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T Hansen

Publications and source records attributed to T Hansen.

At least 217 records · Page 12Linked to original sources
PubMed1990

Regulation of insulin receptor functions by a peptide derived from a major histocompatibility complex class I antigen.

A 25 residue peptide, Dk-(61-85), derived from the alpha 1 domain of a murine MHC class I molecule (H-2Dk), enhances cellular glucose uptake, prolongs the effect of insulin, and inhibits insulin receptor internalization without affecting insulin binding or dissociation. Full effect of the peptide is obtained at 10-100 microM. The magnitude of the peptide-mediated enhancement of glucose uptake is insulin dependent and is at maximum approximately 50% above that of full insulin stimulation, excluding a merely insulinomimetic action of the peptide. Dk-(61-85) does not interact directly with the glucose transporter molecule. Furthermore, the peptide-mediated inhibition of insulin receptor internalization results in 2-3 times more receptors in the plasma membrane. The peptide also causes hypoglycemia in rats. The biological activity of Dk-(61-85) suggests that an important nonimmunological role of MHC class I molecules is to affect some of the key functions of ligand-activated receptors.

Amino Acid Sequence↗
PubMed1989

Nicardipine after spinal cord compression in the lamb.

To study the effects of the calcium channel blocker nicardipine on spinal cord blood flow and spinal evoked potentials, the following study was undertaken. After cord compression, which was productive of paraparesis, nicardipine was administered intravenously in 10 anesthetized lambs. Ten control animals were subjected to compression but received saline instead. Nicardipine produced a significant decrease in mean arterial pressure when compared to the control group. Thirty minutes after compression, spinal cord blood flow also was lower in the nicardipine group compared with controls. Spinal evoked potentials did not recover after compression in either group.

Animals↗
PubMed1989

Inhibition of insulin receptor phosphorylation by peptides derived from major histocompatibility complex class I antigens.

Peptides from the alpha 1 region (residues 61-85) of the D and K molecules of the major histocompatibility complex class I antigens inhibit insulin-induced tyrosine kinase activity of the purified human insulin receptors (IRs) as measured both by autophosphorylation and IR-mediated substrate [poly(Glu,Tyr)] phosphorylation. Half-maximal effect of the Dk-(61-85) peptide on IR autophosphorylation is obtained at 1.2 microM, and almost complete inhibition of IR kinase activity is obtained at 10 microM peptide. The corresponding K kappa-(61-85) peptide has a significantly weaker effect on autophosphorylation. No such effects are observed with nine peptides of similar length, but unrelated to major histocompatibility complex class I antigens. Neither of the major histocompatibility complex class I-derived peptides has any effect on the constitutively active kinase of a genetically engineered cytoplasmic IR domain. Further, insulin binding to IR is unaltered in the presence of the major histocompatibility complex class I-derived peptides. The inhibitory activity of the peptides on insulin-induced IR phosphorylation facilitated the observation that IRs require insulin to become substrate for an independent tyrosine kinase. In the presence of an inhibitory peptide, the constitutively active cytoplasmic IR kinase domain only phosphorylates the intact IR in the presence of insulin. We conclude that the tyrosine kinase activity of IRs may be altered by peptide interaction at an allosteric site and, moreover, IRs require insulin to assume a conformation permitting phosphorylation by an independent kinase.

Allosteric Site↗
PubMed1989

Methylprednisolone in spinal cord compression.

In acute nonsurvival studies, eight anesthetized lambs were subjected to cord compression at T13 by means of an epidural balloon distended to a pressure of 200 mm Hg for 40 minutes. Subsequent to withdrawal of the balloon, each animal received 30 mg/kg of methylprednisolone succinate in an intravenous bolus followed by a continuous infusion of 10 mg/kg/hr for the duration of the experiment. Spinal cord blood flow (SCBF) and spinal evoked potential (SEP) determinations were obtained sequentially prior to, during, and at 1/2, 1 1/2, and 2 1/2 hours following compression. In spite of the absence of ischemia following compression, SEPs failed to recover. Methylprednisolone had no apparent effect on blood flow or on the recovery of SEPs when compared with results in ten control animals that received saline alone.

Animals↗
PubMed1989

Two cytotoxic human-human hybridoma antibodies to HLA: TrAH10 (anti A3.1) and TrAG2 (anti B7, Bw42).

