bluuebunny/crossref_metadata_2025 huggingface dataset:
Human Macromolecular Insoluble Cold Globulin (MICG)
Authors: Hauptman Stephen P. , Kansu Emin , Sobczak Gloria , Serno Maryann
Published: December 2022

Thymus cell-derived macromolecular insoluble cold globulin (T-MICG) is a 225,000-dalton protein, selectively synthesized in human T cells. Null cell-derived macromolecular insoluble cold globulin (N-MICG) is a 185,000-dalton protein, synthesized in null cells, and antigenically distinct from T-MICG. Evidence to support these conclusions was provided by using isolated cell preparations that were radiolabeled, lysed in desoxycholate, and precipitated with monospecific antiserum to each component. These studies demonstrated that antiserum to T-MICG precipitated a 225,000 dalton protein from PBL and T cells, but not from B or null cells. Antiserum to N-MICG reacted with a 185,000 dalton protein present in PBL and null cells, but not with lysates from either T or B cells.

The plasma membrane distribution of these proteins was shown by absorption of antiserum to T + N-MICG with either isolated T or null cells. Antibody-induced cytotoxicity and immunofluorescence confirmed the cell surface location of T and N-MICG.

Divergent biologic effects of these antisera were also noted. Antiserum to T-MICG inhibited T cell rosette formation and the one-way mixed lymphocyte reaction, although anti-N-MICG antiserum had no such effect. The potential importance of these proteins is discussed.
Human Leukocyte Inhibitory Factor (LIF): Two Distinct Molecular Species
Authors: Brown Alan P. , Rocklin Ross E.
Published: December 2022

Human leukocyte inhibitory factor or LIF was generated in vitro by stimulating blood lymphocytes with concanavalin A (Con A). The control and Con A active supernatants were partially purified by gel filtration on Sephadex G-100. The fraction containing LIF (68,000 daltons) activity was then subjected to isoelectric focusing (pH 3 to 10 ampholines) in a sucrose gradient. Two LIF activities were reproducibly recovered by this procedure. One molecular form was found to have an isoelectric point of approximately pH 5.0 and the other approximately pH 8.5. Both molecular species were rechromatographed on Sephadex G-75 and found to have the same apparent m.w. (68 to 75,000). Furthermore, the biologic activity of both factors was destroyed after treatment with diisopropylphosphofluoridate, suggesting that they may be esterases.
Immunosuppressive Activity of Canine Pulmonary Surface Active Material
Authors: Ansfield Michael J. , Kaltreider H. Benfer , Benson Bradley J. , Caldwell Joseph L.
Published: December 2022

The effects of canine surface active material (SAM), a lipoprotein unique to the lung, on the proliferative response of lymphocytes to immune stimulation were investigated. Peripheral blood lymphocytes were cultured in the presence of either endobronchial lavage fluid or purified SAM extracted from such lavage fluids. Cultures were stimulated with mitogens (Con-A, or PWM) or allogeneic cells (MLC), and the proliferative responses were measured by the uptake of tritiated thymidine.

Culture of lymphocytes in autologous endobronchial lavage fluid resulted in a suppressed response to all three stimuli: MLC (33% control response), PWM (54% control), Con-A (75% control). Culture of lymphocytes with purified SAM produced a dose-dependent inhibition of the responses to these stimuli (MLC > PWM > Con-A). Fifty percent inhibition of lymphocyte responses to MLC, PWM, or Con-A was produced by 4.2, 7.5, or 12.5 µg protein/ml of purified SAM, respectively. Supraoptimal doses of mitogens failed to alter the observed inhibitory activities. SAM was not cytotoxic for lymphocytes in these experiments. SAM was most inhibitory when added to lymphocytes at the time of mitogen stimulation; progressively less inhibition was produced by SAM added at increasing intervals of time after stimulation. Lymphocytes preincubated with SAM for 24 or 48 hr, washed, and then cultured in fresh medium displayed persistently suppressed responses.

These results demonstrate that canine SAM is a potent suppressor of lymphocyte proliferation in vitro. In vivo, SAM is found only in the lung and, during fetal development, is secreted by the lung into amniotic fluid. The presence of SAM in these locations may have important implications for our understanding of pulmonary and maternal-fetal immunology.
Susceptibility of Friend Virus Antigen-Modulated Erythroleukemic Cells to Lysis by T Lymphocytes from Mice with Dormant Friend Virus Infections
Authors: Genovesi Eugene V. , Marx Preston A. , Wheelock E. Frederick
Published: December 2022

Spleen cells from DBA/2 mice with dormant Friend leukemia virus (FLV) infections or from mice immunized with x-irradiated FLC-745 erythroleukemic cells are not cytolytic for FLC-745 cells when tested directly, but acquire cytolytic activity in vitro by cultivation with x-irradiated FLC-745 cells. Spleen cells cultured from normal mice do not acquire such cytolytic activity. Cytolytic activity resides in the T lymphocyte population. Alloantiserum, but not antisera against FLV-virion polypeptides, inhibited the lysis of FLC-745 cells by cytolytic T lymphocytes. To understand further the role of cellular and humoral immune anti-FLV responses in mice with dormant FLV-infections, in vitro experiments were conducted that mimicked the in vivo FLV-specific immune environment of these mice. We found that FLV-immune serum from mice with dormant FLV-infections modulated FLV-antigen expression on the surfaces of FLC-745 cells without affecting their susceptibility to lysis by cytolytic T lymphocytes. These results suggest that cytolytic T lymphocytes restrain the outgrowth of FLV-antigen-modulated erythroleukemic cells in mice with dormant FLV infections and that the targets for cell-mediated lysis may be H-2d-associated antigens.
A Reexamination of Humoral Tolerance in Chickens Congenitally Infected with an Avian Leukosis Virus
Authors: Qualtiere Louis F. , Meyers Paul
Published: December 2022

