bluuebunny/crossref_metadata_2025 huggingface dataset:
Stimulation of Prostaglandin Synthesis with Resultant Bone Resorption by the Classical and Alternative C Pathways
Authors: Sandberg Ann L. , Raisz Lawrence G. , Goodson J. Max , Mergenhagen Stephan E.
Published: December 2022

In vitro studies suggest that C activation may contribute to the bone loss associated wath certain inflammatory conditions. Rat fetal long bones (radii and ulnae), prelabeled in utero with 45Ca, were cultured in the presence of rabbit serum either containing or lacking C activity, and resorption was detected by the efflux of 45Ca into the media. In the present studies, serum concentrations were chosen which did not themselves support bone destruction since previous investigations demonstrated resorption in the presence of high concentrations of C sufficient serum. However, in these limited serum cultures, resorption could be initiated by the addition of activators of either the classical or alternative C pathways. Rabbit anti-rat erythrocyte IgG, obtained by DEAE-cellulose chromatography, and 19S and 7S fractions, obtained by Sephadex G-200 column chromatography, initiated resorption in the presence of C, whereas, non-tissue-related immune complexes were inactive.
Mechanism of Action of the Chemotactic Factor Inactivator of Human Serum
Authors: Ward P. A. , Ormrod J. C. , Ozols J.
Published: December 2022

The chemotactic factor inactivator (CFI) irreversibly inhibits the leukotactic activity of complement C3 and C5 fragments, and the activity of leukotactic factors produced in bacterial cultures. In order to elucidate the mechanism of inhibition, CFI was purified from human serum and its interaction with synthetic peptides investigated. Kininase activity was present in all CFI preparations, as revealed by bioassay of bradykinin which had been pretreated with CFI. Amino acid analysis of the inactivated bradykinin preparations revealed the presence of free arginine, proline, phenylalaninine, glycine, and serum. Incubation of CFI with the highly active chemotactic peptide Met-Leu-Phe or with its amide derivative, resulted in a concomitant loss of the leukotactic activity and the simultaneous appearance of the corresponding free amino acids. The highly potent chemotactic peptide formyl-Met-Leu-Phe, however, was completely resistant to CFI hydrolysis, as assessed by bioassay (chemotaxis) and amino acid analysis methods.
C3bINA Accelerator: A Serum Cofactor for C3b Inactivator Necessary for the Full Expression of Its Activity
Authors: Whaley Keith , Ruddy Shaun
Published: December 2022

C3b inactivator (C3bINA) may be measured as activity in a hemolytic assay or as protein by radial immunodiffusion. The observation that purified C3bINA has fewer units of activity per microgram or C3bINA protein than C3bINA in unfractionated serum is accounted for by the presence in serum of a cofactor which accelerates C3bINA-dependent C3 inactivation.

Sephadex G-200 chromatography of human serum yields a single peak of C3b-inactivating activity coincident with immunochemically detectable C3bINA. A second area of C3b-inactivating activity appears on the descending limb of the 19S peak if fractions are assayed in a system containing subthreshold quantities of purified C3bINA. This activity, which has little or no C3b-inactivating activity by itself, but enhances the activity of purified C3bINA, has been termed the C3b inactivator accelerator (A·C3bINA). It is separable from C3bINA on Biorex-70 at pH 7.5, is destroyed by trypsin, but is resistant to di-isopropyl phosphofluoridate.
Identification and Purification of Immunosuppressive Activity in the Urine of Rats and a Human Patient Treated with Niridazole
Authors: Lucas Samuel V. , Daniels Jerry C. , Schubert Richard D. , Simpson Janney M. , Mahmoud Adel A. F. , Warren Kenneth S. , David John R. , Webster Leslie T.
Published: December 2022

Administration of the antischistosomal compound niridazole to mice, guinea pigs, and humans results in the suppression of several manifestations of cell-mediated immunity. Sera from animals treated with niridazole blocked the in vitro production of migration inhibitory factor (MIF) while niridazole itself was inactive, suggesting that these effects are caused by water soluble mediators. We now report that crude extracts prepared from the urine of rats and a patient receiving niridazole, but not from pretreatment control urine, similarly suppress antigen-induced inhibition of migration of peritoneal exudate cells from sensitized guinea pigs. With immunosuppressive activity monitored by the direct MIF assay, combined solvent extraction and chromatographic techniques were used to fractionate immunosuppressive activity from the urine of niridazole-treated rats and the patient; the most active fractions, purified about 100- to 1000-fold as compared to methanol-water extracts of dried voided urine, inhibited MIF production at 0.1 to 0.01 ng/ml of assay mixture. These purified fractions also showed immunosuppressive activity by an in vivo assay wherein doses as low as 1 µg/kg injected intravenously (i.v.) into mice suppressed cell-mediated granuloma formation around Schistosoma mansoni eggs. Identically purified fractions prepared from urine of rats and the patient before they received niridazole showed no immunosuppressive activity either in the MIF or in the granuloma assay systems.
Ir-Gene Control of Antibody Class Production
Authors: Kolb Hubert , Keck Klaus , Momayezi Massoud , Schicker Cornelia , Trissl Dorothea
Published: December 2022

