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1.	Gustafsson, Lars, et al. (author) Infectious disease, reproductive effort and the cost of reproduction in birds 1994 In: Philosophical transactions of the Royal Society of London: Series B. ; :346, s. 1655-1658 Journal article (pop. science, debate, etc.)abstract Reproductive effort can have profound effects on subsequent performance. Field experiments on the collared flycatcher (Ficedula albicollis) have demonstrated a number of trade-offs between life-history traits at different ages. The mechanism by which reproductive effort is mediated into future reproductive performance remains obscure. Anti-parasite adaptations such as cell-mediated immunity may probably also be costly. Hence the possibility exists of a trade-off between reproductive effort and the ability to resist parasitic infection. Serological tests on unmanipulated collared flycatchers show that pre-breeding nutritional status correlates positively with reproductive success and negatively with susceptibility to parasitism (viruses, bacteria and protozoan parasites). Both immune response and several indicators of infectious disease correlate negatively with reproductive success. Similar relations are found between secondary sexual characters and infection parameters. For brood-size-manipulated birds there was a significant interaction between experimentally increased reproductive effort and parasitic infection rate with regard to both current and future fecundity. It seems possible that the interaction between parasitic infection, nutrition and reproductive effort can be an important mechanism in the ultimate shaping of life-history variation in avian populations.
2.	Andersson, Måns Sverker, et al. (author) Glycosylated haemoglobin: a new measure of condition in birds 1995 In: Proceedings of the Royal Society of London. ; :260, s. 299-303 Journal article (peer-reviewed)abstract Abstract: The influence of condition on time of breeding and reproductive success has been discussed since Darwin first suggested a relation in 1871. We used a novel method to investigate the influence of condition on the timing of breeding and reproductive success by measuring a relatively inert physiological parameter - the amount of glycosylated haemoglobin - in blood samples taken from the collared flycatcher Ficedula albicollis. The percentage of glycosylated haemoglobin (%HbG) was assumed to be proportional to the average blood glucose level, during the 3-5 weeks before the blood sampling. The %HbG was influenced neither by sex nor age. Date of arrival at the breeding ground was negatively correlated with %HbG so that early-arriving birds had significantly higher %HbG than those arriving later. Clutch size, corrected for the effect of laying date, correlated positively with %HbG in females, as did the number of fledged young, corrected for the effect of laying date, for both sexes. We found no correlation between body mass and the %HbG. We suggest that prebreeding condition influences the timing of breeding and subsequent reproductive performance and that %HbG can be used as an indicator of prebreeding-condition in migrating birds.
3.	Schulze-Osthoff, Klaus, et al. (author) Redox Signaling by Transcription Factors Nf-Kappa-B and Ap-1 in Lymphocytes 1995 In: Biochemical Pharmacology. - 0006-2952 .- 1356-1839. ; 50:6, s. 735-741 Journal article (peer-reviewed)
4.	Dahlgren, Ulf, 1953, et al. (author) Expression of a dietary protein in E. coli renders it strongly antigenic to gut lymphoid tissue. 1991 In: Immunology. - 0019-2805. ; 73:4, s. 394-7 Journal article (peer-reviewed)abstract Bacteria that colonize the intestinal mucosa elicit a strong mucosal immune response, whereas food antigens such as ovalbumin are very weakly immunogenic to the gut-associated lymphoid tissue. This may either be due to special physico-chemical properties of bacterial substances versus proteins from animals and plants, or to stimulating properties of the bacteria on, e.g., antigen presentation, rendering all substances contained within bacteria antigenic. To test these hypotheses, ovalbumin was expressed in wild-type Escherichia coli and germ-free female rats were colonized with this strain. The systemic and mucosal antibody response of these rats was compared with that of rats given large amounts of dietary ovalbumin. Biliary IgA antibodies, which reflect the local IgA antibody production in the intestine, were only found in the rats colonized with ovalbumin-synthesizing E. coli. IgG antibodies in the bile were also only seen in these rats. We conclude that mucosal immunogenicity depends on the context in which a protein is presented to the gut-associated lymphoid tissue, rather than to special antigenic characteristics of the protein in itself.
5.	Dahlgren, Ulf, 1953, et al. (author) The secretory antibody response in milk and bile against fimbriae and LPS in rats monocolonized or immunized in the Peyer's patches with Escherichia coli. 1990 In: Immunology. - 0019-2805. ; 71:2, s. 295-300 Journal article (peer-reviewed)abstract The homing of lymphoid cells to mucosa-associated lymphoid tissue is, amongst other factors, influenced by the nature of the antigen used to induce an immune response. To study this phenomenon we have monocolonized rats with a type 1 fimbriated Escherichia coli O6K13H1 strain and compared the secretory antibody response to colonization with the primary and secondary response obtained in rats immunized in the Peyer's patches (PP). Samples were tested with respect to the titres of antibodies against two antigens present on the E. coli strain: O6 lipopolysaccharide (LPS) and type 1 fimbrial antigen. In the primary immunized animals, IgA anti-fimbrial antibodies were mainly seen in milk and IgA anti-LPS antibodies mostly found in bile. In the booster immunized, and in the monocolonized, animals there was a shift of the antibody response towards the bile. Thus anti-fimbrial antibodies appeared in milk at approximately the same or at a lower level than in bile and the IgA anti-LPS antibodies were almost completely absent in the milk. The IgG antibody response of the animals immunized in the PP was primarily confined to milk for both anti-LPS and anti-fimbrial antibodies, while the colonized animals responded with higher levels in bile than in milk. IgM antibodies were only seen in the milk, except in primary immunized animals in which biliary IgM antibodies also were found. The data illustrate that: (i) primary stimulated cells predestined to produce IgA anti-LPS antibodies home mainly to the intestine, while cells predestined to anti-fimbrial antibody production have a greater tendency to populate the mammary gland; (ii) after repeated antigen stimulation and maturation of the immune response the cells are directed from the mammary gland to the intestine. We thus conclude that the nature of the antigen and the stage of lymphocyte maturation influences the homing of the cells and the appearance of various antibodies in different secretions.
6.	Dahlman-Höglund, Anna, 1964, et al. (author) Induction of IgE antibodies and T-cell reactivity to ovalbumin in rats colonized with Escherichia coli genetically manipulated to produce ovalbumin. 1992 In: Immunology. - 0019-2805. ; 76:2, s. 225-8 Journal article (peer-reviewed)abstract The immune response to ovalbumin (OA) and the bacterial antigens, lipopolysaccharide (LPS) and fimbriae were studied in conventional rats colonized from birth with an Escherichia coli strain producing OA. The colonized rats had developed IgE antibodies against OA, but not against the fimbrial or the LPS antigens from the E. coli at 2 months of age. At this time all rats were primed with OA given intracutaneously in Freund's complete adjuvant. Two weeks later the colonized rats showed a 35% greater delayed-type hypersensitivity (DTH) reaction to OA, measured as ear swelling, than the controls. Thus bacteria carrying antigens resembling potential allergens might aggravate, or participate in the induction of allergic symptoms. In addition such bacteria could be efficient vaccine vectors in protection against parasites. The study illustrates the importance of the mode of antigen presentation for the subsequent immune response.
7.	Gustafson, Rolf, et al. (author) Prevalence of Borrelia burgdorferi sensu lato infection in Ixodes ricinus in Sweden 1995 In: Scandinavian Journal of Infectious Diseases. - : Informa UK Limited. - 0036-5548 .- 1651-1980. ; 27:6, s. 597-601 Journal article (peer-reviewed)abstract Between 1988-1991, a total of 3,141 Ixodes ricinus ticks, 2,740 adults and 401 nymphs, was collected from different localities in 23 of the 25 provinces of Sweden. The ticks were identified, dissected and examined for the presence of Borrelia spirochetes. Indirect immunofluorescence was performed, using an antiserum obtained from rabbits, immunized with sonicated, whole Borrelia burgdorferi spirochetes isolated from Swedish Ixodes ricinus ticks. Borrelia-positive I. ricinus were found in all 23 provinces. The prevalence of infection in adults ranged from 3% in Jämtland to 23% in Södermanland. In nymphs, the infection prevalence ranged from 0% in 9 provinces to 15% in Södermanland. A significantly greater proportion of the adult ticks were found to be positive for Borrelia in the southern and central parts of Sweden as compared to the northern part (Norrland). No significant difference in prevalence could be demonstrated between the western and eastern parts of Sweden. On average, 10% of the nymphs and 15% of the adult I. ricinus were positive for Borrelia. Of 41 non-I. ricinus ticks, none was positive for Borrelia. This study shows that Borrelia burgdorferi sensu lato is present throughout the distributional area of I. ricinus in Sweden. This should lead to increased awareness of signs and symptoms compatible with Lyme borreliosis in persons living in or visiting areas where I. ricinus is present.
