bluuebunny/crossref_metadata_2025 huggingface dataset:
Relative resistance of the F-42-stabilized classical pathway C3 convertase to inactivation by C4-binding protein.
Authors: Daha M R , van Es L A
Published: December 2022

The sera of some patients with SLE contain an IgG antibody (F-42) directed against the classical pathway C3 convertase (C-42), which is capable of stabilizing C42 in a dose-dependent manner. The half-life (T 1/2) of C42 is prolonged by F-42. In order to determine whether C4-binding protein was capable of reversing stabilization of C42, stabilized and unstabilized cell-bound C42 were exposed to purified C4-bp and the convertase activity was assessed. C4-bp was capable of accelerating the decay of C42 in a dose-dependent manner; 2 microgram/ml C4-bp reduced the T 1/2 of C42 from 5 to 2.5 min at 30 degrees C. On the other hand, 16 microgram C4-bp could reverse stabilization of C42 by F-42 from T 1/2 = 78 min to a T 1/2 - 40 min; 128 microgram C4-bp reduced the T 1/2 of stabilized C42 to 4 min. Functional inactivation of C42 occurs via enhanced decay-dissociation of C2 from the convertase by C4-bp, as shown by the release of 125I-C2i from the cell-bound convertase. Stabilization of C42 by F-42 is caused by prevention of decay-dissociation of 125I-C2. F-42 was also capable of stabilizing C4oxy2 even further, as shown by prolongation of the T 1/2 of cell-bound C4 oxy2 to a T 1/2 of at least 300 min at 30 degrees C.
Regulation of lymphocyte responses by human gangliosides. I. Characteristics of inhibitory effects and the induction of impaired activation.
Authors: Whisler R L , Yates A J
Published: December 2022

Gangliosides obtained from normal human brain were found to inhibit the in vitro activation of human lymphocytes by nonspecific mitogens and allogeneic cells at concentrations between 3 to 50 microgram/1.5 to 1.7 X 10(5) lymphocytes/0.2 ml culture. Ganglioside inhibition did not represent cytotoxic effects or altered lectin binding and was independent of the mitogen concentration. In addition to concentration, the degree of inhibition was dependent on the mode of presentation to lymphocytes, since gangliosides incorporated within liposomal membranes displayed a synergistic inhibitory effect greater than predicted from the cultures receiving either gangliosides or liposomes alone. In binding experiments, radiolabeled ganglioside GM1 became associated with human lymphocytes within 10 min. However, approximately 72 hr pre-exposure of human lymphocytes to gangliosides was required to induce impaired lymphocyte responses to mitogens and allogeneic cells. Thus, concentrations of human gangliosides equivalent to the levels occurring in the sera of patients with certain malignancies are capable of actively inhibiting lymphocyte stimulation in addition to inducing impaired lymphocyte responses.
Regulatory role of IgE-binding factors from rat T lymphocytes. IV. Formation of IgE-binding factors in rats treated with complete Freund's adjuvant.
Authors: Hirashima M , Yodoi J , Ishizaka K
Published: December 2022

Mesenteric lymph node (MLN) cells from rats treated with complete Freund's adjuvant (CFA) spontaneously released soluble factors, which have affinity for IgE (IgE-binding factors). The factors were purified by absorption of culture filtrates of the lymphocytes with IgE-Sepharose followed by elution at acid pH. Purified IgE-binding factors selectively suppressed IgE-forming cell response of DNP-ovalbumin-primed MLN cells to homologous antigen without affecting the IgG2 response. Fractionation of purified IgE-binding factors on lentil lectin-Sepharose showed that the majority of IgE-binding factors did not have affinity for lentil lectin, whereas a minor fraction was bound to the lectin-coupled beads. It was found that IgE-binding factors lacking affinity for lentil lectin suppressed IgE response, whereas those eluted from lentil lectin-Sepharose selectively enhanced IgE response. Serum of CFA-treated animals also contained both IgE-suppressive factor and IgE-potentiating factor, and the amount of the former factor was more than the latter. The source of IgE-suppressive factor is non-B lymphocytes. Depletion of Fc, receptor(+) lymphocytes in MLN cells from CFA-treated animals did not affect the formation of IgE-suppressive factor, indicating that Fc epsilon R-bearing cells are not the source of the factor.
Mechanism of lethal effect of human serum upon Leishmania donovani.
Authors: Pearson R D , Steigbigel R T
Published: December 2022

In order to gain greater understanding of potential host defense mechanisms against Leishmania donovani, we examined the effect of nonimmune, human serum upon promastigotes and amastigotes. Fresh sera were found to be lethal for promastigotes, but had no detectable effect on amastigotes. Serum exposed promastigotes became immotile, did not take up neutral red dye, appeared to be disrupted, and failed to recover after further incubation in fresh media. Heat labile components were required for promastigote killing since heat-inactivated serum (56 degrees C, 30 min) agglutinated but did not kill them. Sera that lacked either the 5th or 6th complement (C) component had no effect when used alone, but when used together, were lethal, indicating that activation of the membrane attack complex (C5b-C9) ws necessary for the lethal effect. The mode of C activation was determined by using serum with complete, selective, deficiency of C2, and normal serum chelated with Mg-EGTA. The C2-deficient serum killed promastigotes only after the addition of purified C2, and Mg-EGTA chelated serum had no detectable lethal effect. Thus, promastigotes appeared to activate C through the classical pathway. Human IgG and IgM, detected with 125I-anti-human antibody, bound to promastigotes. Removal of antibody from serum by absorption with promastigotes eliminated the lethal effect. The effect was restored by addition of heat-inactivated serum to absorbed serum. We conclude that promastigotes bind antibody and are killed by activation of the membrane attack complex of C through the classical pathway.
Lymphocyte activation by purified carrageenan.
Authors: Ishizaka S , Hasuma T , Otani S , Morisawa S
Published: December 2022