In a reexamination of congenital infection of chickens with ALV, one of the classical models of immunologic tolerance, we were unable to demonstrate that there was any evidence suggesting humoral immune reactivity to the infecting virus. In ALV-F42 congenitally infected birds that have a persistent viremia and no neutralizing antibody detectable by conventional means, we could find no evidence of host IgG deposits in the kidneys, nor any suggestion of renal pathology or damage. In addition, attempts to precipitate any putative circulating infections virus-antibody complexes by treatment with a highly specific anti-globulin were negative nor were there titer differences in the viremic plasmas of bursectomized birds compared to nonbursectomized birds, indicating that the chickens were not making either a masked neutralizing or a nonneutralizing antibody response to the infecting virus. The evidence presented indicates that congenital infection of chickens with ALV does, at least at the humoral level, result in a state of immunologic tolerance to the infecting agent.
Genetic Control of the Immune Response to Staphylococcal Nuclease
Authors: Pisetsky David S. , Riordan Stephen E. , Sachs David H.
Published: December 2022

The genetic linkage relationship of two antinuclease idiotypes produced by the BALB/c strain was investigated in the backcross (BALB/c × CB.20) × CB.20. These two idiotypes were detected by Lewis rat anti-idiotypic antisera prepared against affinity-purified A/J and SJL antinuclease antibodies, termed the A/J and SJL idiotypes, respectively. Both idiotypes were found to be linked to the IgCHa immunoglobulin heavy chain allotype locus. There was, however, a high frequency of recombination observed between both markers and the IgCHa locus, with eight of 83 backcross animals recombinant for the A/J idiotype and five of 83 recombinant for the SJL idiotype. All such recombinant animals were IgCHb/b homozygotes that had gained one or both idiotypes. These results are consistent with a genetic map of VH region genes in the BALB/c strain in which genes determining the SJL idiotype are closer to the IgCHa allotype locus than are genes determining the A/J idiotype. This high frequency of recombination may indicate that the chromosome segment containing VH region genes is very large or that it has structural features that promote recombination.
Cell Types Required for H-2-Restricted Cytotoxic Responses Generated by Trinitrobenzene Sulfonate-Modified Syngeneic Cells or Trinitrophenyl-Conjugated Proteins
Authors: Pettinelli Carla B. , Schmitt-Verhulst Anne-Marie , Shearer Gene M.
Published: December 2022

Murine spleen cells were fractionated over nylon wool or Sephadex G-10 columns, and the cell types involved in the generation of trinitrophenyl (TNP)-specific, H-2 restricted (TNP-self) cytotoxic effector cells were studied from cultures stimulated with trinitrobenzene sulfonate (TNBS)-modified syngeneic cells, TNP-conjugated soluble proteins such as bovine γ-globulin (TNP-BGG), or bovine serum albumin (TNP-BSA). Unfractionated or nylon nonadherent responding cells generated such effectors, irrespective of whether the cultures were stimulated with TNBS-modified cells or TNP-conjugated proteins. TNP-modified T lymphocytes, B lymphocytes, and phagocyte-enriched spleen cells were all capable of stimulating TNP-self effectors. TNP-self as well as allogeneic cytotoxic responses were dependent on the presence of a radioresistant non-T cell that was removed by Sephadex G-10 fractionation and was replaced by irradiated, Thy 1.2-negative, glass adherent spleen cells, enriched in phagocytic cells. Results obtained by using glass adherent cells that were allogeneic or semi-syngeneic to the responding cells indicated that H-2 homology was not required for efficient glass adherent cell function, and that the H-2 restriction of TNP-self effectors is not determined by these glass adherent cells.
Failure of NZB Spleen to Respond to Prethymic Bone Marrow Suppressor Cells
Authors: Dauphinée Michael J. , Talal Norman
Published: December 2022

Mouse bone marrow contains θ-negative lymphocytes that can suppress an in vitro plaque response by spleen cells primed in vivo with burro red blood cells (BRBC). These bone marrow cells are radiosensitive and can be induced with thymosin fraction 5 or α1 thymic peptides to express the θ antigen. Enrichment for these suppressor pre-T lymphocytes can be achieved by a one-step density centrifugation, macrophage depletion, or a combination of both procedures.

NZB mice, which spontaneously develop an autoimmune disorder, have a suppressor abnormality revealed by this assay system. Upon analysis, they have normal BM pre-T suppressor cells but their spleen cells are refractory to the BM suppressor signal. NZB BM suppressor cells inhibit the response by DBA/2 spleen cells, but DBA/2 BM suppressor cells do not inhibit NZB spleen. This resistance to suppression is a property of the B cell fraction recovered from NZB spleen.
Study on the Turnover of the Receptor for the Third Component of Complement on Human Lymphoid Cells
Authors: Rosso di San Secondo V. E. M. , Meroni P. L. , Fortis C. , Tedesco F.
Published: December 2022

The in vitro turnover of the receptor for the third component of complement (C3) was studied in normal peripheral blood lymphocytes (PBL) and in lymphoblastoid cells from established cell cultures of both “normal” and “malignant” origin. The turnover was evaluated by studying i) the disappearance rate of the C3-receptor in cells in which the protein synthesis was blocked by cycloheximide and puromycin, ii) the reexpression rate of the C3-receptor after treatment of the cells with either rabbit antiserum against B lymphocytes or mouse C activated through the alternative pathway by inulin. The results show that the C3-receptor of all the lymphoid cells has roughly a half-life of about 3 to 4 hr. However, the cultured lymphoblastoid cells were less sensitive than normal PBL to inhibition by cycloheximide and showed a faster reexpression rate of the C3-receptor. A spontaneous release of the receptor was found to occur, since a receptor-like activity was detected in the spent culture medium of long-term cultured lymphoid cells.
Immunoregulatory Circuits among T Cell Sets: Effect of Mode of Immunization on Determining Which Lyl T Cell Sets Will be Activated
Authors: Eardley D. D. , Kemp J. , Shen F. W. , Cantor H. , Gershon R. K.
Published: December 2022

