bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunologic control of the ascites form of murine adenocarcinoma 755. V. Antibody-directed macrophages mediate tumor cell destruction.
Authors: Langlois A J , Matthews T , Roloson G J , Thiel H J , Collins J J , Bolognesi D P
Published: December 2022

An in vitro assay has been developed that mimics the potent in vivo protective capacity of B6 anti-AD755a serum in a passive therapy protocol. In the presence of small volumes of hyperimmune serum or IgG2a antibody, thioglycollate-elicited B6 mouse peritoneal cells (PEC) inhibit the growth of AD755 Cl.10 target cells and other cells provided that they express FLV-related antigens. When hyperimmune serum is replaced by normal serum, or elicited PEC from BALB/c mice are used, growth inhibition of the AD755 Cl.10 target cells is not seen. The latter is in accord with the inability to protect BALB/c mice with the anti-AD755a serum. Tumor cells to which antibody has been pre-bound are not inhibited by thioglycollate-elicited B6 PEC, analogous to the undiminished tumorigenicity of antibody-bound AD755a cells in vivo. These results, along with those presented in the accompanying papers, have led to the construction of a model to explain the specificity and mechanism of the potent in vivo passive protection characteristic of this system. The major features of this model are that the protective IgG2a antibody must first interact with an appropriate host effector cell, presumably of the monocyte/macrophage lineage, and that such an antibody-directed cell then binds to the tumor cell and effects its elimination, possibly by a phagocytic mechanism.
The failure of nonresponder mice to develop IgG memory assessed by in vitro culture with an antigen-LPS conjugate.
Authors: Furman A , Sercarz E E
Published: December 2022

A hen egg-white lysozyme-lipopolysaccharide complex (HEL-LPS) can stimulate an in vitro IgG response, but only from HEL-primed B lymphocytes; unprimed cells only produce an IgM response. These conditions were used to determine whether IgG memory B cells are cryptically induced in B10 nonresponder (H-2b) mice after an HEL injection protocol. The usual i.p. immunization that triggers IgG memory production in congenic responder strain mice fails to yield IgG in vitro from HEL-primed B10 spleen cells after stimulation with HEL-LPS. However, injection protocols immunogenic for B10 mice do engender IgG-memory cells. These results imply that the T helper cell population necessary for triggering B cells to the IgG memory stage cannot develop in the nonresponder mouse, presumably due to HEL-specific T suppressor cells.
Macrophage receptors for IgE: binding of IgE to specific IgE Fc receptors on a human macrophage cell line, U937.
Authors: Anderson C L , Spiegelberg H L
Published: December 2022

Human IgE was found to bind to the human macrophage line U937. Binding was specific for IgE in that it was inhibited by 3 different human IgE myeloma proteins and by Fc fragments prepared from IgE, but not by IgE Fab fragments or by myeloma proteins representative of the 4 IgG subclasses or by a rat IgE myeloma protein. Dissociation and association rates were rapid, with estimated t1/2 of less than 1 min. The Ka was 4.15 X 10(7) liter/mol. Between 53,000 and 93,000 sites per cell were calculated. The U937 receptor for IgE was trypsin sensitive, whereas the receptor for IgG on the same cell was relatively trypsin resistant.
Early dominance of irradiated host cells in the responder profiles of thymocytes from P leads to F1 radiation chimeras.
Authors: Korngold R , Bennink J R , Doherty P C
Published: December 2022

The number of cells in the thymus of [k leads to (b X k)F1] radiation (100 rad) chimeras increases approximately 10-fold between 7 and 14 days after reconstitution with bone marrow. At least 50% of the cells in thymus on day 14 are of host origin and respond to virus presented in the context of both H-2k and H-2b when primed in irradiated, virus-infected (b X k)F1 recipients. Strong CTL responses can be generated from thymocytes of donor origin on day 21. All evidence of a significant host thymocyte component has disappeared by day 28. The responsiveness of 14-day thymocytes is not abrogated by pretreatment of the mice used to make the chimeras with anti-thymocyte serum or by using doses of irradiation as high as 1200 rads to eliminate host components.
Immune memory to a nonmitogenic, thymic independent antigen in mice: variation among inbred strains and possible relationship to oncogenesis.
Authors: Schott C F , Lizzio E F , Inman J K , Merchant B
Published: December 2022

Immune memory to DNP-Ficoll, a nonmitogenic, thymic-independent (TI-2) antigen, was demonstrated in several inbred strains of mice. Direct and indirect splenic plaque-forming cell responses were measured in mice given a secondary challenge with DNP-Ficoll and in appropriate control mice. Strong IgM memory, but no IgG memory, was observed in SJL/J, AKR/J, and C58/J mice. C57L/J mice gave both a strong IgM memory response and a relatively strong IgG memory response to DNP-Ficoll. C57BL/6N, C57BR/cdJ, and A/HeJ mice were unable to mount significant IgM or IgG memory responses to this antigen under an identical immunization schedule. These results are indicative of marked genetic variation in mice in the capacity for B memory cell expression. They also identify a provocative but unexplained association between positive IgM memory responses to DNP-Ficoll in SJL/J, AKR/J, and C58/J mice and a propensity to develop lymphoreticular neoplasia. This association is observed when there is no clear evidence for IgG memory.
T lymphocytes of young and aged rats. I. Distribution, density, and capping of T antigens.
Authors: Gilman S C , Woda B A , Feldman J D
Published: December 2022

