bluuebunny/crossref_metadata_2025 huggingface dataset:
Bursectomy and Thymectomy of Obese Strain (OS) Chickens with Spontaneous Autoimmune Thyroiditis and Simultaneous Experimental Allergic Encephalomyelitis
Authors: Wick Georg , Steiner Renate
Published: December 2022

Experimental allergic encephalomyelitis (EAE) was induced in obese strain (OS) chickens with spontaneous autoimmune thyroiditis (SAT). The effects of surgical bursectomy (BE) in ovo on the 18th day of incubation and of thymectomy (TE) on the day of hatching on the development of these two diseases were studied within the same animal. EAE was elicited in 6- and 11-week-old unoperated, thymectomized or bursectomized OS chickens by a single injection of a saline extract of bovine spinal cord and complete Freund's adjuvant, with Pertussis vaccine as a coadjuvant. Unimmunized birds and chickens injected with bovine liver extract served as controls. Neonatal TE led to an increase in the severity of SAT irrespective of the consecutive immunization, but resulted in a decrease of EAE. BE in ovo entailed a significant decrease of SAT, but also a marked suppression of EAE. Thus the bursa dependency of SAT in OS chickens has been confirmed, while both bursa-derived and thymus-derived lymphoid cells appear to be involved in the development of EAE in this strain.
Macrophage Accumulation, Division, Maturation, and Digestive and Microbicidal Capacities in Tuberculous Lesions
Authors: Dannenberg Arthur M. , Ando Masayuki , Shima Kiyoshi
Published: December 2022

Macrophage turnover and activation were evaluated in dermal BCG lesions produced in normal and tuberculin-positive rabbits. Turnover, i.e., the rate of entry of mononuclear cells (MN) into the lesions and their rate of dealth, was studied by labeling the MN with tritiated thymidine (3HT). Their activation was studied by staining the MN for the lysosomal enzyme β-galactosidase.

A single pulse of 3HT was given 1 day before, or 6, 14 or 27 days after the tuberculous lesions were begun. Five days after the injection, 19 to 33% of the MN in the lesions were labeled with 3HT and their fate could be followed by means of periodic biopsies, each on a different lesion.

3HT-labeled MN entered BCG lesions of reinfection more rapidly than primary lesions. Such MN also died more rapidly in the former. Thus the presence of preexisting tuberculin hypersensitivity markedly accelerated MN turnover.

This conclusion was confirmed in the experiments in which 3HT was given 6, 14 or 27 days after primary BCG lesions were begun. During the week following the 14-day 3HT injection (when tuberculin hypersensitivity was strong), the highest rate of MN entry and the highest rate of MN death were found.

Delayed hypersensitivity to the tuberculin-like products of the bacilli was also correlated with increased numbers of 3HT-labeled, β-galactosidase-positive macrophages. The level of this lysosomal enzyme is a measure of the macrophage's state of activation and reflects its microbicidal capacity, i.e., the degree of its nonspecific cellular immunity. The 3HT-labeling studies just outlined showed that the rate of development of 2- to 4-plus galactosidase activity was most rapid in MN that entered the lesions between 14 and 21 days. At this time the host was highly sensitive to the antigen, and this antigen had probably reached a relatively high concentration. A more rapid development of high β-galactosidase levels also occurred in the MN present in BCG lesions of reinfection.

This work has changed our concept of the pathogenesis of tuberculosis. Formerly, tuberculous lesions were thought to be rather static, and macrophages were thought to become activated and develop antimicrobial immunity gradually. Now such lesions are thought to be dynamic. Macrophages constantly enter and constantly die, and the speed with which they become activated and increase their microbicidal ability seems to be of crucial importance.
A Predictive Equation for the Primary Immune Response of Mice to Adsorbed Tetanus Toxoid As a Function of Dose of Antigen and Dose of Adjuvant
Authors: Levine Leo
Published: December 2022

Of all the adjuvants used in experimental immunology, the insoluble aluminum compounds comprise the only type permitted in human immunization. They confer a 3- to 10-fold higher primary immune response to the usual protein antigens over that available from the plain fluid state, and such priming enhances the secondary response 20- to 40-fold. Immunologists have long been interested in this more complex dose-response relationship with two independent variables, dose of antigen interacting with dose of adjuvant. Applying the data of a human field trial by using four preparations of AIPO4-adsorbed tetanus toxoid with different antigen-adjuvant ratios, the author previously derived an equation that predicted the mean primary serum antitoxin response of subject groups from the dose of antigen and adjuvant injected. The present paper demonstrates the fit of this equation, with the constant terms changed, to the published data of another investigator who used mice to test 14 adsorbed tetanus toxoid preparations differing in antigen-adjuvant ratio. The probit-survival primary responses of the mice were predicted with reasonable accuracy for all 14 preparations by entering the dose of antigen and of adjuvant of each into the equation. The implications for biologic standardization and for the mechanism of the primary immune response are discussed.
Anti-DNA Plaque-Forming Cells (PFC) and Rosette-Forming Cells (RFC) in Spleens of NZB/W F1 Female (B/W) Mice
Authors: Clark C. , Bell D. A. , Vaughan J. H.
Published: December 2022

NZB female × NZW male F1 hybrid (B/W) mice spontaneously develop an autoimmune disease similar to human systemic lupus erythematosus (SLE)4 characterized by the appearance of antinuclear antibodies and fatal glomerulonephritis (1). Although a variety of autoantigens may be involved, anti-DNA has been strongly implicated in the pathogenesis of the renal disease (2, 3). The detectable anti-DNA in the serum is directed predominantly to determinants on single stranded DNA (4, 5).

