bluuebunny/crossref_metadata_2025 huggingface dataset:
Biologic Significance of Disulfide Bonds in Human IgE Molecules
Authors: Takatsu Kiyoshi , Ishizaka Teruko , Ishizaka Kimishige
Published: December 2022

E myeloma protein, PS, was reduced in different concentrations of dithiothreitol (DTT) for 1 hr followed by alkylation with 14C-iodoacetamide. The affinity of the reduced-alkylated molecules for target cells was evaluated by their ability 1) to sensitize primate skin in a reversed P-K reaction, 2) to sensitize human basophils in a reversed-type histamine release and 3) to block passive sensitization with reaginic antibody. Antibody-ε0 antibody was employed for reversed type reactions to avoid participation of cellbound normal IgE in the reactions. The sensitizing activity of IgE did not change following reduction in 1 mM DTT, which split inter-heavy-light chain disulfide bond. The activity of IgE significantly diminished after reduction in 2 mM DTT followed by alkylation. This treatment resulted in the cleavage of two intra-ε-chain disulfide bonds, which are present between the hinge and the Fd portion of the molecules. The reduced-alkylated protein was capable of sensitizing primate skin and human basophils, however, a much higher concentration of the reduced-alkylated protein than the native protein was required for passive sensitization. The optimal sensitization period for the reversed P-K reaction was 3 hr with the reduced-alkylated protein. The protein had the ability to block passive sensitization with reaginic antibody. The reduced-alkylated protein and the native protein were labeled with 125I, and binding of these proteins with human basophils was examined by autoradiography. The results showed that affinity of the reduced-alkylated protein for basophils was less than that of native protein. Since the disulfide bonds split by 2 mM DTT were not included in the Fc portion of the molecule, the Fc fragment was obtained from the reduced-alkylated protein and was tested for affinity for basophils. It was found that the Fc fragment had higher affinity than the reduced-alkylated protein. Recovery of the affinity by papain digestion strongly suggested that cleavage of disulfide bonds in the Fab portion of the molecules induced conformational changes in the Fc portion which is involved in binding to the target cells. Reduction of IgE with 10 mM DTT followed by alkylation resulted in cleavage of 5 disulfide bonds, which is accompanied by a loss of both sensitizing and blocking activities. The fifth disulfide bond which was cleaved by 10 mM DTT, but not by 2 mM DTT, appears to be an inter-heavy chain disulfide bond in the Fc portion of the ε-chains. Neither ε1 nor ε2 determinants in the Fc portion of ε-chains were degraded by this treatment.
Studies on Leukocyte Migration Inhibition: The Role of E Rosette-Forming Cells in Specific Antigen-Induced Inhibition of Migration
Authors: Fimmel Peter J.
Published: December 2022

The role of E rosette-forming cells in leukocyte migration inhibition was investigated with human buffy coat cells fractionated by combined E rosette formation and Ficoll-Hypaque (FH) density gradient centrifugation. The migration of unfractionated dextran-sedimented leukocytes and FH-separated mononuclear cells (MN) were equally inhibited in the presence of Escherichia coli somatic antigen (E. coli), purified protein derivative of tuberculin (PPD) and influenza virus antigen (Flu). Polymorphonuclear leukocytes (PMN) separated from whole leukocytes by one FH density gradient fractionation contained > 1.0% E rosette-forming cells and were inhibited by E. coli and PPD. PMN submitted to three transits of FH (DF III) contained ≤ 0.02% E rosette forming cells and were unaffected by each of the three antigens. The migration of mixtures of 10% MN and 90% DF III cells were inhibited in the presence of specific antigen, while MN depleted of E rosette forming cells (≤ 0.5%) pooled with equal numbers of DF III cells did not respond to any antigen by migration inhibition. These results show that PMN purified by three transits of FH are unable to respond to specific antigen by inhibition of migration from microcapillary tubes, and specific antigen-induced inhibition of migration of peripheral blood leukocytes does not occur in the absence of E rosette-forming cells in normal subjects.
Studies on the Mechanism of Lymphocyte-Mediated Cytolysis
Authors: Bubbers J. Eric , Henney Christopher S.
Published: December 2022

The lysis of DBA/2 mastocytoma cells (P815) by alloimmune C57BL/6 thymus-derived lymphocytes was inhibited by cytochalasin A (CA) and cytochalasin B (CB). Although CB prevented incorporation of glucose into lymphocytes, the effector cell susceptibility to the drug was independent of the hexose content of the culture medium. Indeed, cytolysis occurred in the absence of exogenous hexose. Furthermore CA which, like CB, inhibited the adsorption of effector spleen cell populations onto homologous allogeneic monolayers and prevented the migratory movement of lymphoid cells did so without affecting incorporation of glucose into lymphocytes.

These findings suggest that the inhibition of cytolysis by CA and CB is mediated by common inhibitory effects on membrane mobility rather than by affecting glucose uptake.
Demonstration of a Tumor-Associated Surface Antigen in Marek's Disease
Authors: Witter R. L. , Stephens E. Ann , Sharma J. M. , Nazerian K.
Published: December 2022

Surface antigenic markers were detected on three classes of Marek's disease (MD) tumor cells, i.e., MD lymphoma cells, cultured cells of the MSB-1 lymphoblastoid cell line, and JMV lymphoblastic leukemia cells, by indirect membrane immunofluorescent staining with serum from chickens immunized with JMV cells or from rabbits immunized with MSB-1 cells. This surface antigen was not detected on normal chicken lymphocytes, RPL-16 tumor cells (tranedormed by an avian RNA virus, or MD virus-infected fibroblasts that were positive for viral membrane antigen (MA). Furthermore, the surface antigen appeared unrelated to embryonic or histocompatibility antigens. This antigen is provisionally designated as a Marek's disease tumor-associated surface antigen (MATSA). The MATSA's on JMV, MSB-1 and MD lymphoma cells were related but not identical as demonstrated by antiserum titration, absorption and blocking tests with homologous and heterologous systems.
Blocking of Spleen Cell Activity against Target Mammary Tumor Cells by Viral Antigens
Authors: Blair Phyllis B. , Lane Mary-Ann , Yagi Mary Jane
Published: December 2022

