bluuebunny/crossref_metadata_2025 huggingface dataset:
Interactions between Human Polymorphonuclear Leukocytes and Influenza Virus
Authors: HENRICKS P. A. J. , VAN DER TOL M. E. , VERHOEF J.
Published: June 2006

The effects of influenza virus A (H3N2) on several functions of human polymorphonuclear leukocytes (PMN) were examined. Incubation of PMN with virus induced chemiluminescence, aggregation, and degranulation of the leukocytes. The amount of chemiluminescence generated increased from 1 × 106 to 6 × 106 cpm when 2.5 × 106 to 2 × 107 virus particles were added to 2.5 × 106 PMN. Maximal aggregation occurred within 2 min and the response depended on the amount of virus added to the PMN. Release of acid phosphatase by virus‐treated PMN was 62±12% within 1 h compared with 7 + 7% by control PMN (P<0.005). Incubation of PMN with influenza virus resulted in a diminished phagocytic activity of the phagocytes. PMN from a patient with chronic granulomatous disease were similarly affected. It was thus concluded that the observed defect in phagocytic activity was not due to the reactive oxygen species generated hy the PMN during incubation with virus.
Naturally Occurring, Spleen‐Associated Suppressor Activity of the Newborn Mouse
Authors: JADUS M.R. , PECK A.B.
Published: June 2006

Immature monocytes residing m the spleens of newborn nine less than 6–7 days of age are known to suppress T‐dependent; and T‐independent immune responses in vitro and in vivo. Suppression is mediated in part through products secreted In the newborn monocytes, In the present study. we have identified 3 monokines secreted by newborn, but not adult, monocytes correlating with materials possessing significant suppressor activity in vitro. These monokines have molecular weights of 5N. 58. 10.8. and 10 kilodaltons. appear to be proteins, and are insensitive to heat inactivation. The 58 kd material is antigenically distinct from alpha‐fetoprotein. while the lower molecular weight materials arc too large to be prostaglandins. Although these monokines can he shown to have activity in vitro, they are not as efficient as newborn monocytes in preventing alloreactivity in vivo.
An OKT4+ T‐Cell Population in Séezary Syndrome: Attempts to Elucidate Its Lack of Proliferative Capacity and Its Suppressive Effect
Authors: GOLSTEIN M. M. , FARNARIER‐SEIDEL C. , DAUBNEY P. , KAPLANSKI S.
Published: June 2006

We have previously reported a case ot Sezary Syndrome (SS), in which an OKT4‐ T‐cell population exhibited a defective response to non‐specific mitogens, and an ability In suppress lectin‐induced T‐cell proliferation and pokeweed mitogen (PWM)‐induced B‐cell differentiation of normal donor peripheral blood mononuclear cells (PBMC) We report now that resting Sézary cells (SC) were essentially negative for activation activation (Ag) detected by monoclonal antibodies (MoAb) BI. 49.9. anti‐Tac. OKT9. OKT10. and OKIal. After phytohaemagglutin (PHA) stimulation, all these Ag were expressed With the notable exception of OKT10. Further investigations of SC functions indicated that no inlerleukin 2 (IL.‐2) biological activity was detected in culture supernalanls ot SC costimulated with PHA and phorbol myristate acetate (PMA). Interestingly, such stimulated SC exhibited a marked capacity to absorb exogenous IL‐2 while remaining unable to proliferate. These data suggest that patient's unresponsiveness to PHA may be unrelated to IL‐2 as an extracellular growth signal, but may instead be due to a failure in a later cellular activation event, subsequent to the binding of IL‐2 to its receptors. Lack of T10 Ag expression may be involved as a cause or a consequence. Kinetic study of suppression of PHA‐induced T‐cell proliferation of normal PBMC revealed that inhibition occurred during the first 24 h; moreover we showed that it was not due to limitation of available IL‐2 since it persisted in excess of IL‐2: remarkably the growth of an IL.‐2‐dependent murine cell line was unaffected by the presence of SC Further, inhibition was also observed on IL.‐2 independent calcium ionophore A 23187‐indueed T ‐cell proliferation of normal PBMC Taken together, the data suggest that the target of suppressor activity is probably an important ohligarory intracellular event controlling DNA replication, which is common to both IL.‐2‐Independent and IL‐2‐dependent T‐cell activation processes Human T‐cell leukaemia/lymphoma virus I (HTLV‐1) related p.19 and p.24 Ag were absent on fresh and.10‐day cultured SC. suggesting the absence of HTI.V‐1 infection, although not ruling out a proviral integration m the SC DNA.
Studies on the Mechanism of Binding and Uptake of Immune Complexes by Various Cell Types of Rat Liver in Vivo
Authors: van der LAAN‐KLAMER S. M. , HARMS G. , ATMOSOERODJO J. E. , MEIJER D. K. F. , HARDONK M. J. , HOEDEMAEKER P. J.
Published: June 2006

Soluble heterologous immune complexes (IC) were used to study the mechanism of IC binding to rat liver in vivo. Binding of IC to the various cell types of the liver, endothelial cells, hepatocytes. and Kupffer cells, was only inhibited by aggregated swine immunoglobulins. Binding was not inhibited by the absence of complement components. Intravenous injection of asialoglycoproteins. to block the galactose receptor, could not prevent IC binding. We conclude that Fc receptors play an important role in the binding of soluble heterologous IC to hepatocytes. endothelial cells, and Kupffer cells.
Chicken T‐Cell Growth Factor: Use in the Generation of a Long‐Term Cultured T‐Cell Line and Biochemical Characterization
Authors: VAINIO O. , RATCLIFFE M. J. H. , LEANDERSON T.
Published: June 2006