Lyl T cells are able to induce B cells to make antibody and also to induce a resting Ly123 T cell set to exert potent feedback suppression. Which of these two pathways Lyl T cells take can be influenced by the mode of immunization. In particular, in vitro immunized Lyl T cells are more likely to induce suppression than are Lyl T cells immunized in vivo even when both populations deliver the same amount of help to purified B cells. The feedback suppression induced by the Lyl T cells immunized in vitro can be distinguished from suppression mediated by Ly2+ T cells; in the former case suppression is preceded by a precocious antibody response, whereas suppression mediated by Ly2+ T cells is apparent throughout the entire period of observation.
Naturally Occurring Circulating Immune Complexes: Normal Human Serum Contains Idiotype-Anti-Idiotype Complexes Dissociable by Certain IgG Antiglobulins
Authors: Morgan Alton C. , Rossen Roger D. , Twomey Jeremiah J.
Published: December 2022

Sera from 19 of 23 healthy subjects contained a 24 to 26S complex (NTC). When passed over heat-aggregated IgG (HAGG) Sepharose, the NTC complex dissociated into a 7S IgG that was cytotoxic for malignant melanoma cells, and into an IgG antiglobulin (NTC antiglobulin) that only reacted with autologous cytotoxin. Prior incubation with soluble extracts of melanoma cells, but not with extracts of colon cancer or sarcoma, blocked recombination of isolated cytotoxin with NTC antiglobulin. Thus, NTC antiglobulin had attributes of an anti-idiotype. Cohn Fr II contained NTC complexes from many donors. A second IgG antiglobulin (ACF) was found in sera from patients with advanced cancer or rheumatoid arthritis. When added to normal donor sera, ACF caused NTC antiglobulin and 7S cytotoxin to dissociate. ACF combined with Fab′ of the allogeneic NTC cytotoxin to create a new complex that facilitated complement-dependent cytolysis of malignant melanoma cells. Sera with ACF activity also contained NTC. However, the cytotoxin in these sera was not detectable until the endogenous ACF and NTC antiglobulin had been removed by adsorption to HAGG Sepharose.

In ACF-containing sera, ACF fixed to NTC antiglobulin but would not react with the cytotoxin. In contrast, ACF reacted with the cytotoxin but not NTC antiglobulin in allogeneic sera. Thus, some natural antibodies circulate with their antigen-combining sites blocked by endogenous antiglobulins. Other antiglobulins, such as ACF, can facilitate detection of these blocked antibodies if they react with the components of the complex in a way that facilitates their dissociation.
Limiting Dilution Analysis of Alloantigen-Reactive T Lymphocytes
Authors: Ryser Jean-Etienne , MacDonald H. Robson
Published: December 2022

A sensitive mixed leukocyte microculture (micro MLC) system has been developed in which both proliferative and cytolytic responses can be detected from small numbers of responding alloantigen-reactive T lymphocytes. Cytolysis and proliferation in micro MLC were assessed by conventional 51Cr release and 3H-thymidine uptake assays, respectively. Optimal sensitivity of the system was dependent on several parameters, the most important of which was the presence of supernatant fluid derived from secondary MLC (2° MLC SN). Under optimal conditions, formal limiting dilution analysis could be achieved since excellent discrimination between positive and negative microcultures was obtained for both the cytolytic and proliferative assays. The system was therefore used to obtain simultaneously minimal estimates of the frequency of cytolytic T lymphocyte precursors (CTL-P) and of the precursors of proliferating T lymphocytes (PTL-P). The results of a series of experiments involving C57BL/6 spleen cells responding against DBA/2 alloantigens indicated that the minimal frequency of CTL-P was approximately 2-fold lower than the frequency of PTL-P in the same responding cell population (average of 1:570 and 1:290, respectively). Since analysis of individual micro MLC further demonstrated that essentially all CTL-P were contained within the PTL-P population, the simplest interpretation of these findings would be that approximately 50% of alloantigen-reactive T cells in this strain combination have the capacity to give rise to specific CTL. Some applications of this model system are briefly discussed.
Moloney Virus-Induced Cell Surface Antigens and Histocompatibility Antigens Are Located on Distinct Molecules
Authors: Fox Robert I. , Weissman Irving L.
Published: December 2022

The Moloney lymphoma, LSTRA, is widely used as a target cell for cell-mediated lysis studies. This report further characterizes the antigenic structures that may be important in the cytotoxic T lymphocyte-target cell recognition. Anti-H-2 sera have been absorbed to remove contaminating antiviral activity. The results presented demonstrate that: a) no linkage between H-2 and viral antigens was detected with 1-dimensional (SDS-PAGE) or 2-dimensional (IEF followed by SDS-PAGE) gel electrophoresis; b) no molecule showing the antigenic properties of both viral and histocompatibility antigens was detected; c) H2-Dd and H2-Kd molecules on the LSTRA lymphoma appeared antigenically similar to the parental BALB/c cells based on inhibition of cytotoxicity experiments; d) with cell extracts made in the presence of iodoacetamide, no disulfide linkages could be demonstrated between viral and histocompatibility antigens; e) the Moloney viral antigen gp70 was found in large amounts on the cell surface of LSTRA cells; f) the p30 precursor (gag) was not detectable on the cell surface, although it was abundant in the cell cytoplasm.

