine release with fMet peptides is initiated by their binding to and activation of a specific receptor on the basophil; the reaction beyond that point is similar to the C5a-mediated reaction.
Immunoregulatory Properties of Human Plasma in Very Low Density Lipoproteins
Authors: Chisari Francis V.
Published: December 2022

Physiologic concentrations of VLDL isolated from human plasma by ultracentrifugal flotation and molecular exclusion chromatography inhibit the induction of DNA synthesis in human PBM by a variety of mitogenic and allogeneic cell stimuli. Rapid inhibition of PHA responsiveness during a brief (1 hr) VLDL pulse suggests a high affinity interaction between the inhibitor and its target cell(s) with rapid translation of the inhibitory message to intracellular sites responsible for the initiation of DNA synthesis. PBM refractoriness to VLDL when administered later than 6 to 12 hr after PHA suggests that early, inductive events in the DNA synthesis response of PBM of mitogens are susceptible to modulation by this inhibitor. Currently demonstrated inhibition of protein synthesis and previously documented inhibition of sterol metabolism by VLDL suggest that suppression of the DNA synthetic response of PBM might be mediated by one or both of these early prereplicative events.

The VLDL employed in these studies were shown to be free of the previously described inhibitory species of LDL (LDL-In) and were characterized by a variety of physicochemical, biochemical, and immunochemical parameters virtually incompatible with the other inhibitors of PBM DNA synthesis known to occur in human plasma. Considerable presumptive evidence suggests that inhibition depends on the biologic expression of the apolipopeptide B moiety which VLDL share with the bioregulatory low density lipoproteins and that VLDL may represent the physiologic progenitor of these molecules.

The current studies clearly add VLDL to the enlarging family of immunoregulatory lipoproteins biologically active as subphysiologic concentrations and therefore potentially important in the continuing maintenance of immunologic homeostasis. The precise role played by these molecules in the preservation of human immunologic integrity remains to be determined.
The Effects of Neuraminidase and Galactose Oxidase on Murine Lymphocytes
Authors: Kuppers Rudolf C. , Henney Christopher S.
Published: December 2022

The sequential treatment of normal C57BL/6 mouse spleen cell populations with neuraminidase (NA) and galactose oxidase (GO) resulted in cell proliferation, but not in the differentiation of cytotoxic T cells. In contrast, C57BL/6 spleen cells derived from animals primed 5 to 8 months earlier with alloantigen (P815 mastocytoma cells of the DBA/2 strain) both proliferated and demonstrated T cell-mediated cytotoxicity after NAGO stimulation.

T cells differentiating into cytotoxic cells after NAGO treatment demonstrated properties similar to alloantigen-specific ‘memory’ T cells. These were: 1) cytotoxicity developed only from ‘primed’ cell populations, 2) cytotoxicity developed within 24 hr after NAGO treatment, 3) DNA synthesis was not required for the differentiation of cytotoxic cells during the first 24 hr of culture but both DNA synthesis and cell proliferation were required for the cytotoxicity developing after 24 hr, and 4) all cytotoxicity induced by NAGO showed specificity for the priming alloantigen.

It was found, furthermore, that cytotoxicity could be induced at much lower GO concentrations than needed for increased DNA synthesis. We interpret this finding as an indication that NAGO can differentially deliver two ‘signals’ to T lymphocytes: one leading to cell proliferation, the other causing the differentiation of memory T cells into cytotoxic effectors.
Intravascular Aggregation and Pulmonary Sequestration of Platelets during IgE-Induced Systemic Anaphylaxis in the Rabbit: Abrogation of Lethal Anaphylactic Shock by Platelet Depletion
Authors: Pinckard R. Neal , Halonen Marilyn , Palmer John D. , Butler Cooley , Shaw James O. , Henson Peter M.
Published: December 2022

