bluuebunny/crossref_metadata_2025 huggingface dataset:
The Frequency of p-Azophenylarsonate and Dimethylaminonapthalene-Sulfonyl-Specific B Cells in Neonatal and Adult BALB/c Mice
Authors: Sigal Nolan H.
Published: December 2022

The limiting cell dilution splenic focus technique has been used to determine the frequency of p-azophenylarsonate (Ars) and 5-dimethylaminonaphthalene-sulfonyl (Dansyl)-specific B cells in neonatal and adult mice. The adult frequencies for Ars and Dansyl in conventionally reared BALB/c mice are 1.5 ± 0.3/105 B cells and 9.4 ± 3.3/105 B cells, respectively. Dansyl-specific precursor cells are present in high frequencies at birth and reach adult precursor frequency levels during the first week of life. This finding mirrors the ontogenetic development previously described for dinitrophenyl (DNP) and trinitrophenyl (TNP), but contrasts with the results obtained for fluorescein (FL) and phosphorylcholine (PC), for which precursors arise later in development. On the other hand, the kinetics of appearance of B cells specific for Ars is unique in that the precursor frequency appears to decrease during the first week of life. The data presented remain consistent with a highly ordered, rigorously predetermined acquisition of the specificity repertoire during ontogeny. By increasing the flexibility of the splenic focus technique to allow analysis of other, chemically diverse haptens, results have been obtained that confirm and extend observations made with DNP, TNP, FL, and PC.
BCG-Induced Murine Effector Cells
Authors: Wolfe Seth A. , Tracey Daniel E. , Henney Christopher S.
Published: December 2022

The cytotoxic activity in peritoneal exudates harvested from C57BL/6 mice 4 to 6 days after they had received viable Bacillus Calmette-Guérin (BCG) organisms i.p. was associated with a nonadherent, nonphagocytic cell. The cytotoxic cell lacked demonstrable surface immunoglobulin and Thy 1 antigen and bore no readily detectable Fc receptors. Lytic activity was labile at 37°C and was diminished after trypsinization of the effector cells. Preincubating effector cells with immune complexes was without effect on lytic expression.

These features make it likely that the cytolytic activity was associated with “natural killer” (NK) cells, previously described in unimmunized mouse spleens and mesenteric lymph nodes. Whether BCG induced “activation” of resident NK cells, the de novo production of NK cells, or “homing” into the peritoneum of cells normally resident in other lymphoid tissue is not known.
Differential Tumor Immunogenicity of L1210 and its Sublines
Authors: Fuji H. , Mihich E. , Pressman D.
Published: December 2022

The differential immunogenicity of DBA/2 lymphoma L1210 and three L1210 sublines, each resistant to a different anti-leukemic agent (guanazole, methylglyoxal-bis-guanylhydrazone, and 4,4-diacetyldiphenylureabis-guanylhydrazone), was evaluated in vitro. Syngeneic spleen cells from nonimmunized DBA/2 mice were cultured in the presence of graded numbers of irradiated cells of L1210 or its sublines. The stimulation elicited a T-independent primary antibody response in vitro which was measured by determining the number of plaque-forming cells by using the immunizing lymphoma cells as target. Cells of all three sublines exhibited an increased immunogenicity, as compared to that of the parental L1210 cells, in eliciting the response directed to tumor-associated antigens which were common to all sublines. Dose-response experiments showed that high doses of the parental cells did stimulate responses which were detectable with subline cells as target. The results indicated that the differential immunogenicity of L1210 and its sublines, as demonstrated in the present assay system, is primarily quantitative, and was apparently due to increased amount or density of common tumor-associated antigens on the subline cells. The implications of these observations are discussed in relation to the possible mechanisms underlying the emergence of highly immunogenic drug-resistant sublines.
Cell-Mediated Immune Response to Lymphocytic Choriomeningitis and Vaccinia Virus in Rats
Authors: Zinkernagel Rolf M. , Althage Alana , Jensen Fred C.
Published: December 2022

The parameters of cell-mediated immune responses of rats to infection with lymphocytic choriomeningitis virus or vaccinia virus were assessed by measuring primary footpad swelling, increased weights of the local lymph nodes, increased numbers of lymphocytes per lymph node, and the course of virus-specific cytolytic activity by these lymphocytes. Except for lack of a defined swelling caused by vaccinia virus injected into the hind footpads of rats, the kinetics of all these responses correlated and were in accord with the usual time course of cellular immune responses. Starting 3 days after infection, peaking at 5 to 7 days, and disappearing after 10 to 12 days, the responses by rats to both viruses were comparable to those found in mice. The phagocytes of these infected rats inhibited the growth of Listeria monocytogenes in vivo, indicating activation of the macrophages by virus-specific cellular immunity. The rat cytotoxic lymphocytes were thymus derived as judged by various criteria: inactivation by an absorbed rabbit anti-rat brain antiserum plus C, susceptibility to anti Thy 1.1 plus C, restriction of the lytic activity within inbred strains and probably by the Ag-B locus, and the kinetics of the response. The cytotoxic T lymphocytes were virus specific since they killed only target cells infected with the same virus but not uninfected cells, or targets that were infected with an unrelated virus.
Kinetics of Antibody and Immunoglobulin-Producing Cells Appearing in Popliteal Lymph Nodes of Mice Stimulated with Horseradish Peroxidase
Authors: Ternynck Thérèse , Rodrigot Maryvonne , Avrameas Stratis
Published: December 2022

Mice were given injections in their hind footpads of peroxidase emulsified in complete Freund's adjuvant. Kinetics of cells producing antibodies and cells producing immunoglobulins without detectable antibody function were monitored in the draining popliteal lymph nodes by using enzyme-immunocytochemical procedures, a local hemolysis plaque assay, and immune cytoadherence tests. These last two techniques were performed with sheep red blood cells (SRBC) coated with either peroxidase or anti-mouse Ig antibody.

