Configuration and expression of the T cell receptor beta chain gene in human T-lymphotrophic virus I-infected cells.

We studied the configuration and expression of the gene encoding the beta chain of the T cell receptor (TCR beta) in cell lines and primary tumor cells infected by the human T cell leukemia/lymphoma (lymphotrophic) virus type I (HTLV-I). Most of the cell lines and all the primary tumor cells showed rearrangement of the TCR beta gene, and in each case the rearrangement was distinct. The majority of cases examined were clonal with respect to a particular TCR beta gene rearrangement. Primary tumor cells from one case (SD) were found to have a tandem duplication of a portion of chromosome 7; this appears to have resulted in the presence of three alleles of the TCR beta gene, each of which is arranged differently. This suggests that the chromosomal abnormality, and possibly infection by HTLV-I, occurred before TCR beta gene rearrangement. Cell lines infected by HTLV-I express levels of TCR beta mRNA similar to PHA stimulated lymphocytes, suggesting that this gene is not transcriptionally activated as a result of infection by HTLV-I. Cloned T cells of known antigen specificity that are infected by HTLV-I in vitro show impairment of immune function, including loss of antigen-specific responsiveness and the acquisition of alloreactivity. Comparison of the configuration of the TCR beta gene before and after infection revealed no changes detectable by Southern blot analysis. Levels of expression of the TCR beta gene at the mRNA level and surface expression of the T3 complex were also not significantly altered, suggesting that changes in immune function cannot be attributed to quantitative changes in the TCR molecule. The configuration of the TCR beta gene in primary tumor cells infected by HTLV-I was compared with that in the derived cell lines. In all pairs examined, the configuration in the primary tumor cells was different from that in the cell lines, strongly suggesting that the cells that grow in culture are not the original neoplastic cells.

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Rearrangement of the T cell receptor gamma-chain gene in childhood acute lymphoblastic leukemia

Blood ◽

10.1182/blood.v70.6.1933.bloodjournal7061933 ◽

1987 ◽

Vol 70 (6) ◽

pp. 1933-1939

Author(s):

A Tawa ◽

SH Benedict ◽

J Hara ◽

N Hozumi ◽

EW Gelfand

Keyword(s):

Acute Lymphoblastic Leukemia ◽

T Cell ◽

T Cell Receptor ◽

Gene Rearrangement ◽

Lymphoblastic Leukemia ◽

Leukemia Cell ◽

Cell Receptor ◽

Gene Rearrangements ◽

Gamma Chain ◽

Beta Gene

We analyzed rearrangements of the T cell receptor gamma-chain (T gamma) gene as well as rearrangements of the T cell receptor beta-chain (T beta) gene and immunoglobulin heavy-chain (IgH) gene in 68 children with acute lymphoblastic leukemia (ALL). All 15 patients with T cell ALL showed rearrangements of both T beta and T gamma genes. Twenty-four of 53 non-T, non-B ALL patients (45%) showed T gamma gene rearrangements and only 14 of these also showed T beta gene rearrangements. Only a single patient rearranged the T beta gene in the absence of T gamma gene rearrangement. The rearrangement patterns of the T gamma gene in non-T, non-B ALL were quite different from those observed in T cell ALL, as 20 of 23 patients retained at least one germline band of the T gamma gene. In contrast, all T cell ALL patients showed no retention of germline bands. These data indicate that rearrangement of the T gamma gene is not specific for T cell ALL. Further, the results also suggest that T gamma gene rearrangement precedes T beta gene rearrangement. The combined analysis of rearrangement patterns of IgH, T beta, and T gamma genes provides new criteria for defining the cellular origin of leukemic cells and for further delineation of leukemia cell heterogeneity.

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T-cell receptor beta-chain gene rearrangement and expression during human thymic ontogenesis

Blood ◽

10.1182/blood.v79.6.1472.1472 ◽

1992 ◽

Vol 79 (6) ◽

pp. 1472-1483 ◽

Cited By ~ 19

Author(s):

A Bonati ◽

P Zanelli ◽

S Ferrari ◽

A Plebani ◽

B Starcich ◽

...

