bluuebunny/crossref_metadata_2025 huggingface dataset:
Isolation of Recombinant IgG Molecules Carrying Heavy Chain Allotypes Transmitted in Repulsion
Authors: Tosi Roberto M. , Tosi Simonetta Landucci , Chersi Alberto
Published: December 2022

Recombinant IgG molecules possessing the heavy chain markers transmitted in repulsion were isolated from a rabbit serum and analyzed. The rabbit genotype was a1/a2; e14/e15, a1 being in coupling with e14 and a2 with e15. Recombinant molecules carrying either a1 and e15 or a2 and e14 were obtained in a relatively pure form by sequential absorption and elution with cross-linked antisera. a1–e15 molecules also were compared by immunoelectrophoresis with other IgG fractions, “nonrecombinant” IgG and “a-minus” IgG, isolated from the same serum. All fractions showed similar immunoelectrophoretic patterns. a1–e15 recombinant molecules were found to have a normal κ:λ light chain ratio and a normal κ-chain allotypes ratio. Furthermore, they could be dissociated into heavy and light chains which elute after gel filtration at the same positions as nonrecombinant molecules.
Immunochemical Studies on Dextrans
Authors: Outschoorn Ingrid M. , Ashwell Gilbert , Gruezo Flavio , Kabat Elvin A.
Published: December 2022

Rabbits were immunized with a hapten-protein conjugate corresponding to a terminal nonreducing chain of five α(1 → 6) linked glucose units plus an additional α-linkage to a six carbon open sugar chain. The antigen was prepared by oxidizing isomaltohexaose to isomaltohexaonic acid and coupling it to bovine serum albumin (BSA). Antisera cross-reacted with dextrans, the amount of dextran required for maximum precipitation being related to its content of α(1 → 6) linkages. This cross-reaction was independent of anti-BSA. Inhibition studies with the isomaltose oligosaccharides from the tri-(IM3) to hepta-(IM7) or octa-(IM8) saccharide with the six antisera showed that four had combining sites complementary to IM6 and two to IM5. For the four sera IM6, IM7, and IM8 were equally effective as inhibitors on a molar basis, better than IM5, IM4 and very much better than IM3. For the remaining two sera IM5, IM6, and IM7 were equivalent and gave better inhibition than IM4 and IM3. Inhibition studies using the aldonic acids series from IM4-COOH to IM7-COOH showed five sera to be maximally inhibited by the pentaonic acid (IM5-COOH) which corresponded to the inhibition obtained with IM6 to IM8 in three sera. The two sera that were maximally inhibited by IM5 showed better inhibition with IM5-COOH and higher members of this series. The last serum showed unique behavior, better inhibition being obtained with the hexasaccharide than with any of the acids. Inhibition by IM4-COOH was equivalent either to IM4, to IM3, or was intermediate for all sera. In one case the IM5-COOH was equivalent to IM4-COOH and IM4. Thus heterogeneous populations of antibodies were formed in each serum as indicated by differences in the relative ease with which inhibition with large and small oligosaccharides and their corresponding acids was obtained.
Induction of Tolerance to an Intracellular Protozoan (Toxoplasma Gondii) by Passively Administered Antibody
Authors: Araujo Fausto G. , Remington Jack S.
Published: December 2022

The humoral antibody response to infection with the intracellular protozoan Toxoplasma gondii was suppressed in adult mice that had been injected with antiserum formed against the same parasite. A small proportion of the mice developed a state of tolerance to Toxoplasma, demonstrable by the absence of Toxoplasma antibody in their sera although Toxoplasma cysts were present in their brains. The tolerant state lasted for 60 days in six mice; between 60 and 90 days after infection, antibody was detected in five of them and in one, the tolerance lasted for 90 days. By day 120 of infection all animals had antibody levels similar to controls.
The Termination of Immunologic Unresponsiveness to Bsa in Rabbits by Transfer of Normal Sibling Thymocytes
Authors: Benjamin D. C.
Published: December 2022

The unresponsive state to bovine serum albumin (BSA) in rabbits was successfully terminated by adoptive transfer of normal sibling thymocytes. These rabbits eventually (13 weeks after transfer) produced normal levels of anti-BSA with normal affinity for BSA. In contrast, BSA unresponsive rabbits that received thymocytes from unresponsive siblings did not produce detectable antibody. The possibility of nonspecific stimulation of antibody formation or the co-transfer of B cells was eliminated. These results are discussed in relation to possible mechanisms for the maintenance of the unresponsive state.
Binding of β-Galactosidase by Cells from Germfree Colostrum-Deprived Piglets
Authors: Decker Janet M. , Kim Yoon Berm , Miller Alexander
Published: December 2022

Cells from the lymphoid organs of germfree colostrum-deprived Minnesota miniature piglets were examined for their ability to bind Escherichia coli β-galactosidase (GZ), with a histochemical detection system. From 0.3 to 1.4% of the cells in the various organs bound GZ; the binding occurred rapidly in the cold with fixed cells, and was specific as shown by competition experiments with high levels of GZ, hemocyanin, and ferritin. Since there is no maternal antibody transfer in pigs, and since the piglets were kept free of antigens and of antibody present in the colostrum, it appears from these findings that cells arise spontaneously before antigen contact that are capable of binding antigen. A proportion of the cells has been shown to bind more than one antigen; this binding is noncompetitive, and therefore suggests that the cells bear more than one specificity of receptor.
The Role of Immunoglobulin in the Neutralization of Trachoma Infectivity
Authors: Barenfanger Joan , MacDonald A. Bruce
Published: December 2022

