bluuebunny/crossref_metadata_2025 huggingface dataset:
Macrophage Activation for Tumor Cytotoxicity: Increased Lymphokine Responsiveness of Peritoneal Macrophages During Acute Inflammation
Authors: Ruco Luigi P. , Meltzer Monte S.
Published: December 2022

Peritoneal inflammation induced by sterile irritants led to accumulation of macrophages that were more responsive to lymphokines than macrophages from resident cell populations of untreated mice. Lymphokine responsiveness was quantitated by measurement of macrophage-mediated tumor cytotoxicity induced by supernatants from antigen-stimulated immune spleen cell cultures. Tumor cytotoxicity by lymphokine-activated inflammatory macrophages was about 10-fold greater than that by equal numbers of lymphokine-treated resident cells. Analysis of the time course of activation and of lymphokine dose-response demonstrated that the increased responsiveness of inflammatory cells was a consequence of quantitative changes. Lymphokine-responsive cells in both resident and inflammatory populations were identical; the numbers of responsive cells increased with inflammation. Changes in numbers of lymphokine-responsive cells during acute inflammation were coincident with similar changes in numbers of peroxidase-positive macrophages. This finding suggested that the increased lymphokine responsiveness of inflammatory cells was dependent upon influx of young peroxidase-positive mononuclear phagocytes rather than stimulation of resident macrophages. This concept was further strengthened by the finding that whole body x-irradiation diminished the numbers of both peroxidase-positive and of lymphokine-responsive macrophages in inflammatory and resident cell populations. These data suggest that the lymphokine-responsive precursor for the activated tumoricidal macrophage during both steady-state and acute inflammatory conditions was the newly formed blood-derived mononuclear phagocyte. The 10-fold increase in numbers of lymphokine responsive macrophages in peritoneal exudate over resident cell populations reflected a similar increase in macrophage turnover induced by inflammation.
Splenic Suppressor Cells and Cell-Mediated Cytotoxicity in Murine Schistosomiasis
Authors: Coulis Paul A. , Lewert Robert M. , Fitch Frank W.
Published: December 2022

An impairment of the capacity to generate alloantigen-specific cytotoxic T lymphocytes (CTL) was observed in mixed lymphocyte cultures (MLC) established with spleen cells from mice infected with Schistosoma mansoni. This impairment, which was observed as early as the eighth week of infection, could be abrogated by the fractionation of spleen cell suspensions by the carbonyl iron/magnet method prior to the establishment of MLC. Cocultivation of normal spleen cells with increasing numbers of splenocytes from S. mansoni-infected syngeneic mice resulted in a dosage-dependent suppression of CTL generation. This “infectious suppression” was not sensitive to antiserum against mouse thymic lymphocyte antigen (MTLA).

The present studies suggest the role of a macrophage rather than a T cell as the suppressor cell in this model of cell-mediated immunity in schistosome-infected mice.
Activation of Human B Lymphocytes
Authors: Haynes Barton F. , Fauci Anthony S.
Published: December 2022

The kinetics and mechanisms of suppression of the PWM-induced PFC response of human PB lymphocytes by Con A-activated suppressor cells were investigated. It was necessary that Con A suppressor cells be present early in the preocess of activation of human B cells toward antibody synthesis, but maximal suppression of the PFC response occurred later in the culture period. In addition, Con A-activated cells, although suppressing the PFC response to PWM greater than 90% of control, did not significantly suppress the blastogenic response to PWM after 3 or 5 days in culture. On the contrary, after 3 days in culture, background tritiated thymidine incorporation as well as tritiated thymidine incorporation to PWM stimulation was increased when Con A suppressor cells are added to fresh autologous peripheral blood lymphocytes. This increased blastogenic response after three days most likely represented an autologous mixed lymphocyte reaction (MLR) of Con A suppressor cells against fresh autologous non-T cells. The induction of autoreactive cells may be one of several modes of suppression of PFC responses by Con A activated suppressor cells.
Nonimmunospecific Protein-Protein Interactions of IgG: Studies of the Binding of IgG to IgG Immunoadsorbents
Authors: Nardella Francis A. , Mannik Mart
Published: December 2022

Nonimmunospecific interactions of IgG and IgG-agarose columns were systematically studied under varying conditions. Nonimmunospecific binding to the columns was primarily due to protein-protein interactions. These nonimmunospecific protein-protein interactions of IgG were enhanced with heat-induced or chemical aggregation of IgG, low pH, low ionic strength (at pH above 4), or low temperature. Conversely, this binding was decreased with proteolytic fragmentation of IgG, high ionic strength (at pH above 4), or temperatures above 4°C. Chemical modification of IgG by acetylation, formalinization, carbamylation, or reaction with 1,2-cyclohexanedione significantly decreased these interactions. These observations suggest that above pH 4, ionic interactions caused the protein-protein binding. Below pH 4, hydrophobic interactions presumably play a major role.