One of the critical questions with respect to autoimmune diseases in general is whether lymphoid cell function is normal, or if in fact there is altered function of thymic dependent (T) or thymic independent (B) lymphocytes. Although other investigators have approached this problem by examining the cellular response to heterologous antigens in the B/W mouse, similar approaches to autoantigens have not been reported. The present study describes preliminary experiments which demonstrate that the spontaneously occurring immune response to single stranded deoxyribonucleic acid (S-DNA) in B/W mice can be readily detected and quantitated by the hemolytic plaque assay.
The Production of a Monocyte Chemotactic Factor by Agammaglobulinemic Chicken Spleen Cells
Authors: Altman Leonard C. , Kirchner Holger
Published: December 2022

The classification of lymphocytes into bursal derived (B)2 and thymic derived (T) subpopulations has been a significant advance in our understanding of lymphocyte biology. A variety of immune functions have been attributed to each of these subpopulations, while other functions depend on a cooperation between both cell types (1). It has been suggested that lymphokines, the biologically active substances produced in vitro by activated lymphocytes (2, 3) are produced by T lymphocytes (2). However, little definitive experimental evidence to substantiate this conclusion has been reported. The agammaglobulinemic (aγ) chicken has been shown to be largely depleted of B cells (4, 5) and represents an excellent model in which to investigate the contributions of B and T cells to various aspects of the immune response (6). In the present investigation we have adapted a recently described assay of mononuclear leukocyte (MNL) chemotaxis (7) to study lymphokine production by mitogen-stimulated normal and aγ chicken spleen cells.
The Effect of Enzyme Digestion on the Antigenicity and Biologic Activity of Clq
Authors: Wiger Donna , Natvig Jacob B.
Published: December 2022

The human complement component Clq was found to be sensitive to the action of pepsin and papain. Treatment with these two enzymes under certain conditions caused a marked decrease in the ability of Clq to agglutinate latex particles coated with IgG. Gel diffusion studies also showed the presence of components antigenically deficient when compared to non-treated Clq. After ultracentrifugation of the pepsin-treated Clq the antigenically deficient components were located in fractions having much lower sedimentation rates than native Clq.

Reduction and alkylation of Clq also resulted in decreased ability to agglutinate the latex particles and caused the formation of antigenically deficient components. However, the components with lower sedimentation constants than Clq were detected only if conditions causing decreased non-covalent interaction were utilized.
The Immune Response in Mice to the Haptenic Determinant Dnp Coupled to a Thymus-Independent Carrier (Levan)
Authors: Guercio P. Del , Leuchars Elizabeth
Published: December 2022

A thymus-independent antigen (levan) can function as a carrier and induce the synthesis of antibodies to DNP. The DNP-levan conjugate is able to elicit only 19S antibody against the hapten. This immune response is not affected in thymectomized mice, but is abolished in mice tolerant to levan. A mechanism involving cooperation between hapten-specific B lymphocytes and carrier-specific B lymphocytes is suggested to account for these findings.
Measurement of Antibody Release from Single Cells. II
Authors: Hiramoto Raymond N. , Hamlin N. Marlene , McGhee Jerry R.
Published: December 2022

A method for determining the relative amounts of antibody released from single cells in a population is presented. The procedure is based on estimating the minimal amount of antibody molecules required to give complete lysis in a given plaque volume. The results show that the average cellular rate from five populations studied was 360 molecules/cell/sec. The value is regarded as a minimum estimate of the average rate.
The Release of Granule Enzymes from Human Neutrophils Stimulated by Aggregated Immunoglobulins of Different Classes and Subclasses
Authors: Henson P. M. , Johnson H. B. , Spiegelberg H. L.
Published: December 2022

Aggregated human myeloma proteins of subclasses IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2 and normal IgG have been shown to react with human neutrophils in a serum-free medium and to induce release of the lysosomal enzyme β glucuronidase to the exterior of the cells. In contrast, IgD, IgE and IgM macroglobulins did not stimulate this release. Incubation of neutrophils with insoluble IgG and IgA aggregates resulted in adherence of the aggregates to the neutrophils and their phagocytosis. During this process, release of granule enzymes, but not of the cytoplasmic enzyme lactic dehydrogenase, occurred, indicating a selective process rather than one of cell lysis. Soluble aggregates did not induce liberation of enzymes when incubated with the neutrophils in suspension, but when bound to micropore filters to serve as nonphagocytosable surfaces, they stimulated adherent cells to release their granule contents. This latter process has previously been shown to result from direct extrusion of neutrophil granules to the exterior.
A Simple Efficient Procedure for the Preparation of Fragments Corresponding to the Variable-Half and Constant-Half Portions of Bence Jones Protein
Authors: Seon B.-K. , Roholt O. A. , Pressman D.
Published: December 2022