Eye secretions and plasma from children with active trachoma have been shown to neutralize trachoma organisms. Various preparations of eye secretions were incubated with trachoma organisms and then assayed for infectivity by ocular infection in owl monkeys. Eye secretions with anti-trachoma antibody or immunoglobulin isolated from these eye secretions neutralized infective organisms. Negative controls (eye secretions with the immunoglobulin removed, normal eye secretions, or buffer) did not neutralize trachoma organisms. We conclude that the immunoglobulin fraction is responsible for this neutralization and that the neutralization is type-specific.
Decline and Spontaneous Recovery of the Monoclonal Response to Phosphorylcholine during Repeated Immunization
Authors: Lee William , Köhler Heinz
Published: December 2022

Repeated immunization with T independent phosphorylcholine hapten-carrier conjugates produces progressively unresponsiveness to phosphorylcholine. Only one clone is found to respond and other clones fail to respond throughout the course of immunization. After a rest period the previously unresponsive animals are able to mount a normal response. This response is produced by the same clone which responded during the initial phase of the immunization regimen. A mechanism is proposed by which auto-anti-receptor antibody would protect a repeatedly stimulated responding clone from exhaustive clonal senescence.
Development and Characteristics of In-Vitro Correlates of Cellular Immunity to Rubella Virus in the Systemic and Mucosal Sites in Guinea Pigs
Authors: Morag A. , Beutner K. R. , Morag B. , Ogra P. L.
Published: December 2022

The development of rubella-specific cell-mediated immunity (CMI) in splenic and bronchial lymphoid cells and antibody activity in serum and bronchial washings was studied in guinea pigs after subcutaneous or intranasal inoculation with live attenuated HPV-77 DE/5, RA27/3 rubella vaccines or live Brown strain of rubella virus. The techniques of hemagglutination inhibition, in vitro lymphocyte transformation, and assay of migration inhibitory factor with rubella virus as the antigen were employed to determine antibody and CMI in the systemic and secretory sites. Subcutaneous immunization resulted in regular appearance of serum antibody response. However, no antibody was detected in bronchial washings. Intranasal administration of the virus preparation resulted in little or no antibody response in the serum or respiratory tract. The CMI response after subcutaneous administration of rubella virus was characterized by the specific induction of lymphocyte transformation and migration inhibitory factor activity in the spleen cells without any detectable response in the bronchial lymphoid cells. Intranasal immunization resulted in the appearance of migration inhibitory factor activity in the bronchial cells with little or no response in the spleen cells. The CMI responses were initially detected 2 weeks after immunization. Peak responses were often found at 4 weeks, and no specific migration inhibitory factor or lymphocyte transformation activity was detected after 6 weeks. The results of this study suggest the development of local cellular immune response in the respiratory tract after local application of rubella virus.
Regulation of Homocytotropic Antibody Formation in the Rat
Authors: Taniguchi Masaru , Tada Tomio
Published: December 2022

An antigen-specific subcellular component that augments anti-hapten IgE class homocytotropic antibody (HTA) response was extracted from thymocytes and spleen cells of rats hyperimmunized with dinitrophenylated Ascaris suum extract (DNP-As) or with Ascaris protein (As) alone. This soluble component, when injected into neonatally thymectomized rats, restored the ability to produce anti-DNP HTA and hemagglutinating antibodies of the recipients that were otherwise incapable of producing antibodies upon immunization with DNP-As. The thymocyte extract from either normal rats or those hyperimmunized with DNP-bovine serum albumin (DNP-BSA) did not show the restoring ability for both HTA and hemagglutinating antibodies. Furthermore, this “helper” activity was largely absorbed by the carrier protein, As, but not with hapten, DNP-BSA, indicating the helper component possesses specificity and affinity for the carrier molecule.

Although these antigen-specific characteristics of the helper component are much like those of the previously reported “inhibitory” component of thymus-derived lymphocytes that specifically suppresses the ongoing HTA response, a number of differences in their immunochemical and physicochemical properties were observed. Unlike the inhibitory component, the helper component possessed Fab and µ-chain determinants as revealed by absorption studies. The molecular weight of the helper component was between 200,000 and 100,000 daltons, the value much larger than that obtained for the inhibitory component. These results indicate that IgT-like molecule is involved in the induction of in vivo HTA response, and further suggest that HTA formation is regulated by two distinct antigen-specific subcellular components of thymus-derived lymphocytes in the rat.
Dissociation of T Cell Helper Function and Delayed Hypersensitivity
Authors: Silver John , Benacerraf Baruj
Published: December 2022

The effect of tolerization with varying doses of deaggregated protein carrier on two T cell functions—delayed hypersensitivity (DHS) to the carrier and helper functions for the secondary anti-hapten antibody response to a hapten carrier conjugate—was studied. It was found that high doses of tolerogen inhibit the development of both DHS and helper function, while low doses of tolerogen inhibit the development of helper function but not of DHS.
Peyer's Patch Lymphocytes: Demonstration of the Integrity of Afferent and Efferent T-Cell Functions in the Guinea Pig and Rat
Authors: Levin Douglas M. , Rosenstreich David L. , Wahl Sharon M. , Reynolds Herbert Y.
Published: December 2022

Guinea pig Peyer's patch cells respond to allogeneic cells in vitro in either the one-way or two-way mixed leukocyte reaction. Guinea pig and rat Peyer's patch cells produce a graft vs host (GVH) reaction in appropriate recipients. Guinea pig Peyer's patch cells liberate a lymphokine lymphocyte-derived chemotactic factor (LDCF) when stimulated by the T-cell mitogen concanavalin A. The presence of both the mixed leukocyte reaction and the GVH support the integrity of the afferent limb of T-cell immunity, since both measure the ability of T cells to recognize and respond to cell surface alloantigens. Both the GVH and the LDCF release to concanavalin A demonstrate that the efferent limb of C-cell immunity is also functional.