Significant alterations in the circulatory properties of platelets have been documented during IgE-induced systemic anaphylactic shock in the rabbit. Within 30 to 60 sec after i.v. antigen challenge, platelet aggregation occurs in both the venous and arterial circulations. The platelet aggregates then sequester in small blood vessels of various organs, particularly in the lung. The organ sequestration results in the development of a profound thrombocytopenia within 3 to 5 min after antigen challenge. Fifteen min later deaggregation of platelets occurs and the platelets return to the peripheral circulation within normal, prechallenge levels by 60 min. Additional experiments demonstrated that platelet depletion before antigen challenge abrogates the lethal effects and significantly reduces the pathophysiologic manifestations of IgE-induced systemic anaphylaxis. We conclude that the IgE-induced platelet alterations, probably induced by the intravascular release of basophil and perhaps mast cell-derived platelet-activating factor (PAF), play a major role in the pathogenesis of systemic anaphylaxis in the rabbit.
Evidence That C1t (Amyloid P-Component) Is Not a Subcomponent of the First Component of Complement (C1)
Authors: Painter R. H.
Published: December 2022

The recent observation that plasma P-component/C1t binds to agarose in a calcium-dependent fashion provided an opportunity to demonstrate that specific and selective removal of this protein from serum had no effect on the hemolytic or molecular properties of serum C1 activity. Thus, it is concluded that this protein does not form part of the C1 macromolecular complex.
Binding of F(ab′)2 of Normal Human IgG to Human Lymphocytes
Authors: Hofman Florence , Cohen Ada , Zipori Dov , Burstein Rimona , Benich Zvi , Haimovich Joseph
Published: December 2022

The presence of immunoglobulins (Ig) on the surface of B lymphocytes in humans is well established. However, there has been considerable controversy concerning the proportion of lymphocytes in the peripheral blood bearing on their surface Ig of the IgG class. These different results may be due to variations in techniques, potency, and specificity of the reagents or to the presence of various populations of cells displaying surface IgG under particular conditions. The IgG-bearing cell population probably consists of cells possessing endogenously synthesized IgG present on the membrane and cells having Ig adsorbed onto their surface. Most of the IgG binding has been ascribed to the presence of Fc receptors on the cell surface (1). We have found, however, that part of the IgG bound to cells may be attributed to molecules bound through their Fab part.

Materials and Methods. Antigens.
Heterophile Antigen in Bacteria
Authors: Powell H. M.
Published: December 2022

One of 12 strains of Bacterium lepisepticum was found to contain heterophile antigen. Three strains of the hemorrhagic septicemia group of organisms isolated from guinea pigs did not contain heterophile antigen. One strain of B. avisepticus and three strains of B. bronchisepticus did not contain heterophile antigen.
Studies in Hypersensitiveness
Authors: Fineman A. H.
Published: December 2022

The degree of sensitivity of asthmatics is best determined by skin-testing with dilutions of extracts of increasing strength, and apparently not by the reagin content of the blood; this point requires further study. Asthma patients sensitive to rabbit hair, horse dander, or cottonseed, can be classified, by the intradermal or scratch method, into three arbitrary groups, A, B, and C, depending upon the degree of skin sensitivity. The number of constitutional reactions, resulting from intradermal diagnostic tests or specific treatment, will be appreciably reduced if the degree of sensitivity is predetermined.
The Bactericidal Action of the Serum Following Injections of Adrenalin
Authors: Smith Geo. H.
Published: December 2022

Attempts to associate the glands of internal secretion, or the products of these glands, with immunity or with immunological phenomena have not been wanting, the tissue to receive the most attention from this point of view being the adrenal gland. In the literature of physiology as well as in that of immunology are to be found many contributions bearing upon the relationship of the adrenal glands or of adrenalin to questions of resistance and antibody reactions.