The antibody response in mice to DNP-insulin is under Ir-gene control. The Ir gene defects in two strains have been analyzed. In both cases the IgG immune response was impaired whereas the IgM response was not affected. One H-2 gene haplotype was characterized by lack of IgG response, independent of the immunization protocol. A second H-2 haplotype manifested a low response of IgG after immunization with Bordetella pertussis as an adjuvant but a high response after complete Freund's adjuvant. It is proposed that a low level of T cell help induces the production of IgM antibodies, intermediate levels allow few IgG clones to develop, and high levels induce a heterogeneous IgG response.
Serologic Diagnosis of Lymphogranuloma Venereum by Counterimmunoelectrophoresis with a Chlamydia Trachomatis Protein Antigen
Authors: Caldwell Harlan D. , Kuo Cho-Chou
Published: December 2022

Previously a protein antigen was identified which was common to members of the species Chlamydia trachomatis and to which antibody could be demonstrated in human sera by two-dimensional immunoelectrophoresis. The antigen has been purified by immunoadsorption chromatography, and an exploration undertaken to determine whether the detection of antibody to this antigen by counterimmunoelectrophoresis would be useful for the diagnosis of C. trachomatis infections. Antibody was found in 43 of 45 (96%) sera from patients with lymphogranuloma venerum proven by either isolation or other serologic methods. No positive reactions were observed in 50 patients with nongonococcal urethritis, 38 of whom had evidence of antibody as measured by microimmunofluorescence with whole organism antigen. Sera were also negative from 50 normal individuals, 10 persons with gonorrhea, and 2 patients with psittacosis. Counterimmunoelectrophoresis with purified C. trachomatis protein antigen appears to be a useful serologic test for diagnosis of lymphogranuloma venereum.
Activation of the Alternative Pathway by Pneumococcal Cell Walls
Authors: Winkelstein Jerry A. , Tomasz Alexander
Published: December 2022

The present studies were performed in order to identify the pneumococcal subcellular component responsible for activating the alternative pathway.

Purified pneumococcal cell walls were able to activate the alternative pathway at a concentration as low as 5 µg/ml and were more active than crude cell walls, which in turn were more active than the whole organism. Purified pneumococcal cell membranes also were able to activate the alternative pathway but had less than 10% of the activity of the purified walls. Thus, the cell wall appears to play a major role in pneumococcal activation of the alternative pathway.

Pneumococcal cell walls containing ethanolamine were as effective as cell walls containing choline in activating the alternative pathway. Since C-reactive protein binds specifically to the phosphorylcholine residue of pneumococcal C-polysaccharide, it is unlikely that pneumococcal cell walls must combine with C-reactive protein in order to activate the alternative pathway.
Spleen-Derived Mononuclear Cell Chemotactic Factor in Malaria Infections: A Possible Mechanism for Splenic Macrophage Accumulation
Authors: Wyler David J. , Gallin John I.
Published: December 2022

A hallmark of malaria and several other intravascular infections is the development of splenomegaly associated with splenic macrophage hyperplasia. These macrophages do not arise as a consequence of in situ proliferation of preexisting splenic macrophages, but rather from blood monocytes. To investigate the mechanism of accumulation of blood monocytes in the spleen during the development of splenomegaly, we studied mice and monkeys infected with malaria and mice treated with methyl cellulose. Employing a modified Boyden chamber chemotaxis assay, extracts of washed spleen cells of malaria-infected but not control mice or monkeys were found to contain a factor chemotactic for homologous and human peripheral mononuclear cells. In mice, this factor was present within 4 hours of infection, persisted for the duration of infection, and anteceded a detectable increase in splenic macrophage number by 48 hr. Extracts of lysed parasitized erythrocytes contained no chemotactic activity. Supernatants from in vitro cultures of spleen cells from malaria-infected but not control mice also contained mononuclear cell chemotactic activity. The spleen-derived chemotactic factor was present in nonadherent cells, was water soluble, heat labile, trypsin sensitive and had an estimated m.w. of 80,000 daltons (Sephadex G-150). It contained no serine esterase activity. These observations suggest that infection with malaria and administration of other agents inducing splenomegaly result in the in vivo elaboration of an endogenous spleen-derived mononuclear cell chemotactic factor. This factor, presumably derived from lymphocytes, may be responsible for or contribute to the trapping of blood monocytes in the development of macrophage hyperplasia in the spleen.
The Mononuclear Cell in Human Blood Which Mediates Antibody-Dependent Cellular Cytotoxicity to Virusinfected Target Cells
Authors: Melewicz F. Michael , Shore Steven L. , Ades Edwin W. , Phillips Don J.
Published: December 2022

Studies were carried out to determine whether the mononuclear cell in human blood which mediates antibody-dependent cellular cytotoxicity (ADCC) to herpes simplex virus (HSV)-infected target cells has surface Fc receptors which participate in the reaction.

The F(ab′)2 fragment of human IgG antibody was inactive both in ADCC and in complement-mediated cytolysis, but retained the capacity to neutralize infectious virus, to agglutinate erythrocytes coated with viral antigens, and to bind to the surface of virus-infected cells. Treatment of sensitized virus-infected target cells with staphylococcus protein A, which has affinity for the Fc epitope of IgG, strongly reduced their susceptibility to lysis by ADCC in a dose-dependent relationship. These findings indicate that the Fc portion of IgG antibody to the virus is necessary for cytotoxicity.

Treatment of blood mononuclear cells with either heat-aggregated γ-globulin or HSV immune complexes inhibited effector cell activity. The presence of “third party” cellular immune complexes also strongly inhibited ADCC. Adsorption of mononuclear cells to plastic surfaces coated with soluble third party immune complexes resulted in a significant reduction in effector cell activity.

