bluuebunny/crossref_metadata_2025 huggingface dataset:
Selective down modulation of L3T4 molecules on murine thymocytes by the tumor promoter, phorbol 12-myristate 13-acetate.
Authors: Wang P T , Bigby M , Sy M S
Published: December 2022

Treatment of murine thymocytes, but not mature peripheral T cells, with the tumor promoter, phorbol 12-myristate 13-acetate (PMA), 3 results in a rapid disappearance of L3T4 molecules from the surface of thymocytes. The effect of PMA on L3T4 molecules persists in vitro for at least 72 hr. Down modulation of L3T4 molecules was PMA dose-dependent and temperature-dependent. L3T4 molecules on cortisone-resistant thymocytes were significantly less sensitive to the effect of PMA than were L3T4 molecules on cortisone-sensitive thymocytes. Down modulation of L3T4 molecules on thymocytes did not interfere with their capacity to respond to concanavalin A or activation signals delivered via their T cell receptors. The difference in the ability of thymocytes and peripheral T cells to respond to PMA cannot be explained by differences in the number of PMA receptors. Both thymocytes and peripheral T cells have PMA receptors in the range of 1 to 1.5 X 10(5) receptors/cell. However, there is a small difference in the affinity (Kd) of the receptors on thymocytes (Kd = 30 to 40 nM) and peripheral T cells (Kd = 10 to 15 nM). Immunofluorescent staining revealed that the down modulation of L3T4 molecules by PMA was a result of internalization of L3T4 molecules. After down modulation, L3T4 could be readily detected on the cytoplasm of thymocytes. These findings suggest that L3T4 molecules on thymocytes may be subject to different regulatory signals than L3T4 molecules on peripheral T cells.
Phosphorylation of the CD8 antigen on cytotoxic human T cells in response to phorbol myristate acetate or antigen-presenting B cells.
Authors: Acres R B , Conlon P J , Mochizuki D Y , Gallis B
Published: December 2022

We have used the monoclonal antibody OKT8 to demonstrate that the cluster designation (CD)8 antigen on cytotoxic human T cells undergoes a previously unreported protein modification. Immunoprecipitation of CD8 with OKT8 from a CD8+ and CD4+ T cell line prelabeled with [32P]phosphate demonstrates that CD8 can be constitutively labeled with phosphate. CD8 undergoes rapid and intense phosphorylation in serine upon addition of phorbol myristate acetate to the cells. CD8 phosphorylation is induced upon addition of heterologous, Epstein-Barr virus-transformed B cells, which cause proliferation and are target cells for a cytotoxic CD8+CD4+ T cell line and a CD8+CD4- T cell clone. Phosphorylation induced by targets is dose-dependent, rapid, and followed by a fast dephosphorylation. Epstein-Barr virus-transformed B cells that do not induce proliferation of and are not targets for the CD8+CD4+ line and the CD8+CD4- clone do not induce CD8 phosphorylation. Cloned CD8+CD4- cells that proliferate in response to target cells, but lyse them only in the presence of lectin do not undergo target cell-induced CD8 phosphorylation. These data suggest that induction of CD8 phosphorylation is antigen-specific and is coincident with the cytotoxic response. Finally, preincubation of effector and target cells with an antibody to a monomorphic determinant of major histocompatibility complex class I antigens reduces target-induced CD8 phosphorylation to a greater extent than antibody to a major histocompatibility complex class II subregion (DR) monomorphic determinant, reinforcing the notion that major histocompatibility complex class I antigens interact with CD8+ cells.
Rejection of fetal neocortical neural transplants by H-2 incompatible mice.
Authors: Nicholas M K , Antel J P , Stefansson K , Arnason B G
Published: December 2022

In order to examine questions concerning immunologic privilege of the central nervous system, we placed neocortical transplants into cerebral ventricles of mice. We compared the fates of transplants between fully H-2 compatible (isografts) and H-2 incompatible (allografts) animals. Histologic evaluation comparing animals from iso- and allograft groups revealed significant differences in the number of inflammatory cells and in the degree of necrosis within the grafts. Response to allografted tissue within the brain mimics that seen in several immune-mediated diseases of the nervous system in that neurons appear to be selectively spared. Only upon subsequent stimulation of the host's immune system with an orthotopic skin graft bearing the major histocompatibility complex antigens of the neural graft are neurons destroyed. Immunohistochemical evaluation revealed that the inflammatory cell infiltrates in and around the allografts were composed of Lyt-2+, L3T4+, and Mac-1+ cells. In addition, Ia+ endothelial cells as well as Ia+ parenchymal CNS cells were found in both donor and host tissue of allografted animals. Hence, H-2 incompatible neural tissue transplanted to the CNS is recognized and rejected by the immune system of the recipient animal. The cellular infiltrates seen within the first weeks to months following transplantation of allogeneic CNS tissue resemble those seen in other allografts undergoing rejection. We conclude that the CNS is not unconditionally privileged as either a transplant site or as a source of transplanted tissue.
Effect of B cell stimulatory factor-1 (interleukin 4) on Fc epsilon and Fc gamma receptor expression on murine B lymphocytes and B cell lines.
Authors: Conrad D H , Waldschmidt T J , Lee W T , Rao M , Keegan A D , Noelle R J , Lynch R G , Kehry M R
Published: December 2022