A simple efficient procedure was developed for the preparation of two fragments corresponding closely to the variable-half (VL) and the constant-half (CL) portions of Bence Jones protein. The fragments resulted from controlled peptic digestion of a type K Bence Jones protein. The close correspondence of the VL and CL fragments to the variable half and the constant half of the Bence Jones protein, respectively, was shown by immunoelectrophoretic analyses and a study of the amino acid compositions of the isolated fragments and indicates that pepsin cleaves Bence Jones protein close to the switch point between the variable half and the constant half. Our results thus support the domain hypothesis. The procedure is based upon our finding that peptic digestion of Bence Jones proteins at 37°C yields only the VL fragment and some remaining intact Bence Jones protein whereas at 55°C, only the CL fragment results. Intact type K Bence Jones protein (no prior reduction-alkylation) was digested with pepsin at 37°C for the preparation of the VL fragment, and at 55°C for the CL fragment. The fragments were isolated from the respective digests by using in each case a single gel filtration. The yields were markedly high, being 70% for the VL fragment and more than 60% for the CL fragment.
Cytotoxic Activity of Lymphocytes
Authors: Walker Sharyn M. , Lucas Zoltan J.
Published: December 2022

Characteristics of the cytotoxicity caused by supernatants of lymphocytes activated in culture are presented. The cytotoxic factor (lymphotoxin, LT) from human spleen cells incubated with phytohemagglutinin in serum-free medium was partially purified by gel electrophoresis. The cells left on the monolayer after incubation with LT demonstrated no detectable abnormalities. Increased permeability of the cell membrane to 86Rb did not occur. Incorporation of 3H-leucine or 3H-thymidine was unimpaired when related to the number of viable residual cells. Cells died and fell off the monolayer even as 3H-thymidine incorporation per cell was unchanged. Kinetic analysis indicates that cell death increases exponentially with time and is second order for the concentration of LT. Cell death is caused by a mechanism other than a general inhibition of protein or DNA synthesis.
The Rise and Fall of Antibody Avidity at the Level of Single Immunocytes
Authors: Doria G. , Schiaffini G. , Garavini M. , Mancini C.
Published: December 2022

Antibody avidity for trinitrophenyl (TNP) was investigated at the level of single immunocytes in spleen cell populations from mice immunized by intravenous injection of TNP horse red blood cells. Early variations of avidity were observed before any significant increase in anti-TNP plaque forming cells (PFC) number was noticed. Subsequently, avidity increased with time, reached a maximum when the number of PFC attained the peak value, and then decreased as PFC were disappearing from the spleen. A 104-fold variation in antigen dose did not affect this pattern. Avidity and PFC number decreased with lower antigen doses.
Characteristics of Complement Fixation by Aggregated IgE
Authors: Ishizaka Teruko , Soto Carmen S. , Ishizaka Kimishige
Published: December 2022

Aggregated IgE fixed late components of complement (C), i.e., C3–C9, through an alternate pathway, bypassing C1, 4, and 2 whereas monomeric IgE did not. The aggregate, however, did not fix purified C3, suggesting that C3 activator system is involved in the reaction. On a weight basis, aggregated Fc had higher C-fixing activity than aggregated IgE, and aggregated F(ab′)2 was less active. The results indicated that structures essential for C fixation are present in the Fc portion of IgE molecules. A mild reduction and alkylation of IgE, which degraded structures involved in passive sensitization of monkey skin, did not affect the C-fixing properties of the protein.
The Fractionation of Immunoglobulins with Insolubilized Concanavalin A
Authors: Weinstein Yacob , Givol David , Strausbauch Paul H.
Published: December 2022

A method is described for the facile separation of IgG and IgM based on their differences in carbohydrate content. When a preparation of specific anti-hapten antibody is applied to a column of Sepharose-insolubilized concanavalin A, an unbound fraction containing IgG is washed through the column uncontaminated by other classes of immunoglobulins. The column-bound protein is then eluted with 0.2 M α-methylmannoside. This material contains all the IgM plus about 5% of the original IgG. This concanavalin A-bound IgG appears to be absorbed specifically to the column since reapplication of this material results in the majority of it being rebound, due perhaps to some structural variation in the carbohydrate moieties it contains.
The Qualitative Receptor Analysis
Authors: Felix A. , Olitzki L.
Published: December 2022

In an elaborate clinical investigation made upon typhoid patients Felix (1) was able to report on the results of qualitative receptor analysis, that from a practical point have developed new points of view with reference to certain problems of the serology of typhoid fever.
On Anaphylaxis with Heated Serum
Authors: Furth J.
Published: December 2022

It is usually assumed that the phenomenon of anaphylaxis depends on the presence of soluble protein and that such proteins as horse serum, when made insoluble by heat, are deprived of their anaphylactic toxicity. Nevertheless, there is little doubt that protein heated to the boiling point acts as antigen in precipitation as well as in complement fixation reactions and produces local inflammation in animals sensitized with it (1). These observations which indicate that anaphylaxis and precipitation are independent phenomena (Pick and Yamanouchi (2)) suggested a study of anaphylaxis produced by heated serum. The heated antigen has been used for sensitization, since in earlier studies due attention has not been paid to the possibility that heated serum may effectively sensitize an animal to heated antigen.

The investigations of Besredka (3), Rosenau and Anderson (4), Doerr and Russ (5), Kraus and Volk (6), Arthus (7), Uhlenhuth and Haendel (8) have shown that the sensitizing property of serum resists heat while its toxicity, it appears, is readily destroyed when it is heated to about 90°C.
On the Rôle of the Reticulo Endothelial System in the Tuberculin Hypersensitiveness
Authors: Freund J.
Published: December 2022

Splenectomy and injections of iron and trypan blue hinder the development of the hypersensitiveness in tuberculous guinea pigs.

Splenectomy in itself has no effect upon the development of hypersensitiveness.

Injections of iron sugar and trypan blue in the amounts used do not lessen the already developed skin sensitiveness.