8.	Herías, M V, et al. (author) Increased antibody production against gut-colonizing Escherichia coli in the presence of the anaerobic bacterium Peptostreptococcus. 1998 In: Scandinavian journal of immunology. - : Wiley. - 0300-9475 .- 1365-3083. ; 48:3, s. 277-82 Journal article (peer-reviewed)abstract Germ-free rats were colonized with E. coli alone, or with E. coli plus Lactobacillus acidophilus and a strain of the obligate anaerobic gram-positive species, Peptostreptococcus. The presence of Peptostreptococcus reduced translocation of E. coli, but increased the serum antibody response to E. coli antigen. Whereas the immunoglobulin G (IgG) anti-E. coli antibodies largely represented cross-reactive antibodies, those of the immunoglobulin M (IgM) isotype represented true anti-E. coli antibodies because they could not be absorbed by L. acidophilus or Peptostreptococcus but could with E. coli. We suggest that peptostreptococci prime the gut immune system to other bacterial antigens and that this could be a mechanism behind the reduced translocation of facultative anaerobes in the presence of obligate anaerobes.
9.	Los, Marek Jan, et al. (author) Hydrogen-Peroxide as a Potent Activator of T-Lymphocyte Functions 1995 In: European Journal of Immunology. - : Wiley. - 0014-2980 .- 1521-4141. ; 25:1, s. 159-165 Journal article (peer-reviewed)abstract During inflammatory processes infiltrating cells produce large amounts of reactive oxygen intermediates (ROI). Increasing evidence suggests that ROI besides being cytotoxic may act as important mediators influencing various cellular and immunological processes. In this study, we have investigated the effects of hydrogen peroxide on several aspects of lymphocyte activation. In ESb-L T lymphoma cells, micromolar concentrations of hydrogen peroxide rapidly induced activation of the transcription factor NF-kappa B, whereas DNA-binding activity of the transcription factor AP-1 was virtually not affected. In addition, hydrogen peroxide induced early gene expression of interleukin-2 (IL-2) and the IL-2 receptor alpha chain. The stimulation of IL-2 expression was found to be conferred by a kappa B-like cis-regulatory region within the IL-2 gene promoter. In contrast to these activating effects, addition of hydrogen peroxide was largely inhibitory on cell proliferation which is consistent with a general requirement of thiol compounds for lymphocyte proliferation. However, hydrogen peroxide significantly increased T cell proliferation when applied for a short period under reducing conditions. These data indicate that ROI may act as an important competence signal in T lymphocytes inducing early gene expression as well as cell proliferation.
10.	Mejlon, Hans, et al. (author) Seasonal prevalence of Borrelia burgdorferi in Ixodes ricinus in different vegetation types in Sweden 1993 In: Scandinavian Journal of Infectious Diseases. - : Informa UK Limited. - 0036-5548 .- 1651-1980. ; 25:4, s. 449-456 Journal article (peer-reviewed)abstract The aim of this investigation was to estimate the seasonal risk of contracting human Lyme disease in different vegetation types in southern Sweden. Host-seeking Ixodes ricinus ticks (Acari: Ixodidae) were collected with standardized methods during May-September 1988 and March-October 1989 at 10 different sampling sites. Tick abundance was greatest during May-June and August-September. Prevalence of Borrelia burgdorferi infection of the ticks was assessed by phase-contrast microscopy complemented by immunology. Spirochetal infection of tick larvae was not detected. The prevalence of infection among nymphal ticks differed significantly between years and between sampling sites. Infection prevalence was greater in adult females than in nymphs, but was similar in female and male ticks. Among all vegetation types studied, the greatest Lyme disease risk was deemed to be from I. ricinus nymphs during May and September in mixed forest vegetation.
11.	Mincheva-Nilsson, Lucia, et al. (author) Immunomorphologic studies of human decidua-associated lymphoid cells in normal early pregnancy. 1994 In: Journal of Immunology. - 0022-1767 .- 1550-6606. ; 152:4, s. 2020-32 Journal article (peer-reviewed)abstract Human decidual lymphocytes from early, normal pregnancy were characterized in situ with respect to ultrastructure and distribution of subsets. The ultrastructure of isolated decidual gamma delta T cells was also studied. CD45+ cells comprised 11 +/- 2% of all decidual cells. The majority were localized in large lymphoid cell clusters (LCC), near endometrial glands, or as intraepithelial lymphocytes (IEL) in glandular epithelium. The major cell populations in LCC were CD56+TCR-gamma delta+ cells, CD56+ cells, TCR-alpha beta+CD4+ cells, and TCR-alpha beta+CD8+ cells. All expressed activation markers (CD45RO, Kp43, and/or HML-1) and MHC class II Ag (HLA-DR, HLA-DP, and/or HLA-DQ). No B cells were found. Almost all IEL were activated TCR-gamma delta+ cells (CD56+ and CD56-). The glandular epithelial cells expressed heat shock protein 60 at the basolateral side facing the TCR-gamma delta+ IEL. Decidual lymphocytes displayed cytoplasmic processes, microvilli, characteristic cytoplasmic granules, and had intimate contact with neighboring cells. Lymphocytes in the outer rim of LCC and the stroma showed signs of cellular movement. Two main morphotypes of gamma delta T cells could be distinguished. One had single microvilli, membrane-bound granules, and nuclear inclusions. The other had many microvilli, nonmembrane-bound granules and cytoplasmic multivesicular bodies. Our data suggest that LCC are centers of immune reactivity where T and NK cells become activated. The activated cells may guard against infections and undue trophoblast invasion and/or be involved in modulating the local maternal immune system toward unresponsiveness against the semiallogeneic fetus.
12.	Grage-Griebenow, E., et al. (author) An Fcγ receptor I (CD64)-negative subpopulation of human peripheral blood monocytes is resistant to killing by antigen-activated CD4-positive cytotoxic T cells 1997 In: European Journal of Immunology. - : Wiley. - 0014-2980 .- 1521-4141. ; 27:9, s. 2358-2365 Journal article (peer-reviewed)abstract It has been demonstrated that in monocyte/T cell co-cultures activated with recall antigens, cytotoxic T cells were generated which are able to reduce the number of antigen-presenting monocytes. In previous studies we could show that a minor subset of monocytes, the Fc gamma receptor I-negative (CD64(-)) monocytes, exhibits significantly higher antigen-presenting capacity than the main population of monocytes (> 90%) which are Fc gamma receptor I-positive (CD64(+)). Therefore, we addressed the question whether they are also differentially susceptible to T cell-mediated killing. In the present study we demonstrate that the CD64(-) monocyte subset is more resistant to killing by antigen-activated T cells than CD64(+) monocytes, as indicated by a higher viability and recovery of CD64(-) monocytes. This mechanism involves CD95 (Fas) antigen, since monocyte death in co-cultures with antigen-activated T cells could be partially reduced by blocking anti-Fas monoclonal antibodies (mAb). In agreement with this finding, although CD95 antigen was expressed on CD64(+) and CD64(-) monocytes at comparable levels, killing of CD64(-) monocytes by activating anti-Fas mAb was lower than of CD64(+) monocytes.
13.	Lundqvist, Carina, et al. (author) Isolation of functionally active intraepithelial lymphocytes and enterocytes from human small and large intestine 1992 In: JIM - Journal of Immunological Methods. - 0022-1759 .- 1872-7905. ; 152:2, s. 253-263 Journal article (peer-reviewed)abstract A mild purification method has been developed for the isolation of human intraepithelial lymphocytes (IEL) and enterocytes from the same individual. The isolation procedure includes mechanical disruption of the mucosal layer, treatment with reducing agent and sedimentation followed by Percoll gradient centrifugation. Finally, epithelial cells are removed from the IEL fraction using magnetic beads coated with the anti-epithelial antigen monoclonal antibody (mAb) BerEP4. Leucocytes are removed from the enterocyte fraction using magnetic beads coated with mAbs directed against common leucocyte antigen (CD45). Using this procedure IEL and enterocytes have been isolated from apparently normal jejunal, ileal and colonic tissue specimens. Recoveries of IEL were 7 x 10(5), 4 x 10(5) and 1 x 10(5)/cm2 mucosa from jejunum, ileum and colon respectively. 1-2 x 10(6) enterocytes/cm2 mucosa were recovered from small intestine while the corresponding value for colonic biopsies was approximately 2 x 10(5) enterocytes/cm2. The IEL fraction was pure as judged by the low percentages of B cells, macrophages and BerEP4 positive cells (less than 4%) present in the purified fraction. The enterocyte fraction contained less than 2% CD45+ cells. The two cell fractions were viable and expanded in vitro. Enterocytes expanded spontaneously while IEL required initial stimulation with mitogens. The isolation procedure described here will make it possible to study the function of human IEL, interactions between IEL and enterocytes and the role of both cell types in local immunity.
14.	Wold, Agnes E, 1955, et al. (author) Characterization of IgA1, IgA2 and secretory IgA carbohydrate chains using plant lectins. 1995 In: Advances in experimental medicine and biology. - 0065-2598. ; 371A, s. 585-9 Journal article (other academic/artistic)
15.	Wold, Agnes E, 1955, et al. (author) Lectin receptors on IgA isotypes. 1994 In: Scandinavian journal of immunology. - 0300-9475. ; 39:2, s. 195-201 Journal article (peer-reviewed)abstract It has been shown previously that secretory IgA interacts with the mannose-specific lectin of Escherichia coli. The purpose of the study described here was to evaluate whether the N-linked oligosaccharide chains of the human IgA isotypes IgA1 and IgA2 differ in lectin receptor activity. A range of plant lectins specific for N-linked oligosaccharide chains were tested for their ability to precipitate IgA1 and IgA2 myeloma proteins, secretory IgA and free secretory component. IgA2 myeloma proteins reacted more strongly than IgA1 with the mannose-specific lectin ConA, whereas IgA1 myeloma proteins reacted more strongly than IgA2 with two galactose-specific lectins, Ricinus communis agglutinin I and Abrus precatorius agglutinin. This suggests that IgA2 possesses a larger proportion of short truncated complex type oligosaccharide chains and/or oligomannose type chains than IgA1. Further, IgA2 reacted more strongly than IgA1 myeloma proteins with Lens culinaris (lentil) lectin, and Pisum sativum (pea) lectin, suggesting that IgA2 exposes more of short, complex type chains fucosylated on the core than IgA1. The differences demonstrated in receptor activity between IgA1 and IgA2 may be important in their interaction with the microbial flora, as well with endogenous lectins, such as phagocyte receptors.
