bluuebunny/crossref_metadata_2025 huggingface dataset:
Multiple Myeloma: An Immunologic Profile
Authors: MacKenzie Malcolm R. , Paglieroni Teresa
Published: December 2022

Seventy-four patients with multiple myeloma, 17 untreated and 57 treated, were studied to characterize their peripheral blood lymphocytes. PBL were studied for E, EAC, and EA rosette-forming cells, SIg, and Fc receptorbearing cells. The responses to PHA, Con A, and PWM were assessed as well as their ability to stimulate or to respond in a MLC. Finally, the capacity of mitogen-stimulated lymphocytes to lyse Chang cells, CRBC, and PHA-stimulated lymphoblasts was examined. These results were compared with a group of normals and patients with benign monoclonal gammopathy. In untreated myeloma patients there was a normal percentage of T cells, but an abnormal distribution of B cells as judged by a decrease in SIg-bearing cells, as well as an increase in EAC rosette-forming cells. Subpopulation analysis showed a marked increase in EAC rosetteforming cells without SIg. PHA, Con A, and PWM, and response in MLC were all normal. However, the ability to stimulate in MLC was significantly depressed. Treated myeloma patients had similar findings, except that the response to PWM was significantly depressed. The capacity of PWM-stimulated cells to lyse target cells was depressed in both groups. The results indicate that, in the peripheral blood of myeloma patients, there are populations of lymphocytes characterized by the presence of the EAC receptor without SIg, which are deficient in the capacity to stimulate an MLC response and the ability to be cytotoxic when stimulated by PWM. The results form a baseline for the study of abnormal lymphoid function in human myeloma.
A Spontaneously Occurring Complex of β2-Microglobulin and a Fragment of γ-Chain of IgG: Isolation from the Urine of a Patient with Plasma Cell Leukemia and Characterization
Authors: Seon Ben K. , Pressman David
Published: December 2022

A complex of a fragment of γ-chain of IgG and β2-microglobulin (β2M) was isolated from the urine of a patient with plasma cell leukemia. The approximate m.w. of the γ-fragment was 19,000 and this γ-fragment was found to be associated with β2M noncovalently. The complex could be dissociated in 7.5% (v/v) n-propanol which suggests an important role of hydrophobic bonds in the association of the γ-fragment and β2M. The β2M did not bind monoclonal IgG from this patient. Several anti-β2M antisera tested which had been prepared with free β2M did not react with the β2M associated with the γ-fragment, but reacted with the free β2M obtained by the dissociation of the complex.
Studies on Demyelination by Activated Lymphocytes in the Rabbit Eye
Authors: Stoner Gerald L. , Brosnan Celia Fildes , Wisniewski Henryk M. , Bloom Barry R.
Published: December 2022

The role of products of activated lymphocytes (PAL) in the pathogenesis of primary demyelination has been investigated in the rabbit eye model. Supernatant factors were prepared from rabbit lymph node cells specifically activated in vitro with non-brain antigen (PPD) or nonspecifically activated by mitogen (concanavalin A-Sepharose). The supernatants were shown to contain migration-inhibitory factor (MIF) activity and skin-reactive factor (SRF) activity, mediators of delayed-type hypersensitivity. The active supernatants were injected into the vitreous of the right eye to determine their ability to promote demyelination of the retinal fibers. The left eye received a supernatant prepared from unstimulated cells. In normal rabbits and in rabbits sensitized with lung or kidney, or FCA alone, or infected with living BCG, the lymphocyte products failed to damage myelin directly. Nor was the mononuclear cell infiltrate induced by the PAL specifically destructive to myelin. However, in rabbits sensitized to homologous spinal cord, the lymphocyte products induced a demyelinating lesion within the retinal fibers. This result indicates a role for PAL, in concert with other as yet undefined functions, in the initiation of an autoimmune demyelinating lesion in the CNS. The use of PAL to induce a mononuclear cell infiltration of the rabbit eye extends the usefulness of the rabbit eye model for studies of the pathogenesis of demyelination.
In Vitro Tolerance Induction of Bone Marrow Cells: A Marker for B Cell Maturation
Authors: Metcalf Eleanor S. , Klinman Norman R.
Published: December 2022

B cell subpopulations have been defined by various morphologic criteria and differences in functional capacity. In a recent report from this laboratory, the in vitro splenic focus system was utilized to define a subpopulation of neonatal splenic B cells on the basis of their susceptibility to tolerance induction. Since this susceptibility to tolerance induction progressively decreases with age, tolerance susceptibility in this system has been attributed to the immaturity of the cells which constitute developing B cell clones. In the present study, the splenic focus technique was utilized to characterize the maturational state(s) of neonatal and adult bone marrow B cells as assessed by the susceptibility of these cells to tolerance induction, and to compare these responses to the responses of splenic B cells. The results demonstrate that the majority of neonatal bone marrow B cells reach maturity, by the criterion of tolerance susceptibility, later than splenic B cells. Furthermore, in contrast to the spleen, a portion (25%) of bone marrow B cells remain susceptible to tolerance induction even in the adult. Thus, the generative pool of B cells appears to shift from the spleen to the bone marrow after the first few days of neonatal life and resides primarily in the bone marrow thereafter. Finally, the susceptibility of B cells to tolerance induction may serve as a marker for the functional maturation of any given B cell population, and consequently may help to define distinct B cell subpopulations within the B cell pool.
Receptor-Binding Sites on C3 and C3b
Authors: Dierich Manfred P. , Bokisch Viktor A.
Published: December 2022