These studies reinforce the concept that Peyer's patch lymphocytes have the same immunologic potential as those in other peripheral lymphoid compartments.
The Antibody Response to Myelin Basic Protein in Lewis and Fischer Rats as Followed by a Sodium Sulfate Method of Radioimmunoassay
Authors: Day Eugene D. , Wexler Marjorie A.
Published: December 2022

It was found that the sodium sulfate method of radioimmunoassay of 125I-labeled myelin basic protein could be used in the measurement and linear regression analysis of myelin-basic protein-binding activity in Lewis and Fischer rats, and could be adapted to the day-to-day single point measurements of antibody in microliter quantities of serum without untoward loss of statistical precision.
The Distribution of a Rabbit Thymic Antigen and Membrane Immunoglobulins in Lymphoid Tissue, with Special Reference to Mucosal Lymphocytes
Authors: Rudzik O. , Clancy R. L. , Perey D. Y. E. , Bienenstock J. , Singal D. P.
Published: December 2022

We have examined rabbit lymphocytes from several tissue sources for membrane immunoglobulins with antisera specific for α, µ and γ heavy chains, as well as with an antiserum specific for a rabbit thymus lymphocyte antigen (RTLA). Among lymphocytes from bronchus-associated lymphoid tissue (BALT), Peyer's patches, thoracic duct, and gut mucosa exclusive of Peyer's patches, α and µ predominated with roughly equal percentages of each being found. The distribution of cells with γ receptors was approximately half that of either of the two other classes. In splenic lymphocytes, µ predominated, followed, respectively, in numbers by γ and α. Detection of RTLA on almost all thymus lymphocytes (94%), most thoracic duct lymphocytes (72%), and lower numbers of peripheral blood lymphocytes (44%), Peyer's patch lymphocytes (17%), and splenic lymphocytes (20%) indicated a similar distribution of this antigen between the lymphoid organs as has been found for the mouse thymus lymphocyte-specific antigens. Low numbers of RTLA-bearing lymphocytes were detected in both the BALT (18%) and the gut mucosa (11%). An unexpected finding was the number of “null” cells among the gut mucosa populations, bearing neither thymic nor heavy chain markers. These results in toto support the view that lymphoid aggregates in lung and gut may both contain precursor populations of B cells destined for IgA production.
Catabolism Physical, and Immunologic Properties of Endocytosed Isologous and Heterologous γ-Globulins by Mouse Macrophages
Authors: Cruchaud André , Berney Monika , Balant Luc
Published: December 2022

The catabolism of completely endocytosed isologous and heterologous γ-globulins by mouse macrophages was studied in vitro. Mouse, human, and rabbit 125I-IgG were coupled to mouse, human, or sheep erythrocytes either as antibodies or by covalent binding. They were exposed to macrophages for 1 hr and the non-endocytosed erythrocytes were then removed with a Ficoll gradient centrifugation. Catabolism was evaluated after 2, 5, and 18 hr in culture by measuring the radioactivity released into the culture medium as well as the radioactivity that remained associated with cells. It was found that all γ-globulins were catabolized in a similar fashion, and that the type of carrier erythrocytes (isologous or heterologous) had no influence on catabolism. Some of the material that remained associated with macrophages was on the cell membrane and could be removed by trypsin. Some of the material that was released by macrophages was completely degraded but some was either not degraded or only partially degraded. Sucrose density gradient analysis and SDS-polyacrylamide gel electrophoresis showed that this material had kept some physical properties of native γ-globulins. It was also found with the antigen-binding inhibition test and incubation with erythrocytes that the released material contained molecules carrying Fab determinants and was able to bind specifically to erythrocytic antigens. Taken together, these observations show that γ-globulins phagocytosed in the form of antigen-antibody complexes are only incompletely degraded and that the material associated with the plasma membrane of macrophages or found in the culture medium is a product of cell catabolism.
In Vitro Detection of Immune Responses to Mtv-Induced Mammary Tumors: Qualitative Differences in Response Detected by Time Studies
Authors: Blair Phyllis B. , Lane Mary Ann
Published: December 2022

Both BALB/cfC3H females neonatally infected with mammary tumor virus (MTV) and MTV-free BALB/c females possess spleen cells capable of significant activity against target-MTV-induced mammary tumor cells in in vitro microcytotoxicity tests. The responses of the BALB/cfC3H and BALB/c females are different, however, on the basis of time studies. Further, the reactivity characteristic of virgin BALB/cfC3H females is increased in parous females, and part of this reactivity is subsequently lost in multiparous females bearing large tumors. BALB/cfC3H females possess reactive spleen cells as early as 3 weeks of age, indicating that response to MTV antigens develops soon after birth in these neonatally infected mice.
Differential IgA Repopulation after Transfer of Autologous and Allogeneic Rabbit Peyer's Patch Cells
Authors: Rudzik Oleh , Perey Daniel Y. E. , Bienenstock John
Published: December 2022