The distribution, density, and capping of the T cell antigens W3/13, W3/25, and Thy-1 were studied in lymphocytes of young (3 to 4 mo) and aged (greater than or equal to 27 mo) Brown Norway (BN) rats. The total number of W3/13, W3/25, and Thy-1 positive cells and the percentage of Thy-1 positive cells are reduced in the spleens and lymph nodes of aged rats. Analysis of spleen and lymph node cells in a fluorescence-activated cell sorter indicated that with ageing there is a loss of moderately and brightly stained W3/13 and W3/25 positive cells. The density of W3/13, W3/25, and Thy-1 is reduced on spleen and lymph node cells of old rats. The rate of capping of all 3 molecules is diminished on "old" cells. Colchicine treatment of young cells enhanced capping of all 3 molecules and allowed capping of W3/13 by a single ligand. However, capping on "old" lymphocytes was not affected by colchicine treatment. Cytochalasin D inhibited capping to the same extent on young and "old" cells. These results suggest that the membrane composition and cytoskeleton are altered in T lymphocytes of aged rats.
The murine B cell repertoire responsive to an influenza-infected syngeneic cell line.
Authors: Wylie D E , Klinman N R
Published: December 2022

The conditions for eliciting monoclonal B cell responses to an influenza-infected syngeneic tumor cell line were defined. Although the infected tumor cell stimulated vigorous responses, few clones recognized tumor antigens, whereas over 90% of the monoclonal antibodies recognized viral antigens expressed on the surface of infected tumor cells. Approximately one-third of the virus-specific antibodies reacted with the viral hemagglutinin (HA) or neuraminidase, and the remainder recognized antigens found on the surface of infected cells but not on the virion or uninfected cells. The majority of the antibodies reactive with the virion recognized HA determinants, whereas few reacted with the neuraminidase. Subsequent analysis revealed that most neuraminidase determinants exposed on the intact virion were not available on the surface of infected cells. In contrast, the majority of the HA determinants expressed on the virion were also available on the surface membranes of infected cells, and a highly diverse set of antibodies recognized the HA expressed either on the virion or on the infected cell surface. Finally, differences were noted in the heavy chain isotype of antibodies produced in response to infected cells vs purified virus. A majority of monoclonal responses to purified influenza virus included IgA antibodies, whereas responses to infected cells showed less IgA and a concomitant increase in clones expressing IgG.
Lysis of human solid tumors by autologous cells sensitized in vitro to alloantigens.
Authors: Strausser J L , Mazumder A , Grimm E A , Lotze M T , Rosenberg S A
Published: December 2022

Human lymphocytes sensitized in vitro to allogeneic single-donor cells as well as to pools of allogeneic donor cells were evaluated for their cytotoxic potential against autologous fresh solid tumor cells, autologous tissue-cultured tumor and fibroblasts, fresh peripheral blood lymphoid cells (PBL), and Con A blasts of PBL. Using 4-hr 51Cr release assays, pool-sensitized effector cells lysed autologous tissue-cultured tumor and fibroblast targets in 4 of 4 experiments. Similarly, in 2 of 2 experiments with single-donor allosensitized effectors, autologous tissue-cultured tumor was lysed. No lysis of fresh PBL was seen. Significant lysis of fresh autologous tumor target cells was seen in 10 of 12 experiments with pool-sensitized effectors, though, again, no lysis of autologous PBL was observed. No lysis of autologous Con A blasts was seen in 2 of 2 experiments. Fresh tumor cells, PBL, and Con A blasts appeared equally lysable, since they were equally lysed by effector cells specifically sensitized to alloantigens present on these cells. Cold target inhibition studies demonstrated that cultured tumor and fibroblasts but not PBL shared the determinants recognized by pool-sensitized cells.
Alternative complement pathway-dependent ingestion of fluolite particles by human granulocytes.
Authors: Arnaout M A , Luscinskas F W , Lionetti F J , Alper C A , Valeri C R
Published: December 2022

Fluorescent particles (Fluolite) with an average size of 0.1 micrometers were ingested by human granulocytes after incubation in fresh normal human serum (NHS). Ingestion was assessed by visual counting in a fluorescent microscope of cells containing particles. Ingestion required fresh normal serum and did not occur when serum was heated for 30 min at 50 degrees C or in the presence of ethylenediaminetetraacetic acid (EDTA). It did not occur in serum genetically deficient in C3b inactivator or in C3. Phagocytic activity was restored to C3-deficient serum by purified human C3 and to heat inactivated serum by purified factor B. Opsonic activity was present in NHS containing 5 mM Mg++ and 10 mM ethyleneglycoltetraacetic acid (EGTA) and in human serum genetically deficient in human C components C2 and C5. Agammaglobulinemic sera had normal opsonic activity. Opsonization of particles in this system is mediated through the alternative pathway of C activation, and its measurement serves as a simple quantitative functional assay for this system.
The molecular mechanism of cryoprecipitation and cold agglutination of an IgM lambda Waldenström macroglobulin with anti-Gd specificity: sedimentation analysis and localization of interacting sites.
Authors: Weber R J , Clem L W
Published: December 2022

The physicochemical properties and immunologic specificity of an IgM lambda cryoglobulin with cold agglutinin activity (designated MAT) were investigated. Discrete binding sites involved in cryoprecipitation were shown to exist on the reductive 7S subunits and tryptic Fab mu and Fc mu 5 fragments derived from MAT. Analytical ultracentrifugation indicated the nonprecipitating MAT subunits self-associated into rapidly sedimenting paucidisperse boundaries of 32S, 25S, and 8.7S at 13 degrees C, which were converted to a monodisperse 5.7S boundary at 35 degrees C. Furthermore, MAT Fab mu and Fc mu 5 fragments exhibited association with each other at 13 degrees C but not at 35 degrees C when studied by analytical ultracentrifugation. The cold agglutinin activity of MAT can be inhibited by sialyllactose or treatment of erythrocytes with neuraminidase but not papain. Therefore, the membrane determinants of human erythrocytes are Gd (glycolipid-dependent or protease-resistant glycoproteins) with terminal sialic acid. Treatment of MAT with neuraminidase abolished its precipitating properties, but had no effect on its ability to agglutinate untreated erythrocytes. The cryoprecipitation and cold agglutination phenomena exhibited by MAT therefore seem to involve identical or cross-reacting carbohydrate antigens with terminal sialic acid residues.
Modulation of granulomatous hypersensitivity. II. Participation of Ly 1+ and Ly 2+ T lymphocytes in the suppression of granuloma formation and lymphokine production in Schistosoma mansoni-infected mice.
Authors: Chensue S W , Wellhausen S R , Boros D L
Published: December 2022

