bluuebunny/crossref_metadata_2025 huggingface dataset:
Inhibiton of IL 2 production after human allogeneic bone marrow transplantation.
Authors: Azogui O , Gluckman E , Fradelizi D
Published: December 2022

Bone marrow transplantation (BMT) is currently used to treat patients with severe aplastic anemia or leukemia. Despite the use of an HLA identical sibling donor, however, the survival after BMT is reduced by the occurrence of two major immunologic complications: graft-vs-host disease and a long-lasting immune deficiency responsible for late infections. This immune deficiency could be explained by an imbalance of lymphocyte subpopulations reconstituted after transplant. The aim of the present work was to study the helper function by measuring the production of interleukin 2 (IL 2). This lymphokine is responsible for the amplification of the effector phase of immunity. A consecutive series of 34 patients was tested for IL 2 production after BMT. This production was absent or low in 32 of 34 patients for at least 2 yr after BMT. The mechanism of this low IL 2 production was investigated. Irradiation of patients' lymphocytes in vitro with low dose gamma-rays partially restored the IL 2 production after 6 mo of evolution. The IL 2 production was not restored or was slightly affected by irradiation early after BMT. These results suggest that the lack of immune reconstitution after BMT may be caused by the lack of IL 2-producing cells and/or the increased activity of suppressor cells of the helper function. This suppression is radiosensitive.
Altered expression of lymphocyte Fc alpha receptor in selective IgA deficiency and IgA nephropathy.
Authors: Adachi M , Yodoi J , Masuda T , Takatsuki K , Uchino H
Published: December 2022

To study the expression of FcR specific for IgA (Fc alpha R) on human peripheral lymphocytes (PBL), PBL from normal donors were incubated with 300 to 500 micrograms/ml MOPC 315 IgA having anti-trinitrophenyl (TNP) antibody activity at 4 degrees C or 37 degrees C for 60 min. Under this condition, less than 2% of total cells could form rosettes with TNP-coated ox red blood cells (TNP-ORBC). When cultured with MOPC 315 IgA at 37 degrees C for 18 hr, however, there was a dose-dependent increase of the rosette-forming cells (RFC) binding TNP-ORBC. Because 15 to 20% of the total cells bound TNP-ORBC but not unsensitized ORBC, the rosette formation appeared to be due to the cytophilic binding of IgA to the cells. The binding of MOPC 315 IgA was competed by TEPC 15 IgA and human myeloma IgA, but not by murine myeloma proteins of other classes, indicating that the receptor is specific for IgA. Fc alpha R was induced on 15 to 20% of fractionated T and B cells, as well as on 15 to 18% of concanavalin A-(Con A) activated lymphocytes when cultured with IgA. The induction of the receptor was dependent on protein and RNA synthesis, but not on DNA synthesis as suggested by the sensitivity to metabolic inhibitors, such as mitomycin C, actinomycin D, puromycin, and cycloheximide. In five patients with selective IgA deficiency (serum IgA, 0 to 4 mg/dl), only 5.1% +/- 1.7 of PBL formed rosettes with TNP-ORBC after culture with MOPC 315 IgA, whereas 12.5% +/- 2.5 of PBL from normal donors (serum IgA, 90 to 330 mg/dl) formed rosettes. Fc alpha R was induced on more than 15% of the cells from these patients, however, when cultured with IgA in the presence of a conditioned medium obtained from mixed lymphocyte culture from two normal donors. The results suggested that the abnormality in the patients' PBL might be in the induction mechanism rather than in the number of precursor cells that could express Fc alpha R in the presence of IgA. On the other hand, Fc alpha R was induced on 10.4% +/- 1.5 of PBL from the patients with IgA nephropathy (serum IgA, 382 +/- 11 mg/dl) when they were incubated with IgA for 1 hr at 37 degrees C. Because Fc alpha R on normal PBL was not induced by 1 hr of incubation with IgA, it appeared that the receptor was already expressed in vivo on the cells of these patients.
The effect of the X-linked immune deficiency gene (xid) upon the Y chromosome-related disease of BXSB mice.
Authors: Smith H R , Chused T M , Steinberg A D
Published: December 2022

BXSB mice develop a lupus-like disease characterized by B cell hyperplasia, hypergammaglobulinemia, autoantibodies, nephritis and coronary artery disease. To determine the subset of B cells responsible for disease in these mice, we bred a congenic BXSB.xid strain (greater than 99.2% inbred) with the xid gene that deletes a subset of splenic B cells. Because BXSB disease is associated with the Y chromosome, BXSB males and the autoimmune cross (NZB X BXSB)F1 males were studied. BXSB.xid males had profoundly reduced lymphoid hyperplasia, hypergammaglobulinemia, autoantibodies, renal disease, cardiac disease, and markedly prolonged survival. (NZB.xid/+ X BXSB)F1 males also demonstrated a marked protection associated with the xid gene. These studies suggest that the autoimmune disease of BXSB males is dependent upon the B cell subset deleted by xid.
A rapid biologic assay for the detection of interleukin 1.
Authors: Conlon P J
Published: December 2022

