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Biomedical subjects

K Gustafsson

Publications and source records attributed to K Gustafsson.

At least 55 records · Page 3Linked to original sources
PubMed1997

Suppression of MHC class II expression by human class II trans-activator constructs lacking the N-terminal domain.

The class II trans-activator (CIITA) is a bi- or multi-functional domain protein which plays a critical role in the expression of MHC class II genes. We report that removal of the N-terminal 151 amino acids, encompassing all of the acidic domain but leaving intact the proline/serine/threonine-rich domain, results in a mutant protein with potent suppressive properties for MHC class II expression. HeLa cells stably or transiently transfected with mutant CIITA constructs showed up to 99% suppression of MHC class II antigen induction by IFN-gamma and marked suppression of HLA-DRA mRNA expression. Transient transfection of a B lymphoma line resulted in up to 89% reduction of constitutive MHC class II expression within 5 days and suppression of HLA-DRA mRNA synthesis.

B-Lymphocytes↗
PubMed1997

Meningoencephalitis in capercaillie (Tetrao urogallus L.) caused by a Sarcocystis-like organism.

A nonsuppurative meningoencephalitis, previously presumed to be toxoplasmosis, was found in 53 capercaillies (Tetrao urogallus L.) examined at necropsy at the National Veterinary Institute, Uppsala, Sweden, between 1966 and 1985. Pronounced meningitis and encephalitis with perivascular cuffs of mononuclear inflammatory cells as well as focal gliosis were prominent histopathologic findings. Protozoa were frequently associated with these lesions. Ultrastructurally, the protozoa appeared to divide by endopolygeny, and merozoites had no rhoptries. Organisms from all 12 birds subjected to Sarcocystis cruzi immunohistochemical staining reacted positively but did not react to Toxoplasma gondii antiserum. The agent was, therefore, assigned to the family Sarcocystidae and was probably more closely related to species of the genus Sarcocystis than to T. gondii.

Animals↗
PubMed1996

Attempts to demonstrate indirect T cell allorecognition of donor MHC peptides in transplant patients.

Indirect T cell allorecognition has been shown to play an important role in the rejection of allografts in experimental animals. Although there has been much speculation as to its role in clinical transplantation, especially with regard to chronic rejection, indirect T cell allorecognition has been difficult to demonstrate in transplant patients. In this paper, we looked for in vitro T cell proliferation to synthetic peptides corresponding to donor HLA-A and HLA-B incompatible antigens. Twelve 15 amino acid peptides corresponding to the hypervariable regions of six of the most common HLA class I alleles in Caucasian populations (A1, A2, A3, B7, B8 and B44) were studied. Blood was taken from 12 adult patients following one or more episodes of acute kidney graft rejection, and from three pediatric patients undergoing chronic rejection of heart/lung transplants. The donor-recipient combinations were selected such that at least one of the six HLA antigens above was present in the donor and absent in the recipient. Peripheral blood mononuclear cells from these patients responded strongly in proliferation assays to phytohemagglutinin. However, none responded to the incompatible donor HLA peptides. Compartmentalization of responding T cells, the effects of immunosuppression, and assay sensitivity are discussed as possible explanations for the negative results.

Adult↗
PubMed1996

Pig alpha1, 3galactosyltransferase: a major target for genetic manipulation in xenotransplantation.

Terminal carbohydrate residues of glycolipids and glycoproteins display polymorphism among as well as within various species. With the exception of Old World monkeys, great apes and man, the Gala1,3Gal structure is widely expressed in all mammals examined so far. The lack of expression of the glycosyltransferase responsible for the synthesis of Gala1,3Gal leads to the production of high titers of natural antibodies (NAb) against the Gala1,3Gal of other species. The inactivation of this gene occurred during early evolution of primates. Neutralization of viruses (e.g. retroviruses) carrying the epitope, by the pre-formed human NAb, indicates one possible evolutionary reason for the polymorphism of terminal carbohydrates among as well as within species. It has been shown that this epitope constitutes the major target, on pig endothelial cells (EC), for the pre-formed human NAb resulting in a hyperacute rejection (HAR) response. This currently makes transplantation of e.g. pig organs to humans impossible. Efforts are currently underway to prevent or to eradicate the expression of this epitope in transgenic pigs. Such pigs are likely to display a greatly increased resistance to the HAR.

