bluuebunny/crossref_metadata_2025 huggingface dataset:
Helper T Cell-Replacing Factors Secreted by Thymus-Derived Cells and Macrophages: Cellular Requirements for B Cell Activation and Synergistic Properties
Authors: Hoffmann M. K. , Watson J.
Published: December 2022

The biologic activities of helper T cell-replacing factors derived from concanavalin A-stimulated murine T cells (TRF-T) and from lipopolysaccharide-activated macrophages (TRF-M) have been compared. TRF-T stimulates immune responses to heterologous erythrocyte antigens (SRBC and BRBC) in T cell-depleted spleen cultures but not in macrophage-depleted spleen cultures. TRF-M stimulates immune responses in both T cell-depleted and macrophage-depleted spleen cultures. Under conditions where LPS stimulates the release of TRF-M from cultures of activated macrophages, TRF-T has no effect on TRF-M production. Thus, TRF-T does not appear to function by stimulating the release of TRF-M from macrophages. In macrophage-depleted spleen cultures, saturating concentrations of TRF-T and TRF-M when mixed together exhibit striking synergistic effects on the induction of immune responses to erythrocyte antigens. The kinetics of the synergistic effects of TRF-M and TRF-T are consistent with an effect of TRF-M on the production of TRF-T sensitive B cells.
Partial Purification and Characterization of Schistosoma Mansoni Soluble Egg Antigen with Con A-Sepharose Chromatography
Authors: Carter Clint E. , Colley Daniel G.
Published: December 2022

A crude preparation of Schistosoma mansoni soluble egg antigen (SEA) was subjected to affinity chromatography with concanavalin A (Con A) bound to Sepharose 4B. The resulting Con A fractions (bound and unbound) were characterized with sodium dodecyl sulfate (SDS) gel electrophoresis, immunoelectrophoresis, immunodiffusion, and lymphocyte blastogenesis techniques. In the fraction that did not bind to Con A there were at least two distinct antigens, and there were also at least two distinct antigens in the fractions that did bind to Con A. With SDS polyacrylamide gel electrophoresis, at least 20 distinct protein bands (Coomassie blue staining) and three glycoprotein bands (PAS reactive) were present in the unbound fractions from Con A chromatography. The bound fractions separated into at least six distinct glycoproteins with SDS electrophoresis. Although both the bound and unbound fractions contained precipitating antigens, only the bound fractions were capable of eliciting lymphocyte blastogenic responses.
Cyclical Changes in Susceptibility of a Myeloma Tumor (LPC-1) to Immune Destruction
Authors: Celis Esteban , Chang Tse Wen , Eisen Herman N.
Published: December 2022

Previous studies showed that when LPC-1 myeloma cells were grown as ascites tumor cells in syngeneic (BALB/c) mice, the cells harvested after 2 to 4 days of growth (“early” cells) were susceptible to lysis by cytotoxic T lymphocytes (CTL) and highly reactive with anti-H-2d antisera. Cells harvested after 12 to 14 days of growth (“late” cells) were resistant to lysis by CTL and poorly reactive with anti-H-2d antisera. The present study shows that exposure to trypsin or chymotrypsin or subtilisin (but not to thrombin or Staphylococcus A protease) promptly converts LPC-1 cells with the late phenotype into cells with the early phenotype. Comparison of radiolabeled cell surface proteins by gel electrophoresis showed that the late cells possess a prominent trypsin-sensitive, high m.w. (160,000 daltons) surface glycoprotein that is present in smaller amounts on the early LPC-1 cells. This glycoprotein (gp160) was not detectable in four other BALB/c tumors that do not undergo the early-late transition of LPC-1. That gp160 was produced by LPC-1 cells, rather than adsorbed by these cells as they grow in vivo, was evident from the presence of an indistinguishable metabolically labeled glycoprotein on cultured LPC-1 cells.
Receptor Cross-Linking and Histamine Release
Authors: DeLisi Charles , Siraganian Reuben P.
Published: December 2022

We develop and apply a mathematical method for relating the number of antigen-induced IgE clusters on the basophil plasma membrane to the amount of histamine released. The method is valid at low doses of antigen where, it is shown, the number of bound antigens and the number of IgE clusters are linearly proportional to the total concentration of antigen. Cross-linking of receptors on some portion of the surface is considered to be a necessary condition for transmission of a signal that leads to degranulation. No assumption is made about whether the degranulation thus stimulated is local or global. Analysis of the low-dose portion of the dose-response curve, however, indicates the following: 1) Degranulation is local, and the maximum number of local centers per cell (“compartments”) is small (≤ 10). 2) The number of IgE clusters required to initiate such local release is small but variable and depends upon the cell source and antigen valence. The results varied between three and eight for Con A, AgE, and anti-IgE. 3) These numbers represent thresholds for release; i.e., to a good approximation some minimum number of antigens in excess of one must be multiply bound before any histamine can be released. This suggests a high degree of cooperativity in the biochemical sequelae to clustering. 4) Only a small fraction of clusters are transmitting a release signal. This suggests, and is consistent with the idea, that clusters are interacting with some other membrane molecule and that only those that are bound to the other molecule are biologically active. 5) With respect to transmitting a signal for histamine release the number of independent clusters appears to be a more fundamental quantity than the total number of cross-linked receptors; i.e., there is little difference in activity as the size of a cluster increases due to increase in ligand valence.
Receptor Cross-Linking and Histamine Release
Authors: Delisi Charles , Siraganian Reuben P.
Published: December 2022

We report the results and analysis of dose-response and desensitization studies of rabbit basophils by a variety of antigens. Unlike the response of human basophils, the histamine release curves rarely rise and fall in a symmetric fashion, but more often either rise to a plateau, peak, and then fall to a non-zero plateau, or exhibit bimodality. In those cases in which the curves peak, preincubation with antigen in the absence of Ca++ has a smaller effect at the high and low concentration portions of the curve than it does toward the center, where the inhibitory effect is maximum.