The detection of human peripheral blood-derived IL 1 was accomplished by a modification of the monokine-dependent conversion assay of the cell line LBRM-33 from the IL 2 nonproducer to producer phenotype after incubation with the monokine and lectin. The conversion of the LBRM-33 subclone, 1A5B6, was modified by directly adding the IL 2-dependent cell line CTLL-2 to metabolically inactivated LBRM-33. This enabled the more rapid identification of macrophage-derived IL 1 and was approximately 1000 to 10,000 times more sensitive than the traditional thymocyte proliferation assay. It is hoped that with the assay described here, it will be feasible to more accurately screen for the presence of this important immunoregulatory molecule in an immune response.
Dissociation in expression of MB1/MT1 and DR1 alloantigens in mutants of a lymphoblastoid cell line.
Authors: DeMars R , Chang C C , Marrari M , Duquesnoy R J , Noreen H , Segall M , Bach F H
Published: December 2022

Cytotoxicity tests with alloantisera were used to study the expression of HLA-D region antigens in HLA-DR-null mutants of a human lymphoblastoid cell line. The initial cell line contained just one copy of the MHC as a haplotype that included DR1 and MB1/MT1. Gamma ray mutagenesis of the single haplotype cells followed by selection with complement and an anti-DR monoclonal antibody were then used to isolate DR-null mutants. Two categories of mutants were identified with a panel of alloantisera. Expressions of DR1 and MB1/MT1 were simultaneously lost in four mutants. Nine mutants still expressed MB1/MT1 but had lost the expression of DR1. The dissociated loss of expression of MB1/MT1 and DR1 antigens is evidence for separate genetic control of these alloantigens. The methods used exemplify a versatile approach for conveniently inducing separations of closely linked loci of the MHC.
Enhancement of murine T cell I-J expression by limited proteolysis.
Authors: Klyczek K K , Hullett D A , Hayes C E
Published: December 2022

I-J-encoded structures on peripheral T cells and thymocytes appear normally to be blocked or shielded by material that is susceptible to proteolysis. Limited proteolysis with trypsin, papain, pronase, or chymotrypsin increased the number of peripheral T cells and thymocytes lysed by anti-I-Jk serum and complement. Proteolysis did not induce I-Jk expression on B cells or on negative strain T cells. Increased lysis was enzyme concentration and time dependent and was not due to increased susceptibility of protease-treated cells to lysis by antibody plus complement; proteolysis rendered T cells and thymocytes less susceptible to lysis by anti-H-2Kk, anti-H-2Dd, and anti-Lyt-2 antibodies. Absorption experiments showed that I-Jk determinant density was increased in the protease-treated T cell population. The I-Jk determinants detected are proteins or glycoproteins; extended proteolysis removed these molecules from the T cell surface. Treatment of T cells or thymocytes with activated macrophage culture supernatant containing proteolytic activity produced a small but reproducible increase in I-Jk expression. Proteolysis of lymphocyte membranes, possibly mediated by macrophages, may have a role in cellular differentiation and immune activation.
Alternative pathway of complement activation by human lymphoblastoid B and T cell lines.
Authors: Praz F , Lesavre P
Published: December 2022

Other investigators have reported activation of the alternative complement (C) pathway in homologous and heterologous serum by a variety of human lymphoblastoid B cell lines. Their ability to activate the C was associated with Epstein Barr virus transformation of the cells. We report that some human lymphoblastoid T cell lines, lacking EBV-DNA in their genome, do activate the human alternative C pathway with no participation of immunoglobulin or specific antibodies. Nevertheless, activation measured by C3 deposition on the cell surface was weaker than with the B cell lines so far studied; in relation to the RAJl cells, C3 deposition on JURKAT, MOLT 4, HSB2, and 1301 was, respectively, 68, 38, 28, and 19%. Furthermore, C3 deposition on JURKAT T cell line requires D, whereas RAJl cells provide a proteolytic activity able to mimic D, and which, unlike D, can be controlled by serum protease inhibitors. Although the ability to activate the AP seems to be a largely shared property of the human lymphoblastoid B and T cell lines, the situation was strikingly different with normal and mitogen-stimulated lymphocytes or with acute leukemic cells, which led to a negligible AP-dependent C3 deposition compared with the level observed with the lymphoblastoid cell lines. Membrane sialic acid content was determined for every cell tested and revealed no relationship with their ability to activate the AP. The two EBV+ B cell lines tested led to a comparable AP activation regardless of the presence of the C3b receptor.
Ocular herpes simplex virus infection is diminished by depletion of B lymphocytes.
Authors: Jordan C , Baron S , Dianzani F , Barber J , Stanton G J
Published: December 2022

Rabbit anti-mouse mu-chain serum was used to deplete specifically IgM-bearing B cells and B cell function in BALB/c mice. B cell-depleted and normal littermates were infected via the cornea with herpes simplex virus type 1 (HSV-1) at 4 to 5 wk of age. B cell-depleted mice had a reduced number of lymphocytes bearing surface immunoglobulin and a greatly reduced ability to produce antibody. Mortality was reduced from 89% in normal controls to 42% in B cell-depleted mice (p less than 0.001) and peak virus shedding from eyes was also reduced as much as 90% in the B cell-depleted mice (p less than or equal to 0.0005). These results support the hypothesis that a B cell function of normal mice somehow contributes directly or indirectly to the enhanced mortality of mice infected with HSV.
Biochemical analysis of a 130,000 molecular weight glycoprotein on human melanoma cells.
Authors: Albino A P , Lloyd K O , Ikeda H , Old L J
Published: December 2022

A monomeric sialoglycoprotein of 130,000 molecular weight (gp 130) is a membrane protein detected by mouse monoclonal antibodies on human melanoma cells and in lesser amounts on a wide range of normal and malignant cell types. Eight monoclonal antibodies reacting with gp 130 detect at least four spatially distinct epitopes on the exposed surface of the gp 130 molecule. Biosynthetic studies have shown that gp 130 is synthesized through two precursor forms: a 100 kD glycosylated species and an 80 kD unglycosylated species, presumably the primary translational product of the encoding mRNA.
Frequency analysis of functional immunoglobulin C and V gene expression in murine B cells at various ages.
Authors: Hooijkaas H , Preesman A A , Van Oudenaren A , Benner R , Haaijman J J
Published: December 2022