These observations and a general consideration indicate that the reticulo-endothelial system has a function in the mechanism of the development of the tuberculin hypersensitiveness, which creates an analogy between the development of the tuberculin hypersensitiveness and the production of antibodies.
Heterophil Antigen-Antibody Reactions in Relation to the Serum Diagnosis of Syphilis by Precipitation
Authors: Hooker Sanford Burton
Published: December 2022

While studying the reactivity of the serum of a rabbit which had been injected with hen erythrocytes, and which had responded with an abundant production of heterophil (antisheep) hemolysin (Kritchewsky, Hyde), its precipitating effect on several Wassermann and Kahn “antigens” was tested. The serum flocculated powerfully with the lipoids extracted from horse heart according to Kolmer's method, and those contained in a plain alcoholic extract of guinea pig heart but it had no such effect when mixed with Kolmer's beef heart antigen nor with beef, human, and swine heart extracts prepared by Kahn's technic. An immune guinea pig antihen-cell serum lacked this specific precipitating property—its lysin being isophil.

Observation of the specific reactions of this hemolytic serum with heterophil “lipoids” (horse and guinea pig) led to the thought that when such extracts are employed in diagnostic precipitation (or fixation) tests there would often be present conditions which might result in falsely positive reactions.
Synergism between Thymocytes and Lymph Node Cells in the Graft-Versus-Host Response. Effect of Cortisone Treatment of the Thymus Cell Donor
Authors: Blomgren Henric
Published: December 2022

Combinations of thymic cells from 1-month-old and lymph node cells from 6-month-old C57BL mice were tested for their capacity to give a synergistic effect in the graft-versus-host reaction (G-v-H) when injected into young A/Sn × C57BL F1-hybrids. No clear synergism was observed between normal thymic cells and lymph node cells.

Two days after cortisone treatment of the thymus cell donors the cell number in the thymus had dropped to 5% of its original value and the relative G-v-H reactivity exhibited a 15-fold increase. There was no evidence, however, of an enrichment of cells capable of amplifying the G-v-H reaction. On day 12 after steroid treatment the G-v-H reactivity of the organ had decreased, but there was a marked tendency of the cell population to act synergistically with lymph node cells. After another 8 days the thymus cell population had regained its G-v-H reactivity and diminished its “amplifying” capacity. The transformation of amplifying cells into G-v-H reactive cells is a possible explanation of this phenomenon.
Rabbit Immunoglobulin Light Chains: Correlation of Variable Region Sequences with Allotypic Markers
Authors: Waterfield M. D. , Morris J. E. , Hood L. E. , Todd C. W.
Published: December 2022

Evidence that the group b allotype markers correlate with certain mixtures of variable region sequences in rabbit immunoglobulin light chains is presented. The amino terminal sequences of κ chains from heterogeneous rabbit immunoglobulin of differing allotypes were examined by using an automatic protein sequenator. Each of the four allotypes has a distinctive pattern of amino acid residue alternatives over the first 20 amino terminal residues. Since previous studies have shown that amino acid sequences correlating with the allotypes are present in the constant region, our observations suggest that constant and variable region genes of κ chains may be closely linked.
Granulocyte Chemotaxis: An Improved in Vitro Assay Employing 51Cr-Labeled Granulocytes
Authors: Gallin John I. , Clark Robert A. , Kimball Harry R.
Published: December 2022

A radioassay of granulocyte chemotaxis employing chromium-51 labeled cells and a double micropore filter system is presented. In comparison to the conventional Boyden chamber technique, this method offers significant advantages including: a) elimination of tedious filter staining and microscope counting resulting in a considerable saving in laboratory time, b) elimination of observer subjectivity, c) excellent sensitivity over a wide range of cell responses with no increase in variability, d) ability to monitor cellular events in all compartments of the chemotaxis chamber and e) a means for standardization of neutrophil chemotaxis among different laboratories.
Differential Effects of Cyclophosphamide on the B and T Cell Compartments of Adult Mice
Authors: Stockman Gail D. , Heim Lyle R. , South Mary Ann , Trentin John J.
Published: December 2022

Adult mice were given a single sublethal dose of cyclophosphamide. Effects of the drug on the bone marrow-derived (B) and thymus-derived (T) cell compartments were measured by studies on lymphocyte transformation, lymphoid tissue morphology and responsiveness to a primary injection of keyhole limpet hemocyanin. These studies indicated that the effect of cyclophosphamide on the B cell compartment was more severe and longer-lasting than the effect on the T cell compartment.
Enhancement of Immunologically Induced Granule Exocytosis from Neutrophils by Cytochalasin B
Authors: Henson Peter M. , Oades Zenaida G.
Published: December 2022

Polymorphonuclear neutrophil leukocytes (neutrophils) release granule enzymes to the extracellular medium after adherence to immune complexes or aggregates attached to surfaces too large to be phagocytosed (1–4). A number of facts have emerged in the past few years that define this phenomenon. The release has been shown to occur by direct exocytosis of granules along the neutrophil membrane in contact with the surface but not along the free, unattached membrane of the cell (2, 5). It requires stimulation by aggregated immunoglobulin G immune complexes, or complement on the surface, but not aggregated IgM, IgD, IgE or proteins unrelated to immunoglobulins (6). The release of granular enzymes requires calcium in the external medium and glycolytic metabolism on the part of the neutrophil. It is accompanied by stimulation of glucose oxidation by the hexose monophosphate pathway, but this is not required for the secretion. The release can be inhibited by diisopropyl fluorophosphate and agents which increase intracellular cyclic AMP (7, 8).
Adjuvanticity of Mycobacterial RNA and Poly A:U for Induction of Experimental Allergic Encephalomyelitis in Guinea Pigs
Authors: Gumbiner Carl , Paterson Philip Y. , Youmans Guy P. , Youmans Anne S.
Published: December 2022