The regulatory mechanism(s) active in the spontaneous suppression of parasite egg-induced granulomatous response was analyzed in Schistosoma mansoni-infected mice. A single injection of cyclophosphamide (CY) given to chronically infected mice with diminished granulomas and impaired ability to produce MIF restored the enhanced granulomatous response and lymphokine production by spleen cells. Drug-treated animals also showed a rise in the level of circulating anti-egg antigen antibody. After CY treatment, spleen cells were still capable of adoptively suppressing the vigorous granulomatous response and MIF production in acutely infected recipients. Adoptive suppression, however, was abrogated when spleen cells of chronically infected mice were pretreated with anti-Lyt 1.1 alloantiserum and C before transfer. Based on the present and previous observations, it is proposed that the modulated granulomatous inflammatory response in mice with chronic infection is maintained by a dynamic equilibrium between effector and regulator lymphocytes. Although the inflammation is maintained by Ly 1+ Ia- TDH cells, the intensity of the response is regulated by Ly 1+ Ia+ TH, Ly 2+ Ia+ Ts and possibly a putative precursor population of lymphocytes.
Studies on Paul-Bunnell (P-B) antigen-antibody system. II. P-B antigens in extracts of lymphoma-leukemia spleens and pathologic sera.
Authors: Masaki M , Kano K , Merrick J M , Milgrom F
Published: December 2022

Preparations obtained by the chloroform-methanol extraction procedure from spleen tissues of patients with Hodgkin's disease, lymphomas, and leukemias, as well as from peripheral blood buffy coat of infectious mononucleosis (IM) patients were studied for the presence of 2 Paul-Bunnell (P-B) antigens; BS antigen shared by bovine red blood cells (BRBC) and sheep red blood cells (SRBC) and another, B antigen characteristic for BRBC. Both BS and B antigens were demonstrated by means of agglutination inhibition tests in over 40% of these extracts. None of the extracts from spleens, tonsils, and buffy coat of apparently normal human beings contained these antigens. P-B antigens of lymphoma-leukemia extracts were further purified by DEAE-Sephadex column chromatography. The purified fractions of some of these spleen extracts formed a precipitation line with IM sera, which merged into a reaction of identity with the lines formed by P-B antigens of BRBC. In studying various pathologic sera, B antigen was detected in sera of 28% of lymphoma-leukemia patients, 15% of patients with carcinomas of internal organs, and 3% of patients with systemic lupus erythematosus. On the other hand, BS antigen was found in only 3% of lymphoma-leukemia sera. These results confirmed our previous observations and indicated that both BS and B antigens are expressed as neoantigens on the patient's spleen cells as a result of pathologic processes in lymphoreticular malignancies.
Lectin(s) extracted from seeds of Artocarpus integrifolia (jackfruit): potent and selective stimulator(s) of distinct human T and B cell functions.
Authors: Bunn-Moreno M M , Campos-Neto A
Published: December 2022

A lectin activity that selectively induces different functions of human lymphocytes has been described in a PBS crude extract obtained from the seeds of Artocarpus integrifolia (jackfruit). Both unfractionated peripheral blood mononuclear cells and purified T cells are strongly stimulated to proliferate by this extract, whereas purified B cells are not. However, the lectin induced a potent polyclonal activation of B cells measured by a reverse hemolytic plaque assay using a multivalent anti-human Fab antibody.
HLA-A,B antigens Ia-like antigens, and tumor-associated antigens on prostate carcinoma cell lines: serologic and immunochemical analysis with monoclonal antibodies.
Authors: Ng A K , Pellegrino M A , Imai K , Ferrone S
Published: December 2022

Serologic and immunochemical analysis of the antigenic profile of the 2 human prostate carcinoma cell lines DU-145 and H494 with a battery of monoclonal antibodies has shown that both cell lines express HLA-A,B alloantigens and the 94,000 m.w. tumor-associated glycoprotein recognized by the monoclonal antibody 376.96S. In addition, the cell line H494 unexpectedly expresses Ia-like antigens, which are similar in their antigenic profile and structure to B lymphoid cell derived Ia-like antigens. Both Ia-like antigens and tumor-associated antigens can function as targets of cell-dependent lysis mediated by the corresponding monoclonal antibodies.
Lymphocytes bearing Fc receptors for IgE. VI. Suppressive effect of glucocorticoids on the expression of Fc epsilon receptors and glycosylation of IgE-binding factors.
Authors: Yodoi J , Hirashima M , Ishizaka K
Published: December 2022