Two cytotoxic human-human hybridoma IgM antibodies to HLA were generated by EBV transformation of PBMC from multiparous women and fusion of EBV transformed cells with the human fusion partners KR4 or KR12. Both mAbs required the sensitive immunomagnetic cytotoxicity method to display killing of freshly prepared PBMC. One mAb (TrAH10) was specific for HLA-A3. Strikingly, TrAH10 reacted much more strongly with lymphoblastoid cell lines of HLA-A3.1 than of the rare variant HLA-A3.2, previously detected by cytotoxic T cells. Thus, in the microcytotoxicity test, the titer of concentrated TrAH10 was approximately 2000 times higher for A3.1 as compared to A3.2, and a clear difference was also observed in radioimmunoassay. Since the two HLA-A3 variants differ by only two amino acids at positions 152 and 156 of the alpha 2-domain's alpha-helix, the epitopes defined by the mAb TrAH10 and HLA-A3.1 specific cytotoxic T cells must be closely related. The observations with TrAH10 suggest that the HLA polymorphism detected by human mAbs may turn out to be as extensive as the T-cell defined HLA polymorphism. The other mAb (TrAG2) bound B7 and Bw42 with equal strength, and in addition bound weakly to some cells that were Bw22 or B39. Magnetic polymerbeads coated with affinity purified human mAbs TrAH10 or TrAG2 formed rosettes with EBV transformed cells carrying relevant HLA antigens; however, rosette formation with freshly isolated PBMC was very weak and unsuitable as a typing assay.

Antibodies, Monoclonal↗
PubMed1989

Direct HLA typing by rosetting with immunomagnetic beads coated with specific antibodies.

Paramagnetic monodisperse polymer beads were coated with purified human hybridoma antibodies specific for polymorphic HLA determinants. The mAb-coated beads bound specifically to cells from individuals with the relevant HLA type. The rosettes formed by beads and cells were isolated with a magnet. Rosette formation was evaluated microscopically and used as criterion for positive typing. By this rosette assay, fast and reliable HLA typing of whole blood was possible.

Antibodies, Monoclonal↗
PubMed1989

Annular erythema in identical twins.

A case of superficial gyrate erythema that was indistinguishable from erythema annulare centrifugum occurred in identical twins. We propose to include a familial cause among the known causes of erythema annulare centrifugum rather than considering this a separate entity, "familial annular erythema."

Biopsy↗
PubMed1988

Influence of major histocompatibility complex H-2LD class I molecules on spleen colony-forming units.

To test whether the major histocompatibility complex class I genes are involved in the regulation of hemopoiesis, the stem cell activities of BALB/c-H-2dm2 (Dm2) mice, which are defective in the expression of H-2L antigens, have been compared with those of the wild-type, BALB/c-Kh, in in vivo and in vitro stem cell assays. In spleen colony-forming unit assays, Dm2 as hosts consistently supported a smaller number of colonies than did BALB/c-Kh. However, both Dm2 and BALB/c-Kh supported a comparable number of colonies in in vitro granulocyte-macrophage colony-forming unit and erythroid colony-forming unit assays. These observations together suggest that the mutation in Dm2 has not affected the hemopoietic potential of the stem cells but may probably affect the hemopoietic microenvironment for the development of the stem cells.

Animals↗
PubMed1988

Four cytotoxic human-human hybridoma antibodies that react with HLA-B27.

PBMC were isolated from a multiparous woman with HLA-B27 specific Abs in her serum. The HLA type of the donor was A2,9:B7. The PBMC were EBV transformed, and four cell lines making cytotoxic Abs to HLA-B27+ cells prepared. Hybridomas were constructed by fusing the EBV lines with the human fusion partner KR4. All four mAbs were of IgM isotype. One mAb (TrBH12) reacted specifically with B27+, B37+ and Bw47+ lymphoblastoid cell lines and with all B27+ PBMC except for a rare variant so far found only in one Norwegian family. Another mAb (Tr3B6) was cytotoxic for all B27+ cells tested, including the TrBH12- variant; in addition, it showed weaker cross-reactions to Bw42, B49 and a cell line with the probable phenotype B7,38. Supernatant from the Tr3B6 hybridoma was tested in lymphocytotoxicity against a panel of 658 individuals, 141 of whom were B27+. With this panel, Tr3B6 showed perfect correlation with HLA-B27. The two last mAbs (TrCG10 and TrBF1) reacted with all B27+ cells tested, but in addition showed quite extensive cross-reactions.