The induction of polyclonal antibody production and mitogenic response by purified carrageenan was studied in vitro by using spleen cells. It was shown that kappa-CGN, lambda-CGN, and iota-CGN induced significant responses in mice, hamsters, and rats. The less purified CGN also exhibited the polyclonal antibody production in mice. In addition, purified CGN exhibited mitogenic action on B cells and to a lesser extent, on thymocytes. Polyclonal antibody responses induced by purified CGN were not affected by removal of T cells, adherent cells, or iron powder phagocytic cells. These results suggest that CGN may activate B cells resulting in polyclonal antibody responses.
Inherited deficiency of the ninth component of complement in man.
Authors: Lint T F , Zeitz H J , Gewurz H
Published: December 2022

A 76-year-old man was found to have low (33% normal) serum complement (C) hemolytic activity, although C3 and C4 protein levels were normal. Further evaluation of his serum and plasma indicated that all C components were present in normal or elevated amounts except for C9, which was undetectable by both antigenic and functional assays. Addition of purified human C9 led to full restoration of the hemolytic activity. Family studies demonstrated that the deficiency was inherited as an autosomal codominant trait and was not linked with alleles at the HLA-A or HLA-B loci. The patient had no history of recurrent or unusual infections and no evidence of autoimmune disease. The availability of serum totally lacking in C9 permitted an investigation of the lytic capacity of the C5b-8 segment of the C attack mechanism, which was pursued in kinetic studies on the hemolysis of erythrocyte intermediates. These studies indicated that hemolysis occurred approximately 100 times slower in patient than in normal serum, using either EA or EAC1-7 intermediates as target cells. Serum bactericidal activity also was slower in patient serum, occurring at a rate about 1/35 that observed in normal serum. These studies provide direct independent evidence that cytolysis of erythrocytes and bacteria can be mediated by C5b-8, and allow a quantitative estimation of the increment in the rates of these reactions provided by normal serum levels of C9. The presence of readily detectable though slow hemolytic activity of C9-deficient serum may account for the difficulty in identifying individuals with this defect.
The role of agonists that activate adenylate cyclase in the control of cAMP metabolism and enzyme release by human polymorphonuclear leukocytes.
Authors: Marone G , Thomas L L , Lichtenstein L M
Published: December 2022

An inverse relationship between cAMP content and effector function is ascribed generally to immune and inflammatory cells. Previous reports imply, however, that human polymorphonuclear leukocytes (PMN) are less responsive than other inflammatory cells to adenylate cyclase (AC) agonists. We therefore examined the effects of isoproterenol, prostaglandin E1 (PGE1), adenosine, and histamine on the adenosine 3',5'-monophosphate (cAMP) content of PMN and on particle-stimulated lysosomal enzyme release. For comparison, the effect of AC agonists on the cAMP content of human peripheral lymphocytes was evaluated in parallel. Although potent stimuli for cAMP accumulation in lymphocytes, the AC agonists produced only marginal increases in the cAMP content of PMN; this difference in responsiveness was independent of agonist concentration or length of incubation. Inhibition of lysosomal enzyme release by the AC agonists was likewise marginal (< 20%). The addition of theophylline with isoproterenol produced additive inhibition without significant cAMP increases. Hydrocortisone, which caused a small increase in the cAMP content, markedly potentiated the effects of AC agonists on the cAMP level in PMN; the synergistic increases in cAMP were accompanied by additive effects on lysosomal enzyme release. It is concluded that human lymphocytes and PMN exhibit differential sensitivity to AC agonists and that this difference may provide a basis for the selective modulation of individual PMN- or lymphocyte-mediated events.
Differentiation of human B cells expressing the IgA subclasses as demonstrated by monoclonal hybridoma antibodies.
Authors: Conley M E , Kearney J F , Lawton A R , Cooper M D
Published: December 2022

Monoclonal hybridoma antibodies to the human IgA subclasses were produced by immunizing mice with purified myeloma proteins. These antibodies were shown to be specific for the appropriate IgA subclass by enzyme-linked immunoabsorbant assay (ELISA) and by immunofluorescent staining of myeloma plasma cells and B cells from normal individuals. These antibodies were used to demonstrate age-related shifts in the proportions of IgA1- and IgA2-bearing B cells that could be correlated with 3 distinct staining patterns. In the newborn equal numbers of IgA1 and IgA2, B cells were found. These cells had only small amounts of surface IgA in a patchy distribution. They also expressed surface IgM. In the infant, large lymphoblastoid cells were observed that bore more IgA in a homogeneous pattern but did not express IgM. Of these cells, 98% were positive for IgA1. In the adult, 80% of the IgA B cells were positive for surface IgA1, and 20% were positive for IgA2. These were small lymphocytes brightly stained for IgA and negative for IgM. In culture, the adult B cells responding to pokeweed gave rise to roughly equal numbers of IgA1 and IgA2 plasma cells. These results suggest that there are equal numbers of precursor cells for IgA1 and IgA2 whose expansion, further differentiation, and migration are selectively affected by immunoregulatory controls.
Differences in the regulation of humoral responses between mice infected with Trypanosoma cruzi and mice administered T. cruzi-induced suppressor substance.
Authors: Cunningham D S , Benavides G R , Kuhn R E
Published: December 2022