Supernatants from concanavalin A (Con A)‐stimulated chicken spleen cells were used to generate a long‐term cultured cell line from antigen‐primed chicken peripheral blood leukocytes. This line has been kept in continuous proliferation in vitro for more than 25 weeks. Morphologically these cells were lymphoblastoid and expressed class I and class II antigens of the major histocompatibility complex as well asT‐cell (but not B‐cell or macrophage) antigens. In addition they contained no peroxidase or non‐specific esterase activity, neither were they phagocytic. Proliferation of the line was totally dependent on exogenous T‐cell growth factor (TCGF) activity provided by the Con‐A‐stimulated spleen cell supernatant, comparable with the proliferation of Con‐A‐induced T‐cell blasts. TCGF activity from the supernatant was absorbed both by the long‐term cultured T cells and by Con A blasts, demonstrating the presence of receptors for the same TCGF species on the two populations. We have used the long‐term cultured cell line to characterize chicken TCGF further The molecular weight of the biologically active fractions found by gel filtration on Sephadex G‐100 was approximately 13,000 and isoelectric focusing showed chicken TCGF to have a pI of pH 5.9. We propose that the TCGF described here is the chicken analogue to the mammalian interleukin 2.
Dissection of the Human Antigammaglobulin Idiotype System with Monoclonal Antibodies
Authors: POSNETT D. N. , WISNIEWOLSKI R. , PERNIS B. , KUNKEL H. G.
Published: June 2006

Murine monoclonal antibodies (MoAb) were prepared by immunizing mice with human monoclonal rheumatoid factors from patients with mixed cryoglobulinaemia. Indirect solid phase radioimmunoassay and haemagglutination inhibition were used to screen the MoAb. Reactivity patterns of MoAb were determined using (a) polyclonal and monoclonal IgM proteins, (b) monoclonal IgM proteins from patients with neuropathy, (c) monoclonal and polyclonal IgM antigammaglobulins, and (d) various isolated VkIIIb‐positive immunoglobulins. Several patterns were obtained: MoAb reacting with private idiotypic determinants, with VkIIIb‐related determinants, and with cross‐reactive idiotypes (CRI). Two MoAb of the second type reacted with VkIIIb‐positive immunoglobulins and light chains regardless of their antigen activity. Another MoAb of reacted with VkIIIb light chains only when in association withγ heavy chains. MoAb of the third type defined distinct CRI systems. One of these was restricted to antigammaglobulins and another also involved neuropathy‐associated monoclonal IgM proteins. All MoAb clearly reacted with a minor population of normal polyclonal IgM, except for the MoAb detecting private idiotypic determinants. Studies using inhibition of binding by enzyme‐linked immunosorbent assay showed that polyclonal IgM antigamaglobulins may carry a CRI determinant detected by one of the MoAb. This CRI system, defined by the MoAb Glo 86.3, was similar to but not identical with the previously described Wa CRI, which is widely prevalent among IgM antigammaglobulins of rheumatoid arthritis.
Characterization of Two Morphologically Distinct Leu‐7+ Cell Subsets with Respect to Leu‐15 Antigen
Authors: FERRARI G. C. MANARA. C. , SCANDROGLIO R. , ROCCHI G. , PAGANI L. , DE PANFILIS G.
Published: June 2006

in the present study the fine structures of Leu‐7‐Leu‐15+ and Leu‐7+‐Leu‐15 cell subpopulations were characterized by using an immunogold‐immunoperoxidase double labelling in electron microscopy. The densities of Leu‐15 antigenic sites on both E rosetting (Er) and non‐adherent/non‐E rosetting (non‐A/non‐Er) Leu‐15 positive cell surfaces were also evaluated by using an immunogold analysis in electron microscopy. A majority of Leu‐7+ cells co‐expressed I he Leu‐15 antigen and showed an ultrastructural pattern specific for mature natural killer (NK) cells, i.e. abundant cytoplasm with many organelles. numerous electron dense granules and irregular outline. On the other hand, a minority of Leu‐7‐ ceils did not express the Leu‐15 antigen and showed a clearly different ultrastructural feature in comparison to Leu‐7+‐Leu‐15+ cells T hus, the presence of the Leu‐15 antigen on Leu‐7 + cell surface corresponds to ultrastructural features specific to differentiated NK cells and may represent an expression of Leu‐7+cell differentiation. An alternative hypothesis may he that Leu‐7+‐Leu‐15‐ and l.eu‐7‐Leu 15 cells represent distinct cell lineages within non‐A/non‐Er Leu‐7+ cells. Finally, the results of the present study provide proof that Leu‐7+ antigen is more frequently represented on non‐A/non‐Er Leu‐L‐15+ cells than on Er Leu‐15+ cells.
Formation of the Functional Alternative Pathway of Complement by Human Monocytes in Vitro as Demonstrated by Phagocytosis of Agarose Beads
Authors: HETLAND G. , ESKHLAND T.
Published: June 2006

Native agarose beads (diameter 5–10 μ), activators of the alternative complement pathway, are slowly phagocytosed when incubated with human monocytes cultured under serum‐free conditions. Agarose beads preincubated with monocyte cultures and then transferred to new cultures are more easily phagocytosed than native heads. These results indicate that the phagocytosis of agarose beads depends on opsonization of the beads by one or several substances of monocyte origin. By using antihuman C3 antibodies. trypsin treatment, and sodium dodecyl sulphale washing, we were able to demonstrate C3b and iC'3b son the agarose beads. The molecules were covalently bound to the surface of the beads. We conclude that in vitro human monocytes produce and secrete the essential factors for activation and propagation of the alternative complement pathway (factors C3. B, D. H and I), which becomes evident with an external activator like agarose beads in the cultures. The activation of complement by agarose beads results in the attachment of C3b and iC'3b to the surface of the beads, which are‐then phagocytosed by means of C3b and iC3b receptors on Ihe monocytes.
Human Schistosomiasis: Deficiency of Large Granular Lymphocytes and Indomethacin‐Sensitive Suppression of Natural Killing
Authors: GASTL G. A. , FELDMEIER H. , KORTMANN C. , DAFFALLA A. A. , PETER H. H.
Published: June 2006