These findings demonstrate that the ADCC effector cell possesses surface Fc receptors which are utilized in the ADCC reaction. The presence of Fc receptors on the surface of the effector cell indicates that it is a K cell rather than a null cell.
Immunologic Characteristics in Relation to High and Low Leukemogenic Activity of Radiation Leukemia Virus Variants
Authors: Haran-Ghera Nechama , Ben-Yaakov Miriam , Peled Alpha
Published: December 2022

Infection of adult C57BL/6 mice with variants of the radiation leukemia virus resulted in variable leukemia incidence. One variant, designated D-RadLV, induced lymphatic leukemia in 0 to 25% of mice after virus inoculation directly into the thymus of young adult mice. The leukemia incidence could be increased to 80 to 100% by host exposure to x-rays. The second variant, A-RadLV, induced lymphatic leukemia in 80 to 100% of similarly inoculated mice without the need for additional radiation treatment.

Adult mice were inoculated with D-RadLV or A-RadLV. Both variants reduced the immune response to sheep erythrocytes whereas only D-RadLV had an immunosuppressive effect after immunization with a thymus-independent immunogen polyvinyl-pyrrolidone (PVP). Results of transfer experiments indicated that the immunosuppressive effects were expressed at the immunocompetent cell level. Thymusderived cells were affected by A-RadLV since their immunocompetent function was impaired, whereas D-RadLV affected the marrow cell population of immunocytes. Exposure of D-RadLV-inoculated mice to x-rays induced functional impairment of both thymus and marrow cells.

Since the radiation leukemia virus induces “T” lymphatic leukemia it could be proposed that the initial tropism of the virus to thymocytes would lead to high leukemia induction potential, whereas virus tropism to bone marrow cells would yield a low leukemia incidence. The coleukemogenic effect of x-rays could perhaps be related with its capacity to alter and introduce a change in virus-lymphoid cells interaction.
Ontogeny of the Antibody-Forming Cell Line in Mice
Authors: Rosenberg Yvonne J. , Parish Christopher R.
Published: December 2022

The ontogeny of Ig, FcR, and CR-bearing cells in liver and spleen has been followed by using rosetting procedures. These studies demonstrated a sequential appearance of surface receptors during development. Two types of Ig+ cells could be distinguished according to their rosette morphology and adherence to carbonyl iron: 1) an adherent cell which bound few erythrocytes was found predominantly in fetal liver from 13 days gestation and 2) a nonadherent cell which bound larger numbers of erythrocytes appeared in small numbers in fetal liver from day-16 gestation but represented the major Ig+ cell type after birth. Changes in the proportions of receptor-bearing populations occurred at two particular periods during ontogeny. The first was at birth, where an increase in the proportion of FcR+ cells occurred and the proportion of type 2 Ig+ cells rose rapidly. This probably represented the first appearance of FcR+ B lymphocytes even though cells bearing FcR were detected in fetal liver of all ages (days 12 to 18). The second period was around 10 days after birth when the proportion of Ig+ cells again increased concomitant with the appearance of CR+ nonadherent cells.
Identification of Human Peripheral T Cell Antigens
Authors: Koshiba Hirofumi , Ishii Yoshifumi , Yamanaka Noboru , Kikuchi Kokichi
Published: December 2022

A new type of differentiation antigens on human T cells was demonstrated by using a heterologous anti-human T cell serum (ATS). This type of antigen, referred to as human peripheral T cell antigen (HPTA), was found on peripheral T cells and medullary thymocytes, but not on cortical thymocytes and B cells. The percentage of ATS-reactive lymphocytes in human peripheral lymphoid organs was correlated with that of cells rosetting with sheep erythrocytes, but contrasted with the number of B cells defined by the presence of a complement (C) receptor or by rabbit anti-human B cell serum (ABS). ATS also reacted with T cells purified by nylon fiber column filtration but ABS did not. Chronic lymphocytic leukemia cells rosetted with either sheep erythrocytes or erythrocyte-antibody-complement complexes were lysed by ATS and ABS, respectively. Mitogenic responses of blood lymphocytes to phytohemagglutinin (PHA) and concanavalin-A (Con A) were abrogated by treating them with ATS and C, whereas ABS suppressed only their response to Con A. Although numerous thymus cells rosetted with SRBC, only 14% were reactive with ATS. Quantitative absorption studies demonstrated that HPTA content of the thymus cells was much lower than that of lymph node cells.

Anatomical localization of ATS-reactive lymphocytes in human lymphoid organs studied by immunofluorescence indicated that they were present in the thymus-dependent paracortical areas of lymph node and in the medullary region of thymus. ABS, on the other hand, did not stain thymocytes but reacted selectively with the cells located in the lymphoid follicles of lymph node. These data, together with that from cell suspension studies, confirmed that HPTA were shared between medullary thymocytes and peripheral T cells.
Immune Competence of Splenic Lymphocytes Following Graft-vs-Host Disease in Mouse Allogeneic Radiation Chimeras
Authors: Urso Paul , Gengozian Nazareth
Published: December 2022