In the present study, the effects of Trypanosoma cruzi and the T. cruzi-induced serum suppressor substance (SSS) on antibody responses were compared. Although infection with T. cruzi led to an alteration in T cell helper activity and a reduced specific B cell precursor frequency, SSS did not have a similar effect on either of these cell populations. The characteristics of the altered T cell helper activity was further investigated, and it was found that helper activity appeared earlier in infected mice than in normal or SSS-suppressed mice, and less antigen was required for optimal elicitation of T helper cells in infected mice. The potency of T cell helper activity also was shown to differ, and in the order T. cruzi-infected 6E normal 6E SSS-suppressed mice. It was found that spleen cells from T. cruzi-infected mice elaborated more potent specific helper factors than spleen cells from normal or SSS-suppressed mice, but did not produce a detectable nonspecific helper factor in vitro. Finally, the addition of B cells from low-dose primed, T. cruzi-infected mice to cultures of normal spleen cells resulted in subnormal responses to the priming antigen (sheep erythrocytes) but not to another unrelated antigen (trinitrophenyl-haptenated Brucella abortus), whereas similarly sensitized B cells from normal or SSS-suppressed mice caused no such effect.
Clonal nature of the immune response to phosphocholine. IX. Heterogeneity among antibodies bearing M511 idiotypic determinants.
Authors: Wolfe J , Claflin J L
Published: December 2022

A fraction of serum anti-PC antibodies from mice immunized with PC-KLH expresses idiotypic determinants of the PC-binding myeloma MOPC-511. We examined the molecular heterogeneity of IgG antibodies bearing the M511 idiotype (Id) by analytical isoelectric focusing followed by in situ labeling with 125I-anti-M511. Antibodies with the M511 Id were shown to be conserved in many mouse strains and were distinct from T15 Id+ anti-PC antibodies. Two complex sets of M511 Id+ bands were observed and these were identified as M511 Id+ antibodies belonging to the IgG1 and IgG3 subclasses. Examination of responses in different individuals of a strain permitted identification of at least 4 to 6 IEF band groups. Thus, the M511 Id is a marker for a family of closely related molecules. Quantitative variations were observed in the expression of the M511 family within a strain and between strains of mice that occurred independently of expression of the dominant T15 Id+ fraction. These results support the concept of an M511 Id family and further suggest that there can be differential control mechanisms governing expression of individual clones or a family of closely related clones.
Synergistic activation by lymphokines and muramyl dipeptide of tumoricidal properties in rat alveolar macrophages.
Authors: Sone S , Fidler I J
Published: December 2022

Alveolar macrophages (AM) from lungs of normal F344 rats can be rendered tumoricidal by incubation in vitro with either muramyl dipeptide (MDP) at a minimum dose of 10 micrograms/ml or undiluted cell-free culture supernatants from mitogen-stimulated F344 rat lymphocytes rich in macrophage-activating factor (MAF) activity. Neither MAF at dilutions exceeding 1:6 nor MDP at doses lower than 10 micrograms/ml activated AM to become tumor cytotoxic. The combination of agents at subthreshold amounts (MAF 1:18; MDP 0.001 to 1 microgram/ml) activated AM to significant levels of cytotoxicity. AM activated by these agents were rendered tumoricidal and destroyed syngeneic, allogeneic, and xenogeneic tumor targets in vitro. The synergism for AM activation between preparations of MAF and MDP required that AM be incubated first with MAF and then with MDP. Even a 15-min treatment of AM with MAF conditioned the cells to respond to subthreshold amounts of MDP and to be rendered tumoricidal. Since treatment of MAF and MDP with polymyxin B did not interfere with macrophage activation, we were able to rule out the possibility that our preparations were contaminated with lipopolysaccharide. Synergism for AM activation was demonstrated also when AM were treated with MAF and MDP encapsulated within liposomes. This finding suggests that the binding of agents to the macrophage surface is not a prerequisite for the synergistic activation of AM by MAP and MDP.
Polyclonal activation of murine B lymphocytes by Fc fragments. III. Characterization of the defect in the ability of the C3H/HeJ mouse to respond to Fc fragments.
Authors: Morgan E L , Weigle W O
Published: December 2022

Spleen cell preparations from the C3H/HeJ mouse were found to be refractory to Fc fragment-induced polyclonal activation. This defect was not a characteristic of all C3H strains, because spleen cells from the histocompatible C3Heb/FeJ strain produced polyclonal antibody in the presence of Fc fragments. The cellular defect was found to be associated with the macrophage population. Macrophages from C3H/HeJ mice were incapable of generating mitogenic Fc subfragments from intact Fc, whereas C3Heb/FeJ performed this function normally. That T cells and B cells from the C3H/HeJ mouse were normal was shown by the fact that C3H/HeJ spleen cells responded to Fc subfragments. Moreover, T cells from C3H/HeJ spleen cell populations could be substituted for C3Heb/FeJ-derived T cells.
Monoclonal antibodies to mouse MHC antigens. II. Antibodies to the H-2Ld antigen, the products of a third polymorphic locus of the mouse major histocompatibility complex.
Authors: Ozato K , Hansen T H , Sachs D H
Published: December 2022