Twenty‐eight children infected with Schistosoma haematobium and S. mansoni were tested for natural killer (NK) cell activity in vitro using the myeloid/erythroid cell line K562 as target. In addition, the frequency of large granular lymphocytes (LGL) and the number of HNK‐1−lymphocytes were examined in peripheral blood. NK cell activity was found to be markedly reduced in most patients when compared with a group of healthy Caucasian individuals (P<0.005). Moreover, the impairment of NK activity clearly correlated with the intensity of infection, which was quantified by parasite ova excretion in stool and urine. Within the lymphocyte compartment the percentages of cells with the NK phenotype (HNK‐1+) were found to be normal, although the majority of patients exhibited decreased numbers of LGL (P<0.005). The absolute and relative frequencies of LGL and HNK‐1+ lymphocytes by no means correlated with the parasite load. In vitro results suggest an at least partly prostaglandin‐mediated and interferon‐resistant functional defect of NK cells.
Preferential Induction of Antigen‐Specific Contrasuppressor T Lymphocytes by Trinitrophenyl (TNP)‐Substituted Langerhans Cells
Authors: PTAK W. , PTAK M. , GRYGLEWSKI A.
Published: June 2006

We have tested the ability of trinitrophenyl (TNP)‐labelled antigen‐presenting cells (Langerhans cells (LC) and peritoneal macrophages (Mø)) administered intravenously to induce cells that mediate and regulate contact sensitivity. TNP‐Mφ fail to induce contrasuppressor cells (Tcs) but activate efferent T suppressor (Ts) cells. However, the activity of immune cells can be recovered after removal of Ly 2 Ts cells. In sharp contrast, TNP‐substituted purified LC produced a significant contact sensitivity reaction, which is roughly equivalent to that achieved by skin sensitization with picryl chloride. Immune cells from these animals were relatively resistant to suppression by antigen‐specific Ts cells, and we have found that their resistance is due to the presence of Ly 1, Vicia villosa lectin adherent, I‐J+ cells. The Tcs cell induced by TNP‐LC is indistinguishable by surface markers and function from Tcs cell found in mice injected with antigen‐antibody complexes. Both these Tcs cells are capable of protecting immune cells from the effects of suppression by antigen‐specific Ts cells if added in the proper sequence. Although they have identical surface phenotypes, it is not known at present whether or not Tcs cells induced by two different antigen presentations are identical. The possible reasons why LC are such potent inducers of contrasuppression are discussed.
The Host Component of the Popliteal Lymph Node Graft‐versus‐Host Reaction
Authors: ROLSTAD B. , FOSSUM S. , HUNT S. V. , FORD W. L.
Published: June 2006

We have examined the cellular changes taking place in rat popliteal lymph nodes undergoing a graft‐versus‐host (GvH) reaction. Lamination of immunoperoxidase‐stain lymph node sections, using a panel of mouse monoclonal antibodies directed against different rat lymphoid cell subsets, revealed a disorganization of the lymph node architecture with disappearance of the follicles, and an intermingling of T and B cells, so that no distinct T ‐ and B‐cell areas were visible any more. Since the GvH nodes showed a preferential accumulation of host B cells over host T cells (particularly over the W 3/25’T helper cell subset), we also investigated the requirements for host B cell activation. The popliteal lymph node GvH reaction was induced in (PVG x DA)F1 rats by the injection of PVG cells into one foot and by DA cells into the other foot, and then immunoglobulin kappa allotype marked PVG B cells from athymic donors were injected intravenously. The allotype marked B cells proliferated vigorously in response to the DAT cells, but much less in response to the PVG T cells. These results indicate that the massive B‐cell activation taking place in GvH reactions may require an alloantigen incompatibility between donor T cells and host B cells, and argue against non‐specific mitogenic induction of the B cells.
Local Administration of Various Cytostatic Drugs after Subcutaneous Immunization Enhances Delayed‐Type Hypersensitivity Reaction to Sheep Red Blood Cells in Mice
Authors: TAN B. T. G. , LIMPENS J. , KOKEN M. , VALSTER H. , SCHEPER R. J.
Published: June 2006

Delayed hypersensitivity reaction in mice was enhanced with various anti‐cancer drugs administered at the site of antigenic stimulation during ‐4 days following sensitization. The immunopotentiating effect of the presented local chemotherapy protocol is thought to result from impairment of a regulatory circuit, with activated suppressor T cells (Ts) as target rather than Ts precursors or Ts‐inducing antigen‐presenting cells.
Intestinal Mucosal Memory and Presence of Memory Cells in Lamina Propria and Peyer's Patches in Mice 2 Years after Oral Immunization with Cholera Toxin
Authors: LYCKE N. , HOLMGREN J.
Published: June 2006

The build‐up of long‐term immunological memory in the gut mucosal immune system may explain long‐lasting protection against new attacks of cholera in convalescents from natural disease. We have looked for gut mucosal antitoxin immunological memory and memory cells in mice after oral immunization with cholera toxin. Our results show that mice that were orally primed with cholera toxin and then boosted 2 years later with a single oral antigen dose mounted a rapid and vigorous IgA antitoxin response in the intestinal lamina propria. A specific secondary antitoxin response could also be elicited without any in‐vivo boosting by in‐vitro stimulation of isolated lymphocytes from the lamina propria. Peyer's patches, mesenteric lymph nodes, and spleen. The results provide evidence for the almost life‐long persistence of anti‐cholera toxin memory B cells (and perhaps also T cells) in the intestine and probably also recirculating cells, after oral immunization with cholera toxin. A functional antitoxic immune response may be boosted rapidly on renewed enteral exposure to cholera toxin by the stimulation of memory cells both in the lamina propria and in the Peyer's patches.
Natural Killer and T‐Cell Potentiation by Monoclonal IgG against Natural Killer Cell FcR(IgG) or the T3 Complex
Authors: JONDAL M. , KULLMAN C. , ALTER M.‐B. , LJUNGGREN K.
Published: June 2006