We show that the results can be explained by a simple model in which: 1) The signals for release and desensitization both increase as the concentration of receptor cross-links increases and 2) the signals reach a plateau (i.e., they have a maximum value) when the number of receptor cross-links becomes sufficiently large. By changing the parameters of the model, the balance between desensitization and release changes, and all the observed patterns can be reproduced. In particular, if both signals saturate (reach their maximum value) at a ligand concentration below that of maximum cross-linking, the histamine release curve will plateau. If the release signal saturates but the inhibitory signal continues to increase as ligand concentration increases toward its optimum value, the histamine release curve will be bimodal.

Many of the parameters can be identified with those that affect receptor clustering. These include the affinity of IgE for the antigen, the number of IgE molecules per basophil, IgE hinge flexibility, membrane fluidity, and antigen concentration and valence. This identification permits a variety of additional predictions. One such prediction is that the difference between a dose-response curve that plateaus and one that is biphasic and unimodal reflects easier desensitization of cells responsible for the latter curve and/or that have a smaller number of cross-links. Both predictions are readily testable experimentally.
Genetic and Biochemical Evidence for the Involvement of a Bacterial Component in the Mitogenic Properties of Polyribonucleotides on Murine B Lymphocytes
Authors: Kelly Kathleen , Watson James
Published: December 2022

The mitogenic response of C3H/HeJ mice to the B cell mitogens, poly C and poly I, is approximately one-half the response measured in various LPS-responder strains. C3H/HeJ mice respond normally to poly I:C, the heteroduplex polymer. The low responder phenotype of C3H/HeJ mice to poly C and poly I is shown by an analysis of (C3H/HeJ × C57BL/6J-By-Ps)F1 × C3H/HeJ backcross progeny to result from a gene locus that is closely linked or identical to the defective LPS response locus expressed by the C3H/HeJ strain. The entire mitogenic activity in poly C preparations and most of the mitogenic activity in poly I preparations is insensitive to ribonuclease degradation. Hot aqueous phenol extraction of the polynucleotides separates the majority of the mitogenic activity that is soluble in the combined interface and phenol phase fraction from the aqueous soluble polynucleotides. The ribonuclease-insensitive, phenolsoluble contaminant elicits a reduced response in C3H/HeJ mice as compared to an LPS responder strain. We conclude that 1) poly C has no inherent mitogenic activity; 2) poly I preparations contain both ribonuclease-sensitive and insensitive mitogenic activities; 3) the ribonuclease-resistant mitogenic activity in polynucleotide preparations has physical properties unlike those of LPS or lipid A; and 4) the product of the LPS response gene has an effect upon the mitogenic stimulation of spleen cells by the contaminant.
Patterns of Immune Reactivity in Autoimmune Murine Strains
Authors: Theofilopoulos Argyrios N. , Shawler Daniel L. , Katz David H. , Dixon Frank J.
Published: December 2022

We compared spleen cells from immunologically normal mice to those from five autoimmune murine strains (NZB, NZB × W, BXSB, MRL/l, MRL/n) in the cell-mediated lympholytic response in vitro to spleen cells from syngeneic, allogeneic but H-2 identical and H-2 incompatible mice. NZB spleen cells sensitized with irradiated allogeneic but H-2d identical spleen cells lysed unmodified H-2 identical cells in a strong, cross-reactive, unidirectional response, and lysed similarly, but to a lesser extent, syngeneic cells. The NZB anti-H-2d activity was mediated by T cells and was directed against both T and B target cells. Although the anti-H-2d activity is known to be directed against minor histocompatibility differences, it appears from our experiments with congenic mice that the activity is to some extent recognized in the context of the H-2 haplotype, since it was optimum when the NZB cells matched the target cells at the K or D end of H-2.

NZB cells sensitized in vitro by H-2 identical cells were only minimally stimulated to proliferate; however, such cells were able to induce in vivo GVH reactions in NZB mice. Similar in vitro anti-H-2d reactivity was observed with NZB × W spleen cells. In contrast, spleen cells from BXSB, MRL/1, and MRL/n mice were not sensitized in vitro with H-2 identical cells. Spleen cells from all autoimmune murine strains incubated with irradiated syngeneic cells were devoid of any cytolytic activity against syngeneic target cells. Cells from all autoimmune strains sampled during youth exhibited a normal effector function against H-2 incompatible spleen cells, and this function declined moderately with age and appearance of disease. In contrast to all other autoimmune strains, cells from MRL/1 mice were inadequate stimulators with H-2 incompatible cells, possibly due to massive T cell lymphoproliferation seen in these mice. Although the observed anti-H-2d activity of cells from New Zealand mice (NZB, NZB × W) may play a role in the development of their disease, the possibility that such a mechanism serves as a common etiologic pathway in the development of the murine SLE-like syndromes seems unlikely since the other autoimmune strains studied were devoid of such activity.
Antibody Blockade of Lysis by T Lymphocyte Effectors Generated against Syngeneic SV40 Transformed Cells
Authors: Gooding Linda R.
Published: December 2022

Antibody blockade of cell-mediated lysis was used to probe the cell surface structures recognized by T lymphocytes generated to syngeneic SV40 virus-transformed mouse cells. Alloantisera to H-2 antigens were highly effective in inhibiting lysis. Anti-H-2 antibody blockade of lysis was haplotype specific even on transformed F1 target cells. Inhibition occurred only when antibody was bound to the target cell. Antibody binding to the effector did not inhibit lysis. Inhibition required that antibody be bound to the H-2 molecule itself; antibody to molecules associated with H-2, such as β2-microglobulin, had no effect.

