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Biomedical subjects

S Sartoris

Publications and source records attributed to S Sartoris.

At least 19 recordsLinked to original sources

Staphylococcal enterotoxin-dependent lysis of MHC class II negative target cells by cytolytic T lymphocytes.

The enterotoxins of Staphylococcus aureus (SE) are extremely potent activators of human and mouse T lymphocytes. In general, T cell responses to SE are MHC class II dependent (presumably reflecting the ability of SE to bind directly to MHC class II molecules) and restricted to responding cells expressing certain T cell receptor beta-chain variable (TCR V beta) domains. Recently we demonstrated that CD8+ CTL expressing appropriate TCR V beta could recognize SE presented on MHC class II-bearing target cells. We now show that MHC class II expression is not strictly required for T cell recognition of SE. Both human and mouse MHC class II negative target cells could be recognized (i.e., lysed) in a SE-dependent fashion by CD8+ mouse CTL clones and polyclonal populations, provided that the CTL expressed appropriate TCR V beta elements. SE-dependent lysis of MHC class II negative targets by CTL was inhibited by mAb directed against CD3 or LFA-1, suggesting that SE recognition was TCR and cell contact dependent. Furthermore, different SE were recognized preferentially by CTL on MHC class II+ vs MHC class II- targets. Taken together, our data raise the possibility that SE binding structures distinct from MHC class II molecules may exist.

Animals

Cell lineage-specific and developmental stage-specific controls of MHC class-II-antigen expression.

In this report we present evidence and we review data from our laboratory which indicate the genetic complexity of regulatory mechanisms controlling MHC class-II-gene expression. The MHC class-II genes can be expressed in 2 ways: in a constitutive fashion, as in B cells, and in an inducible fashion, as in macrophages, endothelial cells and certain tumors. In both cases the regulatory controls are mainly exerted at transcriptional level as a result of interactions between cis-acting regulatory DNA elements and trans-acting factors. The constitutive class-II-gene expression in B cells is under the control of developmentally regulated trans-acting factors with activator function and encoded by a series of genes, the AIR genes, one of which has been mapped in the mouse on chromosome 16. Interestingly, these regulatory mechanisms are conserved across species for at least 70 million years, because murine AIR-gene products can complement AIR gene defects of human B-cell mutants. The constitutive B-cell phenotype behaves as a dominant trait up to the plasma cell stage in which class-II-gene expression is lost because of the activation of suppressor factors which repress transcription and which, in turn, behave as a dominant trait in somatic cell hybrids between B cells and plasma cells. Thus positive and negative signals regulating class-II-gene expression may behave as dominant or recessive traits, depending upon the particular developmental stage of the cell in which they operate. The mechanisms controlling class-II expression in inducible cells are distinct from those mediating constitutive expression. Indeed, induction of these genes is not sufficient to complement AIR-gene defects in hybrids between macrophages and class-II-negative mutant B cells. In contrast, constitutive expression is dominant in hybrids between class-II-positive B cells and macrophages, suggesting that in uninduced cells class-II-gene activation does not take place more because of lack of activator factors than because of the presence of constitutive transcriptional suppressors. On the basis of these results, we propose a model for developmentally controlled MHC class-II-gene expression during ontogeny.

Animals

Inducible and constitutive MHC class II gene expression. Distinct tissue-specific genetic controls.

B cells express MHC class II Ag in a constitutive fashion, whereas macrophages can do so only after induction by a variety of exogenous stimuli. In this study we describe interspecies somatic cell hybrids between the human B cell Raji and the murine macrophage cell P388 D1. This murine cell line does not express detectable levels of class II mRNA. Phenotypic, molecular, and karyotype analysis of a series of hybrids showed that murine macrophage class II genes can be expressed in a constitutive fashion under the control of the human B cell genome. This event is the consequence of de novo accumulation of class II specific mRNA and thus probably reflects activation of transcription. In certain cases the amount of murine class II Ag expressed on the surface of the hybrid cell was significantly higher than the one observed in the parental macrophage cells after induction with IFN-gamma and was not further modified by treatment with the murine lymphokine. Reversion from a murine class II-positive to class II-negative cell surface phenotype in the hybrids correlated with reduced expression of human markers and more important with segregation of human chromosomes. Interestingly, in this case certain hybrids still expressed detectable levels of murine class II mRNA and increased levels of murine invariant chain mRNA when compared with parental P388 D1 murine macrophage cells. These results indicate that constitutive class II gene expression behaves as a dominant trait in B cell x macrophage somatic cell hybrids. Possible mechanisms responsible of the different control of class II gene expression during cell type differentiation are discussed.