Spleen cells from BALB/c females exposed to or neonatally infected with mammary tumor virus (MTV) are cytotoxic to MTV-induced mammary tumor cells in microcytotoxocity assay. This activity can be partially or completely blocked by pretreatment of spleen cells with MTV purified from milk. Murine leukemia virus (MuLV) has no effect. T cell responses of virgin and multiparous BALB/cfC3H females are effectively blocked. Non-T cell responses of multiparous BALB/cfC3H females or of virgin BALB/c females are blocked by some but not all of the MTV antigen preparations. MuLV, but not MTV, can block activity of spleen cells from MuLV-sensitized donors against target MuLV-producing tumor cells.
Antigenicity of Carcinogen and Viral Induced Sarcomas in Inbred and Random Bred Guinea Pigs
Authors: Forni Guido , Rhim Johng S. , Pickeral Susan , Shevach Ethan M. , Green Ira
Published: December 2022

A tumor specific transplantation antigen (TSTA) has been detected in a methylcholanthrene (MCA) induced guinea pig tumor. It was possible to induce resistance to rechallenge with the tumor by immunization with irradiated cells in CFA. In contrast, the same technique failed to detect TSTA in two viral (Kirsten strain mouse sarcoma virus, Ki-MSV) induced guinea pig tumors; these results are similar to observations made with mouse Ki-MSV-induced tumors. Transplantation studies with these tumors in both inbred and random-bred guinea pigs showed a complexity of growth and rejection patterns. The B alloantigen, a major serologically defined antigen of the guinea pig histocompatibility complex, seemed to play a central role in acting as a guinea pig transplantation antigen. In all cases studied, the absence of B antigens in the recipient led to tumor rejection and anti-B antibody protection.
Isolation and Characterization of the Sheep Erythrocyte Receptor in Man
Authors: Pyke Kenneth W. , Rawlings Gayle A. , Gelfand Erwin W.
Published: December 2022

Properties of the sheep erythrocyte receptor on human lymphocytes were examined from two different aspects: inhibition of E-rosette formation by antisera, and extraction of T and non-T cells with 3 M KCl. Antisera to several T cell populations produced significant inhibition of E-rosette formation by human peripheral blood lymphocytes, whereas anti-B cell, anti-β2-microglobulin, anti-I, and anti-i were ineffective. Sera with activity to HL-A subgroups on the HPBL tested produced complete rosette inhibition, although nonspecific anti-HL-A produced no inhibition. T cell preparations extracted by a 3M KCl technique contained activity that could be bound to SRBC and produce inhibition of rosette formation by HPBL. This inhibitory component could be quickly eluted from the SRBC and retain its inhibitory activity. No inhibitory activity could be demonstrated in extracts of B cells and non-lymphoid cells. These results suggest that 1) T cell-specific and HL-A specific antisera inhibit E-rosette formation; 2) β2-M is not involved in the SRBC receptor as an active component; and 3) the receptor may be isolated by 3 M KCl extraction, and partially purified by the use of SRBC as a specific immunoadsorbent, without loss of biologic activity.
Lymphocyte Abnormalities in Aleutian Disease Virus Infection of Mink: Decreased T Lymphocyte Responses and Increased B Lymphocyte Levels in Persistent Viral Infection
Authors: Perryman Lance E. , Banks Keith L. , McGuire Travis C.
Published: December 2022

Persistence of virus and a marked hypergammaglobulinemia are consistent features observed in mink with Aleutian disease virus infection. These events may result from altered lymphocyte function in infected mink. To test this possibility we determined the ability of lymphocytes from infected mink to respond to phytolectins (PHA and Con A), and a specific antigen (KLH) in a lymphocyte transformation test and quantitated the number of lymphocytes bearing surface immunoglobulin. Infected mink had a significantly lower response than normal mink to PHA and Con A (p < 0.025), and the difference in the responses increased progressively with time post-infection. Infected mink immunized with KLH also had a decreased ability to respond to this antigen when compared to normal immunized mink (p < 0.001). The number of lymphocytes bearing surface immunoglobulin was significantly elevated in three of 10 mink infected for 10 weeks, and in all of seven mink infected 32 weeks (p < 0.001).
Naturally-Ocurring Low Molecular Weight IgM in Patients with Rheumatoid Arthritis, Systemic Lupus Erythematosus, and Macroglobulinemia
Authors: McDougal J. S. , Harisdangkul Valee , Christian C. L.
Published: December 2022

Some physicochemical properties of naturally occurring low molecular weight IgM (LMW IgM), IgM and their subunits obtained by reduction-alkylation were compared. On the basis of analytical ultracentrifugation, SDS polyacrylamide electrophoresis, analytical Sephadex chromatography, and heavy (H) and light (L) chain composition purified LMW IgM had an S20, w constant of approximately 8S, a molecular weight of 195,000 to 205,000 and a structural formula H2L2. Reduction and alkylation of IgM (IgM RA) by two techniques produces two components which could be distinguished by analytical ultracentrifugation, sucrose density ultracentrifugation, and gel filtration studies. IgM RA-cys had an estimated molecular weight similar to LMW IgM and IgM-2ME had physical properties similar to IgG. The molecular weights of H and L chains of IgM and LMW IgM were similar: 69,000 to 75,500 and 26,000 to 27,000, respectively. Unlike LMW IgM and IgM RA-cys, the less dense IgM RA reduced with 2ME dissociated into half subunits, and free H and L chains in SDS gel electrophoresis, indicating that it lacked intra-subunit disulfide bonds. J chain was not detected in LMW IgM or in the chromatographically purified reduced-alkylated subunits from a patient whose IgM contained J chain.