The distribution and differentiation of rabbit Peyer's patch cells were studied in lethally x-irradiated autologous and allogeneic recipients, 6 days after i.v. transfer. Immunoglobulin Ig-containing cells were detected in both frozen tissue sections and fixed smears of single cell suspensions of spleens using monospecific anti-α, -µ, and -γ antisera in direct immunofluorescence. The tissues of lethally x-irradiated rabbits which had not received any cells were almost devoid of Ig-containing cells whereas the intestinal lamina propria of both autologous and allogeneic Peyer's patch cell recipients had approximately equal numbers of predominantly IgA-containing cells. In the spleen of allogeneic recipients, many large colonies of IgA-containing cells were seen and these cells made up 4.6 to 16.9% (mean 8%) of all spleen cell suspension. The spleens of autologous recipients contained fewer and smaller colonies of IgA cells which made up only 0.5% of all cells in spleen cell suspensions. The numbers of IgM- and IgG-containing spleen cells were small in all animals, however, recipients of allogeneic Peyer's patch cells had four to five times as many as either non-reconstituted lethally irradiated rabbits or autologous recipients. These data confirm that rabbit Peyer's patches are relatively rich in precursors of IgA-producing cells and suggest that histocompatibility differences may either potentiate differentiation of IgA production or lead to trapping of allogeneic Peyer's patch cells in the spleen. Although these experiments did not elucidate the mechanisms of differentiation of IgA-producing cells, the fact that no differences were seen between numbers of IgA cells in gut lamina propria after either autologous or allogeneic cell transfer suggest that different mechanisms may be responsible for appearance of these cells in gut and spleen.
Specific Isolation and Characterization of Antibody Directed to Binding Site Antigenic Determinants
Authors: Claflin J. Latham , Davie Joseph M.
Published: December 2022

The preparation and specificity of antibodies specific for the ligand-binding site of HOPC 8, a phosphorylcholine (PC)-binding mouse myeloma protein, are described. Antiserum to HOPC 8, prepared in rabbits, was adsorbed with an HOPC 8-Sepharose immunoadsorbent and anti-binding site antibodies were eluted with PC. These antibodies reacted with HOPC 8 but not other myeloma proteins, including those with PC-binding specificity different from HOPC 8. The specificity of this anti-HOPC 8 antibody for the combining site region of HOPC 8 was shown by the fact that 1) the interaction of the anti-HOPC 8 antibody preparation with HOPC 8 was completely blocked by PC and 2) the antibody preparation failed to bind TEPC 15 in which the combining sites had been blocked by covalently bound PC groups. Moreover, these anti-binding site antibodies did not react with isolated heavy or light chains, indicating the requirement for a heavy-light chain interaction. By contrast an idiotypic antiserum to HOPC 8 prepared in A/J mice did bind affinity-labeled TEPC 15 and the reaction with HOPC 8 was only marginally hapten inhibitable. Both of the idiotypic determinants detected by these two antisera were present on anti-PC antibody raised in BALB/c mice.
Modulation of in Vivo Antibody Responses by Cholera Toxin
Authors: Kateley John R. , Kasarov Luka , Friedman Herman
Published: December 2022

Treatment of mice with an exotoxin (0.01 µg to 1.0 µg) purified from Vibrio cholerae culture filtrates markedly influenced the immune response to sheep erythrocytes (SRBC) and Escherichia coli lipopolysaccharide (LPS). Simultaneous administration of the toxin (CT) with antigen resulted in a delayed appearance of antibody plaque-forming cells (PFC) during the first few days after immunization, followed by a marked enhancement of both IgM and IgG PFC. The secondary immune response to SRBC was also similarly affected when CT was given together with a second inoculation of SRBC; i.e., a delay in appearance of hemolytic PFC followed by a markedly enhanced IgM and IgG PFC response. Treatment of mice with cholera toxin 1 to 3 days before SRBC or LPS was immunosuppressive. The effect of CT on the level of splenic cyclic AMP appeared related to the effects on antibody formation.
Antibody-Mediated Cytotoxicity after Measles Virus Infection
Authors: Kibler Ruthann , Meulen Volker Ter
Published: December 2022

Antibody cytotoxicity in the presence of complement to measles virus-infected cells was determined in sera of individuals at various stages of measles virus infection. A measles virus carrier cell line, which possessed viral antigens on the cell membrane, was employed as target cells. The cytotoxic effect was measured by 51Cr release from the measles virus-infected cells. A microtiter method was used to determine the titer of antibody cytotoxicity. Cytotoxic antibodies could be detected in serum specimens from individuals after natural measles virus infection and vaccination. Sera from subacute sclerosing panencephalitis (SSPE) patients had only slightly higher titers than were found in sera obtained from individuals 1 week to 3 months after measles virus infection. Titers were much lower in sera from individuals more than 1 year after measles virus infection. Sera from children inoculated with measles virus vaccine lost their ability to react in hemagglutination inhibition, neutralization (NT) and cytotoxicity (CT) tests after absorption with the hemagglutinin from measles virus. SSPE sera treated in a similar manner retained their activity in NT and CT tests. Cytotoxic antibody was, therefore, shown to be directed against the hemagglutinin and some other antigen of measles virus.
Studies on the Specificity of Autoantibodies Produced in Systemic Graft-vs-Host Disease
Authors: Streilein J. Wayne , Stone Marvin J. , Duncan William R.
Published: December 2022