The abnormal immune response of long-term mouse allogeneic chimeras is reflected by qualitative deficiencies in either T or B lymphocytes. The present study was undertaken to determine if a relationship existed between the severity of graft-vs-host disease (GVHD) that these animals had experienced and a functional defect in either the T or B cell population. The in vitro PFC response of chimera spleen cells to sheep red blood cells (SRBC) was evaluated in the presence of normal T or B lymphocytes 4 to 8 months after marrow transplantation and well beyond the GVHD period. In an analysis of several different allogeneic radiation chimeras, our results showed no relationship between the severity of GVHD experienced and the immunologic capacity of either T or B cells. Thus, different chimera combinations showing similar degrees of GVHD were functionally deficient in one or the other of these two cells types or both with no apparent predilection for abnormality in either population. In examining the quantitative in vitro PFC response to sheep RBC by spleen cells from individual chimeras, we found that the number of PFC formed was related to the severity of GVHD experienced by that animal. A general relationship between severity of GVHD and PFC capacity may also exist between chimeras of different genetic combinations. However, this relationship is not precise since gross exceptions occur. Our results, although documenting further the qualitative abnormalities in T and/or B lymphocytes of radiation chimeras, do not reveal the factor or mechanisms by which these cells are made unresponsive. It is suggested that the tolerance-inducing mechanism of these animals, whether it be humoral blocking factors or suppressor cells, is in some way interfering with the collaboration of T and B cells for antibody production.
Genetic Factors Influencing Mouse Type-C RNA Virus Induction by Naturally Occurring B Cell Mitogens
Authors: Phillips S. Michael , Stephenson John R. , Aaronson Stuart A.
Published: December 2022

The present studies demonstrate that xenotropic type C virus is efficiently released in response to lipopolysaccharide by spleen cells of a wide variety of inbred mouse strains. Lipopolysaccharide-mediated virus release primarily involves B lymphocytes and is in part genetically determined. Virus release can also be efficiently stimulated by other naturally occurring B cell mitogens, including Nocardia water soluble mitogen, and PPD. The evidence indicates that these agents act synergistically with halogenated pyrimidines, but not with each other, to cause virus release. These results indicate that B cell mitogens act to release virus by a mechanism that differs from that of halogenated pyrimidines.
Antigenic Specificity of Opsonophagocytic Antibodies in Rabbit Anti-Sera to Group B Streptococci
Authors: Baltimore Robert S. , Kasper Dennis L. , Baker Carol J. , Goroff Diana K.
Published: December 2022

An opsonophagocytic assay has been developed which requires human polymorphonuclear leukocytes, immune serum, and complement for optimal killing of Group B streptococci. Only with all three of these components was killing of greater than 1.0 log10 of the initial inoculum achieved, using rabbit antisera directed to homologous strains of each of the five known serotypes of Group B streptococci. Titers of specific antisera which opsonized the strains and resulted in τ1 log10 reduction of colony-forming units, ranged from 1:100 (serotype Ib) to 1:3200 (serotype Ia). Cross-reactions between serotype-specific sera and heterologous strains were seen in certain instances. Type Ic strain and serotype Ic antiserum demonstrated cross-reactions with types Ia and Ib which were explainable by known shared antigens among these types. The only other cross-reaction which resulted in τ1 log10 reduction in colony-forming units was when unabsorbed antiserum to strain Ia was used to opsonize a strain of serotype III. Opsonization of 10 serotype III strains was demonstrated with a single type III antiserum. Killing of nine of these strains required polymorphonuclear leukocytes, complement, and antiserum, but one strain, D136C, the reference strain, could be killed (τ1 log10 reduction in colony-forming units) without either complement or specific antiserum.

Inhibition studies were performed utilizing large m.w. polysaccharide antigens extracted from each serotype. These antigens inhibited opsonization of homologous strains by homologous antisera with 50% inhibition points ranging between 0.5 and 4 µg.
Suppression of the Humoral Immune Response by Plasmacytomas: Mediation by Adherent Mononuclear Cells
Authors: Kolb Jean-Pierre , Arrian Shelley , Zolla-Pazner Susan
Published: December 2022

Mice bearing plasmacytomas have a severely impaired ability to mount a primary immune response; T cells from these mice, however, appear by both in vivo and in vitro criteria to function normally. This unusual pattern of immunodeficiency appears to be mediated by a regulatory cell found in the spleens and peritoneal cavities but not in the lymph nodes or thymuses of mice bearing plasmacytomas. The number of cells with suppressor activity in the spleens of plasmacytoma-bearing mice is directly proportional to the size of the subcutaneous tumor borne by the host. These cells are capable of suppressing antibody production in in vitro cultures of normal spleen cells but have no demonstrable effect on the ability of normal spleen cells to proliferate in vitro in response to phytohemagglutinin or 8-Br-guanosine-3′, 5′-monophosphate (T and B cell mitogens, respectively). Characterization of the suppressor cell population on the basis of its cell surface properties, its radioresistance, its morphology, and its ability to adhere to various solid matrices suggest that these cells are adherent mononuclear cells. These data support the concept that plasma cell tumors indirectly induce an impairment in the humoral immune response of their hosts by stimulating the expression of regulatory functions in a population of splenic and peritoneal macrophages.
Differential Effects of Interferon on the Expression of H-2K, H-2D, and Ia Antigens on Mouse Lymphocytes
Authors: Vignaux Françoise , Gresser Ion
Published: December 2022

Lindahl and co-workers (1, 2) have reported that mouse interferon enhances the expression of H-2 antigens on the surface of murine leukemia L 1210 cells (H-2d) and normal mouse thymocytes and splenic lymphocytes from DBA/2 (H-2d) or C57 BL/6 (H-2b) mice (3) and presented evidence that interferon was the responsible factor (1, 3). Furthermore, inoculation of DBA/2 mice with potent partially purified mouse interferon preparations (or viral or nonviral inducers of endogenous interferon) was also accompanied by a marked increase in the expression of H-2 antigens on the surface of thymocytes and splenic lymphocytes (4). The availability of restricted antisera against determinants controlled by subregions of the H-2 complex (Table I) has permitted us to show that interferon enhances the expression of H-2K and H-2D antigens, while affecting only slightly, or not at all the expression of Ia antigens on mouse lymphocytes.
Interaction between Taenia Taeniaeformis and the Complement System
Authors: Hammerberg Bruce , Williams Jeffrey F.
Published: December 2022