Youmans and Youmans (1, 2) have reported that mycobacterial ribosomal or RNA preparations rival viable attenuated mycobacteria (H37Ra) in their capacity to engender cellular immunity to challenge with virulent mycobacteria (H37Rv). More recently, these workers have discovered mycobacterial RNA preparations to augment humoral as well as cellular immune responses (3). Concurrent studies by a number of investigators (4–7) have demonstrated synthetic polynucleotides to potentiate immune responses to a variety of antigenic stimuli. These and other studies have served to focus attention on the important role of microbial and synthetic double stranded RNA as immunobiologic adjuvants.

Mycobacteria are a prerequisite adjuvant component for sensitization of guinea pigs to neuroantigens and regular induction of experimental allergic encephalomyelitis (EAE)3 in this animal species (8). While EAE-induction has been reported with mycobacterial lipid or lipopolysaccharide fractions and WaxD, as well as other microorganisms, none has equalled the biologic potency of killed whole mycobacteria for production of this experimental autoimmune disease (9–12).
Immunobiology of the Autoantibody Response
Authors: Linder Ewert , Edgington Thomas S.
Published: December 2022

Antierythrocyte autoantibodies directed against HB autoantigen, a cryptic site on the murine erythrocyte membrane, have been implicated previously in both direct cellular injury and autologous immune complex injury in NZB mice. Identification, isolation and characterization of this autoantigen in soluble form are sought in order to facilitate studies of the immunobiology and cellular immunology of the autoimmune response. Anti-HB autoantibodies are qualitatively neutralized not only by soluble autoantigen present in mouse plasma but also by the purified low density lipoprotein (LDL) fraction of plasma or serum. The LDL-associated HB autoantigen fraction, which is immunochemically analogous to the membrane bound autoantigen HB, appears to represent the soluble erythrocyte autoantigen SEA-HB. This soluble erythrocyte autoantigen is apparently not identical to the LDL lipid or apoprotein per se, since anti-LDL antibody does not react with erythrocyte surface HB determinants and a generic parallel but quantitative discrepancy is observed between the concentration of LDL and SEA-HB in immunoabsorption experiments. These data suggest that SEA-HB may be carried by plasma lipoproteins due to lipophilic or other properties of this autoantigen or that it may represent a minor subpopulation of murine LDL molecules.
Studies on the Mechanism of Lymphocyte-Mediated Cytolysis
Authors: Henney Christopher S. , Bubbers J. Eric
Published: December 2022

The role of divalent cations in the thymus-derived (T) lymphocyte-mediated cytolysis of DBA/2 mastocytoma cells by immune allogeneic (C57BL) splenic lymphocytes has been studied. In the presence of 0.001 M EDTA no specific cytolysis (51Cr release) was demonstrable. Cytolysis was restored upon addition of either an excess of Ca2+ or a much larger excess of Mg2+. When EDTA was added to an ongoing cytolytic event there was a considerable lag period of the order of 1 hr before inhibition was demonstrable, indicating the existence of cation dependent and independent phases of cytolysis. Although cytolysis did not occur when lymphocytes and target cells were incubated together in the initial presence of EDTA, upon readdition of cations the rate of cytolysis was increased significantly from that observed when the interacting cells were not pre-incubated with inhibitor. These findings suggest that initial steps along the cytolytic pathway proceed in the absence of cations, but fall short of cytolysis.

When specific target cell lysis was inhibited by cytochalasin B or by prostaglandin E2 (PGE2), the substitution of EDTA for these inhibitors further suppressed cytolysis. On the other hand, cytolysis proceeded in an unimpeded manner when EDTA-inhibited reaction mixtures were washed and confronted with the same concentrations of cytochalasin B or PGE2 which completely inhibited de novo cytolysis.

These findings demonstrate that the cytolytic event proceeds in discrete stages. The events which are inhibited by cytochalasin B or the prostaglandins are clearly distinct from, and occur before, those stages which require divalent cations and are inhibited by EDTA.
Monoclonal IgA and IgM in the Serum of a Single Patient (SC)
Authors: Silverman Andrew Y. , Yagi Yasuo , Pressman David , Ellison Rose R. , Tormey Douglas C.
Published: December 2022

“Monoclonal” IgA(κ) and IgM(κ) present in the serum of a patient (SC) were previously shown to have identical κ light chains and great similarity in the variable portions of their heavy chains. Immunofluorescence technique was used in this study on bone marrow smears of SC to identify the cells producing these proteins. IgA and IgM were found to be produced in separate cells. No cells containing both proteins were detected. These results strongly suggest that the clones of the cells producing SC IgA or SC IgM were derived from a single stem cell and support the view that the variable and constant portions of immunoglobulin chains are regulated by two different genes.
Syngeneic Mixed Lymphocyte Reaction between Thymocytes and Peripheral Lymphoid Cells in Mice: Strain Specificity and Nature of the Target Cell
Authors: von Boehmer H. , Adams P. B.
Published: December 2022

The syngeneic mixed lymphocyte reaction between thymus cells and peripheral lymphoid cells was investigated in CBA, C57BL and BALB/c mouse strains. Four-day-old thymus cells from CBA and BALB/c mice proliferate in culture with syngeneic Mitomycin C-treated lymphoid cells. The reaction is smaller (or not significant) when the thymus cells are prepared from older mice. In contrast, 4-day-old C57 thymus cells do not respond significantly to syngeneic lymphoid cells, although they respond well to allogeneic cells. CBA and BALB/c peripheral lymphoid cells may possess an antigen “other than H2” responsible for the stimulation of syngeneic thymus cells.