Incubation of rat lymphocytes with homologous IgE induced an increase in Fc epsilon R(+) lymphocytes and the formation of IgE-binding factors. Pretreatment of rat lymphocytes with 1 to 5 microM dexamethasone, however, prevented the IgE-induced expression of Fc epsilon R on both B and T lymphocytes. Upon incubation with IgE, T cells activated with 10 micrograms/ml Con A produced IgE-potentiating factors that had affinity for lentil lectin and selectively enhanced the IgE response. Pretreatment of the Con A-activated T cells with dexamethasone, before incubation with IgE, changed the nature of IgE-binding factors formed by the cells. The majority of IgE-binding factors formed by the dexamethasone-treated, Con A-activated cells failed to bind lentil lectin Sepharose and selectively suppressed the IgE response. An injection of 0.2 mg dexamethasone into rats infected with Nippostrongylus brasiliensis markedly diminished the proportion of Fc epsilon R(+) lymphocytes in their mesenteric lymph nodes. The lymph node cells from the infected animals spontaneously released IgE-potentiating factors in vitro. However, the majority of IgE-binding factors formed by the mesenteric lymph node cells from the dexamethasone-treated, Nb-infected animals lacked affinity for lentil lectin and selectively suppressed the IgE response. The results indicate that glucocorticoid treatment prevented the glycosylation of IgE-binding factors and thereby changed the biologic activities of the factors.
Membrane sialoglycolipids emerging as possible signal transducers for lymphocyte stimulation.
Authors: Spiegel S , Wilchek M
Published: December 2022

Biotin hydrazide was attached covalently to the aldehyde groups produced by periodate oxidation of bovine brain gangliosides. These modified gangliosides were incorporated into mature rat thymocytes by incubation of the biotinyl gangliosides in the culture medium containing these cells. Avidin, which binds strongly to biotin, agglutinated and stimulated DNA synthesis in thymocytes containing the incorporated biotin-tagged gangliosides. Avidin has no mitogenic effect on normal thymocytes or on cells that incorporate unmodified gangliosides. With fluoresceinated avidin, the incorporated biotinyl gangliosides are shown to be laterally redistributed into patches and caps. These results imply that gangliosides may be involved in transmembrane communication during lymphocyte stimulation.
Antigen-specific helper factors present in the supernatant of concanavalin A-induced spleen cell cultures.
Authors: Kilburn D G , Anaka R
Published: December 2022

The supernatants from cultures of concanavalin A-induced spleen cells contained both antigen-specific and nonspecific (Interleukin 2) helper factors (Hf). The antigen-specific factor could be isolated from the supernatant by adsorption onto and elution from antigen-Sepharose immunoadsorbents. Specific Hf was produced in cultures of either immune or nonimmune spleen cells although in the latter case the quantity of Hf was significantly less. The specific Hf did not manifest the thymocyte stimulatory property of 112.
The parallel, time-dependent, bimodal change in lymphocyte cholinergic binding activity and cholinergic influence upon lymphocyte-mediated cytotoxicity after lymphocyte activation.
Authors: Strom T B , Lane M A , George K
Published: December 2022

The potent muscarinic cholinergic antagonist 3-quinuclidinyl benzilate has been used to detect muscarinic acetylcholine receptors on rodent and human lymphocytes. Binding to B lymphocytes was minimal and was not saturable or ligand specific. Half-maximal binding of (3H)-quinuclidinyl benzilate to T lymphocytes occurred at concentrations comparable to those described in other systems, and was displaceable at physiologic concentrations by muscarinic but not nicotinic agonists and antagonists. Binding to T lymphocytes was both saturable and specific, and was found to rapidly increase significantly after mitogen activation. Activation of T lymphocytes or the Lyt 1+ subset produces an early increase and later fall in muscarinic binding, and the ability od cholinergic agonists to augment the effector function of cytotoxic T lymphocytes harvested from alloimmune rat spleen is directly related to the magnitude of muscarinic binding.
Activation of macrophages for tumor cell cytotoxicity: identification of indomethacin sensitive and insensitive pathways.
Authors: Drysdale B E , Shin H S
Published: December 2022

Macrophages can be activated for tumor cell cytotoxicity by endotoxin- or lymphokine-containing solutions, and in both cases activation can be blocked by the addition of indomethacin. In contrast, the activation of macrophages by nonadherent or inflammatory peritoneal cells or antibody-coated tumor cells is not affected by indomethacin. These results demonstrate that there are at least 2 distinct pathways of macrophage activation, only 1 of which is affected by the addition of the drug. Activation by endotoxin that is indomethacin sensitive requires 2 steps. At present, the first is undefined and not blocked by indomethacin, but the second is inhibited by the drug and appears to require the production of E series prostaglandins. Our results also suggest that neither step alone is sufficient to activate macrophages for cytotoxicity.
Amyloidosis in (CBA/J X A/J)F2 mice: correlation of amyloid resistance and low mitogenic response to concanavalin A.
Authors: Wohlgethan J R , Cathcart E S
Published: December 2022

A/J mice are highly resistant to induction of amyloidosis with serial injections of azocasein, compared to the CBA/J strain. (CBA X A) F2 hybrid mice were treated with azocasein, after spleen biopsy for assay of 3 immunologic functions differing in the parent strains. The proportions of amyloid susceptibility and resistance in the hybrid population conformed to the expectations for a single gene. Low responses to the T cell mitogen concanavalin A correlated with resistance to amyloid induction, whereas the response to lipopolysaccharide and the level of natural killer activity were independent of susceptibility to amyloidosis.
A differentiation antigen of human NK and K cells identified by a monoclonal antibody (HNK-1).
Authors: Abo T , Balch C M
Published: December 2022