The frequencies of lipopolysaccharide- (LPS) reactive B cells and their antibody specificity repertoire have been determined in the spleen and bone marrow (BM) of mice at different ages. A limiting dilution culture system was employed that allows the growth and development of every LPS-reactive B cell into a clone an IgM-secreting cells that are capable of switching to other Ig heavy chain isotypes (C gene expression). The secretion of IgM and IgG1 was assessed in the protein A plaque assay, whereas specific IgM antibody-secreting cells (V gene expression) were detected with the use of plaque assays specific for various heterologous erythrocytes and sheep red blood cells (SRBC) coupled with a number of different haptens. The frequencies of LPS-reactive B cells in the spleen and BM of C3H/Tif, C57BL/Ka, BALB/c, and CBA/Rij mice appeared to be similar in 6- to 12- and 100-wk-old animals, as was the switch frequency to IgG1 secretion in three strains tested. Moreover, no age-related changes were observed in the frequencies of antigen-specific B cells within the pool of LPS-reactive B cells in the spleen and BM of C57BL/Ka mice. The frequencies ranged from 1 in 10 to 1 in 20 for NIP4- and NNP2-SRBC, from 1 in 50 to 1 in 100 for TNP30-SRBC, and from 1 in 1000 to 1 in 4000 for SRBC, HRBC, and GRBC. The specificity repertoire of the "spontaneously" occurring ("background") IgM-secreting cells in the spleen and BM, on the other hand, did differ between young and old C57BL/Ka mice. During aging the frequencies of the tested specificities decreased in the spleen but increased in the BM. Our data indicate that in unintentionally immunized mice the clonal selection of B lineage cells by antigen takes place at the level of the mature, antigen-reactive B cell.
Epstein-Barr virus induces normal B cell responses but defective suppressor T cell responses in patients with systemic lupus erythematosus.
Authors: Tsokos G C , Magrath I T , Balow J E
Published: December 2022

Epstein-Barr virus (EBV) is a polyclonal B cell activator, independent of helper T cells, which induces the generation of suppressor T cells in vivo and in vitro. Given the complexity of the immunologic abnormalities in patients with systemic lupus erythematosus (SLE), we used EBV as a tool to examine the following questions: a). Are SLE B cells primarily defective? and b). Does EBV stimulate the generation of suppressor activity from SLE T cells? It was found that B cells from SLE patients infected with EBV in vitro generate plaque-forming cell (PFC) responses that are similar to those raised by normal B cells infected with EBV within the first 14 days of culture. T cells from SLE patients, in contrast to T cells from normal individuals, cultured with autologous B cells plus EBV fail to develop the expected normal decrement of PFC during the late phase of the in vitro culture (day 14). However, B cells from SLE patients are susceptible to suppression as mixed cultures of SLE B cells and normal allogeneic T cells showed a pattern of PFC response to EBV similar to that of the co-culture of normal B cells with normal T cells. T cells from SLE patients, in analogous mixed cell cultures, failed to suppress either normal B cells or allogeneic SLE B cells. The above experiments indicate that the B cells are not intrinsically defective in SLE patients; rather, a specific T cell abnormality contributes to the lack of normal immunoregulation of certain B cell responses in SLE.
Release of platelet activating factor in rabbits with antibody-mediated injury of the lung: the role of leukocytes and of pulmonary endothelial cells.
Authors: Camussi G , Pawlowski I , Bussolino F , Caldwell P R , Brentjens J , Andres G
Published: December 2022

This paper describes the release of platelet-activating factor (PAF) into the circulation of rabbits with acute pulmonary injury induced by antibody reacting with pulmonary endothelium. Eight rabbits were injected i.v. with 2 mg/kg of body weight of goat anti-rabbit lung angiotensin-converting enzyme gamma-globulin (GtARbACE). All animals developed acute pneumonitis, characterized by severe endothelial damage, accumulation of polymorphonuclear leukocytes (PMN) and platelets (Plt) in the lumina of alveolar capillaries, and deposits of goat IgG and rabbit C3 along alveolar capillary walls. Six of the rabbits died from acute pulmonary edema. PAF was detected in the plasma of all animals within 5 min after injection of GtARbACE. Five other rabbits were depleted of leukocytes by nitrogen mustard and then injected with 2 mg/kg of body weight of GtARbACE. In three of these rabbits release of PAF was demonstrated, though in amounts smaller than in non-leukocyte-depleted rabbits; all three animals died from pulmonary edema. After injection of 0.03 mg/kg of body weight of GtARbACE in six additional rabbits, three of them leukocyte-depleted, small amounts of PAF were detected in the circulation. None of these six rabbits died of pulmonary edema. PAF release was not observed in ten rabbits injected i.v. with 2 or 0.03 mg/kg of body weight of normal goat gamma-globulin. In separate experiments in vitro, incubation of isolated lung or thoracic aorta with GtARbACE resulted in deposits of goat IgG along endothelia and significant release of PAF. PAF was also released from endothelial cells removed from thoracic aorta by cellulose acetate paper and then incubated with GtARbACE. When segments of thoracic aorta were stripped of endothelium and then incubated with GtARbACE, PAF release could not be shown. The data obtained are consistent with the interpretation that PAF released into the circulation after binding of GtARbACE to the endothelia of lung and aorta originates from leukocytes and from lung and thoracic aorta endothelial cells.
Activation of nonspecific killer cells by interleukin 2-containing supernatants.
Authors: Teh H S , Yu M
Published: December 2022