Attempts to block lysis by using antisera to known virus-coded molecules were uniformly unsuccessful. These results are discussed in light of current controversy concerning the nature of the SV40 virus-specific transplantation antigen.
Evaluation of the Role of C4 in the Cellular Immune Response in Vitro
Authors: Burger Reinhard , Shevach Ethan M.
Published: December 2022

The demonstration that antisera to human C4 inhibit the human mixed lymphocyte reaction and the response to the mitogen, phytohemagglutin, suggested a possible role of C4 in the afferent phase of immune recognition. An attempt was made to reproduce and extend these observations in the guinea pig. Monospecific antisera to C4 were raised in C4-deficient guinea pigs, rabbit, or goat. The antisera were tested for inhibition of the unidirectional mixed leukocyte reactions between strain 13 and strain 2 guinea pigs, for inhibition of proliferation induced by the mitogens, phytohemagglutinin and concanavalin A, and for inhibition of the proliferative response of immune T lymphocytes to soluble protein antigens. In a large number of experiments no reproducible inhibition was found when the various cultures were performed in the presence of antiserum to C4. Furthermore, lymphocytes from C4-deficient guinea pigs responded as well as lymphocytes from strain 13 animals to specific antigen or in mitogen-induced proliferation, indicating a normal capacity of T cells from C4-deficient animals for recognition of antigen or mitogen and for proliferation. Antigen- or mitogen-pulsed macrophages from C4-deficient guinea pigs were able to effectively stimulate T cell proliferation, suggesting a normal capability for antigen or mitogen presentation. These data argue against a possible role for C4 in antigen, mitogen, or alloantigen-induced T lymphocyte activation and indicate that the demonstrated functional association of C4 and MHC-mediated reactions in the human does not generally apply to another species.
Macrophage-T Cell Interactions Involving Listeria Monocytogenes—Role of the H-2 Gene Complex
Authors: Farr Andrew G. , Kiely Jeanne-Marie , Unanue Emil R.
Published: December 2022

This study reports on the in vitro interactions between T cells from Listeria-immunized mice, macrophages from normal mice, and heat-killed Listeria organisms. This interaction was assayed either by determining the amount of a thymocyte mitogen in culture fluids after 24 hr, or by estimating the degree of T cell proliferation after 96 hr. Each assay depended on critical concentrations of macrophages, T cells, and heat-killed Listeria, points that were evaluated in a number of experiments. Both assays required specific Listeria-immune T cells. For an effective interaction, the T cells and the macrophages had to share the I-A region of the H-2 gene complex. Macrophages bearing Ia, which represented a minor population of macrophages, were essential for the proliferative response to macrophage-associated Listeria. Also, Ia-bearing macrophages were an important component in the interactions leading to increased secretion of mitogen. The immunogenic moiety associated with Listeria was short-lived, disappearing 24 hr after uptake of Listeria by macrophages. The interactions were not blocked by anti-Listeria antibodies but were partially sensitive to trypsinization.
Tumor-Basophil Interactions in Vitro—A Scanning and Transmission Electron Microscopic Study
Authors: Dvorak Ann M. , Galli Stephen J. , Galli Anne S. , Hammond M. Elizabeth , Churchill W. Hallowell , Dvorak Harold F.
Published: December 2022

Purified guinea pig basophils, or basophils either specifically degranulated with antigen or nonspecifically degranulated with lectin, were cultured with guinea pig line 1 hepatoma cells for 1 to 24 hr and studied ultrastructurally. As early as 1 hr of culture, degranulated or nongranulated basophils and tumor cells formed close contacts by mutually intertwined elongated cell processes and also in cultures containing degranulated basophils, extruded membrane-free basophil cytoplasmic granules became firmly attached to tumor cells. At later intervals, some tumor cells cultured with basophils exhibited cytostatic and cytopathic changes, including dense mitochondria, centralization of organelles, dilated perinuclear and rough endoplasmic cisternae, cell swelling and cytoplasmic lucency, disrupted cytoplasmic organelle and plasma membranes, nuclear pyknosis and fragmentation. Some tumor cell specialized surface attachments were either disrupted or damaged at points of basophil or basophil granule adhesion. Tumor damage was most extensive in cultures containing degranulated basophils, although only a minority of tumor cells (<10%) was affected. Tumor injury was seen much less frequently in the presence of nondegranulated basophils, and was absent in control cultures of tumor alone. The occasional viable tumor cells that phagocytosed basophil granules were apparently unharmed, suggesting that internalization of basophil granules by tumor cells is not cytotoxic.
Lymphocytes Bearing Fc Receptors for IgE
Authors: Yodoi Junji , Ishizaka Kimishige
Published: December 2022

Human peripheral blood lymphocytes from ragweed-sensitive individuals were fractionated by a rosette technique with neuraminidase-treated sheep erythrocytes (En). A significant number of FcεR-bearing cells were detected in the T cell fraction by rosette formation with ox erythrocytes coated with human IgE (E'-hIgE), or by the binding of fluoresceinated F(ab′)2 fragments of anti-IgE complexed with IgE. The majority of these lymphocytes in the T cell fraction formed mixed rosettes with En and E'-hIgE. T cells bearing FcεR were demonstrated in the mesenteric lymph node cells from rats infected with Nippostrongylus brasiliensis (Nb); about 3% of the total cells in a B-depleted fraction formed rosettes with ox erythrocytes coated with rat IgE (E'-rIgE). Most of the rosette-forming cells (RFC) in the fraction were stained by antibodies specific for rat T cells in immunofluorescence. A substantial proportion (5 to 10%) of lymphocytes in rat-activated T cells, which were prepared by transfer of Lewis strain thymocytes to Sprague-Dawley rats, formed rosettes with E'-rIgE, and essentially all of the RFC were stained by anti-T cell antibodies. The results indicated that a subset of human and rat T cells possess FcεR.