A monoclonal IgM antibody (HNK-1) was produced against a membrane antigen from the cultured T cell line, HSB-2. By indirect immunofluorescence, this antibody reacted with certain human cultured T cell lines (HSB-2 and MOLT-4 but not MOLT-3) but not with other lines of B cell or phagocytic cell origin. HNK-1 reacted with 15.1 +/- 7.1% of normal blood lymphocytes but was unreactive with monocytes, granulocytes, erythrocytes, and platelets. HNK-1+ cells separated by a fluorescence-activated cell sorter (FACS) were a homogeneous population of medium sized lymphocytes with abundant neutrophilic cytoplasm containing azurophilic granules. HNK-1+ cells were nonadherent, surface Ig-, mostly FcIgG receptor+ and both positive and negative for demonstrable sheep erythrocyte (E) rosetting capability. Cell suspensions enriched for E-rosetting T cells and depleted of FcIgG receptor+ cells contained few (6%) HNK-1+ cells. Depletion of HNK-1+ cells from blood mononuclear cell populations by complement (C) mediated lysis greatly reduced NK activity against K-562 target cells and K cell lytic activity against antibody-coated chicken red blood cells. Treatment with HNK-1 alone or C alone did not affect these activities. When the FACS was utilized to separate HNK-1+ and HNK-1- cells from 6 individuals, the HNK-1+ cell population contained almost all of the NK and K cell function. The monoclonal antibody HNK-1 thus defines the first differentiation antigen shown to be selectively expressed on human NK and K cells.
Induction of complement receptor expression in cell lines derived from human undifferentiated lymphomas. II. Characterization of the induced complement receptors and demonstration of the simultaneous induction of EBV receptor.
Authors: Magrath I , Freeman C , Santaella M , Gadek J , Frank M , Spiegel R , Novikovs L
Published: December 2022

We have studied the specificity of complement receptors induced by theophylline in 2 cell lines derived from undifferentiated lymphomas, one of Burkitt's type, and compared it to that of complement receptors in other cell types. Both C3b and C3d receptors were induced. The induced C3b receptor differed from the C3b receptor of mature normal lymphocytes, polymorphonuclear leukocytes and the cells of a nodular lymphoma in 2 respects. Firstly, it bound C3b much less avidly (by a factor of several hundred-fold) and secondly, we were unable to demonstrate C4b binding. EBV receptors were induced at the same time as complement receptors, and permitted the conversion of a greater fraction of cells to EBNA positivity after experimental infection with EBV. The induction of receptors was not associated with a change in the fluidity of the plasma membranes and our data do not favor a different orientation of induced receptors within the membrane as compared to receptors of other cell types--a potential explanation for the different specificities. Our findings are consistent with the possibility that the complement receptors of lymphocyte precursors differ from these of mature lymphocytes.
Order of events leading to surface immunoglobulin capping: analysis of a transmembrane signal.
Authors: Young-Karlan B R , Ashman R F
Published: December 2022

Capping provides a rapid assay for the transduction of 1 type of membrane signal generated by the cross-linking of cell surface receptors. The order of the steps comprising this signal was determined by employing reversible inhibitors of lymphocyte surface Ig capping in a sequential incubation protocol. The results demonstrated that surface Ig cross-linking leads to capping by a linear series of discrete events. Although the steps inhibited by the calcium ionophore A23187 and the tranquilizer chlorpromazine could not be distinguished, other agents also thought to influence calcium distribution in the cell acted at different steps. The order of inhibited steps was shown to be: 1) hydrocortisone; 2) calcium ionophore or chlorpromazine; 3) cytochalasins; 4) dibucaine; 5) propranolol; 6) fluoride; 7) azide. These results suggest a model wherein cross-linked membrane Ig aggregates engage the preassembled microfilament system by means of a calcium-dependent linkage. Further calcium redistribution within the cell then leads to an energy-consuming contractile event.
Ontogeny and postnatal persistence of a strong suppressor activity in man.
Authors: Unander A M , Olding L B
Published: December 2022

We report here on the ontogeny and postnatal persistence of an inhibited human immune response in which lymphocytes from human newborns strongly suppress the proliferation of adults' lymphocytes stimulated by phytohemagglutinin (PHA) or alloantigens in vitro. For this research we used a 2-way mixed lymphocyte culture (MLC) supplemented with PHA, with sex chromosomes acting as markers for dividing male and female cells, or alternatively a double chamber system. The proliferation of maternal lymphocytes was significantly suppressed by fetal lymphoid cells from the liver as early as the 8th week of gestation and by those from fetal blood at the 14th week or later during gestation. This strong suppressor activity persisted in 11-mo-old infants but usually disappeared after that time.
Sequential production of IFN-alpha and immune-specific IFN-gamma by human mononuclear leukocytes exposed to herpes simplex virus.
Authors: Green J A , Yeh T J , Overall J C
Published: December 2022

Both viral-induced interferon (IFN-alpha) and immune interferon (IFN-gamma) were produced in cultures of human peripheral mononuclear leukocytes (PML) exposed to ultraviolet (UV) light-inactivated herpes simplex virus (HSV). Comparable IFN titers (100 to 300 U) were present in supernatant media from PML of HSV-seropositive and -seronegative donors maintained for 2 days in culture. This "early" IFN was not immunologically specific and had the characteristics of IFN-alpha. In contrast, PML from 20 of 21 (95%) seropositive donors, but only 2 of 22 (9%) seronegative subjects, produced IFN in day-5 cultures that had been washed on day 2 followed by the addition of fresh media without UV-HSV (p < 0.0001). In addition,the geometric mean IFN titer of 35 U present in media from day 5 cultures of seropositive donors was significantly higher (p < 0.0001) than those from seronegative subjects (<10 IFN U). This "late" IFN was defined as immunologically specific by virtue of its being present almost exclusively in PML cultures from seropositive donors. Late-appearing, HSV-induced, immunospecific IFN-gamma had antigenic characteristics of both viral-induced IFN-aand mitogen-induced IFN-gamma. A considerable degree of overlapping sensitivity to anti-IFN-alpha and anti-IFN-gamma antibody existed in late IFN, but was not seen in standard, control specimens. The latter finding suggests that the possible existence of an IFN of antigenic character intermediate to IFN-alpha and IFN-gamma. Late IFN was not induced in PML cultures by UV-inactivated Newcastle disease virus (NDV). NDV did, however, induce high titers of early IFN-alpha in PML cultures of both HSV-seropositive and HSV-seronegative donors. HSV-stimulated lymphocyte blastic transformation occurred only in cultures of seropositive individuals, whereas none occurred in response to NDV. Delayed appearance of antiviral activity with the characteristics of IFN-gamma in HSV-stimulated PML cultures is a sensitive and specific measure of the human immune IFN response. Sequential synthesis of IFN-alpha and IFN-gamma in individual PML cultures provides an opportunity to examine regulation of IFN production in response to virus with intrinsic IFN-inducing properties.
A mouse IgM allotypic determinant (Igh-6.5) recognized by a monoclonal rat antibody.
Authors: Kung J T , Sharrow S O , Sieckmann D G , Lieberman R , Paul W E
Published: December 2022