Most of the studies which have been made upon this problem have approached the subject from one of three angles: first, the effect of adrenalectomy upon resistance or upon the capacity of an animal to elaborate antibodies; second, the effect of adrenalin as a detoxicating agent and the relationship of the adrenal glands to detoxicating phenomena; and third, the effect of the administration of adrenalin upon the capacity of the body to produce antibodies to artificially administered heterologous antigens.
Immunity to Pneumococcus Produced in Rats by Feeding Tissues of Animals Killed by the Same Organism
Authors: Ross Victor
Published: December 2022

Rats fed on tissues of animals killed by pneumococcus Type I show an increased resistance to intraperitoneal injections of the same germ. The protection seems to be type specific. Protective substances exist in the sera of the immunized animals. Reference is made to the very favorable immunization created by feeding pneumococci. An explanation is suggested for the fact that those types of pneumococci most commonly found in normal mouths are the cause of a minority of actual cases of lobar pneumonia.
On the Value of the Blood-Group Feature As a Means of Determining Racial Relationship
Authors: Grove Ella F.
Published: December 2022

The blood-group feature of the Hirschfelds and of Ottenberg has been studied in the Ainu and in some Filipino tribes. The blood-group feature of the Ainu in one district (Piratori) is very different from that of Ainu living in other districts, the former resembling the blood group feature of some Koreans, the latter resembling that of some Malay peoples. The Sulu Moros and the Samal Moros are both Malayan people resembling each other in their outward physical characters, yet they differ widely with respect to the blood group feature. The blood group feature of the former is the same as that of the Annamese Malays, but that of the Samals is very like the blood group feature of the Hindus. These findings as well as similarly discordant ones that have been reported by others reveal a serious limitation in the use of the Hirschfeld-Ottenberg method of characterizing peoples.
Bacterial Variation
Authors: Edwards Philip R. , Rettger Leo F.
Published: December 2022

The study of the antigenic characters of bacteria has progressed rapidly within the last few years. Investigators have realized that in working with whole cultures of organisms they were dealing with a mass of varied and complex characters.

Andrewes (1922) was the first to observe two distinct antigens in the Salmonella group. By plating his stock cultures he was able to obtain two entities. The cultures arising from certain colonies he found to be specific and incapable of reacting with any immune sera other than those prepared from cultures of the homologous type. The second antigen, found to be present in an almost pure state in other colonies, was non-specific and reacted with all of the types of the group, agglutinating as readily and in as high dilutions with heterologous types as with the homologous.

Andrewes was able to prepare specific or non-specific sera with these strains, depending upon the particular substrains used.
Bacterial Protein-Free Bacteriophage Prepared by Tryptic Digestion
Authors: Arnold Lloyd , Weiss Emil
Published: December 2022

The antigenic bacterial proteins can be removed from bacteriophage by digesting with trypsin.

Bacteriophage is not damaged by tryptic digestion.

Bacterial protein-free trypsinized phage injected into rabbits does not increase their resistance to a lethal dose of the homologous bacteria used in the propagation of the phage.

The only antibodies produced, when bacteriophage is used as an antigen, are antilytic or neutralizing antibodies.
On a New Serum Test in the Diagnosis of Tuberculosis and Syphilis
Authors: Kabelik J.
Published: December 2022

It is quite generally accepted at present that the defense of the body against tubercle infection is cellular and that serum antibodies in this respect are not of any great importance. There are antibodies, however, in the serum and they have been shown to be of real value in the diagnosis of tuberculosis, a fact which the evidence of recent years has fully established.