Thus, these studies indicate that the LSTRA target cell appears to have H-2 and viral antigens present on separate molecules, which are dissociated during NP40 detergent extraction. No evidence was found of an “altered self” type molecule, in which either H-2 or viral antigens had been chemically altered as a consequence of viral transformation.
Differences in HLA Antigen Recognition by Human Influenza Virus-Immune Cytotoxic T Cells
Authors: Biddison William E. , Shaw Stephen
Published: December 2022

The specificity of in vitro induced human influenza-immune cytotoxic effector cells was analyzed with respect to recognition of HLA-A and -B-linked gene products. The influenza-immune cytotoxic activity observed on panels of virus-infected targets demonstrated that virus-immune effectors preferentially lyse targets with which they share HLA-A or -B specificities. Virus-immune effectors from certain donors recognized virus in conjunction with some, but not all, of their self HLA-A and -B antigens. Among donors who share a given HLA antigen (such as A2 or B7), there are differences in the ability of their virus-immune T cells to recognize the shared antigen. Virus-infected target cells from HLA-A2 or -B7 “nonresponder” donors could be lysed by virus-immune T cells obtained from other donors who shared only the HLA-A2 or -B7 antigen with these target cells.

These observations suggest that the absence of cytotoxic T cell responses by some donors to influenza virus in conjunction with HLA-A2 or -B7 is not due to control by the structural genes that code for these HLA antigens, but rather may result from control by regulatory genes that act at the level of the responder and/or stimulator cell. The results are discussed in the context of Ir gene regulation of human T cell responses.
Mechanisms of Mouse Mast Cell Activation and Inactivation for IgE-Mediated Histamine Release
Authors: Siraganian Reuben P. , Hazard Kimberley A.
Published: December 2022

The IgE-mediated histamine release from mouse mast cells requires Ca++, is optimal at 37°C, and is enhanced by phosphatidylserine. The rate of release is relatively slow. The mast cells can be activated to release histamine by either anti-IgE or anti-Fab antibodies and, in the case of cells from sensitized mice, by the immunizing antigen. The incubation of mast cells with antigen in the absence of Ca++ or phosphatidylserine fails to release histamine. Such cells are desensitized to the further addition under optimal conditions of the same antigen. Desensitization is antigen specific, requires optimal levels of antigen, and occurs at both 30° and 37°C. In contrast, anti-IgE desensitizes all IgE-mediated histamine release reactions.
The Structural Proteins of Murine Leukemia Virus and the Pathogenesis of Necrotizing Arteritis and Glomerulonephritis in SL/Ni Mice
Authors: Yoshiki Takashi , Hayasaka Takashi , Fukatsu Ryo , Shirai Toshikazu , Itoh Tetsuo , Ikeda Hisami , Katagiri Makoto
Published: December 2022

The analysis by immunofluorescent and electronmicroscopic studies of murine leukemia virus (MuLV), which could be of significance in the pathogenesis of necrotizng arteritis and glomerulonephritis in SL/Ni mice, yielded the following conclusions. The structural proteins, gp70, p30, and p15(E) of MuLV were granularly deposited along with bound immunoglobulins and C3, apparently as immune complexes, in the vessel walls of necrotizing arteritis. Just before or at the appearance of vascular lesions, C-type particles were found in greatest abundance around muscular cells and could be seen budding from the membrane of these cells. There was significant correlation between the presence of aggregated degenerating C-type particles and the presence of dense deposits in those vascular walls. In contrast, gp70 was the major viral protein deposited along with host immunoglobulins and C3 in the diseased kidney of mice. Electron dense deposits were demonstrated in the affected glomeruli.

In attempts to identify directly the molecule(s) bearing the antigenic determinants for Gross natural antibody (GNA), 125I-labeled E♂G2 MuLV proteins were subjected to immunoprecipitation with GNA-positive mouse sera. SDS-PAGE analysis of the precipitates revealed that the major molecule bearing antigenic determinants or GNA is p(85), the precursor polyprotein of the MuLV core structural proteins, p30, p15, and p10.
Shared Chemical Properties of Different Murine Thymus-Leukemia Antigens
Authors: Pischel Ken D. , Little J. Russell
Published: December 2022

Immunochemical studies of murine thymus-leukemia antigens (TLA) have confirmed that the subunit structure consists of a 45,000-dalton heavy chain and a β2 microglobulin (β2m) light chain. Similar structural features are exhibited by the TLA from thymocytes of Tlaa, Tlac, Tlad, and a leukemia cell derived from C57BL/6, a Tlab strain. In addition to the similar subunit structure from the four haplotypes, each TLA shows a similar pattern of trypsin proteolysis. This procedure yields a major heavy chain cleavage product of approximately 37,000 daltons that remains associated with β2m and retains most or all of the antigenic determinants of the intact TLA. Evidence is presented that TLA do not exhibit Fc receptor properties, nor do they adsorb to murine leukemia virus antigens under the conditions of isolation for analysis on polyacrylamide gel electrophoresis (PAGE) in sodium dodecyl sulfate (SDS). Taken together these findings strongly support the hypothesis that TLA comprise a family of chemically similar antigens belonging to a structurally and genetically related group that includes H-2D, H-2K, and Qa-2,3.
Antibody Penetration Into Living Cells
Authors: Alarcon-Segovia Donato , Ruiz-Arguelles Alejandro , Llorente Luis
Published: December 2022

We have previously shown that an anti-ribonucleoprotein (RNP) IgG can penetrate into live human mononuclear cells (MNC) having receptors for the Fc portion of IgG. Because T cells with such receptors (Tγ cells) seem to behave as suppressor cells in immune regulation and because this suppressor function is diminished in diseases where antinuclear antibodies appear, we considered the possibility that antinuclear IgG antibody could penetrate Tγ cells and affect them.