Carbohydrate analyses of IgM, LMW IgM, and reduced-alkylated IgM products did not demonstrate differences in individual sugars or in the percentage of total carbohydrate.

The different sedimentation and gel filtration properties of the two components obtained by reduction-alkylation of IgM could not be attributed to variations in peptide chain or carbohydrate content, rather they are probably dependent on conformational changes resulting from variable reduction of intra-subunit disulfide bonds.
Early Steps in Specific Tumor Cell Lysis by Sensitized Mouse T Lymphocytes
Authors: Martz Eric
Published: December 2022

Addition of high molecular weight dextran to culture medium prevents the initiation of T lymphocyte-mediated killing by holding the cytolytic T lymphocytes (CTL) and target cells in suspension and preventing intercellular contact. Suspension in 10% dextran was used to interrupt the ongoing formation of adhesions between CTL and target cells already in contact in a centrifuged pellet. The results demonstrate that 1) firm adhesions form between CTL and target cells within 1 min at 37°C; 2) once formed, these adhesions are stable at low temperature and are resistant to mechanical shearing forces; 3) these adhesions can be disrupted by EDTA; 4) immediately after the adhesions form, separation of the CTL from the target cells prevents lysis of the latter; 5) after incubation of targets adhering to CTL for an additional 6 min at 37°C, removal of the CTL no longer prevents target cell lysis. Thus, target cells become “programmed” for subsequent lysis within a few minutes after contact with CTL, after which lysis occurs during the next several hours without further participation of the effector cell.

At 15°C, adhesions form 1/17 as fast as at 37°C. Programming of target cells for lysis occurs 1/76 as fast at 15°C as at 37°C. Thus, the programming for lysis step is about 4-fold more temperature dependent than the adhesion step.

In addition to being detected by subsequent target cell lysis in 10% dextran, the adhering cell clusters can be counted with low power microscopy. This permitted verification that EDTA separates the clusters after programming for lysis is complete. Moreover, the great majority of the clusters seen at 37°C are antigen-specific. Knowledge of the cluster size distribution and the subsequent level of lysis permits the deduction that not less than 6% of the sensitized peritoneal cell populations used were CTL.
Immunochemical Characterization of the Anti-RNA Antibodies Found in Scleroderma and Systemic Lupus Erythematosus
Authors: Alarcón-Segovia Donato , Fishbein Eugenia
Published: December 2022

In a previous study, all 40 sera from patients with scleroderma, 20 of 40 sera from SLE patients, but none of 40 sera from normal controls, were found to have antibodies to ssRNA. All scleroderma sera were also found to react with HSA-coupled uridine and UMP and their reaction with ssRNA could be inhibited by uracil, uridine, and UMP.

To characterize further these uracil-specific anti-RNA antibodies found in scleroderma and compare them with the anti-RNA antibodies found in SLE, we tested their reactivity with Poly (U) and with Poly (A)·Poly(U) in CIE. All 40 scleroderma sera reacted with Poly (U) and all but one failed to react with Poly (A)·Poly(U). This same serum was the only one in which the reaction with Poly (U) could not be inhibited with uracil.

Reactivity of SLE sera was strikingly different from that found in scleroderma sera. Seventeen of 34 SLE sera studied reacted with ssRNA but only four of these reacted with Poly (U). Conversely, two SLE sera that reacted with Poly (U) did not react with ssRNA. Fifteen reacted with Poly (A)·Poly (U) and only two of these failed to react with ssRNA. Five SLE sera which were reactive with ssRNA did not precipitate with Poly (A)·Poly (U).

All SLE sera which reacted with Poly (U) could be inhibited with uracil, although less effectively than in scleroderma. Reactivity with Poly (A)·Poly (U) was not inhibited with uracil nor with adenosine.

These findings confirm that antibodies to RNA that are found in scleroderma are directed to uracil and thus specific to ssRNA, whereas RNA antibodies found in SLE sera are heterogeneous and directed to either the base, to the site of union of the base and sugar moiety to the ribose backbone, or to the helical structure of double stranded RNA. These differences and the respective antigenic specificities of these anti-RNA antibodies found in scleroderma and SLE may be theoretically important.
Relationship between Immunoglobulin-Bearing Lymphocytes and Cells Reactive with Sensitized Human Erythrocytes
Authors: Kurnick James T. , Grey Howard M.
Published: December 2022

Peripheral blood lymphocytes were evaluated for surface immunologlobulin and the ability to form rosettes with anti-Rh coated erythrocytes (EA). Ninety percent of the EA-rosetting cells also possessed surface IgG. This surface IgG was mostly lost after a 1-hr incubation at 37°C. These data are discussed in relation to the work of Fröland and Natvig and Lobo, et al., and the suggestion is made that the EA-rosette assay detects a population of cells with a very avid Fc receptor that bears cytophilic IgG on its surface.
Evidence for Cell-Receptor Activity in Lymphocyte Stimulation by Staphylococcal Enterotoxin
Authors: Warren John R. , Leatherman Dennis L. , Metzger Joseph F.
Published: December 2022