These results permit the development of a rational methodology for avoiding nonimmunospecific protein-protein interactions in immunologic procedures for detection, isolation, or quantification of rheumatoid factors and other antibodies to IgG.
Plasminogen Activator of Guinea Pig Basophilic Leukocytes: Probable Localization to the Plasma Membrane
Authors: Dvorak Harold F. , Orenstein Neil S. , Rypysc Jane , Colvin Robert B. , Dvorak Ann M.
Published: December 2022

The plasminogen activator (PA) activity of guinea pig basophil-enriched leukocyte preparations was localized to basophils, and not to contaminating lymphocytes and eosinophils, by correlating PA activity with basophil frequency and, more directly, by means of an improved cytochemical method here described. PA activity was fully expressed in living cells in the absence of immunologic stimuli and was suppressed/lost to a variable extent by different techniques of cell disruption. Conversely, killed, but not living, basophils expressed significant plasminogen-independent fibrinolytic activity, presumably reflecting access of cytoplasmic proteases of broken basophils to fibrin substrate. The PA activity of intact cells was destroyed by gentle trypsinization under conditions that did not impair cell viability. When disrupted cells were ultracentrifuged on a sucrose density gradient, PA activity was absent from purified granules and was confined to fractions containing cell membranes. The simplest explanation of these data is that guinea pig basophils have PA activity associated with their plasma membranes. This conclusion has several important implications for basophil functions in cell-mediated and other immunologic reactions in vivo.
Delineation of Spontaneous Erythrocyte Autoantibody Responses of NZB and Other Strains of Mice
Authors: Deheer David H. , Linder Ewert J. , Edgington Thomas S.
Published: December 2022

Four anti-erythrocyte autoantibody responses (anti-X, anti-HB, anti-HOL, and anti-I) that occur spontaneously in mice have been characterized with regard to antigenic specificities, predominant immunoglobulin class, and pathogenetic importance. Each autoantibody response exhibits specificity for an independent erythrocyte membrane autoantigen (X, HB, HOL, or I) or a soluble analogue (SEA-X or SEA-HB) present in the plasma. The anti-X response, unique to NZB mice, is directed to a normally exposed murine erythrocyte autoantigen, whereas the anti-HB response is directed to a cryptic erythrocyte autoantigen exposed by limited enzymatic cleavage of the membrane. The anti-I response also is directed to a cryptic but distinct autoantigen, and anti-HOL autoantibodies react with an erythrocyte autoantigen located at the cytoplasmic surface of the membrane.

Analysis of the predominant immunoglobulin class of each of the autoantibodies has demonstrated that anti-HB and anti-I antibodies are predominantly of IgM class, whereas anti-X and anti-HOL antibodies are IgG immunoglobulins. Only anti-X and anti-HB autoantibodies are recovered from Coombs' positive erythrocytes from NZB mice and erythrocytes with surface C3 are detected only in NZB mice greater than 9 months of age. These data suggest that only the anti-X and anti-HB responses are pathogenetically implicated in the autoimmune hemolytic anemia of NZB mice.
Leukocyte Complement: Neoantigens of the Membrane Attack Complex on the Surface of Human Leukocytes Prepared from Defibrinated Blood
Authors: Sundsmo John S. , Kolb William P. , Müller-Eberhard Hans J.
Published: December 2022

The neoantigenic determinants (neoAg) which have been identified in the human C5b-9 membranolytic C complex were detected here by the direct fluorescent antibody technique on the surface of 27 ± 11% of viable peripheral blood leukocytes (PBL). The cells were prepared from defibrinated blood by sedimentation on Ficoll-Hypaque. Specificity of the antisera was established by quantitative inhibition of the fluorescent staining reaction, and of agglutination of EAC1-7, by highly purified C5b-9 complex. No inhibition was observed with fresh normal human serum. The majority of the PBL with surface neoAg was found in the B lymphocyte subpopulation that failed to form rosettes with sheep erythrocytes. NeoAg on B lymphocytes was removed to differing degrees by trypsin, papain, or pepsin treatment, and by maintaining the cells at 4°C for 20 hr in serum-free medium. The individual components, C5, C6, C7, C8, and C9, were also detected on the surface of PBL. With differential fluorescent stains, C5 and neoAg as well as C8 and neoAg could be detected on the same cells. The results indicate that viable B lymphocytes prepared from defibrinated blood, have the components of the membrane attack complex of C on their surface. The concomitant occurrence of the neoAg indicates that these proteins are present at least in part in the form of the assembled terminal complex.
Stimulation of the Synthesis and Release of Lipids in Tumor Cells under Attack by Antibody and C
Authors: Schlager Seymour I. , Ohanian Sarkis H. , Borsos Tibor
Published: December 2022

Antibody-sensitized line-1 or line-10 tumor cells treated with GPC (TAC) incorporated fatty acids into complex cellular lipids and released increased amounts of fatty acids within 5 to 10 min after the addition of GPC as compared to control cells. This effect was dependent on the concentration of GPC used; however, under conditions where the cells were not killed, the enhanced synthesis and release of lipids were not dependent on the antibody concentration used to sensitize the cells. Treatment of the cells with antibody alone, GPC alone, or antibody plus heat-inactivated GPC did not result in enhanced synthesis or release of lipids. No enhancement in DNA, RNA, or protein synthesis in TAC was noted. Line-1 cells, which can be killed by GPC when sensitized with excess anti-Forssman IgM antibody, demonstrated enhanced lipid synthesis within 1 to 3 min after the addition of GPC to the antibody-sensitized cells, before measurable killing of the cells had occurred. This effect persisted in the surviving cells when tested 5 and 10 min after the formation of TAC. Addition of GPC deficient in C4 to antibody-sensitized cells did not result in enhanced lipid synthesis or release. These data suggest that the synthesis of macromolecules of which lipids are a major component is of central importance for the ability of the cells to resist antibody-GPC mediated attack.
Control of Human B Lymphocyte Responsiveness: Enhanced Suppressor T Cell Activity after in vitro Incubation
Authors: Lipsky Peter E. , Ginsburg William W. , Finkelman Fred D. , Ziff Morris
Published: December 2022

