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Biomedical subjects

U Hammerling

Publications and source records attributed to U Hammerling.

65 records · Page 4Linked to original sources

Endotoxin-induced serum factor controlling differentiation of bone-marrow-derived lymphocytes.

Serum from bacillus Calmette-Guerin-infected mice injected with endotoxin induces the appearance of surface immunoglobulin, Ia antigen, and complement receptor on the surface of precursor bone-marrow-derived (B) cells. While endotoxin itself causes phenotypic conversion of both thymus-derived (T) cells and B cells in vitro, the endotoxin-induced serum factor was found to be a selective inducer of B cell differentiation. Spleen cells rendered immunodeficient by removal of B cells bearing the complement receptor regained the capacity to cooperate with helper T cells and to produce antibody against red cell antigens in vitro upon upon addition of the serum factor to the culture medium. Thus, a factor that controls selective phenotypic and functional differentiation of B cells has been identified and can now be characterized,

Animals↗

Differentiation of thymocytes: evidence that induction of the surface phenotype requires transcription and translation.

Induction of thymocyte differentiation requires transcription of DNA and translation of RNA but not replication of DNA. This was determined in vitro by studying the effects of cytosine arabinoside, hydroxyurea, actinomycin D, camptothecin, cordycepin, cycloheximide, and puromycin on the induction by thymopoietin of thymocyte differentiation, measured by the acquisition of the differentiation allo-antigens TL and Thy-1 by and inducible fraction of mouse spleen cells. This requirement for transcription does not establish whether the new mRNA represents transcripts of structural genes for the thymocyte allo-antigen markers; an answer to this question awaits the results of further experiments.

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Scanning electron microscopy of tobacco mosaic virus-labeled lymphocyte surface antigens.

The study of surface antigen by immunoelectron microscopy has been hampered by the fact that thin sections of cells provide only a view of the cell perimeter in an essentially two- dimensional fashion. Although the reconstruction of the entire cell from serial sections has been accomplished (1), it remains too exacting a technique and will find only exceptional application. Carbon-platinum replicas (2) allow the inspection of larger surface areas and therefore are better suited for studying the distribution of antigens (3). But since only relatively smooth surfaces will yield stable replicas, cells with large numbers of microvilli are not amenable to this technique. Despire its limited resolution, scanning electron microscopy (SEM) seems to be the method of choice because it can provide a view of almost half of the surface of a cell close to its natural configuration, particularly after critical point or freeze drying (4, 5). Immunological-labeling methods have not yet been routinely applied to SEM although both latex spheres (6) and hemocyanin (7) have been used with some success. The optimal visual marker should possess the following properties: be of a distinctive shape, chemically stable, and have per se a low binding affinity for cell surfaces. Tobacco mosaic virus (TMV), a marker with which we are familiar in transmission electron microscopy (8), seems to meet these demands; it has rod-like shape and defined dimensions (15 x 300 nm) and in addition it can easily be distinguished from surface microvilli. As the hybrid antibody technique (9) is also applicable to TMV, we have attempted to combine such immunological labeling with SEM. We present evidence that surface antigens can indeed be visualized by SEM, using the TMV marker in conjunction with the hybrid antibody technique.

Animals↗

Isolation of a polypeptide that has lymphocyte-differentiating properties and is probably represented universally in living cells.

A polypeptide of 8500 molecular weight is described that induces the differentiation of T (thymus-derived) cell and B (bone-marrow-derived) cell immunocytes in vitro, apparently via beta-adrenergic receptors and adenylate cyclase activation. This polypeptide shows a high degree of evolutionary conservation, exhibiting close structural, functional, and immunological similarity when isolated from such diverse origins as cells of mammals and higher plants. This polypeptide was detected in animal cells, yeast, bacteria, and higher plants, and so may well be a universal constituent of living cells.

Adenylyl Cyclases↗

The ontogeny of murine B lymphocytes. I. Induction of phenotypic conversion of Ia-to Ia+ lymphocytes.