Human erythrocytes carry C3b receptors; Daudi lymphoid cells carry exclusively C3d receptors. With these two types of cells, it could be shown that isolated, uncleaved human C3 and soluble C3b possess two stable binding sites: SBS1-specific C3b receptors, and SBS2-specific for C3d receptors. Upon binding of freshly cleaved C3b (nascent C3b) via its labile binding site (generated on C3b through cleavage of C3) to the C3 acceptors on cell surfaces, the SBS2 becomes concealed. Kinetic experiments show that immediately after the action of fetal calf serum or partially purified C3b-inactivator on EAC1423b the SBS2 is accessible again. The reappearance of SBS2 does coincide with cleavage of surface-bound C3b into C3c and C3d but not with the release of C3c from the cell; C3c remains attached and is released only with delay. Concomitant with the cleavage event the number of SBS1 is reduced, stressing the importance of an unaltered steric configuration of the C3b structure for the expression of SBS1. An alternative explanation might be that the SBS1 is located at the site connecting C3c and C3d, so that it becomes altered upon dissection of the two fragments.
Complete Sharing of Light Chain Spectrotypes by Murine IgM and IgG Anti-Streptococcal Antibodies
Authors: Perlmutter Roger M. , Briles David E. , Davie Joseph M.
Published: December 2022

In order to examine the diversity of antibody light chains, we have developed an analytic isoelectric focusing procedure which permits the routine analysis of L chains from antibodies raised in individual mice. We have used this technique to demonstrate that the light chains of IgM and IgG anti-group A streptococcal antibodies raised in SWR mice are probably shared. Interestingly, numerous light chain spectrotypes are shared between individual mice whose 7S antibody focusing patterns differ.
The Mechanism of Activation of the First Component of Complement by a Univalent Hapten-IgG Antibody Complex
Authors: Goers John W. , Ziccardi Robert J. , Schumaker Verne N. , Glovsky M. Michael
Published: December 2022

We have previously shown that the univalent hapten, 2,4-dinitrophenyl (DNP)-polylysine when bound to anti-DNP-IgG antibody, yields a 7S complex which is a powerful activator of the classical pathway (Goers et al., 1975, J. Biol. Chem., 250:4918). In this paper we compare this univalent hapten-antibody complex with unassociated antibody, reduced and alkylated antibody, and IgG to which polylysine has been covalently attached. Of these, only the univalent hapten-antibody complex fixes complement. However, both antibody and reduced and alkylated antibody bind equally well to the C1q subunit of the first component, whereas only the univalent hapten-antibody complex shows enhanced binding to the intact first component, C1. Moreover, the antibody-hapten complex or hapten alone will bind to activate C1r,C1s to give C1̄r,C1̄s. The C1s, by itself, is not activated by hapten. Moreover, reassociated C1, composed of C1q,C1r,C1s, is activated by the hapten-antibody complex but not by hapten alone. These results allow us to postulate the existence of a polylysine-sensitive activation site on C1r,C1s which is concealed when C1r, C1s is associated with C1q. Furthermore, we suggest that the binding of hapten-antibody complex to C1 induces a conformational change which causes this site to become exposed. Finally, since all of the substances tested bind equally well to C1q, but only the hapten-antibody complex activates C1, we suggest that peripheral subunits on C1q serve only as anchors to bind C1 to antibody-coated cell surfaces or immune complexes and are not directly involved in the C1 activation process. Activation may occur, we speculate, when the antibody-hapten complex combines with the central stalk of C1q,C1r,C1s.
Molecular Control of B Cell Triggering by Antigen-Specific T Cell-Derived Helper Factor
Authors: Shiozawa C. , Singh B. , Rubinstein S. , Diener E.
Published: December 2022

Soluble, carrier-specific helper factor extracted from thymus-derived (T) cells primed to the carrier in vivo has been used to analyze the relationship between the degree of T cell help required to stimulate a primary antibody response in vitro and the molecular structure of the antigen. In order to define the conditions for optimal T cell help, we have constructed antigenic probes which consist of a number of serum proteins conjugated to a carbohydrate backbone. When these probes were trinitrophenylated for use as hapten-carrier conjugates to stimulate in vitro anti-TNP immune responses, we were able to define the conditions for optimal T cell help as one at which the carrier antigenic determinants are presented to the bone marrow-derived (B) cell in a closely spaced sequence. We suggest such a carrier focuses helper factor molecules by providing an array of determinants to which they can bind and then interact in a concentrated fashion with a small area of the B cell surface membrane.

When comparing the capacity of B cells to cooperate with helper factor from either syngeneic or allogeneic T cells, it was found that for a successful immune response to occur, both cooperating elements have to be of syngeneic origin. This extends the concept of allogeneic restriction between cooperating B and T cells to the level of interaction of soluble T cell-derived helper factor with B cells.
Myeloma-Specific Antibodies: Studies of Their Properties and Their Relationship to Tumor Immunity
Authors: Frikke Maureen J. , Bridges Sandra H. , Lynch Richard G.
Published: December 2022

