bluuebunny/crossref_metadata_2025 huggingface dataset:
The Tolerant Cell: Direct Evidence for Receptor Blockade by Tolerogen
Authors: Aldo-Benson Marlene , Borel Yves
Published: December 2022

Autoradiographic studies were done with spleen lymphoid cells of normal and tolerant mice to investigate three questions: 1) Does the specificity of lymphocyte receptors change during tolerance? 2) Are the antigen-binding cells (ABC) eliminated in tolerant animals? 3) Does the tolerogen and/or antigen stay on the surface of the ABC in tolerant animals? BDF1 mice were made tolerant by dinitrophenyl isogeneic IgG (DNP-MGG, the tolerogen) and the state of tolerance was tested by challenging the animals with dinitrophenyl-keyhole limpet hemocyanine (DNP-KLH, the antigen). It was found that the lymphocyte receptor of the ABC was specific for the hapten DNP in normal and tolerant animals. Thus, we have no evidence that the hapten-specific B cell receptor changes its specificity during tolerance and recognizes antigenic determinants present on the tolerogen but not the antigen. The number of splenic lymphocytes with receptors free to bind either the antigen or the tolerogen was the same in normal animals and in animals tolerant for 1 week, whereas it was markedly decreased in animals tolerant for 4 weeks. For determining whether the tolerogen and/or the antigen stayed on the cell surface receptor of the ABC in tolerant animals, indirect autoradiography with labeled anti-DNP antibody was done. Purified antibody to DNP was labeled with 125I and reacted with splenic lymphocytes. We found that animals tolerant for 1 week had significantly more cells which bound anti-DNP than either normal animals or animals which received DNP-KLH. Animals tolerant for 4 weeks had even larger numbers of cells which bound anti-DNP-125I than the mice tolerant for 1 week. Blocking studies showed that the binding of anti-DNP in tolerant animals was specific for DNP on cell surfaces.

These experiments demonstrated the functional divergence in the tolerogenic and immunogenic forms of the same antigenic determinant (DNP). The tolerogen remains on the cell surface of the ABC although the antigen does not. They also show that the DNP clone of cells is not deleted in tolerance. This provides direct evidence for the receptor blockade mechanism of tolerance induction and the existence of the tolerant cell (i.e., ABC with receptor “paralyzed” by tolerogen). We suggest that the existence of this tolerant cell may account for the mechanism of acquired and self tolerance.
The Release of Antigen Molecules from Macrophages: Characterization of the Phenomena
Authors: Calderon Jesus , Unanue Emil R.
Published: December 2022

Macrophages were pulsed with 125I-labeled hemocyanin and then cultured in media devoid of any radioactive material. A great part of the cell-bound 125I was released into the culture fluid in a form not bound to protein; however, a small amount (3 to 7%) was released as protein-bound 125I. The released protein-bound 125I was associated with live, viable macrophages and did not derive from dishes nor from lysis of dead cells. About ¼ of the released protein was immunochemically reactive with anti-keyhole limpet hemocyanin (KLH); the remaining ¾ most likely represents KLH in which the immunoreactive groups lost their original conformation. Trypsin treatment of macrophages did not affect the release of 125I-KLH, suggesting that the product derives from the cell interior. A great majority of the 125I KLH tends to be of smaller size than the original molecule. The release of KLH decreases with time of culture but can be reactivated if fresh medium is added. We conclude that the cell conserves a small pool of intracellular antigen in a form or in a compartment which is slowly catabolized; from this pool a small amount is slowly released into the cell exterior, probably by a process akin to endocytosis but in reverse.
Immunity to Murine Sarcoma Virus-Induced Tumors
Authors: Gorczynski Reginald M.
Published: December 2022

Experiments have been performed to examine the immune status in animals carrying tumors which are in their progressive phase of growth. Unfractionated spleen or lymph node cells of BALB/c mice, inoculated 10 to 14 days previously with murine sarcoma virus (MSV), have no demonstrable immunity in vitro to MSV-viral antigens. On fractionation of the spleen cells from such animals, however, large and small cells can be detected which react in well defined assays for immunity in this system. A cell population of intermediate size is capable of suppressing this activity.

Treatment of the suppressing cell population with anti-θ or anti-immunoglobulin antiserum has provided data to suggest that the suppression is caused by a cell with immunoglobulin moieties on its surface. Moreover, evidence is presented to show that this suppression is nonspecific, in that the induction of protein synthesis in thymus-associated (T) cells in response to the mitogen phytohemagglutinin (PHA) is also suppressed by a cell with the above characteristics. The significance of these findings in relation to the status of growth of the tumor in the host is discussed.
Macrophage Receptors for Migration Inhibitory Factor (MIF), Migration Stimulatory Factor (MSF), and Agglutinating Factor
Authors: Fox Roy A. , Gregory Douglas S. , Feldman Joseph D.
Published: December 2022