In order to study the fine structure of H-2L, the third H-2 locus of the mouse major histocompatibility complex, we have produced and characterized the monoclonal anti-Ld antibodies secreted by 3 different hybridoma cell lines. Two antibodies were obtained by a specific immunization directed to Ld determinants. Another antibody obtained from an immunization including, but not specific for, an L locus difference was found to detect Ld, and not Kd or Dd molecules. Although the fusions involved an H-2d myeloma variant line, no apparent adverse "autoreaction" was observed in these anti-H-2d antibody-producing hybridoma cells. All 3 antibodies exhibited cross-reactions with the antigens of certain haplotypes, and the reactivity patterns observed corresponded to the previously described alloantigenic specificities H-2.64 and H-2.65. This concordance with known H-2 specificities contrasts with the broad, unique specificity patterns exhibited by most monoclonal antibodies to H-2 antigens and may reflect a difference in the complexity of the polymorphism of the L locus relative to that of the K and D loci. Immunoprecipitation analyses using 1 of the monoclonal antibodies showed that the molecules precipitable by this reagent were indistinguishable from those precipitable by anti-Ld alloantisera. In addition, cross-reactions of Ld, Dq, and Lq molecules were revealed by sequential immunoprecipitation analyses, supporting a close evolutionary relationship of L and D loci.
Mechanism of neonatal idiotype suppression. II. Alterations in the T cell compartment suppress the maturation of B cell precursors.
Authors: Fung J , Köhler H
Published: December 2022

The cellular mechanism in neonatally suppressed BALB/c mice, which maintains the chronic suppressed state of the TEPC-15 idiotype in the antibody response to phosphorylcholine (PC), was investigated. Cells taken from these suppressed mice cannot transfer suppression to adult BALB/c or affect the in vitro response to PC of adult BALB/c spleen cells. However, spleen cells or T cells from neonatally suppressed mice given to neonatal animals induce chronic suppression of the TEPC-15 idiotype in the anti-PC response. Co-transfer of T cells from neonatally suppressed cells with normal T cells prevented the induction of suppression in neonates. Transfer of T cells from normal or keyhole limpet hemocyanin-primed BALB/c increased the expression of TEPC-15 idiotype in chronically suppressed mice, whereas T cells from neonatally suppressed were ineffective. These findings show that T cells in neonatally suppressed mice can affect the development of immature but not mature cells. The restoration of TEPC-15 expression in neonatally suppressed animals by normal T cells and the failure to induce suppression in neonates by co-transfers of T cells from normal and chronically suppressed mice demonstrate the profound role of an altered T cell compartment in sustaining chronic idiotype suppression.
Transient populations of terminal transferase positive (TdT+) cells in juvenile rats and mice.
Authors: Sasaki R , Bollum F J , Goldschneider I
Published: December 2022

Transient populations of TdT+ cells are present in peripheral blood, lung, liver, and spleen of rats and mice between birth and 7 wk of age. TdT+ blood cells appear to be post-thymic cells, as evidenced by their absence from nu/nu mice and their pattern of TdT fluorescence. TdT+ cells in lung, liver, and spleen appear to be prethymic cells. All transient populations of TdT+ cells are sensitive to dexamethasone. Only TdT+ cells in bone marrow and thymus persist into adult life.
Characterization of accessory cells required for helper T cell induction in vitro: evidence for a phagocytic, Fc-receptor, and Ia-bearing cell type.
Authors: Erb P , Stern A C , Alkan S S , Studer S , Zoumbou E , Gisler R H
Published: December 2022

The nature of the accessory cell type required for the in vitro induction of antigen-specific helper T (Th) cells to soluble antigen was determined. Two different sources of adherent accessory cells were investigated: peritoneal exudate cells (PEC) and macrophages derived from bone-marrow cells (BMM phi), which were cultured for 8 days in vitro with colony-stimulating factor. PEC and BMM phi were rosetted with EA-SRBC and separated by sedimentation into Fc gamma receptor-positive and negative fractions. Only the Fc gamma receptor-positive fraction was active in Th-cell induction. By means of the fluorescence-activated-cell sorter (FACS-II), PEC or BMM phi were separated either into phagocytic and nonphagocytic, or Ia-positive and Ia-negative cell populations. Only phagocytic and Ia-positive accessory cells induced antigen specific Th cells. In conclusion, the accessory cell is adherent, phagocytic, carries Fc gamma-receptors and Ia products, and thus expresses the most important characteristics of the macrophage lineage.
The murine Kupffer cell. II. Accessory cell function in in vitro primary antibody responses, mitogen-induced proliferation, and stimulation of mixed lymphocyte responses.
Authors: Nadler P I , Klingenstein R J , Richman L K , Ahmann G B
Published: December 2022

The ability of murine Kupffer cells to function in several in vitro immunologic systems was investigated. These cells have been shown previously to function as accessory cells in antigen-stimulated T cell proliferation in response to protein antigens. In the present study it has been demonstrated that murine Kupffer cells also are competent as accessory cells in in vitro primary antibody responses to TNP-KLH and for T cell proliferative responses to concanavalin A. In addition, murine Kupffer cells were found to be potent stimulators of mixed lymphocyte responses. These studies extend previous observations by demonstrating that Kupffer cells are competent accessory cells in several distinct in vitro correlates of in vivo immune responses. The role of Kupffer cells in in vivo immune responses, particularly those to enterically derived antigens, may require re-evaluation in the light of these findings.
Intrathymic differentiation of cytotoxic T lymphocyte (CTL) precursors. I. The CTL immunocompetence of peanut agglutinin-positive (cortical) and negative (medullary) Lyt 123 thymocytes.
Authors: Wagner H , Hardt C , Bartlett R , Röllinghoff M , Pfizenmaier K
Published: December 2022