Culture of murine splenic B cells with interleukin 4 (IL-4) caused the up-regulation of the lymphocyte Fc receptor for immunoglobulin E (IgE) (Fc epsilon R) over a similar dose range as required for Ia up-regulation. However, the expression level of the Fc receptor for immunoglobulin G (Fc gamma R) did not increase, rather IL-4 caused a slight but consistent decrease in the Fc gamma R level on the B cells. Fc epsilon R+ B hybridoma cells also responded to IL-4 by exhibiting increased Fc epsilon R expression; with the hybridoma cells Fc gamma R levels were unaffected. IL-4 caused an increase in the number of Fc epsilon R per cell and the highest levels of expression were obtained by having both IgE and IL-4 present in the culture. The specificity of the increase was demonstrated by blocking IL-4-mediated actions with monoclonal anti-IL-4 (11B11). Experiments following the incorporation of [35S]methionine into the Fc epsilon R demonstrated that IL-4 increased the rate of Fc epsilon R biosynthesis; this provides an explanation for the IL-4-induced increase in Fc epsilon R expression. IL-4, unlike IgE, had no effect on the rate of degradation of the Fc epsilon R. Interferon-gamma (IFN-gamma) totally abrogated IL-4-mediated Fc epsilon R up-regulation; at the same concentration of IFN-gamma Ia up-regulation is also suppressed, although not as effectively. IFN-gamma was shown to directly suppress Fc epsilon R synthesis, thereby explaining the inhibitory action on Fc epsilon R levels. Finally, it was shown that 11B11 inhibited the increased expression of Fc epsilon R on B cells obtained from mice during the early, but not the late, stages of Nippostrongylus brasiliensis infection. This latter finding suggests that the high Fc epsilon R levels seen early in parasite infections are dependent upon IL-4. The results overall provide further insight into the biologic activities of IL-4.
Identification of variants of the basophilic leukemia (RBL-2H3) cells that have defective phosphoinositide responses to antigen and stimulants of guanosine 5'-triphosphate-regulatory proteins.
Authors: Woldemussie E , Ali H , Takaishi T , Siraganian R P , Beaven M A
Published: December 2022

Antigen immunoglobulin E-mediated secretion of histamine from RBL-2H3 cells is associated with substantial hydrolysis of membrane inositol phospholipids and a rise in the concentration of cytosol Ca2+ (calcium signal). Such responses differed among cloned variant lines of the RBL-2H3 cell line from undetectable (1A3 bromodeoxyuridine-resistant (BUDRR), 2B1 BUDRR, and 1B3 BUDRR lines) to about 80% of those in the parent RBL-2H3 cells. In all but one line (1B3 thioguanine-resistant (TgR)), the intensities of the phosphoinositide response and of the calcium signal were correlated with the secretory response. The 1B3 TgR line had no detectable calcium signal (as measured by quin 2 fluorescence or uptake of 45Ca2+) but paradoxically showed modest rates of hydrolysis of inositol phospholipids and of secretion. The responses of the 1B3 TgR line were, however, dependent on the presence of external Ca2+ ions. The induction of secretion with antigen, therefore, was invariably associated with the hydrolysis of inositol phospholipids, but it was not necessarily associated with a change in concentration of cytosol Ca2+. All antigen unresponsive clones could secrete when synergistic signals were induced by exposure to the Ca2+ -ionophore, A23187 and the phorbol ester, 12-O-tetradecanoylphorbol 13-acetate. These lines, otherwise, had immunoglobulin E receptors and had no obvious defect in their capacity to synthesize the inositol phospholipids or in their phenotypic expression of phospholipase C as measured in cell extracts. One finding of possible relevance to the role of guanosine 5'-triphosphate-regulatory proteins in the activation of phospholipase C was the inability of one antigen-nonresponsive line to respond to NaF (in intact cells) or to guanosine 5'-(3-O-thio)triphosphate (in electrically permeabilized cells).
Glucocorticoids suppress calcium mobilization and phospholipid hydrolysis in anti-Ig antibody-stimulated B cells.
Authors: Dennis G , June C H , Mizuguchi J , Ohara J , Witherspoon K , Finkelman F D , McMillan V , Mond J J
Published: December 2022

Glucocorticoids have been shown to play a major role in influencing the activation of B lymphocytes. In view of our recent observation that dexamethasone exerts a marked suppressive effect on an early event in B cell activation that is stimulated by anti-Ig antibody, we investigated its activity on other stimuli that induce intracellular events similar to those produced by anti-Ig antibody. Because the intracellular events that occur after B cell stimulation with phorbol myristate acetate and the calcium ionophore A23187 appear to mimic those that occur after B cell stimulation with anti-Ig antibody, we studied whether the cellular responses elicited by these activation stimuli are affected in a similar fashion by dexamethasone. Whereas anti-Ig antibody-stimulated entry of G0 B cells to the G1 and S phase of the cell cycle was markedly suppressed by dexamethasone, phorbol myristate acetate/A23187 stimulation of these events was resistant to dexamethasone. Our finding that anti-Ig-induced cross-linking of B cell surface Ig, as measured by surface Ig capping, was not inhibited by dexamethasone suggested that corticosteroids inhibit anti-Ig-induced B cell proliferation at a step distal to membrane Ig cross-linking and proximal to phosphatidylinositol bisphosphate hydrolysis. This hypothesis is supported by experiments presented in this manuscript which demonstrate that dexamethasone inhibits anti-Ig-stimulated phosphatidylinositol bisphosphate hydrolysis. We also found that dexamethasone markedly inhibited anti-Ig antibody-stimulated increases in intracellular ionized calcium concentrations. This dexamethasone-mediated suppression is time-dependent as it is not seen when B cells are cultured with dexamethasone for less than 6 hr. Our data suggest that the immunomodulatory activity of glucocorticoids is exerted by binding to its nuclear receptor, thereby preventing the generation of second messengers required for cell activation after agonist-receptor interaction.
Regulation of murine IgE production in SJA/9 and nude mice. Potentiation of IgE production by recombinant interleukin 4.
Authors: Azuma M , Hirano T , Miyajima H , Watanabe N , Yagita H , Enomoto S , Furusawa S , Ovary Z , Kinashi T , Honjo T
Published: December 2022

