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Structure of the gamma/delta T cell receptor of a human thymocyte clone.

ABSTRACT: The CD3+, IL-2-dependent normal human thymocyte clone, CII, expresses on its surface a CD3-associated gamma/delta TCR. We have further elucidated the structure of this receptor from the nucleotide sequence of cDNA and genomic clones from CII that encode functional TCR-gamma and -delta chains. We find that the CII line expresses a C gamma 2 constant region that is a polymorphic form lacking a copy of an internal exon; the sequence of this constant region accounts for the size of the gamma chain and noncovalent linkage of gamma and delta chains in the CII TCR. The V gamma region used for the CII TCR is identical to the several previously characterized expressed human V gamma segments. Possible implications of this finding are discussed.

SUBMITTER: Okada A

PROVIDER: S-EPMC2189084 | biostudies-other | 1988 Oct

REPOSITORIES: biostudies-other

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Structure of the gamma/delta T cell receptor of a human thymocyte clone.

Okada A A Bank I I Rogozinski L L Takihara Y Y Mak T W TW Chess L L Alt F W FW

The Journal of experimental medicine 19881001 4

The CD3+, IL-2-dependent normal human thymocyte clone, CII, expresses on its surface a CD3-associated gamma/delta TCR. We have further elucidated the structure of this receptor from the nucleotide sequence of cDNA and genomic clones from CII that encode functional TCR-gamma and -delta chains. We find that the CII line expresses a C gamma 2 constant region that is a polymorphic form lacking a copy of an internal exon; the sequence of this constant region accounts for the size of the gamma chain a ...[more]

PMID: 2844954

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Project description:Mechanistic studies contribute greatly to our understanding of γδ T cell (γδTc) biology, aiding development of these cells as immunotherapeutic agents. The antibody blocking assay is an accepted method to determine the receptors involved in γδTc killing of tumor targets. Effectors and/or targets are preincubated with microgram quantities of monoclonal antibodies (mAb), often described by commercial sources to be useful for blocking assays. We and others have used such assays extensively in the past, correlating decreases in cytotoxicity against specific targets with involvement of the blocked receptor(s). However, we wondered whether other mechanisms might be at play beyond cytotoxicity inhibition. Indeed, administration of certain "blocking" mAb to the γδ T cell antigen receptor (γδTCR) induced γδTc death. Upon further investigation, we discovered that γδTc underwent apoptosis triggered by incubation with mAb to the γδTCR. This effect was specific, as no apoptosis was observed when αβ T cells (αβTc) were incubated with these mAb. Apoptosis was further potentiated by the presence of interleukin (IL)-2, often included in cytotoxicity assays; however, exogenous interleukin-2 (IL-2) did not contribute significantly to γδTc cytotoxicity against breast cancer cell lines. Here, we have investigated the usefulness of four mAb for use in blocking assays by assessing blocking properties in conjunction with their propensity to induce apoptosis in cultured primary human γδTc. We found that the 5A6.E9 clone was usually a better alternative to the commonly used B1 (or B1.1) and 11F2 clones; however, some variability in susceptibility to apoptosis induction was observed among donor cultures. Thus, viability assessment of primary effector cells treated with mAb alone should be undertaken in parallel with cytotoxicity assays employing blocking antibodies, to account for cytotoxicity reduction caused by effector cell death. Previous findings should be reassessed in this light.

Project description:BackgroundWC1 co-receptors belong to the scavenger receptor cysteine-rich (SRCR) superfamily and are encoded by a multi-gene family. Expression of particular WC1 genes defines functional subpopulations of WC1(+) γδ T cells. We have previously identified partial or complete genomic sequences for thirteen different WC1 genes through annotation of the bovine genome Btau_3.1 build. We also identified two WC1 cDNA sequences from other cattle that did not correspond to sequences in the Btau_3.1 build. Their absence in the Btau_3.1 build may have reflected gaps in the genome assembly or polymorphisms among animals. Since the response of γδ T cells to bacterial challenge is determined by WC1 gene expression, it was critical to understand whether individual cattle or breeds differ in the number of WC1 genes or display polymorphisms.ResultsReal-time quantitative PCR using DNA from the animal whose genome was sequenced ("Dominette") and sixteen other animals representing ten breeds of cattle, showed that the number of genes coding for WC1 co-receptors is thirteen. The complete coding sequences of those thirteen WC1 genes is presented, including the correction of an error in the WC1-2 gene due to mis-assembly in the Btau_3.1 build. All other cDNA sequences were found to agree with the previous annotation of complete or partial WC1 genes. PCR amplification and sequencing of the most variable N-terminal SRCR domain (domain 1 which has the SRCR "a" pattern) of each of the thirteen WC1 genes showed that the sequences are highly conserved among individuals and breeds. Of 160 sequences of domain 1 from three breeds of cattle, no additional sequences beyond the thirteen described WC1 genes were found. Analysis of the complete WC1 cDNA sequences indicated that the thirteen WC1 genes code for three distinct WC1 molecular forms.ConclusionThe bovine WC1 multi-gene family is composed of thirteen genes coding for three structural forms whose sequences are highly conserved among individual cattle and breeds. The sequence diversity necessary for WC1 genes to function as a multi-genic pattern recognition receptor array is encoded in the genome, rather than generated by recombinatorial diversity or hypermutation.

Dataset Information

Structure of the gamma/delta T cell receptor of a human thymocyte clone.

Publications

Structure of the gamma/delta T cell receptor of a human thymocyte clone.

Similar Datasets

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