In the absence of specific antigen stimulation, nonspecific killer cells were induced by culturing C57BL/6 lymph node or spleen cells with interleukin 2-containing supernatants. These supernatants were obtained from stimulation of either rat spleen cells with concanavalin A or a variant of the T cell lymphoma, EL4 (H-2b) with phorbol myristic acetate. The ability of the EL4 supernatant to induce nonspecific killer cells was abrogated by absorption with an interleukin 2-dependent T cell line or by concanavalin A-stimulated spleen cell blasts, but not by lipopolysaccharide-stimulated spleen cell blasts or by a non-interleukin 2-producing EL4 line. Partially purified interleukin 2 from EL4 supernatants could also support nonspecific killer cell induction. The induction of cytolytic cells by interleukin 2 is sensitive to gamma-irradiation and has a D omicron of 120 rad. The nonspecific killer cells induced are likely cytotoxic T lymphocytes; the majority of the precursor and effector cells bear the Thy-1 alloantigen marker. These nonspecific killer cells killed a broad spectrum of target cells, including concanavalin A- and lipopolysaccharide-induced splenic blasts of syngeneic or allogeneic mice, a syngeneic tumor, and a cloned allogeneic cytotoxic T cell line. The frequency of precursors for nonspecific killer cells in C57BL/6 lymph node and spleen cells are 1/7000 and 1/12,000, respectively. Clonal analyses revealed that these nonspecific killers exhibit heterogeneity with respect to their target cell specificities. The induction of nonspecific killers by interleukin 2-containing supernatants is partially dependent on nylon wool-adherent cells; in antigen-stimulated cultures the most specific killer cells were obtained from cultures in which nylon wool-nonadherent lymph node responder cells were stimulated with nylon wool-nonadherent allogeneic splenic stimulator cells that were treated with anti-Thy-1 antibody and complement. The relevance of these findings with respect to the frequencies and fine specificities of cytotoxic T lymphocytes generated in interleukin 2-supplemented cultures is discussed.
Role of interferon in human natural killer activity against target cells infected with HSV-1.
Authors: Bishop G A , Glorioso J C , Schwartz S A
Published: December 2022

Human natural killer (NK) cells show high cytotoxic activity against target cells infected with herpes simplex virus type 1 (HSV-1). Substantial amounts of interferon (IFN) were generated in co-cultures of NK effector cells and infected target cells; however, the cytotoxic activity seen against a specific infected cell target did not correlate with the amount of IFN induced. The production of IFN increased steadily from 4 to 18 hr of co-culture, as did NK activity; however, IFN production peaked 4 hr later than NK activity. Pretreatment of NK effector cells with exogenous IFN increased cytotoxic activity against all targets tested, but the differential pattern of reactivity against cells infected with wild type and mutant viruses was unaltered. When effector cells were treated with the RNA synthesis inhibitor actinomycin D before co-culture with virus-infected targets, IFN production was markedly reduced, without a concomitant reduction in cytotoxicity. Similarly, the addition of anti-IFN antiserum to co-cultures greatly decreased the available IFN present, but had no effect on NK activity. We conclude that the induction of cytotoxic activity in co-cultures of NK effector cells and HSV-1-infected target cells is independent of the induction of IFN.
Functional characteristics of histamine receptor-bearing mononuclear cells. I. Selective production of lymphocyte chemoattractant lymphokines with histamine used as a ligand.
Authors: Center D M , Cruikshank W W , Berman J S , Beer D J
Published: December 2022

Mitogens and antigens have been the traditional ligands for activating lymphocytes in vitro for the elaboration of lymphokines. Recently, histamine, by interaction with histamine-type 2 receptors on T lymphocytes, has been found to induce the production of one lymphokine, histamine-induced suppressor factor (HSF), that inhibits lymphocyte proliferation and lymphokine production in vitro. Because the biologic effects of HSF appear to be confined to alterations in lymphocyte function, we assessed the ability of soluble products of histamine-stimulated human blood mononuclear cells to affect another lymphocyte function, motility. Utilizing a modified Boyden chamber assay to assess lymphocyte migration, we identified chemoattractant activity for human blood and rat splenic T lymphocytes in histamine-induced mononuclear cell supernatants. No neutrophil or monocyte chemoattractant activity was present. Sephadex G-100 gel filtration of histamine-induced supernatants showed the lymphotactic activity eluted with a 56,000 m.w. This activity was cationic as determined by its elution pattern from a Sephadex QAE anion exchange matrix with a single pl of 9.0 to 9.4 determined by isoelectric focusing in sucrose. Its biologic activity is predominantly chemokinetic in nature, is stable to heating at 56 degrees C for 30 min, but is sensitive to the effects of trypsin and neuraminidase. These physicochemical and functional characteristics establish it as identical to a recently described concanavalin A-induced (Con A) lymphotactic lymphokine (LCF). Mononuclear cells that did not adhere to a histamine affinity matrix were unable to produce LCF when subsequently stimulated with histamine or Con A. Mononuclear cells incubated with histamine and diphenhydramine produced LCF; the addition of cimetidine eliminated LCF production. In fact, supernatants from cells incubated with histamine and cimetidine significantly inhibited lymphocyte migration, a phenomenon explainable by the two regions of lymphocyte migration inhibitory activity that were present in the Sephadex G-100 chromatography of crude histamine-induced supernatants. These data suggest that a subset of lymphocytes defined by the presence of histamine-type 2 receptors is capable of producing LCF while cells that bear histamine-type 1 receptors produce lymphocyte migration inhibitory activity.
Human granulocyte surface molecules identified by murine monoclonal antibodies.
Authors: Skubitz K M , Pessano S , Bottero L , Ferrero D , Rovera G , August J T
Published: December 2022