The action of macrophage migration inhibitory factor in fetal calf serum (FCS-MIF) was inhibited by l-fucose and by blood group substances and bovine submaxillary mucin, both of which contain l-fucose. FCS-MIF agglutinated human erythrocytes and guinea pig peritoneal exudate macrophages. Agglutination of these cell types was inhibited by l-fucose and N-acetyl-d-galactosamine. Human type O erythrocytes removed MIF from FCS. Supernatants of antigen stimulated guinea pig lymphocytes, containing MIF, agglutinated guinea pig peritoneal exudate macrophages which could be inhibited by l-fucose and N-acetyl-d-galactosamine; and human type A erythrocytes which could be inhibited by N-acetyl-d-galactosamine. A macrophage migration stimulatory factor in fetal calf serum (FCS-MIF) was inhibited by N-acetyl-d-galactosamine. These observations suggest that macrophages and erythrocytes have distinct membrane receptors for MIF and MSF and that these receptors contain monosaccharides.
Relationship of Immunoglobulin to Complement Receptors of Human B Cells
Authors: Abrahamsohn Ises , Nilsson Ulf R. , Abdou Nabih I.
Published: December 2022

By using the disappearance of the surface Ig from human peripheral lymphocytes as a parameter of modulation, at 37°C it was shown that anti-Fab antibody is a more effective and faster modulator of the surface Ig than anti-Fc, anti-IgM, or anti-IgG antibodies. The C-binding site was demonstrated to be fully independent and distinct from the surface Ig and a stable marker of Ig-bearing cells. The C receptor can be blocked by the addition of fresh C5-sufficient, C5-deficient mouse serum, or fresh autologous human serum, at 37°C before binding to EAC by a mechanism still to be defined. Both IgM and IgG cells possess the C receptor. The relationship of Ig to C receptors and their role in cell activation by the specific antigen is discussed.
Ontogeny of Immunoglobulin-Bearing Lymphocytes and DNP-Specific Antigen-Binding Cells in Guinea Pigs
Authors: Davie Joseph M. , Paul William E. , Asofsky Richard , Warren Robert W.
Published: December 2022

The ontogeny of immunoglobulin (Ig)-bearing and antigen-binding lymphocytes in guinea pigs was examined. At 54 days of gestation, fetal spleen contained adult percentages of IgM-bearing lymphocytes, but few which bore either IgG1 or IgG2, immunoglobulin. By term, the fraction of cells bearing each of the major Ig classes had reached, or was approaching, adult levels. In addition, the appearance of Ig-bearing cells with receptors specific for 2,4-dinitrophenyl (DNP) groups was determined. By 50 days of gestation considerable numbers of these cells were found. Virtually all these cells possessed surface Ig of the µ class. Only at term did IgG2-bearing DNP-binding cells appear. In the adult guinea pig, the majority of DNP-specific lymphocytes possess surface Ig of the IgG2 class.
Immunogenesis from Cultured Marrow and Thymus Cells
Authors: Schmiege Susan K. , Miller Harold C.
Published: December 2022

Bone marrow cells and thymocytes were cultured together in a system that provided a suitable microenvironment for maturation of the various lymphoid cell types. The thymus-derived cells collaborated with marrow-derived cells to yield a biphasic antibody response that was not unlike primary and secondary in vivo responses. The kinetics of thymocyte-antigen reactivity and the maturation into functional T lymphocytes indicated rapid cell division over the first 9 days in vitro. These T cells were capable of serving as educated helper cells in induction of B cell terminal differentiation into antibody-forming cells. In addition these cells supported proliferation of NDV virus in the Bloom et al. assay which serves as a corollate of delayed hypersensitivity.
A Simple Method for the Isolation of the Subcomponents of the First Component of Complement by Affinity Chromatography
Authors: Assimeh Seth N. , Bing David H. , Painter Robert H.
Published: December 2022

The three subcomponents of the first component of human complement, C1q, C1r and C1s have been separated and purified by a simple method with affinity chromatography on a column of chemically modified agarose to which human IgG has been covalently linked. This method employs human serum as a starting material, and separates active C1 from serum without the need for euglobulin precipitation. Large quantities of hemolytically active and physicochemically and immunochemically homogeneous C1s can be obtained by a subsequent preparative polyacrylamide electrophoresis step. C1q and C1r may be purified by DEAE-cellulose chromatography followed by gel filtration on Sephadex G-200. A fourth protein which was recovered from the affinity column appears to be a subcomponent of the C1 complex.
Active and Inactive States of Immunologic Unresponsiveness
Authors: Scibienski R. J. , Harris L. M. , Fong S. , Benjamini E.
Published: December 2022

Immunologic tolerance was induced in newborn mice by administration of 1, 5, 10, or 20 mg of lysozyme. These mice were subsequently challenged with a mixture of lysozyme and bovine serum albumin (BSA), or with a covalently coupled conjugate of lysozyme and succinylated BSA. When challenged at 6 weeks of age none of these mice responded to lysozyme when administered admixed with BSA. Of the mice challenged with the lysozyme-SBSA conjugate, those tolerized with 1 or 5 mg of lysozyme responded with antilysozyme antibodies but those tolerized with 10 or 20 mg of lysozyme did not. When challenged at 10 weeks of age, mice which had been tolerized with 20 mg of lysozyme mounted a low level antilysozyme response to both the mixture and the conjugate. Assessment of the response of these animals to BSA and SBSA revealed that the anti-SBSA responses of the conjugate-challenged animals were significantly suppressed whereas the anti-BSA response of the mixture-challenged animals was unaffected.

Tolerance was also induced in adult mice, by the injection of 20, 50, or 100 mg of lysozyme. When challenged 12 days after cessation of the tolerizing regimen, all of these mice exhibited suppressed antilysozyme responses regardless of the challenge antigen, although the antilysozyme responses to the conjugate were slightly higher than to the mixture. No suppression of either anti-BSA or anti-SBSA responsiveness was seen in any of these adult tolerant animals.