To analyze the developmental and functional interrelationship between cortical and medullary thymocytes, the peanut agglutinin-(PNA) binding capacity was used to separate thymocytes into PNA+ (cortical) and PNA- (medullary) thymocytes. Virtually, all positively selected PNA+ thymocytes (90% of the overall thymocyte population) expressed the Lyt 123 phenotype, whereas 90% of negatively selected PNA- thymocytes expressed Lyt 1 alloantigens, about 10% being Lyt 123 thymocytes. Provided, the requirement of Lyt 1 T helper cells was bypassed by Interleukin 2, a nonspecific mediator of T help, PNA+ Lyt 123 thymocytes mounted cytotoxic T cell responses comparable in magnitude to that of peripheral T cells. Their repertoire included antigenic disparities coded for by the complete MHC complex, H-2K, I-A, H-2D, mutational events at H-2K, as well as antigenic disparities expressed on TNP conjugated- and Sendai virus-infected syngeneic cells. PNA- Lyt 123 thymocytes represent a highly reactive pool of primary cytotoxic T lymphocyte (CTL) precursors for both alloreactive and H-2-restricted CTL responses. Since PNA- thymocytes include also Lyt 1 T helper cells, PNA- responder thymocytes are able to mount autonomously (CTL responses. Our data are first to provide direct evidence that Lyt 123 cells represent a common source of alloreactive and H-2-restricted CTL precursors in unprimed lymphocyte populations. Moreover, the apparent immunocompetence of cortical PNA+ thymocytes is now explained by their lack of T helper cells.
Biochemical and biologic characterization of lymphocyte regulatory molecules. III. The isolation and phenotypic characterization of Interleukin-2 producing T cell lymphomas.
Authors: Gillis S , Scheid M , Watson J
Published: December 2022

To isolate a stable tumor cell line source of Interleukin 2 (IL-2 formerly referred to as T cell growth factor), over 40 murine leukemia and lymphoma cells as well as 9 clonal helper and killer IL-2-driven T cell lines were screened for both constitutive and mitogen-stimulated IL-2 production. A radiation-induced splenic lymphoma from the B10.BR mouse, the LBRM-33 cell line, could be stimulated to produce over 1000 units/ml of IL-2 after 24 hr exposure to T cell mitogens. Peak IL-2 activity was found in supernatants harvested from 24-hr cultures of either 1% PHA or 20 micrograms/ml Con A-stimulated LBRM-33 cells (10(6) cells/ml). IL-2 production observed in both serum-free and serum-containing cultures represented between 1000 and 5000 times the quantity of IL-2 produced in conventional cultures of mitogen-activated rat or mouse spleen cells. Peak IL-2 production by LBRM-33 cultures (stimulated at either optimal Con A or PHA concentrations or co-stimulated with suboptimal amounts of mitogen and phorbol myristate acetate) was consistently accompanied by LBRM-33 cell death. Phenotypic characterization of the producer cell revealed LBRM-33 cells to be Thy 1+, Ly 1+, Ly 2+, Ly 3+, Qa 2-3+, Qa 3.2+, Qat 4+, and Ly 5+. These studies provide further evidence that IL-2 is a T cell product and establish a source of IL-2 that will be a valuable reagent for the isolation and further molecular characterization of this immunoregulatory molecule.
Suppression by prostaglandin E1 of vascular permeability induced by vasoactive inflammatory mediators.
Authors: Fantone J C , Kunkel S L , Ward P A , Zurier R B
Published: December 2022

Systemic treatment of rats with prostaglandin E1 or with its stable derivative, 15-(S)-15-methyl-Prostaglandin E1, markedly reduce the increases in vasopermeability induced by the intradermal injection of histamine, serotonin, bradykinin, C3a, and compound 48/80. This inhibition is dependent on the dose of the vasopermeability factor and the duration of time between injection of the prostaglandin and intradermal injection of vasopermeability factor. The inhibition of associated ultrastructurally with a preservation of tight junctions between endothelial cells. The structure-function specificity of the inhibitory effects of the prostaglandin is shown by the diminished effects of prostaglandin A2, and the lack of inhibition by prostaglandin F2 alpha. These data provide evidence that PGE1 interferes with the local effects of vasopermeability mediators and may explain the earlier observations at PGE1 protects animals from immune complex-induced nephritis.
Specific cytotoxicity of a human immunoglobulin-directed Fab'-ricin A chain conjugate.
Authors: Raso V , Griffin T
Published: December 2022

The toxic A chain of ricin and the Fab' fragment of antibody directed against human immunoglobulin (Ig) have been disulfide linked via their intrinsic sulfhydryl groups. This Fab'-A chain conjugate retained the activities of its component parts. It produced effective inhibition of protein synthesis in a cellfree rabbit reticulocyte lysate system and bound tightly ti Ig determinants on human cells. Attachment of the intact molecule to the cell surface was revealed by using fluorescein-labeled antibodies directed against both its Fab' and A chain halves. The Fab'-A chain conjugate was evaluated for cytotoxicity by using human, surface Ig positive target cells and surface Ig negative control cells. In vitro toxicity was entirely selective since the conjugate produced inhibition of de novo protein synthesis, impedence of growth, as well as death and lysis, only for cells possessing surface Ig determinants. Moreover, the effects on Ig positive cells were abrogated by the addition of free human IgG that competitively blocked the Fab' combining site of this conjugate and prevented cell surface binding. Addition of lactose, which inhibits binding of whole ricin, did not influence the action of the Fab'-A chain conjugate. These results verified that the specific binding site on the antibody half of the molecule could function as a directional carrier that facilitates A chain entry into the cytoplasm. Expression of cytotoxic effects was thereby restricted exclusively to cells bearing the appropriate target site. This new conjugate molecule thus possessed both the absolute specificity of the antibody and the potent lethality of its parent toxin.
Analysis of solute diffusion across the C5b-9 membrane lesion of complement: evidence that individual C5b-9 complexes do not function as discrete, uniform pores.
Authors: Sims P J , Lauf P K
Published: December 2022