Human peripheral blood mononuclear cells (PBM) lost the capacity to generate immunoglobulin-secreting cells (ISC) in response to pokeweed mitogen (PWM) after a period of preincubation in vitro. When fresh PBM were co-cultured with preincubated PBM their response to PWM was inhibited, indiating that enhanced suppressor activity developed in the aged PBM concomitant with the loss of PWM responsiveness. Suppressor cell activity of aged PBM was present within the T lymphocyte population. The suppressor T cell inhibited PWM responsiveness of autologous and homologous PBM to an equivalent degree. The action of the suppressor cell was abrogated by inhibitors of DNA synthesis or by hydrocortisone. A suppressor T cell population with similar characteristics was found in freshly prepared PBM before in vitro incubation. Expansion of this suppressor T cell population during preincubation required cell division. There was no change in the functional capability of the helper T cell population as a result of similar in vitro culture. These observations indicate that a T cell population capable of suppressing PWM-induced generation of ISC can be selectively expanded by in vitro incubation of normal human PBM without additional mitogenic stimulation. Moreover, these data emphasize that control of B lymphocyte differentiation involves a critical interrelationship between T lymphocyte subpopulations exerting both positive and negative influences.
Production of Migration Inhibition Factor (MIF) and an Inducer of Plasminogen Activator (IPA) by Subsets of T Cells in MLC
Authors: Newman Walter , Gordon Saimon , Hämmerling Ulrich , Senik Anna , Bloom Barry R.
Published: December 2022

Supernatants of murine spleen cells activated in MLC contain both MIF and a factor which induces plasminogen activator secretion by macrophages. These activities are not found in supernatants of unstimulated T cells in control (syngeneic) cultures. Because no plasminogen activator or fibrinolytic activity was detected in the lymphocyte supernatants directly, but appeared when supernatants were added to macrophages, we infer that production of this activity is induced in macrophages by an inducer of plasminogen activator (IPA).

IPA and MIF activities are first detected after 24 hr of MLC; peak activity occurred on day 3. Both factors eluted from Bio-Gel P-100 columns in the range of Kd 0.15 to 0.25 (m.w. range 25,000 to 60,000 daltons).

Production of both factors in this system required the presence of T cells. In experiments using anti-Ly 1 and anti-Ly 2,3 sera + C, it was observed that both the Ly 1 (helper-type) and Ly 2,3 (cytotoxic/suppressor-type) T cell subpopulations produced each mediator. It is likely that the production of mediators by individual T cell subsets in vivo is not restricted by their intrinsic capabilities, but rather by factors which determine their activation. These results point to the possible importance of both T cell subpopulations in activating macrophages, and indirectly in aspects of tissue injury resulting from cell-mediated immune reactions involving protease activation.

Since IPA correlated with MIF in terms of activity (0.78 coefficient of correlation), kinetics of production, gel filtration characteristics, and production by T cell subsets, and because the assay for plasminogen activator is a sensitive and reproducible one, it should be increaseingly useful in in vitro studies of cell-mediated immunity.
DAN Synthesis and Proliferation of Human Lymphocytes in Vitro
Authors: Bernheim Jan L. , Mendelsohn John
Published: December 2022

DNA newly synthesized by phytohemagglutinin-(PHA) stimulated human lymphocytes has been analyzed for the possibility that all DNA synthesis may not represent premitotic genome duplication. Equilibrium density gradient characterization of bromodeoxyuridine-(BUdR) substituted DNA demonstrates semi-conservative DNA replication, without evidence for repair synthesis. Experiments to detect selective replication (amplification) of a portion of the DNA involved measurement of reassociation kinetics as well as measurement of the kinetics of BUdR appearance in prelabeled DNA. The presence of large quantities of amplified DNA has been excluded, although amplification cannot be ruled out completely by these techniques. Finally, the kinetics of DNA release from dead lymphocytes is characterized, and factors tending to reduce the complexity of released, labeled DNA are identified.
Enhancement of Carrier-Specific Helper T Cell Function by the Synthetic Adjuvant, N-Acetyl Muramyl-l-Alanyl-d-Isoglutamine (MDP)
Authors: Sugimoto Masanobu , Germain Ronald N. , Chedid Louis , Benacerraf Baruj
Published: December 2022

The adjuvant effect of a synthetic peptidoglycan, muramyl dipeptide (N-acetyl muramyl-l-alanyl-d-isoglutamine, MDP), was studied by using the anti-Tnp PFC and hemagglutinin responses of BALB/c mice to hapten-carrier conjugates. Administration of Tnp-OVA and MDP in saline to mice, followed 2 weeks later by a boost of Tnp-OVA in saline, led to significantly higher IgM and IgG anti-Tnp PFC and total anti-Tnp-hemagglutinin responses than those obtained in mice not treated with MDP in the initial immunization. A similar adjuvant effect by MDP on anti-hapten PFC responses was seen if mice were primed with KLH together with MDP and challenged with Tnp-KLH 2 weeks later. This apparent effect on carrier priming for helper function was confirmed and quantitated by double adoptive transfer experiments with graded numbers of spleen cells from KLH ± MDP-primed mice and a fixed number of haptenprimed spleen cells from syngeneic Tnp-OVA immunized animals. These data suggest that at least one mode of action of the synthetic adjuvant MDP is via the enhanced stimulation of the helper T cell function.
Interpretation of 51Cr-Release Data: A Kinetic Analysis
Authors: Thoma John A. , Touton Mary H. , Clark William R.
Published: December 2022

An analysis of 51Cr-release data from cell-mediated cytotoxicity assays, plotted in the form of effector cell dilution curves, is analyzed in detail in terms of a saturation kinetics model for effector-target interaction. The effect of nonimmune (bystander) cells in the effector population is particularly examined. The theoretical development of the saturation model predicts that, at low concentrations of target cells, increasing proportions of bystander cells should broaden the range of effector cell concentration required to achieve a given increment of target cell lysis, but that at high target cell concentration this effect should disappear. Experimental validation of this prediction is presented.
Ionophore A 23187-Induced Production of Slow Reacting Substance of Anaphylaxis (SRS-A) by Rat Peritoneal Cells in Vitro: Evidence for Production by Mononuclear Cells
Authors: Bach Michael K. , Brashler John R.
Published: December 2022