When bone marrow and spleen cells of 4 week-old mice are fractionated on a discontinuous BSA gradient, a small fraction of Ia- cells is obtained which can be induced in vitro to express the Ia alloantigen within 2 hr. This is in precise parallel to the prothymocyte induction system of Komuro and Boyse. Ia specificity is ascertained by the use of two reciprocal antisera, A.TH anti-A.TL (anti-Iak) and A.TL anti-A.TH (anti-Ias), which yield the expected reaction pattern on induced bone marrow cells of (B6 X A)F1 (Iak) and SJL/J (Ias) mice. Induction can be effected by a number of agents, such as catecholamines, prostaglandin PGE1, cAMP, bacterial endotoxin, lipid A, ubiquitin, and thymopoietin. The last requires a 100-fold higher concentration for Ia+ induction as compared to prothymocyte induction, thus implying a lower affinity for the B cell receptor than for the thymocyte receptor. Ia- to Ia+ conversion involves cells different from prothymocytes, as indicated by: 1) the specific cytolytic effect of our anti-Ia sera which were shown to be free of activity against thymocytes; 2) an additive cytolytic effect of anti-Ia and anti-Thy-1 sera; and 3) the fact that the Ia inducible cells are sensitive to pretreatment with anti-immunoglobulin and C. This finding that Ia- precursor cells are already Ig+ is of interest for the B cell ontogeny, as it implies that Ig expression precedes Ia expression.

Animals↗

Serological crossreactivity between H-Y (male) antigens of mouse and man.

Antisera to H-Y (male-specific) antigen were prepared by immunizing female mice with spleen cells from males of the same inbred strain. These antisera were used in mixed hemadsorption and cytotoxicity tests with cells of rats, guinea pigs, rabbits, and humans. The results showed that the H-Y components of all four species are antigenically related to H-Y of the mouse.

Animals↗

[Detection by hemagglutination of antibodies to group A and group E streptococci by the use of O-stearoyl derivatives of their cell wall carbohydrate-grouping antigens].

The streptococcal group A and group E cell wall polysaccharide antigens were extracted with trichloroacetic acid from the cell or cell wall and esterified with stearic acid. The stearoyl derivatives contained 5 to 8% (by weight) of the ester. Sheep or human red blood cells were sensitized with the esterified antigens and were shown to agglutinate in the presence of specific rabbit antisera. Sera from (i) children hospitalized with group A streptococcal respiratory disease and (ii) swine possessing group E streptococcal lymphadenitis were shown to possess antibody titers significantly higher than the controls. The use of the two esterified antigens as controls for each other established the specificity of the reaction in each case. The general shape of the antigen-antibody precipitin curves was not changed when the stearoyl antigens were used; however, the quantitative aspects differed markedly. Oligosaccharides which inhibit the normal antigen-antibody precipitin reaction did not inhibit the hemagglutination reaction. The adsorption of antisera with whole streptococcal cells reduced the hemagglutination titer in relation to the quantity of cells employed. Data are given on the (i) optimal concentration of stearoyl antigen for sensitization, (ii) time of adsorption of antigen to red cells, (iii) use of albumin as diluting fluid, and (iv) condition of red cells. Properties of the esterified antigens and the mechanism of the agglutination reaction are discussed. The results indicate that polysaccharide antigens of other bacteria may be esterified and employed in a similar manner.

Animals↗

Detection of a feline X-linked antigen in somatic cell hybrids. Single-cell analysis using monoclonal antibodies.

We tested the premise that monoclonal antibodies to either intracellular or membrane antigens can greatly facilitate the construction of linkage maps of mammals whose chromosomes can be introduced into rodent cells. Monoclonal antibodies against antigenic determinants of cat lymphocytes and fibroblasts were used to analyze feline antigen expression in cat-mouse somatic cell hybrid populations selected to contain the X-linked feline HPRT locus. The frequency of antigen expression as measured by fixed cell immunofluorescence (IF) assays, varied greatly within hybrid populations for all but the antigen designated as VP382. Its frequent presence in hybrid cells led to the prediction, confirmed by 8-azaguanine selection experiments, that its expression was controlled by a gene, or genes, on the feline X chromosome. The antigens identified by the rest of the antibodies segregated independently of each other in cat-mouse somatic cell hybrids and their expression appeared to be controlled by autosomal genes of the cat.

Animals↗