Serum IgE levels were determined in different strains of mice with enzyme-linked immunosorbent assay by using rat monoclonal anti-murine IgE antibodies in normal and in Nippostrongylus brasiliensis-infected mice. After infection, serum IgE levels were high in BALB/c and CB-20, low in SJL/J and SJA/20 mice, and not detected at all in SJA/9 and nude mice. Surface IgE-positive cells were greatly increased in BALB/c and SJL/J mice after infection, but not in SJA/9 and nude mice. Most surface IgE-positive spleen cells were also surface IgM- and surface IgD-positive. When spleen cells from SJA/9 or nude mice were stimulated in vitro with lipopolysaccharide and recombinant interleukin 4 (formerly B cell-stimulating factor 1), IgE was produced and detected in the supernatants of these cultures. In addition, surface IgE-positive cells could be detected in these cultures. Most of the surface IgE-positive cells were surface IgM- and surface IgD-negative, unlike those seen in the spleens of Nippostrongylus-infected BALB/c and SJL/J mice. These observations show that SJA/9 and nude mice have IgE-producing precursor B cells, and after appropriate stimulation interleukin 4 can induce them to secrete IgE.
Antigen presentation by hapten-specific B lymphocytes. V. Requirements for activation of antigen-presenting B cells.
Authors: Liano D , Abbas A K
Published: December 2022

Splenic B cells specific for the haptens, 2,4,6-trinitrophenyl (TNP) or fluorescein isothiocyanate (FITC) were cultured with a range of concentrations of unmodified or TNP- or FITC-conjugated conalbumin and the conalbumin + I-Ak-specific, interleukin (IL) 1-dependent helper T cell clone, D10 . G4, in the presence and absence of IL-1. Lymphokine secretion, T cell proliferation, and antibody secretion by B cells all exhibited identical antigen dose responses. Thus, hapten-binding B cells presented low concentrations of haptenated conalbumin for activation of both the T and the antigen-presenting B cells. Whereas proliferation of D10 . G4 required the addition of IL-1, both lymphokine production and stimulation of B cells to antibody secretion occurred without exogenous IL-1. These results demonstrate that when B lymphocytes function as presenting cells for antigens that bind to their immunoglobulin receptors, activation of the responding T cells and the B cells themselves occur at similar concentrations of antigen. Moreover, for functional T-B interactions, antigen-presenting B and responding T lymphocytes constitute a complete system that requires no other accessory stimuli, whereas clonal expansion of T cells is dependent on accessory factors such as IL-1. Finally, since D10 . G4 secretes IL-4 but neither IL-2 nor interferon-gamma, our results demonstrate that differentiation of B cells as a consequence of direct ("cognate") interactions with helper T cells as well as of bystander B cells can occur in the absence of IL-1, IL-2, and interferon-gamma.
Production of cytotoxic factor for oligodendrocytes by stimulated astrocytes.
Authors: Robbins D S , Shirazi Y , Drysdale B E , Lieberman A , Shin H S , Shin M L
Published: December 2022

Stimulation of rat astrocytes in vitro by calcium ionophore A23187 and/or lipopolysaccharide results in the generation of a cytotoxic factor that is functionally similar to the previously described macrophage-derived cytotoxic factor, tumor necrosis factor. Like the macrophage product, the astrocyte cytotoxic factor kills murine L 929 cell targets. In addition, it kills rat oligodendrocytes, the myelin-producing cells of the central nervous system. Human recombinant tumor necrosis factor also has cytotoxic activity directed against rat oligodendrocytes.
Activation of plasma Hageman factor and kallikrein in ongoing allergic reactions in the skin.
Authors: Atkins P C , Miragliotta G , Talbot S F , Zweiman B , Kaplan A P
Published: December 2022

Ten atopic subjects, sensitive to intradermal injection of less than or equal to 10 protein nitrogen units of ragweed or grass pollen antigen, underwent paired antigen and buffer skin chamber incubation over the base of denuded skin blisters. The chamber fluids were sampled over a 6-hr period for histamine and activated Hageman factor and plasma kallikrein which were complexed to C1 inhibitor. In 9 of 10 subjects significantly (p less than 0.01) increased histamine levels (74 +/- 11 ng/ml vs 1.5 +/- 0.55 ng/ml) and kallikrein-C1 inhibitor complexes (2.15 +/- 0.78 ng/ml/hr vs 0.51 +/- 0.09 ng/ml/hr, p less than 0.25) were detected at antigen sites compared with buffer sites, respectively. Increased levels of activated Hageman factor (ng/ml/hr) were detected at antigen sites (1.35 +/- 0.60) compared with buffer sites (0.11 +/- 0.05), (p less than 0.01), in 8 of 10 subjects. Whereas peak levels of histamine were obtained after 1 hr of challenge, both Hageman factor and kallikrein activation, as assessed by complex formation, tended to peak later from the 2nd to the 5th hr. This represents the first demonstration that cutaneous IgE-mediated allergic responses are associated with local activation of the intrinsic plasma coagulation-kinin pathways.
Monoclonal antibody characterization of Plasmodium falciparum antigens in immune complexes formed when schizonts rupture in the presence of immune serum.
Authors: Chulay J D , Lyon J A , Haynes J D , Meierovics A I , Atkinson C T , Aikawa M
Published: December 2022