The kinetics observed with the immunocytochemical and plaque assay procedures were comparable although at each interval tested a higher number of positive cells was scored with the first procedure. With these two techniques, the following observations were made: 1) After antigen administration, the first detectable antibody- (Ab+) producing cells appeared on day 7 or 8. They were preceded 4 days earlier by the appearance of cells producing immunoglobulins without detectable antibody function (Ig+Ab-). The peak response of Ab+-producing cells occurred on day 14, 4 days after the maximum response of Ig+Ab--producing cells. 2) The vast majority of these Ig+Ab--producing cells was related to the injection of peroxidase and not to the complete Freund's adjuvant. Ig+Ab--secreting cells were found not to represent cells secreting antibodies directed against SRBC, DNP determinants, or bromelain-treated mouse red cells. 3) After a second injection, Ab+ and Ig+Ab--producing cells appeared rapidly and simultaneously in approximately equal numbers; the peak number of Ab+-producing cells was seen on days 5 to 7 whereas the Ig+Ab--producing cells disappeared 3 days after challenge. 4) The ratio of secreting versus synthetizing cells remained constant throughout the response, around 2:10 for Ab+ cell population and 5:10 for Ig+Ab- cell population.

With the immune cytoadherence test, cells forming rosettes with peroxidase-coated SRBC were not found in lymph nodes of unstimulated mice although they appeared 4 days after antigen injection. Their number increased exponentially until day 8 and then more slowly until day 14. Although after a secondary injection the number of rosette-forming cells was increased, the peak value attained only 20% of the peak value observed during the primary response.

The above results indicate that after injection of mice with peroxidase in complete Freund's adjuvant, the following sequence occurs: a) appearance of an immunocyte population bearing peroxidase receptors, b) appearance of an immunocyte population either synthesizing and secreting, or not, immunoglobulins without detectable antibody function, d) appearance of an immunocyte population either synthetizing and secreting, or not, antibody.
Studies on the Mechanism of Antibody-Dependent Polymorphonuclear Leukocyte-Mediated Cytotoxicity
Authors: Clark Robert A. , Klebanoff Seymour J.
Published: December 2022

Human polymorphonuclear leukocytes (PMN) were cytotoxic to mammalian tumor cells (LSTRA) in the presence of anti-LSTRA serum as determined by a 51Cr release assay. Cytotoxicity was observed with effector to target cell ratios as low as 0.25:1 and with incubation periods as short as 30 min. Under the conditions employed, human blood mononuclear cells had only minimal cytotoxic activity. Immunologic specificity was demonstrated by 1) the loss of activity when immune serum was preabsorbed with target cells or replaced with normal (preimmune) serum and 2) the lack of 51Cr release from an unrelated target cell. Viable PMN were required as demonstrated by the loss of cytotoxicity on heating or homogenization. Further, an intact PMN metabolic burst was essential for maximal activity since PMN from patients with chronic granulomatous disease were only minimally active. The myeloperoxidase-H2O2-halide system of the PMN was not responsible for cytotoxicity since activity was normal with myeloperoxidase deficient PMN, was preserved in the presence of azide, cyanide or catalase, and was only slightly dependent on halides. Lack of inhibition by heparin militated against a role for PMN granular cationic proteins. Cytotoxicity was blocked by inhibitors of glycolysis, but not by an inhibitor of oxidative phosphorylation. The use of chelating agents indicated a requirement for divalent actions, primarily Mg++. Inhibition of cytotoxicity by the anti-tubulins, colchicine, and vincristine, suggested a role for microtubules. These studies suggest that antibody-dependent PMN-mediated tumor cell cytotoxicity requires specific antibody, viable PMN, a burst of PMN oxidative metabolism, glycolysis, divalent cations, and microtubular function. The PMN lysosomal constituents, myeloperoxidase and the cationic proteins, did not appear to be involved under the conditions employed.
Effect of Macrophages and Antibodies on in Vivo Growth of Moloney Sarcoma in the Rat
Authors: Miller Glenn A. , Feldman Joseph D.
Published: December 2022

Brown Norway and Lewis rats were challenged with a Brown Norway Moloney sarcoma tumor, MST-1, admixed with nonimmune peritoneal exudate macrophages syngeneic to the host; or admixed with nonimmune peritoneal exudate macrophages and hyperimmune anti-MST-1 antibodies. In vivo growth of MST-1 in BN and Lewis rats was inhibited by admixing Brown Norway or Lewis macrophages, respectively, with BN anti-MST-1 antibodies. The inhibiting BN antibodies were of the IgG2 class, lacking IgG2a antibodies. Brown Norway anti-MST-1 of IgG2 class without macrophages did not affect growth of MST-1. Brown Norway and Lewis anti-MST-1 antibodies of IgG2a class enhanced tumor growth, whether admixed with macrophages or not. Anti-MST-1 antibodies of IgM and IgG1 classes did not influence tumor growth. Peritoneal exudate macrophages removed from Lewis donors 8 to 10 days after inoculation of MST-1 inhibited completely growth of the challenge tumor; macrophages of Brown Norway origin were inhibitory only when harvested from hyperimmune donors, that is, 40 or more days after inoculation of MST-1. Macrophages from hyperimmune donors were specifically cytotoxic to MST-1 and did not inhibit an unrelated syngeneic BN tumor of chemical origin.
Human Lymphocytes Bearing “Ia-Like” Antigens; Absence in Patients with Infantile Agammaglobulinemia
Authors: Hoffman Thomas , Wang Chang Yi , Winchester Robert J. , Ferrarini Manlio , Kunkel Henry G.
Published: December 2022