Animals↗
PubMed1996

Sensitization of cells and retroviruses to human serum by (alpha 1-3) galactosyltransferase.

Mammalian C-type retroviruses are inactivated by human serum, following triggering of the classical complement cascade. This may have inhibited transmission to humans of C-type oncoviruses from other mammals. Indeed, the retroviruses human immunodeficiency virus and human T-cell leukaemia virus are resistant to human complement. Antibody-independent activation of human C1q, the first component of the classical pathway, by retroviral envelope proteins has been described. However, retroviruses produced from human cells are resistant to inactivation by human complement and human serum is known to contain antibodies directed against carbohydrates on retroviral envelopes. Gal(alpha 1-3)Gal terminal carbohydrates are expressed by most mammals but are absent in humans, which lack a functional (alpha 1-3)galactosyltransferase gene. Here, we demonstrate that anti-Gal(alpha 1-3)Gal antibodies in human serum inactivate retroviruses produced from animal cells. Expression of porcine (alpha 1-3)galactosyltransferase in human cells renders the cells and the retroviruses they produce sensitive to human serum.

3T3 Cells↗
PubMed1996

Structure and organization of pig MHC class II DRB genes: evidence for genetic exchange between loci.

The pig major histocompatibility complex DRB genes were studied by polymerase chain reaction (PCR) amplification of exon 2 from eight domestic pigs and two European wild boars. Sequence comparisons together with a phylogenetic analysis showed the existence of at least three DRB genes of which only one appears to be expressed. The two putative DRB pseudogenes contained deletions in exon 2, making it possible to confirm the presence of three non-allelic DRB genes by analyzing the length polymorphism of the amplified PCR products. The expressed gene shows allelic polymorphism at the same positions as in the human DRB1 gene. In addition, this pig gene shows extensive allelic polymorphism at positions 84-88, whereas, e.g., human DRB genes do not. Surprisingly, the two putative DRB pseudogenes also display a considerable amount of allelic polymorphism, albeit of a different character as compared with the expressed DRB gene. Short stretches of sequences are shared between individual alleles at different loci. These sequence similarities cannot be due to natural selection, since two of the three DRB genes involved are polymorphic pseudogenes constituting allelic series that have diverged after the inactivation event. Instead, the results indicate that the sequences have been exchanged between the DRB genes by intergenic recombination.

Animals↗
PubMed1996

Antifection: an antibody-mediated method to introduce genes into lymphoid cells in vitro and in vivo.

We have developed a simple, safe and versatile method, termed antifection, by which antibodies are used as delivery vehicles to introduce genes into cells expressing specific surface antigens. Antibodies directed against CD3, CD34 or surface immunoglobulins were covalently coupled to plasmids containing marker genes (neoR, beta-galactosidase). Such conjugates were used in vitro and/or in vivo to antifect (transfect using antifection) cells bearing the respective targeted epitope on either normal splenic B lymphocytes or lymphoid-related cell lines. In these conditions the expression of the protein encoded by the marker gene was readily detected. Antifection is a method of delivering genes through a physiological cellular pathway, receptor-mediated endocytosis, into specific cell types, and thus may be considered as an alternative for gene therapy strategy.

Antibodies↗
PubMed1996

Experimental Toxoplasma gondii infection leading to fatal enteritis in reindeer (Rangifer tarandus).

Two, 50-60-kg yearling reindeer (Rangifer tarandus) were inoculated intraruminally with 5,000 (reindeer 1) or 50,000 (reindeer 2) oocysts of the ME-49 strain of Toxoplasma gondii. Both reindeer became ill day 4 postinoculation (p.i.). Reindeer 2 died because of acute enteritis day 9 p.i. Histologically, extensive necrosis and destruction of the intestinal mucosa were seen. Numerous T. gondii organisms were demonstrated immunohistochemically. Reindeer 1 was treated with sulfatrimethoprim for 2 days from day 9, recovered by day 16 p.i., and remained clinically normal until the last day of observation (day 707 p.i.). It developed high antibody titers to T. gondii between days 7 and 14 p.i.