We have investigated the nature of the antigens recognized by four classes of mouse anti-human monoclonal antibodies that characteristically reacted with neutrophilic granulocytes and their precursor cells, but not with monocytes or other normal hemopoietic cells. The antigenic targets of the majority (9/12) of the independently isolated monoclonal antibodies were present on two surface glycoproteins (Mr 145,000 and 105,000) and glycolipids. This antigen(s) was also detected on granulocyte precursor cells, including the bone marrow granulocyte/monocyte progenitor cells (CFU-GM). The same antigen(s) detected by these monoclonal antibodies was also present in non-hemopoietic cell lines (colon carcinoma and neuroblastoma). Three other antigens, defined by monoclonal antibodies AHN-8, L12.2, and L13.1 and present on granulocytes and their mid-late precursor cells, could not be identified as proteins but were detected in a protein-free glycolipid extract of these cells. The diversity of the antigens was confirmed by cross-competition experiments and by the identification of their different patterns of reactivity with cell lines and bone marrow cells.
Ligand-induced association of rat lymphocyte membrane proteins with the detergent-insoluble lymphocyte cytoskeletal matrix.
Authors: Woda B A , McFadden M L
Published: December 2022

There are two classes of membrane protein capping on the basis of ligand requirements. Surface immunoglobulin (Slg), the prototype of the first class, requires a single ligand for cap induction. RT1 (rat histocompatibility proteins) requires two antibodies for cap induction. The lateral mobility of Slg is relatively restricted compared with RT1. These differences may be due to differential interaction with the cytoskeleton. After ligand binding 71% of Slg becomes detergent insoluble and is associated with the lymphocyte cytoskeletal matrix. The insolubilization occurs at 4 degrees C and is not inhibited by sodium azide or cytoskeleton-active drugs. The insolubilized ligand-receptor complex can be solubilized by a cytoskeleton destabilizing buffer. In contrast, only 20% of RT1 becomes associated with the lymphocytic cytoskeleton after ligand binding. The ligand-induced receptor-cytoskeleton interaction influences capping behavior and may play a role in cell activation.
Sequential induction of phospholipid methylation and serine esterase activation in a B cell differentiation factor (BCDF)-stimulated human B cell line.
Authors: Kishi H , Miki Y , Kikutani H , Yamamura Y , Kishimoto T
Published: December 2022

The incubation of CESS cells with BCDF induced phospholipid methylation, i.e., 3H-methyl incorporation into phosphatidylcholine and phosphatidyl-N-monomethyl-ethanolamine, within 15 to 30 min. A threefold increase of 3H-DFP-binding was also observed in BCDF-stimulated CESS cells 2 hr after stimulation, showing the activation of serine esterase after phospholipid methylation. The inhibitors of methyltransferase, SIBA, adenosine, or HEH showed a dose-dependent inhibitory effect not only on phospholipid methylation but also on serine esterase activation and on BCDF-induced IgG production; DFP inhibited IgG production but not phospholipid methylation. The treatment of cells with DFP in the presence of SIBA abrogated the irreversible inhibitory effect of DFP on IgG production. These results showed that BCDF stimulation induced phospholipid methylation that set the stage for serine esterase activation and IgG induction.
Characterization of alloimmunization-induced T lymphocytes reactive against AKR leukemia in vitro and correlation with graft-vs-leukemia activity in vivo.
Authors: Truitt R L , Shih C Y , Lefever A V , Tempelis L D , Andreani M , Bortin M M
Published: December 2022

We have reported that immunization of H-2k mice with lymphoid cells from various allogeneic strains induced a population of cells that could eliminate first-passage spontaneous AKR leukemia from the spleens of immuno-suppressed AKR (H-2k) hosts. In the present study, we examined the nature of the cells responsible for this graft-vs-leukemia (GVL) reaction and compared them to cytolytic cells detected in vitro. Spleen cells from alloimmunized CBA/J (H-2k) mice were selectively depleted of various subpopulations by treatment with antibody and complement (C), then tested in vivo for GVL reactivity. Cell suspensions depleted of Thy-1.2+, Lyt-1+, or Lyt-2+ lymphocytes had no significant GVL reactivity, whereas suspensions depleted of NK-1.2+ cells retained GVL reactivity. The GVL-reactive cells persisted in H-2-compatible donor mice for up to 56 days. Lyt-1+2+ lymphocytes that were cytotoxic for cultured AKR leukemia cells in vitro could be detected in the spleens of alloimmunized H-2-compatible mice after expansion of the cells in T cell growth factor. Using quantitative limiting dilution cytotoxicity assays, we found that the frequency of leukemia-reactive cytotoxic lymphocytes (CL) in the spleen showed a direct correlation with the GVL efficacy of the cells in vivo. Alloimmunization was essential for induction of the GVL-reactive cell population. CL in alloimmunized mice consisted of heterogeneous cytotoxic specificities; i.e., some CL were leukemia-specific, others lysed only nonleukemic AKR target cells, and a third group mediated killing of both leukemic and nonleukemic target cells. The CL appeared to be H-2 restricted and specific for non-H-2 antigens shared by the AKR leukemia and the alloimmunizing cells.
Antigen-induced lymphomagenesis: identification of a murine B cell lymphoma with known antigen specificity.
Authors: Arnold L W , LoCascio N J , Lutz P M , Pennell C A , Klapper D , Haughton G
Published: December 2022