Factors present in the cystic bladder fluid of metacestodes of Taenia taeniaeformis and released by these parasites maintained in vitro were shown to interact with the C system in vitro and in vivo. This interaction resulted in the depletion of hemolytic C in normal sera of several species, the generation of anaphylatoxin-like activity in vitro, the conversion of C3, and the production of vascular permeability changes in vivo. The substances appeared to initiate C fixation nonimmunologically via both the alternative and classical pathways. Intravenous administration of parasite factors in rats led to profound depression of circulating C levels and the rats did not become refractory to this effect after repeated dosing over a 3-week period. Circulating levels of hemolytic C were not altered in infected rats over the first 8 weeks of infection. C-dependent inflammatory responses in the skin of rats were inhibited in animals given i.v. doses of parasite factors. The possibility is raised that local consumption of C around the metacestode in vivo could contribute to its successful evasion of inflammation and immune rejection during infection.
The Maintenance and Regulation of Serum Antibody Levels: Evidence Indicating a Role for Antigen Retained in Lymphoid Follicles
Authors: Tew John G. , Mandel Thomas
Published: December 2022

The synthesis of anti-human serum albumin (HSA) antibody was induced in the absence of added antigen in lymphoid cells obtained from CBA mice that had been immunized weeks to months earlier. This response was restricted to cells or tissue fragments obtained from lymphoid organs in the lymphatic drainage path of the original site of antigen administration. Injection of 125I-HSA revealed that retained antigen was likewise restricted to the spleen and draining lymph nodes and that it was present in these organs for long periods of time. Autoradiography indicated that within the limits of detection, antigen was further restricted to cells within the lymphoid follicles and was not retained in macrophages of the medulla, the subcapsular sinus, or the superficial cortex. Small fragments of lymph nodes placed in organ culture showed that the spontaneous response corresponded with the presence of follicular antigen. The more antigen present in a fragment the greater the spontaneous response. The belief that retained radioactivity represented active immunogen, even 7 weeks after injection, was supported by the observation that it could be specifically depleted from the lymphoid follicles by injecting nonradioactive HSA. On the basis of these data it appears likely that antigen retained in lymphoid follicles remains active for prolonged periods and stimulates antibody synthesis whenever antibody levels decline. Thus, retained follicular antigen could play a major role in the mechanism which maintains and regulates serum antibody levels for months or years after immunization.
Immune Response in Guinea Pigs to Two Different Lipid Conjugates of Bovine Serum Albumin
Authors: Singh Suraj B. , Leskowitz Sidney
Published: December 2022

A lipid conjugate of BSA was made by coupling dodecylamine to the COOH groups of BSA (DA-BSA). Studies on the immunogenicity of this material and one made by coupling dodecanoic anhydride to NH2 groups (D-BSA) demonstrated that cell-mediated immunity could preferentially be generated by lipid conjugation to an antigen as measured by delayed skin reaction, in vitro blast transformation, and antibody formation. DA-BSA was found superior by both in vivo and in vitro tests. Both the conjugates retained the ability to suppress delayed type hypersensitivity, but only DA-BSA retained the ability to elicit optimum skin reaction in a sensitized animals and to precipitate anti-BSA antibody.
The Regulation of IgM Production in the Chick: Roles of the Bursa of Fabricius, Environmental Antigens, and Plasma IgG
Authors: Rao D. S. V. Subba , McDuffie F. C. , Glick Bruce
Published: December 2022

Previous studies by us have shown that preventing development of the bursa of Fabricius by dipping fertile eggs in testosterone propionate (TP) on the third day of incubation results in elevated levels of IgM in plasma beginning 2 to 4 weeks after hatching. In the present study we have found that such chicks repeatedly immunized with sheep red blood cells (SRBC) produce increasing amounts of IgM antibody but no IgG antibody, in contrast to normal birds which show increasing antibody responses only in the IgG class. TP birds raised in a pathogen-free (PF) environment from hatching until 10 weeks of age have significantly lower levels of IgM than TP birds raised in a conventional environment. The IgM levels approach those of the latter birds at 15 weeks, 5 weeks after being returned to a conventional environment. Further experiments with infusion of normal and immune IgM and IgG into normal birds demonstrated some enhancement of IgM levels by IgM but marked depression of levels of immunoglobulin and antibody in both classes by IgG. The results strongly support the hypothesis that elevated levels of IgM in bursaless, TP-treated birds result from immune responses to environmental antigens unregulated by feedback from IgG. They provide further evidence that the bursa is essential for the normal switch from IgM to IgG production but not for the development of lymphoid cells producing IgM.
Transfer of Concanavalin A Between Responding Lymphocytes and Syngeneic Stimulating Cells in Cell-Mediated Mitogenic Response
Authors: Ozato Keiko , Ebert James D.
Published: December 2022

We have studied the mechanism of the cell-mediated mitogenic response (CMMR), in which proliferative responses are generated in mouse T lymphocytes co-cultivated with syngeneic mitomycin C-treated spleen cells (Mito-SP), when either responder or stimulator cells are briefly pretreated with concanavalin A (Con A) under nonmitogenic conditions. We present evidence that an intact membrane of the stimulator cells is required in CMMR, since the response was abolished by fixation, or by freezing and thawing of stimulator cells.