Evidence was obtained that FcεR on T cells are distinct from Fc receptors for IgG (FcγR), which were demonstrated by rosette formation with ox erythrocytes coated with rabbit IgG antibodies (EA). Mixed rosette assays showed that more than a half of FcεR-bearing human T cells failed to bind EA. Similarly, the majority of FcεR-bearing T cells in the lymph nodes of Nb-infected rats failed to bind EA. In activated T cell preparations, however, about three-fourths of FcεR-bearing cells formed mixed rosettes with EA, indicating that some T cells bear both FcεR and FcγR.
Accessory Cell Requirements for the Generation of Cytolytic T Lymphocytes
Authors: Lutz Charles T. , Fitch Frank W.
Published: December 2022

A system is presented that may simplify the study of accessory cell requirements for CTL generation. Cortisone resistant (CR) thymocytes containing alloreactive CTL precursors do not respond to allogeneic tumor cells unless non-T accessory cells are added to culture. In addition, splenic T cells do not respond to allogeneic tumor cells in the absence of non-T accessory cells. These accessory cells share several properties of macrophages.
Cytotoxic Thy 1.2-Positive Blasts with NK-Like Target Selectivity in Murine Mixed Lymphocyte Cultures
Authors: Kärre Klas , Seeley Janet K.
Published: December 2022

CBA × A/Sn spleen cells kept in H-2b allosensitized and control cultures (all containing 10% FCS) were assayed for cytotoxic activity in the 51Cr-release assay against YAC-1, an A/Sn-derived lymphoma that is highly sensitive to murine NK cells. After a decline during the first 24 hr of culture, anti-YAC-1 activity reappeared with a peak on day 3, thereafter it decreased to baseline levels as the allospecific activity (against RBL-5 H-2b lymphoma cells) peaked on day 5. This early, transient, anti-YAC-1 activity, termed “anomalous killing (AK)”, also occurred in control cultures, but was weaker and less reproducible there. With effectors from day 3 MLC, there was no cross-competition between the AK and the allospecific component in cold target inhibition experiments. The same effectors lysed human and murine semisyngeneic tumors in the same preferential order as fresh NK cells. The cells responsible for AK were nonadherent, nonphagocytic, of large size (sedimentation velocity >6 mm/hr), and sensitive to treatment with monoclonal anti-Thy 1.2 antibodies + complement. In contrast, NK activity of fresh spleen cells was mediated by small lymphocytes and was not decreased by anti-Thy 1.2 treatment in parallel experiments. The AK may represent further differentiation of NK cells or activation of a mature T cell subset. The mechanism of activation in relation to the in vivo immune response and in vitro induced NK-like activity in human systems is discussed.
Modulation of Stimulated Phospholipid Metabolism in Mast Cells by Pharmacologic Agents That Increase Cyclic 3′,5′ Adenosine Monophosphate Levels
Authors: Kennerly Donald A. , Secosan Craig J. , Parker Charles W. , Sullivan Timothy J.
Published: December 2022

Phospholipid (PL) metabolism is markedly stimulated during mediator release from isolated rat mast cells. The present studies were performed to extend studies of the association of these PL reactions with the mediator release process and to probe the mechanisms by which pharmacologic agents that increase cyclic 3′,5′ adenosine monophosphate (cAMP) levels inhibit mediator release. Theophylline (1 to 20 mM) and N6, O2-dibutyryl cAMP (DBcAMP, 1 to 10 mM) markedly inhibited 32PO4 incorporation into phosphatidic acid (PA), phosphatidylinositol (PI), and phosphatidylcholine (PC) in unstimulated mast cells. Epinephrine (1 to 100 µM) had no effect on PL labeling in resting cells. The histamine release and increased 32PO4 labeling of PA, PI, and PC observed in mast cells stimulated by a goat anti-rat IgE antiserum were inhibited in parallel by theophylline (ID50 approximately 5 mM for all parameters) and by DBcAMP (ID50 approximately 3 mM for all parameters). Epinephrine (1 to 100 µM) did not modify mediator release or labeling of PA or PC, but did modestly inhibit PI labeling (23% inhibition at 100 µM epinephrine). Theophylline (above 3 mM) was found to arrest significantly ongoing mediator release and PL labeling when added as late as 2 to 5 min after anti-IgE challenge. The parallel effects of these pharmacologic interventions on mediator release and PL metabolism provide further evidence that the two processes are biochemically linked.
Expansion of Human Lymphocyte Populations Expressing Specific Immune Reactivities
Authors: Malissen Bernard , Charmot Dominique , Liabeuf Andre , Mawas Claude
Published: December 2022