Herein we show that fluorescein-labeled anti-RNP IgG can penetrate into Tγ cells, enriched by either mitogenic stimulation or separation with a subpopulation of T cells with low affinity for sheep erythrocytes. Incubation of MNC with anti-RNP IgG before carrying out the separation procedures resulted in apparent loss of Tγ cells at the end of separation. To confirm that deletion had actually occurred, we performed a cytotoxicity assay using 51Cr-labeled T cells. Anti-RNP IgG had a significantly higher cytotoxic effect than normal IgG on T cells, particularly on those with low affinity for sheep erythrocytes that include most Tγ cells.

Suppressor cell function studied in a system where it was expanded, by either 7-day culture or incubation with concanavalin A, and detected in a reverse plaque-forming cell assay with rabbit anti-human immunoglobulin-developing antibody was found to be abrogated by the addition of anti-RNP IgG to the suppressor function-expanding cultures.

Controls in Ig-free medium, or medium supplemented with normal human IgG, aggregated normal human IgG, BSA-anti-BSA immune complexes, or F(ab′)2 fragments of the anti-RNP IgG, did not abrogate suppressor cell function. This indicates that the abrogation of suppressor cell function by anti-RNP IgG is due to its penetration into Tγ cells. Suppressor cell loss and/or dysfunction caused by penetration of antinuclear antibodies into Tγ cells may lead to the self-perpetuation of autoimmune disease.
Carboxymethyl Cellulose, a Nonimmunogenic Hapten Carrier with Tolerogenic Properties
Authors: Diner Uriel E. , Kunimoto Dennis , Diener Erwin
Published: December 2022

This communication reports on the tolerogenic properties of carboxymethyl cellulose (CMC) as a nonimmunogenic carrier for 2.4 dinitrophenyl (DNP) and the benzylpenicilloyl determinant (BPO). Either normal or primed mice, given an optimal dose of 250 µg per animal of DNP CMC, when challenged with an immunogenic form of the hapten as early as 30 min or as late as 21 days thereafter were completely and specifically unresponsive to it. Experimental evidence suggests that this unresponsiveness is not due to suppressor cells. Furthermore, DNP CMC induces tolerance in vivo but fails to do so in vitro under conditions at which other tolerogenic carbohydrate hapten conjugates such as DNP-dextran do. This together with comparative studies of tolerance induction kinetics by DNP CMC and DNP-dextran in vivo led us to conclude that molecular properties other than the epitope density must be attributed to CMC's tolerogenic potential. CMC may also be used as a tolerogenic carrier for BPO with respect to IgE antibody production. Thus, normal or primed mice injected with the BPO CMC conjugate were found specifically unresponsive to a challenge with an immunogenic form of penicillin.
Adaptive Differentiation of Murine Lymphocytes
Authors: Katz David H.
Published: December 2022

Responses to the synthetic terpolymer L-glutamic acid, L-lysine, L-phenylalanine (GLΦ) and hapten derivatives thereof are controlled by two complementing H-2-linked Ir genes in the mouse. F1 hybrids derived from two different nonresponder strains (one of which possesses the α and the other β Ir-GLΦ gene) are phenotypic responders to GLΦ and 2,4-dinitrophenyl (DNP)-GLΦ. Moreover, spleen cells from DNP-GLΦ-primed F1 mice can adoptively transfer secondary anti-DNP antibody responses to irradiated F1 recipients that have been challenged with DNP-GLΦ. When, however, GLΦ-primed F1 helper T cells are transferred together with the DNP-specific F1 B cells that had been primed in separate mice altogether by DNP coupled to an unrelated protein carrier, such mixtures failed to develop adequate adoptive secondary anti-DNP responses to DNP-GLΦ. This contrasted with the ability of the same GLΦ-primed F1 T cells to provide helper activity for DNP-primed B cells from responder recombinant B10.A (5R) mice. More important, the apparent defect of GLΦ-primed F1 T cells in providing help for DNP-primed F1 B cells (primed to a DNP-protein conjugate) could be readily overcome by using DNP-primed B cells from donor F1 mice primed with DNP-GLΦ. As discussed herein, these results suggest that interacting T and B lymphocytes pair off into partner cell sets, any pair of which interact optimally when a “best fit” reciprocal self-recognition occurs between them.
Mechanism of Immunosuppression of Progesterone on Maternal Lymphocyte Activation during Pregnancy
Authors: Clemens Louis E. , Siiteri Pentti K. , Stites Daniel P.
Published: December 2022

Lymphocyte reactivity was measured in human mixed lymphocyte (MLC) and mitogen-stimulated cultures in the presence of various steroids. When present during the entire culture period at concentrations between 1 and 20 µg/ml progesterone, estradiol, and testosterone blocked 3H-thymidine (3H-TdR) incorporation. Estriol and 16 α-hydroxy progesterone were without suppressive effects. The kinetics of steroid mediated suppression of thymidine incorporation and DNA synthesis was investigated at various times during lymphocyte activation. When progesterone was present only at the beginning of culture, subsequent DNA content, but not thymidine incorporation was suppressed. However, if added late during the activation process, progesterone reduced thymidine incorporation independent of cellular DNA content. If present during the entire culture period both were suppressed. Cortisol, but not progesterone irreversibly inhibited lymphocyte thymidine incorporation if added during the first 12 hr of culture. A reversible blockade of lymphocyte thymidine uptake could be observed with progesterone even during the final hours of mitogen or MLC cultures. Thus, effects of steroid hormones on cellular DNA synthesis and thymidine incorporation appear to be independent processes. Cell fractionation experiments demonstrated that progesterone blocked the uptake of 3H-TdR into the lymphocyte nucleoside pool thereby reducing its availability for incorporation into DNA. This study is consistent with the hypothesis that lymphocyte proliferation at the maternal fetal-interface during pregnancy may be under specific regulation by progesterone. The relatively high local concentrations of the hormone produced by trophoblast are immunosuppressive whereas the substantially lower peripheral steroid levels are entirely inadequate to suppress systemic immunity. We suggest that at least one important role of progesterone in maintaining pregnancy is its regulation of cellular immunity in the maternal-fetal bed.
Hyperthermia and Human Leukocyte Function
Authors: Roberts Norbert J. , Sandberg Kathryn
Published: December 2022