The fate of cell-bound Con A was studied by tracing 125I-Con A bound to either stimulator or responder. A lymphoblast that developed from untreated CRT stimulated by 125I-Con A-coated syngeneic Mito-Sp carried about 105 molecules of 125I-Con A. The amount of Con A released into the culture medium was not sufficient for inducing a mitogenic response by itself, nor to bind to cells at the level found in CMMR, suggesting that 125I-Con A was transferred directly from labeled cells to unlabeled cells. Transferred 125I-Con A found in lymphoblasts was undegraded intact Con A, as demonstrated by gel electrophoresis. Autoradiography allowed visualization of the movement of 125I-Con A from stimulator to responder cells within 60 min of cell contact.
Natural Killing in Immunodeficient Patients
Authors: Koren Hillel S. , Amos D. Bernard , Buckley Rebecca H.
Published: December 2022

Natural killing (NK) capacity was evaluated in peripheral blood mononuclear cells from 14 patients with well defined primary immunodeficiency disorders and compared with the activity of those cells in antibody-dependent cell-mediated cytotoxicity (ADCC) assays against antibody-coated erythrocyte (killed primarily by monocytes) and lymphoid or tumor targets (killed exclusively by lymphoid cells). A selective inability to lyse antibody-coated lymphocyte targets was observed with cells from patients with x-linked agammaglobulinemia, suggesting the involvement of either a different lymphocyte subpopulation or membrane receptor for NK and ADCC, or that a different functional susceptibility exists for the two types of killing. The only immunodeficiency state in which lymphocyte NK activity was found to be lacking was severe combined immunodeficiency disease.
Expression of Murine Ia Antigens during Embryonic Development
Authors: Delovitch Terry L. , Press Joan L. , McDevitt Hugh O.
Published: December 2022

An immunochemical analysis of the kinetics of appearance of Ia antigens during embryonic development was performed. Ia antigens first appear on the surface of embryonic cells 11 days postconception and their expression between days 11 and 16 of gestation is confined to the fetal liver. Ia antigen synthesis by fetal liver cells is detectable at day 14. Ia seems to precede Ig as a surface marker of embryonic liver cells, since Ig cannot be detected until day 16 of gestation. H-2 antigens may be immunoprecipitated from day 10 whole embryo cells. F9 primitive teratocarcinoma cells are Ia negative and H-2 negative.
A Newly Described Activity of Guinea Pig IgG1 Antibodies: Transfer of Cutaneous Basophil Reactions
Authors: Haynes J. David , Rosenstein Robert W. , Askenase Philip W.
Published: December 2022

Hapten-specific delayed time course skin reactions containing predominant accumulations of basophils and eosinophils were elicited in newborn guinea pigs after i.v. transfer of small amounts of oxazolone immune serum. The immune serum was fractionated by column chromatography procedures, and the fractions were examined for their ability in transferring this form of cutaneous basophil hypersensitivity (CBH). Only the 7S IgG-containing peak from Sephadex G-200 columns, and only the IgG1-containing fractions from DEAE columns, transferred CBH. An affinity column of bound oxazolone removed the activity from immune serum, and it could be recovered from the column by eluting with soluble oxazolone. About 35 µg of purified IgG1 anti-oxazolone antibody could systemically transfer CBH reactivity. An immunoadsorbant column of anti-IgG1 removed this activity, but a column of anti-IgG2 did not. None of the procedures were able to separate activity in transferring CBH from passive cutaneous anaphylactic (PCA) activity classically associated with guinea pig IgG1 antibody. IgG1 from 8-day immune and 31-day hyperimmune donors were both effective. The average association constant of 8-day antibody was 8 × 10-4 M-1. Transfer of cutaneous basophil reactions can be mediated by low affinity serum 7S IgG1 antibody.
DAN Synthesis and Proliferation of Human Lymphocytes in Vitro
Authors: Bernheim Jan L. , Dorian Randel E. , Mendelsohn John
Published: December 2022

The response of human lymphocytes to phytohemagglutinin (PHA) stimulation was assessed in detailed kinetic studies, in order to define conditions permitting and regulating proliferation. By utilizing live and dead cell counts, cell cycle characterizations, and calculations of rates of entry into S phase and mitosis, it was demonstrated that dilute culture conditions (2 × 105 cells/ml) enable lymphocytes to proliferate for a period of 5 or 6 days. Although cell division occurs in concentrated cultures (2 × 106 cells/ml), net proliferation is not demonstrable due to extensive cell death and earlier decay in the proliferative response. It is projected that without cell death the cell count would rise more than 6-fold in a dilute culture and 2-fold in a concentrated culture under these growth conditions. The kinetic analyses did not demonstrate separate PHA dose thresholds for blastogenesis and entry into S phase. High doses of PHA stimulated a suboptimal response by causing excessive cell death without reducing initial entry into the cell cycle. These results provide a kinetic explanation for the frequent observation of little net proliferation despite substantial 3H-thymidine incorporation in PHA-stimulated lymphocyte cultures. Cell death is identified as a parameter that must be considered in the interpretation of in vitro lymphocyte stimulation studies, and methods for quantifying the role of cell death are demonstrated.
Age-Dependent Loss in New Zealand Mice of Morphological and Functional Characteristics of Thymic Epithelial Cells
Authors: Gershwin M. Eric , Ikeda Richard M. , Kruse Walter L. , Wilson Floyd , Shifrine Moshe , Spangler William
Published: December 2022

