Linked Life Data

8576577

Source:http://linkedlifedata.com/resource/pubmed/id/8576577

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
1996-3-13
pubmed:abstractText
We recently showed that secretion of non-chimeric disulfide-linked human gamma delta TCR ('soluble' TCR, sTCR) comprising V gamma 9 and V delta 2 regions could be achieved by simply introducing translational termination codons upstream from the sequences encoding TCR transmembrane region. Here we extended these findings by demonstrating efficient secretion and heterodimerization of gamma delta sTCR comprising V gamma 8, V delta 1 and V delta 3 regions, obtained via the same strategy. After immunization against immunoaffinity-purified soluble TCR, several hundreds of TCR-specific monoclonal antibodies (mAb) were generated, which fell in at least seven groups. One set of mAb was directed against a V gamma 8-specific epitope. Strikingly, despite the high degree of sequence homology between V gamma 8 and other V gamma I domains, none of these mAb were crossreactive with other members of the V gamma I family. Three other sets of mAbs were shown to recognize delta chains comprising V delta 1, V delta 2 and V delta 3 regions respectively, regardless of their junctional sequence or of the gamma chain to which they were paired. Among the V delta 1-specific mAb, some specifically recognized V delta 1D delta J delta C delta chains while others reacted with both V delta 1 D delta J delta C delta and V delta 1J alpha C alpha chains, which suggested V domain conformational alterations induced by the C region. Moreover, reactivity of one V delta 1-specific mAb (#R6.11) was affected by a polymorphic residue located on the predicted CDR4 loop of the V delta region. Two delta chain-specific mAb (#178 and #515) showed a highly unusual reactivity, which was negatively affected by particular V delta and J delta sequences: (i) mAb #515 and #178 recognized all TCR delta chains except those comprising V delta 1 or V delta 2 regions, respectively and (ii) within TCR delta chains carrying 'permissive' V delta regions, none of those comprising the J delta 2 region were recognized by #515 and/or #178 mAbs, which suggested a highly specific conformation adopted by this particular J delta sequence. Apart from its usefulness in TCR structural studies, this novel set of mAb represents an important tool for the characterization and isolation of gamma delta T cells expressing particular combinations of V gamma/V delta regions and for analysis of V alpha/V delta usage by alpha beta T cells. Moreover, since our present data strongly suggest that gamma delta TCR are easier to obtain in a soluble form than alpha beta TCR, an efficient strategy for the generation of V alpha region-specific mAb might be to immunize with chimeric gamma delta sTCR comprising particular V alpha regions.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jan
pubmed:issn
0022-1759
pubmed:author
pubmed:issnType
Print
pubmed:day
16
pubmed:volume
189
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
25-36
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:8576577-Amino Acid Sequence, pubmed-meshheading:8576577-Animals, pubmed-meshheading:8576577-Antibodies, Monoclonal, pubmed-meshheading:8576577-Antibody Specificity, pubmed-meshheading:8576577-Base Sequence, pubmed-meshheading:8576577-CHO Cells, pubmed-meshheading:8576577-Cricetinae, pubmed-meshheading:8576577-Electrophoresis, Polyacrylamide Gel, pubmed-meshheading:8576577-Humans, pubmed-meshheading:8576577-Immunization, pubmed-meshheading:8576577-Mice, pubmed-meshheading:8576577-Mice, Inbred BALB C, pubmed-meshheading:8576577-Molecular Sequence Data, pubmed-meshheading:8576577-Polymorphism, Genetic, pubmed-meshheading:8576577-Protein Conformation, pubmed-meshheading:8576577-Protein Engineering, pubmed-meshheading:8576577-Receptors, Antigen, T-Cell, gamma-delta, pubmed-meshheading:8576577-Recombinant Fusion Proteins
pubmed:year
1996
pubmed:articleTitle
Structural analysis of gamma delta TCR using a novel set of TCR gamma and delta chain-specific monoclonal antibodies generated against soluble gamma delta TCR. Evidence for a specific conformation adopted by the J delta 2 region and for a V delta 1 polymorphism.
pubmed:affiliation
Immunotech SA, Marseilles, France.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't