A slow reacting substance is produced when mixed peritoneal cells from rats are incubated with low concentrations of the calcium ionophore A 23187 in the presence of 0.01 M cysteine. The product resembles slow reacting substance of anaphylaxis (SRS-A), if it is not identical to it, based on its behavior during chromatography on Amberlite XAD-2, silicic acid, and Sephadex LH-20; its destruction by dilute HCl and its stability in dilute alkali; its susceptibility to inactivation by aryl sulfatase; and its smooth muscle contracting activity on the guinea pig ileum and the gerbil colon. In vivo selective depletion procedures and in vitro cell isolation procedures were employed to identify the cells responsible for SRS-A production. Mast cells and mixtures of neutrophils and eosinophils produced only very small amounts of SRS-A; this could not be explained by the destruction of preformed SRS-A by these cells. The low yield of SRS-A from mast cells was also not explained by feedback inhibition caused by the histamine they released, because high concentrations of the H2 antagonist, Burimamide, caused only modest increases in SRS-A production by mast cell-containing cell suspensions. Granulocyte-depleted cell suspensions which were derived from thioglycolate-induced mononuclear cell exudates produced more SRS-A per cell than did comparable preparations from uninduced rats. The SRS-A production by induced monocytes was virtually abolished in tandem with the depletion of the phagocytic cells in the preparation. The possible implications of these findings on the source and role of SRS-A in the rat are briefly considered.
Cytotoxic Activities of Normal Cultured Human T Cells
Authors: Alvarez Jose M. , de Landazuri Manuel O. , Bonnard Guy D. , Herberman Ronald B.
Published: December 2022

Normal human T cells grown in continued cultures in medium containing conditioned medium (CM) from PHA-stimulated lymphocytes were studied for their ability to manifest three known forms of cell-mediated cytotoxicity: lectin-induced cellular cytotoxicity (LICC), natural killer cell (NK) activity, and antibody-dependent cellular cytotoxicity (ADCC). The cultured T cells (CTC) were very effective mediators of LICC, being cytotoxic even at very low attacker-target cell ratios in the presence of different lectins, and against different types of targets. When tested without the addition of lectin, the CTC demonstrated a low degree of spontaneous cytotoxicity. This spontaneous cytotoxicity might not be due to conventional NK cells however, since the CTC failed to show significant numbers of cells with Fc receptors (FcR) for IgG, and had no detectable ADCC activity. CTC could represent a population enriched in polyclonal activated T cells with low spontaneous cytotoxicity against a variety of allogeneic target cells, which is greatly enhanced by the addition of lectins during the 51Cr release assay.
Antigen-Specific T Cell-Mediated Suppression
Authors: Pierres Michel , Germain Ronald N.
Published: December 2022

The role of adherent cells (MØ) in 1) primary in vitro PFC responses and 2) in vitro generation of suppressor T cells (Ts) to the antigen L-glutamic acid60-L-alanine30-L-tyrosine10 in responder strain mice has been investigated. Removal of MØ from spleen cell suspensions by either serial culture on plastic surfaces or passage over Sephadex G-10 beads yielded lymphocytes unable to give primary in vitro PFC responses under modified Mishell-Dutton conditions to either soluble GAT or the particulate antigens sheep red blood cells (SRBC) or GAT-complexed electrostatically to methylated bovine serum albumin (GAT-MBSA). However, the viability of such MØ-depleted cultures was significantly lower than control cultures of unseparated spleen cells. Addition of 5 × 10-5 M 2-mercaptoethanol (2-ME) to the culture medium restored viability and permitted nonadherent cells to respond to SRBC and GAT-MBSA, but not to soluble GAT. The response of nonadherent cells to soluble GAT could be restored in the absence of 2-ME by the addition of 1 to 3 × 105 x-irradiated, anti-Thy 1.2 + C-treated peritoneal exudate cells (MØ), or in the presence of 2-ME by 1 to 2 × 104 MØ. This low number of MØ was identical to the number of GAT-bearing MØ necessary in the presence of 2-ME to give optimal GAT PFC responses. Taken together, these data indicate a crucial antigen-presenting role for MØ in primary in vitro anti-GAT responses. Removal of such antigen-presenting cells from responder spleen cells left a lymphocyte population, which, when cultured for 2 days with normally immunogenic (10 µg/ml) concentrations of GAT, gave rise to highly active Thy 1.2+ GAT-specific suppressor cells. This Ts induction was demonstrated in several responder strains. The GAT-Ts were unable to suppress the in vitro responses to SRBC or the copolymer L-glutamic acid50-l-tyrosine50 (GT) as GT-MBSA. Increasing the concentration of GAT to ≥ 30 µg/ml permitted the induction of GAT-Ts in unseparated spleen cells. Thus, in responder mice, MØ able to present antigen in an immunogenic form play a central role in regulating the balance of activated TH and Ts, suggesting that an Ir gene defect in nonresponder mice at the MØ level for this presentation function may account for the predominant suppressor T cell responses of such mice.
Immunoglobulin M Synthesized by Human Lymphoblastoid Cells: Interaction with Staphylococcus Aureus and Protein A
Authors: Howell-Saxton Ernestina , Wettstein Felix O.
Published: December 2022

Immunoglobulin M synthesized by a human lymphoblastoid cell line, LA173, was found to bind specifically to the protein A-bearing Cowan I strain of Staphylococus aureus. The (3H)-leucine-labeled, secreted IgM from these LA173 cells also formed precipitin complexes with purified protein A. Soluble complexes formed at high protein A/IgM ratios retained the ability to bind to the bacterial surface. Precipitin complexes also were observed in double diffusion Ouchterlony gels with a line of identity formed between the IgM, protein A, and anti-IgM in adjacent wells.