A wide dose-response curve and the inhibitory effect on mitogenicity of specific antitoxin suggest that polyclonal lymphocyte activation by staphylococcal enterotoxin requires direct interaction of toxin with lymphocyte receptors of low avidity for the protein. Staphylococcal enterotoxins A, B, and C1 demonstrated equivalent mitogenic activity. Lymphocyte receptors involved in enterotoxin activation thus appear to be specific for nonantigenic regions of the toxin molecule. Monosaccharide (hapten) inhibition data indicate that lymphocyte receptors for staphylococcal enterotoxin lack α-mannoside, galactose, acetylgalactosamine, acetylglucosamine, and fucose (or closely related saccharides) as determinant sugars and thus differ significantly in structure from lectin cell receptors.
Presidential Address to the American Association of Immunologists, Delivered in Atlantic City, New Jersey, April 17, 1975
Authors: Kunkel Henry G.
Published: December 2022

Each year it has been the privilege of the President of the Association to make a few remarks to the membership. Frequently this has been a scientific presentation, but it also has taken many other forms. Last year, when I found Baruj Benacerraf vascillating on the question as to which path he should take, I urged him to seize the opportunity and give us some of his more philosophical thoughts. We had heard enough about Ir genes. I stated that I would certainly follow his example next year. He, of course, then did this very successfully. In my own case, however, as next year drew ever closer, I had some second thoughts. It suddenly occurred to me that a scientific talk published in the Journal of Immunology would be a valuable asset. I would have to be sole author on the paper, something that hasn't occurred in many years and this would be quite impressive with my grants soon coming up for renewal.
IgG Antibody Suppression of the IgM Antibody Response to Toxoplasma Gondii in Newborn Rabbits
Authors: Araujo Fausto G. , Remington Jack S.
Published: December 2022

Studies were performed in an experimental rabbit model in an attempt to explore the possibility that the absence of an IgM antibody response in human infants congenitally infected with Toxoplasma may be caused by a suppressive effect of IgG antibody passively transferred from the mother. The IgM antibody response to Toxoplasma gondii infection in newborn rabbits previously injected with IgG Toxoplasma antibody was significantly suppressed as determined by an immunofluorescent antibody assay (IgM-IFA). In newborn rabbits infected with Toxoplasma, IgM-IFA antibody was first demonstrated 15 days after infection and reached a peak titer of 1:128 in two animals on days 20 and 25. In newborn rabbits injected with IgG Toxoplasma antibody and then infected with Toxoplasma, the IgM-IFA antibody response was considerably lower; the highest titer was 1:16 and occurred 25 and 30 days after infection. The results of these studies lend support to the concept that maternally transmitted IgG may suppress the IgM antibody response in the fetus and newborn infant.
Cytotoxic Activity of Lymphocytes
Authors: Reed William P. , Lucas Zoltan J.
Published: December 2022

Macrophage cell-mediated cytolysis (M-CMC) of tumor cells may be measured by culturing target cells on macrophage monolayers and determining target cells on macrophage monolayers and determining target cell survival by either 86Rb or 3H-thymidine incorporation. Using these techniques, one can demonstrate cytotoxicity of non-immune human and rat macrophages to a variety of tumor cell lines. Toxicity becomes maximal 8 to 16 hr after attaching macrophages to plastic surfaces and disappears after 36 to 48 hr. Addition of Escherichia coli endotoxin at 48 hr restores macrophage toxicity. Culture of macrophages on plastic surfaces also results in the release, non-specifically, of a soluble cytotoxin whose kinetics of release, disappearance, and reappearance after endotoxin all parallel the M-CMC. The amount of toxin released into cultures containing target cells is markedly reduced, suggesting that it is the mediator of the M-CMC and that it is utilized or adsorbed during target destruction. Sephadex G-100 chromatography and treatment with heterologous anti-lymphotoxin can distinguish this toxin from lymphotoxin.
Receptors for Antigen on Lymphoid Cells
Authors: Greeley Elizabeth A. , Scott David W.
Published: December 2022

The nature of specific adherence of rat anti-TNP PFC to TNP-GRBC has been investigated with PLL-fixed antigen monolayers as cellular immunoadsorbents and as plaque indicators. The immunoglobulin nature of the molecule responsible for PFC adherence is suggested by the fact that pretreatment of the PFC with rabbit anti-rat Ig antisera, but not anti-histocompatibility antisera, inhibits adherence. Removal of the adherence capacity of early PFC with the proteolytic enzymes papain and pronase, or by “capping” with anti-Ig is followed by slow regeneration of the ability to adhere, suggesting that adherence is due to membrane rather than secreted immunoglobulin, the latter being detectable within minutes after enzyme treatment. Several time-related events relating to PFC adherence were observed. 1) Both direct and indirect PFC are capable of specific adherence; the ability to adhere, however, tends to decline with time, especially after secondary immunization. 2) Although early PFC adherence is unaffected by trypsin treatment, later populations become increasingly sensitive. 3) Pretreatment of PFC at various times after primary immunization with antisera specific for rat µ-chain indicates that IgM and possibly early IgG-secreting PFC have µ heavy chains on their surface. These data suggest that the PFC membrane is progressively changing during the maturation of the antibody response.
Differential Susceptibility of Rabbit sIgA Subclasses to Trypsin Cleavage: Characterization of the Fragments Obtained from the g Subclass
Authors: Tseng Jeenan , Knight Katherine L.
Published: December 2022

Rabbit secretory IgA (sIgA) was digested with trypsin at 37°C for 30 min and four fractions were isolated by gel filtration. These fragments were characterized by ultracentrifugation, and by their antigenic properties as undigested sIgA, Fab2α and Fc2α. (Two Fc2α fragments differing in their content of secretory component were obtained.) The IgA-f subclass molecules were resistant to cleavage and were found in the undigested sIgA fraction; the IgA-g subclass molecules were cleaved into Fc2α and Fab2α fragments. The g allotypic determinants of IgA-g molecules were found on both the Fc2α fragments and the Fab2α fragments. The Fc2α and Fab2α fragments obtained from trypsin-digested sIgA were compared by means of antigenic properties and peptide maps with the Fc2α and Fab2α fragments obtained from papain-digested sIgA; no differences attributable to the α-chain were found. Thus, papain and trypsin cleave the α-chain of IgA-g molecules at similar but not necessarily identical postions.
Lysis of Tumor Cells by Antibody and Complement
Authors: Boyle Michael D. P. , Ohanian Sarkis H. , Borsos Tibor
Published: December 2022