Fourteen-day-primary MLR cultures, containing cells positively selected for the alloantigens of the stimulator, express the following immune reactivities: accelerated proliferation toward the specific stimulator or any allogeneic cell sharing with it HLA-D/DR antigens (PLT), strong specific cytotoxicity toward the stimulator cells (CML) or any cell sharing MHC products with it, and a strong inhibitory effect upon the proliferation in a fresh MLR of the original combination. Such primed cells can still proliferate when exposed to mitogenic doses of lectins. We have investigated whether, after a cycle of proliferation induced by mitogens, these cells could still express a specific immune reactivity such as CML. PHA, Con A, and PKW all induce a strong proliferation in primed cells, but only those primed cells stimulated by PKW were specifically cytotoxic. In contrast PHA- and Con A-stimulated cells had lost their lytic function. The absence of cytolytic function in primed cells stimulated by PHA is a complex phenomenon. Experiments where day 7 MLR cells were separated by 1 × G velocity sedimentation into blasts and small cells suggest a mitogen-dependent inhibitory effect upon CTL differentiation from day 7 blast cells, which requires cells that were small on day 7.

We have also observed a direct effect of PHA upon differentiated CTL: CTL incubated for as few as 2.5 hr in the presence of PHA lost 80% of their lytic potential, whether the incubation was done at 37 or 4°C. This effect was not seen after incubation with PKW. The effect of PHA is reversible: CTL function can be rescued provided the effector cells are cultured for at least 24 hr in the absence of PHA (with or without additional stimulator cells). In view of this reversible effect of PHA upon the expression of the lytic function by primed cells, we conclude that both PHA and PKW induce proliferation of the CTL clones included within the primed population, although after PHA (or Con A) stimulation the lytic function is not readily detectable. We propose provisionally to call the inhibitory effect of PHA and Con A an “eclipse” of the lytic function of primed cells.
Degranulation of Human Basophils: Quantitative Analysis of Histamine Release and Desensitization, Due to a Bivalent Penicilloyl Hapten
Authors: Dembo Micah , Goldstein Byron , Sobotka Anne K. , Lichtenstein Lawrence M.
Published: December 2022

We present a quantitative model of histamine release from human basophils due to the symmetric bivalent hapten, Bis benzylpencilloyl 1,6 diaminohexane, (BPO)2. The major elements of model are: 1) The histamine content of basophils is divided into a large number of discrete and quasi-independent “quanta.” 2) A certain fraction of the quanta in a population of basophils are nonreleasable. 3) Release of the contents of a quanta is a stochastic event. 4) The binding and cross-linking reaction between (BPO)2 and cell surface IgE approaches thermal equilibrium on a time scale that is very short compared to the time scale of histamine release from or desensitization of basophils. 5) The probability per unit time of the release event, PR, is a function of the extracellular calcium concentration [Ca]ex, and of the number of cross-linked IgE molecules per cell, Xpoly. In addition, the functional dependence of PR on Xpoly for fixed [Ca]ex is sigmoid, and the dependence of PR on [Ca]ex is such that PR → 0 as [Ca]ex → 0 for all values of Xpoly. 6) Antigen-specific IgE molecules and/or their associated Fc receptors are inactivated at a rate that is a saturable function of Xpoly. Furthermore, this process of inactivation is not dependent on the extracellular calcium concentration.

The model is tested by carrying out a detailed fitting to histamine release data obtained by using cells from a single donor. Good agreement between theory and experiment is obtained, and all the parameters of the model are determined. The ways in which the model can be extended to include systems more complex than the (BPO)2 system are briefly discussed.
Murine B Cell Leukemia (BCL1): Organ Distribution and Kinetics of Growth as Determined by Fluorescence Analysis with an Anti-Idiotypic Antibody
Authors: Krolick K. A. , Isakson P. C. , Uhr J. W. , Vitetta E. S.
Published: December 2022

The life history of a transplantable B cell leukemia (BCL1) that arose spontaneously in a BALB/c mouse is described. Animals bearing this tumor live from 2 to 4 months in apparently good health despite massive splenomegaly and leukemia. Antibody to the idiotype or λ light chain of the tumor IgM was used in conjunction with the fluorescence-activated cell sorter to identify tumor cells in the BCL1-bearing mice. The results suggest that these cells multiply and differentiate in the spleen and subsequently emigrate to the blood. Tumor cells do not recirculate as evidenced by their failure to enter the thoracic duct or to infiltrate lymph nodes to a significant extent. During tumor growth, a population of T cell blasts appears that may be involved with an immune response against the tumor.
Radiolabeling and Isolation of Thy-1 Active Glycolipids from Murine Brain and Lymphoma Cell Lines
Authors: Kato K. P. , Wang T. J. , Esselman W. J.
Published: December 2022

Murine Thy-1 active glycolipids were isolated and characterized by radiolabeling, thin layer chromatography (TLC), and an immune response-plaque forming cell (PFC) assay. Brain gangliosides were labeled in vivo by intracranial injection of [1-14C]N-acetylmannosamine (ManNAc). Gangliosides were extracted, separated by one and two dimensional TLC, and visualized by autoradiography. The gangliosides isolated from AKR (Thy-1.1) and ICR (Thy-1.2) mice were assayed for Thy-1.1 and 1.2 activity with an anti-Thy-1 PFC assay. One Thy-1.1 active compound was identified out of about 15 AKR gangliosides. None of the AKR gangliosides had Thy-1.2 activity in the PFC assay. Similarly, one Thy-1.2 active ganglioside was identified out of the ICR gangliosides and none of these had Thy-1.1 activity. Thus the brain gangliosides demonstrated allogeneic specificity for Thy-1 antigen.