Treatment of human natural killer (NK) cells with monoclonal antibodies of the IgG isotype against NK cell‐FcR (IgG) increased lysis of most haematopoietic target cell lines with high or intermediate background NK susceptibility. Treatment of normal non‐adherent lymphocytes with an IgG anti‐T3 monoclonal antibody also increased lysis against the same target cells. Potentiating anti‐FcR antibodies rapidly modulated FcR activity and the capacity of the cells to act as antibody‐dependent killers, although such antibodies were demonstrable for a long time at the cell surface. Anti‐FcR treatment did not influence concanavalin A (Con A)‐dependent killing, in contrast to anti‐T3 treatment, which suppressed lectin‐dependent lysis but did not influence antibody‐dependent killing. The data is compatible with a ‘pro‐receptor’ theory for FcR in NK killing, staling that such receptors may function in the same way as the T3 complex interacts with specific T cell receptors.
Differentiation by Interleukin 2 of a Subpopulation of Human B Cells
Authors: GRILLOT‐COURVALIN C. , LABAUME S. , BROUET J.‐C.
Published: June 2006

We demonstrate here that purified recombinant interleukin 2 (IL‐2) induces a sizeable population of highly purified spleen B cells, by both negative and positive (cell‐sorting) selection to differentiate polyclonally into plasma cells. This effect is seen at concentrations of IL‐2 that are optimal for T‐cell growth (1–5 U/ml) and is inhibitable by anti‐Tac monoclonal antibody. In contrast, peripheral blood B cells show no or minimal terminal differentiation. When spleen B cells were separated into small and large cells, only the latter responded to IL‐2; since this state of activation is achieved in vivo, these data support the possibility that IL‐2 may be a physiological mediator in B‐cell differentiation.
Inhibition of Human Lymphocyte Transformation by Tomato Lectin
Authors: KILPATRICK D. C. , GRAHAM C. , URBANIAK S.J.
Published: June 2006

The lectin from tomato (Lycopersicon esculentum) fruits was found to be non‐mitogenic for human lymphocytes in culture and actually suppressed spontaneous DNA synthesis. It also inhibited the transformation of human peripheral blood Lymphocytes induced by recall antigens or allogeneic cells in vitro. This inhibition was most effective when the lectin was present from the beginning of the culture period, and could be abolished by the simultaneous addition of oligomers of N‐acetylglucosamine The tomato lectin was able to bind to several major lymphocyte cell surface glycoproteins, but not to the major histocompatibility (HLA) antigens. The binding of tomato lectin to lymphocytes could be inhibited by wheal germ agglutinin (WGA). but not by concanavalin A. Tomato lectin could agglutinate monocytes and B lymphocytes as well as T lymphocytes. Human serum used to supplement the culture medium supporting lymphocyte transformation was equally effective after passage through a tomato lectin‐Sepharose column. The inhibition of lymphocyte transformation brought about by tomato lectin was not stopped by exogenously added interleukin 1 and/or interleukin ‐2 even at very high concentrations.
Carrier‐Specific Induction of Suppressor Cells Controlling Anti‐Erythrocyte Autoantibody Production in Mice
Authors: WATT G.J. , RUSSELL J. , ELSON C. J.
Published: June 2006

Mice immunized with rat erythrocytes develop anti‐erythrocyte autoantibodies, distinct anti‐rat erythrocyte agglutinins, and suppressor‐inducer cells. which regulate the production of autoantibody but not anti‐rat erythrocyte agglutinins upon transfer to naive recipients. In this report, we have tried to determine the specificity of the suppressor‐inducer cells. CBA/N mice (which express an X‐linked genetic B‐lymphocyte defect) immunized with rat erythrocytes developed no autoantibodies but normal levels of anti‐rat erythrocyte antibodies and suppressor‐inducer cells, thereby suggesting that neither idiotypes on autoreactive B cells nor idiotypes on autoantibody itself, stimulate suppressor‐inducer cells. In contrast, rat erythrocyte‐primed spleen cells suppressed both a primary 2,4,6 trinitrophenyl (TNP) response and anti‐erythrocyte autoantibody production (but not anti‐rat erythrocyte antibodies) upon transfer to naive recipients and challenge with TNP‐rat erythrocytes. It is considered that the suppressor‐inducer cells are carrier‐specific and that they are not simulated by idiotypes on either autoantibody or autoreactive B cells
Interaction between Human IgD and Ricinus Agglutinin
Authors: MARCHES R. , GHETIE V.
Published: June 2006

The results show that monoclonal IgD reacts specifically with ricinus agglutinin (molecular weight 120.000) (RcA1). The reaction between IgD and RcA1 was inhibited by galactose and lactose but not by glucose, indicating that the interaction is mediated by galactose containing carbohydrate units located in or near the σ‐hinge region. Affinity chromatography on RcA1‐Sepharose 4B was successfully used lo isolate monoclonal IgD from the IgD myeloma plasma.
Activation of Human Monocyte and Natural Killer Cell‐Mediated Tumour Cell Killing by Two Dialysable Thymic Factors
Authors: HAMPRECHT K. , VÖTSCH W. , ANDERER F. A.
Published: June 2006