The serum test is a valuable addition to our diagnostic resources meeting, as it does, a deeply felt want. As is known, the tuberculin test is too sensitive since it indicates that infection, at some time, has taken place. On the other hand the positive recognition of tubercle bacilli indicates an advanced stage of the disease, namely caseation of tubercular foci and hence open phthisis. In this stage the diagnosis of pulmonary tuberculosis is readily made but, as Macchiavelli rightly said, it is then very difficult to cure.
Neutralization of Atopic Reagins in Vivo
Authors: Clarke J. A. , Gallagher May G.
Published: December 2022

“Atopic reagin” is the name given by Coca to the specific, reacting body discovered by Prausnitz and Küstner in the blood serum of atopic individuals. These reagins have been studied by a number of investigators (1, 2, 3, 4) and their existence thoroughly proved. Briefly, they can be demonstrated by injecting the blood serum from a person suffering from asthma (or an allied condition) into the skin of a normal individual. At the site of this injection, atopens or substances which gave a positive skin reaction in the skin of the donor of the serum will also give a positive reaction in the normal skin of the recipient but will give a negative reaction at any other site not previously injected.

Coca and Grove (3) found that, in a mixture of appropriate atopen and reagin containing serum, the reagin was neutralized. That is they were unable to sensitize a normal skin with such a mixture.
Immunologic Release in the Rat Peritoneal Cavity of Lipid Chemotactic and Chemokinetic Factors for Polymorphonuclear Leukocytes
Authors: Valone Frank H. , Goetzl Edward J.
Published: December 2022

Antigen challenge of the rat peritoneal cavity after passive preparation with IgGa-rich hyperimmune antiserum releases lipids that enhance the migration of human polymorphonuclear (PMN) leukocytes in a modified Boyden micropore filter assay. Deproteinated peritoneal fluid was applied to an Amberlite XAD-8 column equilibrated in distilled water and sequential elution yielded 30% of the migration-enhancing activity in water and 70% of the activity in 80% ethanol. The lipids in the 80% ethanol eluate were chromatographed on silicic acid that was developed sequentially with hexane, dichloromethane, acetone, 1-propanol, and ethanol:ammonia:water (6:3:1, v:v:v). The lipids with migration-enhancing activity eluted in hexane and acetone in two discrete peaks. These were separated from the other lipid mediators slow-reacting substance of anaphylaxis (SRS-A) and platelet-activating factor (PAF), which eluted in ethanol:ammonia:water. The activity in the acetone fraction had the functional characteristics of a chemotactic factor in that it required a concentration gradient to stimulate PMN leukocyte migration and induced chemotactic deactivation, whereas the hexane fraction contained a chemokinetic factor that enhanced PMN leukocyte migration irrespective of a concentration gradient. Both lipid factors were preferentially active on neutrophil and eosinophil PMN leukocytes as compared to mononuclear leukocytes. Reversed immunologic challenge of the rat peritoneal cavity with rabbit anti-rat F(ab′)2 antiserum released both the chemotactic and chemokinetic factors, whereas challenge with rabbit anti-rat myeloma IgE antiserum released predominantly the chemotactic factor. Pretreatment of the rat peritoneal cavity with 5,8,11,14-eicosatetraynoic acid (TYA) before direct immunologic challenge inhibited the release of both lipid factors and indomethacin pretreatment suppressed the immunologic release of the chemotactic factor, but not the chemokinetic factor. Thus, immunologic challenge of the rat peritoneal cavity by pathways dependent on antibodies other than IgE generates both PMN leukocyte chemotactic and chemokinetic activities from arachidonate or related fatty acids, whereas IgE-dependent stimulation releases predominantly the lipid chemotactic factor.
Tolerance and Contact Sensitivity to DNFB in Mice
Authors: Moorhead John W.
Published: December 2022