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Cell Receptor ◽

Chain Gene ◽

Gene Transcript ◽

Beta Chain ◽

Early Stages ◽

Human Thymus ◽

Nucleotide Insertions ◽

Beta Gene

Abstract T-cell receptor (TCR) beta-chain proteins appear early (approximately 15th week of gestation) during human thymic ontogenesis. These beta- chain proteins, which appear before terminal deoxynucleotidyl transferase (TdT), could be an expression of a fully rearranged (V-D- J), incompletely rearranged (D-J), or germline TCR beta-chain gene. The aims of this study, performed from the 15th week onward, were the following: (1) to investigate whether or not TCR beta gene rearranges at an early stage during human thymic ontogenesis; (2) to investigate whether complete presumptive functional (1.3 kb) TCR beta gene transcript is present at these early stages of development, or if incomplete (1 kb) or germ-line (1.1 kb) transcripts are expressed; (3) to examine the phenotype of TCR beta-chain+ cells with two-color fluorescence using monoclonal antibody (MoAb) beta F1 and MoAbs that recognize CD1, CD2, CD3, CD4, CD8, CD5, and CD7 antigens (rabbit anti- calf TdT antiserum was used to detect TdT); and (4) to demonstrate whether or not beta gene N-diversity regions are detectable as early as the 15th week and whether or not N-nucleotide insertions correlate to TdT expression. Fifteen- to 22-week fetal thymuses and pediatric thymuses were investigated. We demonstrated that TCR beta-chain gene rearranged as early as the 15th week in human thymus and that a complete functional TCR beta gene transcript was expressed at these early stages of human development. No other analyses to date have investigated TCR beta gene expression in early human thymus using molecular biology techniques. No significant differences were detectable between phenotypic analysis of fetal and pediatric samples, except for TdT expression, which appeared after the 20th week. Essentially all mCD3+ (OKT3+) cells were beta-chain+ at the different weeks investigated. A significant percentage of CD1+ cells were beta- chain+, and the percentage increased along with the age of development. After the 20th week, we identified three main populations: TdT+, cCD3+, beta F-(early thymic precursors); TdT+, CD1+, beta F1+ (intermediate maturity cortical thymocytes); and TdT-, mCD3+, beta F1++ (mature medullary thymocytes). Given these values, we may consider beta-chain expression an ordered process. beta gene N-nucleotide insertions were correlated to TdT expression, since N-regions increased considerably after the 20th week. A further increase of N-nucleotide insertions was detected from the 22nd week to the 32nd week.

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Similar rearrangements of T-cell receptor beta gene in cell lines and uncultured cells from patients with large granular lymphocyte leukemia

Blood ◽

10.1182/blood.v72.2.613.613 ◽

1988 ◽

Vol 72 (2) ◽

pp. 613-615 ◽

Cited By ~ 1

Author(s):

TP Jr Loughran ◽

G Starkebaum ◽

FW Ruscetti

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Cell Lines ◽

Interleukin 2 ◽

Cell Receptor ◽

Large Granular Lymphocyte ◽

Phenotypic Analysis ◽

T Cell Receptor Beta ◽

Beta Gene ◽

Receptor Beta

Abstract We established interleukin-2-(IL-2) dependent cell lines from three patients with large granular lymphocyte (LGL) leukemia. Phenotypic analysis demonstrated retention of the CD3+, CD8+ phenotype that was observed in the original leukemic LGL. Unique rearrangements of T-cell receptor beta gene occurring in uncultured leukemic LGL, were also found in cell lines, which suggests that the cell lines were derived from the original leukemic LGL clone in each case.

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Generation of diversity in T cell receptor repertoire specific for pigeon cytochrome c.

Journal of Experimental Medicine ◽

10.1084/jem.165.2.279 ◽

1987 ◽

Vol 165 (2) ◽

pp. 279-301 ◽

Cited By ~ 63

Author(s):

S B Sorger ◽

S M Hedrick ◽

P J Fink ◽

M A Bookman ◽

L A Matis

Keyword(s):