BW5147 (Thy-1.1) and S49.1 (Thy-1.2) murine lymphoma cell lines that express Thy-1 were also examined for Thy-1 active glycolipids. The cell lines were incubated in vitro with either [1-14C]palmitate or [1-14C]-ManNAc, and the glycolipids were extracted, separated by two dimensional TLC, and visualized by autoradiography. A Thy-1.2 active compound was identified from the S49.1 cells. None of the BW5147 glycolipids had Thy-1.2 activity and none of the S49.1 glycolipids had Thy-1.1 activity. Neuraminidase and mild HC1 treatment of Thy-1 active glycolipids abrogated the subsequent PFC response. The Thy-1 glycolipids bound to DEAE cellulose as expected for gangliosides. We therefore conclude that Thy-1 antigenicity is associated with gangliosides of mouse brain and lymphoma cells.
Inhibition of Growth of ASL-1w Murine Leukemia Cells by Anti-Thymus Leukemia Antigen (TL) Serum in the Absence of Complement
Authors: Wolcott Michael , Pickering Jerry W. , Williams Jackie L.
Published: December 2022

Previous studies have shown that the cell surface expression of thymus leukemia antigen (TL) on ASL-1w leukemia cells varies with the progression of the cells through the growth cycle. Expression of TL is maximal in S phase, and its quantitative expression varies directly with DNA synthesis. In the present study, the effect of anti-TL serum on the growth of ASL-1w cells was examined. The antiserum, tested in the absence of complement, affected the growth of these cells in biphasic manner. When the antiserum concentration was 0.1% or greater, there was a rapid decline in the rate of DNA synthesis, and after 5 to 7 hr, cell death. When the antiserum concentration was 0.067% or less, the decline in the rate of synthesis of DNA did not become apparent until 5 to 6 hr after treatment. Under these conditions, there was approximately a 20% increase in cell number after 24 hr of culture. The hypothesis that treatment of ASL-1w cells with the lesser concentration of anti-TL serum blocks the cells in G2 phase of the cell cycle is discussed.
Lymphocytes Bearing Receptors for IgE
Authors: Yodoi Junji , Ishizaka Kimishige
Published: December 2022

Culture of normal rat lymphocytes with rat IgE at 37°C resulted in an increase in the proportion of FcεR(+) cells. In agreement with this finding, the ability of normal lymphocytes to bind 125I-IgE markedly increased after overnight incubation with IgE. Evidence was obtained that FcεR(+) cells were derived from FcγR(+) cells. FcεR(+) cells were not induced in vitro if FcγR(+) cells were depleted from normal lymphocytes before incubation with IgE. Kinetic studies for the induction of FcεR(+) cells showed that the majority of FcεR(+) cells induced in vitro bore FcγR at the early stage of differentiation. The FcεR-FcγR double-bearing cells converted to FcεR signal-bearing cells after further incubation with IgE. It was also found that induction of FcεR(+) cells by IgE was inhibited by rabbit IgG (RGG) or soluble antigen—IgG antibody complexes that have affinity for FcγR, but FcεR induced in vitro were specific for IgE. The results suggested that the binding of IgE with FcγR provided a signal to form and/or express FcεR.

A high proportion of FcεR(+) cells in the mesenteric lymph nodes from rats infected with Nippostrongylus brasiliensis was maintained if the lymphocytes were cultured in IgE, whereas the proportion markedly diminished if the cells were kept overnight in IgE-free medium. Evidence was obtained that the maintenance in the proportion of FcεR(+) cells does not involve recruitment of FcγR(+) cells to FcεR(+) cells; the proportion of FcεR(+) cells in a FcγR-depleted fraction was maintained by IgE. Neither RGG nor antigen-IgG antibody complexes affected the maintenance of the FcεR(+) cells by IgE. The results collectively suggested that the binding of IgE with either FcγR or FcεR stimulate lymphocytes to form FcεR.
The Kinetics for Binding and Catabolism of Aggregated IgG by Rat Peritoneal Macrophages
Authors: Knutson David W. , Kijlstra Aize , van Es Leendert A.
Published: December 2022

Mononuclear phagocytes in the reticuloendothelial system (RES) bind and catabolize circulating antigen-antibody complexes. These functions were studied with an in vitro model to elucidate the cellular mechanisms and rate-limiting step for catabolism of soluble immune complexes by mononuclear phagocytes. The model consisted of reacting stable heat aggregates of 125I-IgG (A-IgG) (0.01 to 10 µg) at 37°C for 10 min to 24 hr with 1.0 × 106 adhering peritoneal macrophages from normal rats. A-IgG were then separated into cell-associated, acid-precipitable (unreacted) and acid-soluble catabolized fractions. With subsaturating doses of A-IgG, binding and catabolism of A-IgG followed the kinetics for consecutive first-order reactions with binding the ratelimiting step (binding rate constant = ka = 0.7 to 5.5 × 10-3 min-1; catabolism rate constant = kc = 20.0 to 55.0 × 10-3 min-1). At higher doses of A-IgG, the binding step became saturated (and ka fell) while catabolism remained first order (with constant kc). The maximum clearance velocity (Vmax) was a function of A-IgG size in that more micrograms but fewer molecules of A-IgG9 could be cleared per unit time than A-IgG74 (1200 A-IgG9 and 540 A-IgG74 molecules per cell per min). Most cell-associated A-IgG were present on the cell surface where they were accessible to release by trypsin. On the basis of these observations, it was postulated that binding becomes saturated because of limited availability of receptors for the Fc portion of IgG. Assuming that pinocytosis of A-IgG was necessary before A-IgG were catabolized, these data further suggest that pinocytosis of A-IgG is slower than the subsequent catabolic step and that bound A-IgG are pinocytosed at a constant fractional rate. This would seem to require mechanisms to concentrate A-IgG at certain sites on the membrane and then selectively to interiorize the A-IgG bearing portions.
Rabbit B Spleen Lymphocytes and T Helper Cells
Authors: Cavaillon J. M. , Udupa T. N. S. , Chou C-T. , Cinader B. , Haeffner-Cavaillon N. , Dubiski S.
Published: December 2022