The new Ia-like antigens, which are primarily expressed on B cells, were studied in detail along with other lymphocyte markers in the peripheral blood of normal individuals and patients with immune deficiency. Rabbit antisera to purified Ia antigens proved of special value in this work although alloantisera gave similar results. The Ia antigens were found on most monocytes and all lymphocytes expressing membrane Ig and in addition on a normal population of cells lacking Ig. Both of these lymphocyte populations were completely absent in patients with infantile agammaglobulinemia with a selective B cell defect. Different findings were obtained for Fc-positive cells. In addition to T cells with Fc receptors, a non-T population was apparent that lacked both surface Ig and Ia determinants; these two cell types were not reduced in patients with infantile agammaglobulinemia.

These findings suggest that all the Ia-positive lymphocytes, both those with and without clearly detectable Ig, belong to the B cell lineage. The special value of the Ia antigens as markers for distinguishing lymphocyte subpopulations was apparent in this study.
Induction of Experimental Allergic Encephalomyelitis in Suckling Lewis Rats: Role of Age and Type of Sensitizing Neuroantigen
Authors: Fujinami Robert S. , Paterson Philip Y.
Published: December 2022

Despite previous reports to the contrary, experimental allergic encephalomyelitis (EAE) can be induced in a variable proportion of suckling Lewis rats after sensitization with neuroantigen-complete Freund's adjuvant (CFA). Three factors have been identified as essential determinants of EAE in suckling animals: a) sensitization via the intracutaneous route, b) age when sensitized, and c) type of neuroantigen injected. For example, animals as young as 4 days of age regularly developed EAE approximately 2 weeks after injection of highly encephalitogenic emulsions of guinea pig spinal cord homogenate-CFA. In contrast, rats as old as 10 days of age injected with soluble syngeneic myelin basic protein (MBP)-CFA developed no evidence of disease whatsoever. Rat spinal cord homogenates and guinea pig MBP gave intermediate results.

Further studies revealed two additional features of EAE induced in suckling rats: a) comparatively less response to the injection of supplemental pertussis vaccine,a known EAE accelerating and potentiating adjuvant in adult animals; and b) a striking paucity of monocytes and histiocytes within the inflammatory reactions at intracutaneous injection sites and the focal nervous tissue cellular infiltrates of animals manifesting typical clinical neurologic signs of the disease. Immaturity of host monocyte-macrophage responses, including processing of soluble antigen, may well account for the contrasting capacities of suckling rats to develop EAE after sensitization to whole cord versus soluble MBP.
Allogeneic Pregnancy as Immunoabsorbent
Authors: Wegmann Thomas G. , Carlson George A.
Published: December 2022

Experiments in the past have indicated that both human and rodent females undergo immune recognition of paternally derived antigens during pregnancy. Although the mechanisms that allow pregnancy to proceed in the face of this process have not yet been clarified, one simple possibility is that the paternal strain antigens are presented in a manner capable of neutralizing the maternal immune response. This predicts that allogeneically pregnant females should be capable of neutralizing passively administered cytotoxic anti-paternal strain antibody, with no consequent harm to the fetus.

In this paper we demonstrate by two different methods that allogeneically pregnant mice rapidly absorb passively administered antibody directed against paternal strain major histocompatibility (H-2) antigens, when compared to control mice pregnant by syngeneic or third party males. The possible sites where this absorption occurs are discussed.
Influence of Inhibitors of Cellular Function on Chemotactic Factor-Induced Neutrophil Aggregation
Authors: O'Flaherty J. T. , Kreutzer D. L. , Showell H. J. , Ward P. A.
Published: December 2022

Chemotactic factors which induce polymorphonuclear neutrophils (PMN) to migrate directionally and release granular enzyme constituents also induce these cells to aggregate. The potency of these factors in inducing aggregation closely parallels their chemotactic and enzyme-releasing potencies. Several reagents known to influence the migratory and degranulatory response of PMN to chemotactins have been examined for their influence on chemotactic factor-induced aggregation of PMN. We have found that ambient temperatures below 37°C, deoxyglucose, and iodoacetic acid inhibit PMN aggregation, whereas sodium cyanide and dinitrophenol have no effect. Inhibitors of microtubules (colchicine and vinca alkaloids) and of protein synthesis (cyclohexamide) had no effect. Cytochalasin B markedly enhanced aggregation.