When Plasmodium falciparum parasites are cultured with some immune sera, merozoites are agglutinated by antibodies to form immune clusters of merozoites and prevent their invasion into erythrocytes. Within these immune clusters of merozoites, several antigens that are normally found in the soluble fraction after detergent extraction accumulate in relatively insoluble immune complexes. From mice immunized with these immune complexes, we obtained hybridomas secreting monoclonal antibodies (mAb) that react with various immune clusters of merozoites antigens, including mAb 3D5, which recognizes a 101-kDa antigen (p101) and mAb, 5E3, which recognizes a 113-kDa antigen (p113). Both mAb reacted with antigens at the surface of schizonts, in the vacuolar space, and at the surface of merozoites before their release from schizont-infected cells. Both p101 and p113 were synthesized by mature trophozoites and young schizonts. In pulse-chase experiments, p113 was processed to 100-, 70-, 55-, and 50-kDa products. Both p101 and p113 appeared in the culture medium when schizont rupture occurred in normal culture medium but were found in immune complexes when schizont rupture occurred in the presence of immune serum. Antibodies in immune complexes, when dissociated with acid and used to probe immunoblots, reacted with affinity-purified p101 and p113. Antigens such as these, which are accessible at the parasite surface and react with antibodies present in immune serum that inhibits parasite invasion, are logical candidates to study in the search for a vaccine against the erythrocytic stages of malaria.
IgG antibody subclasses in human filariasis. Differential subclass recognition of parasite antigens correlates with different clinical manifestations of infection.
Authors: Hussain R , Grögl M , Ottesen E A
Published: December 2022

The four subclasses of IgG are distinct in structure, function, and degree of participation in the antibody response to complex antigens. Looking for differential responsiveness of potential pathogenetic significance, we have analyzed both quantitatively and qualitatively the filaria-specific IgG subclass responses of 20 patients with lymphatic filariasis presenting either with chronic lymphatic obstructive pathology and elephantiasis (CP) or with asymptomatic microfilaremia (MF). Subclass-specific monoclonal antibodies were used in an enzyme-linked immunosorbent assay to study IgG filarial antibodies quantitatively and in immunoblot analyses to determine qualitatively the subclass antibody specificities. Quantitatively, the most significant differences among patient groups were in levels of IgG4, which were more than 17 times higher in MF patients (geometric mean, 64.7 micrograms/ml) than in those with CP (mean, 3.7 micrograms/ml). When qualitative analyses were done on the same sera, major differences were noted, particularly in the recognition profiles of the IgG1, IgG3, and IgG4 responses. IgG1 and IgG3 responses to antigens were seen especially to antigens with m.w. greater than 68,000 in all patients with elephantiasis, whereas MF patients showed most of their reactivity to antigens less than 68,000. For IgG4, the MF patients showed prominent recognition of antigens throughout the entire range of m.w., whereas those with CP had very little IgG4 recognition of antigens of any m.w. Interestingly, this relationship was essentially reversed in the IgG3 antibody responses (especially to antigens greater than 68,000) and, to a lesser extent, the IgG1 responses. These findings demonstrate correlations of potential cause/effect significance between IgG4 antibody responsiveness and the immunomodulated asymptomatic MF form of clinical filariasis and between IgG3/IgG1 antibody responsiveness and the clinical presentation of CP.
Regulation by anti-CD2 monoclonal antibody of the activation of a human T cell clone induced by anti-CD3 or anti-T cell receptor antibodies.
Authors: Yssel H , Aubry J P , de Waal Malefijt R , de Vries J E , Spits H
Published: December 2022

In this study the effect of anti-cluster designation (CD) 2 monoclonal antibodies (mAb) on the activation of a cloned human T cell line, HY837, after triggering the CD3/T cell receptor (TcR) complex by anti-CD3 or anti-TcR mAb is described. HY837, which reacts with a series of mAb directed at different epitopes on the TcR, could be induced to proliferation and interleukin 2 (IL-2) production by soluble mAb directed at the CD3/TcR complex in the absence of accessory cells. mAb directed at the CD2 epitope T11-1 were shown to block the IL-2 production by HY837, as well as the expression of the IL-2 receptor, induced by anti-CD3 mAb, resulting in the inhibition of the proliferative response. The effect of anti-CD2 mAb on the proliferative response of HY837, induced by anti-CD3 mAb, was not due to a competition for Fc binding sites. In contrast, the proliferative responses and IL-2 production of HY837, induced by mAb directed at the TcR, were shown to be enhanced by the action of the anti-CD2 mAb. These results indicate that effects mediated by anti-CD3/TcR mAb cannot always be extrapolated to antigen-mediated effects and show that anti-CD2 mAb may regulate the T cell response, induced by mAb directed at the CD3/TcR complex, depending on which part of this complex is triggered during activation.
Immune response to human immunodeficiency virus. In vivo administration of anti-idiotype induces an anti-gp160 response specific for a synthetic peptide.
Authors: Zhou E M , Chanh T C , Dreesman G R , Kanda P , Kennedy R C
Published: December 2022

