Biomedical subjects

J M Dore

Publications and source records attributed to J M Dore.

9 recordsLinked to original sources

PubMed1998

Identification and location on syndecan-1 core protein of the epitopes of B-B2 and B-B4 monoclonal antibodies.

Using a phage display peptide library, we characterized the epitope of two monoclonal antibodies reacting with syndecan-1: B-B2 and B-B4. The identified epitopes QDIT, for B-B2, and LPEV, for B-B4, were found to align with residues 36-39 and 90-93 of the mature protein, respectively. In contrast to B-B4, the B-B2 epitope is close to a potential glycosaminoglycan attachment site. Since syndecan-1 is heavily glycosylated and post-translational modifications are cell type specific, these results might explain the differences observed in the reactivity pattern of B-B2 and B-B4 and suggest that these monoclonal antibodies are useful probes to study cell surface exposed syndecan-1.

Amino Acid Sequence↗

PubMed1997

Expression and activity of a recombinant chimeric protein composed of pokeweed antiviral protein and of human interleukin-2.

The pokeweed antiviral protein (PAP) has already been used to chemically construct immunotoxins. Here we tested the recombinant approach for the production of PAP-containing cytotoxic fusion-proteins. A cDNA encoding a mutated PAP (PAP9), which is expressed at high levels in bacteria, was fused to human interleukin-2 (IL-2) cDNA. The resulting PAP9-IL-2 protein was as active as the free PAP9 in inhibiting an eukaryotic cell-free translation system. Only the chimeric protein desaminated the 28S rRNA and inhibited translation of the CTLL-2 cell line which expresses the IL-2 receptor. These results show that PAP is a suitable toxin for the production of recombinant immunotoxins.

Animals↗

PubMed1996

A plasmocyte selective monoclonal antibody (B-B4) recognizes syndecan-1.

We developed a new monoclonal antibody. B-B4, which specifically identifies human plasma cells. It strongly reacts with all multiple myeloma cell lines and with malignant plasma cells of all tumour samples of the multiple myeloma patients tested. B-B4 does not react with any peripheral blood, bone marrow or tonsil cells. Cloning of the B-B4 antigen reveals that the monoclonal antibody recognizes syndecan-1. It appears that the monoclonal antibody B-B4 is a suitable marker for human plasmocyte identification among haemopoietic cells and a useful probe for the diagnosis of haematological malignancies. Furthermore, this monoclonal antibody can be used for depletions prior to CD34 grafting.

Antibodies, Monoclonal↗

PubMed1993

Mutations dissociating the inhibitory activity of the pokeweed antiviral protein on eukaryote translation and Escherichia coli growth.

The pokeweed antiviral protein is a ribosome inactivating protein acting on eukaryotic as well as on prokaryotic ribosomes thus is toxic for both cell types. Using the PCR technique to clone the PAP open reading frame, we characterized two cDNAs coding for proteins inhibiting eukaryotic translation process and which are not toxic for Escherichia coli, unlike the wild type protein. The sequence of the two cDNAs showed that the proteins contain only one and two point mutations. This result suggest that the wild type amino acids in the mutated positions participate in the prokaryotic ribosome recognition. These mutants might be useful for the construction of immunotoxins containing the pokeweed antiviral protein as toxin.

Amino Acid Sequence↗

PubMed1991

Alfalfa mosaic virus RNA3 mutants do not replicate in transgenic plants expressing RNA3-specific genes.

The RNA3 of alfalfa mosaic virus (AIMV) encodes the P3 protein and the viral coat protein (CP). RNA3 molecules transcribed in vitro replicated in protoplasts and plants when inoculated in mixtures with AIMV RNA1, RNA2 and CP. Transcripts with a deletion or inversion in the P3 gene replicated well in protoplasts but not in transgenic plants transformed with the P3 gene. Transgenic plants expressing the CP gene became infected after inoculation with a mixture of RNA1, RNA2 and wild-type RNA3 transcripts without addition of CP to the inoculum. Transcripts with a deletion in the CP gene replicated at a reduced level in protoplasts but not in CP-transformed plants. This suggests that P3 and CP are both required for cell-to-cell spread of AIMV and that mutations in the inoculum RNA could not be complemented in trans by the wild-type chimeric nuclear genes.

Blotting, Northern↗

PubMed1991

Identification of potyviruses using the polymerase chain reaction with degenerate primers.

Local areas of conserved amino acid sequence in the replicase and coat proteins of potyviruses were used to select nucleotide sequences for use in the construction of sets of degenerate oligonucleotide primers for amplification of DNA fragments on potyvirus-specific templates in a combined assay of reverse transcription and the polymerase chain reaction (RT-PCR). Sequences selected for the construction of degenerate primers included the coat protein gene sequence of tulip breaking virus from lily, which is reported in this paper. It is shown that the degenerate primers support potyvirus-specific amplification, but do not support amplification on carlavirus and potexvirus templates. A panel consisting of definite and prospective members of the potyvirus group occurring in bulbous crops was subjected to the degenerate primer RT-PCR assay; amplified fragments were used in cross-hybridization experiments and restriction fragment length polymorphism analysis to detect relationships among these potyviruses. A partially characterized virus isolated from Gloriosa rothschildiana was positively identified as a potyvirus by specific amplification and subsequent sequence analysis of an amplified DNA fragment.

Amino Acid Sequence↗

PubMed1990

Biologically active transcripts of alfalfa mosaic virus RNA3.

Transcripts of the bicistronic RNA3 of alfalfa mosaic virus were synthesized using the in vitro T7 run-off transcription system. Synthetic RNA3 containing one additional G nucleotide at the 5' end were found to be infectious when coinoculated with RNA1 and RNA2 and coat protein.

Base Sequence↗

PubMed1989

Competitive multiplication of RNA3 species of different strains of alfalfa mosaic virus.

Competition between RNA3 from alfalfa mosaic virus (AlMV) strain S (RNA3-S), strain B (RNA3-B) and strain 425L (RNA3-L) was studied. The identification of the RNA3 species multiplying in infected leaves was possible since the RNA3 5' non-coding leader sequences in strains S, B and 425L differ in length. RNA3 present in total RNA from infected tobacco leaves was detected, and strains were identified from the length of the cDNA reverse-transcribed from RNA primed with a specific oligonucleotide. In competition experiments the inoculum, containing known amounts of RNA1, 2, 3 and 4 of one strain, was complemented with various amounts of heterologous RNA3 and inoculated to a systemic host. It is shown that RNA3-S was better replicated in vivo by the AlMV replicase of strain B than was RNA3-B itself, and to a lesser extent better replicated by the AlMV replicase of strain L than was RNA-L. Comparison of genetic information carried by the RNA3 species present in the inoculum suggests that the more efficient multiplication of RNA3-S is related to the structure of the leader sequence of RNA3-S.

Amino Acid Sequence↗

PubMedDate not supplied

CD138.

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Amino Acid Sequence↗