We conclude that chemotactin-induced aggregation is similar to the other chemotactin-induced PMN functions in the requirements for proper temperature and intact glycolytic pathways; in contrast, however, an intact cytoskeletal microtubular system appears unessential for this response. This may be explained by assuming that the chemotactic factor-induced aggregation of PMN is predominantly a surface membrane-dependent phenomenon.
Viral Protein Synthesis by Tissues from Avian Leukosis Virus-Infected Chickens
Authors: Welt S. , Purchase H. G. , Thorbecke G. J.
Published: December 2022

Chickens of susceptible line 15I were infected at hatching with avian leukosis virus. Viral protein synthesis was detected by showing 14C amino acid incorporation into these proteins by tissues in vitro as shown by autoradiography of viral group-specific (gs) protein immunoelectrophoretic patterns. Synthesis of viral gs proteins p27, p15, and p12 was first detected in all tissues examined of 18-day-old chickens (thymus, spleen, bursa, liver, kidney, testis, ovary, and lung), continued up to 7 to 9 weeks, but could not be detected at 10 weeks, except in lung and genital organs. Grossly leukotic chickens showed gs protein synthesis primarily in the bursa and spleen but, depending on the degree of leukotic infiltration, also in the liver and kidney. Tissues from uninfected controls never showed any gs protein synthesis. The results indicated that all tissues tested were active in viral replication during the initial viremic stage of the disease, whereas during the leukotic stage, lymphoma cells were the predominant cell type active in viral protein synthesis.

Enhanced IgG synthesis in spleen tissue from infected chickens during the viremic stage suggested the presence of a humoral immune response to the virus. Leukotic tissue also appeared to produce both IgM and IgG.
Idiotypes on Galactan Binding Myeloma Proteins and Anti-Galactan Antibodies in Mice
Authors: Mushinski Elizabeth B. , Potter Michael
Published: December 2022

Antibodies with specificity for β1,6 linked D-galactoses were induced in mice by immunization with gum ghatti. Idiotypic antisera were prepared in rabbits and mice by immunization with 8 BALB/c IgA(k), β1,6D-galactan binding myeloma proteins (β6GALBMP). Two kinds of idiotypic sera were obtained: cross-specific sera that reacted with two or more β6GALBMP but not other BALB/c myeloma proteins, and individual idiotypic sera that reacted with only the β6GALBMP used in the immunization. Antibodies with specificity for β1,6 linked D-galactans shared cross-specific idiotypes with β6GALBMP. Only one of seven individual idiotypes associated with β6GALBMP was found on galactan antibodies. Since all β6GALBMP thus far have the same Vk and VH isotype composition the results indicate an extensive heterogeneity among galactan-binding immunoglobulins in BALB/c mice. It is speculated that some of this diversity may arise from somatic rather than germ line gene mutations.
B Cell Tolerance
Authors: Cambier J. C. , Uhr J. W. , Kettman J. R. , Vitetta E. S.
Published: December 2022

The cellular mechanisms and tolerogen dose requirements of hapten-specific unresponsiveness induced in vitro by using 2,4,6-trinitrophenyl human γ-globulin (TNP17HgG) were analyzed in adult and neonatal murine splenocytes. Tolerance induction in both cell populations was found to be independent of non-B cell effects including BAθ-positive cells, Ly 2.2-positive cells, adding or reducing the number of macrophages, and large excesses of HgG. The tolerance induced was specific and not “infectious”, further excluding a role for suppressor T cells. Neonatal splenic B cells were rendered tolerant by doses of TNP17HgG 1000-fold less than those required to produce similar tolerance in splenic B cells from adults. These findings support the concept of functional clonal abortion as a mechanism for producing tolerance to self antigens.
Mechanism for Induction of Anti-DNA Antibodies by Bacterial Lipopolysaccharides in Mice
Authors: Izui Shozo , Zaldivar Nieves M. , Scher Irwin , Lambert Paul-Henri
Published: December 2022

It has been demonstrated previously that there is first a release of DNA in circulating blood and then an induction of anti-DNA antibodies after injection of bacterial lipopolysaccharides (LPS) in mice. These two effects of LPS have been examined in the X-linked B lymphocyte immune-deficient CBA/N mice and LPS-low responder C3H/HeJ mice.

The CBA/N mice and their (CBA/N × DBA/N) F1 hybrid mice injected with 100 µg of Escherichia coli 0111:B4 LPS released DNA in circulating blood in a similar way to the normal CBA/J mice, whereas only the immunologically normal F1 female mice developed anti-DNA antibodies comparable to the CBA/J mice. On the other hand, the LPS-low responder C3H/HeJ mice could neither release DNA in circulating blood nor induce an anti-DNA response to injection of 50 µg Salmonella typhimurium LPS, although they responded to injection of extremely high doses of LPS. However, after reconstitution with spleen cells or anti-θ-treated spleen cells from the LPS-high responder C3HeB/FeJ mice, the C3H/HeJ mice made a substantial anti-DNA response without a significant release of DNA in circulating blood after injection of 50 µg LPS. This indicates that the induction of anti-DNA antibodies by LPS does not require the release of large amounts of DNA in circulating blood.
Inhibition of Antibody-Dependent Cellular Cytotoxicity by Autologous Lymph Node Cells
Authors: Pollack Sylvia B. , Herrick Michael V.
Published: December 2022

A population of lymph node cells that lack the usual T, B, or K cell markers was found to inhibit autologous spleen cells from mediating antibody-dependent cellular cytotoxicity (ADCC) to antibody-coated chicken erythrocytes. Inhibitor cells were not susceptible to treatment with anti-Thy 1.2 or anti-Ig and C; they did not adhere to Sephadex G-10, to nylon wool, or to monolayers of sheep erythrocytes (E) or erythrocytes plus 7S antibody (EA). After a brief (4-min) exposure to 45°C, the ability to inhibit was lost whereas other cellular responses remained intact. ADCC mediated by nonadherent splenic effector cells (presumptive K cells) was highly susceptible to inhibition. Possible mechanisms for and implications of lymphocyte-mediated inhibition of ADCC are discussed.
A Hitherto Undescribed Anomaly in Blood Groups
Authors: Ottenberg Reuben , Johnson Alice
Published: December 2022

Summary and Conclusions

In a case of splenomegaly, hemolysis with a subsequent acute failure of liver function followed a transfusion. The donor's blood contained an abnormal agglutinin active not only against the cells of the patient (who like him belonged to Group III) but also against cells of Groups I, II, and III. It behaved practically like the blood of a foreign species of animal.