The growth and immunologic characteristics of in vitro cultured thymic epithelial cells from NZB, NZW, NZB/W, BALB/c, DBA/2, and N:NIH(S) mice were studied. Cultures were prepared from irradiated (400 R) mice to prevent in vitro autosensitization of thymocytes by epithelial elements, thereby permitting selective growth without the attendant hazards of local cytotoxicity; epithelial cells were enriched and fibroblasts were removed by treatment with a trypsin-EDTA mixture. Forty-eight hours after initial plating, BALB/c, DBA/2, and N:NIH(S) mice all demonstrated a proliferation of large PAS-positive and PAS-negative epithelial cells in a mosaic like arrangement and within 30 days of culture greater than a 95% enrichment of such epithelial cells was achieved. In contrast, proliferation of epithelial cells from thymic explants of New Zealand mice disclosed a marked age dependent relationship. In particular, explants of NZB thymus, age 1 to 52 weeks, failed to produce growth of any cellular elements except fibroblasts. There was, however, evidence of epithelial cell proliferation of NZB/W thymic explants derived from mice less than 8 weeks of age. Nonetheless, serial observations of clones of epithelial cells from NZB/W mice demonstrated vacuolization and degeneration within 2 to 8 weeks of culture, suggesting a pre-programmed defect. Similarly, whereas isolated thymic epithelial cells from BALB/c or young NZB/W mice induced an elevation of thy 1.2 on spleen cells and an increase in responsiveness of nude spleen cells to the T cell mitogen, Con A, cultures from NZB or older NZB/W mice were ineffective. Furthermore, incubation of 7-month-old NZB spleen cells, but not lymph node cells, with a BALB/c thymic epithelial cell monolayer significantly restored the ability to respond to Con A. Moreover, the abnormal proliferation, measured by DNA synthetic response, of NZB spleen seeking but not lymph node seeking thymocytes injected into lethally irradiated C57BL/6 mice was suppressed by preincubation with thymic epithelial cells. Finally, thymic epithelial cells enhanced the ability of NZB thymocytes to suppress a graft versus host reaction. We suggest that functional alterations of thymic epithelial cells in New Zealand mice may be critical to the primary development of autoimmunity and that several of the lymphoid cell defects in these mice, including suppressor cell function, can be corrected by thymic epithelial cells.
Genetic Control of Tolerance Induction to Human γ-Globulin in Mice
Authors: Azar Miguel M. , Ranges Gerald E. , Yunis Edmund J. , Clark Connie
Published: December 2022

Susceptibility to tolerance induction with monomeric human γ-globulin (HGG) was tested in different inbred strains of mice. The results indicated a differential tolerance susceptibility among the strains and that the basis for the variation is genetic in nature. By using a protocol that permits genetic analysis, F1, F2, and backcross generations of the parental strains SJL/J and C3H/Bi were examined. A multigenic control model by H-2-linked and non-H-2-linked genes showing Mendelian autosomal inheritance is proposed.
A Glycoprotein Set Specifically Associated with the Surface and Cytosol of Human Breast Carcinoma Cells
Authors: Leung Julia P. , Plow Edward F. , Nakamura Robert M. , Edgington Thomas S.
Published: December 2022

A new glycoprotein set, specifically associated with human breast carcinoma cells, has been identified, purified to homogeneity, and characterized. This glycoprotein set was first identified in the perchloric acid-soluble glycoprotein fraction of human ductal carcinoma of the breast after removal of known glycoproteins by affinity chromatography. Prototype antiserum to this glycoprotein, after absorption with normal tissues, reacted by gel diffusion with a single constituent of breast tumors but not with other benign or malignant tissues. This antiserum was used to isolate two types of soluble mammary tumor-associated glycoprotein (MTGP). Both types of MTGP appeared to have m.w. of about 19,500 and possess common antigenic determinants. In addition, a firmly integrated insoluble form of MTGP was identified. The soluble types of MTGP differed one from the other in respect to: a) isoelectric point, b) sedimentation velocity, c) carbohydrate composition, and d) buoyant density in cesium chloride. MTGP was found in all six metastatic breast tumors and was independent from plasma and tissue glycoproteins. Antisera to purified MTGP of Types I and II both gave reactions of identity one with the other and with both types of aqueous soluble MTGP. Each antiserum reacted only with extracts of breast tumors and not with any other tissue or type of tumor. MTGP was localized by immunohistochemical methods to both the surface and the cytoplasm of breast ductal carcinoma cells but not normal ductal cells. Positive reactions were also observed in immunologic assays with some spontaneous murine mammary carcinomas but not with the mouse mammary tumor virus. MTGP appears to provide a discrete and characterized new marker for breast carcinoma cells and thus a candidate tumor-specific antigen.
Cellular Basis for a Hapten-Specific State of Tolerance Induced in Vitro
Authors: Kim Young T. , Mazer Theodore , Weksler Marc E. , Siskind Gregory W.
Published: December 2022

A hapten-specific unresponsive state was induced in vitro by the incubation of normal murine spleen cells with highly conjugated dinitrophenylated bovine γ-globulin (DNP-BGG) or a dinitrophenylated copolymer of D-glutamic acid and D-lysine (DNP-D-GL) for 24 hr. After this incubation period spleen cells were washed and cultured for 4 days with the thymic-independent antigen dinitrophenylated polyacrylamide beads (DNP-PAA) or the thymic-dependent antigen trinitrophenylated burro erythrocytes (TNP-BRBC).