It was concluded that administration of low doses of tolerogen to neonatal mice results in only T cell tolerance whereas administration of higher doses of tolerogen also effects B cell responsiveness. Furthermore, neonatally induced tolerance appeared to involve some form of active suppression which did not appear to be a factor in the adult tolerized mice. This suppression was only demonstrable at the B cell level.
Leukemic Serum Inhibition of Phytohemagglutinin-Induced Transformation
Authors: Humphrey G. Bennett , Lankford Janice , Oleinick Samuel , Nitschke Ruprecht
Published: December 2022

The influence of leukemic serum on phytohemagglutinin-P (PHA-P)-induced lymphocyte transformation was studied. Twenty of 22 pretreatment sera collected at diagnosis inhibited the transformation of normal lymphocytes stimulated with PHA-P by more than 50%. In these experiments with PHA-P, kinetic studies of transformation demonstrated that leukemic serum inhibited rather than delayed transformation. The inhibition of transformation by PHA-P was assumed to be due to binding of PHA by components of leukemic serum because the inhibition could be overcome by an increase in the PHA concentration. A biologic assay of the PHA content of lysates of lymphocytes preincubated with PHA and leukemic serum further substantiated the PHA-binding mechanism of leukemic serum. Whether the differences observed in the binding capacity of normal and leukemic serum are qualitative or quantitative remains to be determined.
Increase in Gross (G) Antigen Sites on the Surface of AKR Virus-Induced Rat Lymphoma Cells after Treatment with Trypsin
Authors: Brandchaft Phyllis B. , Boone Charles W.
Published: December 2022

G (Gross) leukemia virus-associated surface antigens (G antigens) on AKR virus-induced Fischer rat lymphoma cells were measured with radioiodine-labeled syngeneic antiserum. Treatment of the lymphoma cells with trypsin resulted in a 3-fold increase in the number of surface antigen sites as compared to EDTA-treated cells. An increase in sites was seen after treatment with as low as 0.025 mg/ml trypsin. Prolonged treatment with trypsin did not decrease the number of G surface antigens, although all of the sites could be removed from the cells by treatment with papain. No increase in the number of antigen sites was observed after neuraminidase treatment of the cells. Blocking experiments with C57BL/6 anti-K36 (G-typing) mouse serum confirmed that the majority of the sites remaining on the cells after trypsin treatment were G cell surface antigens (GCSA). G antigens were also released from the cells by treatment with either trypsin or papain. Released G antigens were sensitive to degradation by trypsin.
Affinity of Murine and Human Interferon for Concanavalin A
Authors: Besancon Françoise , Bourgeade Marie-Françoise
Published: December 2022

Mouse and human white blood cell interferon are specifically complexed to concanavalin A covalently linked to Sepharose 4B beads. This reaction does not take place when α-methyl-D-mannoside is added prior and simultaneously to interferon. After fixation, murine interferon can be displaced from concanavalin A by this sugar. Similarly, in a sensitive tissue culture system, contact with cell membranes detaches the active interferon molecule which, after diffusion, induces the antiviral state in cells located at distant sites from the beads. The Sepharose-concanavalin A interferon complex is more stable at 4°C than uncomplexed interferon.
Correction of T Cell Function by Thymosin in New Zealand Mice
Authors: Gershwin M. Eric , Ahmed Aftab , Steinberg Alfred D. , Thurman Gary B. , Goldstein Allan L.
Published: December 2022

There has been a great deal of interest in the preparation and purification of thymosin, a soluble product of calf thymus (1). This material is believed to be a thymic hormone and like other similar thymic factors described (2) is capable of restoring skin allograft rejection and ability to induce graft-vs-host (GVH)5 disease in neonatally thymectomized mice (3, 4). Further thymosin stimulates lymphoid tissue growth (4), and induces the appearance of T cell markers in bone marrow cells (5). There have been many suggestions that thymosin is critical in preventing autoimmune disease in humans. Indeed levels of thymic hormones are reported low in systemic lupus erythematosus (SLE), and are stated to decrease with age (6).

New Zealand NZB × NZW F1 (NZB/W) female mice, after 7 months of age, manifest a large number of immunologic abnormalities including decreased ability to reject skin allografts, decline in capacity of lymphoid cells to respond to allogeneic cells or other mitogens, and diminished primary response to sheep red blood cells (SRBC).
Enhancement of Immunogenicity of a BALB/c Myeloma Protein in BALB/c Mice
Authors: Fraker Pamela J. , Cicurel Lucienne , Nisonoff Alfred
Published: December 2022

Antiidiotypic antibodies to the BALB/c myeloma protein, LPC-1 (IgG2a(κ)) were induced in BALB/c mice by immunization with protein LPC-1 copolymerized with glutaraldehyde to either rabbit IgG or, preferably, C57BL IgG. The copolymers were much more immunogenic than monomeric LPC-1 or polymerized LPC-1. Tests for antibody activity were carried out by using Fab fragments of monomeric LPC-1 as ligand. The use of copolymers as immunogens should facilitate the study of the rejection of myeloma tumors by isologous mice actively producing antiidiotypic antibodies.