Antibodies, Monoclonal↗
PubMed1988

A cytotoxic human-human hybridoma antibody (TrH6) specific for HLA-DRw52.

TrH6 is a new human-human hybridoma antibody (Ab) of IgM, lambda isotype. At concentrations of 17-5000 ng IgM/ml, TrH6 killed twelve out of fourteen lymphoblastoid cell lines that expressed DRw52, but none of the nine DRw53 homozygous lines. Both DRw52+ cells (LUY, TAB) not killed by TrH6 were DR8+, suggesting that these lines express a structural variant of DRw52 linked to DR8. The murine cytotoxic DRw52-specific mAb 7.3.19.1 (Koning et al. 1984) exhibited the same pattern of reactivity with DRw52+ cells as TrH6. However, unlike TrH6, 7.3.19.1 cross-reacted weakly with two DR7+DRw53+ homozygous cell lines, suggesting that the TrH6 epitope has a more limited distribution. The reactions with a panel of cells with known deletions in the HLA-region supported the conclusion that TrH6 is specific for DRw52. Further evidence was obtained in a competition RIA, where the murine mAbs L243 (anti-DR monomorphic) and 7.3.19.1 (anti-DRw52) inhibited binding of radiolabeled TrH6 to DRw52 homozygous cells. TrH6-coated magnetic beads provided a simple rosette-assay for typing of "buffy-coat" leukocytes for the TrH6 epitope.

Antibodies, Monoclonal↗
PubMed1987

A human-human hybridoma (Tr7E2) producing cytotoxic antibody to HLA-DQw1.

A human-human IgM (lambda) hybridoma antibody (called Tr7E2) was constructed by fusing Epstein-Barr virus-transformed cells from a multiparous woman with the human fusion partner KR12. By eosin exclusion microcytotoxicity the monoclonal antibody killed 12 of 13 human leukocyte antigen DQw1-bearing lymphoblastoid cells. No reaction was seen with any of 19 DQw1-negative cells. The single DQw1+ cell line that was not killed by Tr7E2 was the homozygous cell called 9WS 806 TAB (DR8,8; DQw1,1) of Japanese origin. The radioimmunoassay indicated that this result was probably not because of a decreased expression by this cell of DQ antigens, and these cells were killed by the mouse monoclonal antibody Genox3.53G2a5, reported to be specific for DQw1. Thus, the Tr7E2- cell line TAB probably expresses a novel structural DQw1 variant. Of 213 Norwegians, 107 were Tr7E2+ Genox+; none expressed only one of these epitopes. The putative split is, therefore, probably very rare in this population. Monodisperse magnetic polymer beads coated with Tr7E2 formed rosettes selectively with peripheral blood mononuclear cells from DQw1-positive individuals, suggesting a new approach to typing for class II antigens.

Antibodies, Monoclonal↗
PubMed1987

A human-human hybridoma antibody (TrB12) defining subgroups of HLA-DQw1 and -DQw3.

We have constructed an IgG, kappa human--human hybridoma Ab(TrB12), which precipitates a molecule consisting of two polypeptides of about 33 and 27 kD in size. TrB12 reacted with; (1) 7 out of 10 DQw1-positive cell lines in IIF, and all 10 in a rosette-assay; (2) 5 out of 12 DQw3-positive cells, both in IIF and the rosette-assay; (3) none of 4 DQw2 homozygous cells. Detergent cell lysate of DQw1 homozygous cell lines contained antigens that cross-linked the mouse monoclonal antibody Genox 353 G2a-5 (anti-DQw1) and TrB12. TrB12 competed with the mouse DQw3-specific monoclonal antibody IVD-12 for binding to DQw3 homozygous cells. The data imply that the TrB12 epitope is associated with molecules that carry DQw1 and DQw3 serological specificities. By radioimmunoassay, TrB12 and the mouse monoclonal antibody IIB3 divided both DQw1- and DQw3- bearing cell lines into three phenotypic groups: (1) TrB12+IIB3hi, (2) TrB12-IIB3lo, and (3) TrB12-IIB3-. For DQw1 the results suggest that the first two groups represent structural variants but the third group may reflect low expression of DQw1. For DQw3 the evidence suggests that all three phenotypes represent structural variants. DQw3 has previously been divided into two serologically defined alleles, TA10+IIB3- and TA10-IIB3+. The TrB12+IIB3hi and TrB12-IIB3lo variants of DQw3 described in this study probably represent novel subgroups of the TA10-IIB3+ allele.