The target cell for the syngeneic mixed lymphocyte reaction in CBA mice is a lymphocyte present in spleen, peripheral blood, lymph nodes and peritoneal exudate.
Blocking of the Hemolytic Expression of the Classical Complement Sequence by Products of Complement Activation via the Alternate Pathway
Authors: Koethe Susan M. , Austen K. Frank , Gigli Irma
Published: December 2022

Activation of the complement sequence through the alternate pathway with zymosan or the low molecular weight cobra venom factor (CoVF) generates products which block the lysis of hemolytic cellular intermediates of the classical complement sequence. The euglobulin fraction of the eluate of zymosan and products in CoVF-treated serum bind to EAC14limoxy2 and EAC14limoxy23 to prevent effective hemolytic interaction with C3 and C5, respectively. Analyses of fractions from gel filtration of both the zymosan eluate and CoVF-treated serum revealed blocking in the IgM region of EAC14limoxy2, EAC14limoxy23, and EACl-7 while materials eluting in the trough between the IgM and IgG peaks blocked only EAC14limoxy2 and EAC14limoxy23. Radiolabeled C5 in CoVF-treated serum was shown to be present in the material eluting in both regions representing a shift from the usual position of C5. Only the highest molecular weight material destroyed C9. It is proposed that the blocking of the cellular intermediates is due to complexes ranging in composition from C5 and C6 to C5, C6, C7 and C8, which are hemolytically inactive while maintaining certain functions, such as binding to the appropriate cellular intermediate or to one or more later components in the complement reaction sequence.
Cell Surface Associated Gamma Globulin in Lymphocytes
Authors: Orr K. B. , Paraskevas F.
Published: December 2022

Serum collected from mice 6 hr after immunization with soluble antigens in Freund's complete adjuvant was able to induce various changes of the bone marrow-derived (B)-cell (γ globulin-carrying) and thymus-derived (T)-cell (θ-carrying) populations when incubated with spleen cells from unimmunized mice. First there was an overall increase (by 25%) of all the γ globulin-carrying cells. This increase could be accounted for by an identical increase of the cells carrying γG (7S γ2a) globulin on their surface. At the same time the cell population normally carrying γF (7S γ1) globulin decreased. These changes were detected by the reverse immune cytoadherence technique (RICA) with highly specific hybrid antibodies which detect one class of mouse γ globulins. Second, the treated spleen cells show a decrease of the T-cell population as detected by an anti-ϑ serum. This decrease was of a similar magnitude as the increase of the γG-carrying cells and involved about 40% of the ϑ-carrying cells. Thus the serum reproduces in vitro all the phenomena previously described as taking place in the spleen 6 hr after immunization. When the 6-hr serum from mice injected with radioactively labeled bovine serum albumin was fractionated on a Sephadex G-200 column, all the activities described above were reproduced by a fraction containing 7S γ globulin and antigen. The presence in this fraction of a complex of the antigen with mouse immunoglobulin was confirmed by autoradiography. It is quite possible that the alterations in the populations of B- and T-cells described are due to the interactions of such a complex with these cells in the spleen cell suspension.
Pseudoimmune Precipitation by the Isolated Byssinosis “Antigen”
Authors: Edwards John H. , Jones Brian M.
Published: December 2022

The so-called cotton antigen, a polymer of 5,7,3′,4′ tetrahydroxyflavan 3–4 diol (THF) extracted from cotton plant bracts, precipitated 58% IgG, 54% IgM and 15% IgA in a sample of human serum and concurrently antibody activity in this serum to three physicochemically distinct antigens was reduced by 50%, 52% and 62%, respectively. THF also reacted with five myeloma IgG at dilutions equal to, or higher than, comparable dilutions of human serum and gave positive gel diffusion reactions with H and L chains, Fab and Fc pieces of IgG.

On the basis of these results it is suggested that human immunoglobulins are nonspecifically precipitated by the tannin-like polymer.
Natural Antiglobulin Antibodies in Primate Sera
Authors: Baxter N. H. , Malley A. , Mandy W. J.
Published: December 2022

Papain and pepsin agglutinators showing specificity for autologous or homologous Fab and (Fab′)2 have been demonstrated in normal human and rabbit sera. In this study papain and pepsin agglutinators were shown to be present in normal rhesus serum. The evidence also suggests that these agglutinators are present in members of other families of the order Primates.