Anti-erythrocyte antibodies were found in serum and on RBC of F1 hybrid hamsters afflicted with systemic graft-vs-host (GVH) disease. These antibodies were shown to be reactive with antigenic determinants unique to hamster erythrocytes compared to heterologous erythrocytes and to non-erythrocytic hamster tissues. They fulfilled the criteria for true autoantibodies by failing to discriminate strain specificities among the domestically maintained isogenic hamster lines that were tested. It is suggested that the excessive production of these antibodies results from the allogeneic effect by which donor T lymphocytes attack host B lymphocytes and stimulate heretofore quiescent self-reactive clones. Moreover, it is proposed that these autoantibodies can function as enhancing antibodies and subvert continued progression of GVH disease. In this regard, they resemble antibodies directed at allogeneic transplantation antigens that have been shown to attenuate and abort GVH processes.
An Expression for the Calculation of Relative Affinities of Antibody-Ligand Interactions
Authors: Paul William E. , Elfenbein Gerald J.
Published: December 2022

An expression is derived which allows the calculation of Ka without a value for total binding sites of antibody. The expression yields unambiguous Ka values for homogeneous populations of antibody molecules. When heterogeneous populations are used, the value of calculated Ka rises as the fraction of sites bound falls.

This expression should be useful in the determination of relative affinity with data obtained from Farr assays of the binding of anti-hapten antibody to hapten. Since values for total binding sites are unnecessary, this equation should be especially applicable for the study of heterogeneous populations of relatively low affinity antibodies present at low concentrations.
Effective Immunity to Dental Caries: Protection of Gnotobiotic Rats by Local Immunization with Streptococcus Mutans
Authors: McGhee Jerry R. , Michalek Suzanne M. , Webb Jerry , Navia Juan M. , Rahman A. F. R. , Legler Donald W.
Published: December 2022

Gnotobiotic rats injected in the submandibular region with killed, whole Streptococcus mutans cells developed salivary antibodies directed to this microorganism. Increased levels of salivary IgA and inhibition and augmentation of agglutinin titers with anti-rat α-antiglobulin suggested that these antibodies were of the immunoglobulin A class. Furthermore, the rats monoinfected and immunized with homologous organisms always had lower mean caries scores than monoinfected, non-immunized rats. This reduction was evident in carious lesions on the buccal surfaces of molars and in those in sulcal areas. These results suggest that local immunization with whole S. mutans cells stimulates a specific salivary IgA response protective against caries resulting from S. mutans infection.
Mixed Leukocyte Culture Reactivity in Operationally Tolerant Rats: Relationship of Lymphocyte-Mediated Reactivity and Serum-Blocking Activity
Authors: Bernstein I. D. , Hamilton B. L. , Wright P. W. , Burstein R. , Hellstrom K. E.
Published: December 2022

Tolerance to Brown Norway (BN) allografts was induced in Wistar Furth (W/Fu) rats by neonatal inoculation of BN bone marrow cells or mixtures of (W/Fu × BN)F1 hybrid spleen and bone marrow cells. Lymphoid cells from rats in which operational tolerance had been induced (maintenance of BN skin graft for more than 100 days) were studied for mixed leukocyte culture (MLC) reactivity. Peripheral blood lymphocytes from 10 of 25 W/Fu rats operationally tolerant to BN skin grafts showed MLC reactivity when exposed to BN antigens in vitro, implying that some degree of MLC reactivity is compatible with prolonged skin graft survival. These same rats also showed cytotoxic activity to BN fibroblasts in vitro regardless of their MLC status. MLC is quantitatively decreased and possibly qualitatively altered as compared to that of control W/Fu lymphocytes. The decrease was specific for the tolerated antigens. Serum from the tolerant rats inhibited cytotoxic but not normal MLC reactivity of W/Fu cells against BN antigens.
Complement Biosynthesis in Vitro by Rat Hepatoma Cell Strains
Authors: Strunk Robert C. , Tashjian Armen H. , Colten Harvey R.
Published: December 2022

Four separate rat hepatoma strains were examined for their capacity to synthesize complement (C). None of the strains synthesized detectable amounts of the first component (C1), and only one strain (7800C1) produced the fourth component (C4). However, each of the strains synthesized significant amounts of biologically active C2 and C3. Three of the four strains also produced C5 and the natural inhibitor of C1 (C1 INH). Two control rat cell strains (fibroblast and pituitary) did not synthesize any detectable C components. Production of C, studied extensively in 7800C1 and H4, was reversibly inhibited by cycloheximide (2 µg/ml) and [14C] amino acids were incorporated into C2, C3, and C1 INH. As assessed by gel filtration, the elution positions of the C components synthesized by the cells in culture were similar to those of the corresponding proteins in normal rat serum. Hydrocortisone (10-6 to 10-7 M) stimulated the production of C3 by H4, but C2 and C5 production were not affected. These C-producing hepatoma cells may prove useful for studies of the control of C biosynthesis.
Induction of Theta-Positive Lymphocytes and Lymphoblasts in Mouse Bone Marrow by Mitogens
Authors: Cohen J. John , Patterson Connie K.
Published: December 2022