16.	Abu Bakar, Siddique, 1961- (author) Immunological and functional properties of a non-repeat region in Plasmodium falciparum antigen Pf155/RESA 1998 Doctoral thesis (other academic/artistic)abstract Plasmodium falciparum malaria is one of the most prevalent of all the great insect-borne diseases causing high morbidity and mortality in humans. As conventional anti-malarial treatment is becoming increasingly inefficient, alternative approaches to combat the parasite, like vaccine development, are of high priority. Among other antigens, Pf155/RESA has been taken into consideration for inclusion in a vaccine against the asexual blood stages of P. falciparum. Repeat sequences of this antigen induce neutralizing antibodies and are potentially involved in protective immune responses. However, the genetic restriction in the immune responses to these epitopes has made it of importance to define additional regions in this antigen which may be suitable for inclusion in a vaccine. The present studies were aimed at investigating potential immunodominant B- and T-cell epitopes in the non-repeat regions of Pf155/RESA suitable for inclusion in subunit vaccine immunogens.In order to define immunodominant B- and T-cell epitopes, short overlapping peptides were synthesized, corresponding to three different non-repeat sequences of Pf155/RESA. Sera collected from malaria endemic countries recognized pepetides representing N-terminal sequences of the antigen. Based on high antibody reactivity in ELISA, two peptides were selected for affinity purification of antibodies from Liberian sera. The purified antibodies were shown to be efficient in parasite growth inhibition in vitro. Furthermore, the peptides had the ability to stimulate peripheral blood mononuclear cells from P. falciparum-immune donors to proliferate and to secrete IL-4 and/or IFN-g. Although the frequency of responders was high, the magnitude of the responses was generally low.Antibodies were raised in rabbits against synthetic peptides corresponding to different non-repeat sequences in Pf155/RESA. Although all antisera reacted strongly with the corresponding peptide, they reacted only weakly with full-length Pf155/RESA. Antibodies to some of the non-repeat sequences inhibited merozoite invasion in vitro, notably, also the invasion of parasites deficient in Pf155/RESA, indicating the presence of an antigen highly homologous to Pf155/RESA.Upon infection with P. falciparum, erythrocytes expose cryptic determinants in band 3, which are thought to mediate cytoadherence to endothelial cells. The N-terminal non-repeat region of Pf155/RESA contains a hexapeptide motif which is highly homologous to the cytoadherence related sequences of erythrocyte band 3. Synthetic peptides containing this hexapeptide motif inhibited the binding of P. falciparum-infected erythrocytes to melanoma cells in vitro. Antibodies raised against the largely overlapping sequences displayed highly different specificity patterns. Antibodies to the cytoadherence related motif of Pf155/RESA as well as antibodies raised against the peptides corresponding to the band 3 motif inhibited cytoadherence but not parasite growth. In contrast antibodies to sequences adjacent to the Pf155/RESA cytoadherence motif inhibited parasite growth in vitro but had no effect on cytoadherence.In summary, although the target antigens for the anti-parasitic activities displayed by the antibodies in these studies are unclear, sequences in non-repeat regions of Pf155/RESA may induce antibodies with capacity to inhibit parasite growth in vitro or to block the cytoadherence of P. falciparum infected erythrocytes to endothelial cells. The combination of non-repeat sequences with repeat sequences of Pf155/RESA in a subunit vaccine may provide immunogens with an improved capacity to induce parasite neutralizing antibody responses.
17.	Ahlborg, Niklas, 1962- (author) Immune responses to repeat sequences of the Plasmodium falciparum malaria antigen Pf332 1995 Doctoral thesis (other academic/artistic)
18.	Andersson, Gudrun, 1951- (author) Monoclonal antibodies to human interferon-α and interferon-γ : Characterization and application with focus on interferon-γ as a T cell cytokine 1992 Doctoral thesis (other academic/artistic)abstract The major functions of the immune system are to combat infections and to control that normal auto-immunity does not turn into auto-aggression. Antibodies, together with T cell receptors and MHC molecules, dictates the immunological specificity. However, soluble factors (like cytokines and enzymes) and interactions between leucocytes and endothelium (via adhesion molecules) are decisive for the qualitative and quantitative nature of the immune response to a provocation.The aims of this study were to establish and characterize mouse monoclonal antibodies (mAbs) to the human cytokines interferon alpha (IFNa) and interferon gamma (IFNy), to develop and evaluate the applicability of immunoassays based on these mAbs, and to study T cell secretion of IFNy in response to antigen and in extracellular matrix interactions in vitro.Reactivity of the established mAbs with their respective natural antigen was ascertained and mAbs binding to distinct epitopes identified. We found individual binding patterns of the mAbs to IFNa variants. These varied if the IFNa subtypes were in solution or bound to different solid supports, underlining the importance of thorough characterization of mAbs. The applicability of the anti-IFNa mAbs in ELISAs and their use for characterization of natural IFNa mixtures was demonstrated.Of the mAbs established to IFNy two had suitable characteristics for a sensitive sandwich ELISA, which was also adapted for determinations of IFNy in body fluids as evaluated with serum and plasma. This ELISA combination was also applied in an ELISPOT assay for determination of IFNy secretion on the single cell level. Using the ELISA we could show that IFNy is a quantitative but also qualitative marker for cellular immunity in vitro as evaluated with Francisella tnlarensis as a model antigen (IFNy is required for clearance of this infection). Furthermore, appropriate culture conditions for future studies of IFNy as a marker for immunological memory in short term cultures were defined.Using purified human T cells suboptimally activated through the TcR/CD3 complex with immobilized OKT3 we found that co-immobilized fibronectin (Fn) could potentiate their secretion of IFNy as measured in ELISA and on the single cell level with the ELISPOT. This potentiation was inhibited with antibodies to the integrin VLA-5. IFNy has been shown to be an important factor in lymphocyte migration and Fn is an extracellular matrix component and abundant in most tissues.The finding that Fn can augment IFNy secretion may thus provide new insights into the migration dependent effector functions of T cells.
19.	Bayard, Cecilia, 1966- (author) Purification, characterization and immunological studies of rat urinary proteins causing allergy in humans 1999 Doctoral thesis (other academic/artistic)abstract Rats are among the most frequently used laboratory animals and allergy to them constitutes a common occupational problem. Approximately 20-30% of the persons engaged in work with laboratory animals acquire symptoms of allergy. These include rhinitis, conjunctivitis, urticaria and sometimes asthma, symptoms which usually develop during the first three years of exposure. The allergic reaction arises from direct contact with rat urine, or from airborne dusts originating from dried animal urine. Rat urine contains a complex mixture of proteins some of which are known to be allergenic, for example a2u-globulin. In urine from fertile male rats a2u-globulin is one of the most abundant proteins.The primary aim of this thesis was to purify and identify the most potent allergens in rat urine. Optimization of various methods for evaluation of the allergenicity of the identified allergens was the second aim. The third aim was to examine in some detail the IgE binding regions of the major allergen, Rat n 1.02, a2u-globulin.Rat urinary proteins were separated and purified by ultrafiltration and high resolution chromatographic methods. The two dominating proteins were identified in this study as different forms of the same parent protein, i.e. a2u-globulin. It was also proved that the urinary protein previously named pre-albumin had no amino acid sequence resemblance to prealbumin (transthyretin) present in rat serum.Rat urinary proteins separated by SDS-PAGE were used as antigens when IgE antibodies in sera from allergic patients were studied with immunoblotting. A detection system utilizing chemiluminescence and a luminometer apparatus gave scanning curves of the IgE-binding reactivities and allowed arbitrary measurement of the amount of IgE bound to certain urine proteins. Specific immunoblotting patterns of IgE binding to the electrophoretically separated proteins were shown for each individual. Differences were seen in regard of the distribution among the recognized antigens and the amount of IgE bound. All rat allergic subjects studied had IgE antibodies binding to the major urinary protein, a2u-globulin. However, this protein was not always the most dominating allergen.Overlapping octapeptides corresponding to the amino acid sequence of the major allergen Rat n 1.02 were synthesized on solid support and screened in parallel to pinpoint the IgE binding regions of the protein using a modified ELISA procedure. Our results indicate the existence of linear IgE-binding epitopes, mainly located towards the N-terminal and C-terminal parts of the protein, as recognized by IgE antibodies in the studied sera. The role of these short amino acid sequences in the allergic reaction and their appropriateness for immunotherapy calls for further investigation.In conclusion, this thesis has contributed to the isolation, identification and characterization of the allergens involved in a major occupational disease among people who work with laboratory animals, namely rat allergy.