Fever has been shown by others to have a beneficial effect on the survival of animals challenged with bacteria. The febrile animals exhibited increased leukocyte accumulation at the site of bacterial inoculation. In order to investigate further these observations, as well as their possible relevance to human temperature elevation, human leukocytes were assayed in vitro for production of and response to leukocyte migration inhibition factor (LIF) at normal (37°C) and hyperthermic (38.5°C) temperatures. Leukocyte migration was assayed at both 37° and 38.5°C, by using an agarose plate technique, with exposure of the cells to supernatant fluids derived from stimulated and unstimulated leukocytes incubated at 37 or 38.5°C.

Hyperthermia (38.5°C) was consistently associated with greater spontaneous and stimulated LIF production by human mononuclear leukocytes. Hyperthermia was associated with decreased spontaneous leukocyte migration and greater inhibition of leukocyte migration by LIF-rich supernatant fluids. Migration at 38.5°C after exposure to LIF produced at 38.5°C was greatly reduced relative to migration at 37°C in response to LIF produced at 37°C (p <0.005). Results were similar with mitogen- and antigen-stimulated LIF. The results suggest one possible mechanism by which hyperthermia may benefit the
Immunoregulation of Murine Myeloma: Isologous Immunization with M315 Induces Idiotype-Specific T Cells that Suppress IgA Secretion by MOPC-315 Cells in vivo
Authors: Rohrer James W. , Odermatt Bernhard , Lynch Richard G.
Published: December 2022

Previous studies demonstrated that MOPC-315 cells enclosed in peritoneal diffusion chambers (DC) in M315-immunized BALB/c mice did not secrete the 2,4,6-trinitrophenyl (TNP)-binding IgA paraprotein (M315). The secretory blockade was: i) idiotype (Id315)specific, ii) reversed in vitro after mild pronase treatment, and iii) independent of any effect on the cytologic differentiation of MOPC-315 from lymphocytoid to plasmacytoid cells. In addition to secretory blockade, MOPC-315 cells in M315-immunized hosts exhibited a marked reduction in expression of cell surface M315 (sIgA315). Since the alterations in M315 expression occurred across a 0.1-µ Millipore membrane, we proposed that Id315-specific, soluble factors contributed to the effector mechanism.

In the present studies, Id315-specific suppression of M315 secretion was adoptively transferred to normal mice with spleen cells from M315-immunized donors, and the transfer could be abrogated by treatment of the immune spleen cells with heterologous anti-ϑ serum plus complement. Furthermore, in contrast to actively immunized mice, adoptively immunized mice neither developed anti-Id315 antibodies nor suppressed sIgA315 expression.

These findings: i) establish that idiotype-specific suppression of M315 secretion is a process distinct from inhibition of sIgA315 expression, ii) demonstrate that secretory blockade of MOPC-315 cells is dependent on Id315-specific T cells, and iii) suggest that inhibition of sIgA315 expression is dependent on anti-Id315 antibodies.

The results presented here establish that immunoglobulin expression by MOPC-315 cells is responsive to idiotype-specific T cell regulation as well as the antigen-specific T cell regulation we described earlier. These studies provide further insight into the mechanisms of idiotype-specific regulation of myeloma and may contribute to more complete understanding of T cell-mediated regulation of idiotype expression in non-neoplastic B cells.
Genetic Control of in vivo Sensitivity to Lipopolysaccharide: Evidence for Codominant Inheritance
Authors: McGhee Jerry R. , Michalek Suzanne M. , Moore Robert N. , Mergenhagen Stephan E. , Rosenstreich David L.
Published: December 2022

The genetic control of several endotoxin-induced in vivo effects, specifically, immunogenicity, adjuvanticity, interferon production, and lethality, was studied. Lipopolysaccharide (LPS) responder (C3H/HeN or C57BL/6J) mice produced high levels of anti-LPS antibody in response to an optimal dose (10 µg) of a phenol-water extracted LPS from Escherichia coli K235 (Ph-LPS), whereas nonresponder (C3H/HeJ) mice did not demonstrate significant antibody responses. The F1 progeny of responder C3H/HeN × C3H/HeJ parents exhibited low to intermediate responses. C3H/HeN, F1 and C3H/HeJ mice produced equivalent amounts of antibody in response to endotoxin prepared by butanol-water extraction. Adjuvanticity was studied by measuring the immune response to trinitrophenyl group (TNP) coupled to homologous erythrocytes (MRC). Ph-LPS was a good adjuvant for TNP-MRC in C3H/HeN mice, whereas no response was observed with C3H/HeJ mice; the adjuvant response in F1 hybrid mice was intermediate. Similarly, interferon production induced by optimal doses of Ph-LPS followed the same pattern. Finally, F1 hybrid mice were clearly intermediate in sensitivity to LPS-induced lethality. The results of this study demonstrate that the in vivo responses of mice to LPS, like the in vitro sensitivity of lymphoid cells, are under codominant gene control.
In vitro Stimulation of Natural Killer (NK) Cells by Soluble Factor(s) Generated in Antigen-Stimulated Rhesus Monkey Peripheral Blood Lymphocyte Cultures
Authors: Lazda Velta A. , Baram Peter
Published: December 2022