With a sensitive technique for the identification of lymphocytes and lymphoblasts bearing the surface antigens ϑ, TL, and immunoglobulin, the induction of ϑ-positive small lymphocytes in bone marrow could be observed. Dibutyryl cyclic adenosine 3′,5′-monophosphate and thymosin were both effective in inducing precursor cells to express ϑ antigen. Concanavalin A and bacterial lipopolysaccharide were also shown to be inducers. After treatment with concanavalin A but not with lipopolysaccharide, lymphoblasts developed in marrow cultures which were positive for ϑ and TL antigens; when spleen was cultured with concanavalin A, lymphoblasts were ϑ-positive and TL-negative. It is concluded that the cell responsive to concanavalin A (and phytohemagglutinin) in bone marrow is a ϑ-negative precursor (stem) cell, which is induced first to express ϑ and TL antigens, and then to differentiate to a lymphoblast.
Specificity of Basophils and Lymphocytes in Cutaneous Basophil Hypersensitivity
Authors: Dvorak Harold F. , Colvin Robert B. , Churchill Winthrop H.
Published: December 2022

Using a rosetting technique, it was found that the vast majority of basophils circulating in the blood or accumulating in the skin reactions of guinea pigs primed for cutaneous basophil hypersensitivity (CBH) lacked demonstrable specificity for sensitizing antigen, whether sheep erythrocytes, a soluble protein, or tumor cells. By contrast, one-third of cells teased from late skin reactions formed specific rosettes as did nearly 80% of circulating basophils in animals receiving repeated doses of whole sheep blood. Unreactive basophils teased from CBH reactions readily acquired rosetting capacity on exposure to immune serum.

With regard to lymphocyte (and hence reaction) specificity, both CBH and classic delayed hypersensitivity (DH) reactions exhibited a high degree of carrier specificity when dinitrophenyl-conjugates were used. Thus, in the hapten-carrier combinations examined thus far, the antigen skin test requirements for both CBH and DH have been identical and are those required for inducing an active lymphocyte response.

These findings indicate that control mechanisms other than homocytotropic antibodies must be sought to explain the accumulation and behavior of basophils in CBH reactions and, coupled with other data, suggest that lymphocytes and/or their products are likely candidates for this role.
Suppression of Maternal Responsiveness to Paternal Antigens by Maternal Plasma
Authors: Pence Hobart , Petty William M. , Rocklin Ross E.
Published: December 2022

The effect of plasma from multigravida women on the expression of cellular hypersensitivity to paternal antigens was investigated. It was found that autologous, but not homologous, maternal plasma specifically blocked MIF production by maternal lymphocytes using their husbands' cells as an antigenic stimulus. The blocking activity could be absorbed out by repeated incubation with paternal cells. Sephadex G-200 fractionation of maternal plasma showed the blocking activity to be in the IgG fraction. The presence of specific blocking factor(s) in maternal plasma which prevent reactivity to paternal antigens may explain in part the success of the fetus as an allograft.
Competition for Antigen by Cell Populations Having Receptors with the Same Specificity but of Different Idiotype
Authors: Bangasser Susan A. , Kapsalis Andreas A. , Fraker Pamela J. , Nisonoff Alfred
Published: December 2022

All normal A/J mice immunized with keyhole limpet hemocyanin-p-azophenylarsonate (KLH-Ar) produce anti-Ar antibodies, some of which share a cross-reactive idiotype. The present results indicate that the biosynthesis of antibody molecules bearing this idiotype does not occur in mice having an excess of lymphoid cells with receptors for Ar that lack the idiotype. This was shown, first, by introducing into normal, nonirradiated mice lymphoid cells from mice which had been suppressed with respect to production of the idiotype and then immunized with KLH-Ar. The recipients failed to express the idiotype upon immunization. Alternatively, cells from suppressed, mice were transferred into lethally irradiated syngeneic recipients, followed 17 days later by normal cells. All recipients expressed the idiotype upon immunization. If, however, the recipients were challenged with antigen between the two cell transfers, antibodies bearing the idiotype were not produced during subsequent immunization. Arguing against active, cell-mediated suppression was the production of the idiotype by normal cells in the presence of cells from a suppressed animal. However, the possibility of active suppression by hyperimmune, suppressed cells was not ruled out. On the basis of the present data, the simplest interpretation is that cells with anti-Ar receptors from immune, suppressed animals, being present in larger numbers, compete successfully for antigen with nonimmune cells and thus prevent the expression of the idiotype. The system may provide a basis for quantitative studies of competition among cells for a limited supply of antigen, particularly if B cell populations are utilized.
Inactivation of Slow Reacting Substance of Anaphylaxis by Human Eosinophil Arylsulfatase
Authors: Wasserman Stephen I. , Goetzl Edward J. , Austen K. Frank
Published: December 2022

Arylsulfatase preferentially present in the human eosinophil as compared to other leukocytes was isolated by sequential gel filtration and cation exchange chromatography. The apparent molecular weight of 60,000, the preferential cleavage of 4-nitrocatechol sulfate (PNCS) over p-acetyl-benzenesulfonic acid (PABS), inhibition by phosphate ions and pH optimum of 5.7 are characteristics of a type II B arylsulfatase. Eosinophil arylsulfatase inactivated purified human slow reacting substance of anaphylaxis (SRS-A) in a time-dependent reaction with the rate dependent upon the enzyme/substrate ratio. That SRS-A inactivation was the result of intrinsic arylsulfatase activity was indicated by association of PNCS cleavage and SRS-A inactivating activity during chromatography, the similar pH optimum for cleavage of both substrates and the capacity of SRS-A to inhibit PNCS cleavage by arylsulfatase. The finding that eosinophil arylsulfatase inactivates SRS-A suggests that eosinophil ingress into the site of an immediate hypersensitivity reaction in response to ECF-A could represent a regulatory function.
Transformation Antigens on Stimulated Lymphocytes
Authors: Bluming Avrum Z. , Lynch Michael J. , Kavanah Maureen , Khiroya Raman
Published: December 2022