We have investigated the diffusion of radiolabeled nonelectrolytes across the membranes of resealed erythrocyte ghosts that had been treated with the terminal complement components C5b-9 and incubated under nonlytic steady state conditions. For all solutes tested, we note that diffusion across the C5b-9 lesion is retarded to rates more than 2 orders of magnitude slower than can be anticipated for a transmembrane diffusional channel of the dimensions suggested by the ultrastructure of the C5b-9 membrane lesion. Furthermore, direct measurement of the relative selectivity of the C5b-9 membrane lesions to permeation by solutes of differing molecular radii suggests that individual membrane lesions are not of uniform functional size. Data are presented that suggest that the functional heterogeneity of the membrane lesion is due to the aggregation of individual membrane bound C5b-9 complexes into larger functional units and not due to variable stoichiometry of C9 within the individual C5b-9 complex.
In vitro immune response of human peripheral lymphocytes. VI. Distribution and characterization of precursors for PHA- and protein A-induced colony-forming B cells.
Authors: Kuritani T , Kishimoto T , Muraguchi A , Yamamura Y
Published: December 2022

B cells from peripheral blood or cord blood formed colonies by stimulation with either PHA or protein A. On the other hand, tonsillar B cells did not form protein A-induced colonies, although PHA-induced colony formation was comparable to that observed in peripheral B cells. Lack of protein A-induced colony formation in tonsillar B cells was not due to the defect of helper T cells in preculture or to the presence of suppressor cells but was due to the absence of precursors for colony formation. The result showed that PHA- and protein A-induced colony-forming cells belonged to distinct subsets of B cells. Depletion of mu-bearing cells from peripheral B cells abrogated both PHA- and protein A-induced colony formation. Depletion of delta-bearing cells did not affect PHA- and protein A-induced colony formation and the population enriched with delta-bearing cells also showed colony formation. Depletion of complement receptor (CR)-positive cells removed precursors for both PHA- and protein A-induced colony formation. These results showed that precursor cells for PHA- and protein A-induced colony formation were IgM+, IgD+ and CR+ or IgM+, IgD- and CR+.
In vitro immune response of human peripheral lymphocytes. VII. Effect of anti-mu and anti-delta antibodies on B colony formation and detection of abnormal B cells in patients with juvenile rheumatoid arthritis.
Authors: Muraguchi A , Kishimoto T , Kuritani T , Yamamura Y
Published: December 2022

Effect of anti-mu and anti-delta antibodies on PHA- and protein A-induced B colony formation was studied. Anti-mu antibody at any concentrations tested did not show inhibitory or enhancing effect on colony formation. On the other hand, anti-delta antibody enhanced both PHA- and protein A-induced colony formation. Optimum concentration of anti-delta antibody for maximum enhancement was 0.1 microgram/ml. and F(ab')2 fragment of anti-delta antibody also showed comparable enhancing effect. Simultaneous addition of IgD with anti-delta antibody abrogated anti-delta-induced enhancement, and anti-delta antibody did not show any facilitation of colony formation in IgM+ IgD- cell population. In marked contrast with normal B cells, anti-mu antibody showed a remarkable enhancing effect on protein A-induced colony formation of B cells from JRA patients. F(ab')2 fragment of anti-mu antibody also showed comparable enhancing effect. Anti-mu antibody did not show any enhancement of colony formation of B cells from several other autoimmune diseases. The result indicated the presence of abnormal B cells in JRA patients.
T cell lines with dual specificity for strong Mls and H-2 determinants.
Authors: Molnar-Kimber K L , Webb S R , Sprent J , Wilson D B
Published: December 2022

To examine the relationship of T cell specificity for Mls vs H-2 determinants, BALB/c (H-2d,Mlsb)(d,b) T cells were stimulated repeatedly in vitro with H-2-compatible, Mls-incompatible DBA/2(d,a) stimulators. This line of T cells gave strong mixed-lymphocyte reactions to the priming Mlsa determinants but, in addition, gave appreciable responses to various foreign H-2 determinants. When this T cell line was subsequently stimulated over a period of 2 mo with Mlsa-negative cells of a particular foreign H-2 haplotype, e.g., H-2k, the cells gave high responses to H-2k determinants but only very low responses to third-party H-2 determinants. Significantly, the cells retained high reactivity for Mlsa determinants. In other experiments, BALB/c T cells positively selected to Mlsa,d-negative H-2-incompatible stimulator cells retained high reactivity for Mlsa determinants. The implications of these findings are discussed.
Antibody-dependent cell-mediated antibacterial activity: K lymphocytes, monocytes, and granulocytes are effective against shigella.
Authors: Lowell G H , MacDermott R P , Summers P L , Reeder A A , Bertovich M J , Formal S B
Published: December 2022