Peripheral blood lymphocytes from rhesus monkeys (Macaca mulatta) sensitized to keyhole limpet hemocyanin (KLH), when stimulated in vitro with KLH, developed natural killer (NK) cell activity that was assayed with Rous Sarcoma virus-transformed marmoset fibroblasts as targets in a 4-hr 51Cr-release assay. The supernatant fluids from 24- to 25-hr KLH-activated cultures were capable of stimulating NK development in nonsensitive lymphocyte cultures. The effector cells were neither macrophages nor B cells (plastic and nylon-wool nonadherent) and did not form E-rosettes with neuraminidase-treated sheep red blood cells. Cultures depleted of EA-rosetting cells, i.e., Fc-receptor-bearing lymphocytes, were incapable of generating NK activity when stimulated in vitro. Kinetic studies showed that peak DNA synthesis, as measured by 3H-T incorporation, preceded maximum cytotoxicity. Elimination of dividing cells by 5-bromo-2′-deoxyuridine (BrdU) and light treatment during the interval from day 1 to day 4 inhibited the development of cytotoxicity on day 7. Cell replication was required for the induction of NK cells with KLH as well as with antigen-activated culture supernatant fluids. When cultures were left unstimulated for 4 days, NK activity could not be developed subsequently either by adding antigen, mitogen (PHA), or supernatant fluids from activated cultures.
Effect of Purified Protein SAA on Immune Response in vitro: Mechanisms of Suppression
Authors: Benson Merrill D. , Aldo-Benson Marlene
Published: December 2022

Studies were done to characterize the immunosuppressive effect of the serum α-globulin SAA. The in vitro antibody response of CBA/J murine spleen cells was studied in the Marbrook culture system. Addition of 10% SAA-containing serum from casein-treated CBA/J mice suppressed the primary antibody response to SRBC as measured by the hemolytic plaque assay by up to 90%. This suppression was reversed by addition of rabbit anti-amyloid protein AA. Addition of SAA 24 hr or longer after initiation of cultures failed to give suppression, showing that the suppression by SAA occurs in the early recognition phase of the in vitro antibody response. Extensive washing of spleen cells reversed the suppression, suggesting an effect at the level of the cell membrane. A lack of suppression of the response to DNP-Ficoll (a T-independent antigen) indicated that there was no direct effect of SAA on B lymphocytes.

Purified SAA (100 µg/ml) isolated from the serum of casein-treated mice gave 95% suppression of SRBC response, and concentrations as low as 10 µg/ml gave significant suppression. This is much lower than the serum levels of SAA attained in humans and experimental animals after antigen challenge or immunization. It is suggested that a normal function of SAA may be to regulate immune response to certain foreign antigens or altered autologous antigens.
Slow Reacting Substance (SRS) from Ionophore A23187-Stimulated Peritoneal Mast Cells of the Normal Rat
Authors: Yecies Lewis D. , Johnson Stephen M. , Wedner H. James , Parker Charles W.
Published: December 2022

The generation of slow reacting substance (SRS) from ionophore A23187-stimulated rat peritoneal mast cells was enhanced by arachidonic acid (AA). This SRS generation was inhibited by 5,8,11,14-eicosatetraynoic acid (ETYA), an acetylenic analogue of AA and an inhibitor of both fatty acid cyclooxygenase and lipoxygenase. Indomethacin, a fatty acid cyclooxygenase inhibitor, had an enhancing effect upon SRS generation. This suggests SRS generation occurred through an ETYA sensitive step—perhaps a lipoxygenase. Radiolabel from [14C]-AA was incorporated into SRS with comigration of radio-activity and bioreactivity in silicic acid and thin layer chromatographies. Upon silicic acid chromatography, the active principle was eluted in the methanol fraction. Two-dimensional thin layer chromatography revealed chromatographic separation from other known spasmogenic substances and phospholipids. Mast cell SRS was found to display physiochemical properties similar to those of rat basophilic leukemia cell SRS, namely: that mast cell SRS generation was 1) enhanced by arachidonic acid; 2) inhibited by ETYA but not by indomethacin; 3) incorporation of [14C]-AA into the active principle; and 4) similar behavior during purification in silicic acid and thin layer chromatographies.
IgM Rheumatoid Factors in Mice Injected with Bacterial Lipopolysaccharides
Authors: Izui Shozo , Eisenberg Robert A. , Dixon Frank J.
Published: December 2022

Bacterial lipopolysaccharides (LPS) induced the formation of IgM rheumatoid factors (RF) in several strains of mice including athymic C57BL/6 nude mice, but not in the LPS-resistant C3H/HeJ mice. The RF induced by LPS reacted not only with murine IgG but also with IgG from cows, goats, guinea pigs, and humans. The kinetics of this RF response to injection of LPS were similar to those of antibody response against DNA and a hapten, dinitrophenyl (DNP), and to those of total IgM production. In addition, the RF activity of individual serum samples correlated significantly with levels of anti-DNA and anti-DNP antibodies and of IgM. Therefore, it is concluded that the induction of RF results from polyclonal antibody synthesis by B cells stimulated with LPS. This observation suggests that LPS or LPS-like substances may help to generate RF in patients with rheumatoid arthritis or with some infectious diseases.
The “Triggering” of Cytotoxic Cell Differentiation in Secondary Cultures by Subcellular Antigens
Authors: Henney Christopher S.
Published: December 2022

Sir: A number of investigators have recently reported that cell membranes, or solubilized proteins derived from them, are capable of inducing cytotoxic responses in secondary cultures (1–6). They have implied thereby that they are gaining insight into the specificity of the structures recognized by cytotoxic T cells.