Components of calf thymus extract dialysable under acid conditions contained two natural killer (NK) cytotoxicity‐stimulating factors. CSFa and CSFb, which could be separated by ion exchange chromatography. NK cytotoxicity of human peripheral blood mononuclear cells (PBMC) against human K562 tumour cells was strongly enhanced after 72 h pre‐incubation with the factors. The CSFa/b‐specific stimulation of PBMC required the presence of monocytes. The cytotoxic effector cells activated during pre‐incubation of PBMC with the CSF were identified as monocytes and as NK cells present in the fraction of large granular lymphocytes (LGL). Selective cell depletion studies with LGL‐containing subpopulations (free of monocytes) allowed factor‐specific discrimination of the activated LGL. Pre‐incubation of PBMC with CSFa stimulated NK cytotoxicity of LGL (Leu 7+ 11; T8−), whereas pre‐incubation with CSFb resulted in stimulation of LGL (Leu 7+11+; T8−). The biological effects of CSFa and CSFb could be further distinguished by analysis of surface marker expression during incubation of PBMC. CSFb scarcely influenced T4 expression, hut strongly enhanced the expression of TK and that of transferrin receptor, whereas CSFa had no significant influence on the expression of these three surface markers. Both factors induced a drastic reduction of tumour take incidence or tumour development in mice when applied before and after tumour challenge.
Role of Infection in the Modulation of Mouse Circulating Soluble Cell‐Free Fcγ2b/γ1 Receptor
Authors: KHAYAT D. , SERBAN D. , DUX Z. , SCHLOMO Y. , JACQUILLAT C.
Published: June 2006

We have recently demonstrated the occurrence of functional circulating soluble cell‐free Fcγ2b/γ1 receptor in normal mouse serum by means of the 2.4G2 rat monoclonal antibody. With the solid phase radioimmunoassay described previously, we report here a dramatic increase in the level of circulating soluble cell‐free Fcγ2b/γ1 in Schistosoma mansoni‐ infected mice individually tested before and on days 22, 41, 62, and 82 after infection. This increase was statistically highly significant when the five measurements from each mouse were compared, and also when the entire group of infected mice was compared with the stable level of Cf‐ Fcγ2b/γ1R measured in strain‐, sex‐, and age‐matched control mice individually tested on the same days. This increase was commensurate with a rise in the amount of IgG detected in the sera of the S. mansoni‐infected mice. Thus, infection seems to be one of the factors that modulate the expression of such soluble Cf‐Fcγ2b/γ1R in mouse serum. Furthermore, the experimental device reported here for the production of high levels of Cf‐ Fcγ2b/γ1R by infection (in this case by parasitic infection) could serve as a model for obtaining large quantities of serum Cf‐ Fcγ2b/γ1R for further purification. Lastly, we point out that, by establishing a functional relationship with circulating IgG, such soluble Cf‐ Fcγ2b/γ1R may modulate some of the functions in which the Fc portion of immunoglobulins is involved.
Development of a Model System for Analysing Graft‐versus‐Host‐Mediated Immune Suppression
Authors: DEMOYEN P. LANGLADE , LARSSON E.‐L.
Published: June 2006

F1, hybrid mice injected with parental T lymphocytes undergo a graft‐versus‐host (GvH) reaction, one of the consequences being a highly depressed cytotoxic T‐lymphocyte (CTL) potential against alloantigens and modified self antigens. In the present study we demonstrate that concanavalin A (Con A) can he used for analysing such GvH‐mediated immune suppression. Thus, both alloantigen‐ and Con A‐induced responses were reduced by approximately 80% in F1, suppressed (F1s) animals as compared to F1 control (F1c) mice, Although interleukin 2 (IL‐2) production was found to be reduced by approximately 50% it did not account for the reduced CTL response in F1s mice. The addition of IL‐2 to Con A‐stimulated F1s spleen cell cultures did not reconstitute the response. The results suggest that the suppressive mechanism operates by preventing a large fraction of Lyt‐2+ CTL precursors from acquiring IL‐2 reactivity. However, a small fraction of CTL precursors, escaped the suppression and differentiated into effector CTL.
Natural Killer Cells Contribute to Inflammation but Do Not Appear to be Essential for the Induction of Clinical Lymphocytic Choriomeningitis
Authors: ALLAN J. E. , DOHERTY P. C.
Published: June 2006

The inflammatory exudate found in cerebrospinal fluid (CSF) of mice 6 days after intracerebral infection with lymphocytic choriomeningitis virus (LCMV) contains substantial populations of both cytotoxic T lymphocytes (CTL) and natural killer (NK) cells. Removal of NK cell activity by in vivo treatment with antibody to the asialo GM, ganglioside and studies with NK‐deficient bg/bg mice did not clearly determine whether NK cells contribute in any way to the development of clinical LCM. However, the LCM disease process induced in cyclophosphamide‐suppressed, LCMV‐infected recipients by the adoptive transfer of LCMV‐immune spleen cells occurs in the absence of NK cell effector function in spleen, lymph nodes, or CSF of the recipients, though potent CTL populations are present in all of these sites. In this situation, NK cells are apparently not required for the induction of neurological symptoms that are indistinguishable from those of classical LCM.
Inhibition of Drug‐Induced Eosinophilia by Cyclosporin A
Authors: THOMSON A. W. , MILTON J. I. , ALDRIDGE R. D. , DAVIDSON R. J. L. , SIMPSON J. G.
Published: June 2006

Administration of cyclophosphamide to Sprague‐Dawley rats 2 days before immunization with ovalbumin in complete Freund's adjuvant resulted in a striking blood, bone marrow. and tissue eosinophilia. The number of circulating eosinophils reached a maximum (50‐fold increase above normal) 2 weeks after immunization, and eosinophils were also prominent in bone marrow, lymph nodes (paracortical areas), spleen (while and red pulp), and liver. The eosinophilia could be inhibited by daily oral administration of cyclosporin A (CsA), although its effect was dependent both on the duration of treatment and on the dosage of CsA. A similar, inhibitory action of CsA was demonstrated with respect to methotrexate‐induced eosinophilia. This experimental model may prove useful in the study of factors regulating eosinophil production and in examining the prospective value of CsA in the treatment of conditions where eosinophils play a central role.
Release of Immune Complexes bound to Erythrocyte Complement Receptor (CR1), with Particular Reference to the Role of Factor I
Authors: JEPSEN H. H. , SVEHAG S.‐E. , JENSENIUS J. C. , SIM R. B.
Published: June 2006