Intracellular IgM species from detergent-lysed LA173 cells were bound to S. aureus. Labeled 19S pentamers, 8S monomers, and HL subunits were eluted from the bacteria and identified by velocity sedimentation and SDS agarose-acrylamide gel electrophoresis. In addition, several intermediates migrating between 8S and 19S were detected and shown to contain authentic H and L chains. Binding of the labeled IgM 19S pentamers to staphylococci was not inhibited by prior treatment of the bacteria with an excess of unlabeled human IgG. However, S. aureus saturated with unlabeled IgG did not bind either labeled IgM monomers or labeled IgG. The interaction of this human IgM with S. aureus exhibited a high degree of specificity with quantitative recovery of secreted 19S IgM. Intracellular IgM species were bound selectively by the bacteria with little if any contamination by other cytoplasmic proteins.
Expression of Human T Lymphocyte Antigens by Killer Cells
Authors: Kaplan Joseph , Callewaert Denis M. , Peterson Ward D.
Published: December 2022

K cells, the effectors of antibody-dependent cell-mediated cytotoxicity, were found to express human T but not B lymphocyte antigens detected by rabbit anti-HTLA and anti-HBLA. Pretreatment of effector cells with anti-HTLA+C inhibited ADCC by specifically lysing K cells: no inhibition of ADCC by anti-HTLA occurred when ΔC was substituted for C. By contrast, pretreatment of effector cells with anti-HBLA nonspecifically inhibited ADCC, probably by forming antigen-antibody complexes with HBLA+ cells in effector suspensions: a) treatment with anti-HBLA+ΔC was more inhibitory of ADCC than treatment with anti-HBLA+C, and b) the inhibitory effect of anti-HBLA on ADCC was either eliminated or markedly reduced if effector suspensions were first passed through a nylon fiber column, a procedure that removed most HBLA+ cells without affecting K cell activity. HTLA antigens expressed by K cells and NK cells are the same as HTLA antigens expressed by thymocytes since thymocytes completely absorb the anti-K cell and NK cell reactivity of anti-HTLA.
Induction of B Cell Priming by Neonatal Injection of Mice with Thymic-Independent (Type 2) Antigens
Authors: Mosier D. E.
Published: December 2022

Mice injected at birth with the thymus-independent type 2 antigen TNP-AECM-Ficoll have augmented anti-TNP antibody responses when their spleen cells subsequently are challenged in vitro with TNP-coupled thymic independent or thymic dependent antigens. This neonatal priming effect was shown to occur in neonatal nu/nu mice and thus does not appear to require T lymphocytes. The primary explanation for the priming effect seems to be an increase of approximately 10-fold in the numbers of TNP-specific precursors of antibody-forming cells. The neonatal injection of TNP-AECM-Ficoll induces little or no antibody formation directly. It appears, therefore, that some thymic independent antigens can deliver a signal to immature B cells, which causes clonal expansion, but is unable to induce differentiation into antibody-forming cells.
Effect of Splenectomy on the Expression of Regulatory T Cell Activity
Authors: Amsbaugh Diana F. , Prescott Benjamin , Baker Phillip J.
Published: December 2022

The effect of adult splenectomy on the expression of suppressor and amplifier T cell activity was examined with respect to the serum antibody response to Type III pneumococcal polysaccharide (SSS-III) by using a sensitive radioimmunoassay. Suppressor T cell activity, as measured by the degree of low-dose paralysis induced, was not impaired in the least by splenectomy; however, amplifier T cell activity was almost completely eliminated within 7 days after splenectomy. These findings indicate that suppressor T cell activity is not confined solely to the spleen, the major site of antibody synthesis after immunization with SSS-III, and that the spleen may be an important site for the generation and/or maintenance of amplifier T cell activity.
Double Expression of VH Region a2 and a3 Allotypes in Individual Rabbit Lymphocytes
Authors: Wolf Benjamin , Wang Shaw-Shyan
Published: December 2022

This study provides evidence that two VH region a group allotypic specificities are synthesized by individual peripheral blood lymphocytes from a2,3 rabbits. After removal of both all types from the cell surface with pronase, three types of allotype-bearing cells, a2, a3, and a2,3 were demonstrable after overnight culture in serum-free medium. Frequently, greater than 50% of the cells that contained allotype on the surface resynthesized both allotypes. In culture, reappearance of allotype was first detectable at 4 hr and increased progressively thereafter to a maximum at 18 hr. During regrowth, the relative proportion of cells showing both markers was maintained. The predominance of cells exhibiting only a3 over those displaying only a2 was also maintained. Reappearance of allotype was inhibited by cycloheximide but was manifested when the antibiotic was washed out. Monospecific anti-a2 antibodies could completely suppress the resynthesis of homologous allotype in culture and prevent the regeneration of a2 in double-bearing cells. Likewise, anti-a3 in cultures was usually specifically suppressive for a3 allotype; however, the double producers were only partially depleted.
Regulation of Delayed-Type Hypersensitivity Reactions by Cyclophosphamide-Sensitive T Cells
Authors: Schwartz Anthony , Askenase Philip W. , Gershon Richard K.
Published: December 2022

