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F Rupp

Publications and source records attributed to F Rupp.

30 records · Page 2Linked to original sources
PubMed1991

Structure and expression of a rat agrin.

Agrin is a component of the basal lamina that causes the aggregation of acetylcholine receptors on cultured muscle fibers. An agrin cDNA clone isolated from electromotor neurons of a marine ray was used to characterize the corresponding cDNAs from a rat embryonic spinal cord library. Analysis of a set of clones predicts a 1940 amino acid protein containing 141 cysteine residues. The predicted protein has nine domains homologous to protease inhibitors, a region similar to domain III of laminin, and four epidermal growth factor repeats. The agrin gene is expressed in rat embryonic nervous system and muscle. The rat agrin protein is concentrated at synapses, where it may play a role in development and regeneration.

Agrin↗
PubMed1989

Cloning, analysis, and expression of murine perforin 1 cDNA, a component of cytolytic T-cell granules with homology to complement component C9.

The nucleotide sequence coding for the cytotoxic T-lymphocyte (CTL) protein perforin 1 (P1) has been determined and the corresponding protein sequence has been derived. Murine CTL cDNA libraries contained in the vector lambda gt11 were screened by using a monospecific antiserum to purified P1. Three recombinant phages were isolated and their cDNA inserts were sequenced. The derived protein sequence contains 554 amino acids and displays, as expected, considerable homology with certain functional domains in the complement components C9, C8 alpha, C8 beta, and C7. The identity of P1 cDNA clones was verified by prokaryotic expression and the reactivities of antisera produced to the expressed proteins. In addition, antisera were produced to two synthetic peptides located in the center and C-terminal portions of P1. All antisera reacted with purified P1. In Northern blot analyses, P1 cDNA probes recognized a 2.9-kilobase mRNA only in CTL. Perforin mRNA was found in all cloned CTL and in all mixed lymphocyte reactions that gave rise to cytotoxic cells. Perforin mRNA was also detected in virus-specific CTL that had been generated in vivo and isolated from liver tissue of mice infected with lymphocytic choriomeningitis virus. The cell-specific expression of perforin is consistent with its postulated role in cytolysis.

Amino Acid Sequence↗
PubMed1988

Expression of T cell receptor gamma-chain in murine cytotoxic T cells: analysis of a highly transcribed nonrearranged gene cluster.

The murine T cell receptor gamma-gene family is organized in 4 gene clusters which encode variable, joining and constant gene segments. Because the function of the gamma-gene products is still unclear, there is great interest in functional rearrangements of these genes. We searched for new variable gamma-gene segments in the gamma 4-gene cluster. The mapping and sequencing of a supposed new variable segment of a functionally spliced transcript from a cDNA library of the cytotoxic T cell clone 3F9 was attempted. The results show that the supposed variable segment is a transcribed noncoding germline sequence 5' adjacent to the gamma 4-joining segment. With the gamma 4-constant sequence as a probe, strong signals could be shown in Northern analysis of the original T cell clone 3F9 and in other virus-specific T cell clones. Since all the T cell clones investigated did not show any rearrangement in the gamma 4-gene cluster, the observed high transcriptional activity in this region could be involved functionally or as an intermediate in T cell receptor rearrangement.

Animals↗
PubMed1987

T cell receptor variable gene usage in a specific cytotoxic T cell response. Primary structure of the antigen-MHC receptor of four hapten-specific cytotoxic T cell clones.

The primary structure of the alpha and beta chains of the T cell antigen receptor in four cytotoxic T cell clones specific for N-iodoacetyl-sulfonic-naphthyl-ethylene-diamine (AED)-haptenated target cells displaying a particular class I MHC molecule has been determined. Two of the T cell clones, 8/10-2 and 5/10-20K, recognize AED-modified targets in association with H-2Kb, while the other two clones 5/10-20D and C9 react with AED-modified cells in the context of H-2Db. Comparison of the nucleotide sequences of both the alpha and beta chain cDNAs and their deduced protein sequences indicates that a specific variable gene segment was not used to recognize the hapten and/or class I gene products. Furthermore, there does not appear to be any conserved amino acid residues used in the AED-specific response other than the framework amino acids. However, when the two clones 8/10-2 and 5/10-20D were compared, a striking similarity was seen in the J segments. These two clones that recognize AED in the context of different MHC epitopes used identical J alpha (J alpha 810) and J beta (J beta 2.6) gene segments. C9, specific for AED-Db, shared identical V beta (V beta 6) and J beta gene segments (J beta 1.1) as those of a cytotoxic T cell that recognizes allogeneic targets expressing Db. These data indicate that a simple rule governing the usage of the variable regions of either the alpha or beta T cell receptor (TcR) genes in the recognition of antigen and MHC gene products cannot be formulated. However, subtle similarities can be detected in some situations between the primary structures of the TcR and the targets they recognize.

Base Sequence↗
PubMed1987

T-cell antigen receptors with identical variable regions but different diversity and joining region gene segments have distinct specificities but cross-reactive idiotypes.

The T-cell antigen receptor alpha-chain genes of an alloreactive, H-2Db-specific cytotoxic T-cell clone (3F9) are described. This study and our work on the 3F9 beta-chain genes reveal that the variable region gene segments for the alpha and beta chains expressed in 3F9 are identical to the ones used by a chicken erythrocyte-specific, I-Ab-restricted helper T-cell clone (LB2). These two clones differ, however, in the diversity and joining portions of the alpha and beta chains of their T-cell receptor molecules. The analysis of 3F9 and LB2 with monoclonal antibodies specific for the 3F9 T-cell receptor shows that these two T-cell clones share the same idiotype; however, 3F9 and LB2 do not exhibit any antigen and/or major histocompatibility complex cross-reactivity. This suggests that the diversity and joining regions of the T-cell receptor may play a key role in antigen and/or major histocompatibility complex recognition.