The anti-idiotypic antibodies (a-Id315) induced in BALB/c mice by immunization with the dinitrophenyl (DNP)-binding, isologous myeloma protein produced by MOPC-315 were analyzed by double antibody radioimmunoassay to determine time of appearance, levels, heavy chain class, inhibitability with DNP-hapten, and relationship to MOPC-315 transplantation resistance. Following 200 µg of RA-315 per immunization, a-Id315 activity and MOPC-315 transplantation resistance were detected after the second of five weekly immunizations, and both increased after each subsequent immunization. After five weekly immunizations, individual sera from 180 mice were found to bind from 50 to 860 µg of 125I-RA-315 per ml. Approximately 95% of the a-Id315 detected after five weekly immunizations belonged to the IgG1 class, and the remaining 5% was IgG2a. Between 80 and 100% of the a-Id315 binding was inhibited by 3 × 10-3 M 2,4-DNP-L-lysine. Immunization schedules using increasing amounts of immunogen, from 5 to 200 µg, or increasing numbers of immunizations, from 1 to 4 showed that the induction thresholds for a-Id315 activity and the myeloma graft resistance were indistinguishable. When mice immunized with 200 µg of RA-315 weekly for 5 weeks were challenged with numbers of MOPC-315 cells sufficient to overcome transplantation resistance in 50 to 80% of the mice, none of the a-Id315 features analyzed differentiated resistant from nonresistant mice. Passive administration of a-Id315 serum did not influence the growth of MOPC-315 cells in normal syngeneic hosts. These observations suggest that the mechanism of idiotype-specific transplantation resistance observed in a-Id315-producing hosts probably involves more than the a-Id315 antibodies alone.
Antibody-Dependent, Eosinophil-Mediated Damage to 51Cr-Labeled Schistosomula of Schistosoma Mansoni: Effect of Metabolic Inhibitors and Other Agents Which Alter Cell Function
Authors: David John R. , Butterworth Anthony E. , Remold Heinz G. , David Peter H. , Houba Vaclav , Sturrock Robert F.
Published: December 2022

The effect of drugs known to inhibit different metabolic pathways or cell functions on antibody-dependent, eosinophil-mediated damage to schistosomula was determined. Eosinophil-mediated damage was completely inhibited by cytochalasin B, inhibitors of glycolysis, aminophylline, and treatment of cells with diazotized sulfanilic acid and tosyllysyl-chloromethyl ketone. The effects of cytochalasin and 2-deoxyglucose were reversible. On the other hand, eosinophil-mediated damage was unaffected by agents which inhibit DNA replication, protein synthesis, oxidative respiration, prostaglandin synthesis, and microtubule aggregation. The findings suggest that alteration of the cell surface membrane and microfilaments prevents damage by interfering with cell-target interactions, that energy derived from glycolysis is required for cytotoxicity, that cell-associated esterases are probably involved, and that cytotoxicity may be modulated by cyclic nucleotides. Some of the attributes of eosinophils that allow it and not other cells to mediate this reaction are discussed.
Negative Allogeneic Effects in Vitro
Authors: Swain Susan L. , Dutton Richard W.
Published: December 2022

Murine T cells can mediate a potent negative allogeneic effect on the capacity of primed cells to develop the secondary antibody-forming cell response to hapten carrier in vitro. This effect is detected when T cells confront responding cells differing at the major H-2 locus. The allosuppression is relatively sensitive to mitomycin treatment and to irradiation. The T cells responsible for the inhibition of antibody formation appear to express the Ly2 but not the Ly1 alloantigen. The secondary response of spleen cells in culture is quite insensitive to positive allogeneic effects. The usefulness of this model in elucidating the mechanism of allosuppression and the relevance of such effects to studies involving genetic restriction on cell interactions is discussed.
Temporal Regulation of the Immune Response to LDH-C4 by an X-Linked Gene in C3H/HeJ and SJL/J Mice
Authors: Marsh James A. , Wheat Thomas E. , Goldberg Erwin
Published: December 2022

Regulation of the immune response is a complex phenomenon under the control of many genes. This process may be analyzed by comparing the immunogenicity of structurally similar but distinct antigens. Both synthetic immunogens with limited numbers of determinants (1) as well as antigenically complex proteins (2, 3) have been used to study immunoregulatory mechanisms. The LDH iso zymes are particularly interesting because they display a high degree of structural similarity but can be distinguished immunologically.

Vertebrate LDH is a tetrameric enzyme. All tissues contain variable amounts of A (or M) and B (or H) subunits which randomly associate to produce five isozymes. The genetic control of the immune response to heterologous LDH-A and -B subunits has been studied in rabbits (4), rats (5), and inbred mice (6, 7). Antiserum to purified LDH-A4 or LDH-B4 reacts only with the isozymes containing the subunits used in immunization but not with the heterologous homotetramer (8, 9).
Letters to the Editor
Authors: Bigazzi Pierluigi E.
Published: December 2022

Dear Sir: We are flattered by the attention with which Dr. Junger has read not only our latest paper in the J.I., but also the other publications on BUF rat thyroiditis by our group. As you can imagine, we are very well aware of our previous reports on the incidence of thyroiditis in animals at 3 months of age or older. However, even in our recent review on spontaneous animal models of thyroiditis (1), we decided not to present a table such as the one Dr. Junger has compiled from our published data, because of the variability observed in several groups of animals. We have noticed thyroiditis at the age of 4 ± 1 months and this is the basis of our statement that the “disease appears first at the age of 4 months.”