When C57BL mice, which differ from BALB/c with respect to heavy chain allotype, were used as recipients, antiidiotypic antibodies were elicited by monomeric LPC-1 as well as by polymers. In CBA mice, which are of the same allotype as BALB/c, monomeric LPC-1 was ineffective but a homopolymer of LPC-1 was an effective immunogen. Since the homopolymer was a very weak immunogen in the BALB/c strain a factor other than known allotypic determinants must play a role. The mechanism of enhancement of immunogenicity by polymerization is discussed.
Hapten-Specific IgE Antibody Responses in Mice
Authors: Hamaoka Toshiyuki , Newburger Peter E. , Katz David H. , Benacerraf Baruj
Published: December 2022

The present study was undertaken to carry out a detailed comparative analysis of the parameters most conducive to generation in inbred mice of antibody responses of the IgG vs IgE class, and more specifically, helper T cell function participating in antibody responses of these two classes. The data presented in this paper demonstrate the complex factors, i.e., the immunogenic strength, dose of antigen, and the nature of adjuvant vehicle employed for immunization, which determine the magnitude of helper function generated for either one or the other (or both) antibody classes. Although immunization regimens may be defined which result in some dissociation of IgE and IgG antibody responses, the overall impact of the evidence obtained indicates that the existence of distinct T cells for the regulation of these responses will be most difficult to demonstrate directly in the context of the numerous factors contributing to the generation of helper functions. Moreover, such a conclusion must be viewed with considerable caution since the data can equally be explained by differences in sensitivity of IgE and IgG B cells to the effects of the same activated T cells.
Lymphoid Tissue Architecture
Authors: Friedberg S. H. , Weissman I. L.
Published: December 2022

The morphogenesis of the neonatal spleen, mesenteric node, and Peyer's patches has been characterized in terms of the distribution of T antigen-positive and immunoglobulin-positive cells by using immunofluorescent markers. Newborn BALB/c mouse nodes have many T, but few Ig-bearing cells; conversely, the spleen at this stage contains only rare T cells, while prominent accumulations of Ig-positive cells occur surrounding conspicuous vessels. At about 3 days of age, T cells appear in increasing numbers in a perivascular position, displacing the cuffs of Ig-positive cells, and thus the basic pattern of organization charateristic of the adult splenic white pulp is established. The mesenteric node and Peyer's patches, in the early neonatal period, are both characterized by the presence of scattered Ig-bearing cells, in contrast to the highly organized distribution in the spleen. By the 3rd or 4th day, however, the Ig-positive cells of the nodes and patches form accumulations distributed in roughly the adult pattern. The paucity of T cells in the spleen and of Ig-bearing cells in the mesenteric node of the newborn has been shown not to be due to their failure specifically to trap infused adult T and B cells.

Kinetic analysis of Ig-positive cell accumulation reveals that such cells in the neonatal mesenteric node are probably not generated in situ. Moreover, the level of proliferation in the Peyer's patches is insufficient to account for the increase in even the splenic Ig-bearing cells, and it is thus highly unlikely, at least in the neonatal mouse, that the Peyer's patches are analogs of the avian Bursa of Fabricius.
Specific Suppression of Delayed Hypersensitivity: The Possible Presence of a Suppressor B Cell in the Regulation of Delayed Hypersensitivity
Authors: Neta Ruth , Salvin S. B.
Published: December 2022

Delayed hypersensitivity to purified protein antigens in incomplete Freund's adjuvant is of a transient nature in guinea pigs. This decrease in delayed reactivity can be transferred adoptively to a sensitized recipient with lymphoid cells from the spleens and peritoneal exudate of donors 8 days after sensitization with ovalbumin. When such mixtures of donor lymphoid cells are adoptively transferred at subsequent times after sensitization, the delayed hypersensitivity of the recipient is not suppressed. When a hapten-protein conjugate, such as para-aminobenzoic acid-azo-hen egg albumin is used as a sensitizing antigen in the donor, and when the donor cells are removed at various time intervals after the decline in delayed skin tests to the carrier protein as antigen, such as 10 to 23 days after sensitization, adoptive transfer of lymphoid cells into sensitized recipients suppresses the elicitation of delayed skin hypersensitivity to the carrier. When spleen cells are compared with peritoneal exudate (PE) cells in their suppressive capacities, only the spleen cells show suppressive action. The duration of delayed responses varied inversely with the dose of the immunosuppressive drug in guinea pigs treated with varying doses of cyclophosphamide 72 hr before sensitization. In view of the observations that PE cells are an enriched source of T cells and that the particular schedule of treatment with cyclophosphamide affects rapidly dividing cells, the suggestion is made that a suppressor B cell may regulate the activity of a T cell in the expression of delayed skin hypersensitivity.
Modulation of Polymorphonuclear Leukocyte-Mediated Antibody-Dependent Cellular Cytotoxicity
Authors: Gale Robert P. , Zighelboim Jacob
Published: December 2022

Human polymorphonuclear leukocytes (PMN) are able to mediate the immunologically specific cytolysis of antibody-coated target cells—antibody-dependent cellular cytotoxicity (ADCC). The initial interaction is divalent cation dependent and is inhibited by ethylenediaminetetraacetic acid (EDTA). Agents capable of elevating adenosine cyclic 3′,5′ monophosphate (cyclic 3′,5′ AMP) and exogenous cyclic 3′,5′ AMP are inhibitory to PMN-mediated ADCC activity, while cholinergic agents capable of elevating guanosine 3′,5′ cyclic monophosphate (cyclic 3′,5′ GMP) and exogenous cyclic 3′,5′ GMP exhibit a facilitory effect. Colchicine in concentrations inhibitory to microtubule formation and pharmacologic agents capable of interfering with granule release are inhibitory to PMN ADCC activity.
Anti-Idiotypic Antibody in Rat Transplantation Immunity
Authors: McKearn T. J. , Stuart F. P. , Fitch F. W.
Published: December 2022