The onset, intensity, and duration of DTH reactions elicited in mice immunized with either SRBC or products of the major histocompatibility complex can be altered significantly by pretreatment with CY 1 to 2 days before immunization. Such drug pretreatment tends to augment low DTH responses caused by the use of too much antigen and to diminish many responses that are optimal. Thus, pretreatment with CY does not specifically eliminate suppressor cells. Our results are most consistent with the notion that the cellular targets of low doses of CY are positive and negative feedback regulatory cells, which may consist of one population with two effects or, more likely, two distinct cell populations.
Quantitative Differences in Immune Responses during the Various Stages of the Estrous Cycle in Female BALB/c Mice
Authors: Krzych Urszula , Strausser Helen R. , Bressler Joseph P. , Goldstein Allan L.
Published: December 2022

The immune responsiveness of spleens from female BALB/c mice to PHA, Con A, and LPS was greater at proestrus and metestrus as compared with estrus and diestrus. The peaks of responsiveness corresponded to reported elevated levels of estrogen and pregnenolone during these phases of the cycle. Similar results were obtained with the IgM or direct plaque-forming cell responses, which were also increased at proestrus and metestrus. It appears that female hormones may directly or indirectly stimulate immune responsiveness in adult mice.
Monoclonal Antibodies to H-2 and Ia Antigens
Authors: Oi V. T. , Goding J. W. , Jones P. P. , Herzenberg L. A. , Herzenberg L. A.
Published: December 2022

Seven stable hybrid cell lines producing monoclonal IgG antibodies reacting with H-2 and Ia antigens have been established. The lines were obtained by fusing NS-1, a nonsecreting variant of MOPC-21 (Köhler, G., Howe, S. C., and Milstein, C., Eur. J. Immunol. 1976. 6: 292) with spleen cells from either BALB/c or C3H.SW.Igb (CWB) mice that had been immunized with H-2k haplotype spleen cells. One of the monoclonal antibodies reacts with H-2Kk determinants; the remaining six react with determinants mapping to the I-Ak subregion. The antigens bound by the anti-I-Ak antibodies were analyzed by two-dimensional electrophoresis and were found to consist of the same polypeptide chains as are precipitated by anti-I-Ak alloantisera. The Ia antigenic determinants recognized by these monoclonal antibodies and their presence on lymphocyte subpopulations have been characterized by 125I-binding assays, complement-mediated cytotoxicity, and immunofluorescence, by using the fluorescence-activated cell sorter (FACS).
Functional Differences Between Macrophage Subpopulations
Authors: Cowing Carol , Pincus Seth H. , Schwartz Benjamin D. , Sachs David H. , Dickler Howard B.
Published: December 2022

The expression of Ia antigens on murine macrophages was studied by indirect immunofluorescence, microcytotoxicity, and biosynthetic labeling. The majority of peritoneal macrophages lacked Ia antigens. When splenic macrophages were examined, it was found that the majority of these cells expressed Ia antigens and that these were synthesized by the macrophages. All the splenic macrophages which reacted with alloantibody raised against the whole I region expressed I-A subregion antigens, while only a subset of these cells also expressed I-E or C subregion antigens.

When antigen-pulsed normal peritoneal and spleen macrophage-enriched cell populations were compared for their ability to stimulate antigen specific proliferation of primed lymph node T cells, a positive correlation was found between the number of Ia bearing phagocytic cells and T cell DNA synthesis. Spleen adherent cells and T cells were then treated with subregion restricted anti-Ia antisera and complement.
Three Separable Molecules Bearing Ia Antigens Determined by the H-2d Haplotype
Authors: Sachs David H. , Hansen Ted H.
Published: December 2022

A (B10 × D2.GD)F1 anti-B10.D2 antiserum was prepared in order to examine Ia antigens determined by genes to the right of the I-A subregion of the H-2d haplotype. This antiserum complement-mediated lysis of H-2a and H-2d spleen cells to a titer of 1:256, indicative of its activity against Dd antigens. Absorption with B10.A spleen cells left anti-Ia (B cell) cytotoxic reactivity to a titer of 1:64 when tested on H-2d spleen cells. This correlates with, and helps to define, the recently described I-E subregion specificity, Ia.23, of the H-2d haplotype (C. S. David, in press). Absorption of this antiserum with B10.BR spleen cells remove all cytotoxic activity against B10.A(2R) spleen cells, indicating either that the serum contained no detectable antibodies to private Ia antigens determined by the I-Cd subregion or that the I-C subregion of H-2a is not identical to that of H-2d.
Structural Analysis of Murine and Human Ia Alloantigens
Authors: Allison J. P. , Walker L. E. , Russell W. A. , Ferrone S. , Reisfeld R. A. , Pellegrino M. A. , Frelinger J. A. , Silver J.
Published: December 2022

We are determining the structure of the murine Ia alloantigens determined by the IA and IC subregions of the major histocompatibility complex (MHC) and comparing their structure to their human counterparts, i.e, DR antigens. Murine Ia and human DR antigens are isolated by indirect immunoprecipitation using highly specific allo and xenogeneic antisera, respectively. The molecular properties and N-terminal sequence of these cell surface proteins is determined using procedures that have previously provided detailed structural information for the K and D gene products of the MHC. Ia antigens determined by the IA and IC subregions each consist of two non-covalently associated molecules of 35,000–37,000 daltons and 28,000–29,000 daltons while the human DR alloantigens consist of two noncovalently associated molecules of 34,000 daltons and 29,000 daltons. Partial N-terminal sequence analysis indicates that while there is no apparent homology between the murine Ia alloantigens determined by the IA and IC subregions, the human DR alloantigen is structurally equivalent to the IC subregion alloantigen.
Structure of Guinea Pig Ia Antigens
Authors: Waxdal M. J. , Basham T. Y. , Clement L. , Shevach E. , Schwartz B. , Tyran B.
Published: December 2022