Animals↗
PubMed1995

Luminometry and PCR-based monitoring of gene-tagged cyanobacteria in Baltic Sea microcosms.

The cyanobacterium Synechocystis 6803 was tagged by chromosomal integration of the firefly luciferase gene, luc, resulting in the modified strain Synechocystis 6803-luc. The tagged cells were monitored in Baltic Sea microcosms both by detection of the luc gene by PCR amplification and by measurement of luc gene expression (bioluminescence) in total protein extracted from sediment and water. A new method was developed for isolation and concentration of total protein from sediment for optimization of luciferase quantitation. The detection limit for Synechocystis with a chromosomal luc insertion by bioluminescence was in the order of 4 x 10(3) cells per g sediment, a considerable improvement in sensitivity over previous methods. Another improvement was to use an internal luciferase standard to correct for quenching of light output by impurities in the samples. Baltic sea microcosms were inoculated with Synechocystis 6803-luc, and the luc DNA and luciferase protein specific to the tagged cells were monitored over time. A decrease in luminescence in the microcosm water was observed, simultaneously with an increase in luminescence in the sediment, suggesting settling of the luc-tagged cells in the sediment layer.

Base Sequence↗
PubMed1995

cDNA sequence and chromosome localization of pig alpha 1,3 galactosyltransferase.

Human serum contains natural antibodies (NAb), which can bind to endothelial cell surface antigens of other mammals. This is believed to be the major initiating event in the process of hyperacute rejection of pig to primate xenografts. Recent work has implicated galactosyl alpha 1,3 galactosyl beta 1,4 N-acetyl-glucosaminyl carbohydrate epitopes, on the surface of pig endothelial cells, as a major target of human natural antibodies. This epitope is made by a specific galactosyltransferase (alpha 1,3 GT) present in pigs but not in higher primates. We have now cloned and sequenced a full-length pig alpha 1,3 GT cDNA. The predicted 371 amino acid protein sequence shares 85% and 76% identity with previously characterized cattle and mouse alpha 1,3 GT protein sequences, respectively. By using fluorescence and isotopic in situ hybridization, the GGTA1 gene was mapped to the region q2.10-q2.11 of pig chromosome 1, providing further evidence of homology between the subterminal region of pig chromosome 1q and human chromosome 9q, which harbors the locus encoding the AB0 blood group system as well as a human pseudogene homologous to the pig GGTA1 gene.

Amino Acid Sequence↗
PubMed1995

Influence of processing and cooking of carrots in mixed meals on satiety, glucose and hormonal response.

The influence of processing and cooking on the metabolic response to carrots in mixed meals was explored in two consecutive harvest years. The contribution of dietary fibre (4.4 g 1989 and 6.6 g 1990) from carrots was chosen to be different in order to compare effects with varying doses. The meals, composed of carrots, creamed potatoes, meat balls, lingonberry jam, white bread and light beer, were served in the morning after an overnight fast to 10 healthy male volunteers. Carrots were investigated raw, processed (blanched and frozen) and variously cooked (thawed, boiled and microwaved). The amount of dietary fibre from the vegetable, and the content of energy, digestible carbohydrates, fat and protein were similar in the meals compared. Significantly lower glucose, insulin and C-peptide responses and higher satiety scores were elicited with raw carrots than with microwaved ones, harvest year 1989. The next year, with a higher dietary fibre intake from carrots, there were significant effects of processing only on the glucose response. Plasma beta-carotene levels tended to be higher postprandially with raw carrots than with microwaved ones. Hence, ordinary processing and cooking of vegetables can affect the metabolic response to a mixed meal. However, the influence seems to be varying and of minor importance in ordinary meals. Increasing vegetable portions entailing a higher soluble fibre content and a higher viscosity could further reduce the influence of processing.