Two monoclonal rat anti-mouse IgM antibodies, Bet 1 and Bet 2, are described in this paper. Bet 1 defines a new allotypic determinant, Igh-6.5, expressed on IgM molecules in serum and on B lymphocytes, whereas Bet 2 recognizes a determinant on IgM molecules of all mouse strains tested. Both reagents bind to the IgM myeloma protein MOPC104E, but not to IgG myeloma proteins, including FLOPC21, MOPC21, and UPC10. Using serum from various mouse strains to inhibit the binding of Bet 1 to MOPC104E, 3 distinct inhibition patterns were found. BALB/c, DBA/2, and CBA sera inhibited strongly, C56BL/6 (B6), SJL, AKR, and NZB sera inhibited weakly, and A and AL sera showed no inhibition of binding of BET 1 to MOPC104E. All sera tested were equivalent in their inhibition of the binding of Bet 2 to MOPC104E. When spleen cells from different mouse strains were reacted with fluorescein-conjugated Bet 1 (F-Bet 1) and subjected to flow microfluorometry analysis, 3 types of staining patterns, corresponding to those obtained with the serum inhibition assay, were also found. The determinant recognized by Bet 1 is controlled by a gene linked to the Igh-C gene complex. C.AL20 behaved like AL, and C.B20 and BAB/14 behaved like B6, both in the serum inhibition assay and in flow microfluorometry analysis of spleen cells stained with F-Bet 1. In addition, the capacity of serum from individual (BALB/c X B6) X B6 backcross progeny to inhibit the binding of Bet 1 to MOPC104E was linked to the expression of the Ig-1a marker.
Changes in lectin receptor lateral mobilities accompany lymphocyte stimulation.
Authors: Leuther M D , Peacock J S , Krakauer H , Barisas B G
Published: December 2022

Changes in lateral mobilities of rabbit lymphocyte membrane components in response to succinyl concanavalin A (S Con A) have been studied by fluorescence photobleaching recovery (FPR). During hrs 0 to 3 after exposure to S Con A, lectin receptor mobilities on both T and B cells fall about 2-fold. Reduced mobility of T cell lectin receptors persists until hr 18. From hr 18 to 24 rapid recovery of original mobility occurs if and only if lectin is present. In contrast, nonresponding B cells recover original receptor mobility gradually over hr 4 to 48. Metabolic inhibitors added at hr 3 restore original receptor mobilities, but cytoskeletal disruptors have this effect on T cells only. From hr 0 to 15, washing lectin from the cell surface is decreasingly effective in restoring T cell receptor mobility. After hr 15, mobility cannot be enhance by lectin removal. Parallel DNA synthesis studies show that, for T cell stimulation, lectin must be present on the cell surface during hr 0 to 3 and 18 to 24. These are the periods when FPR measurements show lectin receptor mobilities being restricted and released, respectively. Stimulation of B cells by anti-Ig shows several interesting features. First, stimulation by intact anti-Ig fails to reduce the mobilities of Con A receptors in a manner similar to that produced by S Con A. Second, S Con A does reduce mobility of surface Ig. Thus, Con A receptors would appear to exert a unique anchorage modulation of mobilities of other membrane molecules.
Induction of immunoglobulin-secreting cells in the allogeneic mixed leukocyte reaction: regulation by helper and suppressor lymphocyte subsets in man.
Authors: Kotzin B L , Benike C J , Engleman E G
Published: December 2022

The existence of functionally distinct T lymphocyte subsets in man, initially demonstrated with heteroantisera, has been confirmed with monoclonal reagents. Two major subsets have been defined: Leu-2 cells, which effect cell mediated lymphocytotoxicity (CML), and Leu-3 cells, which amplify CML and other T cell functions. This study is an effort to determine the effects of these subsets on the immunoglobulin secretion induced in the human mixed leukocyte reaction (MLR). T cells were separated from non-T cells by rosetting with sheep erythrocytes and were fractionated into Leu-2 and Leu-3 subsets by solid phase immunoabsorption with monoclonal antibodies. T cell subsets were cultured with autologous non-T cells and irradiated allogeneic stimulator non-T cells. Secretion of IgM and IgG was measured by a reverse hemolytic plaque assay. MLR-induced antibody secretion was specifically dependent on the Leu-3 T lymphocyte subset. The Leu-2 subset was incapable of generating large numbers of IgM- or IgG-secreting cells, and, in fact, suppressed the Leu-3-induced response. Exposure of Leu-3 cells to a dose of irradiation sufficient to prevent their proliferation in MLR did not reduce induced immunoglobulin secretion. Leu-2-mediated suppression, however, was sensitive to low dose irradiation. Thus Ig secretion in human MLR is regulated by a balance of helper activity from the Leu-3 subset and suppressor activity from the Leu-2 subset.
Inhibition of IgE-mediated histamine release from rat basophilic leukemia cells and rat mast cells by inhibitors of transmethylation.
Authors: Morita Y , Siraganian R P
Published: December 2022