An alternative interpretation seems equally plausible: that the membrane preparations act not directly on cytotoxic memory cells, but rather on helper T cells, which, by secreting soluble mediators, “trigger” cytotoxic cell differentiation. This interpretation is supported by several recent reports. Thus, Ryser et al. (7), Wagner and Rollinghoff (8), and this laboratory (Okada et al., in press) have shown that cell mediators secreted by primed Lyt I+ T cells can cause the direct differentiation of cytotoxic T cells in secondary (memory) cultures. Moreover, the specificity of the cytotoxic cells induced by these mediators is that of the antigen initially used for priming.
Cross-Species Contamination of Commercial Serum Proteins
Authors: Scharfstein Julio , Nussenzweig Victor
Published: December 2022

Dear Sir: After being plagued for some time by uninterpretable results in our radioimmunoassays, we began to suspect that the source of error could be the presence of bovine albumin in the preparations of commercial rabbit γ-globulin used in controls. Indeed, this unlikely contaminant was easily demonstrated by crossed immunoelectrophoresis and double immune diffusion in agarose in five out of five lots of rabbit γ-globulin, in one out of two preparations of rabbit albumin, and in the single sample of mouse immunoglobulin that was tested.1 Furthermore, we isolated bovine serum albumin from commercial rabbit immunoglobulin by anion exchange chromatography. By rocket immunoelectrophoresis we estimated the degree of contamination to be between 0.2 and 0.5%.

Sixteen years ago it was reported in Science (142: 1170, 1963) that bovine albumin could be detected in some lots of commercial rabbit albumin.
Beyond Cellular Immunology
Authors: Talmage David W.
Published: December 2022

Over the past 20 years we have seen the explosive development of a new scientific discipline, cellular immunology. At least 60% of the articles in our Journal and more than 60% of the allergy and immunology grants received or awarded by the National Institute of Allergy and Infectious Diseases in the past year involve experiments with lymphocytes and macrophages. This would have amused and pleased Dr. Gerald B. Webb, (Fig. 1), the first president of our Society. Dr. Webb was an early advocate of cellular immunology, but in his day this approach to immunity was not very popular; the humoral immunologists and serologists had the upper hand.

In the next few minutes I plan to review for you the origin and growth of cellular immunology and show how the very factors that led to its explosive development should be instrumental in creating a still newer discipline.

First a note about the origin of the word immunology.
Role of Lipid Fatty Acyl Composition and Membrane Fluidity in the Resistance of Acholeplasma Laidlawii to Complement-Mediated Killing
Authors: Dahl Jean S. , Dahl Charles E. , Levine R. P.
Published: December 2022

Acholeplasma laidlawii wild type and a mutant strain, comr-328, characterized by its resistance to complement-mediated killing, were examined for the effects of variations in their lipid fatty acyl moieties on membrane fluidity and the susceptibility of the cells to lysis by complement. Both cell types were cultured in media containing stearate, palmitate, or an equimolar mixture of palmitate and oleate. When stearate or palmitate served as the major membrane fatty acyl constituent, the sensitivity of the cells to complement-mediated killing via both the classical complement pathway and reactive lysis was increased, whereas the fluidity of the respective cell membranes was decreased. By comparison, when oleate was a major membrane fatty acyl moiety along with palmitate, the cell's sensitivity to complement-mediated killing decreased, whereas membrane fluidity increased. This pattern was observed for both strains although it was more pronounced in the wild-type strain than in comr-328. Although there appears to be a correlation between membrane fatty acyl composition, membrane fluidity, and complement-mediated killing in A. laidlawii, this correlation is obscure since the presence of cholesterol in the membrane, although affecting membrane fluidity, apparently does not alter the response of wild-type cells to lysis via the classical pathway.
Inhibition of the Mixed Lymphocyte Reaction by Fractionated Niridazole Urine Dialysate
Authors: Jones Brian M. , Bird Michelle , Griffin Peter J. A. , Massey Peter R. , Millar Douglas A. , Miller Julian J. , Reeves Suzanne , Salaman John R.
Published: December 2022

Urine dialysate from rats treated orally with 25 mg/Kg 3H-labeled niridazole was fractionated by DEAE-Sepharose column chromatography and was found to contain three radioactive metabolites and no parent compound. When human niridazole urine dialysate (NUD) was fractionated under identical conditions, fractions corresponding to the three rat NUD metabolites were found to inhibit the human one-way MLR. No inhibition was obtained with fractionated control urine dialysate. It was concluded that nonimmunosuppressive niridazole is metabolized by rats and man to produce three active compounds with the ability to suppress the in vitro response to alloantigens.
The Action of Strontium on Basophil Leukocytes and its Use to Probe the Relationship between Immunologic Stimulus and Secretory Response
Authors: Foreman John C. , Sobotka Anne K. , Lichtenstein Lawrence M.
Published: December 2022

Strontium will substitute for calcium in the activation of histamine secretion from human basophil leukocytes stimulated by an immunologic reaction or by the ionophore A23187. Strontium is required in 10-fold higher concentrations (1 to 10 mM) to activate histamine release compared with calcium (0.1 to 1.0 mM). In terms of maximum release obtainable for a particular immunologic stimulus, strontium is more effective than calcium. Results are presented to show that calcium and strontium act at the same site but strontium is a more sensitive probe for that site. Strontium can be used to demonstrate that immunologic stimuli activate calcium-binding sites in basophils even when no secretion is observed in the presence of calcium. It is suggested that the degree of secretion observed from basophils depends on the number of occupied Fc receptors for IgE and the coupling of these Fc receptors to calcium transport sites.