Guinea pig Ia molecules bearing the markers 3, 5 and 7 (strain 13) and 4, 5 (strain 2) were isolated from spleen and lymph node cells. In the molecules bearing the 7 and the 4, 5 markers, the 25,000 and 33,000 MW polypeptide chains are linked by disulfide bonds. The molecules bearing the 3, 5 markers do not have polypeptide chains linked by disulfide bonds. These glycoproteins were metabolically labeled with tritiated amino acids and purified by NP-40 lysis, affinity chromatography on columns of lentil lectin and specific immunoabsorbents, followed by SDS-PAGE. In some cases the immunoabsorbent chromatography was replaced by immunoprecipitation using SaCI. The radioactive polypeptide chains were eluted from the gel and characterized by peptide mapping and by sequence analysis of the N-terminal regions of the 25,000 MW chains. The preliminary results indicate that all three 25,000 MW chains studied were homologous.
Influence of Non H-2 Linked Ir Genes on the Response of Mice to Insulins of Different Species
Authors: Keck Klaus , Momayezi Massoud
Published: December 2022

We have previously shown that the IgG response of mice to different insulins is controlled by H-2 linked Ir genes. Furthermore, the A-Chain loops of some insulins act as carrier determinants. The response to insulins is inherited as a dominant or recessive trait. More recent experiments show that in addition to H-2 linked Ir genes, other genes play an important role in the response to these antigens. (a) Complementation of two nonresponding strains to a high responding hybrid can only be achieved if at least one of the parental strains has a C3H background. (b) Induction of tolerance to the immunogenic sheep insulin can be induced by preimmunisation with the less immunogenic pig insulin only in C3H/He mice, but not other strains bearing the same H-2 haplotype. (c) A primary IgM response to pig insulin can only be induced in B10. BR but not in C3H/He mice, although both strains carry the same H-2 haplotype.
Determinant Selection by Mouse Strains Responding to (Glu Lys Phe)Terpolymers: Relevance to the Ir Gene Complementation Phenomenon
Authors: Maurer Paul H. , Merryman Carmen F. , Lai Chang-Hai , Babu Uma Mahesh
Published: December 2022

Based upon our findings that copolymers of (Glu Phe) are immunogenic in mice of H-2 haplotypes a, k, p and q, and experiments done in collaboration with Drs. Baltz and Feldmann which indicated that H-2k mice respond to GLPhe11, we have reinvestigated the immune responses of the inbred, congenic and recombinant mice to GLPhe terpolymers. In addition to analyzing responses for antibody, the in vitro T cell proliferative responses were assayed with a number of “cross-reacting” immunogenic and nonimmunogenic terpolymers. Several important findings were noted: 1) Antibody against GLPhe was produced by mice of H-2 haplotypes a, d, k, p and q. 2) H-2d T cells responded to GLPhe11, GLPhe5 and GLT5, but not to GPhe. 3) H-2k T cells were stimulated by GLPhe11, weakly by GPhe, but not by GLPhe5 or GLT5. 4) H-2p T cells were stimulated by GLPhe11, weakly by GLPhe5 and GPhe, but not by GLT5.
Ir Gene Control of T Cell Responses to the Male Specific Antigen H-Y
Authors: Simpson Elizabeth , Hurme Mikko
Published: December 2022

Female mice of the H-2b haplotype possess a dominant Ir gene controlling H-2 restricted cytotoxic T cell responses to H-Y. This gene maps in the IA subregion, but requires appropriate associative antigens coded by K or D end genes in order for the response to be initiated and effected. Separable from the Ir gene controlling anti-H-Y cytotoxic responses in H-2b mice is a dominant Ir gene in the IB subregion and this controls the ability of female mice to reject syngeneic male skin grafts. It does not apparently require associative K/D antigens for its operation.

Mouse strains of independent haplotypes other than H-2b are nonresponders to H-Y, both with respect to the generation of H-2 restricted cytotoxic responses and to skin graft rejection. However, certain F1 hybrids (e.g. k/d, k/s, k/a, k/q, d/s) are responders in that they make cytotoxic responses to H-Y, although none of them reject skin grafts.
Functional Capacities of Ia-Bearing Langerhans Cells
Authors: Stingl G. , Katz S. I. , Clement L. , Green I. , Shevach E. M.
Published: December 2022

Langerhans cells (LC) represent a small subpopulation of the mammalian epidermis. Though morphologically well characterized the functional role of these cells is still unknown. The previous hypothesis that LC are functionally related to macrophages is strongly supported by the recent finding that human and guinea pig epidermal LC express Fc-IgG receptors, C3 receptors and Ia antigens. In this study we investigated whether guinea pig LC can mediate the same immunologic functions as Ia-bearing macrophages. LC-enriched (LC-e) and LC-depleted (LC-d) epidermal cells were prepared by separation of Fc-IgG rosetting epidermal cells on density gradients. The percentage of LC ranged from 18–35% in the enriched fraction and from 0.2–0.5% in the depleted fraction as evaluated by histochemical staining techniques. Virtually all the Ia activity was restricted to the LC-e population as determined by immunoprecipitation experiments.
Role of Suppressor T Cells in Ir Gene Restricted Macrophage-T Cell Interactions in (Responder × Nonresponder)F1 Mice Primed to l-Glutamic Acid60-l-Alanine30-l-Tyrosine10 (GAT)
Authors: Germain Ronald N. , Benacerraf Baruj
Published: December 2022