In a comparative study, individual sera from several members of Primates representing the families Hominidae, Pongidae, Cercopithecidae, Cebidae and the Prosimians were tested for agglutinator activity against the Fab and (Fab′)2 fragments of human and rhesus IgG. The pepsin agglutinators in each of the families studies reacted with rhesus (Fab′)2 fragments, but only members of the families Hominidae, Pongidae and Cercopithecidae reacted with human (Fab′)2 fragments. Human papain agglutinators were specific only for human Fab fragments. Likewise Cercopithecoid papain agglutinator reacted only with rhesus Fab fragments.
Prevention of Marrow Allograft Rejection with Radioactive Strontium: Evidence for Marrow-Dependent Effector Cells
Authors: Bennett Michael
Published: December 2022

Mice injected with the bone-seeking isotope, 89Sr, were depleted of marrow erythropoietic progenitor cells but maintained normal or near-normal numbers of splenic erythropoietic progenitor cells, lymphoid tissue alloantigen-sensitive units responsible for graft-vs-host reactions, and splenic antigen-sensitive units responsive to sheep erythrocytes which give rise to antibody-producing cells. The ability of such mice to reject marrow allografts was suppressed. It is concluded that a separate class of marrow-dependent immunocompetent cells exist, now called “M cells,” which are responsible for rejection of allogeneic dispersed hemopoietic cells.
Suppression and Selection of B Cells during Lysozyme Tolerance in the Mouse
Authors: Scibienski R. , Sercarz E.
Published: December 2022

Soon after tolerance induction, cross-reactive avian lysozymes terminate tolerance to hen lysozyme (HEL).3 The production of cross-reactive antibody at this stage indicates that the bone marrow-derived (B) cell compartment is still non-tolerant. Later, B cell tolerance is also induced, but it requires a longer exposure and a higher dose of antigen than does induction of thymus-derived (T) cell tolerance.

At a time during recovery from neonatal or adult tolerance to HEL, when HEL-reactive B cells can again be demonstrated, mice challenged with turkey lysozyme (TEL) produce only TEL-specific antibody. The possibility is suggested that TEL-specific B cells compete more favorably for available antigen than do the newly returning, cross-reactive B cells.

The reduced response to challenge in tolerant animals is discussed in relation to a model in which there are stringent stereospecific rules governing the presentation of antigen to B cells.
Fibrosarcoma Induction in Immunosuppressed Mice
Authors: Sanford Barbara H. , Soo Sophie F.
Published: December 2022

Results of an unpublished pilot study performed in this laboratory suggested increased incidence of fibrosarcomas at the site of subcutaneous plastic film implants in immunosuppressed mice as compared with normal mice. Plastic-induced tumors, like most spontaneous tumors, have low antigenicity (1) and should provide a better model than highly antigenic chemically or virally induced tumors for studying the relationship between the immune system and tumor development. The experiment was expanded as is described below.

Ninety C3H/St male mice were operated on at 60 ± 7 days; half of these were thymectomized and the rest sham-thymectomized. Eight days later, whole body irradiation at 350 R was carried out, a procedure previously shown by Miller to produce moderate, prolonged immunosuppression (2). (Concurrent experiments in our laboratory showed significant immunosuppression induced by this method, with 100% of 30 thymectomized, irradiated C3H mice succumbing within 30 days to an inoculum of allogeneic tumor (TA3) which was lethal for only 53 ± 9% of control C3H mice.)
The Physicochemical Characteristics and Purification of Slow-Reacting Substance of Anaphylaxis
Authors: Orange Robert P. , Murphy Robert C. , Karnovsky Manfred L. , Austen K. Frank
Published: December 2022

A preparative high yield procedure for the purification of slow-reacting substance of anaphylaxis (SRS-A) was developed by gas chromatography, gas chromatography-mass spectrometry and high resolution mass spectrometry to determine the required steps and evaluate the final product. The purification procedure was based on consideration of: the stability of the ethanol extracted starting material; the efficiency of desalting by non-ionic chromatography; the removal of phospholipids by base hydrolysis; the extraction of contaminating hydrocarbons, steroids and lipids by silicic acid chromatography and the elimination of other contaminants on the basis of molecular size with Sephadex LH-20 gel filtration. The final product when analyzed by ultraviolet and infrared spectroscopy and high resolution mass spectrometry for the first time demonstrated significant differences between the active preparations and their controls. On a weight basis, this highly purified, acidic, low molecular weight chemical mediator was active at the nanogram level or less.
A Human Mononuclear Leukocyte Chemotactic Factor: Characterization, Specificity and Kinetics of Production by Homologous Leukocytes
Authors: Altman Leonard C. , Snyderman Ralph , Oppenheim Joost J. , Mergenhagen Stephan E.
Published: December 2022

Leukocytes from peripheral human blood when stimulated in vitro with a specific antigen or a nonspecific mitogen produced a soluble factor which was chemotactic for homologous monocytes. The production of this lymphokine proved to be a sensitive correlate of delayed hypersensitivity. A temporal dissociation between both PHA and PPD-induced lymphocyte proliferation and monocyte chemotactic factor production was clearly established. This factor in non-plasma supplemented cultures was present after 20 to 24 hr of incubation; with minimal amounts of added plasma, a shift to earlier production was noted. Glass bead column purified lymphocytes, stimulated with PHA, produced significant amounts of monocyte chemotactic activity. The chemotactic factor was heat stable, nondialyzable, remained active if lyophilized, and had a molecular weight of approximately 12,500. In addition, this material was antigenically distinct from human C3 and C5, precursors of two chemotactic polypeptides derived from complement.
Immunoglobulin Synthesis in Bullfrog Larvae
Authors: Moticka Edward J. , Brown Bruce A. , Cooper Edwin L.
Published: December 2022