With regard to the breakdown of disease by sex, we have indeed distinguished between sexes (see Table I), but because of the already too large number of tables we did not include these data in our J.I. paper.
Isolation of Receptor-Bearing Plasma Membrane Vesicles from Guinea Pig Macrophages
Authors: Scott Robert E. , Rosenthal Alan S.
Published: December 2022

The plasma membrane serves as an important site for the interaction of macrophages and lymphocytes in antigen recognition and of macrophages and foreign agents in inflammatory responses. Despite the critical role of the macrophage membrane in these reactions, studies on the biochemical and functional characteristics of macrophage membranes which need to be performed have been limited by the lack of suitable methods for the isolation of macrophage plasma membranes that maintain functional and antigenic membrane markers. Recently a new method for membrane isolation has been reported which is based on the observation that a variety of sulfhydryl blocking agents can induce the shedding of plasma membrane vesicles of monolayer cell lines. In this paper we demonstrate that guinea pig peritoneal exudate macrophages can also be induced to shed significant quantities of plasma membrane vesicles by such reagents, in particular with a low m.w. aldehyde, and that these vesicles can be isolated by centrifugation. Vesiculation in macrophages is potentiated by disulfide reducing agents, such as dithiothreitol, and by calcium ions. Maximum vesiculation is observed when well spread macrophage monolayers are incubated in a slightly hypertonic vesiculant at pH 7.0 for 2 hr at 37°C. Brief exposure of macrophages to a vesiculant for intervals as short as 30 min is also effective. Macrophage vesicles shed from the cell surface by this procedure have been found to contain receptors for concanavalin A, wheat germ agglutinin, phytohemagglutinin, and lentil lectin. Vesicles also contain Fc and complement (C3) receptors and form rosettes with thymic lymphocytes. It is suggested that macrophage plasma membrane vesicles isolated by this technique should serve as a valuable new system to study the biochemical characteristics of macrophage plasma membranes and their role in immunologic reactions.
Increased Binding and Killing of Neuraminidase-Galactose Oxidase-Treated Tumor Cells by Normal Macrophages
Authors: Piessens Willy F.
Published: December 2022

The binding of Line 10 hepatoma cells to normal syngeneic guinea pig macrophages is increased when the tumor cells are treated with neuraminidase and galactose oxidase (NAGO) before they are added to the macrophage monolayers. The effect is abolished by exposure of the NAGO-treated tumor cells to sodium borohydride.

Line 1 hepatoma cells treated with NAGO or with sodium periodate are killed to a greater extent than untreated tumor cells. This effect can also be reversed by sodium borohydride. Further, periodate-treated macrophages become cytotoxic for unmodified tumor cells.

These results demonstrate that increased tumor cell killing occurs when artificial contacts (presumably via Schiff bases) are established between normal macrophages and tumor cells. They are consistent with the hypothesis that close cell to cell contact is necessary for macrophage-mediated cytotoxicity.
Inhibition of Complement by Mouse Serum: Selective Inactivation of the Fourth Component of Human Complement
Authors: Mandle Robert J. , McConnell-Mapes Julia A. , Nilsson Ulf R.
Published: December 2022

Bound human C4b on EAC4 is rapidly inactivated in the presence of murine serum reagents. The functional characteristics of this inactivation suggest that it is probably caused by factor(s) homologous to human C3bI-C4bI: inactivation is temperature-dependent and occurs without concomitant consumption of the inactivator(s); loss of hemolytic function is associated with the cleavage of the bound C4b into C4c, which is released, and C4d, which is retained on the cell membrane. Murine serum reagents inhibit bound human C4b far more efficiently than C3b and may therefore be employed to selectively inhibit C4b.
Interactions of C-Reactive Protein with the First Component of Human Complement
Authors: Claus D. R. , Siegel J. , Petras K. , Osmand A. P. , Gewurz H.
Published: December 2022

C-reactive protein (CRP), a trace constituent of serum which generally increases dramatically in concentration during reactions of inflammation and tissue destruction, has recently been shown to inhibit multiple functions of both T cells and platelets. Certain interactions of CRP in the presence of human serum have been shown to result in the efficient depletion of C components C1-5, and in the initiation of C-dependent adherence, phagocytosis, and cytolysis. In the present experiments, CRP was compared with antibody for the ability to react with the first component of human complement. Like IgG, CRP was found to bind purified C1̄ and this binding was markedly enhanced upon chemical aggregation or interaction of CRP with one of its binding specificities, the C-polysaccharide (CPS) of the pneumococcus. Both hemolytically active and native CI bound to CPS-coated erythrocytes (ECPS) sensitized with CRP (ECPS-CRP), and in both instances C1̄ could be recovered by transfer to EAC4; approximately equivalent amounts by weight of CRP and rabbit anti-CPS were required for these effects. The interaction between CRP and C1̄ occurred at the level of C1q, as shown by inhibition of binding and transfer of C1̄ by purified C1q, agglutination of ECPS-CRP by C1q and binding of 125I-C1q to ECPS-CRP. Thus, these results provide further evidence for the similarity of CRP to antibody in the nature of its interaction with the first component of human complement at the level of C1q, and support the concept that CRP may have an important role in the initiation and modulation of reactions of inflammation and host defense.
The Role of Macrophages in the Generation of T Helper Cells
Authors: Erb Peter , Vogt Peter , Meier Barbara , Feldmann Marc
Published: December 2022

The generation of T helper cells in vitro requires macrophages or macrophage-derived factors such as genetically related macrophage factor (GRF) or nonspecific macrophage factor (NMF). However, there is a basic difference of T helper cell induction when using particulate antigens. The present study demonstrates that this difference is based on the activation of two different T cell subsets. GRF activates short-lived ‘T1’ cells which amplify the induction of T2 cells, which are the helper cell precursors. Thus, the genetic restriction of T helper cell induction seen with soluble antigen or GRF lies on the level of macrophage or GRF interaction with T1 cells. NMF (or macrophages) and particulate antigens directly activate the helper cell precursor (T2) indicating no requirement for T1-T2 cooperation. The direct activation of the helper cell precursor with particulate antigens does not require histocompatible macrophages or NMF from histocompatible macrophages.