It can be inferred that: Donors whose serum agglutinates the cells of the patient (for example “universal” donors, Group I, for patients of Group II, III, or IV) are not necessarily safe.Hemolysis if it occurs is particularly dangerous in diseases of the liver and spleen.It is not enough in transfusion to depend on identity of groups. “Cross tests” of donor's and patient's serum on each other's cells must invariably be done. By this means the danger of hemolysis due to “sub-groups” or rare anomalies can be completely avoided.
On group Specific Substances in Human Spermatozoa
Authors: Landsteiner K. , Levine Philip
Published: December 2022

Experiments have been performed about one year ago with the purpose of examining the presence of group specific substances in human cells other than erythrocytes. We know of a few contributions dealing with this and related subjects. One is a paper on the organs of dogs by von Dungern and Hirschfeld (1), another is by Halpern (2) and a third is a communication by Ashby1 of which only the title is available.

Of particular interest was the behavior of the sperm cells. If they were found to contain group specific substances these should be present in the cells along with those which determine the heredity of the blood groups. The interesting question then, would arise whether the specifically absorbing substances, namely the isoagglutinogens are themselves involved in the inheritance phenomenon, or not. A similar question presents itself with regard to the species specific substances.

The experiments yielded clear-cut results as to the presence of group specific factors in the sperm cells.
The Chemovaccinotherapeusis of Experimental Meningococcus Infection
Authors: Miller A. A.
Published: December 2022

The investigations of Professor Belonovsky have shown that chemical substances in combination with a specific vaccine, injected into the diseased organism, have been transported to the focus of disease by the vaccine, reaching it under the influence of a positive chemotaxis.

In the present study the possibility of a chemovaccinotherapeusis of experimental meningococcus infections has been investigated.

Florand and Fiessinger have already applied the combination of vaccine and chemical substances to the treatment of meningitis, but their iodine-vaccine had no chance of success, as iodine forms a very firm combination with protein.

In my investigations optochin was employed as the chemotherapeutic substance. Optochin is capable in great dilution of inhibiting the growth of meningococcus in vitro and it has been used successfully in the treatment of meningitis by means of lumbar injection (Friedement and Bardachzi).

Some experiments were undertaken to determine the bactericidal power of optochin for the culture, which had been chosen for the tests.
Alterations in Intramembrane Particle Distribution during Interaction of Erythrocyte-Bound Ligands with Immunoprotein Receptors
Authors: Douglas Steven D.
Published: December 2022

Freeze fracture studies have been performed on rabbit pulmonary alveolar macrophages and a nonphagocytic murine lymphoblastoid cell line, PU-5 Fc+, incubated with sheep erythrocytes, sheep erythrocyte-IgG Forssman antibody complex, sheep erythrocyte-IgG Forssman antibody-C complexes and aggregated IgG. Alveolar macrophages show redistribution of intramembrane particles after interaction with E(IgG) and E(IgM)C. The murine lymphoblastoid cell line shows intramembrane particle redistribution consequential to binding of E(IgG) and aggregated IgG. The results demonstrate that after specific immunoprotein receptor-ligand interaction, there is extensive plasma membrane reorganization which results in a redistribution and loss of intramembrane particles. Changes are observed in the protoplasmic face of the plasma membrane after the binding of ligand to the outer membrane surface. The findings suggest that interaction of erythrocyte-bound ligands with specific lymphoid and macrophage plasma membrane receptors leads to a generalized redistribution of integral membrane components in the membrane.
Tolerance to Azobenzenearsonate: Preferential Loss of the Major Normal Cross-Reactive Idiotype
Authors: Benjamin D. C. , Teale J. M.
Published: December 2022

A specific tolerant state was induced in A/J mice to the hapten p-aminobenzenearsonate (Ars) by the injection of deaggregated conjugates of Ars and human γ-globulin (Ars-DHGG). The kinetics of tolerance to Ars-DHGG was found to be identical to that of B cell tolerance to human γ-globulin alone. The ability to produce antibody (anti-Ars) bearing the major normal cross-reactive idiotype was found to be preferentially lost. In addition, recovery of the ability to produce anti-Ars bearing this cross-reactive idiotype was found to be delayed as compared to the total anti-Ars response upon spontaneous loss of tolerance.
Murine T Cells That Lyse Antibody-Sensitized Target Cells
Authors: Lamon E. W. , Williams Barbara R. , Fuson E. W. , Whitten H. D. , Walia A. S.
Published: December 2022