20.	Brenden, N., et al. (author) Differential MHC expression requirements for positive selection of separate TCR Vb families 1999 In: Immunogenetics. - : Springer Science and Business Media LLC. - 0093-7711 .- 1432-1211. ; 49:1, s. 1-6 Journal article (peer-reviewed)abstract Positive selection has been proposed to be involved in protection from diabetes. We examined positive selection by fluorescence-activated cell sorter analyses in thymocytes of protected and susceptible E-transgenic and non-transgenic NOD mice. Three Vb families showed positive selection in E-transgenic mice. Vb6+CD4+ and Vb10+CD4+ thymocytes were found at higher frequencies in both protected NOD-Ea and susceptible NOD-DY mice. The increased frequencies of Vb13+CD8+ thymocytes were found in protected NOD-Ea mice only, and not in susceptible NOD-DY transgenic mice. These three Vb families were further examined in bone-marrow chimeras between NOD-Ea and non-transgenic NOD mice, where we could examine the contribution of E-expressing bone-marrow-derived cells in positive selection. We find that NOD-Ea→NOD-Ea chimeras have an increased positive selection of Vb13+CD8+ cells and that positive selection is more efficient when both thymic epithelium and bone-marrow-derived cells express the E molecule. This was also seen for Vb6+CD4+ cells. However, for Vb6, bone-marrow-derived cells alone were also capable of positive selection. Positive selection of Vb10+CD4+ cells was restricted to E-expressing thymic epithelium only. For Vb13+CD8+ cells, we found that positive selection is most efficient with E-expression on both thymic epithelium and bone-marrow-derived cells, although positive selection also occurs with E-positive epithelium only. For Vb6+CD4+ cells, the dominating selecting cells are bone-marrow-derived cells, and Vb10+CD4+ cells seem to be selected exclusively by the thymic epithelium. Thus, the conditions for positive selection seem to vary considerably between different Vb families.
21.	Brenden, N., et al. (author) Disease-protected major histocompatibility complex Ea-transgenic non- obese diabetic (NOD) mice show interleukin-4 production not seen in susceptible Ea-transgenic and non-transgenic NOD mice 1998 In: Immunology. - : Wiley. - 0019-2805 .- 1365-2567. ; 95:1, s. 1-7 Journal article (peer-reviewed)abstract The non-obese diabetic (NOD) mouse is an animal model for insulin- dependent diabetes that has many similarities to the human disease. NOD mice transgenic for the Ea gene, allowing expression of the E molecule, are protected from diabetes and rarely develop insulitis. An Ea transgene mutated in the promoter region, (ΔY) lacks E expression on most B cells, thymic medullary epithelium and primary antigen-presenting cells, and confers no protection whatsoever. We have used these transgenic NOD mice, together with non-transgenic NOD mice, to study the correlation of E expression and production of interleukin-4 (IL-4) and interferon-γ (IFN-γ). We show that protected E-transgenic NOD mice have elevated levels of IL-4 compared with non-transgenic mice, both in the thymus and in the periphery. However, susceptible ΔY-transgenic mice have elevated thymic IL-4 levels, but express almost as little IL-4 as non-transgenic NOD mice in the periphery. This drop in peripheral IL-4 production seen in ΔY-transgenic mice thus correlates with the decreased E expression in the periphery of ΔY-transgenic NOD mice. In contrast, there were no differences in IFN-γ production between the three NOD lines. We suggest that Ea-transgenic NOD mice have E-selected regulatory T cells producing IL-4, which are subsequently activated by E-expressing primary antigen-presenting cells in the periphery. This activation would then be instrumental for the E-mediated protection from disease in NOD mice. Such a process would explain the total absence of protection in ΔY-transgenic NOD mice, despite their widespread E expression.
22.	Brenden, Nina, 1967- (author) MHCI-E mediated protection from IDDM on NOD mice : The role of regulatory T cells 1998 Doctoral thesis (other academic/artistic)
23.	Cardell, Susanna, 1959- (author) T lymphocyte subsets : a study on heterogeneity based on lymphokine production 1992 Doctoral thesis (other academic/artistic)
24.	Dahlman-Höglund, Anna, 1964, et al. (author) Immune response against ovalbumin in rats colonized with an ovalbumin-producing Escherichia coli and the influence of feeding ovalbumin. 1994 In: International archives of allergy and immunology. - 1018-2438. ; 105:4, s. 381-5 Journal article (peer-reviewed)abstract The influence of feeding ovalbumin (OA) on the development of IgE/IgG antibodies and delayed-type hypersensitivity (DTH) against OA was studied in rats colonized from birth with an Escherichia coli genetically manipulated to produce OA. At 21 days of age, colonized pups and pups with a normal intestinal flora were weaned onto either an OA-containing or a conventional diet without OA. At 2 months of age the colonized rats showed an increased DTH reaction to OA, but they did not have any anti-OA antibodies in serum. The rats were then immunized intracutaneously with OA in Freund's complete adjuvant. After immunization the colonized rats fed the conventional diet had a significantly higher DTH reaction to OA and significantly higher serum levels of IgE anti-OA antibodies than the uncolonized rats on the same diet. The colonized rats eating the OA-containing diet showed a 73% decrease in the DTH reaction to OA and also significantly lower levels of IgE and IgG antibodies against OA compared with the colonized rats fed conventional diet. The dams colonized as adults by the OA-producing E. coli developed IgE anti-lipopolysaccharide antibodies in serum while the pups colonized via the dams at birth did not. Neonatal colonization with an E. coli strain producing OA resulted in increased DTH reactivity against OA and priming for secondary IgE anti-OA response. Feeding the animals an OA-containing diet from weaning abrogated this intestinally induced hypersensitivity and rendered the animals orally tolerant to OA.
25.	Elenström-Magnusson, Carina, 1961- (author) Regulation of B lymphocyte adhesion and migration by interleukin 4 1995 Doctoral thesis (other academic/artistic)
26.	Elghazali, Gehad, 1962- (author) Regulatory immune responses in humans naturally primed to Plasmodium falciparum or vaccinated with tetanum toxoid or purified protein derivative 1999 Doctoral thesis (other academic/artistic)abstract It is well established that the balance between functionally distinct regulatory CD4+ T cells plays a major role in the development of immunity and/or pathogenesis to many different infections. In spite of the importance of Th lineage commitment in disease, the critical questions how the balance between Th1 and Th2 cells is regulated is largely unresolved. This thesis describes work aimed at assessing CD4+ T cell heterogeneity and how this is regulated in humans naturally primed to P. falciparum malaria or vaccinated with tetanus toxoid or BCG.Different types of antigens have been implicated to be of importance in the polarization of Th1 or Th2 cells. To investigate this, we stimulated peripheral blood mononuclear cells (PBMC) with tetanus toxoid (TT) and the mycobacterial antigen, purified protein derivative (PPD) in vitro and determined the number of IFN-g (Th1 cytokine) and IL-4 (Th2 cytokine) producing cells using the ELISPOT assay. PPD preferentially induced IFN-g and very few IL-4 producing cells, while TT- induced both IL-4 and IFN-g. These differences probably reflect the different types of immune responses the two antigens induce, mycobacteria preferentially a cell-mediated Th1 type of immunity, while immunity to tetanus is an antibody-dependent, Th2 type of response.To investigate the role of Th1 and Th2 cells in the regulation of anti-malarial IgE in individuals living in P.falciparum endemic areas, the number of IL-4 and IFN-g producing cells was correlated to the plasma anti-malarial IgE levels. A negative correlation between the number of IFN-g producing cells and anti-P.falciparum IgE was found. For IL-4 there was a weak positive correlation with anti-malarial IgE levels, suggesting that other cells than T cells can produce IL-4. The anti-malarial IgE levels correlated significantly with an increased ratio of IL-4/IFN-g producing cells. These data suggest a regulatory role for IL-4 in the induction of anti-P.falciparum IgE antibodies.When PBMC from two groups of individuals naturally exposed to P.falciparum, living in two different parts of Africa (Burkina Faso and Tanzania), were stimulated in vitro with P.falciparum antigens, no malaria-specific IL-4 producing cells were detected. The levels of IgE were lower in the Burkina individuals as compared to the Tanzania ones. This might reflect differences in malaria exposure or genetic (ethnic) differences between the two study groups.To study the influence of genetic and/or environmental factors on the development and shaping of the human peripheral T cell repertoire, the T cell receptor (TCR) Vb usage in ten adult monozygous (Mz) and nine dizygous (Dz) twin pairs living in a P.falciparum endemic area in The Gambia was studied. The results revealed that the frequencies of cells expressing particular TCR Vb genes were not influenced by the parasitaemia, indicating that malaria exposure is not a dominating factor in shaping the peripheral TCR repertoire in humans. The mean within-pair difference was significantly lower for the Mz than for the Dz pairs. The mean within-pair difference for a group of MHC-class II identical twin pairs was significantly higher than for the Mz group but similar to that of the Dz as a whole. These data indicate that genetic factors other than MHC class II genes (i.e non-MHC) influence the shaping of the peripheral TCR Vb repertoire in humans.To study whether or not IgE-containing malaria sera have the capacity to induce IL-4 in human basophils, IgE containing sera from malaria immune donors were added to tissue culture plates coated with anti-human IgE antibodies. IgE-anti-IgE complexes induced IL-4 in basophils. Serum depleted of IgE induced significantly less IL-4. These data show that malaria IgE can induce IL-4 production in cells of basophil origin that can subsequently amplify Th2 type of responses.Taken together, our data show the existence of functionally distinct T cells in individuals naturally primed to P.falciparum or vaccinated with TT or BCG.