The present results may explain some of the discrepancies reported in the literature concerning the genetic requirements and specificity of T cell activation.
Immune Suppression with Supraoptimal Doses of Antigen in Contact Sensitivity
Authors: Sy Man-Sun , Miller Stephen D. , Claman Henry N.
Published: December 2022

Immunologic suppression was induced in a mouse model of contact sensitization to DNFB by using supraoptimal doses of antigen. In these studies, in vivo measurement of ear swelling as an indication of immunologic responsiveness correlated well with measurement of in vitro antigen-induced cell proliferation.

This unresponsiveness was specific, since supraoptimal doses of DNFB did not interfere with the development of contact sensitivity to another contactant, oxazolone. The decrease in responsiveness is a form of active suppression, as lymphoid cells from supraoptimally sensitized donors transferred suppression to normal recipients. Furthermore, pretreatment with cyclophosphamide (Cy) reversed the suppression seen in supraoptimally sensitized animals but had no effect on the optimal sensitization regimen.

These results indicate that supraoptimal doses of contactants can activate suppressor cells and that precursors of these cells are sensitive to Cy. Such suppressors regenerate within 7 to 14 days after Cy treatment.

The ability of Cy pretreatment to affect supraoptimal sensitization without affecting optimal sensitization confirms other reports indicating that the observed results of Cy treatment depend critically upon the dose of antigen used.
Immunologic Aspects of Human Colostrum and Milk
Authors: Ogra S. S. , Weintraub D. , Ogra P. L.
Published: December 2022

A group of formula-fed infants were administered a single feed of poliovirus IgA antibody-rich human colostrum 18 to 72 hr after birth. Subsequently, the presence of IgG, IgA, and IgM immunoglobulin and poliovirus antibody activity was determined in serial serum and fecal samples of the neonates. Absorption of IgA immunoglobulin from the colostrum to the circulation was observed in three infants who were fed with colostrum between 18 and 24 hr after birth. Another group of infants of tuberculin-positive mothers who were being breast fed by their own mothers were followed for the development of in vitro correlates of cell-mediated immunity against tuberculin after prolonged breast feeding. Tuberculin-specific proliferative response was observed in the peripheral blood lymphocytes of two neonates after 5 weeks of breast feeding. The responses were undetectable after 12 weeks, although the infants continued to breast feed. No tuberculin reactivity was observed in the cord lymphocytes. These observations suggest uptake of IgA immunoglobulin and components of cellular immunity in the intestine during the immediate neonatal period.
Evidence that Human Leukocyte Inhibitory Factor (LIF) Is an Esterase
Authors: Rocklin Ross E. , Rosenthal Alan S.
Published: December 2022

There are known to be two distinct lymphocyte mediators which inhibit cell migration in vitro, migration inhibitory factor (MIF), which inhibits macrophage migration, and leukocyte inhibitory factor (LIF), which influences the migration of polymorphonuclear leukocytes (PMNL). The present studies were carried out to clarify conflicting reports concerning the sensitivity of MIF activity to a serine esterase inhibitor, diisopropyl phosphofluoridate (DFP). Human MIF and LIF activities were generated by stimulating blood lymphocytes with concanavalin A, filtering the supernatants on Sephadex G-100 gels, and collecting the active fractions of each. MIF and LIF preparations were incubated for 30 min at 37°C with varying concentrations of DFP, dialyzed extensively to remove the inhibitor, and tested on their respective indicator cells. The results showed that LIF activity, but not MIF activity, was irreversibly abolished by DFP (10-3 to 10-5 M). Another irreversible terase inhibitor m-[o-(2-chloro-5-fluorosulfonyl-phenylureido)-phenoxybutoxy] benzamidine, in a dose-dependent fashion, abolished LIF activity. The inactivation of LIF by DFP was concentration, time, pH, and temperature dependent, indicating that DFP served as a substrate for LIF. In addition, LIF preparations (but not MIF) were found to contain hydrolytic activity for certain artificial substrates such as benzoyl arginine ethyl ester and tosyl arginine methyl ester but not glysyl lysyl methyl ester. The response of human monocytes to MIF was not altered by pretreatment with DFP or soybean trypsin inhibitor (STI). In contrast, pretreatment of PMNL with DFP and STI prevented their response to LIF. These studies suggest that LIF, but not MIF, is an esterase.
Pathways of Human B Lymphocyte Differentiation: A Clonal Transition between IgM and IgG Synthesis in Leukemic B Lymphocytes
Authors: Rudders Richard A. , Howard Judith P.
Published: December 2022