Alleles↗
PubMed1987

Two M315 idiotopes defined by isologous monoclonal antibodies: one depends on germline and the other on mutated murine lambda 2 light chain sequences.

We have previously reported that T helper cells of BALB/c mice recognize the unique mutated sequence Phe94, Arg95, Asn96 of the lambda 2 L chain of isologous (BALB/c) myeloma protein 315. Here we study two Id (Id-315.1.4 and Id-315.TH) of the DNP-Lys binding M315 defined by two monoclonal isologous anti-Id Ab (Ab 2-1.4 and Ab 2-TH). Both Id were (1) totally expressed by Fv-315, but not by free unpaired V domains, (2) specifically dependent on VH-315, since lambda 2-315 recombined with four other H chains did not express the Id, (3) related to the hapten-binding site because their expression was blocked by the haptens DNP-Lys and DNP-Gly, and (4) topographically related because Ab 2-1.4 and Ab 2-TH competed with each other for binding to M315. The contribution of lambda chain V regions was studied with the aid of reconstituted Ig molecules of H-315 paired with lambda 1, lambda 2, and lambda 3 L chains. Id-315.TH was expressed equally well by reconstituted Ig containing three different lambda 2 chains (lambda 2-5-7, lambda 2-T952, and lambda 2-315), but its expression was profoundly reduced when H-315 was associated with lambda 3-SAPC15 or lambda 1-J558 L chains; it therefore depended upon amino acids encoded by germline lambda 2 genes. By contrast, Id-315.1.4 was only restored by the lambda 2-315 chain paired with H-315. Since lambda 2-5-7 and lambda 2-T952 differ from lambda 2-315 at positions 38, 94, 95, 96, and 98 or 99, respectively, Id-315.1.4 probably requires the unique mutated amino acids Phe94, Arg95, Asn96 of lambda 2-315. This resembles the effects on Id expression of previously reported unique amino acids of the D region. We failed to confirm that hyperimmunization of BALB/c mice with Ab 2-1.4 cross-linked to KLH induced M315-like Ab. The results are discussed in terms of the contribution of the third hypervariable loop of lambda chains to Id and the immunogenicity of isologous Ig.

Amino Acid Sequence↗
PubMed1987

A human-human hybridoma producing cytotoxic antibody to HLA-B15, cross-reacting with B17, B5, B35 and B18.

Mononuclear blood cells from a multiparous woman were transformed with Epstein Barr virus, and a cell line (Tr2D8) producing anti-HLA antibody was obtained. This cell line was immortalized by hybridization to the human fusion partners KR4 and KR12. While the EBV line died after 7 months, the hybridomas have remained stable for 13 months. The EBV line supernatant (40 micrograms IgM/ml) lysed peripheral blood mononuclear cells (PBMC) bearing B15, B17, B5 and B35. Consistent lysis of B18 bearing cells was only observed with lymphoblastoid cell lines. The supernatant from the Tr2D8 (EBV line X KR4) hybridoma (2.7 micrograms IgM/ml) only lysed B15 bearing PBMC. At a concentration of 13.5 micrograms IgM/ml, the hybridoma antibody lysed lymphoblastoid cell lines bearing B15, B17, B5, B35 and B18.

Antibodies, Monoclonal↗
PubMed1987

Simple rosette assay for HLA-B27 typing of whole blood samples.

Two human IgM monoclonal antibodies (TrBH12 and Tr3B6) specific for HLA-B27 were coupled to magnetic monodisperse polymer beads by spontaneous adsorption. A few minutes after mixing the beads with 50 microliter of whole blood, rosettes had formed specifically with leucocytes from HLA-B27 positive individuals. In this simple way, reliable typing for HLA-B27 could be conducted rapidly (in about 20 min) by evaluation of rosettes with an inverted microscope and ordinary light. The method requires purified monoclonal antibodies.

Antibodies, Monoclonal↗