T Cell ◽

Cytochrome C ◽

T Cell Receptor ◽

Cell Lines ◽

Dna Sequences ◽

Cell Receptor ◽

T Cell Response ◽

Beta Chain ◽

Antigen Specificity ◽

T Cell Lines

17 T cell clones and 3 T cell lines, specific for pigeon cytochrome c, were analyzed for fine specificity and rearranged T cell receptor (TCR) gene elements. Clones of similar fine specificities were grouped into one of four phenotypes, and correlations between phenotype differences and gene usage could be made. All the lines and clones rearranged a member of the V alpha 2B4 gene family to a limited number of J alpha regions. The beta chain was made up of one of three non-cross-hybridizing V beta regions, each rearranging to only one or two J beta s. The use of alternate V beta regions could be correlated with phenotype differences, which were manifested either as MHC- or MHC and antigen-specificity changes. In addition, the presence of alloreactivity, which defined a phenotype difference, could be correlated solely with the use of an alternate J alpha region. These observations were substantiated by prospective analyses of pigeon cytochrome c-specific T cell lines that were selected for alternate MHC specificity or alloreactivity and were found to express the correlated alpha and beta chain rearrangements. Previously, the TCR DNA sequences from two clones, each representing a variant of one phenotype, showed sequence differences only in the N regions of their TCR genes. Since only these two variants, using identical V alpha-J alpha and V beta-J beta gene elements, were repeatedly observed in this study, we would predict that the junctional diversity differences are selectable. In this T cell response, all the gene elements involved in the generation of diversity appear to be selected, and may therefore be important in the determination of TCR specificity. This high degree of receptor gene selection represents a fundamental difference from the diversity seen in several extensively analyzed antibody responses.

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Regulation of pre-T cell receptor (pT alpha-TCR beta) gene expression during human thymic development.

Journal of Experimental Medicine ◽

10.1084/jem.184.2.519 ◽

1996 ◽

Vol 184 (2) ◽

pp. 519-530 ◽

Cited By ~ 51

Author(s):

A R Ramiro ◽

C Trigueros ◽

C Márquez ◽

J L San Millán ◽

M L Toribio

Keyword(s):

T Cell ◽

T Cell Receptor ◽

T Cell Development ◽

Cell Receptor ◽

Beta Chain ◽

Thymocyte Development ◽

Gene Encoding ◽

Thymic Development ◽

Alpha Beta ◽

Beta Gene

In murine T cell development, early thymocytes that productively rearrange the T cell receptor (TCR) beta locus are selected to continue maturation, before TCR alpha expression, by means of a pre-TCR alpha- (pT alpha-) TCR beta heterodimer (pre-TCR). The aim of this study was to identify equivalent stages in human thymocyte development. We show here that variable-diversity-joining region TCR beta rearrangement and the expression of full-length TCR beta transcripts have been initiated in some immature thymocytes at the TCR alpha/beta- CD4+CD8- stage, and become common in a downstream subset of TCR alpha/beta- CD4+CD8+ thymocytes that is highly enriched in large cycling cells. TCR beta chain expression was hardly detected in TCR alpha/beta- CD4+CD8- thymocytes, whereas cytoplasmic TCR beta chain was found in virtually all TCR alpha/beta- CD4+CD8+ blasts. In addition, a TCR beta complex distinct from the mature TCR alpha/beta heterodimer was immunoprecipitated only from the latter subset. cDNA derived from TCR alpha/beta- CD4+CD8+ blasts allowed us to identify and clone the gene encoding the human pT alpha chain, and to examine its expression at different stages of thymocyte development. Our results show that high pT alpha transcription occurs only in CD4+CD8- and CD4+CD8+ TCR alpha/beta- thymocytes, whereas it is weaker in earlier and later stages of development. Based on these results, we propose that the transition from TCR alpha/beta- CD4+CD8- to TCR alpha/beta- CD4+CD8+ thymocytes represents a critical developmental stage at which the successful expression of TCR beta promotes the clonal expansion and further maturation of human thymocytes, independent of TCR alpha.