Fresh human lymphoma cells from patient Gre were found to have both IgG and IgM associated with only one type of light chain (λ) on the surface membrane. After tissue culture (up to 35 days), enzymatic stripping, reagent absorptions, and ultracentrifugation, similar results were obtained. Double fluorescent labeling studies also demonstrated IgM and IgG on more than 90% of single cells. In experiments designed to study redistribution and subsequent polar capping with double labeling (rhodamine and fluorescein) techniques, we found that 1) anti-IgG or IgM and λ reagents formed mixed polar caps, 2) anti-IgG and IgM formed polar caps which suggested segregation of the two labels, and 3) prior staining with either heavy chain antiserum did not block subsequent staining with the other. We interpret these findings as suggesting the presence of a clonal switch mechanism involving IgM and IgG synthesis operating at the B lymphocyte level, presumably via an alteration in the CH gene. Additionally, when secretory Ig was looked for, we found evidence for intracellular (secretory) λ-chain synthesis without corresponding heavy chain synthesis. This latter observation further emphasizes the divergence of the secretory and membrane Ig synthetic pathways proposed by other investigators.
Strain Variation in BCG-Induced Chronic Pulmonary Inflammation in Mice
Authors: Allen Elizabeth M. , Moore Vernon L. , Stevens James O.
Published: December 2022

C57BL/6 mice (haplotype H-2b) responded in a dose-dependent fashion to killed BCG by marked enlargement of the spleen and lung. Neither CBA nor C3H mice (haplotype H-2k) responded to such treatment. Pulmonary inflammation in responder B6 animals was characterized by a marked chronic interstitial and alveolar granulomatous process, and was accompanied by occasional granulomata, hyperemia, and loss of architecture in the spleen. Inflammation in non-responder CBA and C3H animals was minimal in both the lung and spleen. The response does not appear to be controlled by genes within the major histocompatibility complex, but is associated with a C57 background. B10.BR mice (responder background, H-2k) were responder animals and C3H.SW mice (nonresponder background, H-2b) were nonresponders. In addition, all animals tested with a C57 background were responders even though two of these strains were not H-2b (C57BL/Ks, H-2d and C57Br/cd, H-2k). The resolution of the mechanism of genetic control of this response in mice may provide information relevant to possible genetic control of chronic pulmonary inflammation in man.
Lymphocytes Binding Polyriboadenylic Acid and Synthesizing Antibodies to Nucleic Acids in Autoimmune and Normal Mice
Authors: Sawada Shigemasa , Pillarisetty Rao J. , Michalski Joseph P. , Palmer Donald W. , Talal Norman
Published: December 2022

Splenic lymphocytes from both normal and autoimmune mice bind significant quantities of polyriboadenylic acid (poly rA) when incubated with radiolabeled poly rA for 40 min at 37°C. This poly rA binding is specifically inhibited by an excess of nonradioactive poly rA and by anti-mouse immunoglobulin. Poly rA binding is decreased by exposing spleen cells to Pronase and is restored by subsequent culture for 18 to 72 hr. Poly rA-binding activity is associated more with bone marrow-derived than with thymus-derived lymphocytes. These results suggest the presence of autoantigen-binding lymphocytes in normal as well as autoimmune mice.

Furthermore, spleen cells from normal and autoimmune mice cultured for 72 hr synthesize and secrete antibodies to poly rA and DNA. These antibodies can be recovered from the culture supernatants by a solid immunoadsorbent technique and antibody immunoprecipitation. The synthesis of antibodies to nucleic acids by normal spleen cells suggests that autoreactive lymphocytes may be released from normal immunoregulatory control during in vitro culture conditions.
Isolation of the Cytoplasmic Granules of Guinea Pig Basophilic Leukocytes: Identification of Esterase and Protease Activities
Authors: Dvorak Harold F. , Orenstein Neil S. , Dvorak Ann M. , Hammond M. Elizabeth , Roblin Richard O. , Feder Joseph , Schott Chester F. , Goodwin Jane , Morgan Ellen
Published: December 2022

Procedures were developed for isolating highly purified cytoplasmic granules of basophilic leukocytes from guinea pig peripheral blood. The methods involved disruption of cells in 0.34 M sucrose followed by a series of membrane filtrations and fractionation on sucrose density gradients. These preparations, up to 95% pure basophil granules by electron microscopy, contained a mixture of neutral esterases-proteases including caseinolytic activity; both trypsin- and chymotrypsin-like serine hydrolases were identified by means of appropriate inhibitors. Localization of at least one such activity to the basophil granule was confirmed by a cytochemical method; this activity was absent in contaminating lymphocytes and eosinophils. By contrast, several lysosomal enzymes, lactic dehydrogenase, and plasminogen activator activity, present in cell homogenates, were absent from purified granules.

The granule matrix of guinea pig basophils, unlike the cytoplasmic granules of other granulocytes or mast cells, was little altered by high or low salt concentration but was disrupted into insoluble fragments by 0.01 N HCl and by Triton X-100. Granules were solubilized by papain and by urea-SDS but enzyme activity was destroyed. Triton X-100 incubation with freeze-thawing proved to be the optimal method for extracting esterase activities. Esterase activities were not released from basophils under conditions of anaphylactic degranulation that liberated the great majority of basophil granule histamine.
Effects of α-Fetoprotein on Murine Immune Responses
Authors: Sell S. , Sheppard H. W. , Poler Mark
Published: December 2022

In vitro and in vivo experiments failed to identify a consistent “immunosuppressive” or “immunoregulatory” role for α-fetoprotein (AFP) in the rat. Both normal and AFP-rich sera inhibited Con A, PHA, and MLR proliferation responses of lymphocyte cultures in vitro equally well, in spite of up to 100,000-fold differences in AFP concentration. AFP-rich sera also had no effect on the antibody response of the rat in vivo. Purified AFP from amniotic fluid inhibited the PHA response at concentrations of 100 µg/ml. Purified albumin was also inhibitory, but at 1 log higher concentration. Purified AFP from tumor sera was not inhibitory at 100 µg/ml for Con A, PHA, or MLR reactions and only inhibited PHA stimulation at medium concentrations greater than 1000 µg/ml. The role of AFP as an immunosuppressive agent, particularly in reference to survival of mammalian fetuses, should be reconsidered.
Cytotoxic Effects of Normal Sera on Lymphoid Cells
Authors: Budzko Delia B. , Kierszenbaum Felipe , Waksman Byron H.
Published: December 2022

