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Biomedical subjects

G Gaudernack

Publications and source records attributed to G Gaudernack.

144 records · Page 8Linked to original sources
PubMed1982

In vitro differentiation of human monocytes. Differences in monocyte phenotypes induced by cultivation on glass or on collagen.

We demonstrated that the in vitro differentiation of human peripheral blood monocytes to macrophages is dependent on the environment and conditions of monocyte culture. Cultivation of monocytes on glass or microexudate-coated glass gave rise to cells resembling foreign body granuloma macrophages. After an initial rise in Fc receptor- and C3 receptor-mediated phagocytosis, a progressive loss of Fc receptor expression and C3-mediated ingestion were observed. The monocyte surface antigens recognized by the anti-human monocyte monoclonal antibodies 1D5 and 63D3 were lost from the surface of the majority of cells cultured on glass and microexudates. A subpopulation of Fc receptor-positive cells that were 1D5 and 63D3 positive was retained in fully differentiated cell populations. In comparison, monocytes cultivated on collagen matrices gave rise to highly phagocytic cells resembling human resident tissue macrophages. Both Fc- and C3-mediated phagocytosis were enhanced and remained so during the entire length of culture. The surface antigens recognized by the 1D5 antibody, expressed on all freshly seeded monocytes, was maintained on the macrophages. The antigen recognized by the 63D3 antibody was not expressed on mature cells. The present evidence would indicate that variations in expression of phagocytic receptors and the surface antigens 1D5 and 63D3 can be ascribed to the stage of development of the macrophage or its stage of activation, rather than to independent subsets of mononuclear phagocytes.

Antibodies, Monoclonal↗
PubMed1982

Antibodies to and elevations of beta 2 microglobulin in the serum of ankylosing spondylitis patients.

Antibodies to beta 2 microglobulin are found in systemic lupus erythematosus patients and are important in the lymphocytotoxic reactions of sera from such patients. In this study, beta 2 microglobulin antibodies were measured with the use of an enzyme-linked immunosorbent assay with purified beta 2 microglobulin antigen and peroxidase-labeled anti-human IgG or IgM. IgG antibodies to beta 2 microglobulin were found in 68% of 22 patients with ankylosing spondylitis. This incidence was higher than the 5% in 80 controls (P less than 0.01) and similar to the 71% incidence found in 35 patients with systemic lupus erythematosus. Eleven (27%) of 41 patients with rheumatoid arthritis had elevated levels of antibodies to beta 2 microglobulin (P less than 0.01). The mean antibody levels expressed in enzyme units were 0.125 for patients with ankylosing spondylitis, 0.157 for those with systemic lupus erythematosus, 0.101 for those with rheumatoid arthritis, and 0.067 for controls. IgM anti-beta 2 microglobulin was not significantly different from controls. A competitive binding assay with enzyme-labeled beta 2 microglobulin was used to determine serum beta 2 microglobulin. These values were also found to be elevated in 48% of patients in all 3 disease categories (P less than 0.01). Beta 2 microglobulin antibodies and serum beta 2 microglobulin did not correlate with each other, renal diseases or antinuclear antibodies in patients with systemic lupus erythematosus, with rheumatoid factor or severity of articular disease in patients with rheumatoid arthritis, or with peripheral arthritis or iritis in those with ankylosing spondylitis. Although antibodies to beta 2 microglobulin might reflect a general disturbance of immune regulation in patients with systemic lupus erythematosus, their presence in those with ankylosing spondylitis, a disease closely associated with a specific HLA allotype and not usually associated with formation of autoantibody, suggests that they might play a role in the pathogenesis of the latter disease.

Adult↗
PubMed1982

Monoclonal hybridoma antibodies to human amyloid related protein SAA.

Problems concerning isolation and characterization of the amyloid related serum protein SAA in a pure form prompted us to make monoclonal antibodies to the protein. Protein SAA isolated by gel filtration under dissociating conditions was used for immunization of BALB/c mice, and spleen cells from a mouse producing high titred antiserum to SAA were fused with cells from the mouse plasmacytoma line P3U1. Antibody specificity to various preparations of protein SAA was tested using an indirect enzyme-linked immunosorbent assay. Monoclonal antibodies with specificity for SAA were obtained in addition to antibodies which reacted with both SAA and the related amyloid protein AA. Antibodies specific for one of the apoC proteins of the lipoprotein fraction were also produced showing that the SAA preparation used for immunization was contaminated with apoC proteins.

Amyloid↗
PubMed1981

Rapid identification of human B-lymphocytes and monocytes with rhodamine-labeled Brucella melitensis.

The bacterium Brucella melitensis has been shown to bind selectively to human B-lymphocytes. The specificity of this binding can be exploited in a rapid technique for the determination of human B-lymphocytes and monocytes by use of rhodamine-labeled Brucella melitensis. This fluorescent labeled regent is simple to use and provides highly specific identification of human B-lymphocytes and monocytes, as demonstrated in a series of experiments characterizing known cellular markers on T- and B-lymphocytes, their separated cellular subpopulations and lymphoblastoid cell lines. The use of fluorescence conjugated bacteria greatly simplifies the application of this highly specific technique and makes its use more practical in routine screening for B-cell and monocyte populations.