The increase in susceptibility to killing by rabbit antibody and guinea pig complement of guinea pig hepatoma cells (line-10), after treatment with certain metabolic inhibitors, did not correlate with the mobility of antigen on the cell surface as measured by indirect immunofluorescence.
The Identification of a Previously Unrecognized Subcomponent of the First Component of Complement
Authors: Assimeh Seth N. , Painter Robert H.
Published: December 2022

The use of an affinity chromatography method designed to isolate C1 from serum has led to the discovery of a novel plasma protein, II-P2, associated with C1. The persistent Ca++-dependent association of II-P2 with C1 subcomponents following euglobulin precipitation, affinity chromatography on Sepharose-IgG, and density gradient ultracentrifugation indicates that II-P2 might be a C1 subcomponent. Using purified preparations of II-P2 it was found that a) II-P2 was retained on Sepharose-IgG through a Ca++-dependent link with C1q, b) II-P2 enhanced the C1 activity of mixtures of C1s and C1q in a dose-dependent fashion, c) II-P2 bound firmly to EAC1q4 cells and enhanced their C1s-binding ability. Fractionation of C1 by DEAE-Cellulose chromatography under the conditions that led to the original identification of C1q, C1r, and C1s resulted in recovery of II-P2 in the fractions containing C1r. The evidence presented confirms that II-P2 is a C1 subcomponent (C1t).
The Macromolecular Structure of the First Component of Complement
Authors: Painter Robert H. , Assimeh Seth N. , Painter Robert H.
Published: December 2022

The binding of C1 to IgG and the interactions between C1 subcomponents have been studied by affinity chromatography of serum C1 and purified C1 proteins on Sepharose-IgG. Affinity chromatography of serum on Sepharose-IgG resulted in the binding of C1; subsequent washing with EDTA removed only C1s and C1t. Affinity chromatography of serum-EDTA on Sepharose-IgG resulted in binding of only C1q and C1r. Affinity chromatography of serum on Sepharose-tryptophan-modified-IgG resulted in the binding of only C1r and C1s. By the use of purified C1 proteins and Sepharose-IgG in binding studies it was confirmed that both C1q and C1r bind independently to sites on IgG and hold C1s and C1t by Ca++-dependent bonds. Measurements of the hemolytic activity of various combinations of C1 subcomponents confirmed the data obtained by the affinity binding studies. Both C1t and C1r independently enhanced the C1 activity of C1q-C1s mixtures; maximal activity required all four subcomponents. Sucrose gradient ultracentrifugation of mixtures of C1 proteins showed formation of the following complexes: C1qs (12S), C1qrs (15S), C1qst (18–23S), and C1qrst (19S).

The evidence suggests that the spatial sequence of the components of the Sepharose-IgG-Serum C1 complex is: Sepharose-IgG: C1q: C1t: C1s: C1r: IgG-Sepharose. The probable physiologic significance of this model is discussed.
Interaction Between Lymphocytes and Inflammatory Exudate Cells
Authors: Yoshinaga Masaru , Nakamura Shuji , Hayashi Hideo
Published: December 2022

Thymocytes of C3H/He mice were poorly triggered to induce DNA synthesis by phytohemagglutinin but fairly well triggered when they were mixed with small number of inflammatory peritoneal exudate polymorphonuclear leukocytes or macrophages, or with the soluble product(s) of these cells.

Peritoneal exudate cells were mainly harvested at 3 hr, largely consisting of polymorphonuclear leukocytes, and at 96 hr, largely consisting of macrophages, after intraperitoneal injection of 0.2% sodium caseinate. Addition of 22 to 26 × 103 of 3-hr exudate cells, 22 to 24 × 103 of 48-hr exudate cells, or 22 to 24 × 103 of 96-hr exudate cells to syngeneic thymocyte culture (3 × 106) resulted in marked enhancement of DNA synthesis by thymocytes stimulated with 1 µ1 PHA. Similar enhancement of DNA synthesis was also observed in the presence of 200 µ1 of cellfree supernatant fluid of peritoneal exudates of 3 hr or 9 hr after casein stimulation. Conversely, two hundred microliters of 0-hr, 48-hr, or 96-hr peritoneal fluid showed a slight suppressing effect on thymocyte response and 50 to 100 µl of these exudates showed no enhancing effect. Furthermore, 50 to 800 µ1 of culture supernatants from 1.6 × 106 cells/4 ml of 3-hr exudate cells, 96-hr exudate cells, or purified macrophages also caused a similar enhancement of DNA synthesis by thymocytes. On the other hand, excess numbers of exudate cells (27 to 28 × 103 of 3-hr exudate cells, or 25 to 29 of 96-hr exudate cells) or excess amounts of culture supernatants (800 µl from 6.4 to 12.8 × 106/4 ml of 3-hr cells, or 200 µl from 12.8 × 106/4 ml, 400 µl from 6.4 × 106/4 ml, or 800 µl from 3.2 × 106/4 ml of 96-hr cells) diminished the enhancing potency on thymocytes. The cooperative function by polymorphonuclear leukocytes or by macrophages on the lymphocyte response was thus postulated.
Cytoplasmic Immunofluorescence of Bone Marrow Plasma Cells Producing Immunoglobulins of the Four IgG Subclasses
Authors: Morell Andreas , Skvaril Frantisek , Hijmans Willy , Scherz Rudolf
Published: December 2022