This study evaluated the effect of inhibitors of transmethylation on histamine release from rat mast cells and rat basophilic leukemia cells. IgE-mediated histamine release from rat basophilic leukemia cells (RBL-2H3 cells) was inhibited by 3-deazaadenosine (DZA) in the presence of L-homocysteine thiolactone (Hcy) or the combination of adenosine, erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA), and Hcy in a dose-dependent fashion. There were no significant changes in the cellular cAMP levels by these inhibitors. Histamine release induced by anti-IgE or dextran from normal rat mast cells was also blocked by DZA plus Hcy in a dose-dependent manner. DZA at 10(-3) M in the presence of 10(-4) M Hcy or the combination of 10(-3) M adenosine, 10(-4) M EHNA, and 10(-3) M Hcy inhibited lipid (perhaps phospholipid) methylation into RBL-2H3 cells without affecting choline incorporation. In the presence of 10(-3) M DZA plus 10(-4) M Hcy there was a 170-fold increase in [35S]AdoHcy with the concomitant appearance of 3-deaza-AdoHcy when the cells were incubated with [35S]methionine, thus indicating that these drugs inhibited methylation reaction(s) through the intracellular accumulation of AdoHcy and 3-deaza-AdoHcy. In contrast, histamine release from rat mast cells induced by the calcium ionophore A23187, compound 48/80, polymyxin B, or ATP was not inhibited by these compounds. These results suggest that IgE- or dextran-mediated histamine release involves methylation reactions(s), whereas the other secretagogues bypass this early step.
Characterization of the human complement (c3b) receptor with a fluid phase C3b dimer.
Authors: Arnaout M A , Melamed J , Tack B F , Colten H R
Published: December 2022

The interaction of C3b receptor with C3b, the major cleavage product of C3, elicits important biologic functions, such as enhanced phagocytosis and release of cellular enzymes. We determined the binding kinetics and binding isotherm of C3b-receptor interaction by using human cells and fluid phase C3b generated by trypsin cleavage of purified native C3. 125I labeled C3b was separated into 2 molecular species, a dimer and a monomer by column chromatography. We found that dimeric C3b bound to human erythrocyte C3b receptors with an affinity that was more than 25 times that of the monomer. 125I dimeric C3b did not bind to sheep red blood cells, which lack the C3b receptor, nor to trypsinized or 2-mercaptoethanol treated normal human red blood cells, 2 methods for abrogating the immune adherence activity. Binding of 125I fluid phase C3b dimer to the C3b receptor was specific, saturable (about 90 ng of C3b dimer bound maximally per 1 X 10(9) red blood cells), reversible (in the presence of a 100-fold molar excess of unlabeled ligand), and of moderate affinity (Kd about 9.53 nM). The equilibrium binding constants were similar with the various cells tested. Binding was characterized by rapid on and off rates and did not exhibit ligand cooperativity. This specific interaction reached a steady state within 10 to 15 min at 0 degrees C; 50% of specifically bound ligand dissociated from its binding site on human red blood cells in approximately 1 min at 0 degrees C. The density of C3b receptors on human red blood cells, polymorphonuclear leukocytes, monocytes, and B lymphocyte-enriched preparations was 360, 20,000, 30,000, and 21,000 receptors/cell, respectively.
A new possible regulatory system for protein phosphorylation in human peripheral lymphocytes. II. Possible relation to phosphatidylinositol turnover induced by mitogens.
Authors: Ku Y , Kishimoto A , Takai Y , Ogawa Y , Kimura S , Nishizuka Y
Published: December 2022

Ca++-activated, phospholipid-dependent protein kinase present in human peripheral lymphocytes requires a small amount of diacylglycerol in addition to phospholipid, particularly at lower concentrations of Ca++. It is necessary that such diacylglycerol contain unsaturated fatty acid at least at position 2. Saturated diacylglycerols such as dipalmitin and distearin are far less effective. Kinetic analysis indicates that unsaturated diacylglycerol greatly increases the apparent affinity of the enzyme for phospholipid, and sharply decreases the Ca++ concentration to the micromolar range that gives rise to the maximum enzyme activation. Among various phospholipids tested, phosphatidylserine is most active in supporting enzymatic activity. Phosphatidylinositol and phosphatidylethanolamine are less effective. Phosphatidylcholine, phosphatidic acid, sphingomyelin, and lysophosphatidylcholine are inert. It is most likely, therefore, that various lymphocyte mitogens induce specific hydrolysis of phosphatidylinositol to produce such an active unsaturated diacylglycerol, which in turn serves as a second messenger for the selective activation of this unique protein kinase. Dibucaine and chlorpromazine appear to interact with phospholipid and thereby inhibit the activation process of this enzyme. Cyclic nucleotide-dependent protein kinases are not susceptible to these phospholipid-interacting drugs.
Helper cells active in the generation of cytotoxicity to a syngeneic tumor.
Authors: Hancock E J , Kilburn D G , Levy J G
Published: December 2022

We have examined a population of thymic lymphocytes that augment the generation in vitro of specific cytotoxic T cells against P815.X2, a syngeneic tumor in DBA/2 mice. These helper cells are evident in the thymus of DBA/2 mice 5 to 7 days after subcutaneous injection of live P815 cells. They are Thy-1 positive, resistant to gamma radiation, antigen specific, and appear to exert their influence most effectively when added at the beginning of the culture.
Virus-immune and alloreactive response characteristics of thymocytes and spleen cells from young mice.
Authors: Schwartz D H , Doherty P C
Published: December 2022