CH12, a murine B cell lymphoma derived in B10 H-2aH-4bp/Wts mice after transfer of SRBC hyperimmunized spleen cells into an adult-thymectomized, sublethally irradiated, syngeneic recipient, is demonstrated to bear surface IgM specific for a determinant found on SRBC and ChRBC. The Ig specificity has been demonstrated by rosetting assays and complement-dependent hemolysis. The removal of CH12 surface IgM by capping with anti-mu or with anti-CH12 idiotype, but not with anti-gamma or with irrelevant anti-idiotype, eliminated the formation of rosettes between CH12 and SRBC or ChRBC. The absorption of CH12 Ig produced in vitro, with either SRBC or ChRBC but not with HRBC, removed all hemolysin activity against SRBC, demonstrating that only one CH12 product was responsible for the reactivity with both SRBC and ChRBC. CH12 has a surface phenotype of a relatively mature B cell expressing surface Ig (IgM-mu,kappa) and la antigens, but lacking Thy-1 or detectable Fc or C3 receptors. CH12 also expresses the antigen Lyt-1. Growth of CH12 in vivo or in vitro results in the generation of up to 3% direct PFC and serum hemolysin, which shows that CH12 is not irretrievably "frozen". The generation of PFC and serum hemolysin is associated with increased population density, and the rate of PFC and serum hemolysin accumulation cannot be explained by simple cell division. A continuously secreting hybridoma derived from CH12 was used to purify the CH12 IgM to facilitate studies of protein sequence and idiotype.
Anti-mu induces lymphoma in germfree congenitally athymic (nude) but not in heterozygous (nu/+) mice.
Authors: Gershwin M E , Ohsugi Y , Castles J J , Ikeda R M , Ruebner B
Published: December 2022

To further our understanding of the role of host immunity in the development of lymphoid neoplasia, groups of germfree BALB/c nude and nu/+ mice were either followed unmanipulated or were treated, beginning at birth, with anti-mu, normal goat IgG or with lipopolysaccharide (LPS). The survival and development of neoplasia of all groups of animals were monitored up to 2 yr. Nude mice, under germfree and specific pathogen-free (spf) conditions, had a higher incidence of lymphoid neoplasia and reduced survival when compared to nu/+ littermates. The incidence of lymphoid tumors in nude mice under spf or germfree conditions was 7.2 and 8.7%, respectively, in comparison to 0% in nu/+ animals. Treatment of germfree nude mice with anti-mu, but not with goat IgG, increased the incidence of lymphoid tumors to 39%. Anti-mu did not significantly change the incidence of lymphoid neoplasia in nu/+ animals. Treatment of nu/nu and nu/+ mice with LPS, however, led to a several-fold increase in the appearance of neoplasia, to values of 25.4% in nude and 10% in nu/+ mice. Lymphoid neoplasia found in either unmanipulated, anti-mu, or IgG-treated germfree or spf mice included Thy-1.2+, surface IgM+, and IgG+ tumors. In contrast, all the lymphomas found in LPS-treated mice were surface IgM+. Thus, whereas LPS may have generated a relatively homogeneous group of tumors, anti-mu may have randomly increased the normal incidence of spontaneous tumors. Moreover, although there was significant variation in the histologic appearance of tumors, both within treatment groups as well as in different areas of the same animal, only LPS-treated mice were regularly noted to have distant nonlymphoid involvement, with lesions found in liver, lung, and kidney. In contrast, the incidence of nonlymphoid neoplasia was similar and was less than 2.5% in all groups.
Tumor-associated macrophages of mouse mammary tumors. II. Differential distribution of macrophages from metastatic and nonmetastatic tumors.
Authors: Mahoney K H , Fulton A M , Heppner G H
Published: December 2022

Macrophage subpopulations were isolated by counterflow centrifugal elutriation from a number of closely related mammary tumors originally derived from a single spontaneous tumor in a BALB/cfC3H mouse. Each tumor line had its own distinct characteristics with regard to tumor cell distribution, macrophage size (density), macrophage content, and macrophage distribution. Although there is no correlation with the metastatic potential of the tumors for the first three characteristics listed, distinct differences are apparent between metastatic and nonmetastatic tumors in macrophage distribution into the elutriated fractions. The distribution of the tumor-associated macrophages was shifted toward the later fractions in the metastatic tumors (lines 66 and 410.4), with approximately 40% more macrophages in fractions 3 and 4 than in those fractions from the nonmetastatic tumors (lines 67 and 168). The levels of three ectoenzymes, leucine aminopeptidase (LAP), alkaline phosphodiesterase I (APD), and 5' nucleotidase (5'N) were determined for macrophages from the elutriated fractions as well as for the total tumor digest. No significant differences were observed in absolute levels of either LAP or APD between most of the fractions from the different tumor lines. Differences were seen between the fractions in levels of 5'N among the lines and in ADP levels between lines 66 and 168 from the total tumor digest. Striking differences in ectoenzyme levels were apparent when the activity was normalized to macrophage surface area: macrophages from line 66 tumors were 3- to 17-fold higher in their LAP concentrations than were macrophages from either 67 or 168 tumors. Each tumor line had a unique and reproducible pattern of macrophage-associated prostaglandin E (PGE) synthesis. The three nonmetastatic tumor lines showed peak PGE levels in fraction three with lower levels in fraction 2, whereas the highly metastatic tumor yielded more active macrophages in fraction 2 vs fraction 3. Changes in PGE levels were associated with differing ectoenzyme levels. Macrophages in fraction 3 from metastatic tumors were enriched for populations with high LAP activity and low PGE levels. Conversely, macrophages from nonmetastatic tumors exhibited lower LAP activity in fraction 3 vs 2 and increasing PGE levels. Thus, differences between metastatic and nonmetastatic mammary tumors were seen in 1) distribution of macrophages in the elutriation fractions, 2) size of infiltrating macrophages, 3) macrophage ectoenzyme concentrations, and 4) macrophage-associated PGE levels.
Prevention of lethal, minor-determinate graft-host disease in mice by the in vivo administration of anti-asialo GM1.
Authors: Charley M R , Mikhael A , Bennett M , Gilliam J N , Sontheimer R D
Published: December 2022