Antigens↗
PubMed1980

Immunization with the light chain and the VL domain of the isologous myeloma protein 315 inhibits growth of mouse plasmacytoma MOPC315.

Prior immunization of BALB/c mice with free light chains from myeloma protein 315 (L315) and its variable domain (VL315) inhibited the growth of subcutaneously injected MOPC315 tumour cells. The growth suppression observed after immunization with L315 was equivalent to that which resulted from immunization with the complete M315. VL315 and non-polymerized L315 did not elicit specific antibodies. Polymerized L315 induced both suppression of MOPC315 growth and antibodies specific for free L315; however, these antibodies did not react with the complete M315, nor were they absorbed by MOPC315 tumour cells. The data indicate that the suppression of tumour growth was mediated by specifically sensitized cells acting in the absence of antibodies against M315 or L315. Immunization with the variable domain of the heavy chain from M315 (VH315) had no effect on the growth of MOPC315. The M315 fragments and subunits that induced growth suppression were thus identical with those capable of inducing T helper cells in BALB/c mice.

Animals↗
PubMed1980

T- and B-cell-independent activation of syngeneic macrophages by murine sarcoma cells.

Mouse peritoneal macrophages were achieved by cocultivation with syngeneic sarcoma cells. The tumor cells died progressively during the cocultivation, leaving highly activated marcophages. Because of great changes in macrophage morphology during the activation, special efforts were made to identify the activated cells as macrophages by their ability to phagocytose latex and to bind opsonized sheep red cells to C3 and Fc receptors and by indirect immunofluorescence with an antimacrophage antiserum. Activation was evaluated by morphology and incorporation of [14C]glucosamine. The activation was found to be independent of the presence of T-cells, B-cells, and immunoglobulin bound to tumor cell surfaces. This was shown by removal of T-cells from the system by treatment with anti-theta and complement and by use of nude mice as the macrophage source and for tumor maintenance. Similarly, B-cells were removed by treatment with anti-immunoglobulin and complement as well as adherence to anti-immunoglobulin-coated plastic dishes. Immunoglobulin bound to tumor cells was removed by trypsinization and by elution at low pH. Culture supernatants from tumor cells and cell-free tumor ascites fluid also induced some activation of the macrophages. This activation differed from the coculture activation in both the extent and kinetics of morphological changes and gave only a small increase in [14C]glucosamine incorporation.

Animals↗
PubMed1979

The plasma cell differentiation antigen PC.1 is absent in CH3/Tif and present in C3H/HeJ.

(C3H/TifxDBA/2)F1 mice, immunized with viable BALB/c plasmacytoma MOPC315 cells, produce antibodies directed against a cell-surface antigen. The strain and tissue distribution of this antigen was identical to that of the plasma cell differentiation alloantigen PC.1. The antigen is absent in the mouse strain C3H/Tif but is present in the closely related substrain C3H/HeJ. This is the third difference between surface structures of the B-cell lineage of C3H/Tif and C3H/HeJ mice.

Animals↗
PubMed1977

Ligand-induced redistribution and augmentation of surface-bound myeloma protein on MOPC 315 plasmacytoma cells.

The dinitrophenyl (DNP)- and trinitrophenyl (TNP)-binding IgA(lambda2) myeloma protein M315, bound on the surface of MOPC315 mouse plasmacytoma cells, was redistributed into spots, patches, and, more rarely, into caps by TNP14-BSA and by divalent but not monovalent anti-M315 antibodies. Antiserum to the L-chain of M315 (L315) induced similar redistribution of L315 bound on the surface of variant cells that only produced L315. The spots were much larger and more brilliant when the cells were incubated with the ligands at 37 degrees C than at 4 degrees C. Redistribution of M315 also occurred on M315-producing cells in peritoneal diffusion chambers incubated in BALB/c mice producing antibodies against the M315 idiotype. The clearance of immune aggregates and the regeneration of new surface-bound M315 in diffusion chambers were much slower for MOPC315 cells than that reported for B lymphocytes. The total pool of M315 was 1.9 pg per cell (about 8 x 10(6) 7S molecules), but only an average of 6 x 10(3) [125I]TNP-BSA molecules were bound on the surface of each MOPC cell at 4 degrees C. The amount of surface-bound TNP-BSA increased eightfold when the cells were preincubated at 37 degrees C with rabbit anti-mouse IgA; at 4 degrees C the increase was only twofold. The data indicate that multivalent ligands specific for M315 induce an accumulation of M315 on the cell surface that correlates with secretion; the immediate precursors of secreted myeloma protein may be arrested in their transit through the membrane by the ligands.

Animals↗
PubMed1977

Production of BALB/c anti-idiotypic antibodies against the BALB/c myeloma protein 315 does not require an intact ligand-binding site.