27.	Gontijo, Cristiano M. (author) Antigen processing and presentation by B lymphocytes decorated with passively incorporated antibodies 1994 Doctoral thesis (other academic/artistic)
28.	Goodier, M R, et al. (author) Cytokine profiles for human V gamma 9+ T cells stimulated by Plasmodium falciparum. 1995 In: Parasite immunology (Print). - 0141-9838 .- 1365-3024. ; 17:8, s. 413-23 Journal article (peer-reviewed)abstract V gamma 9+ T cells from malaria non-exposed donors make proliferative responses to Plasmodium falciparum on in vitro stimulation. V gamma 9+ cells are strongly activated by components of the schizont stage of the parasite and by antigens released into the culture upon schizogony, while CD4+V gamma 9- cells are stimulated by the earlier stages of the parasite. Using reverse transcriptase-polymerase chain reaction (RT-PCR) we determined mRNA expression for 14 cytokines in highly purified V gamma 9+ cells enriched by positive selection after in vitro stimulation with P. falciparum schizont antigens. Interferon-gamma (IFN-gamma) and Tumor Necrosis Factor-alpha (TNF-alpha) were detected in all samples tested. The majority of samples also expressed TNF-beta, transforming growth factor-beta (TGF-beta) and Interleukin-8 (IL-8). Only occasional samples expressed IL-2, IL-5 and IL-10. Using the ELISPOT assay we found that a large fraction of the reactive V gamma 9+ cells produced IFN-gamma and that gamma delta T cells are the major producers of IFN-gamma in cultures stimulated with schizont antigens. The majority of V gamma 9+ cells in these cultures also express the membrane-bound form of TNF-alpha. Expression of these cytokines speaks for a cytolytic and/or inflammatory role of gamma delta cells in the response to malaria in non-exposed individuals.
29.	Hedges, Spencer R., et al. (author) Immunoregulatory cytokines modify Escherichia coli induced uroepithelial cell IL-6 and IL-8 responses 1996 In: Cytokine. - : Elsevier BV. - 1043-4666. ; 8:9, s. 686-697 Journal article (peer-reviewed)abstract This study analysed the effects of immunoregulatory cytokines on uroepithelial cell cytokine responses. The A-498 human kidney cell line was treated with the interleukins IL-2, IL-4, IL-5, IL-10, IL-12, IL-13, interferon gamma (IFN-α) and transforming growth factor beta (TGF-β1). Secreted IL-6 and IL-8 were quantitated by enzyme-linked immunoabsorbent assay (ELISA) and bioassay; IL-6 and IL-8 mRNA species were quantitated by reverse transcriptase polymerase chain reaction (RT-PCR). IL-4, IL-13, IFN-γ, and TGF-β1, but not IL-2, IL-5, IL-10 or IL-12, stimulated IL-6 secretion. At high concentrations, IL-4 and IL-13 stimulated low levels of IL-8 secretion. Immunoregulatory cytokines were analysed for their ability to modify the A-498 cells' IL-6 and IL-8 secretion in response to Escherichia coli. IL-5, IL-12, IL-13 and TGF-β1 additively enhanced the bacterially induced IL-6 secretion, but they did not affect IL-8 secretion. The strongest affects on uroepithelial cell IL-6 and IL-8 responses in the presence of bacteria were observed in conjunction with IL-4 and IFN-α. IL-4 induced IL-6 production in synergy with E. coli. IFN-α both enhanced and inhibited IL-6 and IL-8 responses in combination with E. coli, depending on the order of stimulant addition. The results demonstrate that immunoregulatory cytokines can modify the uroepithelial cell responses to bacteria in vitro. In this way T cells may regulate the cytokine responses of uroepithelial and possibly other mucosal epithelial cells in vivo.
30.	Helmby, Helena, 1965- (author) Immune Regulation During Malaria Infection 1998 Doctoral thesis (other academic/artistic)abstract Malaria is the largest parasitic disease in the world and is caused by the protozoon Plasmodium. The most severe form in humans is caused by Plasmodium falciparum. Immunity to malaria involves both cell-mediated and humoral responses and develops slowly over a period of ten to fifteen years, requiring repeated infections.This thesis describes work aimed at investigating some of the immunological mechanisms occurring during human and experimental murine infections. The results show that polyclonal and specific Immunoglobulin (Ig) E elevations occur during blood-stage P.falciparum and P.chabaudi infections and that it is the malaria parasite itself that is responsible for this IgE induction. Elevated levels of IgE in humans are correlated with severity of disease. Studies in mice showed that the genetic background and the number of infections influence the development of IgE antibodies.Interleukin (IL)-4 plays a major role in the switching of IgM/IgG to IgE antibody production and functions as a B cell stimulatory factor. Activated T cells are a major source for IL-4 in the immune response but recently other non-lymphocyte cell types, potent producers of IL-4, have been described. One such cell type is the FceRI+ non-B non-T (NBNT) cell, probably a cell of the basophilic lineage. The data presented in this thesis show that a population of IL-3-responsive IL-4-producing NBNT cells expand in the spleens of P.chabaudi infected mice during and shortly after peak parasitemia. During this time period the mice are anemic and display elevated levels of IL-3 in their serum. The expansion of IL-4 producing NBNT cells correlates with the switch from Th1 to Th2 that takes place in the spleen during this time period. Thus, NBNT cells may represent a source of IL-4 involved in directing the Th-response towards a Th2 phenotype.The changes in cellular dynamics that take place in the spleen and peripheral blood during a malaria infection are complex, and the kinetics apparently differ between spleen and peripheral blood. The changes in the spleen involve apoptosis during the peak parasitemia. Thus, apoptosis may be of importance in the control and development of the splenic immune response.Concomitant infections with the trematode worm Schistosoma mansoni and P.chabaudi malaria resulted in higher malaria parasitemia and increased mortality. The malaria specific Th1 response was not affected by the concomitant S.mansoni infection but a significant suppression of macrophage activity was seen. The malaria infection suppressed S.mansoni-specific antibody production and Th2 cytokine responses showing that both pathogens affected the immune response to the other infection. These data emphasize the importance for similar studies in humans since multiple infections are common in malaria endemic areas.
31.	Henriksson, Elisabet Welin, 1960-, et al. (author) Autoepitope-mapping of the U1-70K protein with human-Drosophila chimeric proteins 1997 In: Journal of Autoimmunity. - : Academic Press. - 0896-8411 .- 1095-9157. ; 10:6, s. 559-568 Journal article (peer-reviewed)abstract The 70K protein is the major autoantigen for anti-RNP autoantibodies directed against the U1 small nuclear ribonucleoprotein complex particle. The U1-70K protein has been epitope-mapped by various groups, and a major antigenic region of about 70 amino acids has been found which overlaps with the RNA binding motif. Attempts to map the major antigenic region further with smaller cloned fragments or with peptides have been hampered by total loss of, or strongly reduced, antigenicity. Thus the major antigenic region is composed of conformational epitopes and a detailed analysis of particular epitopes has not been possible. In the present work, we examine the antigenicity of chimeric proteins assembled from the highly conserved Drosophila melanogaster 70K proteins grafted with human 70K segments. With this approach, the effects on antigenicity of exchanging particular segments can be assayed with the overall structure of the major antigenic domain kept relatively constant. Our results, supported by depletion experiments, show that residues 99-128 from the human protein are essential for recognition by both human and canine anti-RNP autoantibodies. These residues have to be presented in a manner that allows correct conformational interaction between the different protein domains.
32.	Henriksson, Elisabet Welin, 1960-, et al. (author) Human anti-RNP sera contain both human-specific and cross-reactive anti-70K autoantibodies 1996 In: Journal of Autoimmunity. - : Academic Press. - 0896-8411 .- 1095-9157. ; 9:4, s. 551-559 Journal article (peer-reviewed)abstract The U1 snRNP (small nuclear ribonucleoprotein complex) associated 70K protein is the main autoantigen for the anti-RNP autoantibodies which are directed against the U1 snRNP particle. The major antigenic region of the 70K protein has by various laboratories been mapped to an RNA binding domain required for the 70K-U1 snRNA interaction. We have used recombinant proteins comprising this region from the human and the Drosophila melanogaster 70K proteins to examine the species specificity of the human anti-70K autoantibodies found in 42 patient sera. Most, but not all, anti-70K positive sera in this cross-sectional sample contained both human 70K specific anti-bodies and Drosophila 70K reactive antibodies. Results of a longitudinal follow-up of 14 patients indicated that the cross-reactive anti-70K antibodies developed secondarily to the establishment of a species-specific anti-70K reaction. In a fraction of the patient sera this broadening of the response never occurred. Taken together, the data in this study support the hypothesis that the endogenous human 70K protein is the immunogen driving the production of anti-70K autoantibodies.