Adult↗
PubMed1995

Satiety effects of spinach in mixed meals: comparison with other vegetables.

The effects of vegetables in mixed meals on satiety were evaluated by varying the dose (portions 150 and 250 g containing 4.3 and 7.2 g of dietary fibre, respectively) and structure (cut and minced) of microwaved spinach. A control meal without spinach and the test meals with spinach were balanced regarding energy (2000 kJ) and digestible carbohydrates (59 g, 51 E%) and with protein and fat as far as possible similar. The meals were served in the morning, after an overnight fast, to ten healthy male volunteers. Satiety was registered and blood glucose as well as plasma insulin and C-peptide were analysed at regular intervals until 3.5 h postprandially. The largest spinach portions augmented satiety and reduced the postprandial glucose response. The total satiety scores seemed to be correlated positively to both the dietary fibre and the water content in the vegetable. Differences in structure had no influence on satiety scores.

Adult↗
PubMed1994

Remoxipride shows low propensity to block functional striatal dopamine D2 receptors in vitro.

The effect of remoxipride ((S)(-)3-bromo-N-[(1-ethyl-2-pyrrolidinyl)methyl]-2,6-dimethoxybenzam ide) on dopamine D2 receptor-mediated inhibition of cAMP formation in rat striatal tissues pieces was established together with that of a number of other dopamine D2 receptor antagonists. The action of remoxipride, three other substituted benzamides, (-)-sulpiride, raclopride and NCQ 298 ((S)-3-iodo-N-[(1-ethyl-2-pyrrolidinyl)methyl]-5,6-dimethoxysalicylam ide mesylate) and haloperidol, a butyrophenone, was studied in the presence of (I) (+/-) SKF 38393 (7,8-dihydroxy-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine) hydrochloride (100 microM) plus pergolide (1 microM) or (II) forskolin (1 microM) plus dopamine (100 microM). In addition, four of the metabolites of remoxipride: FLA 797 ((S)-3-bromo-N[(1-ethyl-2-pyrrolidinyl)methyl]-2-hydroxy-6- methoxybenzamide), NCR 181 ((S)(-)-5-bromo-N-[(1-ethyl-2-pyrrolidinyl)methyl]-2-hydroxy-6- methoxybenzamide tartrate), NCQ 436 ((S)(-)-3-bromo-N-[(1-ethyl-2-pyrrolidinyl)methyl]-5,6-dihydroxy-2- methoxybenzamide semioxalate) and NCQ 469 ((S)(-)-3-bromo-N-[(1-ethyl-2-pyrrolidinyl)methyl]-5-hydroxy-2,6- dimethoxybenzamide hydrochloride), mainly found in rodents, were studied using test system I. The results demonstrate that remoxipride is significantly weaker in blocking functional striatal dopamine D2 receptors than either of the reference compounds studied and three of the four metabolites. The studies also demonstrate that dopamine D1 and D2 receptor interactions at the level of cAMP formation in the striatum are independent of action potentials or Ca2+.

2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-ben↗
PubMed1994

Expression of a transgenic class II Ab gene confers susceptibility to collagen-induced arthritis.

The major histocompatibility complex (MHC) class II region is assumed to influence autoimmune diseases such as rheumatoid arthritis. In the mouse, the H-2q haplotype is associated with susceptibility to collagen-induced arthritis, while the H-2p haplotype is not. The class II A molecules of these haplotypes differ by only four amino acids in the first domain of the beta chain. To test if this difference accounts for the MHC influence on susceptibility to collagen-induced arthritis, H-2p mice were made transgenic with an Abp gene altered to resemble the Abq gene. The transgenic A beta chain hybridized with the A alpha p chain and was shown to be physiologically expressed by testing antigen-presentation capacity to Aq-restricted T cell hybridomas and with FACS analyses. These transgenic mice developed an autoimmune response to type II collagen and also collagen-induced arthritis. The data unequivocally suggest the Ab gene as a major genetic susceptibility locus for autoimmune collagen-induced arthritis.

Animals↗