Lewis × Brown Norway (LBN) F1 antiserum to an idiotype of Lewis anti-BN alloantibody (referred to as the Id-1 idiotype) was used to test a variety of Lewis alloimmune sera for the presence of Id-1. Several such Lewis sera raised by BN antigen injection at monthly intervals possessed the Id-1 idiotype. Hyperimmune Lewis anti-BN alloantiserum raised by six weekly injections of BN antigen, however, did not possess the Id-1 idiotype, but appeared to possess anti-idiotypic antibody to Id-1 instead. The anti-BN hemagglutinin titer, anti-idiotypic antibody titer to Id-1, and levels of the Id-1 idiotype were then measured in individual weekly serum samples from Lewis rats repeatedly immunized with BN antigen. The anti-BN hemagglutinin titers followed a cyclical pattern, with the Id-1 idiotype predominating only in the first cycle of the anti-BN hemagglutinin response. Anti-idiotypic antibody was demonstrable in five of eight repeatedly immunized rats, and in each of these five animals the appearance of anti-idiotypic antibody to Id-1 followed the clearance of Id-1 from the serum. Anti-idiotypic antibody may play a role in the regulation or control of an established immune response.
The Plaque Cytogram Assay
Authors: Thornthwaite Jerry T. , Leif Robert C.
Published: December 2022

A method, the plaque cytogram assay, for preparing fixed, stained dispersions of plaque-forming cells (PFC) and rosette-forming cells (RFC) is described. Both light and scanning electron microscopy (SEM) studies demonstrate that these cells are pleomorphic. Eight types of PFC and seven types of RFC can be simultaneously identified by light microscopy in which small and medium lymphocytes are the predominate cell types of the two immunocompetent cell classes. SEM shows fairly smooth cell surfaces on the PFC, while the RFC have a rough surface. Enzymatic staining of PFC and RFC for lipase demonstrates that none of the PFC are stained positive for lipase, while approximately 27% of the RFC are lipase positive. Finally, several distinct types of plaques and rosettes are shown in the plaque cytogram assay.
In Vitro and in Vivo Cytotoxicity to EL-4 Sarcoma in New Zealand Mice
Authors: Gershwin M. Eric , Klingenstein R. James , Chused Thomas M. , Steinberg Alfred D.
Published: December 2022

The immune response to EL-4 tumor was studied in female NZB/W, BALB/c and CDF1 mice after both in vivo and in vitro immunization with spleen cells as attackers and 51Cr-labeled EL-4 cells as targets. NZB/W mice demonstrated marked changes with age in the cytotoxic response after both in vivo and in vitro sensitization, whereas the other strains did not. These changes were characterized by hyporesponsiveness at 9 months of age, and hyper-responsiveness at 6 months of age, relative to 1 month of age. Antibody to EL-4 tumor was found in unimmunized NZB/W mice and tended to increase with age. Nevertheless, the cytotoxic antibody response in NZB/W mice after in vivo sensitization was similar at 1 month and 6 months of age, but was reduced at 9 months of age. Nine-month-old NZB/W mice failed to mount an adequate cytotoxic response and were killed by the tumor, whereas 1- and 6-month-old animals mounted cytotoxic responses and survived.
Cellular Immunity to Leishmania Donovani
Authors: Skov Clare B. , Twohy Donald W.
Published: December 2022

C57BL/6J mice were subjected to various combinations of immunosuppressive treatments and infected i.v. with amastigotes of Leishmania donovani. In normal mice the parasite population reached its peak about 24 days post-infection, after which parasite numbers gradually declined but they were still detectable on day 60. The course of infection was not altered in mice which had been thymectomized, lethally irradiated, and reconstituted with syngeneic marrow cells. Moderate lethally irradiated, and reconstituted with syngeneic marrow cells. Moderate suppression of acquired resistance resulted when thymectomized-irradiated mice were reconstituted with marrow cells which were previously treated with anti-brain associated θ serum and/or when recipient mice were treated with azathioprine on the same day they were infected. Splenectomy alone or together with irradiation failed to suppress immunity, but a long-lasting high level of immunosuppression resulted when splenectomy was combined with all the above treatments. These results suggest that 1) small numbers of thymus-dependent cells which survive irradiation and thymectomy or which may be present in the bone marrow may be sufficient to mount an immune response to L. donovani in mice, and 2) that T cells play an important role in acquired resistance to L. donovani in mice.
A Pathologic IgM Occurring in Serum as Pentamer, Dimer, and Monomer
Authors: Parr Dorothy M. , Connell George E. , Powell Andrew J. , Pruzanski Waldemar
Published: December 2022

IgM was recovered from the serum of a patient (Den) with Waldenstrom's macroglobulinemia by preparative electrophoresis and gel filtration. In the latter step three species were resolved, with sedimentation coefficients of 7.7S, 11.5S, and 19.0S. Molecular weight determinations showed that the three species were monomeric, dimeric, and pentameric IgM, respectively. The 19S and 11S species were not dissociated in 6M urea. Serologic, immunologic, and biochemical studies indicated that the constituent light and heavy chains of each species were the same. Light chains prepared from 19S IgM (Den) contained a second component identified as J chain. Light chains from 11S and 7S IgM (Den) did not contain this component.
Differentiation of Lymphoid Cells: Enhancement of the Induction of Immunoglobulin Production by Thymus Cells and a Product Secreted by Thymus Cells
Authors: Okumura Ko , Shinohara Nobukata , Kern Milton
Published: December 2022