Animals↗
PubMed1986

T cell-specific gamma genes in C57BL/10 mice. Sequence and expression of new constant and variable region genes.

The T cell-specific gamma genes in C57BL/10 (B10) mice have been analyzed. Based on the cDNA sequences of these genes from antigen-specific MHC-restricted cytotoxic T cells, we found that the repertoire of these genes is not as limited as previously postulated (8). T cells from the B10 mice express an identical copy of V gamma J gamma C gamma (V gamma 10.8A-JC gamma 10.5) transcript previously found in T cells of BALB/c mice. In addition, a potentially functional mRNA using V gamma 10.8B and newly identified J gamma and C gamma gene segments were found. The new J gamma C gamma (JC gamma 10.8) is located 5' to the inverted V gamma 10.8B in the germline DNA of both B10 and BALB/c mice. This new C gamma is only 77 and 66% homologous to the C gamma 10.5 at the nucleotide and deduced protein sequences, respectively, thus making it a potential isotype of the C gamma genes reported previously. The V gamma 5.7, J gamma 2.3 gene segments and pseudogene C gamma 7.5 found in the germline DNA of BALB/c mice are absent in B10 mice. The loss of this gamma chain pseudogene in the B10 mouse strain, and the retention of all potentially functional V gamma, J gamma, and C gamma genes with highly conserved coding sequences supports the importance of these genes.

Amino Acid Sequence↗
PubMedDate not supplied

Roughness induced dynamic changes of wettability of acid etched titanium implant modifications.

Dynamic contact angle analysis (DCA) was used to investigate time-dependent wettability changes of sandblasted and acid-etched commercially pure (cp) titanium (Ti) implant modifications during their initial contact with aqueous systems compared to a macrostructured reference surface. Surface topography was analyzed by scanning electron microscopy and by contact stylus profilometry. The microstructured Ti surfaces were found to be initially extremely hydrophobic. This hydrophobic configuration can shift to a completely wettable surface behavior with water contact angles of 0 degrees after the first emersion loop during DCA experiments. It is suggested that a hierarchically structured surface topography could be responsible for this unexpected wetting phenomenon. Roughness spatial and hybrid parameters could describe topographical features interfering with dynamic wettability significantly better than roughness height parameters. The Ti modifications which shift very sudden from a hydrophobic to a hydrophilic state adsorbed the highest amount of immunologically assayed fibronectin. The results suggest that microstructuring greatly influences both the dynamic wettability of Ti implant surfaces during the initial host contact and the initial biological response of plasma protein adsorption. The microstructured surfaces, once in the totally wettable configuration, may improve the initial contact with host tissue after implantation, due to the drastically increased hydrophilicity.

Acids↗
PubMedDate not supplied

Identical V beta T-cell receptor genes used in alloreactive cytotoxic and antigen plus I-A specific helper T cells.

T lymphocytes involved in the cellular immune response carry cell-surface receptors responsible for antigen and self recognition. This T-cell receptor molecule is a heterodimeric protein consisting of disulphide-linked alpha- and beta-chains with variable (V) and constant (C) regions. Several complementary DNA and genomic DNA clones have been isolated and characterized. These analyses showed that the genomic arrangement and rearrangement of T-cell receptor genes using VT, diversity (DT), joining (JT) and CT gene segments is very similar to the structure of the known immunoglobulin genes. We have isolated two cDNA clones from an allospecific cytotoxic T cell, one of which shows a productive V beta-J beta-C beta 1 rearrangement without an intervening D beta segment. This V beta gene segment is identical to the V beta gene expressed in a helper T-cell clone specific for chicken red blood cells and H-21. The other clone carries the C beta 2 gene of the T-cell receptor, but the C beta 2 sequence is preceded by a DNA sequence that does not show any similarity to V beta or J beta sequences.

Animals↗
PubMedDate not supplied

No functional gamma-chain transcripts detected in an alloreactive cytotoxic T-cell clone.

Three groups of genes that undergo rearrangements during T-cell maturation have been isolated from T cells. Two of them encode the alpha- and beta-subunits of the T-cell antigen receptor and are shared between antigen-specific, major histocompatibility (MHC) class I-restricted cytotoxic T cells and antigen-specific, MHC class II-restricted helper T cells. The third group of genes, called gamma, is preferentially transcribed in cytotoxic T cells. This led to the hypothesis that the unidentified gamma-gene products could be part of a putative T-cell receptor responsible for MHC class I recognition. We report here on the isolation of three different types of gamma-gene transcripts of an alloreactive cytotoxic T-cell clone (3F9). Two are derived from two rearrangements that have occurred at the same locus (V gamma 10.8A to J gamma 10.5 and transcribed with C gamma 10.5), while the third involves a new V gamma-gene segment that is joined to J gamma 13.4 and transcribed with C gamma 13.4. All these rearrangements are abortive and lead to the formation of non-functional gamma-chain genes because the proper translational reading frame is not maintained. Because the second copy of the C gamma 13.4 gene segment is deleted and as C gamma 7.5 is considered to be a pseudogene and has not undergone any rearrangements in 3F9, we conclude that the alloreactive cytotoxic T-cell clone 3F9 does not contain a functional transcript of a known gamma-chain gene.

Animals↗