To determine whether the ligand-binding site of the BALB/c myeloma protein 315 is essential for the anti-idiotypic response in syngeneic animals, 17 BALB/c mice were immunized with M315 that had been affinity-labeled with bromo-acetyl-DNP-L-lysine (BADL). Essentially all the active sites of M315 were blocked by the affinity label. Fourteen mice produced antibodies that reacted with an idiotypic determinant localized in the Fv fragment of M315, but this idiotype was not part of the DNP-lysine-binding site, and it was absent from L315 and H315 chains. Two groups of BALB/c mice were immunized with nonaffinity-labeled M315, to determine whether the same idiotype was recognized with this immunogen. All animals in the group that received the most prolonged immunization produced antibodies that could be divided in two populations: about 75% were directed against the site-associated idiotype, and the rest reacted with the nonsite idiotype. The other group produced antibodies exclusively specific for the site. Thus, the site-associated idiotype of M315 is not essential for the antibody response of BALB/c mice against M315, and M315 carries at least two different idiotypes that can be recognized by B cells of syngeneic animals.

Animals↗
PubMed1975

Studies on PIVKA-X.

The incorporation of 14C-glucosamine and 3H-leucine into PIVKA-X and factor X has been studied in Macaca mulatta by means of a monospecific precipitating antiserum. No difference in the incorporation into PIVKA-X and factor X was found. The half-lives of PIVKA-X and factor X were similar (about 30-35 h and 27-31 h respectively) indicating that the vitamin K-induced modifications did not alter the survival of protein. A Russell's-viper venom/cephalin test system for factor X was more sensitive to inhibition by PIVKA-X than was a tissue thromboplastin-based system.

Adsorption↗
PubMedDate not supplied

Mutation analysis of TP53 exons 5-8 by automated constant denaturant capillary electrophoresis.

DNA fragments melt characteristically according to their nucleotide sequence and length, when exposed to denaturants such as temperature, urea or formamide. Small differences within a defined sequence, like a base mutation, will result in a slightly different melting behavior of the aberrant DNA fragment compared to that of the wild type sequence. This feature has previously been exploited for mutation detection by constant denaturant capillary electrophoresis (CDCE). In this report, we describe an automated approach (ACDCE) using a commercially available apparatus (ABI 310 Genetic Analyzer) to analyze mutations in exons 5-8 of TP53. The running conditions were determined by temperature titration of the fragments on the apparatus, and an operating sensitivity showed that 0.1% mutated alleles could be detected against a background of wild-type alleles. Up to 48 samples can be analyzed by ACDCE without any need for operator intervention. The apparatus is commercially available, and there is no need for instrument modification. To our knowledge this is the first report on the analysis of TP53 exons 5-8 by ACDCE.

Automation↗
PubMedDate not supplied

Development of a clinical grade procedure for generation of mRNA transfected dendritic cells from purified frozen CD34(+) blood progenitor cells.

Enriched CD34(+) peripheral blood progenitor cells (PBPC) are frequently used as stem cell support in cancer patients following high dose therapy. Since precursor dendritic cells (DCs) originate from haematopoietic progenitor cells, purified CD34(+) cells might also serve as starting cells for ex- vivo production of DC. In the present study we developed a clinical grade procedure for ex- vivo production of DC derived from enriched CD34(+) cells. Different concentrations of CD34(+) cells were grown in gas-permeable Teflon bags with different serum-free and serum-containing media supplemented with GM-CSF, IL-4, TNF-alpha, SCF, Flt-3L and INF-alpha. Serum-free CellGroSCGM medium for 7 days followed by CellGroDC medium in 7 days gave the same results as serum-containing medium. After incubation the cultured cells containing immature DCs were concentrated and transfected with tumour mRNA from human prostate cancer cell lines employing a highly efficient electroporation procedure. Thawed transfected DCs were able to elicit primary T-cell responses in vitro against antigens encoded by the prostate cancer mRNA as shown by ELISPOT assay using mock-transfected DCs as control. Our results show that frozen enriched CD34(+) cells can be an alternative and efficient source for production of DCs for therapeutic purpose.

Antigens, CD34↗
PubMedDate not supplied

Early stage carcinoma of the uterine cervix. Effects of intracavitary radium treatment on lymphoid cells in blood and pelvic lymph nodes.

Sixteen patients with early stage carcinoma of the uterine cervix treated with primary radical hysterectomy and pelvic lymphadenectomy were compared with 17 patients who four to six weeks before the operation received intracavitary treatment with radium. The calculated radiation dose to the pelvic wall was approximately 10 Gy. The distribution of lymphoid cells in blood and pelvic lymph nodes was studied by an indirect immunofluorescence technique using monoclonal antibodies. The radium treated group showed a significant reduction of circulating OKT4+ (T helper) and OKT8+ (T suppressor/cytotoxic) lymphocytes. The number of Leu7+ (natural killer) cells and 1D5+ cells (monocytes) was not changed, but the ratio between monocytes and T cells was increased after radium therapy. In cell suspensions obtained from the pelvic lymph nodes, the radium treatment induced a significant reduction of the OKT4+ cell fraction. It is concluded that this low dose rate regimen of intracavitary treatment induces changes in the immune system which are of the same type as those seen after external field irradiation.

Adult↗