Graft-vs-Host disease (GVHD) remains a devastating problem in human bone marrow transplantation (1, 2). Because removal of Thy-bearing cells from the donor inoculum has prevented GVHD in murine models (3, 4), it has been hoped that a similar cell surface antigen or combination of antigens could be found in humans. Unfortunately, treatment of human donor cells with various T cell antisera has not yet been successful in preventing GVHD (5). Encouraging results have been reported in five patients who received bone marrow depleted of T cells by the sequential use of soybean agglutinin and the differential sedimentation of cells forming rosettes with sheep red blood cells (6). Although donor T cells are thought to be necessary for initiating GVHD, the immunopathogenesis of GVHD is still not understood. Because donors and recipients are routinely major histocompatibility complex matched and chosen to be nonreactive in mixed lymphocyte cultures human GVHD is thought to result from minor histocompatibility antigen disparities. Lopez and coworkers (7, 8) found a strong association between the incidence of human GVHD and the pretransplant levels of natural killer (NK) activity of the recipients; when the recipient NK activity was low, GVHD rarely developed. They speculated that the NK cell lineage is serving as an important stimulator-inducer. We therefore examined the in vivo effects of anti-asialo GM1 on a murine model of GVHD based on minor antigen disparity. This antiserum has several immunologic effects, including a profound NK suppression. We found that the mice treated with this antibody have normal survival rates, even though they do develop histologic GVHD in the skin. This finding suggests the possibility of a new prophylactic approach to human GVHD and raises many questions regarding the function of asialo GM1-bearing cells in immune regulation.
Role of Ly-1:Qa1- and Ly-1:Qa1+ inducer T cells in activation of Ly-23 effectors of suppression of antibody production in mice.
Authors: Eardley D D , Hu S K , Gershon R K
Published: December 2022

Ly-2+ effectors of T cell-mediated suppression require inducing signals from antigen and a helper cell bearing the Ly-1+:Qa1+ surface phenotype. In this report, we have further examined the helper cell requirements for suppressor cell induction of antibody production in mice. By using the T cell subset education procedure in vitro, we have activated T cells to sheep red blood cells (SRBC) antigens and then purified Ly-2 cells before testing for suppressor activity in assay cultures of defined T and B cell subsets. We have confirmed our previous observations that Ly-1+:Qa1+ cells are required for activation of T suppressors, but have found that under the appropriate conditions, there is not a strict requirement for the Ly-123 subset of T cells. Furthermore, if Ly-23 cells are stimulated in the presence of Ly-1+:Qa1- T cells, effective suppressors can be obtained only if a source of Ly-1:Qa1+ inducers is added to the assay culture. If Ly-23 cells are activated by antigen in the absence of Ly-1 cells, subsequent exposure to the Ly-1+:Qa1+ subset under the conditions tested here is not sufficient to activate suppressors. These results show that effectors of suppression, like B cells and cytotoxic T lymphocytes, may respond to two helper cells.
Clonal analysis of T cell unresponsiveness to alloantigens induced by neonatal injection of F1 spleen cells into parental mice.
Authors: Feng H M , Glasebrook A L , Engers H D , Louis J A
Published: December 2022

Alloantigen-specific T cell precursors present in spleen cell populations obtained from parental mice that were rendered tolerant by the neonatal injection of F1 spleen cells were quantitated by clonal analysis, i.e., limiting dilution techniques. The results demonstrate that the frequencies for cytolytic as well as TCGF-producing (helper) and MAF-producing T cell precursors were dramatically reduced in tolerant spleen cell populations. The unresponsive state was antigen specific and could not be transferred in vitro in cell mixing experiments. Therefore, clonal deletion (or inactivation) would appear to represent a major mechanism for the observed tolerant state in this model system.
Activation of type III pneumococcal polysaccharide-specific suppressor T cells in cyclophosphamide-treated mice requirement for recognition of antigen and I-J determinants on antigen coupled to syngeneic spleen cells.
Authors: Braley-Mullen H
Published: December 2022