Immunologically mediated regulation of lymphoproliferation requires a self-recognition mechanism. This was sought by measuring the ability of blood lymphocytes to recognize transformed, autologous lymphocytes. Human blood lymphocytes incubated with phytohemagglutinin (PHA) for 72 hr, followed by mitomycin C treatment, induced blast transformation of autologous lymphocytes from all 28 healthy adults tested. Blastogenesis was measured by reactor cell incorporation of 3H-thymidine and was greater at 72 than at 144 hr. The role of PHA itself was assessed in several ways. The supernatant of washed, PHA-transformed lymphocytes did not stimulate normal autologous lymphocytes. Lymphocytes incubated with PHA for 1 hr or 72 hr before mitomycin treatment bound equivalent amounts of 131I-PHA; cells treated for 1 hr did not transform and did not stimulate autologous lymphocytes. By contrast, cells incubated for 72 hr did transform and stimulate autologous lymphocytes. Lytic sonication of PHA-transformed lymphocytes abolished their stimulating capability. An identical result was observed in allogeneic mixed lymphocyte reactions after lytic sonication of the stimulating cells. PHA itself maintained its stimulatory capability after sonication. Although N-acetyl-d-galactosamine (NAGAL) competes with PHA for lymphocyte membrane sites, incubation of reactor lymphocytes with NAGAL did not diminish their response to PHA-transformed autologous lymphocytes. These results strongly suggest the presence of autorecognition determinants on membranes of transformed lymphocytes. The relatively rapid reaction to these determinants is consistent with prior in vivo exposure.
Suppression and Enhancement of the T Cell-Dependent Production of Antibody to SRBC in Vitro by Bacterial Lipopolysaccharide
Authors: Hoffmann Michael K. , Weiss Ofra , Koenig Scott , Hirst John A. , Oettgen Herbert F.
Published: December 2022

LPS induced the production of antibody to sheep red blood cells (SRBC) in cultures of spleen cells from normal and T cell-depleted mice, and addition of SRBC to the cultures enhanced this T cell-independent response very little. By contrast, the T cell-dependent production of antibody to SRBC in vitro was suppressed when lipopolysaccharide (LPS) was added at the time when the spleen cells were cultured. Later addition of LPS to spleen cell cultures caused enhancement of antibody production, but only when LPS had not been added before. Addition of T cells that had been primed with SRBC in vivo did not reverse the LPS-induced suppression of antibody production. The data are interpreted to mean that either B cells are rendered incapable of receiving T cell signals in the presence of LPS or that LPS interferes with the appropriate association of cellular components which cooperate in the immune response to SRBC.
Human C3 and C5: Subunit Structure and Modifications by Trypsin and C4̄2̄-C4̄2̄3̄
Authors: Nilsson Ulf R. , Mandle Robert J. , McConnell-Mapes Julia A.
Published: December 2022

The subunit composition of human C3 and C5 was analyzed. Acrylamide gel electrophoresis of the fully reduced and dissociated proteins disclosed a similar structure, consisting of one α and one β subunit, linked together by one or more disulfide bonds. The approximate molecular weights for the α and β subunits of C3 as well as C5 were 140,000 and 80,000 respectively. caused cleavage solely of C3α, whereas trypsin affected both C3α as well as C3β. A characteristic subunit modification by both enzymes indicated that C3α constitutes the source of C3a. as well as typsin exclusively affect C5α. C5a therefore appears to originate from the C5α subunit. The mode of primary cleavage by and trypsin differs, giving rise to different forms of C5b. The question is raised if multiple forms of C5a also exist. It appeared from our studies that certain forms of C5b may retain portions of the α subunit, which could potentially release some biologically active split products following secondary cleavage by the appropriate enzyme.
Polymorphonuclear Leukocytes in Antibody-Dependent Cellular Cytotoxicity
Authors: Gale Robert Peter , Zighelboim Jacob
Published: December 2022

Human polymorphonuclear leukocytes (PMN) lyse antibody-coated target cells in an immunologically specific fashion—antibody-dependent cellular cytotoxicity (ADCC). PMN-mediated cytolysis is independent of complement, de novo protein synthesis, and DNA replication. Cytolysis is rapid, detectable at low PMN:target cell ratios, and exceeds lymphocyte-mediated ADCC. The interaction appears to be mediated via an Fc receptor and is inhibited by aggregated γ-globulin. A role for PMN in host tumor cell immunity is discussed.
Acute Destruction of Rat Skin Grafts by Alloantisera
Authors: Jooste Somarie V. , Winn Henry J.
Published: December 2022

Rat skin transplanted to mice or to allogeneic rats can be acultely and severely damaged by injecting the graft recipients with alloimmune serum of appropriate specificity. This susceptibility of rat skin to alloantisera has been obscured in previous studies by the fact that the extent of serum-mediated damage that occurs is decisively influenced by the interval of time between placement of the grafts and the injection of antiserum. Grafts are resistant to damage during the 1st week after transplantation; they rapidly develop sensitivity during the 2nd week, reaching a peak at 14 to 16 days; and during the next few weeks the sensitivity is lost. Grafts surviving beyond 35 to 40 days become totally resistant to antisera. LBNF1 grafts are more resistant than Le or BN grafts to BN anti-Le serum or Le anti-BN serum, respectively. However, combinations of the two sera can act synergistically to destroy LBN skin that has been grafted to immunosuppressed mice.