Thymocytes from 3-day-old mice generate a strong, virus-immune cytotoxic T lymphocyte (CTL) response when primed in irradiated, vaccinia-infected recipients. Effector potential is not detected in spleen until day 4 or 5, when alloreactive precursors may also be present. Mice of 8 days or older survive inoculation with large doses of vaccinia virus, and generate significant CTL activity in spleen. There is, unlike the situation described by others for Listeria antigens, no major defect in stimulator function for spleens of 1-wk-old mice. This is true for both the virus-specific and alloreactive CTL responses.
Establishment of unresponsiveness in primed B lymphocytes in vivo.
Authors: Nelson-Rampy P A , Parks D E , Weigle W O
Published: December 2022

As an approach to examine the influence of the state of cellular activation on the ability to tolerize B cells, the induction of unresponsiveness in human gamma-globulin-(HGG) primed B lymphocytes was studied in an adoptive transfer system. In contrast to transferred normal spleen cells, spleen cells from HGG-primed mice are not readily rendered unresponsive when exposed to the tolerogen, deaggregated HGG (DHGG), in irradiated recipients. A kinetic study showed that unfractionated primed spleen cells do not respond to an antigenic challenge given between 6 and 10 days after cell transfer and injection of DHGG, indicating that they are transiently depressed. In contrast, isolated primed B cells are tolerized when transferred to recipients and treated with DHGG in the absence of T cells. Furthermore, primed B cells exposed to tolerogen in the recipients do not recover the ability to respond to HGG either after a secondary challenge with AHGG given up to 14 days after transfer, or after 2 consecutive challenges given on days 14 and 24 after transfer. The presence of primed T cells at the time of tolerization interferes with the induction of unresponsiveness in these primed B cells. These studies suggest that the presence of primed T cells is responsible for the inability to tolerize unfractionated primed spleen cells populations and that primed B cells themselves are not intrinsically resistant to the induction of unresponsiveness.
A C1-inhibitor-complex assay (INCA): a method to detect C1 activation in vitro and in vivo.
Authors: Hack C E , Hannema A J , Eerenberg-Belmer A J , Out T A , Aalberse R C
Published: December 2022

A radioimmunoassay (the C1-inhibitor-complex assay, INCA) is described for the detection of complexes that are composed of at least C1s and C1-inhibitor. This INCA is based on demonstrating that C1s and C1-inhibitor (C1-In) are linked: after an incubation with anti-C1s-Sepharose, bound C1sC1-In complexes are detected by 125I-anti-C1-In. C1sC1-In complexes were prepared by the addition of a slight excess of C1s to normal human serum (NHS). As little as 2 ng C1-In bound to C1s was detected. Additional free C1s in serum hardly influenced the detection of C1sC1-In complexes. Complexes presumably composed of C1rC1s(C1-In)2 were generated by the addition of aggregated IgG to NHS. This generation was inhibited by lowering the temperature to 0 degrees C, and by EDTA, and depended on the concentration of aggregated IgG. These complexes had a sedimentation value of approximately 9S. Complexes of C1s and C1-In were also generated in NHS by the addition of DNP-albumin and protein A, but not by zymosan. The INCA was applied to blood samples from normal donors and patients. Sixteen out of 19 samples from patients with acute glomerulonephritis contained increased amounts of C1rC1s(C1-In)2 complexes as compared with the amounts in blood samples from normal donors. The INCA provides a useful tool to assess the activation of C1 in the presence of C1-In, both in vitro and in vivo.
Differential glycosylation of murine B cell and spleen adherent cell Ia antigens.
Authors: Cullen S E , Kindle C S , Shreffler D C , Cowing C
Published: December 2022

Examination of I-A and I-E molecules from purified B cells and splenic adherent cells from various haplotypes revealed a consistent difference in isoelectric focusing (IEF) that has been localized to the alpha-chain. alpa-Chains from B cells appeared heterogeneous and contained acidic IEF bands absent from the adherent cell I-A molecules. No difference in Ia beta-chain or H-2 IEF patterns was observed when B cell and adherent cell preparatios were compared when B cell and adherent cell preparations were compared. I-A alpha-chain preparations from the 2 cell sources showed no differences in 3H-leucine-labeled tryptic peptides separated by reverse-phase high-pressure liquid chromatography. Digestion with neuraminidase, kinetics of labeling, and subcellular distribution indicated that the extra acidic IEF bands in B cell Ia represent a mature, more heavily sialated form of alpha-chain that is not present in adherent cells. The selective differential glycosylation of B cell and adherent cell Ia could have some relation to the function of those cell types or to cell type-specific recognition of those cells.
The metabolic relationships of the immunoglobulins in fish serum, cutaneous mucus, and bile.
Authors: Lobb C J , Clem L W
Published: December 2022

Passive administration of homologous, radioiodinated, purified high m.w. (HMW) or low m.w. (LMW) serum immunoglobulin (Ig) into the sheepshead, a marine teleost, showed that the LMW Ig was neither an extracellular precursor nor a degradation product of the HMW Ig. These studies also showed that the half-lives of the HMW and LMW Ig were statistically identical, i.e., approximately 16 days. Furthermore, this work demonstrated that the Ig found in cutaneous mucus and bile were not due to transudation or active transport of either the serum HMW or LMW Ig. The implications of these later findings, coupled with previous physicochemical characterizations of the Ig in fish secretions, support the hypothesis that fish, similar to higher vertebrates, have a secretory immune system.