Type III pneumococcal polysaccharide (S3) coupled to spleen cells (S3-SC) is a potent tolerogen in mice. Although tolerance induced by S3-SC has been shown to be T cell dependent, T cells from tolerant mice are unable to actively suppress the antibody response to S3 when transferred to normal mice. However, when mice are given cyclophosphamide (Cy) before injection of S3-SC, suppressor T cells (Ts) are induced and can be detected by their ability to suppress the response to S3 after transfer to normal mice. The Ts induced by S3-SC in Cy-treated mice are antigen specific and can bind to and be eluted from plates coated with a soluble membrane preparation of S3-SC (S3-SCSM) prepared from mice that are syngeneic with mice in which Ts are induced. Allogeneic S3-SCSM, free S3, or S3 coupled to non-membrane carriers do not bind the Ts. Moreover, under the same conditions, detectable Ts were not induced by free S3 in either normal or Cy-treated mice. These results suggest that these Ts are activated by, and have receptors for, S3 plus a self cell membrane determinant. The self cell membrane determinant required for Ts induction is apparently I-J since Ts are induced only when the antigen-coupled spleen cells share I-J with the Cy-treated mice in which Ts are activated. The necessity to treat mice with Cy to obtain detectable Ts is presumably due to the fact that S3-SC activates a contrasuppressor T cell in normal mice which interferes with Ts function, and this cell is eliminated by Cy.
Phenotype of the accessory cell necessary for mitogen-stimulated T and B cell responses in human peripheral blood: delineation by its sensitivity to the lysosomotropic agent, L-leucine methyl ester.
Authors: Thiele D L , Kurosaka M , Lipsky P E
Published: December 2022

The lysosomotropic compound L-leucine methyl ester (Leu-OMe) was utilized to delineate the phenotype of the accessory cells involved in human B and T cell activation in vitro. Leu-OMe was shown to cause lysosomal disruption and selective death of human monocytes (M phi). After 30-45 minute incubations with this agent, human peripheral blood mononuclear cells (PBM) were nearly completely depleted of M phi. Associated with this M phi depletion, PBM were rendered unresponsive to a variety of T and B cell mitogens including the plant lectins phytohemagglutinin, concanavalin A, and pokeweed mitogen as well as the oxidative mitogens sodium periodate and neuraminidase plus galactose oxidase. Leu-OMe mediated loss of responsiveness was the result of a selective loss of an accessory cell necessary for each of these responses since reconstitution was accomplished by the addition of a M phi-enriched adherent cell population. While intact adherent cells could reconstitute responsiveness, crude M phi supernatants or highly purified human IL 1 alone were ineffective. Further identification of the Leu-OMe sensitive accessory cell indicated that it was entirely contained within the fraction of the adherent population identified by the monoclonal anti-M phi antibody, 63D3. The mechanism by which Leu-OMe Killed M phi was dependent on the lysosomal content of these cells, since agents that altered lysosomal enzyme activity such as chloroquine or NH4Cl protected M phi from Leu-OMe. Thus, the selective killing of M phi by Leu-OMe appeared to relate to the characteristically rich endowment of lysosomes within these cells. These results support the conclusion that a lysosome-rich, leucine methyl ester-sensitive, intact M phi identified by the monoclonal anti-M phi antibody 63D3 is the circulating accessory cell required for mitogen-triggered human B and T cell activation.
Permissive role of calcium in the inhibition of T cell mitogenesis by calmodulin antagonists.
Authors: Cheung R K , Grinstein S , Gelfand E W
Published: December 2022

The importance of Ca++ in the initiation of lymphocyte activation and mitogenesis has been supported by several studies. Because calmodulin functions as the intracellular mediator of the effects of Ca++, it likely plays a major role in the regulation of lymphocyte function. We have examined the effects of known calmodulin antagonists, the phenothiazines, on lectin-induced T cell mitogenesis and have shown a central role for Ca++ uptake in the expression of a phenothiazine-sensitive stage after lectin activation. The drug effects were observed only if the cells were previously activated by PHA or the ionophore A23187, and only in the presence of Ca++. These effects were restricted to a defined time period (5 hr) after lectin activation. The data support the concept that calmodulin is the target for the phenothiazine effects and demonstrate the permissive role of Ca++ in the mediation of these events.
Alloantigen-specific cytotoxic and suppressor T lymphocytes are derived from phenotypically distinct precursors.
Authors: Damle N K , Mohagheghpour N , Hansen J A , Engleman E G
Published: December 2022

Although Leu-2+ (OKT8+) T cells activated in the mixed lymphocyte reaction (MLR) mediate both alloantigen-specific cytotoxicity and suppression of alloantigen-induced proliferation, it is not known whether these functions derive from a single cell type or phenotypically distinct cells. This study was undertaken to examine the alloantigen-specific cytolytic and suppressor potential of two subpopulations of Leu-2+ cells distinguishable from one another on the basis of their binding to the monoclonal antibody 9.3. Leu-2+, 9.3+ and Leu-2+, 9.3- populations were purified from peripheral blood, cultured for 7 days with autologous helper/inducer (Leu-3+) cells and allogeneic non-T cells, and reisolated before testing for cytotoxicity and suppression. All detectable alloantigen-specific cytolytic activity was confined to the Leu-2+, 9.3+ subpopulation. Killing by this subset was specific for the HLA-A and B (class I) major histocompatibility complex (MHC) antigens of the priming cell. By contrast, suppression of proliferation was mediated predominantly by the Leu-2+, 9.3- cells, and suppression by this subpopulation was specific for the HLA-DR (class II) MHC antigens of the priming cell. The development of suppression by Leu-2+, 9.3- cells was unaffected by cyclosporin A (CsA), an agent shown previously to block the development of cytolytic but not suppressor cells in MLR. Alloactivated Leu-2+, 9.3+ cells were slightly inhibitory of fresh MLR, but this effect as well as the development of cytolytic cells was completely abrogated by CsA. These results indicate that suppressor and cytolytic Leu-2+ T cells activated in MLR are derived from distinct precursors separable by antibody 9.3.