Animals

Alternative splicing of HLA-DQB transcripts and secretion of HLA-DQ beta-chain proteins: allelic polymorphism in splicing and polyadenylylation sites.

HLA class II antigens are highly polymorphic cell-surface proteins involved in initiation and regulation of the immune response. Allelic sequence variation primarily affects the structure of the first external domains of alpha and beta component chains. Here we provide evidence for other types of allelic polymorphism for the genes encoding these chains. Sequences of two cDNA clones corresponding to HLA-DQB mRNAs from an HLA-homozygous cell line exhibit both alternative splicing and read-through of polyadenylylation. Furthermore, alternative splicing that deletes the transmembrane exon is associated with only a subset of HLA-DQB alleles, while the polyadenylylation-site read-through is found in a larger subset. This suggest that polymorphic cis-acting elements within the HLA-DQB gene control both processing steps. Proteins, presumably encoded by alternatively spliced mRNAs lacking transmembrane exons, are immunoprecipitated with a monomorphic monoclonal antibody directed against HLA-DQ. These proteins are found in supernatants of cultured cell lines for which secretion is predicted, but not in those of cell lines that do not contain alternatively spliced mRNAs.

Alleles

Mitochondrial DNA polymorphism in four Sardinian villages.

Polymorphism of mitochondrial DNA has been studied in two highland (Desulo, Tonara) and in two lowland (Galtellì, Orosei) Sardinian isolates, formerly subjected to different selective pressure due to malaria, and in 103 individuals from Northern Italy (Bergamo area), where malaria never appeared to be endemic. Two mitochondrial restriction endonuclease patterns (morphs) never described before have been found, one in the Bergamo and Orosei samples, and the other one only in Orosei. Four new mitochondrial types (mitotypes) due to different combinations of morphs have been identified; two of them have been found only in Sardinia, but with such a low frequency that they cannot be defined as typical Sardinian mitotypes. One mitotype (BamHI-morph 3, MspI-morph 4, AvaII-morph 9 and HaeII-morph 1) showed a significantly higher frequency in the highland rather than in the lowland Sardinian villages or in the Bergamo area. Since this mitotype has been found at a relatively high frequency in Central and Southern Italy, while it has been reported to be rare in Caucasians of Central European origin and absent in other ethnic groups (Africans, Chinese, Japanese and Israeli Jews), we suggest it may represent an ancient Mediterranean type. The analysis of these data suggests that drift or other evolutive forces different from malaria might be the major cause of mitochondrial DNA variation in Sardinia.

DNA, Mitochondrial

A rapid and improved method for generating cDNA libraries in plasmid and phage lambda vectors.

We have developed a fast and efficient procedure for generating cDNA libraries in plasmid or phage lambda vectors. We used Mo-MuLV reverse transcriptase to synthesize the first strand and directly added Escherichia coli DNA polymerase I with RNase H to synthesize the second strand. A special advantage of our procedure is the use of oligodeoxynucleotide adapters to insert the cDNA into the vector, avoiding the use of methylating enzymes and subsequent digestion with massive amounts of restriction endonucleases. This also obviates the need to tail the cDNA molecules with homopolymers, simplifying subsequent procedures such as sequencing or transfer to other vectors. Finally, we have used a rapid screening procedure to isolate full-length clones with oligodeoxynucleotide probes recognizing conserved regions at the 5' termini of the mRNA. The system is ideal for cloning and analyzing polymorphic alleles of genes, such as those of the major histocompatibility complex.

Bacteriophage lambda

[Actinomycetales in cutaneous pathology].

After discussing the classification of the Actinomycetales, the authors give a detailed description of the epidemiology and particularly of the clinical features of cutaneous diseases induced by genera such as Actinomyces, Norcardia and Streptomyces. Histopathological and immunological studies, together with laboratory and differential diagnostics, are also examined. In the last section, present therapeutic treatments are briefly outlined.

Actinomyces

[Eccrine-nevus (author's transl)].

The eccrine nevus is an exceptional entity, formed by well-differentiated adnexa which can be classified as hamartoma or organoid nevus. It is characterized by a high degree of structural maturation. The authors report a case of eccrine nevus observed in a 32-year-old woman and compare clinical and histological features of the lesion with literature data.

Adult