Normal human peripheral blood lymphocytes, monocytes, and granulocytes decreased the viability of Shigella flexneri in vitro in cooperation with heat-inactivated rabbit hyperimmune anti-shigella antisera. Purified K and T lymphocyte populations used before the removal of Fc receptor-bearing cells were effective in this antibody-dependent complement-free system. In contrast, B lymphocytes and T lymphocytes from which Fc receptor-bearing cells were removed exhibited no activity. In the absence of antisera, lymphocytes and monocytes were ineffective whereas granulocytes were only moderately effective even when cells from patients recently infected with shigella were used. Convalescent (but not pre-infection) sera from these patients, however, induced marked cell-mediated antibacterial activity against their homologous infecting shigella whether the cells were from uninfected or infected individuals. These data suggest that Fc receptor-bearing lymphocytes, monocytes, and granulocytes play a role in host immune defense against enteric pathogens such as shigella even in locations where complement may be functionally deficient such as among secretory mucosal surfaces.
Enhancement of the functional activities of human T cells after their interaction with SRBC.
Authors: Silva A , Lopez-Botet M , Alvarez J , de Landazuri M O
Published: December 2022

We have examined the functional consequences of the exposure of human lymphocytes to sheep red blood cells (SRBC). Peripheral blood lymphocytes (PBL) were incubated wih SRBC under optimal conditions for their interaction and, after lysis of the erythrocytes, the unfractionated PBL were examined for several T cell functions. Only after exposure to SRBC, but not after incubation with mouse, ox, chicken, or human erythrocytes, the unfractionated PBL showed an increased reactivity in the following functions: 1) Production of T cell growth factor, after PHA stimulation; 2) mitogenic response to suboptimal doses of PHA and Con A, and 3) response in mixed lymphocyte culture. Other functional activities, such as natural cytotoxicity (NK) and antibody-dependent cellular cytotoxicity (ADCC) were also enhanced by the interaction of PBL with SRBC, but the increases in cytotoxic activities were not consistently detected. Taken together, these results indicate that the interaction of PBL with SRBC has functional consequences in the reactivity of T cells producing an enhancement of several in vitro T cell functions.
Inability of mice to generate cytotoxic T lymphocytes to vesicular stomatitis virus restricted to H-2Kk or H-2Dk.
Authors: Rosenthal K L , Zinkernagel R M
Published: December 2022

H-2k mice are unable to generate cytotoxic T lymphocytes (CTL) to vesicular stomatitis virus (VSV). This apparent unresponsiveness is found for both major serotypes of VSV, VSV-Indiana and VSV-New Jersey. CTL unresponsiveness occurs despite the ability of H-2k mice to generate a humoral immune response against VSV that is comparable to that found in responder (H-2b and H-2a) strains. All H-2k mice regardless of background genes, including various Ig allotypes, were found to be nonresponders. H-2k-linked unresponsiveness mapped to both H-2Kk and H-2Dk and occurred despite the presence of responder alleles in (responder x nonresponder)F1 mice. The unresponsiveness cannot be attributed to an inability of VSV-infected H-2k target cells to express viral surface antigens of H-2 molecules. Further, unresponsiveness cannot be overcome by using secondary stimulation in vivo or in vitro. H-2k-linked unresponsiveness does not appear to be due to suppression, and no complementation has been found in various (nonresponder x nonresponder)F1 mice. Thus unresponsiveness to VSV in association with H-2Kk or H-2Dk appears to represent an extensive defect of immune responsiveness that probably occurs because VSV is not a natural mouse pathogen.
Inhibition of cytolytic T lymphocyte clones reactive with Moloney leukemia virus-associated antigens by monoclonal antibodies: a direct approach to the study of H-2 restriction.
Authors: Weiss A , MacDonald H R , Cerottini J C , Brunner K T
Published: December 2022

H-2 restriction in cytolytic T lymphocyte (CTL)-mediated lysis of syngeneic murine Moloney leukemia virus (MoLV)-induced tumor cells was studied at the clonal level by testing the inhibitory effect of monoclonal anti-H-2 antibodies on the lytic interaction between CTL clones and target cells. Large numbers of MoLV-specific CTL clones were generated by placing limiting numbers of C57BL/6 regressor (responder) spleen cells into micro-mixed leukocyte-tumor cell cultures. The clonal CTL populations thus obtained were split into 5 aliquots and tested for lytic activity in the presence (or absence) of 1 of 3 monoclonal antibodies or of an anti-whole H-2b haplotype antiserum. Two of the monoclonal antibodies were directed against H-2Db and one against H-2Kb determinants. Specificity of these reagents had been verified by demonstrating inhibition of lysis by CTL populations directed against H-2Db and H-2Kb alloantigens. In 44 of a total of 51 clones tested, results showed selective inhibition by the anti-H-2Db (and the anti-whole haplotype) reagents, and lack of inhibition by the anti-H-2Kb antibody., Of the remaining 7 clones, none was inhibited by the anti-H-2Db antibody, and 3 were inhibited by the anti-whole haplotype antiserum. These studies show that the recognition of MoLV-associated antigens by the majority of CTL clones was restricted to the H-2Db region, and that there exists limited heterogeneity in the H-2 restriction of such clones.
Human helper T cell factor(s) (ThF). I. Partial purification and characterization.
Authors: Hirano T , Teranishi T , Toba H , Sakaguchi N , Fukukawa T , Tsuyuguchi I
Published: December 2022