The induction of immunoglobulin M production, as a consequence of the incubation of rabbit spleen cells for 72 hr in cell culture in the absence of added antigen, was markedly enhanced when thymus cells were cultured along with spleen cells. In contrast, immunoglobulin production by lymph node cells was only slightly stimulated by the presence of thymus cells during culture. Immunoglobulin production by spleen cells was equally enhanced, regardless of whether the thymus cells were derived from unimmunized animals or from animals immunized with a dinitrophenyl-antigen. Thymus cells added to cultures of spleen cells at 48 hr were still markedly stimulatory even though the induction process in the absence of thymus cells appears to be largely complete at 48 hr. Ficoll density gradient separation of spleen cells revealed some fractions that were slightly enhanced and others that were markedly enhanced by thymus cells. Finally, a cellfree mediator secreted into the culture medium by thymus cells also enhanced immunoglobulin production to about the same extent as thymus cells.
In Vivo and in Vitro Production of a Slow Reacting Substance in the Rat Upon Treatment with Calcium Ionophores
Authors: Bach Michael K. , Brashler John R.
Published: December 2022

Injection of the divalent metal ion-specific ionophores, A 23187 and X 537A, into the peritoneal cavity of the rat resulted in the release of histamine. In addition, there was produced a large amount of a substance which caused slow contractions of the atropinized, mepyramine-treated guinea pig ileum. A similar activity was also produced when mixed peritoneal cells were incubated with the ionophores in vitro in the presence of physiologic buffer solutions containing calcium ions. Production of the activity in vivo was unaffected by pretreatments of the animals to deplete them of mast cells, polymorphonuclear neutrophils, or both, or by the depletion of complement activity with cobra venom factor. The activity was stable to alkali and to incubation with chymotrypsin. Chromatography on XAD-2 resin followed by partition chromatography on silicic acid resulted in the recovery of the activity in an ethanolic ammonia eluate. These chemical characteristics and the yield of the recovered activity are reminiscent of the behavior of slow reacting substance of anaphylaxis (SRS-A) under the same conditions.
Thymus Dependence of the Spontaneous Induction of the in Vitro Anamnestic Antibody Responses to Human Serum Albumin and Keyhole Limpet Hemocyanin
Authors: Stavitsky Abram B. , Tew John G. , Harold Weldon W.
Published: December 2022

Recently we reported that IgG antibody synthesis was spontaneously induced—no extrinsic antigen required—simply by transferring lymphoid cells from a human serum albumin (HSA)3 immunized rabbit into cell culture (1). Initiation of this spontaneous in vitro response was attributed to cell-associated persisting antigen, whose immunogenicity was subject to an antibody feedback mechanism (1). It was proposed that this in vitro phenomenon represented the essential features of the mechanism whereby antibody synthesis is maintained and regulated in vivo for months or years after antigen injection (1). Subsequently, additional evidence was presented that: a) persisting antigen was responsible for induction of the spontaneous antibody response, and b) this persisting antigen was bound to the surface of an accessory cell such as a sessile macrophage or dendritic type cell (2).

The purpose of the present study was to determine whether thymus-derived (T) lymphocytes were required for the induction of the spontaneous antibody synthesis.
Immunogenic Properties of Modified Antigen E
Authors: Ishizaka Kimishige , Okudaira Hirokazu , King Te Piao
Published: December 2022

Ragweed antigen E loses its major antigenic determinant after denaturation in 8 M urea. The urea-denatured (UD) antigen and α-polypeptide chain isolated from the denatured molecule possess their own antigenic determinant(s) but lack the major determinant of the native molecule. The UD antigen and α-chain, however, are capable of priming mouse T cells specific for antigen E. Priming of A/J mice with the modified antigen enhanced both IgG and IgE antibody responses to antigen E. Both UD antigen-primed spleen cells and α-chain-primed spleen cells collaborate with DNP-primed cells to give an adoptive secondary anti-DNP antibody response to DNP-ragweed antigen in syngeneic irradiated mice. Pretreatment of A/J mice with an i.v. injection of α-chain partially suppressed both IgE and IgG antibody responses to antigen E. Weekly injections of α-chain or UD antigen to antigen E-primed animals depressed on-going IgE antibody response, and suppressed secondary IgE antibody response to antigen E. Transfer of spleen cells from animals treated with the modified antigen to irradiated recipients followed by challenge with native antigen showed that the adoptive secondary response was suppressed by injections of modified antigen to the donors. The results indicate that the immunocompetent cell population was changed by the treatment and provide an experimental model to analyze the immunologic effect of hyposensitization treatment.
Host-Derived Antibody-Forming Cells in Lethally Irradiated Mice
Authors: Pilarski Linda M. , Cunningham Alastair J.
Published: December 2022

The technique of adoptively transferring an immune response by injecting donor cells into irradiated recipient mice is based on the assumption that the host is rendered immunologically inert by irradiation. We present evidence that these mice are not inert; in the presence of donor cells of various types they mount a substantial immune response against both heterologous erythrocytes and dinitrophenylated bovine serum albumin. This restoration of the host requires donor cells for its expression and can account for up to 100% of the total response. The problems associated with this finding are discussed.
Spontaneous and Methylcholanthrene-Enhanced Thyroiditis in Buf Rats
Authors: Silverman David A. , Rose Noel R.
Published: December 2022