Contact hypersensitivity (CS) to 2,4-dinitrofluorobenzene (DNFB) in mice is a form of delayed hypersensitivity that can be measured both in vivo (ear swelling) and in vitro (antigen-induced proliferation). Experiments were done to determine whether in vivo and in vitro manifestations of the DH reaction were mediated by different T cell subpopulations. DNFB-immune LN cells were separated on nylon wool columns into nonadherent (T-enriched) and adherent (B-enriched) populations and tested for their ability to transfer CS to normal recipients and to respond to antigen stimulation in vitro. Unseparated LN cells transferred CS and gave a strong in vitro proliferative response to the antigen 2,4-dinitrobenzene sulfonate (DNBS). T-enriched LN cells also transferred CS but did not respond in vitro to either PHA or DNBS, although the response to PHA, but not antigen, could be restored by addition of peritoneal macrophages. B-enriched LN cells did not transfer CS, although they gave a strong antigen response in vitro which was T dependent. Additional experiments using anti-Ia serum plus C showed that the T cells that transferred CS were Ia-, whereas the T cells that responded to antigen stimulation in vitro were Ia+. Collectively, these results indicate that distinct T cell subpopulations mediate the in vivo and in vitro counterparts of this DH reaction. The two populations are separable by virtue of one adhering to nylon wool and by differential sensitivity to lysis by anti-Ia serum plus C. These findings are discussed in terms of the role of lymphocyte subpopulations in CS and with regard to in vitro methods used to investigate cellular and genetic requirements for T cell activation.
B Cell Differentiation Induced by Lipopolysaccharide
Authors: Kearney John F. , Klein Jan , Bockman Dale E. , Cooper Max D. , Lawton Alexander R.
Published: December 2022

The mechanisms by which anti-µ chain antibodies suppress LPS-induced differentiation of murine B lymphocytes to plasma cells were examined as a function of age. Concentrations of anti-µ, sufficient to inhibit completely differentiation of adult B lymphocytes paradoxically enhanced cellular proliferation of LPS-stimulated cultures, whereas newborn cells, which are suppressed by much lower concentrations of anti-µ, failed to proliferate. Enhancement of proliferation of adult cultures was dependent upon both the concentration of anti-µ antibodies and the time they remained in culture. Proliferating lymphoblasts from anti-µ-treated cultures expressed H-2 and Ia alloantigens and receptors for Fc (IgG) and C3; they lacked rough endoplasmic reticulum and other ultrastructural characteristics of plasma cells and had no detectable membrane immunoglobulin. Small deposits of mouse IgM were identified in the perinuclear and Golgi regions in 95% of the cells. Cells recovered from suppressed newborn cultures lacked all of these characteristics. A population of cells, stimulated by LPS in the presence of anti-µ, appeared in the spleen between birth and 3 days, and, by 12 days of age, 35% of cells in suppressed cultures had Golgi-associated IgM. These results suggest that anti-µ inhibition of Ig synthesis in LPS-stimulated adult cells is highly specific, occurs subsequent to cross-linkage of surface IgM, and is mediated in intracellular sites by complexes of anti-µ chain antibody and IgM. Similar treatment of newborn spleen cultures results in elimination of immature B cells. These observations relate to the mechanisms of antigen-induced clonal elimination among immature B cells, as well as to the phenomenon of B cell tolerance and regulation of B cell function by anti-idiotypic antibodies in adults.
Phenotypes of the Major Histocompatibility Complex in Wild Rats of Different Geographic Origins
Authors: Cramer Donald V. , Davis Bridgett K. , Shonnard John W. , Štark Otakar , Gill Thomas J.
Published: December 2022

The serologically detectable (Ag-B, H-1) histocompatibility antigens and mixed lymphocyte responses of geographically widespread populations of wild rats (Rattus norvegicus) were examined with reagents and cells derived from standard inbred rat strains. The histocompatibility antigens of the vast majority of wild rats from France, England, and the United States were very similar, if not identical, to those present in the inbred strains. All but one out of 89 animals displayed one or two Ag-B specificities detected by operationally monospecific alloantisera against Ag-B haplotypes 1 through 8 with the exception of the Ag-B4 group. This group was shown to include the Ag-B4 (H-1a), Ag-B9 (H-1d), and Ag-B10 (H-1f) haplotypes.