This form of serum=mediated tissue damage is thought not to play a significant role in skin graft rejection in the usual circumstances of transplantation since cell mediated immunity is well developed before the acquisition by the grafts of sensitivity to humoral agents. Futhermore, antisera from animals that have rejected a single graft have little or no destructive potential, and hyperimmune animals reject skin verys hortly after grafting, often before the grafts have become vascularized and hence before they are sensitivie to antiserum.
Immunologic and Physicochemical Properties of Enhancing Soluble Factors for IgG and IgE Antibody Responses
Authors: Kishmoto Tadamitsu , Ishizaka Kimishige
Published: December 2022

Rabbits were immunized with dinitrophenyl-coupled Ascaris antigen (DNP-Asc) or ragweed antigen (DNP-Rag) included in aluminum hydroxide gel and their mesenteric lymph node cells were cultured for 24 hr in vitro in the presence of free homologous carrier. The cell-free supernatant thus obtained enhanced both IgG and IgE antihapten antibody responses of DNP-primed cells to DNP-heterologous carrier conjugate (DNP-keyhole limpet hemocyanin). Since the cell-free supernatant obtained from Rag-specific cells enhanced antibody response of hapten-primed cells raised by immunization with DNP-Asc, no carrier specificity was involved in the enhancement. It was found that treatment of primed cells with 10-5 M pactamycin suppressed the formation of the enhancing soluble factor, whereas the factor was readily formed in the presence of 2 µg/mol of cytosine arabinoside in the culture. The results indicated that cell proliferation was not required but de novo synthesis of protein was essential for the formation of soluble factors(s). The enhancing factor was not absorbed by either carrier-coated or anti-carrier antibody-coated immunosorbent. It was also found that the enhancing factor was formed by incubating primed cells with carrier-coated Sepharose. The cell-free supernatant containing no free carrier enhanced both IgG and IgE anti-hapten antibody responses. The activities of the cell-free supernatant to enhance IgG and IgE antibody responses were not absorbed by anti- Fab, anti-γ- or anti-µ-chain antibody immunosorbent, indicating that the nonspecific enhancing factor did not possess immunoglobulin determinant. The cell-free supernatant was fractionated by sucrose density gradient ultracentrifugation and by gel filtration with three radiolabeled proteins, i.e., IgG, ovalbumin, and cytochrome C as markers. Enhancing activity for IgG antibody response was recovered in a fraction between ovalbumin peak (40,000 m.w.) and cytochrome C peak (20,000 m.w.). The activity for IgE antibody response was recovered in a fraction containing IgG marker (150,000 m.w.). By block electrophoresis, both activities were detected in β globulin fraction. The results suggested that different T cell factors are involved in the IgG and IgE antibody responses.
Characterization of Immunoglobulin-Bearing and Other Small Lymphocytes in Mouse Bone Marrow by Sedimentation and Electrophoresis
Authors: Osmond D. G. , Miller R. G. , von Boehmer H.
Published: December 2022

125I-antiglobulin binding and radioautography have been used to define the sedimentation and electrophoretic properties of immunoglobulin-bearing and other small lymphocytes in bone marrow.

Bone marrow cell suspensions from CBA mice were exposed to 125I-labeled rabbit anti-mouse immunoglobulin for 30 min at 0°C. Cell fractions were collected after either sedimentation at unit gravity or continuous free-buffer film preparative electrophoresis. The sedimentation profiles of labeled, antiglobulin-binding small lymphocytes and of unlabeled small lymphocytes were identical, peaking at 2.6 mm/hr, except for 5% of the labeled small lymphocytes which sedimented at 3.8 to 4.1 mm/hr. In electrophoretic fractions most labeled small lymphocytes were contained in a peak of low mobility and unlabeled small lymphocytes were of intermediate mobility, but high mobility fractions also contained small numbers of labeled and unlabeled small lymphocytes. Small lymphocytes comprised 80 to 85% of all nucleated bone marrow cells in fractions taken at the peak of the small lymphocyte distribution profile after either sedimentation or electrophoresis, accompanied mainly by either erythroid or granulocytic precursor cells, respectively.

The results demonstrate that small lymphocytes with readily detectable surface immunoglobulin in the bone marrow closely resemble those in peripheral lymphoid tissues with respect to their sedimentation and electrophoretic properties. The possibility is raised that the rapidly sedimenting immunoglobulin-bearing small lymphocytes in bone marrow are a functionally distinct group of potential precursor cells. Non-antiglobulin-binding small lymphocytes in bone marrow resemble electrophoretically the double negative (immunoglobulin- and θ-negative) small lymphocytes in other lymphoid tissues. Small lymphocytes of high electrophoretic mobility, prominent in the θ- bearing lymphocyte populations of peripheral lymphoid tissues, are few in bone marrow. Lymphocyte-rich suspensions, containing either high or low proportions of immunoglobulin-bearing small lymphocytes, can be separated from bone marrow by electrophoresis.