51Cr-labeled sheep erythrocytes sensitized with mouse antibody were lysed by mouse spleen cells and thymocytes. Treatment of the thymocyte donors with cortisone acetate revealed the responsible effector cells to be cortisone-resistant (medullary) thymocytes. Analysis of the thymocyte preparations for cell surface antigens revealed that fewer than 1% of the cells possessed surface Ig detectable by anti-Ig immunofluorescence and 94% of the cells were lysed by anti-thy 1 and C. Less than 0.5% of the cells in the thymocyte preparations were macrophages as determined by their ability to ingest IgG-sensitized erythrocytes. The peak of K cell activity in the thymus occurred at 48 hr after the injection of cortisone. In vitro kinetics experiments revealed that whereas splenic K cell activity reached near maximum levels in 6 to 8 hr (E:T ratio of 10:1), thymocyte-mediated ADCC continued to increase over the entire 72-hr period tested. Lower ratios of spleen cells (2:1 and 1:1) exhibited kinetics similar to thymocytes. Control thymocytes only produced low levels of cytotoxicity by 20 hr, but by 40 and 72 hr in vitro quite significant lysis was produced. Cortisone-resistant thymocytes produced higher levels of cytotoxicity than control thymocytes at all time points. By 72 hr in vitro, the cortisone-resistant thymocytes produced lysis that was equivalent to that produced by spleen cells. The kinetics observed appeared to be the true rate of lysis rather than generation of K cells from precursors in the original population; preincubation of the thymocytes in vitro for 20 hr before the addition of the target cells resulted in no greater lysis than when fresh thymocytes were used. Furthermore, no proliferative response was detectable when thymocytes were incubated with antibody-sensitized target cells.
Lysis of C1q-Coated Chicken Erythrocytes by Human Lymphoblastoid Cell Lines
Authors: Ghebrehiwet Berhane , Müller-Eberhard Hans J.
Published: December 2022

Human lymphoblastoid cells lysed chicken erythrocytes (E) that carried cell surface bound human C1q. Antibody to E (A) was not required for the C1q-dependent reaction. The effect of C1q was inhibited by Fab′2 anti-C1q and by the serum C1q inhibitor. The action of the lymphoblastoid cells was inhibited by anti-metabolites and by pretreatment of the cells with trypsin which is known to destroy their C1q receptor. Lymphoblastoid cell lysate was inactive. The time course of the C1q-dependent lysis was comparable to that of the antibody-dependent cellular cytotoxic reaction of human K-cells. Lysis of EA by human peripheral lymphocytes was enhanced up to 50% by human C1q.
Protein A from Staphylococcus Aureus — A Mitogen for Human T Lymphocytes and B Lymphocytes but Not L Lymphocytes
Authors: Sakane Tsuyoshi , Green Ira
Published: December 2022

Protein A from Staphylococcus aureus (SpA) is known to bind specifically the Fc portion of IgG from several species. In the present paper, SpA was demonstrated to be a potent mitogen for T and non-T lymphocyte populations from both human adult periperal blood and cord blood. SpA was equivalent in mitogenic activity for T lymphocytes as compared to phytohemagglutinin (PHA), and was a much stronger mitogen than concanavalin A.

Experiments separating non-T lymphocytes into B lymphocytes (Fc-) and L lymphocytes (Fc+) by rosetting with IgG-coated sheep erythrocytes demonstrated that SpA can stimulate the proliferation of B lymphocytes but not L lymphocytes.

The addition of mitomycin-treated T lymphocytes to the culture of non-T lymphocytes greatly enhanced the non-T lymphocyte response to SpA, and vice versa. In contrast, enhanced response with either mitomycin-treated monocyte and T lymphocyte combinations or mitomycin-treated monocyte and non-T lymphocyte combinations was not observed. It is suggested that T and non-T lymphocyte synergism is needed for the maximal response to SpA.

Discontinuous bovine serum albumin density gradient fractionation of T lymphocytes demonstrated a population of low density T lymphocytes which responded to SpA better than to PHA, and a population of high density T lymphocytes which responded to PHA better than to SpA. These results indicate that some of SpA-responsive T lymphocytes in human peripheral blood appear to be distinct from a population of T lymphocytes responding to PHA.
Lymphocyte Cytotoxicity in X-Irradiation-Induced Rat Small Bowel Adenocarcinoma
Authors: Stevens R. H. , Brooks G. P. , Osborne J. W. , Hoffman K. L. , Lawson A. J.
Published: December 2022

Hypertonic salt extracts (3 M KCl) of x-irradiation-induced Holtzman rat small bowel adenocarcinomas blocked the in vitro destruction of allogeneic cultured cells of this malignancy by sensitized lymphoid cells obtained from tumor-bearing animals. The protective effects were mediated by a blocking action at both the effector and the target cell level. The extracts were separated into 50% ammonium sulfate soluble and insoluble fractions with the soluble fraction being more effective in blocking the cytotoxic responses through interaction with the lymphoid cells whereas the insoluble one had a greater effect upon tumor target cells. Associated with both fractions was the oncofetal glycoprotein previously identified with the cellular membrane of this x-ray-induced malignancy. Immunoglobulins were identified with the insoluble fraction; some were able to bind the oncofetal protein, thus classifying it as a fetal antigen. The protective effects of the soluble fraction and this neoantigen were found to be citric acid labile, whereas the effects due to the insoluble fraction were unchanged.
Partial Molecular Characterization of the Ly-1 Alloantigen on Mouse Thymocytes
Authors: Durda Paul J. , Shapiro Craig , Gottlieb Paul D.
Published: December 2022