In the present study, we have examined the identity of the mouse lymphoid cells subject to nonspecific killing by normal guinea pig serum (GPS). In parallel cytotoxicity assays in which the number of cells was varied over a 12-fold range, with all other conditions kept the same, the proportions of cells killed were virtually constant, indicating that a specific cell subpopulation acted as target. Spleen cells surviving treatment with GPS were found to be incapable of responding to the T cell mitogens phytohemagglutinin-P and concanavalin A but responded well to stimulation with endotoxic lipopolysaccharide (LPS), a B cell mitogen, suggesting either a direct or indirect selective effect of GPS on T cells. Similar results were obtained with C4-deficient GPS. LPS-induced DNA synthesis was identical in spleen cells treated with either GPS or anti-Thy-1.2 antibody plus a noncytotoxic source of complement. Spleen cells surviving treatment with anti-Thy-1.2 antibody plus complement were found to be insensitive to GPS cytotoxicity. Conversely, prior incubation of the spleen cells with GPS resulted in the killing of all cells subject to lysis by anti-Thy-1.2 and complement. Spleen cells from nu/nu mice and bone marrow cells from normal mice showed minimal sensitivity to GPS cytotoxicity. A close similarity was observed between the percentages of cells killed by GPS in various lymphoid cell populations and the known proportions of T cells in these populations. In cytotoxicity assays in which normal and T cell-enriched (by passage through nylon wool columns) spleen cell populations were tested with both GPS and anti-Thy-1.2 plus complement, lysis by the two techniques was always comparable. The present results suggest strongly that mouse T cells are selectively killed by GPS and that this cytotoxicity can be used to monitor T cell proportions in mouse lymphoid cell populations with the same efficiency as with the use of anti-Thy-1.2 antibody plus complement.
Suppression of the Human Autologous Mixed Lymphocyte Reaction by Physiologic Concentrations of Hydrocortisone
Authors: Ilfeld David N. , Krakauer Randall S. , Blaese R. Michael
Published: December 2022

Normal human T lymphocytes respond with vigorous proliferation when exposed in vitro to autologous irradiated lymphocytes enriched in B cells. In this report we examined the effect of various humoral factors that might play a role in the regulation of this autologous MLR. Neither fresh autologous plasma, thymic factors, nor hepatoma-derived α-fetoprotein had any significant suppressive effect on the autologous (or allogeneic) MLR. In contrast, over 98% suppression of the autologous MLR occurred with addition of 4 × 10-7 M hydrocortisone succinate. Interestingly, the presence of 2 to 8 × 10-8 M hydrocortisone during the autologous MLC suppressed 62 to 84% of the proliferation, whereas it did not suppress proliferation in the allogeneic MLC. Kinetic studies demonstrated that the predominant suppression of proliferation in the autologous MLC by hydrocortisone occurred during the first few days of culture and that by 100 hr of culture (which is before most of the DNA synthesis occurs) the addition or removal of hydrocortisone had no further effect on the reaction. The presence of hydrocortisone during just the first 6 hr of the autologous MLC also had no suppressive effect on proliferation. Thus, hydrocortisone alters critical functions at a cellular level without causing destruction or other irreversible effects on potentially autoreactive lymphocytes. Furthermore, marked suppression of the autologous MLR without any suppression of the allogeneic MLR was observed with concentrations of free cortisol equivalent to those found in human plasma, suggesting that hydrocortisone may have a physiologic role in the regulation of such T cell autoreactivity in vivo.
Studies of the Mechanism by Which B Cells Permit Mixed Lymphocyte Reactions between Immunocompetent Thymus Cells
Authors: Dyminski J. W. , Smith R. T.
Published: December 2022

The mechanisms by which a critical ratio of peripheral lymphoid helper cells permit a proliferative MLR response between congenic immunocompetent thymus cell subpopulations was investigated. The helper cell was found to be contained in a subpopulation of B cells on the basis of sensitivity to anti-Ig and complement, resistance to anti-Thy-1.2, adherence to nylon columns, and ability to form EA and EAC rosettes with sheep erythrocytes. The maximal helper cell effect was obtained in those combinations in which helper was syngeneic for responder. Helper cells of F1 hybrid origin or syngeneic to stimulator cells were less effective. Kinetic studies indicated that the first of the sequence of events is helper cell-stimulator cell interaction, followed by triggering of responder proliferation generating a subset of cytotoxic cells. Helper activity was trypsin sensitive, and present in solubilized preparation of peripheral cells. Helper activity of intact cells was nearly matched quantitatively by critical ratios of polyclonal IgG, IgG subclasses, and IgG fragment preparations, but little activity was supplied by IgA and IgM preparations. Kinetics studies of IgG helper activity indicate dose-response effects and sequence of interaction similar to those found for whole B cells. A hypothesis is offered to explain how the B-helper or its product may permit an otherwise nonreactive MLR between immunocompetent thymus cells.
A Carbohydrate-Containing Antigen from Trypanosoma cruzi and Its Detection in the Circulation of Infected Mice
Authors: Gottlieb Michael
Published: December 2022