33.	Hu, Hui (author) Mercury-induced autoimmunity : mechanisms of susceptibility/resistance 1998 Doctoral thesis (other academic/artistic)
34.	Högstrand, K., et al. (author) Gene conversion of major histocompatibility complex genes is associated with CpG-rich regions 1999 In: Immunogenetics. - : Springer Science and Business Media LLC. - 0093-7711 .- 1432-1211. ; 49:5, s. 446-455 Journal article (peer-reviewed)abstract We examined 32 DNA sequences of mouse and human major histocompatibility complex (MHC) genes believed to have been subjected to gene conversion events. All regions of the mouse H2 genes as well as the human HLA genes which have been implied to be involved in gene conversion events had elevated levels of CpG dinucleotides, whereas the rest of the genes showed extensive CpG suppression. Mouse MHC genes which have been suspected but not directly implied to be involved in gene conversion events also showed elevated levels of CpG dinucleotides. Moreover, both mouse and human MHC genes which have never been suspected of undergoing gene conversion had low levels of CpG throughout the genes. These results indicate that high CpG levels are correlated with gene conversion rather than with polymorphism, as non-polymorphic genes that have been implicated as gene conversion donors also have elevated levels of CpG dimers in the involved regions whereas polymorphic genes which have never been considered to undergo gene conversion events have a low level of CpG dinucleotides. We also studied the methylation pattern of CpG dimers in the Abk gene by restriction enzyme digestion of mouse testis DNA followed by Southern blot and hybridization to an Abk-specific probe. The examined CpG dimers in prepubescent mice, where the latest germline stages are spermatogonia, leptene, or pachytene, are respectively non-methylated. Accordingly, the CpG dimers appear to be non-methylated in germline DNA from the testis of prepubescent mice, where gene conversions have been reported to occur.
35.	Höidén, Ingmarie, 1960- (author) Regulation of superantigen-induced cytokine production 1995 Doctoral thesis (other academic/artistic)
36.	Iqbal, Jamshaid (author) Erythrocyte membrane associated Plasmodium falciparum antigens, involvement in immunity and sequestration 1994 Doctoral thesis (other academic/artistic)
37.	Jiang, Yun, 1968- (author) Heavy Metal Superantigen : Genetic Regulation of Lymphocyte Activation by HgC12 1998 Doctoral thesis (other academic/artistic)abstract Injections of subtoxic doses of HgCl2 (mercuric chloride) cause a systemic autoimmune disease in genetically predisposed rats and mice. My work was designed to elucidate how HgCl2 induces lymphocyte activation and how the response to HgCl2 is regulated in susceptible and resistant strains.We showed that HgCl2 at a very low dose (10 mM) primarily activated murine T lymphocytes to proliferation in the presence of adherent cells in a primary in vitro culture, reaching a peak by day 5. Consistent with the in vivo findings, lymphocytes from different strains differed in their capability of responding to HgCl2. Anti-CD4 antibody completely inhibited the proliferation induced by HgCl2, which indicates that CD8+ T cells cannot be activated to division without the help from CD4+ T cells. Moreover, CD4+ T cells from susceptible A.SW and BALB/c mice, but not from resistant DBA/2 mice, were preferentially activated by HgCl2, as indicated by expression of the very early activation antigen CD69 and transformation to blast cells. Unlike CD4+ T cells, CD8+ T cells were activated by HgCl2 irrespective of the origin of the cells. Thus, helper T cells play a crucial role in the immunological effects caused by HgCl2 and may determine the ability of different strains to respond to HgCl2.We found that HgCl2 selectively activated T cells expressing certain TCR Vb elements. Depletion of Vb8+ T cells strongly suppressed HgCl2 -induced proliferation and transformation in spleen cells from BALB/c mice, indicating a specific correlation between Vb usage and the response to HgCl2. It seems unlikely that a specific TCR repertoire predisposes to the development of autoimmunity caused by HgCl2. We also verified that the unresponsiveness of CD4+ T cells from DBA/2 mice to HgCl2 was not due to immunosuppression mediated by CD8+ T cells.We further investigated the production of IL-2, IL-4, IFN-g, and IL-10 after activation by HgCl2, but found no evidence for the suggestion that HgCl2 induces a Th1/Th2 imbalance in resistant/ susceptible strains. A high frequency of IL-2-secreting cells was observed in spleen cells from the high responder A.SW mice, followed by cells from the intermediate responder BALB/c mice, whereas the frequency was low in cells from the non-responder DBA/2 mice. We showed that neutralizing anti-IL-2 antibody profoundly inhibited the proliferative response to HgCl2. Supplement of exogenous IL-2 enabled spleen cells from DBA/2 mice to respond to HgCl2 to a level comparable with cells from BALB/c mice. These findings indicate that lymphocytes from resistant mice have the potential of responding to HgCl2, but the response is limited due to an insufficient source of IL-2.HgCl2 activated B cells from susceptible mice, but not from resistant mice, to produce IgM antibody in a primary in vitro culture.We hypothesize that mercuric ions bind to certain molecules and transform them to super-antigens which in turn polyclonally activate T cells in a Vb-specific manner. This is followed by a polyclonal activation of B cells, resulting in autoantibody production and pathological manifestations in susceptible strains. IL-2 may be a limiting factor that precludes lymphocytes from resistant strains from responding to HgCl2.
38.	Johansson, S, et al. (author) Towards studies of interactions between echovirus 7 and the receptor on human cells: sequence analysis of an infectious full-length clone 1997 In: 12th International Symposium on Affinity Interactions: Fundamentals and Applications of Biomolecular Recognition, Kalmar, Sweden. Conference paper (peer-reviewed)
39.	Kabilan, Lalitha (author) T-cell regulation of immune responses to the Plasmodium falciparum antigen Pf155/RESA 1990 Doctoral thesis (other academic/artistic)abstract Plasmodium falciparum is the cause of the most severe malaria inhumans. Due to the rapid resurgence of malaria in many parts of theworld extensive efforts have been made during the last few years todefine several P.falciparum antigens for inclusion in a vaccine. Onesuch antigen, generally considered to be a candidate for inclusion in asubunit vaccine against the asexual blood stages of the parasite isPfl55/RESA. Several antibody-binding structures (B-cell epitopes) ofthe molecule have recently been identified. However, immunogens for asubunit (malaria) vaccine should also contain T-cell activating sitesderived from the parasite in order to induce good secondary responsesafter natural boosting.For this reason, we have investigated Pfl55/RESA for its T-cellactivating capacity. Here we have shown, in Pfl55/RESA seropositivedonors, that the antigen induced production of anti-Pfl55/RESAantibodies in vitro is T-cell dependent, implying that Pfl55/RESAcontains T-helper cell stimulating epitopes. We have also shown thatsynthetic peptides representing sequences from the amino-acid repeatregions of Pfl55/RESA stimulate T-cells from P.falciparum-primed donorsto proliferate, to release IFN-7 and/or to synthesize IL-4. The resultssuggest the presence of several T-cell epitopes in the conserved repeatregions of Pfl55/RESA. In individual donors, there was no correlationbetween these different activities. Rather, they were negativelyassociated, suggesting that they have exhibited by functionallydistinct T-cells perhaps similar to the TH1 and TH2 cells found inmice. The results also demonstrate the importance of examining multipleparameters of T-cell activation when estimating the proportion ofindividuals responding to any particular epitope. However, induction ofIL-4 was seen in donors who had elevated concentrations of serumantibodies to the peptide used for T-cell activation. These resultssuggest that IL-4 producing cells have a role in the induction of theformation of Pfl55/RESA specific antibodies.To measure the actual number of cells secreting IFN-7 in vitro inresponse to Pfl55/RESA peptides, a single cell assay (ELISPOT) wasdeveloped. The results show that small numbers of antigen specificIFN-7 producing cells could be detected by ELISPOT even in the absenceof detectable levels of IFN-7 secreted into the culture supernatant.Thus, the Elispot makes it possible to estimate the frequency of Tcellsproducing different lymphokines even when cell numbers arelimited. As it requires only small volumes of blood and is relativelysimple, it is a useful assay for large scale epidemiological studies ofT-cell responsiveness in malaria or other diseases.
40.	Koho, Hannu, 1953- (author) Antigens associated with human urinary bladder carcinoma : identification and functional studies 1990 Doctoral thesis (other academic/artistic)abstract The primary aim of this study was to identify antigens associated with themalignant state of human urinary bladder carcinoma cells. Characterization ofsuch tumor-associated antigens should provide clues to the molecularmechanisms of tumorigenesis, as well as leading to possible clinical applications,such as immunodiagnosis and immunotherapy.Five novel cell-surface glycoproteins were identified by monoclonalantibodies raised against bladder carcinoma cells. Four of these were selectivelyexpressed by malignant urothelial cells. One of the antigens was also identified asa differentiation antigen on B-lymphocytes, and has subsequently beendesignated CD40 in accordance with international nomenclature. This andanother antigen, 8F4, are putative growth factor receptors. Functional studiesshowed that these two antigens, CD40 and 8F4, are phosphoproteins. The 8F4antigen also displayed protein kinase activity, similar to many growth factorreceptors and receptor-like oncogene products. The CD40 antigen has anestablished role in the growth regulation of B-lymphocytes, as demonstrated bythe capacity of anti-CD40 antibodies to stimulate growth of preactivated Blymphocytes.Molecular cloning has also identified the antigen as a putativegrowth factor receptor related to the NGF-receptor. Structural analysis of theCD40 gene in bladder carcinoma cell lines by cDNA cloning and DNA sequencinganalysis showed no differences from the CD40 gene expressed in B cells. Theover-expression of CD40 in bladder carcinomas is not likely associated withstructural changes in the transcribed portion of the gene. Possible reasons for theover-expression are discussed. Expression of antigens was affected by interferongammatreatment, in that three antigens (CD40, 8F4, and 4E8) were up-regulatedand one (2F6) was down-regulated. Although the identified antigens areassociated with malignancy, their role in the transformation process remains tobe investigated. In this respect, molecular cloning is an important tool forfunctional characterization of the other antigens and should also reveal whetherthere are additional receptors among them. It will also be of interest to examinethe possible involvement of these antigens in tumor invasion and metastasis.