Immunofluorescence studies were performed on bone marrow samples from 21 individuals with normal or slightly elevated serum IgG levels. On the average, 40.7% of the IgG-positive plasma cells contained molecules of the subclass IgG1; 30.6% had intracellular IgG2; 21.7%, IgG3, and 7.0%, IgG4. These figures were compared to the relative serum IgG subclass concentrations. The principal finding was the discrepancy between the high relative number of IgG3-producing bone marrow plasma cells and the low relative IgG3 serum concentration (5.5% of the total IgG). It was explained by the short biologic half-life of molecules of this subclass.
Concanavalin A Preparations with Activities Related to Bacterial Lipopolysaccharide
Authors: Brunson Kenneth W. , Watson Dennis W.
Published: December 2022

Concanavalin A. (Con A)2 and other plant lectins have recently been extensively employed as biochemical probes in order to investigate cell surfaces and specific binding sites on these surfaces (1–3). Con A is also a potent lymphocyte mitogen (4, 5). In the course of comparative investigations of the mitogenic property of Con A and bacterial mitogens, we found Con A preparations to be pyrogenic. Pyrogenicity of Con A had previously been reported (6); however, the febrile response in American Dutch rabbits we observed was strikingly similar to the biphasic curves obtained upon i.v. injection of Gram-negative bacterial endotoxin (lipopolysaccharide). Moreover, by using dose response relationships of several Con A preparations observed in rabbit pyrogen tests, the pyrogenic activity could be biologically quantitated, as reported for endotoxin (7). An amount necessary for a minimal pyrogenic dose at 3 hr (MPD-3) could thus be determined (7). We therefore felt compelled to investigate further any possible cross-reactive relationships between the Con A preparations and endotoxin.
Lymphocyte-Mediated Cytotoxicity against Influenza Virus-Infected Cells: An in Vitro Method
Authors: Greenberg S. B. , Criswell B. S. , Couch R. B.
Published: December 2022

A direct test of cell-mediated function is described using a 51Cr release assay for lymphocyte-mediated cytotoxicity (LMC) of influenza virus- (H3hkNeq1) infected tissue culture cells. LMC was detected only in volunteers with serum-neutralizing antibody to H3hkNeq1 influenza virus. A positive correlation between degree of LMC and serum neutralizing antibody titers was found in the tested volunteers.
Immunopathology of Mouse Hepatitis Virus Type 3 Infection
Authors: Le Prévost C. , Virelizier J. L. , Dupuy J. M.
Published: December 2022

Three types of viral sensitivity were observed in various mouse strains upon MHV3 infection: resistance, full susceptibility, and semisusceptibility. In the latter type, seen in several inbred strains including C3H, approximately 50% of the adult injected animals resisted to the acute disease. Most of the surviving mice, however, developed a chronic disease with a wasting syndrome and occurrence of paralysis. The chronic period of the disease was characterized by a persistent viral infection, since MHV3 virus was recovered from brain, liver, spleen, and lymph nodes throughout the evolution in most of the animals. In addition, a correlation was observed between the clinical evolution and the titer of virus tested 4 days after infection.
B Lymphocyte Differentiation Induced by Lipopolysaccharide
Authors: Kearney John F. , Lawton Alexander R.
Published: December 2022

Cultures of mouse fetal liver and spleen, stimulated by bacterial lipopolysaccharide, gave rise to plasma cells staining for IgM, IgG2, IgG1, and IgA. Cells containing IgM and IgG2 were found in cultures from 17-day fetuses, coincident with the appearance of B lymphocytes bearing cell-surface IgM. IgG1- and IgA-containing cells were induced in cultures from 19-day fetuses and 1-day-old mice. The capacity to give rise to immunoglobulin-secreting cells of all classes preceded the development of a significant proliferative response to LPS; the proportions of cells staining for each class reached adult values by 1 day of age whereas the proliferative response did not mature until 3 weeks.
The Immune Response in the Hamster
Authors: Portis J. L. , Coe J. E.
Published: December 2022

Hamster 7S IgG1 and IgG2 antibodies to hen-egg albumin (HEA) were tested for their capacity to bind to macrophage cytophilic Ig receptors. Both IgG1 and IgG2 were cytophilic for hamster macrophages though the membrane receptor had a predominant specificity for IgG1. Hamster IgG1 bound primarily to homologous macrophages whereas IgG2 bound to macrophages from other rodent species as well. The binding of hamster Ig to hamster macrophages was inhibited by a wide range of heterologous rodent sera. The only exception was guinea pig serum since guinea pig IgG2 was found to bind only to homologous macrophages. Sheep red blood cells (SRBC) coated with hamster IgG2 were ingested by macrophages more readily than those coated with hamster IgG1. Thus, there appeared to be a paradoxical relationship between the apparently strong affinity of IgG1 for the hamster macrophage Ig receptor and its relatively weak ingestion promoting activity. Implications of this phenomenon are discussed.
Immunogenetic Analysis of H-2 Mutations
Authors: Klein Jan , Forman James , Hauptfeld Věra , Egorov Igor K.
Published: December 2022

Mutation M523 (H-2ka) occurred spontaneously in strain CBA/CaLacSto and was discovered during routine skin graft testing for genetic homogeneity. By linkage and complementation tests, the mutation was previously mapped in the K end of the H-2 complex. We demonstrate that the mutation occurred in the K region, without affecting the I region in the K end of the complex. The mutant antigens cause rejection of skin grafts, stimulate cells in mixed lymphocyte culture, and function as stimulators as well as targets in cell-mediated lymphocytotoxicity. Yet, they are serologically indistinguishable from the antigens of the original strain and do not induce formation of humoral antibodies upon immunization of the CBA strain. Together with the results obtained on testing of other H-2 mutants, the data strongly support the notion that classical H-2 antigens (i.e., products of the H-2K and H-2D loci) can function as lymphocyte-stimulating determinants, and that I-region differences are not required for the induction of strong cell-mediated lymphocytotoxicity.
Immune Mechanisms in Leukemia: Suppression of Cellular Immunity by Drugs and X-Irradiation
Authors: Martinez Douglas , Lukasewycz Omelan A. , Murphy William H.
Published: December 2022