A polysaccharide-containing fraction has been prepared from the water-soluble material of a phenol-water extract of epimastigote forms of Trypanosoma cruzi. Sera from rabbits immunized with this material have been used to detect trypanosome-derived polysaccharide in plasma of mice acutely infected with this parasite. The polysaccharide nature of the circulating antigen is suggested by its heat stability, resistance to proteinase and nucleases, and destruction by sodium metaperiodate, as well as by its periodic acid-Schiff staining and precipitation with concanavalin A. The antigen appears to be present in different strains of the parasite and in different stages in the life cycle of the parasite.
Complement-Mediated Dissociation of Antibody from Immobilized Antigen
Authors: Miller Gary W.
Published: December 2022

Previous studies that showed that complement (C) can convert insoluble Ag-Ab precipitates into soluble immune complexes suggested that activated C components might be able to dissociate Ab from Ag. The present study directly examined this possibility by measuring the dissociation of radiolabeled Ab from immobilized Ag in the presence or absence of active complement. Small polystyrene test tubes were coated with BSA (Ag), and the coating was cross-linked with glutaraldehyde to ensure its stability. Then, 125I-labeled 7S fraction of mouse anti-BSA was allowed to bind to the BSA-coated tubes; controls showed that the binding was specific. The release of labeled Ab from the tubes was measured in the presence of fresh serum or decomplemented serum at 37°C. Active complement markedly increased both the rate of release and the final amount of Ab released. Once begun, the release of Ab could continue for some time in the absence of fluid-phase complement. Controls included duplicate sets of tubes coated with radiolabeled BSA in which unlabeled Ab was bound. These controls showed that, although minute amounts of BSA were unavoidably lost from the tubes, this loss could not account for the observed release of Ab. The ability of the released Ab to rebind to fresh BSA-coated tubes was also examined; it was found that C-released Ab could rebind nearly as efficiently as spontaneously released Ab. The results are completely compatible with the “intercalation” hypothesis developed to explain the C-mediated solubilization of immune aggregates.
Macrophages Suppress CTL Generation in Rat Mixed Leukocyte Cultures
Authors: Weiss Arthur , Fitch Frank W.
Published: December 2022

The generation of CTL in rat MLC was actively suppressed by a cell population present in spleen cell preparations from normal rats. These suppressor cells were characterized by using a variety of cell fractionation techniques. Suppressor cells were removed by passage of spleen cells through nylon wool columns or by treatment with carbonyl iron. Suppressive activity was present in the mononuclear cell fraction of spleen cells obtained by Ficoll-Hypaque density gradient centrifugation. After velocity sedimentation at unit gravity, enrichment of suppressive activity was demonstrated in the fractions containing large cells as compared to the fractions containing small cells. Populations rich in macrophages were shown to have similar suppressive activity upon CTL induction in MLC. These studies suggest that macrophages present in normal rat spleen cell preparations account for the difficulty in generating CTL in MLC prepared with rat cells.
Transient Suppression of the Humoral Immune Response Mediated by a Factor Derived from Specifically Activated, Doubly Primed Lymphoid Cells
Authors: Kempf Kenneth E. , Rubin Arnold S.
Published: December 2022

In cultures of sheep erythrocyte- (SRBC) stimulated spleen cells from mice immunized with tetanus toxoid (TT) and horse erythrocytes (HRBC) 30 to 90 days earlier, the addition of both HRBC (day 0) and TT (day 2) resulted in significant suppression of the anti-SRBC plaque-forming cell (PFC) response compared to the response of similar cultures maintained without the priming antigens. The observed inhibition was due to the presence of a soluble factor that was released into the supernatant fluid of the specifically stimulated, primed population of lymphoid cells between 72 and 120 hr after culture initiation. The active mediator, a macromolecule of ∼∼24,000 daltons as determined by gel filtration over Sephadex G-150 and Ultrogel AcA 44, was suppressive when added within 24 hr, but not 48 hr, of assay for PFC against the reference SRBC antigen. The transiently acting soluble suppressor (TASS) was not overtly cytotoxic since total cell recovery and viability were unaffected in its presence. The results presented here are discussed in relation to a possible mechanism of action in which the negative regulation of immunoglobulin production is favored once a minimum level of immune reactivity is reached.
Enhancement of the PGE1 Response of Macrophages by Concanavalin A and Colchicine
Authors: Gemsa Diethard , Steggemann Ludwig , Till Gerd , Resch Klaus
Published: December 2022

The effects of concanavalin A (Con A) and colchicine on the prostaglandin E1 (PGE1)-mediated cyclic AMP generation in rat peritoneal macrophages have been studied. Although Con A and colchicine by themselves did not affect cyclic AMP levels, they greatly enhanced cyclic AMP production induced by PGE1. There was not only augmentation of cyclic AMP levels at maximally active concentrations of PGE1, but also an increased sensitivity to low (inactive) concentrations of PGE1. Except for lentil lectin, none of the other lectins affected PGE1 sensitivity whereas lumicolchicine was as effective as colchicine. In addition, both Con A and colchicine raised the sensitivity to isoproterenol and choleraenterotoxin. Although details of the mechanisms by which Con A or colchicine influenced the membrane-bound adenyl cyclase and PGE1 receptors remain unclear, these observations suggest that certain alterations of the cell membrane may render macrophages more susceptible to the regulating effects of prostaglandins.