T cells obtained from pleural effusion of patients with tuberculous pleurisy were stimulated in vitro with PPD. The culture supernatant was shown to be substituted for T cells in PWM-induced IgG production by human B cells. As many as 0.5 to 3 x 10(9) lymphocytes were obtained from 1 patient, and about 85% of them were E-rosette positive, making it possible to purify human helper T cell factor(s) (ThF). Helper activity was sensitive to heating at 70 degrees C for 5 min and treatment with trypsin. ThF did not have conventional Ig determinants and was recovered in the 50 to 67% ammonium sulfate fraction. The purification was done by successive DEAE-Sephadex and CM-Sephadex ion-exchange chromatography, Sephadex G-100 gel filtration, and isoelectric focusing. It was shown that ThF activity was eluted by gel filtration in the fraction with a m.w. of about 20,000. Isoelectric focusing of this fraction revealed that ThF activity distributed in the pl range of 6.5 to 8.0, whereas TCGF activity was focused in a single peak at pI of 6.5, indicating the existence of a helper factor without TCGF activity.
Isolation of DNA from DNA/anti-DNA antibody immune complexes in systemic lupus erythematosus.
Authors: Sano H , Morimoto C
Published: December 2022

A low molecular weight DNA fragment was isolated from DNA/anti-DNA antibody immune complexes found in patients with active systemic lupus erythematosus (SLE). Total sera were treated with 40% saturated ammonium sulfate to isolate gamma-globulin fraction and phenolized to partition proteins and nucleic acids. After being treated with RNAase, the sample was labeled at the 5' end with 32P-phosphate and electrophoresed in an 8% polyacrylamide gel, which was dried and autoradiographed. The sample that migrated at positions of molecular weight between 20,000 and 28,000 was susceptible to DNAase I but resistant to S1-nuclease. The data suggest that the immune complexes of SLE patients contain double-stranded DNA with 30 to 40 base pairs.
IGA in human bile and liver.
Authors: Nagura H , Smith P D , Nakane P K , Brown W R
Published: December 2022

Hepatic bile IgA from 6 patients was measured by radioimmunoassay and characterized immunochemically. The concentration of IgA was 0.14 to 0.88 mg/ml. IgA associated with secretory component (SC) as well as unassociated with SC was demonstrated in all 6 samples; The proportion associated with SC (secretory IgA) was 72% to 95%. IgA and SC were localized immunocytochemically in liver and bile duct tissues by the peroxidase-labeled antibody method. Evidence favoring endocytic, SC-mediated transfer of IgA by intrahepatic and extrahepatic biliary epithelium, but not by hepatocytes, was found.
Separation and characterization of two component tumor lines within the AKR lymphoma, AKTB-1, by fluorescence-activated cell sorting and flow microfluorometry analysis. I. the coexistence of sIg+ and sIg- sublines.
Authors: Zatz M M , Mathieson B J , Kanellopoulos-Langevin C , Sharrow S O
Published: December 2022

The AKTB-1 tumor line, previously described as having both T and B cell characteristics, was separated by electronic cell sorting (ECS) on a fluorescence-activated cell sorter (FACS) on the basis of surface immunoglobulin (sIg). This resulted in 2 distinct sIg+ and sIg- subpopulations, which were transferred in graded doses (10(1) to 10(3)) into groups of normal AKR animals. The resulting splenic tumor cells were analyzed at several time points by flow microfluorometry for surface characteristics (sIg, Thy 1, Lyt 2, and Ly9) and by erythrocyte-antibody rosetting for FcR. Two distinct sublines were identifiable--one of which was sIg+, FcR-, Thy 1.1+, Lyt 2.1+, PNA+, Ly9+ and the other of which was sIg+, FcR+, Thy 1.1-, Lyt 2.1-, PNA-, Ly 9+. When these 2 sublines, generated by ECS fractionation and high dilution transfer, were serially transferred at high doses (10(5)), they maintained their unique characteristics as distinct sublines, now designated AKTB-1t and AKTB-1b.
FCS-activated blast T cells inhibit the in vitro response of memory CTL precursors to alloantigens by removal of factor(s) from MLC supernatant.
Authors: Degiovanni G , Schaaf-Lafontaine N
Published: December 2022

The inhibitory effect of spleen cells, precultured in the presence of FCS, was assayed on the memory cytolytic T lymphocytes (CTL) response to alloantigens. For this, we have used in vitro conditions in which both particulate alloantigen and MLC SN are required to allow the generation of CTL. It was shown that the CTL response was totally inhibited in the presence of 5 to 7 days precultured spleen cells. This inhibitory effect was partly due to removal, by those precultured cells, of relevant factor(s) contained in the MLC SN. After velocity sedimentation at unit gravity, it was shown that the T cells able to inhibit the cytolytic response and to remove MLC SN factor(s) are found in the fractions containing the large proliferating cells. It was further demonstrated that in the presence of inhibiting cells, a significant CTL response may be obtained after addition of concentrated MLC SN. However, in this way, this inhibitory effect was not totally circumvented, which suggests that the memory CTL response is also impaired by other mechanisms.
Induction of IgE synthesis and potentiation of anti-ovalbumin IgE antibody response by HgCl2 in the rat.
Authors: Prouvost-Danon A , Abadie A , Sapin C , Bazin H , Druet P
Published: December 2022

The present study demonstrates that mercuric chloride (HgCl2) induces a striking increase of total serum IgE in Brown Norway (BN) rats. Values up to several milligrams of IgE per milliliter of serum were observed. No antibody specificity was demonstrated for these IgE. Mercuric chloride also potentiated a specific anti-ovalbumin IgE response when the rats were immunized with ovalbumin. The kinetics of the response to HgCl2 was different from that to an antigenic stimulus alone: total IgE increased together with the potentiated anti-ovalbumin IgE antibody response to reach maximum values about 14 days after initiation of HgCl2 injections. The potentiated IgE antibodies represented only an insignificant fraction of total IgE. All these findings were observed in BN but not in Lewis rats. These data show analogies with those reported after parasitic infection in the rat and suggest similar mechanisms of action.