Anti-idiotypic antibodies (anti-Id) to chimpanzee antibodies directed against a synthetic peptide corresponding to a native epitope associated with gp41 of human immunodeficiency virus (HIV) envelope glycoprotein were produced in rabbits. The peptide was analogous to amino acid sequences 735 to 752 from the human T cell leukemia virus-IIIB isolate of HIV. Characteristics of the anti-Id preparation included: 1) detection of a shared determinant present on a second chimpanzee and one of three rabbit antibody preparations directed against the synthetic peptide, 2) failure to recognize an idiotype (Id) in BALB/c mouse antisera to the peptide, and 3) partial inhibition of the homologous chimpanzee Id preparation from binding either peptide or a recombinant HIV gp160 preparation. Immunization of BALB/c mice with the anti-Id induced an antipeptide response which bound a recombinant gp160 preparation without subsequent peptide or gp160 exposure. The anti-gp160 containing sera from mice immunized with anti-Id were able to inhibit the Id-anti-Id reaction indicating that an Id-positive antibody response was induced. This Id is not normally expressed in the murine anti-gp 160 immune response to the synthetic peptide and suggests that this anti-Id may activate normally silent clones. This study indicates that Id networks may be operational during the immune response to HIV epitopes. Alternatively, anti-Id may be useful in altering the serologic characteristics of an antibody response to HIV and may offer potential for modulating the immune response in this viral infection.
Interleukin secretion by B cell lines and splenic B cells stimulated with calcium ionophore and phorbol ester.
Authors: Taira S , Matsui M , Hayakawa K , Yokoyama T , Nariuchi H
Published: December 2022

A B cell lymphoma A20.2J and splenic B cells produced an active material to support the proliferation of an interleukin 2 (IL-2)-dependent T cell line, CTLL-2, by stimulation with both calcium ionophore A23187 and phorbol myristate acetate (PMA). Although the production of the active material was induced by stimulation with A23187 alone in A20.2J cells, both A23187 and PMA were essential for the stimulation of splenic B cells. Neither A20.2J cells nor splenic B cells produced the active material by stimulation with PMA alone. The production was inversely proportional to the concentration of fetal calf serum in culture medium. The active material produced by B cells was indicated to be IL-2 and not B cell-stimulating factor 1 (BSF-1) for the following reasons: 1) the proliferation of CTLL-2 cells in the presence of active material was inhibited by the inclusion of anti-IL-2 receptor or anti-IL-2 in culture medium but not by anti-BSF-1; 2) the material showed no co-mitogenic activity to purified splenic B cells with anti-immunoglobulins and did not support the proliferation of FDC-P2 which are known to grow in the presence of BSF-1; and 3) IL-2 mRNA could be detected in A20.2J and splenic B cells stimulated with A23187 and PMA in Northern blot analysis. Some B cell hybridomas were also shown to produce IL-2 by similar stimulation to A20.2J. Splenic B cells as well as A20.2J cells were able to produce IL-2 by stimulation with anti-immunoglobulins. These results suggest that under certain conditions IL-2 can be produced by splenic B cells, at least some subsets of B cells, and B cell lines.
Expression of tumor necrosis factor receptors on human monocytes and internalization of receptor bound ligand.
Authors: Imamura K , Spriggs D , Kufe D
Published: December 2022

Tumor necrosis factor (TNF) is a polypeptide produced by monocytes and macrophages. Although TNF receptors have been identified on a variety of cell types, previous studies have not determined whether these receptors also exist on monocytes. In the present work, highly purified recombinant TNF was labeled with 125I. The 125I-labeled TNF bound specifically to receptors on human monocytes and monocyte membrane preparations. A curvilinear Scatchard plot indicated the presence of TNF-binding sites with two different affinities. The results also indicate that receptor-bound TNF is rapidly internalized by monocytes and then degraded intracellularly. These findings are in concert with recent studies demonstrating that TNF immunomodulates monocyte function by an autocrine mechanism.
Inhibition of human neutrophil chemotaxis by the protein kinase inhibitor, 1-(5-isoquinolinesulfonyl) piperazine.
Authors: Harvath L , McCall C E , Bass D A , McPhail L C
Published: December 2022

The protein kinase inhibitor, 1-(5-isoquinolinesulfonyl) piperazine (C-I), inhibits superoxide release from human neutrophils (PMN) stimulated with phorbol myristate acetate or synthetic diacylglycerol, without inhibiting superoxide release from PMN stimulated with the chemoattractants C5a or N-formyl-methionyl-leucyl-phenylalanine (f-Met-Leu-Phe). In this study, we investigated the effect of C-I on human PMN chemotaxis to C5a, f-Met-Leu-Phe, leukotriene B4 (LTB4), and fluoresceinated N-formyl-methionyl-leucyl-phenylalanine-lysine (f-Met-Leu-Phe-Lys-FITC). PMN, preincubated for 5 min at 37 degrees C with 0 to 200 microM C-I, were tested for their migratory responses to the chemoattractants. C-I (greater than or equal to 1 microM) significantly inhibited PMN chemotaxis to f-Met-Leu-Phe, f-Met-Leu-Phe-Lys-FITC, and C5a without affecting random migration. Maximal inhibition of chemotaxis to these attractants occurred with greater than or equal to 50 microM C-I, at which chemotaxis was inhibited by 80 to 95%. The C-I inhibition was reversible. In contrast, 200 microM C-I did not inhibit the number of PMN migrating to LTB4, although, the leading front of PMN migration to LTB4 was inhibited by C-I. C-I inhibited PMN orientation to C5a and f-Met-Leu-Phe without affecting orientation to LTB4. C-I did not inhibit the binding of radiolabeled f-Met-Leu-Phe or f-Met-Leu-Phe-Lys-FITC to PMN. These findings suggest that the chemotactic responses of PMN to f-Met-Leu-Phe and C5a involve a protein kinase-dependent reaction which is inhibited by C-I.
Sequence and type-specific immunogenicity of the amino-terminal region of type 1 streptococcal M protein.
Authors: Kraus W , Haanes-Fritz E , Cleary P P , Seyer J M , Dale J B , Beachey E H
Published: December 2022