The mixed lymphocyte response (MLR) of peripheral blood lymphocytes from individual wild rats was established by testing against irradiated splenic lymphocytes from inbred rat strains representing seven MLR types. Identity reactions against one, and in many cases two, inbred MLR types were detected in 54 of 59 wild rats successfully tested. Two-way MLR testing established that the identity reactions between wild and inbred animals were reciprocal and that identity reactions could be shown to exist between wild rats from widely separated geographical locations.

The frequency of individual Ag-B(H-1) and MLR phenotypes was clearly different in rats from separate geographical locations. The English population was the most homogenous with only one Ag-B(H-1) and two MLR phenotypes, whereas the Pittsburgh population was more heterogenous and contained seven Ag-B(H-1) and three MLR phenotypes. Although occasional animals could not be characterized for their Ag-B or MLR type, the number of such animals was small, indicating that the degree of genetic polymorphism in the loci controlling these phenotypes in rats is restricted.
Guinea Pig Antiserum to Mouse Cytotoxic T Lymphocytes and Their Precursors
Authors: Rothstein Thomas L. , Mage Michael G. , Mond James , McHugh Louise L.
Published: December 2022

An antiserum specific for cytotoxic T lymphocytes (CTL) and their precursors has been prepared by immunization of strain 13 guinea pigs with a partially purified population of splenic CTL from BALB/c mice immunized with the C57 T cell lymphoma EL4. After absorption with EL4 cells, this antiserum (in the presence of C) totally abolished the activity of in vivo generated CTL and CTL precursors, although it killed only 20% of total Ig- spleen cells. Antibody activity was independent of the H-2 or Ly allotypes of the cells being treated. Helper activity and proliferative responses to concanavalin A and to phytohemagglutinin were unaffected. There was a variable effect on the mixed lymphocyte reaction, and a partial inhibition of graft-vs-host activity.
Functional and Biochemical Properties of the Early Classical Complement System of Mice
Authors: Goldman Margaret B. , Bangalore Shrikar , Goldman John N.
Published: December 2022

Mouse serum and EDTA plasma were subjected to low ionicity precipitation, gel filtration, and ion exchange chromatography in an attempt to purify C1, C4, and C2 to functional and chemical homogeneity. In marked contrast to human and guinea pig components, those of the mouse could not be separated by these techniques. Except for partial separation of C1 from C4 and C2 on DE-52 cellulose columns with EDTA in the eluting buffers, there was no separation of those three components on ion exchange chromatographic columns, Sephadex G-200 gel filtration columns, or with precipitation of euglobulins from serum or plasma. Generation of EAC142 by incubation of EA in whole serum followed first order kinetics when mouse serum was used and second (or greater) order kinetics when human or guinea pig sera were used. Generation of EAC142 by incubation of EA in whole mouse serum followed by incubation in EDTA containing buffers resulted in rapid loss of all three activities from the cell. These experiments indicated that there were significant differences between the early classical C system of mice and those of human and guinea pig. In addition, they indicated that under a variety of in vitro conditions, murine C1, C4, and C2 behaved biochemically and functionally as a unit. The reasons for the major differences in behavior of the murine C components will not become clear until methods to stabilize their function are found so that they can survive multiple purification steps.
Immunochemical Properties of the Murine Ss Protein
Authors: Goldman John N. , Gleason Kristine R. , Goldman Margaret B.
Published: December 2022