The release of 125I‐bovine serum albumin (BSA)‐anti‐BSA immune complexes (IC) hound to human erythrocyte complement receptors (E‐CR1) was studied. IC were complement‐solubilized in normal human serum (NHS), and reacted with human erythrocytes at conditions optimal for binding of the IC to E‐CRI. E‐CRI‐bound IC could he released by the addition of NHS or purified factor I. Factor I‐deficient or I‐depleted serum mediated no release, and addition of purified factor 1 restored the release. Factor H was not required for the release of IC. The kinetics of IC release was influenced by the NHS concentration, the presence of EDTA, and the time of prior storage of the erythrocytes at 4°C. NHS (1:5 to 1:10) in the presence of EDTA caused nearly maximal release within 10–20 min at 37°C. In the absence of EDTA the NHS‐induced IC release was markedly slower. IC released within she first 30 min showed significant rebinding to new E. The release of IC was not associated with loss of the IC binding activity of E‐CRI. The NHS‐mediated release of IC could be inhibited by rabbit anti‐CRI and by a mixture of protease inhibitors. Release induced by purified factor 1 was also inhibited by protease inhibitors. The affinity of IC binding to E‐CRI was reduced alter cleavage of CRI‐bound C3b‐IC to iC3h‐IC by factor I.
A Sensitive Method for the Detection of Specific Antibody Production in Different Isotypes from Single Lamina Propria Plasma Cells
Authors: LYCKE N.
Published: June 2006

A sensitive and reproducible method for the detection of specific antibody production (or total immunoglobulin secretion) at the single cell level from isolated lamina propria lymphocytes was developed. The cells were prepared from mouse intestinal mucosa by enzyme extraction with collagenase, and antibody secretion was demonstrated with a solid phase enzyme‐linked immunospot (ELISPOT) assay. Oral immunizations with cholera toxin or keyhole limpet haemocyanin to mice gave high numbers of highly antigen‐specific spot‐forming cells (SFC) among isolated lamina propria lymphocytes. Spots were shown to result from active synthesis of immunoglobulin in vitro. The variation in SFC numbers between individual animals after a given protocol of oral immunizations was found to be 25% and between equal groups analysed on different occasions, 12%. Kinetics of primary as well as secondary immune responses after oral immunizations with cholera toxin were easily monitored. A single dose of cholera toxin gave rise to 230 antitoxin SFC/107 isolated lamina propria lymphocytes. Each additional dose stimulated to increasing numbers of specific SFC with roughly 7000 antitoxin SFC/107 cells after five immunizations. Monitoring of day‐by‐day responses after oral booster immunizations demonstrated peak SFC numbers on day 8 after antigen administration. The total number of immunoglobulin‐secreting (Ig) cells and the isotype distribution of specific SFC could also be determined. In the peak antitoxin response, 8% of the isolated total Ig‐secreting lamina propria cells were active against cholera toxin, and of these 80% were producing IgA. This method has also been successfully used in humans and rabbits to demonstrate specific antibody production by single lamina propria plasma cells.
Isolation and Characterization of Type IIa and Type IIb Fc Receptors from a Group A Streptococcus
Authors: YARNALL M. , BOYLE M. D. P.
Published: June 2006

Certain group A streptococcal strains have been reported to express two distinct type II receptors that bind to the Fc region of human IgG. In this study, we have isolated and characterized these two type II Fc receptors and characterized their reactivity with differing species of IgG. The type IIa receptor was found to he a 56,000 molecular weight protein which binds human IgG1, IgG2 and IgG4, in addition to pig and rabbit IgG. The type IIb receptor was found to he a 38, 000 molecular weight protein that bound exclusively to human IgG3. Neither the type IIa nor the type IIb receptor hound to goal, cow, dog, rat, or sheep IgG. Monospecific polyclonal antibodies were prepared against both the type IIa and type IIb Fc receptors These antibodies demonstrated that the type IIa and type IIb were anligcnically closely related and could not be distinguished from each other on the basis of their reactivity with either antibody. The distribution of type IIa and type IIb Fc receptors on a variety of different nephritogenie and non‐nephritogenic group A streptococcal strains was documented.
The Role of Calcium in Stimulation of Activated T Lymphocytes with Interleukin 2
Authors: LARSEN C. S. , KNUDSEN T. E. , JOHNSEN H. E.
Published: June 2006

In a study of the role of Ca in the stimulation of activated T lymphocytes with interleukin 2 (IL‐2) it was found that IL‐2‐induced proliferation can occur independently of extracellular calcium. Further, there was no correlation between triggering of DNA synthesis and an increase in free cytoplasmic calcium. However. IL‐2 induced an increased uptake of 45Ca from the extracellular medium. Since there is no increase in free cytoplasmic calcium, it must be assumed that this is caused by an increase in membrane‐associated calcium. Further, the calcium channel‐blocking agent, verapamil, and TMB‐8, a putative inhibitor of mobilization of calcium from intracellular pools, both exerted a dose‐dependent inhibition of IL‐2‐induced DNA synthesis in activated T lymphocytes. We conclude that calcium is not a second messenger in activated T lymphocytes stimulated by IL‐2, but our results indicate that calcium may play a role at membrane level.
Binding of Collagen Type II to Rheumatoid Synovial Cells
Authors: KLARESKOG L. , RUBIN K. , HOLMDAHL R.
Published: June 2006

Binding of biotin‐labelled native and denatured collagen type II and of aggregated IgG to frozen sections of synovial tissue from patients with rheumatoid arthritis (RA) or juvenile chrome arthritis (JCA) was investigated with the help of an avidin‐biotin‐peroxidase (ABC) technique. A large number of lymphocyte‐like and plasma cell‐like cells within the investigated biopsies aggregated IgG, and can be assumed to produce rheumatoid factors in five out of six cases a smaller number of lymphocyte‐like and plasma cell‐like cells bound native collagen type II. Denatured collagen type II bound mainly to cells within the synovial lining and to endothelial cells within the inflamed synovial tissues Binding of denatured but not of native collagen II was abolished by preincubation with rabbit antibodies towards human fibronectin. It is suggested that the method described here, using biotinylated antigens, may be of value for the study of local antibody production via investigations on frozen tissue sections, and that local antibody production against native collagen type II occurs within the inflamed synovial tissues at least in some cases of rheumatoid arthritis and juvenile chronic arthritis.
T‐Cell Helper Activity and B‐Cell Function of Synovial and Blood Lymphocytes from Patients with Rheumatoid Arthritis or Other Forms of Chronic Arthritis
Authors: NILSSON E. , VON STEDINGK L.‐V. , BIBERFELD G.
Published: June 2006