The NH2-terminal sequence of type 1 M protein was determined by automated Edman degradation of purified polypeptide fragments extracted from whole streptococci by limited digestion with pepsin. Three polypeptide fragments were purified by slab gel electrophoresis on sodium dodecyl sulfate (SDS) polyacrylamide followed by electroelution. The purified fragments migrated as 28-, 25-, and 23.5-kDa fragments, respectively. Each of the fragments inhibited opsonization of a diluted antiserum prepared in rabbits by immunization with whole type 1 streptococci. The amino-terminal sequences of the peptide fragments were confirmed by comparison with the primary structure predicted from the nucleotide sequence of the type 1 M protein structural gene. The 28-kDa fragment contained the NH2-terminal asparagine residue of the processed type 1 M protein, whereas the NH2-terminal sequences of the 25- and 23.5-kDa peptides began at residues 27 and 36, respectively. A seven-residue periodicity with respect to polar and nonpolar residues was observed beginning at residue 22 and, therefore, the secondary structural potential of type 1 M protein is similar to that reported for other M proteins. In contrast to the other M proteins, however, identical repeats were rare, the longest sequence identity consisting of a three-amino acid acid sequence Lys-Asp-Leu at positions 30-32 repeated once at positions 65-67. A 23-residue synthetic peptide of the amino-terminus of the type 1 M protein evoked opsonic antibodies against type 1 streptococci. These results indicate that the NH2-terminal region of type 1 M protein retains the secondary structural characteristics of other M serotypes. Moreover, it contains epitopes that evoke protective immune responses. Our studies may have bearing in the development of safe and effective vaccines against group A streptococcal infections.
Phenylhydrazine stimulates lymphopoiesis and accelerates Abelson murine leukemia virus-induced pre-B cell lymphomas.
Authors: Klinken S P , Holmes K L , Fredrickson T N , Erner S M , Morse H C
Published: December 2022

Infection of bone marrow or fetal liver cells with Abelson murine leukemia virus (A-MuLV) results in the transformation of pre-B cells and the development of erythroid colonies, indicating that the abl oncogene can affect the growth characteristics of immature cells in both the B cell and erythroid lineages. By comparison, infection of mice with A-MuLV results primarily in the development of pre-B cell lymphomas. To determine whether A-MuLV could induce erythroid disease in vivo, NFS/N mice were pretreated with phenylhydrazine (PHZ) to stimulate erythropoiesis and increase the frequency of potential target cells for A-MuLV. No erythroleukemias developed in mice treated with PHZ. Instead, the latency for pre-B cell lymphomas was reduced by half. This acceleration of disease could be attributed to a marked increase in pre-B cells as targets for transformation by A-MuLV in the bone marrows but not the spleens of treated mice. Increases in the frequencies of T cells in bone marrow and spleen also followed treatment with PHZ. These results show that although PHZ-induced anemia stimulates the production of T and B cells as well as erythroid progenitors, PHZ-treated mice do not develop erythroleukemia or T cell lymphomas. It was also found that the genetically determined resistance of adult C57BL/6 mice to lymphoma induction by A-MuLV could not be overcome by pretreatment with PHZ even though the frequency of pre-B cells in bone marrow was greatly increased by this treatment.
Vaccination against cutaneous leishmaniasis in a murine model. II. Immunologic properties of protective and nonprotective subfractions of soluble promastigote extract.
Authors: Scott P , Pearce E , Natovitz P , Sher A
Published: December 2022

We have previously demonstrated that BALB/c mice can be protected against a fatal infection with Leishmania major by i.p. immunization with a soluble leishmanial antigen (SLA) preparation in conjunction with the adjuvant, Corynebacterium parvum (CP). In this study, SLA was separated into nine distinct fractions by anion exchange liquid chromatography, and the fractions were analyzed for their ability to stimulate T cells obtained from immunized mice, to be recognized by vaccine-induced antibodies, and to induce protective immunity. While all but one of the fractions were recognized by antibodies from SLA + CP immunized mice, only two fractions (fractions 1 and 9) stimulated lymphocytes to produce macrophage-activating factor and elicited significant delayed-type hypersensitivity in vivo. When mice were immunized with the fractions, only fraction 9 stimulated significant immunity (76% protection in seven experiments). Proteins (accounting for 1.3% of the total in SLA) appear to be responsible for the protection elicited with fraction 9, since protease treatment of this fraction destroyed its immunogenicity. Thus, a partially purified protective protein antigen fraction has been obtained and protection with this fraction correlated with cell-mediated immune responses. However, these results also demonstrate that the ability of leishmanial antigens to be recognized by T cells and produce macrophage-activating factor does not in itself predict whether such molecules will induce immunity, suggesting that protective leishmanial antigens may have additional unique properties.
A Colorimetric Reaction Between Gold Chloride and Toxins Apparently Indicative of Toxic Strength
Authors: Mishulow Lucy , Krumwiede Charles
Published: December 2022

By means of a colorimetric reaction obtained by adding gold chloride to toxic-broth filtrates we have been able to obtain toxin values which in most instances closely approximate the toxin values obtained by animal tests. Thus far we have not observed a colorimetric end-point which parallels the combining value (toxin-toxoid) of toxins.

We have no data as to the mechanism of the reaction. Whether the reaction is due to a specific toxin-gold chloride combination, or whether some other factor or factors are involved remains to be proven.

Whatever the mechanism of the reaction may be, the results strongly suggest that a practical, gold chloride colorimetric method of standardizing toxic-values of broth filtrates may be developed.
On the Heterogenetic Haptene
Authors: Landsteiner K. , Levene P. A.
Published: December 2022

In attempting to purify the specific part of the heterogenetic haptene, present in alcoholic extracts of horse kidneys, fractions were obtained which are distinguished from the common lipoids by their solubility in water. The material of one fraction described recently3 is practically insoluble in alcohol, chloroform and ether and gives a viscous solution in hot water. With orcinol and copper it gives no purple color reaction in contrast to other water-soluble fractions; it contains neither phosphorous nor sulfur.