Preincubation with either DNP-BGG or DNP-D-GL led to a specific depression of the in vitro anti-hapten plaque-forming cell response. The degree of depression was dependent upon the concentration of the tolerogen and the duration of preincubation. The response to DNP-PAA or TNP-PAA beads was depressed to a greater degree than was the response to TNP-BRBC. The cellular basis of the immunologic unresponsiveness induced by DNP-BGG was attributable to an inhibition of B cell function whereas the unresponsive state induced with DNP-D-GL was due to both a specific inhibition of B cell function and the activation of antigen-specific suppressor T cells.
Characterization of an IgE Receptor Isolated from Cultured B-Type Lymphoblastoid Cells
Authors: Meinke Geraldine C. , Magro Albert M. , Lawrence David A. , Spiegelberg Hans L.
Published: December 2022

For the purpose of characterizing an IgE receptor found on a subpopulation of B lymphocytes, cultured Wil-2WT cells, which bind IgE, were labeled either biosynthetically with 14C-leucine or enzymatically with 125I by the lactoperoxidase method. The cells were lysed with NP-40, and the cleared supernatant was passed over IgE Sepharose-4B immunoadsorbent columns. The material eluted from the column was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Two radioactively labeled proteins having extrapolated m.w. of 86,000 and 47,000 were consistently found in eluates from the IgE columns. A 23,000 dalton band, which may have been a degradation product of the other bands, was often also observed. Reduction of the proteins did not change their m.w., suggesting that they represented single polypeptide chains. Because of their lability, the proteins could only be recovered when three enzyme inhibitors were added to the cell lysates. The eluate from the IgE immunoadsorbent column, incorporated into complete Freund's adjuvant, was injected into a goat. The resulting antiserum, after absorption with cultured lymphoid cells that do not bind IgE, specifically inhibited IgE rosette formation on Wil-2WT cells as well as on other lines of cultured lymphoblastoid cells, chronic lymphatic leukemia, and tonsil and spleen lymphocytes that bind IgE. After absorption with IgE, the antiserum did not cause histamine release from normal or allergic donors' basophils. When the F(ab′)2 fragments of the antiserum coupled to Sepharose-4B were used to isolate proteins from radiolabeled Wil-2WT lysates, both the reduced and unreduced isolated material again showed three components of 86,000, 47,000 and 23,000 m.w. These proteins were not found in lysates of Raji cells, which do not bind IgE. The results suggest that these three proteins are derived from the IgE receptor either as subunits or degradation products and that the IgE Fc receptor on lymphoid cells is antigenically different from that on basophils and mast cells.
H-2 Control of Tolerance Induction to L-Glutamic Acid, L-Lysine-Containing Polymers
Authors: Cheung Nai-Kong V. , Heghinian Krikor M. , Dorf Martin E. , Benacerraf Baruj
Published: December 2022

Previous studies have shown that the copolymer of glutamic acid and lysine (GL) is nonimmunogenic in all inbred mouse strains. In contrast, the immune responses to the GL-containing terpolymers, poly- (Glu, Lys, Phe), poly-(Glu, Lys, Pro), and poly-(Glu, Lys, Ala), referred to as GLø, GLPro and GLA, respectively, are under H-2-linked immune response gene control. The immune defect in nonresponder mice to GL or GL-containing terpolymers can be bypassed by immunization with covalent conjugates of these antigens on the immunogenic carrier, fowl γ-globulin (FγG). Furthermore, injection of nonresponder mice with 1 to 100 µg of GL-containing polymers specifically decreases their ability to develop anti-GL specific IgM and IgG plaque-forming cell responses to a challenge with polymer-FγG conjugates. This tolerance is antigen specific, since the PFC response to the carrier molecules (FγG) is not affected. Tolerance can be induced as long as 15 days before or as late as 5 days after challenge with the polymer-carrier conjugate. Tolerance can be demonstrated in the early phase of the PFC response and cannot be transferred into normal recipients. In contrast, under identical experimental conditions most GLø responder mouse strains cannot be tolerized by GLø, however, some exceptions were noted. Genes associated with the H-2 complex control this tolerance process. The data suggest that an inverse relationship exists between the amount of T cell helper activity and the ease of tolerization.
Liposomes As Model Membrane Systems for Immune Attack
Authors: Ozato Keiko , Ziegler H. Kirk , Henney Christopher S.
Published: December 2022

In attempts to utilize antibody-bound liposomes as model targets for human K cell-mediated cytolysis, a system was developed by which liposomes could be immobilized to plastic surfaces. Phosphatidyl choline liposomes containing 10% (w/w) DNP-caproylphosphatidylethanolamine and labeled with 3H in the lipid phase and 86Rb+ in the aqueous phase were immobilized by incubation with anti-DNP antibody. Two observations indicated that antibody-immobilized liposomes displayed functional Fc portions of antibody: i) they were susceptible to complement-mediated lysis, and ii) they were capable of selectively depleting K cells from human lymphocyte populations.

When lymphocytes were incubated on immobilized liposomes, incremental 86Rb+ was observed, indicating liposome disruption. Such destruction did not, however, correlate with the K cell activity of the lymphocyte populations as measured concurrently with antibody-coated Chang cells. Thus, mouse thymocytes and K cell-depleted human lymphocytes both caused 86Rb+ release from liposomes equivalent to that seen with unfractionated lymphocytes, but only the unfractionated lymphocytes displayed K cell activity. Cell-mediated 86Rb+ release from liposomes could also be distinguished mechanistically from K cell-mediated lysis, for the former occurred in the absence of free divalent cations, whereas the latter did not. These findings underline the complexities in development of model membrane systems as analogues of cell-mediated cytotoxicity.