The helper effect of T cells on B‐cell immunoglobulin (Ig) responses induced by pokeweed mitogen (PWM) or purified protein derivative of tuberculin (PPD) was studied in lymphocytes from synovial fluid (SF) and blood of nine patients with rheumatoid arthritis (RA) and eight patients with other forms of chronic arthritis. In PWM cultures the helper effect of SF T cells on Ig responses (IgG, IgM, IgA) of autologous and allogeneic blood B cells was lower than that of blood T cells (P<0.01). This decrease was more pronounced in patients with RA than in patients with non‐RA. In PPD cultures no significant difference was found between the helper effect of SF T cells and blood T cells on the Ig responses of allogeneic blood B cells or on the IgG response of autologous blood B cells, whereas the helper effect of SF T cells on the IgM and IgA responses of autologous blood B cells was decreased. The Ig responses to PWM or PPD in cocultures of autologous blood B and T cells were not significantly different between patients and healthy controls. The PWM‐ and PPD‐induced Ig responses of SF B cells were lower than those of blood B cells when cocultured with autologous blood T cells. SF B cells produced IgG but usually little IgM and IgA. Thus there was a dysfunction of SF B cells and of SF T cells in a PWM‐driven system, but a fairly good helper function of SF T cells in a PPD‐driven system.
Cellular Location of ‘Liver‐Specific’ Antigen F
Authors: FORSTER M. , CELIO M. R. , WINTERHALTER K. H. , BINZ H.
Published: June 2006

Three mouse monoclonal antibodies against Type 1 murine “liver‐specific” antigen F and one rat monoclonal antibody against human antigen F were produced. These monoclonals recognize antigenic determinants common to Type 1 and type 2 mouse and human antigen F. By immunolabelling with β‐galactosidase. antigen F could be demonstrated in the cytoplasm of hepatocytes, tubules of the kidney, and perikarya of neurons of the central nervous system. F antigens isolated from liver, kidney, and brain all have molecular weights of around 40,000.
Demonstration of Species‐Specific Sensitization to Major Allergens of Dermatophagoides Species by Solid‐Phase Absorption of Human IgE Antibodies
Authors: LIND P. , INGEMANN L. , BROUVHZ M.
Published: June 2006

Twenty‐five mite‐allergic patients in the Brussels area were tested with the skin‐prick test (1.5 and 10 histamine equivalents by prick test (HEP) and RAST with standarized extracts of Dermatophagoides pteronyssinus (Dp) and D. mocroceras (Dm). Highly significant correlations (9.69<rS<0.96) between the two species coincided with marginal, but highly significant P0,01), greater reactions to Dp in all comparisons. Species‐specific IgE antibodies to antigen Dp‐42 (=P1) and Dp‐X were readily detected by solid‐phase serum absorption with heterologous antigen and subsequent crossed radioimmunoelectrophoresis (CRIE). An absorption single radial immunodiffusion (SRID) technique, involving major allergens Dp‐42, Dm‐6 and Df‐6 of Dp, Dm and D. farinae (Df). demonstrated the presence of IgE antibodies reacting with (1) epitopes common to all species. (2) epitopes common to two species, absent on the third, and (3) epitopes strictly specific for Dp. The majority of patients with a strong IgE binding reactecd to both common and strictly species‐specific epitopes. The serological findings were in good agreement with the exposure situation in the area: by species‐specific ELISA for dust allergen content. Dp occurred as the dominating species, 20 out of 24 homes having dust levels of Dp‐42 above 30 ng/g. Five and four homes had detectable levels of Dm‐6 and Df‐6. respectively, in dust.
The Roles of Influenza Virus Haemagglutinin and Nueleoprotein in Protection: Analysis Using Vaccinia Virus Recombinants
Authors: ANDREW M. E. , COUPAR B. E. H. , BOYLE D. B. , ADA G. L.
Published: June 2006

Vaccinia virus recombinants expressing haemagglutinin (HA) or nucleoprotein (NP) from influenza virus A/PR/8/4 were used to investigate protective immunity in mice, with two protocols. Protection was assessed by mortality and morbidity rates and by lung virus titres after infection intranasally with A/PR/8/34. In the first protocol, mice immunized with vaccinia‐HA recombinaant virus and infected intranasally with A/PR/8/34 were almost totally protected, but mice immunized with vaccinia‐NP virus were very poorly protected. In the second protocol, the recombinant viruses were used to stimulate in vitro T cells that are specific for HA and NP; both populations of T cells, when transferred to A/PR/8/34‐infected mice, afforded good protection. The results indicate that an immune response specific for just HA provided protection that was almsot indistinguishable from that provided by whole A/PR/8/34. On the other hand, immunization with vaccinia‐NP provided poor protective immunity, despite the fact that transferred NP‐specific T cells were very effective and vaccinia‐NP immunization has previously been shown to stimulate cytotoxic T cells. These results demonstrate that a single viral antigen, delivered by live vaccinia virus, can provide effective protection, but that immunization for cross‐protection against heterologous influenza virus remains elusive.
Methods for the Detection of a Specific Mycobacterium leprae Antigen in the Urine of Leprosy Patients
Authors: KALDANY RASHAD‐RUDOLF J. , MAASHO KERIMA , OHMAN R. , REITZ‐VICK D. , BRITTON S. , LEFFORD M. J.
Published: June 2006