Murine Ss protein was found to exist in two forms, distinguishable by electrophoresis, ion exchange chromatography, and gel filtration chromatography. Mouse blood collected directly into EDTA yielded electrophoretically slow Ss protein which eluted from DE-52 cellulose in fractions that also contained C3 and eluted from Sephadex G-200 in fractions containing molecules of 160,000 daltons when high ionicity buffers were used. Mouse serum yielded an electrophoretically faster Ss protein that eluted from DE-52 cellulose well after C3 and eluted from Sephadex G-200 in fractions containing molecules of approximately 128,000 daltons when buffers of high ionicity were used. Incubation of serum led to increasing proportions of the smaller form with time. With appropriate conditions, Ss protein antigen could be separated from C4 functional hemolytic activity on ion exchange chromatography and gel filtration. EAC142 that had been generated by incubation of EA in fresh B10.D2 new serum lost all functional C1, C4, and C2 but not Ss protein from their surface when incubated in EDTA-containing buffers. Treatment of mouse serum with the anti-complementary factor of cobra venom left the C1, C4, and C2 titers unchanged but caused the serum to lose its ability to generate cells that could be agglutinated with anti-Ss antiserum. Similarly treated human serum retained its C1, C4, and C2 activity and its ability to generate cells agglutinable with anti-C4 antiserum. Treatment of mouse serum with NH4OH caused only minimal loss of C4 function but caused the serum to lose its ability to generate cells that could be agglutinated with anti-Ss antiserum. Similarly treated human serum lost both C4 function and the ability to generate cells that could be agglutinated with anti-C4. Thus, mouse Ss protein, an antigenic homologue of human C4, was shown to have several functional and physicochemical differences from human C4. Genetic, evolutionary, and immunologic aspects of this are discussed.
Detection and Partial Characterization of Human T and B Lymphocyte Membrane Antigens with Antisera to HSB and SB Cell Lines
Authors: Anderson Joy K. , Metzgar Richard S.
Published: December 2022

Antisera were produced in rabbits and monkeys by immunization with either HSB (T cell) or SB (B cell) lymphoblastoid cell lines. With a C-dependent microcytotoxicity assay, appropriately absorbed antisera to HSB were strongly reactive with human thymocytes, T lymphoblastoid cell lines, and some acute lymphoblastic leukemia cells (>90% lysis). Approximately 75% of normal peripheral blood lymphocytes (PBL) were lysed by the antisera. Reactivity was at background level when monocytes and cells from patients with chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), and most acute myelocytic leukemia (AML) and acute myelomonocytic leukemia (AMML) were tested against the antisera. Further absorption of anti-HSB sera with CML cells or platelets did not remove serologic reactivity, whereas absorption with thymus, PBL, or concanavalin A- (Con A) stimulated PBL removed reactivity against all cells tested. Antisera produced to SB and absorbed with HSB were strongly reactive with B cell lines, CLL cells, tonsil, adenoid, enriched B cell preparations from normal PBL, and cells from most AML and AMML patients. Additional absorption of anti-SB sera with thymus, CML cells, or platelets did not remove reactivity. Anti-SB sera that were additionally absorbed with CLL cells were negative against all cells tested.

Sodium deoxycholate- (DOC) solubilized membrane preparations from a variety of cells were labeled with 125I and subjected to radioimmunoprecipitation (RIP) and polyacrylamide gel electrophoresis (PAGE) techniques. Unlabeled DOC-solubilized membrane antigens of HSB were able to inhibit the cytotoxic reaction of antisera to HSB with PBL and thymocytes. The reactivity of SB antisera with CLL target cells was not affected by the solubilized HSB antigens. Unlabeled DOC solubilized membrane preparations from HSB, PBL, and thymocytes were able to block the RIP activity of anti-HSB sera.

One major peak of radioactivity (m.w. approximately 150,000 daltons) was consistently observed during PAGE of reduced immunoprecipitates of either monkey or rabbit anti-HSB sera and labeled surface antigens from HSB, PBL, or thymus. Treatment of the immunoprecipitates with additional 2-mercaptoethanol or reduction with dithiothreitol failed to alter the observed m.w. of the antigen. No significant peaks were observed in PAGE when the same antigens were precipitated with either normal sera of antisera to SB.

PAGE of reduced immunoprecipitates of absorbed anti-SB sera and labeled SB membranes revealed two peaks of radioactivity, corresponding to m.w. of 38,000 and 30,000 daltons. When labeled membrane preparations from SB, K562, or CLL cells were precipitated with absorbed antisera to HSB, no significant peaks were observed.