It was possible to obtain from this serologically active product two fractions differing in their composition. The method of preparation was the following. The crude material separating from alcoholic extracts of horse kidneys and freed from ether soluble substances1 was dissolved in 20 parts of hot pyridine and allowed to stand in the ice box for forty-eight hours. After removing the sediment the filtrate was evaporated under reduced pressure to nearly dryness.
Distribution of Immune Agglutinins in the Serum and Organs of Rabbits
Authors: Freund Jules
Published: December 2022

The purpose of the present study was to ascertain by a direct method the eventual accumulation of antibodies in the organs of rabbits which had received injections of rabbit blood containing agglutinins. The presence of antibodies in the tissues of animals injected with homologous serum containing anaphylactic antibodies or precipitins seemed probable, (1) as an explanation of the incubation period of anaphylaxis, (2) on the ground of the observation that anaphylaxis once developed is independent of the circulating antibodies. Furthermore, it was assumed that during the time of contact of antibodies with the tissue, the relation of the two changes; after the antibodies have been in contact with the tissue for a period, they become fixed to the tissue.1 We have attempted to obtain direct evidence for this assumption by means of perfusion experiments.

In their investigation concerning the site of antibody formation, Pfeiffer and Marx (1), as early as 1898, demonstrated by recovery the presence of antibodies in the spleen.
Cell Penetration by Venom Hemotoxin
Authors: Kyes Preston , Bailey John Hays
Published: December 2022

Flexner and Noguchi in 1902 (1) pointed out that washed erythrocytes which are not hemolyzed by venom alone, do hemolyze upon the addition of a complementing serum. They stated at that time, that no erythrocytes, regardless of species, are dissolved by venom alone. Kyes (2) later in the same year, however, showed that the erythrocytes of many species are dissolved by venom alone and that the necessity of an activating serum obtains only in a restricted number of species (sheep, goat and beef) and the same author has more recently found that even the corpuscles of the latter species are dissolved by Cobra venom alone when the corpuscles are washed and suspended in an isotonic sugar solution instead of physiological salt solution.

At present, therefore, the venom hemotoxins instead of being considered incompetent in producing hemolysis unaided by complements, must be recognized as highly efficient producers of hemolysis even in the absence of added activating substances.
Observations on Irregularities in Human Isohemagglutinins
Authors: Lacy G. R.
Published: December 2022

Four patients, acutely sick, are reported whose blood sera contained abnormal agglutinating substances for all four groups of human red corpuscles. One patient's blood contained autoisoagglutinins, while the other three did not. Three of the patients had bacteremias due to encapsulated organisms. The disturbance of the isoagglutinins was not constant, but in one case ran practically parallel with the temperature curve.

Experimental evidence is offered to show that the disturbance of the isoagglutinins was probably not due to a change in pH of the blood nor to the hyperpyrexia per se. Other evidence is presented showing that there are members of the encapsulated, gram-negative group of bacteria, namely, certain strains of B. acidi lactici and B. lactis aerogenes, together with their products, which are capable of causing agglutination of human red corpuscles.

Emphasis is here laid upon the importance of invariably crosstesting serum and corpuscles of donor and recipient, before transfusion, even though they are members of the same isoagglutinin group.
A Note on Anaphylactic Behavior in the Paramoecium
Authors: Ramsdell Susan Griffith
Published: December 2022

Paramoecia were sensitized by treatment with an anti-egg-white serum, and the response on treatment with egg-white was shown to be analogous to that in the sensitized smooth muscle. Likewise, the reaction to dilute barium chloride solution was strikingly similar to the anaphylactic response, suggesting the contractile substance as the mechanism of the reaction.
Investigation of the Cataphoresis of Toxin and Antitoxin in the “Field and Teague” and in Other Apparatus
Authors: Maver Mary E. , Falk I. S.
Published: December 2022

Numerous investigations of the nature of toxin and antitoxin have supported the view that they are amphoteric colloids, and either are proteins, or are closely adherent to proteins (1). Field and Teague (2) in 1907, however, published, the results of an experiment which indicated that both toxin and antitoxin were positive colloids which migrated to the cathode whether the solution containing them was acid or alkaline. This work, if confirmed, would afford a means for the separation of toxin and antitoxin from protein, since protein in alkaline solution is known to migrate to the anode. Because the exact hydrogen ion concentrations of the solutions used were not determined by these authors, we decided to electrolyze toxin and antitoxin solutions at a series of definite hydrogen ion concentrations, using the apparatus which they described.
A Study of Diphtheria Antitoxic Serum of Exceptionally High Potency with a Comparative Analysis of its Various Constituents
Authors: Kirkbride Mary B. , Murdick Philip P.
Published: December 2022

Horses vary markedly in their response to immunization. The reasons for this divergence even under identical conditions of treatment are still obscure despite attempts made by various investigators to correlate the phenomenon with measurable factors. An unusually favorable opportunity was afforded to ascertain by comparative tests whether changes in organic or inorganic constituents of the blood accompanied the development of antitoxin, when the serum of a horse undergoing active immunization against diphtheria toxin reached a titer of 2200 units per cubic centimeter. Since such a high titer appears to be almost unprecedented and the long period during which the horse continued to produce serum of high antitoxic potency is unusual, it has seemed of interest to include a short history of the animal with the results of the analysis of its serum in comparison with other sera.