The relative suppressive effects of x-irradiation (XR), cyclophosphamide (CY), prednisolone (PRD), and methotrexate (MTX) on the primary and secondary cellular immune response of C58/wm mice to syngeneic line Ib transplantable leukemia (Ib cells) were quantified. An LD10 dose of each agent was used for immunosuppression. XR, CY, and PRD were markedly suppressive for the primary immune response if given 24 hr before mice were immunized to Ib cells but less immunosuppressive if given 24 hr later. MTX was only slightly immunosuppressive. XR, CY, and PRD also suppressed the secondary immune response if given before but not after antigen. The immunosuppressive effect of these agents was evaluated by defining their median immunosuppressive dose or the median time in days required for mice to recover from graded doses of each immunosuppressive agent. For example, the median recovery time from an LD10 of XR, CY, and PRD was 29.3, 19.7, and 3.7 days, respectively. Immunologic competence remaining after XR or drug treatment was quantified in terms of the LD50 dose of Ib cells required to kill recipient mice. For XR, CY, PRD, and MTX it was 106.16, 102.15, 106.90 and τ; 107.0 viable Ib cells, respectively. The overall results provided evidence that the primary and secondary cellular immune responses to a weak syngeneic tumor antigen were resistant to immunosuppression once they were initiated. There was a good correlation between the relative immunosuppressive effect of the test agents and the amount that they reduced the number of immune spleen cells. The agents also impaired the immunocompetence of individual spleen cells. Mechanisms by which XR or drugs might exert their immunosuppressive effects were discussed.
Evidence that Uridine Incorporation Is not a Selective Marker for Mouse Lymphocyte Subclasses
Authors: Gutman George A. , Weissman Irving L.
Published: December 2022

We demonstrate here that mouse T+ and Ig+ lymphocytes do not show the differential incorporation of 3H-uridine into acid precipitable cellular components described for these lymphocyte subpopulations in the rat.
Macrophages Activated as Suspension Cultures with Lymphocyte Mediators Devoid of Antigen Become Cytotoxic for Tumor Cells
Authors: Churchill W. Hallowell , Piessens Willy F. , Sulis Carol A. , David John R.
Published: December 2022

Normal peritoneal exudate cells (PEC) were activated as suspension cultures either in mediator-rich supernatants from o-chlorobenzoyl-bovine γ-globulin (OCB-BGG) stimulated lymphocytes or in antigen-free Sephadex fractions from these supernatants. After 24 hr incubation the activated macrophages showed enhanced cytotoxicity for line 1 hepatoma and were inhibited in migration. The adherent cell fractions of PEC, recovered by trypsinization from monolayers and activated by this technique, were as cytotoxic as unfractionated PEC. Lymphocyte supernatants and antigen-free fractions of the supernatants induced comparable macrophage-mediated tumor cytotoxicity. Treatment of activated macrophages with trypsin did not alter their cytotoxic capacity.
Further Studies on the Biologic Properties of Guinea Pig IgG1 Antibodies
Authors: Vuagnat Philippe
Published: December 2022

Normal rats were injected with guinea pig anti-rat glomerular basement membrane antibodies of the IgG1 or IgG2 class or with their F(ab′)2 fragment, in order to study which antibody site triggers the alternate complement pathway in vivo. Both IgG classes were able to induce a heavy proteinuria and led to C3 deposition in the glomeruli in a pattern similar to their own distribution along the glomerular basement membrane, as shown by the immunofluorescence technique. The Fab(ab′)2 fragment of IgG2 did not produce C3 binding or proteinuria. The F(ab′)2 fragment of IgG1 was difficult to obtain devoid of Fc determinants. A F(ab′)2 fragment of IgG1 still bearing Fc determinants led to C3 binding and proteinuria, whereas the true F(ab′)2 fragment of IgG1 had none of these effects in two out of three animals.
Determination of Antibody to Hepatitis B Core Antigen by Means of Immune Adherence Hemagglutination
Authors: Tsuda Fumio , Takahashi Takashi , Takahashi Kazuaki , Miyakawa Yuzo , Mayumi Makoto
Published: December 2022

A simple and rapid method utilizing immune adherence hemagglutination has been developed for the detection of antibodies to hepatitis B core antigen (anti-HBc). Hepatitis B core antigen (HBcAg) was prepared from Dane particles that had been isolated from plasma of asymptomatic antigen carriers. The method was specific and about 10 times more sensitive than the conventional complement-fixation method.