Specific anti-Ly-1.1 serum was used in conjunction with fixed Staphylococcus aureus (Cowan I strain) to precipitate labeled material from NP-40 extracts of mouse thymocytes labeled with 125I by using lactoperoxidase and with NaB3H4 by using galactose oxidase. Thymocytes from the congenic strains C57BL/6-Ly-1a (Ly-1.1-positive) and C57BL/6J (Ly-1.2-positive) were used as sources of labeled antigens and as immune absorbents so that the Ly-1.1 alloantigen could be studied in the absence of genetic background differences. Results of SDS-PAGE under reducing conditions suggest that the Ly-1.1 antigenic determinant(s) may be associated with a glycoprotein of apparent m.w. 67,000 which is labeled by both labeling methods and/or with a component with an apparent m.w. of 87,000 that was labeled with NaB3H4 and galactose oxidase but not with 125I. It is not known whether one or both of these molecular species or an unlabeled species associated with them is the target of the Ly-1.1-specific cytotoxic antibodies. In addition, results of SDS-PAGE under nonreducing conditions suggest that the component with apparent m.w. 67,000 is not involved in intermolecular disulfide bonds.
Correlation between the Ability of Tumor Cells to Resist Humoral Immune Attack and Their Ability to Synthesize Lipid
Authors: Schlager Seymour I. , Ohanian Sarkis H. , Borsos Tibor
Published: December 2022

Agents that increase (certain metabolic inhibitors, chemotherapeutic agents, and x-irradiation), decrease (hormones), or have no effect (hyperthermia) on the susceptibility of line-1 and line-10 guinea pig hepatoma cells to humoral immune attack were studied for their effects on the ability of these tumor cells to synthesize macromolecules. A correlation was found between the drug-induced increase in sensitivity of these cells to antibody-C mediated killing and the loss of their ability to incorporate fatty acids into complex cellular lipids. Similarly, the hormone-induced increase in resistance of the cells to killing was accompanied by an enhancement in complex lipid synthesis by the cells. A correlation between sensitivity to humoral immune killing and complex lipid synthesis by these cells was also observed after the cells were exposed to physical means of insult (x-irradiation or hyperthermia).

No correlation was found between the sensitivity of the cells to antibody-C mediated killing and their ability to synthesize DNA, RNA, protein, or complex carbohydrate, or their capacity for de novo lipid synthesis as measured by incorporation of acetate and glycerol into cellular macromolecules.

The assembly of free fatty acids into complex lipid moieties is therefore proposed to be of fundamental importance for the ability of the tumor cells to resist humoral immune killing.
Identification of Lipids Associated with the Ability of Tumor Cells to Resist Humoral Immune Attack
Authors: Schlager Seymour I. , Ohanian Sarkis H. , Borsos Tibor
Published: December 2022

Certain metabolic inhibitors or chemotherapeutic agents that increase the susceptibility of line-1 or line-10 guinea pig hepatoma cells to humoral immune attack were studied for their effects on the ability of the cells to synthesize lipids. A direct correlation was found between the drug-induced increase in sensitivity to antibody-C mediated killing and the inhibition of the ability of the cells to incorporate acetate, glycerol, and fatty acids into complex cellular lipids. Drug-treated cells recultured in drug-free medium regained their resistance to antibody-C mediated killing; these cells recovered their ability for complex lipid synthesis at this time. Thin layer chromatography of CHCl3:CH3OH lipid extracts from these cells indicated that the drug-induced increase in susceptibility to humoral immune attack correlated with the inhibition of acetate, glycerol, and fatty acid incorporation into cardiolipin and triglyceride in line-10 cells and the inhibition of incorporation of these compounds into cardiolipin alone in line-1 cells. No direct correlation was found between the sensitivity of the cells to humoral immune attack and the ability of the cells to incorporate precursors of lipid synthesis into other lipid moieties (sphyngomyelin, phosphatidyl serine, phosphatidyl choline, phosphatidyl glycerol, or cholesterol esters). The synthesis of cardiolipin and triglycerides, therefore, appears to be associated with the mechanism whereby these tumor cells resist antibody-C mediated killing.
Evidence for Antibody Activity against the Receptor for IgE in a Rabbit Antiserum Prepared against IgE-Receptor Complexes
Authors: Conrad Daniel H. , Froese Arnold , Ishizaka Teruko , Ishizaka Kimishige
Published: December 2022

A rabbit antiserum prepared against rat IgE-receptor complexes was examined for evidence of antibody activity against receptors for IgE. The antibodies were specifically purified by absorption onto and subsequent elution from intact rat basophilic leukemia (RBL) cells and rendered specific for mast cells by absorption with mast cell-depleted rat peritoneal exudate cells. Surface-labeled RBL cells were extracted with Nonidet P-40 (NP-40) and the soluble extract was incubated with the purified antibodies; the components bound by the antibodies were analyzed on 10% SDS polyacrylamide gels. The major component bound by the antibodies had a relative mobility identical to that of the receptor for IgE. Confirmation that this component was indeed the receptor for IgE was shown in that this receptor was progressively removed from the NP-40 extract by increasing amounts of the purified antibodies. The ability of these purified antibodies to interact with the receptor was strongly inhibited when the receptor was complexed with IgE; in addition, the purified antibodies demonstrated little activity against purified IgE-receptor complexes. These results suggested that the antigenic site(s) interacting with the purified antibodies were either in or closely adjacent to the IgE-binding site on the receptor. In contrast, the original antiserum, after removal of anti-IgE antibodies by absorption with IgE covalently linked to Sepharose, demonstrated significant activity against the purified IgE-receptor complexes. This suggested that antibodies against sites on the receptor distant from the IgE-combining site were present in the original antiserum. These sites were probably buried in the membrane and thus lost in the antibody purification procedure.
The Distribution and Relative Immunogenicity of Calf α-Crystallin Antigenic Determinants on Different Subunits
Authors: Manski W. , Malinowski K.
Published: December 2022