The response to anti-allotype (anti-Ab4), Nocardia Water Soluble Mitogen (NWSM), pneumococcal polysaccharide type III (SSS III), and human Fc fragments of various purified and unfractionated rabbit spleen cell populations was determined in terms of 3H-thymidine uptake. B cells were isolated either from untreated suspensions of spleen cells or from suspensions from which adherent and phagocytic cells were removed. The purification factor was greater than the enhancement of 3H-thymidine uptake by anti-Ab4, NWSM, and SSS III as compared with the response of unfractionated spleen cells. It thus appears that a helper cell was involved: the mitogen response of purified B cells was enhanced by the addition of T cells. B subpopulations were separated by sedimentation or by rosetting, which allowed us to separate Fcγ receptor-bearing cells from cells that did not possess this receptor. There were differences between cells responding to B mitogens not only in sedimentation velocity but also in the absolute number of cells. B cells bearing the Fcγ receptor were less responsive to anti-Ab4 and more responsive to SSS III, NWSM, and human Fc than were B cells lacking the Fcγ receptor.
Autologous and Allogeneic Intercellular Interactions: Modulation by Adherent Cells, Irradiation, and in Vitro and in Vivo Corticosteroids
Authors: Katz Paul , Fauci Anthony S.
Published: December 2022

The present study investigated the intrinsic blastogenic reactivity within T and non-T lymphocyte populations as well as between these populations in the autologous and allogeneic mixed lymphocyte reactions (MLR). T cell-depleted lymphocytes exhibited after 6 days in culture, high levels of intrinsic blastogenic activity that were sensitive to irradiation and in vitro hydrocortisone. Depletion of adherent cells shifted the peak T cell-depleted intrinsic blastogenesis from 6 days in culture to 2 to 3 days after initiation of culture. T cellenriched cells displayed low intrinsic blastogenic responses that were relatively unaffected by irradiation and in vitro hydrocortisone. The removal of adherent cells from T cell-depleted suspensions (stimulators) augmented both the autologous and allogeneic MLR, whereas irradiation of cells had no effect on these reactions. In vitro hydrocortisone suppressed the autologous MLR and, to a much lesser extent, the allogeneic MLR. I.V. administration of hydrocortisone to normal volunteers suppressed the autologous MLR at 4 hr after injection. The responsiveness of T cells obtained at 4 hr to autologous and allogeneic non-T cell stimulation was markedly diminished. Non-T cells obtained 4 hr after i.v. hydrocortisone had greater stimulatory capacity for both the autologous and allogeneic MLR than did T cell-depleted cells obtained before hydrocortisone administration. These studies demonstrate that autologous and allogeneic intercellular interactions between lymphoid cells are susceptible to a variety of in vitro and in vivo influences and that such interactions may reflect relevant immunoregulatory events occurring in vivo.
H-2 Unrestricted Cytotoxic T Cell Activity against Antigens Controlled by Genes in the QA/TLA Region
Authors: Forman James
Published: December 2022

The specificity of H-2 unrestricted cytotoxic T cells was analyzed in secondary CML responses. A/J strain effector cells, sensitized against A.Tlab lymphoid cells, lysed target cells from strains with differing H-2 haplotypes but all sharing Qa-1b/Tlab alleles; whereas, target cells from strains with Qa-1a/Tlaa were not. When B6.Tlaa animals were in vivo-primed and challenged in vitro with B6 stimulator cells, no cytotoxic effector cell activity was generated. However, if B6.Tlaa animals were primed in vivo with A.BY cells and then rechallenged in vitro with either A.BY or B6 stimulator cells, cytotoxic effector cells were generated that lysed target cells from strains with Qa-1b/Tlab alleles. This suggests that factors in addition to Qa/Tla may play a role in the generation of anti-Qa/Tla effector cell activity. It was also noted that targets from strains with Qa-1a/Tlaa alleles were killed, although to a much lesser extent than the Qa-1b/Tlab targets. SWR anti-DBA/1 effector cells strongly lysed target cells from strains with Qa-1b/Tlab, lysed Qa-1a/Tlaa targets to a lesser extent, and produced no cytolytic effect on B6.Tlaa target cells. These data suggest that in addition to a CML target antigen associated with Qa-1b/Tlab, there may be an additional specificity recognized by cytotoxic T cells controlled by a gene outside of Qa-1b/Tlab.
Mechanisms of Idiotype Suppression
Authors: Kim Byung S.
Published: December 2022