41.	Kulane, Asli (author) In Vitro Studies of Factors Potentially Affecting Plasmodium Falciparum Infection : (Heparin and Anti-P. falciparum Immune Responses) 1997 Doctoral thesis (other academic/artistic)abstract Plasmodium falciparum malaria is considered one of the major infectious diseases in humans, with regard to mortality and morbidity. Growing resistance of malaria to most anti-malaria drugs and of the Anopheles mosquitoes to insecticides have resulted in a global resurgence of the disease. Therefore, the need to explore drugs with possible antimalarial effects or the development of vaccines against malaria are considered a high priority for control of the disease. In this thesis, the antimalarial effect of heparin and the identification of T- and B-cell epitopes in P. falciparum vaccine candidate antigens, Pf155/RESA and Pf332 have been addressed.Heparin, a drug included in the supportive or ancillary treatment of cerebral malaria, was tested for its anti-parasitic effect on the blood stages of P. falciparum malaria in vitro. Heparin was cleaved into fragments differing in molecular weight and in their affinity for antithrombin III. Both unfractionated heparin and heparin fractions inhibited the merozoite invasion into red blood cells. The mechanism by which heparin acts is not clear. However, the inhibition was reversible by washing heparin-treated P. falciparum cultures indicating a direct effect of heparin. The sensitivity of laboratory strains and/or fresh isolates obtained from individuals residing in malaria endemic areas was compared. Although varying in sensitivity none of the samples tested was found to be resistant to heparin and its derivates. A fraction of heparin with low affinity for antithrombin III was the most potent inhibitor of merozoite invasion. The fraction with low affinity for antithrombin III is devoid of anticoagulant activity, suggesting a potential role of this fraction for the treatment of malaria.For the development of subunit vaccines it is important to identify and characterise epitopes which activate relevant B- and T-cell functions. In this study we have investigated two putative malaria vaccine candidate antigens, Pf155/RESA and Pf332, for T- and B-cell reactive epitopes. A large number of donors had antibodies against the Pf155/RESA sequence 186-206. For the Pf332 derived fragment, EB200, the donors also exhibited high antibody levels in their plasma. In our studies we have measured multi parameters of T-cell activation (proliferation, IFN-g and/or IL-4 production). Peptides corresponding to the N-terminal region of Pf155 or to EB200, stimulated peripheral blood mononuclear cells from P. falciparum-immune donors to proliferate, to induce secretion of IFN-g and/or IL-4. In individual donors the cellular immune responses to the peptides varied considerably. However, there was no clear association between proliferation and production of the cytokines investigated. This lack of association underlines the importance of including multiple parameters when analysing T-cell responses to defined epitopes.In conclusion, the non-repeat region of Pf155/RESA and the EB200 fragment of Pf332 contains several B-cell epitopes as well as several epitopes inducing functionally distinct T-cell responses, which should be a useful tool for inclusion in a subunit malaria vaccine as well as in future immuno-epidemiological studies.
42.	Kullberg, Marika, 1965- (author) Immunoregulation and cytokine production during Schistosoma mansoni infection 1995 Doctoral thesis (other academic/artistic)
43.	Leijon, Kristina, 1967-, et al. (author) Specific destruction of islet transplants in NOD‹–›C57BL/6 and NOD‹–›C3H/Tif embryo aggregation chimeras irrespective of allelic differences in beta-cell antigens 1995 In: Journal of Autoimmunity. - : Elsevier. - 0896-8411 .- 1095-9157. ; 8:3, s. 347-356 Journal article (peer-reviewed)abstract We have tested the hypothesis that allelic differences in the antigens expressed by the beta-cells of the islets of Langerhans influence the development of insulitis in the non-obese diabetic (NOD) mouse. Islets of Langerhans from NOD, C57BL/6 and C3H/Tif mice were transplanted under the kidney capsule of NOD<-->C57BL/6 and NOD<-->C3H/Tif embryo aggregation (EA) chimeras and the infiltration was scored 5-7 weeks later. Mononuclear cell infiltration of pancreatic islets was observed in 60% of the NOD<-->C57BL/6 and in 55% of the NOD<-->C3H/Tif EA chimeras. All transplanted EA chimeras that developed insulitis also displayed mononuclear cell infiltrates in the transplants, irrespective of the origin of the transplanted islets. In contrast, no infiltration of transplants was detected in EA chimeras scoring negative for insulitis. These results demonstrate that the specific destruction of islet transplants does not require the expression of NOD specific antigens by the islets. Moreover, the beta-cell destruction appears not to be restricted to NOD-MHC. The correlation between insulitis and transplant beta-cell destruction suggests the possibility that the development of insulitis is a prerequisite for transplant specific destruction. MHC restricted destruction may, therefore, precede the beta-cell destruction of transplanted islets. The chimerism among the mononuclear cells infiltrating the islet transplants was found to correlate with the overall haematopoetic chimerism in each of the individual EA chimeras. This observation suggests that NOD bone marrow, as well as non-NOD bone marrow, generates cells contributing to the beta-cell destruction process.
44.	Lindberg, Johan, 1960- (author) Structural studies of bacterial polysaccharides using N.M.R. spectroscopy and mass spectrometry 1990 Doctoral thesis (other academic/artistic)abstract This work describes the structural studies of the capsularpolysaccharides from Streptococcus pneumoniae type 18F and 7B,Alcaligenes ATCC 31961 (S-194), and the O-antigen polysaccharidefrom Vibrio cholerae 0:3.The principal methods used in these studies were ID and 2D n.m.r.spectroscopy, e.i. and f.a.b. mass spectrometry, and specific chemicaldegradations.
45.	Los, Marek Jan, et al. (author) Human T cell leukemia virus-I (HTLV-I) Tax-mediated apoptosis in activated T cells requires an enhanced intracellular prooxidant state 1998 In: Journal of Immunology. - : American Association of Immunologists. - 0022-1767 .- 1550-6606. ; 161:6, s. 3050-3055 Journal article (peer-reviewed)abstract We have shown that an estradiol-dependent activation of human T cell leukemia virus-I Tax leads to the inhibition of cell proliferation and to the induction of apoptosis, The present study demonstrates that a hormone-dependent activation of Tax promotes an enhanced prooxidant state in stably transfected Jurkat cells as measured by changes in the intracellular levels of glutathione and H2O2; these changes are followed by apoptotic cell death. Additional stimulation of the CD3/TCR pathway enhances the oxidative and apoptotic effects. Both Tax-mediated apoptosis and oxidative stress can be potently suppressed by antioxidants, as is seen with the administration of recombinant thioredoxin (adult T cell leukemia derived factor) or pyrrolidine dithiocarbamate. Hormone-induced Tax activation induces a long-lasting activation of NF-kappa B, which is a major target of reactive oxygen ntermediates. The long-term exposure of Jurkat cells to hormone eventually results in a selection of cell clones that have lost Tax activity. A subsequent transfection of these apparently "nonresponsive" clones allows the recovery of Tax responses in these cells. Our observations indicate that changes in the intracellular redox status mag be a determining factor in Tax-mediated DNA damage, apoptosis, and selection against the long-term expression of Tax function.
46.	Lundqvist, Carina, et al. (author) Cytokine profile and ultrastructure of intraepithelial gamma delta T cells in chronically inflamed human gingiva suggest a cytotoxic effector function 1994 In: Journal of Immunology. - 0022-1767 .- 1550-6606. ; 153:5, s. 2302-2312 Journal article (peer-reviewed)abstract We have shown that gamma delta T cells in human gingiva have an intraepithelial location and, that in the chronic inflammatory disease periodontitis, the expression of CD45RO and CD8 or CD4 is induced on gamma delta T cells. To study the role of gamma delta T cells in local antibacterial responses, we determined the cytokine profiles of isolated human gingival cells. Different T cell subpopulations, isolated by positive selection with mAb-coated magnetic beads and macrophages, as well as epithelial cells, were analyzed for expression of mRNA for 15 cytokines by reverse transcriptase-PCR. The ultrastructure of gingival gamma delta T cells was also studied. The gamma delta T cells expressed mRNA for IFN-gamma, TNF-alpha, TGF-beta 1, and IL-6. Expression of IFN-gamma was a consequence of inflammation. CD4+ gamma delta T cells expressed IFN-gamma only, whereas CD8+ gamma delta T cells expressed all four cytokines. CD8+ cells expressing IFN-gamma, TNF-alpha, and IL-6 in combination suggest a cytotoxic effector function. Gingival gamma delta T cells contained cytoplasmic electron-dense membrane-bound granules and multivesicular bodies that are ultrastructural characteristics of cytotoxic cells. Epithelial cells from inflamed gingiva expressed HLA-DR, CD1a, CD1c, and heat shock protein 60 on the cell surface. They also expressed mRNA for IL-1 beta, IL-6, IL-8, TNF-alpha, and TGF-beta 1. Thus, epithelial cells may function as accessory cells in immune activation and, at the same time, be target cells for CD8+ gamma delta T cells reactive with CD1 Ag or heat shock protein. These results suggest that gamma delta T cells constitute a first line of defense in gingiva, preventing entrance of pathogens by cytotoxicity against infected and stressed epithelial cells, and by control of epithelial cell growth through secretion of regulatory cytokines.