In the native α-crystallin molecule, 45.9% of all reactive antigenic determinants were found to be located on SH-containing subunits. Of these, the majority (35.3%) were reaggregation dependent, and 10.6% were reactive on monomeric subunits. By contrast, only 10.9% of all antigenic determinants were located on SH-free subunits, and the ratio of aggregation-dependent determinants (4.4%) to those of monomeric subunits (6.5%) was reversed compared to SH-containing subunits. Among all antigenic determinants reactive in native α-crystallin, 44.1% were dependent on the presence of both types of subunits. These data indicate that the antigenic determinants requiring subunit interaction were formed from SH-containing and SH-free subunits in a ratio of 1:1. Direct analysis showed that in the α-crystallin molecule, the ratio of these subunits is 2:1. The experiments indicate that some conformations of subunits in the native molecule persist in separated subunits. The relative immunogenicity of each type of antigenic determinant expressed as the ratio of the percentage of the determinant reactive in the native calf lens α-crystallin to the percentage of corresponding antibodies induced by native α-crystallin was found to be close to 1.
Immunoregulatory Function of Specific IgG
Authors: Michael J. Gabriel , Bernstein I. Leonard
Published: December 2022

Nonatopic subjects immunized with short ragweed developed both reaginic IgE and blocking IgG antibodies to the antigen. Administration of 250 µg-specific ragweed IgG antibodies 1 week before, simultaneously, or up to 2 weeks after start of immunization with ragweed extract abrogated the production of anti-ragweed reaginic antibodies which was measured by skin reactivity. Formation of specific ragweed IgG antibodies was unaffected by IgG treatment. These results indicate selective regulation of an immune response in humans by IgG.
Ontogeny of IgE-Bearing Lymphocytes in the Rat
Authors: Ishizaka Kimishige , Ishizaka Teruko , Okudaira Hirokazu , Bazin Hervé
Published: December 2022

IgE-bearing lymphocytes were detected by immunofluorescence in the spleen of neonatal Hooded Lister strain rats within 24 hr after birth. The same cells were detected in the bone marrow as early as the 4th day after birth. Both fetal spleen and liver obtained 1 day before birth contained IgM-bearing cells but no detectable IgE-bearing cells. The proportion of IgE-bearing cells in the spleen and bone marrow increased during the neonatal period and reached an adult level within 3 to 4 weeks after birth. In adult Hooded Lister rats, IgE-bearing cells were 3 to 6% of total spleen cells and 1.5 to 2.2% of bone marrow cells. Most of the IgE-bearing cells from both newborn and adult animals carried IgM determinants on their surface. Capping experiments showed that µ chain determinants and µ chain determinants belonged to separate molecules. IgG2a-bearing lymphocytes were detected in the neonatal spleen as early as the 4th day after birth, but a significant number of these cells was not detected in the bone marrow until the 4th week. In newborn spleen the percentage of IgE-IgM double bearing cells was higher than that of IgG2a-bearing cells.
Inactivation of Antigen-Responsive Clones with Antisera Specific for IgM or IgD
Authors: Ligler F. S. , Cambier J. C. , Vitetta E. S. , Kettman J. R. , Uhr J. W.
Published: December 2022

The present study examines the isotype of cell surface immunoglobulins involved in triggering a response to thymus-dependent and thymus-independent antigens. Antibodies to immunoglobulin isotypes were used to block the in vitro interaction of receptor IgD and IgM with antigen. The results indicate that both IgD and IgM are necessary to trigger a response to TNP-SRBC but that only IgM is required for responsiveness to TNP-Brucella. Limiting dilution studies indicate that the inhibition of the immune response by antibody occurs at the level of precursor activation and suggest that there is no effect of antibody on subsequent proliferation of the daughter cells.
Latent Expression of a Cγ Gene
Authors: Mandy William J. , Strosberg A. Donny
Published: December 2022

Rabbit 136 represented the first instance whereby more than two alleles for any immunoglobulin locus are detected phenotypically in a single animal. This was reported as the acquisition of the a2 and b6 markers in the animal nominally typed a1a3, b4b5 after intensive immunization. Follow-up studies indicate that another immunoglobulin marker, notably d11, also appeared in the postimmune serum. The d11 specificity was detected serologically by the hemagglutination-inhibition assay and chemically by analysis of the CNBr digestion products involving a strategic methionine.
B Lymphocyte Precursors in Human Bone Marrow: An Analysis of Normal Individuals and Patients with Antibody-Deficiency States
Authors: Pearl Elliott R. , Vogler Larry B. , Okos Anthony J. , Crist William M. , Lawton Alexander R. , Cooper Max D.
Published: December 2022

Lymphoid cells containing cytoplasmic IgM but lacking stable surface IgM are believed to be the direct precursors of B lymphocytes. We have characterized these pre-B cells in the bone marrow of normal individuals and patients with a variety of immunoglobulin deficiencies or hematologic disorders by using immunofluorescence and autoradiography. Pre-B cells comprised 5.8 ± 5.7% of lymphoid cells in normal bone marrow. Eleven patients with infantile X-linked agammaglobulinemia (X-LA) lacked B lymphocytes but had a normal frequency (3.8 ± 3.6%) of bone marrow pre-B cells. A smaller proportion of marrow pre-B cells from patients with X-LA were engaged in spontaneous DNA synthesis than was found for normal controls. In individuals other than the group with X-LA, the number of circulating B cells was positively correlated with the frequency of marrow pre-B cells. These results indicate that patients with X-LA have a defect in maturation of pre-B cells, and suggest that some patients with acquired B lymphocyte deficiency may have lost the capacity to generate pre-B cells from stem cells.