Specific tolerance of BALB/c spleen cells to phosphorylcholine (PC) was induced in vitro by treatment with antibodies specific for the idiotype of TEPC-15 myeloma protein (anti-T15id) and a PC-containing antigen, R36a. The presence of anti-T15id antibody for at least 2 days was necessary to achieve a full level of suppression (>98%) of anti-PC production in the presence of R36a. The anti-PC response of a nylon wool-separated B cell population was also greatly suppressed (94%) only after a 48-hr treatment with anti-T15id antibody plus antigen. In contrast, either isolated B or unseparated spleen cells treated with anti-T15id alone (without antigen) were fully responsive to PC. Kinetic studies inducated that treatment of cells with anti-T15id antibodies either in the presence or absence of R36a resulted in a gradual suppression of anti-PC responses at a similar rate during an initial 12-hr period. However, further treatment (24 hr) of cells with anti-T15id antibody alone led to recovery of anti-PC response from the suppression, whereas incubation with the anti-idiotype antibody plus R36a induced further suppression. These results indicate that anti-idiotype antibodies can directly inactivate B cells, and this inactivation becomes irreversible only after a continuous exposure to the antibodies in the presence of specific antigen.
Regulation of Cell Mediated Immunity by Antibodies: Possible Role of Anti-Receptor Antibodies in the Regulation of Contact Sensitivity to DNFB in Mice
Authors: Sy Man-Sun , Moorhead John W. , Claman Henry N.
Published: December 2022

The duration of maximal contact sensitivity in the mouse is regulated, at least in part, by specific antibodies. Mice sensitized with optimal doses of 2,4-dinitrofluorobenzene (DNFB) develop maximum delayed hypersensitivity in 4 to 5 days as measured in vivo by ear swelling. The intensity of this reaction declines rapidly, and by 12 days post-sensitization significant immunity is no longer detectable. Serum taken from animals 9 to 15 days post-sensitization will block the ability of DNFB-immune lymph node (LN) cells to passively transfer immunity. The suppressive molecules are immunoglobulins, but they do not have anti-DNP activity and are not associated with DNP as antigen-antibody complexes. The suppressive serum is antigen specific, i.e., it blocks DNFB-immune LN cells but not TNCB- or oxazolone-immune LN cells, and it lacks strain specificity in inhibiting passive transfer. The suppressive serum can be adsorbed by DNFB-immune LN cells (both syngeneic and allogeneic), but not by normal LN cells. Furthermore, the suppressive activity of the serum is adsorbed by and can be recovered from an affinity column conjugated with purified mouse anti-DNP antibodies.

We conclude that blocking antibodies arise during sensitization with DNFB and down-regulate the duration of sensitivity. These antibodies are directed against surface components of immune LN cells. They possess anti-idiotype activity and represent part of the immunoregulatory elements within a complex network of anti-idiotype/idiotype interaction.
Antigenic Determinants of Influenza Virus Hemagglutinin
Authors: Jackson David C. , Brown Lorena E. , White David O. , Dopheide Theo A. A. , Ward Colin W.
Published: December 2022

Polypeptide fragments obtained by cyanogen bromide (CNBr) cleavage of the hemagglutinin (HA) of the A/Memphis/102/72 (H3N2) strain of influenza virus were inoculated into rabbits, and the resulting antisera were examined for the presence of antibodies capable of reacting with intact virus, detergent-solubilized hemagglutinin, hemagglutinin heavy chain (HA1), or CNBr fragments derived from HA. We have shown that it is possible to raise antibodies to individual fragments including those representing regions of the hemagglutinin molecule that are not normally antigenic within the intact virion. The antibodies are specific for the fragment to which they were raised. In those cases where the immunogen contains carbohydrate, a small proportion of the antibody is directed against these side chains, but this can be removed by adsorption with an unrelated influenza virus. Although only antibodies raised against intact HA are capable of neutralization of virus or lysis of infected cells in the presence of complement, some of the antibodies against fragmented HA possess hemagglutination-inhibition activity.

All of the antibody preparations are capable of binding to purified HA1, but only those raised against fragments containing the antigenic polypeptide, HA1CN1, are capable of binding to intact virus. This indicates that within the intact virion the N-terminal CNBr fragment of HA1 carries immunogenic activity.
Antigen-Specific Immunocompetency, B Cell Function, and Regulatory Helper and Suppressor T Cell Activities in Spontaneously Autoimmune Mice
Authors: Creighton W. Dodson , Katz David H. , Dixon Frank J.
Published: December 2022

A comprehensive analysis of the immunocompetency of three murine strains displaying spontaneous autoimmune disease was conducted at various times during the lifespan of these mice. Qualitative and quantitative comparisons have been made for antibody responses to well defined hapten-carrier conjugates between young and old autoimmune as referenced against nonautoimmune inbred mice of the same ages. Responses of intact autoimmune and normal murine strains in two immunoglobulin classes, i.e., IgG and IgE, failed to reveal any general pattern of abnormal immune reactivity that could be ascribed solely to the autoimmune phenotype; grossly deficient antibody-producing capacities that were observed in old mice of the MRL/1 strain do not necessarily relate to the existence of autoreactivity in such animals.

In addition to studies in intact mice, the immunocompetency of isolated B lymphocyte and regulatory T lymphocyte populations of these autoimmune mice were assayed for functional abnormalities by utilizing adoptive transfer systems. No significant differences were observed between young and old autoimmune mice and normal control strains either in the antibody-producing abilities of adoptively primed hapten-specific B lymphocytes or in regulatory functions of antigen-specific helper and suppressor T cells. Even in those circumstances in which intact mice showed marked deficiencies in antibody responses, i.e., old MRL/1, lymphocytes removed from the native environment of such mice displayed normal reactivity patterns in adoptive transfer situations.

Of special note are the results presented herein that clearly demonstrate that antigen-specific regulatory T cell functions of both helper and suppressor types are, in fact, normal in both old as well as young animals of all three autoimmune murine strains. These findings raise serious questions about previous concepts concerning the existence of any single defect or combination of immunoregulatory defects as universal pathogenetic mechanisms in the development of autoimmune diseases.