Virgin female Buffalo strain (BUF) rats that were treated with methylcholanthrene (MC) had a higher incidence of lymphocytic thyroiditis than untreated BUF rats. The degree of infiltration was also greater in MC-treated rats. The incidence of thyroiditis in retired breeder female BUF rats approached that found in MC-treated rats. The susceptibility to the effect of MC was not expressed in most F1 hybrids of BUF and Lewis (LEW), a strain of rat that does not develop thyroiditis after treatment with MC.
E Receptors on Blasts from Untreated Acute Lymphocytic Leukemia (All): Comparison of Temperature Dependence of E Rosettes Formed by Normal and Leukemic Lymphoid Cells
Authors: Borella L. , Sen L.
Published: December 2022

The presence of receptors for sheep erythrocytes (E) and surface Ig on the bone marrow blasts was investigated in 29 children with untreated acute lymphocytic leukemia (ALL). In 6 of them more than 50% of the bone marrow blasts formed E rosettes, while in none of the 29 were surface Ig detected. The six children with rosette-forming blasts had WBC τ; 5 × 104/mm3 at admission and 4 of them presented with thymic enlargement. E-positive ALL blasts and normal human thymocytes formed rosettes after incubation for 1 hr at 4°C or 37°C. In contrast, normal human peripheral T lymphocytes from blood, spleen, and pleural fluid also formed rosettes at 4°C but these rosettes dissociated at 37°C. In two patients with E-positive ALL, the disappearance from blood of cells forming rosettes at 37°C during the 1st week of treatment paralleled the reduction in circulating blasts. Conversely, after 6 days of therapy almost one-half of the remaining cells formed rosettes at 4°C and had the morphologic features of normal lymphocytes. We conclude that, in contrast to peripheral T cells, normal thymic cells and E-positive blasts share the property of forming E rosettes after 1-hr incubation at 37°C. In patients with E-positive ALL this property may be used to evaluate drug effects upon leukemic and normal T lymphocytes.
Inhibition of Proliferation of Lymphoma Cells and T Lymphocytes by Suppressor Cells from Spleens of Tumor-Bearing Mice
Authors: Kirchner Holger , Muchmore Andrew V. , Chused Thomas M. , Holden Howard T. , Herberman Ronald B.
Published: December 2022

We have recently demonstrated suppressor cells in spleens of mice bearing tumors induced by Moloney murine sarcoma virus (MSV) which were non-T cells and inhibited phytohemagglutinin Moloney (PHA)-induced DNA synthesis of syngeneic normal spleen cells. From the present study, the suppressor cells appeared to be macrophages since they were radioresistant, inactivated by carrageenan, and removed by adherence columns and an iron/magnet technique. We have also found that suppressor cells were still fully active when added 16 hr after the mitogen, thus indicating that early mitogen-induced changes were not the target of suppressive action. It appeared that suppressor cells inhibited metabolic events related to the initiation of DNA synthesis and that they had a selective effect on proliferation-dependent lymphocyte effector functions. PHA-induced cytotoxic reactivity which in our system is largely independent of DNA synthesis was not depressed but actually enhanced in MSV spleens. Cytotoxicity of MSV spleen cells against syngeneic lymphoma cells was unaffected by suppressor cells whereas lymphocyte stimulation by mitomycin C-treated syngeneic lymphoma cells was inhibited. MSV spleen cells also inhibited DNA synthesis of cultured murine lymphoma cells. This function was only slightly diminished after treatment with anti-θ serum plus guinea pig complement. Furthermore, spleen cells from MSV tumor-bearing nude mice were as effective as spleen cells from their heterozygous littermates, thus suggesting that T lymphocytes are not the main effector cells of inhibition of lymphoma cell DNA synthesis. The inhibitor cells were radioresistant, inactivated by carrageenan, and removed by adherence columns and the iron/magnet technique. These data strongly suggest that the inhibitor cells of lymphoma cell DNA synthesis are macrophages and that they belong to the same group of cells as the suppressor cells of PHA-induced lymphocyte proliferation.
Depression of Humoral Immunity to Sheep Erythrocytes in Vitro by Friend Virus Leukemic Spleen Cells: Induction of Resistance by Statolon
Authors: Weislow Owen S. , Wheelock E. Frederick
Published: December 2022

The in vitro antibody plaque-forming cell (PFC) response of normal spleen cells exposed to sheep erythrocytes in Marbrook chambers was depressed by the addition of Friend virus (FV) leukemic spleen cells. Fewer than 105 leukemic cells inhibited the PFC response of 107 normal cells. This immunodepression could not be produced with sonicated, irradiated, or mitomycin C-treated leukemic cells or with cellfree FV. It could be blocked by FV immune serum but was unaffected by high titers of purified interferon. Interferon did not inhibit the immune response of normal spleen cells to sheep erythrocytes. Spleen cells from mice treated with statolon or chlorite-oxidized oxyamylose statolon but not Newcastle disease virus or poly rI:poly rC were resistant to immunodepression in vitro by FV leukemic spleen cells. The unique ability of statolon to prevent immunodepression by leukemic spleen cells may be the basis of its FV leukemo-suppressive activity in vivo.
Immunopathology of Mouse Hepatitis Virus Type 3 Infection
Authors: Dupuy J. M. , Levey-Leblond E. , Prevost C. Le
Published: December 2022