47.	Lundqvist, Carina, et al. (author) Intra-epithelial lymphocytes. Evidence for regional specialization and extrathymic T cell maturation in the human gut epithelium 1995 In: International Immunology. - : Oxford University Press (OUP). - 0953-8178 .- 1460-2377. ; 7:9, s. 1473-1487 Journal article (peer-reviewed)abstract The human gut epithelium is a unique immunological compartment, containing substantial amounts of intra-epithelial lymphocytes (IEL) with unknown functions. In this study we show that distinct and unusual subpopulations of IEL are present at different levels of human intestine. IEL phenotypes in normal jejunum, ileum and colon were compared using immunoflow cytometry and immunohistochemistry. The expression of mRNA for recombination-activating gene-1 (RAG-1) in IEL from all three levels was compared using reverse-transcription polymerase chain reaction, and the morphology of IEL in situ was determined using immunoelectron microscopy. Surface marker profiles of isolated intestinal epithelial cells at all three levels were also investigated. On average the proportion of TCR gamma delta IEL was comparable in jejunum than ileum and colon and varied in phenotype with gut level. CD4-CD8-TCR alpha beta IEL dominated in colon but were absent in jejunum. CD8+ TCR alpha beta IEL were present at all levels but only in jejunum did they constitute the majority of all IEL. CD4+ TCR alpha beta IEL were present in similar frequencies at all levels of the gut. In general, the majority of IEL had an activated phenotype (CD45RO+, alpha E beta 7+). Furthermore, IEL exhibited phenotypes which are rare in peripheral blood. The thymocyte markers CD1a and CD1c as well as the NK cell marker CD56 were expressed on a fraction of TCR alpha beta and TCR gamma delta IEL. A small population of 'null' cells (CD45+ TCR/CD#-CD20-CD14-CD15- cells) was also present at equal proportions along the gut. Jejunal but not colonic IEL expressed RAG-1 mRNA suggesting that extrathymic T cell maturation occurs in the epithelium of small intestine. RAG-1 was expressed in CD2+TCR/CD3- and CD3+/TCR-IEL. Ultrastructurally, IEL often formed small clusters and intimate contacts with epithelial cells, suggesting cell cooperation within the epithelium. Some IEL had pseudopodium-like extensions penetrating the epithelial basement membrane suggesting transmigration. Epithelial cells in small intestine but not colon expressed heat shock protein 60 and HLA-DR. CD1a, CD1b and CD1c were not expressed on intestinal epithelial cells at any level. The distinct surface marker profiles of IEL and epithelial cells along small and large intestine suggest functional regional specialization and are compatible with the hypothesis that TCR alpha beta IEL participate in immune reactions to lumenal antigens while TCR gamma delta IEL perform surveillance of the epithelium.
48.	Lundqvist, C, et al. (author) Intraepithelial lymphocytes in human gut have lytic potential and a cytokine profile that suggest T helper 1 and cytotoxic functions. 1996 In: Journal of Immunology. - 0022-1767 .- 1550-6606. ; 157:5, s. 1926-34 Journal article (peer-reviewed)abstract The functional properties of intraepithelial lymphocytes (IEL) in normal human jejunum, ileum, and colon were investigated. Cytokine mRNA expression in IEL and enterocytes was determined by reverse transcriptase-PCR and IFN-gamma+ IEL by immunohistochemistry. Polyclonal activators were used to study proliferation and IFN-gamma secretion of IEL, and an anti-CD3-mediated redirected cytotoxicity assay was used to determine the lytic potential of IEL. Freshly isolated IEL at all three gut levels expressed mRNA for IL-1 beta, IL-2, IL-8, IFN-gamma, and TNF-alpha. Approximately 10% of IEL produced IFN-gamma, suggesting that IEL are immunologically active in vivo, performing similar functions along the intestine. IEL could be stimulated further in vitro to express IL-10, TNF-beta, and TGF-beta 1, while no Th2-type cytokines were induced, suggesting suppressive and cytolytic functions for IEL. All three jejunal IEL subpopulations (CD4-CD8-TCR-gamma delta+, CD4+TCR-alpha beta+, CD8+TCR-alpha beta+) expressed the same four cytokines, IL-2, IL-8, IFN-gamma, and TNF-alpha, indicating that CD4+TCR-alpha beta+ IEL are Th1 cells and that TCR-gamma delta+ IEL and CD8+TCR-alpha beta+ IEL include cytotoxic effector cells. Indeed, freshly isolated jejunal IEL displayed cytolytic activity. IEL were induced to proliferation by anti-CD3/TCR complex mAbs and leukoagglutinin, but not by Con A. There was no correlation between the magnitude of the proliferative response and the amounts of secreted IFN-gamma. Enterocytes expressed IL-1 beta and IL-8, and sometimes TNF-alpha. Although jejunal enterocytes express HLA-DR and hsp60, Ag presentation by these cells may induce anergy since their cytokine profile is different from that of classical APCs.
49.	Lundqvist, Carina, et al. (author) T-cell receptor gamma delta-expressing intraepithelial lymphocytes are present in normal and chronically inflamed human gingiva 1993 In: Immunology. - 0019-2805 .- 1365-2567. ; 79:1, s. 38-45 Journal article (peer-reviewed)abstract The phenotypic profile of leucocytes in diseased and normal gingival tissue was studied in situ and in isolated gingival mononuclear cell (GMC) preparations. T-cell receptor (TcR)gamma delta + cells showed preferential localization to epithelium, both in normal and inflamed gingiva, and were present in crevicular as well as oral epithelium. In normal gingiva > or = 30% of the isolated leucocytes expressed TcR gamma delta, of which the majority were CD4- CD8-, and expressed CD45RA. The proportion of TcR gamma delta + cells in GMC from periodontitis tissue varied between 2 and 32%. In contrast to normal gingiva the majority of TcR gamma delta + cells in diseased tissue were CD8+ and expressed CD45RO. Thus expression of the CD8 antigen on gingival TcR gamma delta + cells is probably a consequence of immune activation. Numerous Langerhans' cells and keratinocytes expressing the major histocompatibility complex (MHC) class I-like antigen, CD1, were present within normal and inflamed gingival epithelium in close proximity to the TcR gamma delta + cells. Most CD1a+ cells were scattered within oral epithelium. CD1c+ cells were localized close to the basal layer of crevicular epithelium. No CD1b+ cells were found. TcR alpha beta + cells, CD4+ and B cells were restricted to lamina propria of periodontitis lesions. The presence of intraepithelial TcR gamma delta + cells in normal gingiva suggests that they constitute the 'first line of defence' against the potentially harmful microflora in the oral cavity. Induction of CD8 and CD45RO antigens on TcR gamma delta + cells in periodontitis tissue indicate that they play a significant role in the disease. CD1 molecules on Langerhans' cells and keratinocytes may be the restriction elements for the CD8+ TcR gamma delta + cells.
50.	Mattsby-Baltzer, Inger, 1949, et al. (author) IL-1beta, IL-6, TNFalpha, fetal fibronectin, and endotoxin in the lower genital tract of pregnant women with bacterial vaginosis. 1998 In: Acta obstetricia et gynecologica Scandinavica. - : Wiley. - 0001-6349. ; 77:7, s. 701-6 Journal article (peer-reviewed)abstract In our studies on women with bacterial vaginosis (BV) in early pregnancy a strong association has been found between BV and the levels of endotoxin or interleukin-1alpha (IL-1alpha) in the lower genital tract. In the present study we investigated if an association could be found between BV and other cytokines (IL-1beta, IL-6, tumor necrosis factor alpha, TNF) or fetal fibronectin (FFN). The cytokine-inducing capacity of endotoxins present in the cervical mucus was explored in a monocytic cell assay.Cervical mucus or cervicovaginal fluid was collected from women with (BV) and without BV (nonBV) attending a family planning unit for first trimester abortion. The concentrations of IL-1beta, IL-6, TNF and FFN were determined by quantitative enzyme immunoassays. TNF was determined in 63 women (BV, n=25) out of whom 37 (BV, n=11) were analyzed for IL-1beta and the remaining 26 for IL-6 (BV, n=14). FFN was determined in another 36 women (BV, n= 19). The cytokine-inducing capacity of endotoxin-containing cervical mucus and purified endotoxin of Prevotella bivia were studied by an in vitro cell assay using a human monocytic cell line (THP-1).IL-lbeta and IL-6 were found in almost all women. The levels of IL-1beta, but not IL-6, TNF or FFN, were significantly increased in women with BV compared with the nonBV women (p<0.05). Purified endotoxin from P. bivia, and cervical mucus from BV women containing high levels of endotoxin were able to induce a cytokine response (IL-6) in monocytic cells in vitro.BV is associated with increased levels of IL-1beta in the lower genital tract of pregnant women in the first trimester. The ability of BV-associated endotoxins to induce cytokine production in monocytic cells may partly explain the increased IL-1beta levels.

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