(Responder (R) × Nonresponder (NR))F1 mice give indistinguishable primary PFC responses in vitro to GAT-pulsed parental R or NR macrophages (Mϕ). It was recently shown that such (R × NR)F1 mice, if primed to GAT, retained responsiveness to GAT-R-Mϕ, but no longer responded to GAT-NR-Mϕ, suggesting a possible Mϕ-related locus for Ir gene activity in this model. We have now found that Ir related restriction in such GAT-primed F1 mice is at least in part related to the stimulation of suppressor T cells. (B6 × D1)F1 (H-2b/q) mice were primed with 100 µg GAT in Maalox with pertussis. Some mice were pretreated with 5–10 mg/kg cyclophosphamide (CY) 3 days prior to immunization. Two to eight weeks later, the spleens of these mice were tested in vitro for PFC responses to GAT-pulsed parental Mϕ. Primed F1 mice responded only to GAT-B6-Mϕ. CY pretreated mice responded to both GAT-B6-Mϕ and GAT-D1-Mϕ.
I-J Region Regulation of Macrophage-T Lymphocyte Interaction in the in Vitro Antibody Response
Authors: Niederhuber John E. , Mayo L. , Allen Paul
Published: December 2022

The specificity of macrophages (mϕ) in regulation of the antibody response is important to the understanding of antigen processing, Ir-gene control, and mϕ-T cell interaction. We have demonstrated the requirement for Ia bearing macrophages in the in vitro antibody response. Using restricted anti-Ia sera, experiments were designed to determine the critical I subregion for this region. 107 mϕ depleted T and B spleen cells were added to 5–10 × 105 anti-Thy 1.2 plus anti-Ig sera purified mϕ. The mϕ fraction was then treated with either anti-Ia sera plus rabbit complement, thus deleting Ia+ mϕ, or they were treated only with the Ia antisera, which blocked appropriate Ia surface molecules. Both methods were used in the same experiment. Alloantisera specific for the I-J subregion with or without complement were equally capable of significant inhibition of the plaque forming response. Anti-Ia sera specific for products of other I subregions, for example the I-A subregion, eliminate mϕ function only in the presence of complement.
H-2 Linked Responsiveness Is Acquired by Low Responder Lymphocytes Differentiating in the Thymus of High Responders
Authors: von Boehmer H. , Haas W.
Published: December 2022

Cytotoxic T cells appear to recognize Y-antigen and H-2 as distinct entities: 1) XX H-2b T cells depleted of cells reactive to H-2k alloantigens fail to recognize putative neoantigens formed by H-2k and Y-antigens while reacting to XY H-2b cells. 2) XX H-2b T cells from XX H-2b → XX (H-2k × (H-2b) chimeras can be induced to lyse XY H-2b as well as XY H-2k targets. In contrast XY H-2b T cells from such chimeras lyse neither XY H-2b nor XY H-2k targets.

H-2i5 and H-2k mice are low responders to Y-antigen. H-2i5 lack Y-specific precursors for killer cells while H-2k mice lack Y-specific helpers supporting expansion and/or differentiation of Y-specific killers: H-2i5 lymphocytes differentiating in (H-2k × H-2b) X-irradiated recipients acquire responsiveness to Y-antigen while H-2k lymphocytes do not. (H-2k × H-2b) lymphocytes differentiating in H-2k recipients lose the ability to respond to Y-antigen.
Adaptive Differentiation of Murine B and T Lymphocytes
Authors: Katz David H. , Skidmore Barry J.
Published: December 2022

The concept of adaptive differentiation predicts that early differentiation of lymphocytes can be conditioned by the environment in which such differentiation takes place such that the capacity for most effective cell-cell interactions between such lymphocytes, which is controlled by major histocompatibility complex-linked genes, is learned in the process of such differentiation. In these studies, we have tested the capacities of both helper T lymphocytes and hapten-primed B lymphocytes primed in the environments of all combinations of bone marrow chimeras prepared between two parental strains (i.e. A/J and BALB/c) and their corresponding F1 hybrid (CAF1) to interact with cooperating B or T cells derived from conventional parental and F1 donors as well as all of the corresponding bone marrow chimera combinations.
In Vitro Studies on the Cellular Site of Ir-Gene Expression
Authors: Kappler John W. , Marrack Philippa
Published: December 2022

(TG)-A--L-primed T cells from high responder × low responder F1 mice induced high anti-TNP responses to TNP-(TG)-A--L in vitro in B cells and macrophages (Mϕ) from the high responder parent, but low responses in B cells and Mϕ from the low responder parent. Mixing experiments established that high responsiveness required the presence of both high responder B cells and Mϕ in the cultures demonstrating the expression of Ir-genes in both of these cell types.

Irradiation chimeras were prepared by transferring parental bone marrow into irradiated F1 recipients. (TG)-A--L-primed T cells bearing the low responder H-2 type were isolated from these chimeras and shown to induce high responses in vitro with high responder B cells and Mϕ. Thus, it was not obligatory that helper T cells carry the high responder Ir-gene in order to obtain a high response.

Irradiation chimeras were prepared by transferring F1 bone marrow into irradiated low responder parental mice.
Genetic Control of Immune Suppression
Authors: Kontiainen Sirkka , Simpson Elizabeth , Feldmann Marc
Published: December 2022

Suppressor T cells induced in vitro by high concentrations of antigen release suppressor factors into their medium if incubated for 24 hours with antigen. Since there are reports that extracts of suppressor cells are related to immune response gene function, the capacity of responder or nonresponder strains to produce suppressor factor (SF) was investigated. Both responder and nonresponder strains produced SF to (T,G)-A--L and GAT. This together with differences in the target and the lack of H-2 restrictions indicates that SF is quite distinct from suppressor cell extracts, despite both having I-J controlled determinants and an antigen combining site.

The structure of SF was analysed using serological means. Rabbit and mouse antisera were raised against SF purified by absorption and elution from antigen columns. Rabbit antisera were not specific for strain or antigen specificity of SF, but did not react with helper factor, and thus defined a “constant region.”