The antibody-synthesizing capacity of tadpoles (Rana catesbeiana) has been studied with sheep erythrocytes as antigen. Tadpoles can mount an immune response similar to that seen in higher vertebrates except for a slower kinetic profile. They could not, however, be stimulated to elicit a true anamnestic response. The antibody had characteristics indicating that it probably was all IgM like. Use of the hemolytic plaque technique with various lymphoid organs detected synthesis of antibody in spleen, lymph gland and thymus.
An Antigenic Marker for Human Thymic Lymphocytes
Authors: Smith R. W. , Terry W. D. , Buell D. N. , Sell K. W.
Published: December 2022

Murine thymocytes and thymus-derived lymphocytes possess antigens not normally represented on most other tissues. Antigens of the TL (1), Ly (2), and θ (3) systems can be detected by isoimmunizations with mouse strains differing at the appropriate genetic locus. By immunization of rabbits with mouse lymphoid cells, followed by appropriate absorption procedures, reagents have been produced which can also identify and selectively lyse several different lymphocyte populations (4–6). The availability of immunologic reagents for detecting antigenic differences has greatly facilitated investigations of functional differences among subpopulations of mouse lymphocytes. Similar reagents have not been available previously for the study of human lymphocyte subpopulations. We have investigated the usefulness of absorbed heterologous antihuman thymocyte serum for the detection of antigenic heterogeneity in human lymphoid cells.

Thymus cells were obtained from fetuses and from infants under 1 year of age. Rabbits were injected in the foot pads with 109 cells emulsified with complete Freund's adjuvant.
The Presence of J-Chains in the Intracellular Fluid of IgM-Secreting Murine Tumor Cells
Authors: Raam Shanthi V. , Inman F. P.
Published: December 2022

The cells from the murine tumor HPC 76 secrete IgM. The intracellular fluid was analyzed for the presence of the J-chain. Subsequent to analytical polyacrylamide gel electrophoresis a band was observed which had migrated most rapidly anodally. Its position was similar to that of the ostensible J-chain derived from sulfonated IgM, sulfonated IgA, and the intracellular fluid from the IgA-producing MOPC 315 tumor cells. Intracellular J-chains also were precipitable with specific antiserum. The antibodies recognized identical J-chain antigens in sulfonated IgM, sulfonated IgA, and in the intracellular and extracellular fluids of the HPC 76 cells.
Macrophage migration inhibition in response to experimental mycoplasma pneumoniae infection in the hamster
Authors: Biberfeld Gunnel
Published: December 2022

The migration of peritoneal exudate cells collected from hamsters experimentally infected with M. pneumoniae was inhibited in the presence of M. pneumoniae antigen. Migration inhibition was demonstrable 1 week after intranasal inoculation of the hamsters with M. pneumoniae organisms and reached a peak 2 to 3 weeks after inoculation. The migration inhibition test became positive before specific CF antibodies had appeared.
Fetal Lymphoid Tissues: Antibody Production in Vitro
Authors: Tyan Marvin L. , Ness Dolly B. , Gibbs Priscilla R.
Published: December 2022

It is now thought that in the mature mammal, cell-mediated immune reactions are controlled by a bone marrow-derived, thymuspassaged cell(s) (1–3), while normal antibody production is dependent upon the interactions of macrophages, bone marrow-derived antibody-producing cells and a bone marrowderived, thymus-passaged cell whose specific function(s) is still to some degree in debate (4, 5). Although evidence has been presented that antibody production and specificity in response to “thymus-dependent” antigens results from the reactions between antigen, the antibody-producing cell, and thymus-passaged cells (6), other observations have suggested that limited antibody production and to a great degree the specificity of the antibody, are innately determined by the antibody-producing cells and their precursors (7). Lymphoid precursors, taken from mouse embryos as early as 3 days before the appearance of the thymic rudiment (i.e., on the 9th day of gestation), are able to produce immunoglobulins in thymectomized-irradiated hosts as well as they do in shamthymectomized controls (8); moreover, these cells are capable of a limited antibody response in the absence of the thymus when stimulated with sheep red blood cells (SRBC)2 (7).
Human Leukocyte Migration Inhibition in Agarose Using Four Antigens: Correlation with Skin Reactivity
Authors: Astor Stephen H. , Spitler Lynn E. , Frick Oscar L. , Hugh Fudenberg H.
Published: December 2022

A technique is described for the in vitro evaluation of human cellular immunity by determining the inhibition of the migration of peripheral buffy coat leukocytes in agarose plates after incubation of the cells with and without antigens. The technique is simple to perform and results can be obtained in 18 hr. With three antigens (Candida, PPD and SK-SD), the results correlated well with skin reactivity in normal subjects.
Localization of Endogenous C-Type Virus in the Glomerular Basement Membrane of Aged AKR Mice
Authors: Batzing Barry L. , Hanna M. G.
Published: December 2022

Naturally occurring glomerulonephritis in conventional NZB (1), AKR (2), and RF (3) mice has been associated with an immune complex etiology. In all three cases, it has been demonstrated that the glomerular lesion is a result, in part, of chronic deposition of immune complexes of endogenous Gross virus-specific cell surface and/or soluble antigens with “G natural” antibody. Glomerular lesions of this type also have been observed in AKR (4) and BALB/c (5) mice with experimentally induced leukemias, and in the latter study they were specifically associated with immune complexes involving antigens of the exogenous murine leukemia virus (MuLV).3 At the ultrastructural level, this glomerulonephritis is characterized by marked thickening of localized regions of the glomerular basement membrane (5, 6), which is thought to result from the immune complex deposition (7). We report here the localization of intact endogenous C-type virus within the basement membrane of aged AKR mice showing evidence of chronic glomerulonephritis.