Horse 122, a gelding, twelve years old and weighing about 900 pounds, was received from the New York State Department of Police.
Further Observations on the Effect of Type-I Pneumococcus Culture Broth on the Phagocytic Action of Type-I Pneumococcus Serum
Authors: Sickles G. M.
Published: December 2022

The inhibitory action of pneumococcus broth on phagocytosis in the previous experiments, as in these, appeared to be the result of reactions between the culture broth and the immune serum. In the experiments recorded in this paper, these reactions between the immune serum and culture broth paralleled and appeared to be associated with the reaction by which the precipitative activities of the serum were absorbed when treated with the homologous culture broth.
Effect of Dehepatization on Erythrocyte Anaphylaxis in Dogs
Authors: Boone T. H. , Chase E. M.
Published: December 2022

In a previous paper (1) it was shown that the latent period between the intravenous injection of antigen into hypersensitive dogs and the resulting precipitous fall in arterial blood pressure is much shorter in erythrocyte anaphylaxis than in serum anaphylaxis. In serum anaphylaxis this latent period is usually about forty-five seconds, the time necessary for the transmission of the foreign protein to the hypersensitive liver, the formation or liberation of histamine-like1 substances by this organ, and their subsequent transmission to the secondary reacting tissues of the body. In erythrocyte anaphylaxis the latent period is usually about fifteen seconds. It seems evident, therefore, that the characteristic precipitous fall in arterial blood pressure in erythrocyte anaphylaxis does not depend upon preliminary hepatic reactions, but is due to immediate reactions in certain extra-hepatic tissues. We have endeavored to test this conclusion by evisceration and dehepatization.
Monoclonal anti-idiotypic antibodies to an I-J interacting molecule inhibit suppression in an H-2 restricted way.
Authors: Jaureguiberry B , Liao L , Kuchroo V , Dorf M E , Diamond B
Published: December 2022

We have obtained 10 mAb from two independent fusions that are anti-idiotypic to the combining site of an anti-I-Jk antibody. These mAb block Ts cell function isn a genetically restricted manner in vitro and in vivo and recognize a determinant on macrophage membranes. In addition, they do not affect the I-Ek-restricted activation of a Th cell line specific for pigeon heart cytochrome c. We conclude that these mAb may recognize a molecule other than conventional I-Ek on cells interacting with Ts cells that is involved in mediating Ts activity. The molecule recognized may be a modified I-Ek molecule or a molecule not encoded by the genes encoding I-Ek.
Role of dendritic cells in the regulation of class I restricted cytotoxic T lymphocyte responses.
Authors: Boog C J , Boes J , Melief C J
Published: December 2022

Two class I MHC mutant mouse strains, bm14 and bm13, differ from the strain of origin B6 in one and three amino acids in the alpha 1 and alpha 2 domains of the H-2Db molecule, respectively. These alterations result in specific failure to generate a CTL (Tc) response to the male-specific Ag H-Y. Immunization and/or restimulation in vitro with syngeneic male dendritic cells (DC), expressing very high levels of class I MHC molecules, restored the H-Y-specific Tc response of bm14 but not of bm13 mice. Serologically Db determinants were lost in normal spleen cells of both mutants, because FACS analysis showed a decreased binding of Db domain-specific mAb. Although bm13 DC show a higher fluorescence than bm13 normal spleen cells it is still strongly reduced (30 to 50%) in comparison with B6 DC. Surprisingly, bm14 DC show an equally very strong binding compared with B6 DC with these mAb. The quantitative expression of class I molecules on APC thus appears to be a major determinant in the regulation of Tc responses. In addition, immunization with DC markedly influenced the target cell specificity of the ensuing Tc response. The combined data clearly demonstrate that besides the highly efficient class II-restricted presentation of Ag to Th, shown previously, DC are also superior in the presentation of Ag in the context of class I molecules to Tc. bm14 DC are capable of directly activating H-Y-specific Lyt-2+ Tc memory cells without the need for L3T4+ Th. These biologic effects of DC can at least in part be explained by their very high class I MHC expression. Moreover, these results reiterate that class I MHC Db mutants and different APC can be used to study the contribution of specific class I domains to Tc recognition and restriction specificity.
Interaction of CD2 with its ligand, LFA-3, in human T cell proliferation.
Authors: Bierer B E , Barbosa J , Herrmann S , Burakoff S J
Published: December 2022

Recently, it has been demonstrated that lymphocyte function-associated Ag (LFA-3) is a natural ligand for CD2 and that this receptor-ligand interaction functions in cell-cell adhesion. In this report, we demonstrate that LFA-3 plays a role in T cell activation. L cells were transfected with human genomic DNA and sorted for expression of LFA-3. We demonstrate that LFA-3+ L cells, together with anti-CD3 mAb or with suboptimal doses of PHA, stimulate proliferation of human peripheral blood T cells. Furthermore, thymocyte proliferation was induced by LFA-3+ L cells and suboptimal doses of PHA. Proliferation was inhibited by mAb directed against either CD2 or LFA-3. Stimulation of thymocytes by the combination of PHA and LFA-3+ L cells resulted in the increased expression of the IL-2R, as well as of the surface Ag 4F2, transferrin receptor, and HLA-DR. These data support the conclusion that LFA-3 plays a role in CD2-dependent T cell activation. LFA-3 is widely distributed and is expressed on all APC and target cells. Thus, the ability of the CD2/LFA-3 interaction to costimulate with an anti-CD3 mAb suggests that the CD2/LFA-3 interaction may be involved not only in an Ag-independent alternate pathway of T cell activation but also in Ag-specific T cell activation.