Normal adult A strain mice are resistant to MHV3 infection. A strain mice immunosuppressed by 600 rads of x-irradiation or by anti-lymphocyte serum treatment became susceptible to the virus and died with specific lesions of the liver and high virus titers. However, mice immunized with MHV3 before sublethal x-irradiation resisted a second injection of virus. Resistant adult (A × C3H) F1 hybrids undergoing graft-vs-host (GVH) reaction became highly susceptible to MHV3 injected 8 days after parental cell injection. Virus titer 3 days after injection was 2 logs higher in mice undergoing GVH than in controls. However F1 hybrid mice resisted virus challenge when the first injection of virus was given 2 weeks before GVH induction. In addition, thymectomy also modified the behavior of resistant animals toward virus infection. It appears, therefore, that cell-mediated immune functions play an important role in resistance of mice to MHV3.
Immunologic and Biochemical Properties of the Histamine-Sensitizing Factor from Bordetella Pertussis
Authors: Lehrer Samuel B. , Vaughan John H. , Tan Eng M.
Published: December 2022

A highly potent extract of the histamine sensitizing factor (HSF) of Bordetella pertussis was isolated by extraction of bacterial cells with urea buffer and subsequent gel filtration. This preparation of HSF also contained leukocytosis-promoting activity and adjuvant activity for reaginic and hemagglutinating antibodies. Digestion of this extract with pronase or trypsin partially destroyed histamine-sensitizing activity, leukocytosis-promoting activity, and adjuvant activity for reaginic antibody, but did not affect adjuvant activity for hemagglutinating antibody. Antisera to HSF was prepared by immunizing rabbits with either whole bacteria or partially purified extract. These antisera contained several precipitating antibodies to Bordetella pertussis extract demonstrated by immunodiffusion and immunoelectrophoresis. Antisera added in vitro to Bordetella pertussis extracts or passively administered in vivo to mice, reduced or abolished all biologic activities except adjuvant activity for hemagglutinating antibody. These results suggest that HSF might be an antigenic component of Bordetella pertussis which also possesses leukocytosis-promoting activity and adjuvant activity for reaginic antibody.
Increased Resistance of Immunoglobulin a Dimers to Proteolytic Degradation after Binding of Secretory Component
Authors: Lindh Erik
Published: December 2022

The contribution of secretory component to the stability of secretory IgA against proteolysis has been studied by a new approach, i.e., by comparing the proteolytic degradation of IgA dimers with the degradation of the complexes formed in vitro between these proteins and secretory component. The results show that attachment of secretory component to the dimeric backbone of the secretory IgA molecule is accompanied by a significantly increased resistance of this backbone against digestion by both trypsin and pepsin. This protective effect may be a physiologic function of secretory component or may be due merely to unspecific blocking by secretory component of one or more sensitive peptide bonds in the IgA backbone.
Immunochemical Quantitation of the Third Component of Guinea Pig Complement in Fluid Phase and Bound to Cell Surfaces
Authors: Ohanian Sarkis H. , Borsos Tibor
Published: December 2022

A method is described for the determination of the absolute amount of guinea pig (GP) C3 bound to cell surfaces. The test is based on the inhibition of anti-C3 by C3. The C3 to be measured is added to a standard amount of anti-C3 antibody and the residual activity is determined by its capacity to sensitize EC43gp for lysis by GPC. By comparing the observed lysis with inhibition-of-lysis curves obtained with known C3 antigen, the amount of test antigen can be determined. The method is simple, rapid, and reproducible and can detect nanogram amounts of C3.

The C3 content of six normal guinea pig sera was found to be 1365 ± 56 µg/ml (ranges 1120 to 1510 µg/ml) by the inhibition method described in this paper.

Guinea pig hepatoma cells, line-1 and line-10, harvested from the peritoneal cavity of guinea pigs contained approximately 6.7 × 103 to 6.2 × 104 GPC3 per cell whereas tumor cells sensitized with anti-Forssman or specific anti-tumor antibody contained from 1 to 9 × 105 C3 per cell. Susceptibility to killing by antibody and GPC could not be correlated with the amount of GPC3 bound to the surfaces of the tumor cells.

Comparisons made between the amounts of C3 bound to antibody-sensitized sheep erythrocytes and nucleated cells suggest there is a fundamental quantitative difference in the fixation of selected C components.
The Properties of Lymphocytes Which Carry Immunologic Memory of φX 174
Authors: McGregor D. D. , Mackaness G. B.
Published: December 2022

Cells that carry immunologic memory of φX 174 have been studied in the rat. Memory cells could not be detected in thoracic duct lymph before the 10th day after priming. Thereafter, they increased rapidly in number to reach a plateau by the 4th week. This long latency was not due to a protracted process of maturation of cells formed and delivered to the lymph during the early post-induction period because memory never appeared in rats immunized adoptively with cells obtained before the 11th day. The fact that φX memory cells are resistant to inhibition by vinblastine also indicates that memory is carried by cells which are released from regionally stimulated lymphoid tissue only after they have ceased dividing and become functionally mature.

Adoptively acquired memory decays rapidly at first, but slowly from the 2nd week after cell transfer. This suggests that two cell populations of differing half-life are involved. If two distinct populations are necessary to the full expression of φX memory, both populations recirculate because the memory cells in lymph draining the sites of the primary response and those reaching the central lymph of adoptive hosts give parallel dose-response curves.