A total of 215 serum samples obtained from healthy blood donors were surveyed for HBcAg and anti-HBc, as well as for hepatitis B surface antigen (HBsAg) and antibody to HBsAg (anti-HBs). Anti-HBc was found in 36 serum samples, at a prevalence rate higher than that of anti-HBs (31/215). Anti-HBc in 28 serum samples was associated with anti-HBs, and in two it was found with HBsAg. There were six serum samples in which anti-HBc was detected without any of anti-HBs, HBcAg, and HBsAg.
Activated Macrophages in Congenitally Athymic “Nude” Mice and in Lethally Irradiated Mice
Authors: Cheers Christina , Waller Robyn
Published: December 2022

Resistance to the facultative intracellular bacteria, Brucella abortus and Listeria monocytogenes, is principally the result of acquisition of enhanced antibacterial activity by host macrophages, probably in response to lymphokines released by T lymphocytes. However, the present paper describes a surprisingly high resistance on the part of both congenitally athymic “nude” mice and of lethally irradiated mice compared with normal controls. This enhanced bactericidal activity was evident 24 hr after infection, and could also be demonstrated in macrophages from nude mice cultured in vitro. It was concluded that the macrophages of these animals had been activated before infection.
RNA and Protein Synthesis in Spontaneous Rosette Formation by T Lymphocytes
Authors: Bushkin Sheila C. , Pantic Vojislav S. , Good Robert A.
Published: December 2022

Formation of rosettes with sheep red blood cells (SRBC) by peripheral blood lymphocytes and thymocytes has been studied. Trypsinization of these cells abolishes the capacity to form SRBC-rosettes. The process of re-establishment of this capacity to form SRBC-rosettes, in vitro, has been shown by use of metabolic inhibitors to require transcription, translation, and protein synthesis. These observations are interpreted in terms of the T cell markers as the expression of a definitive process or step in lymphoid cell differentiation.
Studies on Nonidet P40 Lysis of Murine Lymphoid Cells
Authors: Hart David A.
Published: December 2022

Lymphoid cells from A/J mice were iodinated (125I) by the lactoperoxidase method and lysed with the non-ionic detergent NP-40. The plasma membrane glycolipid receptor for cholera toxin and cell surface immunoglobulin were utilized in immune precipitation systems to characterize the degree of dissociation of the plasma membrane under various conditions. It was found that at 0.1% NP-40 and at cell concentration from 5 to 10 × 107 cells/ml, lipid-protein and protein-lipid-protein complexes formed in NP-40 which were soluble after centrifugation at 105 × G. Column chromatography of 125I-cell lysates on agarose A-0.5 M in 0.1% or 0.5% NP-40/PBS indicated that the majority of iodinated cell surface material existed as aggregates in detergent micelles. The availability of the oligosaccaride moiety of the glycolipid to interact with the cholera toxin was dependent on both the detergent concentration and the cell concentration used for cell lysis. However, the cell surface immunoglobulin was immunoprecipitable under all conditions of lysis tested.
Activation of the Alternative (Properdin) Pathway by Divalent Cations
Authors: Lew Frances T. , Yukiyama Yasushi , Waks Harriet S. , Osler Abraham G.
Published: December 2022

Mg++, Mn++ and Co++ activate the C system as judged by the reduction in cobra venom factor induced hemolysis, and cleavage of properdin factor B and C3. The maximum effect requires the addition of 5 to 10 mM of these cations to dialyzed sera. Guinea pig, human and rat sera respond in similar fashion indicating that these divalent cations can trigger the alternative pathway without the addition of other activating agents.
Subfragments of Papain Solubilized TL Antigen
Authors: Stanton Thomas H. , Bennett J. Claude , Wolcott Michael
Published: December 2022

TL antigen was solubilized from the tumor ASL1 (TL. 1,2,3) by papain digestion. The subfragments of 125I-labeled TL were examined by two methods. The first involved immune precipitation followed by electrophoresis on SDS-acrylamide gels. This treatment yielded three bands of molecular weight 39,000 and 19,000, as well as material which migrated with the tracking dye. In the second procedure the papain digested material was partially purified on Sephadex G-200. The active fraction from G-200 was labeled with 125Iodine, mixed with alloantiserum and rechromatographed on G-200. The isolated immune complexes were boiled in SDS and 2-mercaptoethanol, then separated on a SDS-Sephadex G-150 column. Two radioactive peaks were eluted indicating an absence of the 19,000 m.w. component following the latter method of purification.
Regulatory Substances Produced by Lymphocytes
Authors: Namba Yuziro , Waksman Byron H.
Published: December 2022

Rat lymphotoxin (LT), a soluble mediator which kills L cells (mouse fibroblasts) within 48 hr, was partially purified by conventional protein fractionation procedures from the culture supernatants of rat lymph node cells stimulated with ovalbumin in vivo and rechallenged with the same antigen in vitro. It appeared, from the elution pattern of DEAE-cellulose chromatography, to be a neutral or slightly basic protein and was inactivated at 60°C. Its molecular weight was estimated by Sephadex gel filtration to be approximately 9 × 104 daltons. Partially purified LT, in titration studies, showed single hit kinetics. It also inhibited L cell proliferation at low concentration without significant killing effect. The heat inactivation curves of both cytotoxic and proliferation inhibitory activities were the same, suggesting that these two activities may be caused by the same molecule. However, it had almost no detectable effect on the DNA synthesis of rat lymph node cells stimulated with phytohemagglutinin (PHA), suggesting that lymphocytes, at least those which respond to PHA, are rather resistant to LT. It is suggested that activities described as “proliferation inhibitory factor” and “cloning inhibitory factor” by others may represent the activity of dilute LT. The “inhibitor of DNA synthesis” is distinct in molecular character, range of target cells, and target cell kinetics from LT.
The Effect of Thiols on Immunologic Release of Slow Reacting Substance of Anaphylaxis
Authors: Orange Robert P. , Chang Puay-Lim
Published: December 2022

Preincubation of human lung fragments with cysteine for 2.5 to 5.0 min resulted in dose-dependent, selective enhancement of the antigen-induced or anti-IgE-induced formation and release of slow reacting substance of anaphylaxis (SRS-A). Comparable effects were observed with sodium sulfide and thioglycolate but not with other more potent reducing agents or metabolites of cysteine. Sulfhydryl alkylated derivatives of cysteine were ineffective. The effects observed with the active thiols were easily reversed and could not be attributed to an action in the bioassay or on SRS-A itself. The physicochemical characteristics of the contractile activity were identical to those described for SRS-A.