Two methods for detecting the phenolic glycolipid, PGL‐1, a Mycobacterium leprae‐specific molecule, in the urine of leprosy patients are described. Both methods rely on the 100‐fold preconcentration of the urine, which can be accomplished by a single‐step ultrafiltration procedure. The equivalent of approximately 2.5/μ of PGL‐1/ml was detected in the urine of LL patients with an inhibition ELISA. The second method, a direct dot‐blot assay on nitrocellulose paper, was much simpler and more sensitive. As little as 3 ng of antigen was detected by the dot‐bolt technique. PGL‐1 was detected in the urine of LL patients.
Vaccination‐Induced Circulation of Human B Cells Secreting Type‐Specific Antibodies against Pneumococcal Polysaccharides
Authors: HEILMANN C. , HENRICHSEN J. , PEDERSEN F.K.
Published: June 2006

Indirect plaque‐forming cell assays detecting B cells secreting antibodies against capsular pneumococcal polysaccharide (PPS) antigens are described. In healthy adult volunteers the total number of B cells secreting IgM antibodies against the antigens in a polyvalent PPS vaccine reached a maximum in the blood 6 days after in vivo immunization (mean: 552/106 mononuclear cells), whereas the highest concentration of IgG and IgA antibody‐secreting cells (SC) were detected 7 days after immunization (means: 628 and 1691/106). B cells secreting antibodies to PPS type 3 (PPS3), PPS8, PPS18C and C‐polysaccharide (CPS)—a cell wall antigen common to all pneumococci—constituted 9%, 16%, 6% and 5% (means) of the total number of antibody SC respectively. While the majority of the anti‐PPS‐SC secreted IgA antobodies, the anti‐CPS‐SC almost exclusively secreted IgG. Pre‐vaccination concentrations of anti‐PPS were generally low in contrast to antibodies against CPS, which were present in high concentrations in all individuals. The discrepancy in the Ig class of the antibody SC is probably related to the difference in the pre‐vaccination immunity against PPS and CPS.
Spontaneous IgA Synthesis by Blood Mononuclear Cells in Alcoholic Liver Disease
Authors: VAN DE WIEL A. , SEIFERT W. F. , VAN DER LINDEN J. A. , GMELIG‐MEYLING F. H. J. , KATER L. , SGHUURMAN H. ‐J.
Published: June 2006

Changes in the concentration and composition of serum immunoglobulin A (IgA) and deposits of IgA in tissues are well‐known characteristics of alcoholic liver disease. We invesitigate whether these changes also accompany IgA synthesis by peripheral bloos mononuclear cells, (MNC), by counting immunoglobuliln‐producing cells using a solid‐phase enzymatic ‘spot’ test, and by analysis of immunoglobulin content in lysed cells with culture supernatant using conventional enzymatic methods. Patients with alcoholic liver disease exhibited a significantly higher number of spontaneously IgA‐producing cells than did normal healthy controls (1.7 × 106 cells/1 blood and 0.5 × 106 cells/1 blood, respectivly, P<0.01). The IgA content of MNC directly after isolation was also higher (38 and 13 ng/106 MNC, respectively, P<0.01), as was the IgA production during an unstimulated 6‐day culture period (520 and 95 ng/106 MNC put into culture, respectively, P<0.001).The spontaneously IgA‐producing cells assessed directly after isolation of mononuclear cells correlated with the IgA production during an unstimulated culture (P<0.01). We conclude that in alcoholic liver disease, B Iymphocytes circulate which may have been activated in vivio.
Neither Interleukin 2 nor Gamma Interferon Directly Promote Growth or Differentiation of Mouse B Cells
Authors: JULIUS M. H. , PAIGE C. J. , LEANDERSON T. , CAMBIER J. C.
Published: June 2006

The roles of interleukin 2 (IL‐2) and gamma interferon (IFN‐γ) as direct mediators of B‐cell growth and differentiation were analysed. Products of cloned genes were used in both cases. The use of flow cytometric assays coupled with density fractionation of reponding splenic B‐cell populations enabled both the characterization of B cells responding to various stimuli and the estimation of their frequency. B cells responding to no‐IL‐2 related lymphokines promoting growth and differentiation were restricted to low buoyant density fractions. In addition, these cells expressed densities of IL‐2 receptor determinants comparable to those found on T cells, although, IL‐2 did not support thier growth or differentiation. The inability to demonstrate any direct effect of either IL‐2 or IFN‐γ on B cells in any state of activation suggests that their physiological roles are mediated through additional cell types.
Enhancement and Suppression of an Intrastrain Cross‐Reactive Idiotype
Authors: MANZO C. , NISONOFF A.
Published: June 2006

The effects of a copolymer of monoclonal anti‐idiotype (7B7.10) with keyhole limpet heamocyanin (KLH), designated 7‐K, on an ongoing immune response were investigated. It was found that the response could be diverted to the production of higher of antip‐azobenzenearsnate (Ar) antibodies, of which nearly 100% carry an intrastrain crossreactive idiotype (CRIA). The effect was observed only in mice that had received a prreinoculation of KLH‐Ar, or KLH plus bovine gamma globulin‐Ar (BGG‐Ar). The effect was also observd, however, when cross‐linked 7B7.10 was mixed, rather than conjugated with KLH, suggesting that the role of KLH was to induce the production of a B‐cell growth factor. Cross‐linked 7B7.10 was not effective in the absence of KLH. A primary inculation of 7‐K together with KLH‐Ar did not result ini significant suppression or enhancement of CRIA. Also, pre‐inoculation of 7‐K alone did not suppress a subsequent idiotypic response to KLH‐Ar, whereas monomeric anti‐Id was suppressive. This supports a possible role for the unmodified Fc segment in the suppressive mechanism. In mice primed with KLH‐Ar, before administration of 7‐K, CRIA molecules lacking anti‐Ar activity were present in very low concentrations in the immune sera. The methods described permit the reproducible production of large amounts of CRIA anti‐Ar antibodies.