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Similar rearrangements of T-cell receptor beta gene in cell lines and uncultured cells from patients with large granular lymphocyte leukemia

Blood ◽

10.1182/blood.v72.2.613.bloodjournal722613 ◽

1988 ◽

Vol 72 (2) ◽

pp. 613-615 ◽

Cited By ~ 1

Author(s):

TP Jr Loughran ◽

G Starkebaum ◽

FW Ruscetti

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Cell Lines ◽

Interleukin 2 ◽

Cell Receptor ◽

Large Granular Lymphocyte ◽

Phenotypic Analysis ◽

T Cell Receptor Beta ◽

Beta Gene ◽

Receptor Beta

We established interleukin-2-(IL-2) dependent cell lines from three patients with large granular lymphocyte (LGL) leukemia. Phenotypic analysis demonstrated retention of the CD3+, CD8+ phenotype that was observed in the original leukemic LGL. Unique rearrangements of T-cell receptor beta gene occurring in uncultured leukemic LGL, were also found in cell lines, which suggests that the cell lines were derived from the original leukemic LGL clone in each case.

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Validation of Sixteen Leukemia and Lymphoma Cell Lines as Controls for Molecular Gene Rearrangement Assays

Clinical Chemistry ◽

10.1093/clinchem/48.8.1344 ◽

2002 ◽

Vol 48 (8) ◽

pp. 1344-1351 ◽

Cited By ~ 3

Author(s):

Rong Yao ◽

Steven A Rich ◽

Erasmus Schneider

Keyword(s):

T Cell ◽

Southern Blot ◽

T Cell Receptor ◽

Cell Lines ◽

Gene Rearrangement ◽

Cell Receptor ◽

Lymphoma Cell ◽

Clinical Samples ◽

Gene Rearrangements ◽

Positive Controls

Abstract Background: Assays for rearrangement of the immunoglobulin, T-cell receptor, bcr/abl, and bcl-2 genes are valuable tools to aid in the diagnosis of leukemias and lymphomas and are now offered by many pathology laboratories. However, there is a lack of well-characterized and validated calibrators and positive controls for these assays. We therefore evaluated 16 readily available leukemia and lymphoma cell lines for their potential use as controls. Methods: DNA and RNA were isolated from each cell line and analyzed by Southern blot and PCR or reverse transcription-PCR (RT-PCR). Rearrangements in the IgJH, IgJκ, TcR-β or TcR-γ, bcr/abl, and bcl-2 genes were detected by commercially available probes and primers. Cell lineages were confirmed by immunophenotyping. Results: Immunoglobulin and T-cell receptor gene rearrangements were identified in five B- and three T-cell lines, respectively. Two cell lines tested positive for the bcr/abl gene, and one was positive for the bcl-2 gene rearrangement by Southern blot. Conclusions: The 16 cell lines studied can be used as positive controls in molecular detection assays for gene rearrangements. The parallel processing of these cell lines with clinical samples can serve to quality control the experimental procedures from the first step of DNA or RNA extraction to the final step of result analysis.

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T-cell receptor beta-chain gene rearrangement and expression during human thymic ontogenesis

Blood ◽

10.1182/blood.v79.6.1472.bloodjournal7961472 ◽

1992 ◽

Vol 79 (6) ◽

pp. 1472-1483 ◽

Cited By ~ 1

Author(s):

A Bonati ◽

P Zanelli ◽

S Ferrari ◽

A Plebani ◽

B Starcich ◽

...

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Cell Receptor ◽

Chain Gene ◽

Gene Transcript ◽

Beta Chain ◽

Early Stages ◽

Human Thymus ◽

Nucleotide Insertions ◽

Beta Gene

T-cell receptor (TCR) beta-chain proteins appear early (approximately 15th week of gestation) during human thymic ontogenesis. These beta- chain proteins, which appear before terminal deoxynucleotidyl transferase (TdT), could be an expression of a fully rearranged (V-D- J), incompletely rearranged (D-J), or germline TCR beta-chain gene. The aims of this study, performed from the 15th week onward, were the following: (1) to investigate whether or not TCR beta gene rearranges at an early stage during human thymic ontogenesis; (2) to investigate whether complete presumptive functional (1.3 kb) TCR beta gene transcript is present at these early stages of development, or if incomplete (1 kb) or germ-line (1.1 kb) transcripts are expressed; (3) to examine the phenotype of TCR beta-chain+ cells with two-color fluorescence using monoclonal antibody (MoAb) beta F1 and MoAbs that recognize CD1, CD2, CD3, CD4, CD8, CD5, and CD7 antigens (rabbit anti- calf TdT antiserum was used to detect TdT); and (4) to demonstrate whether or not beta gene N-diversity regions are detectable as early as the 15th week and whether or not N-nucleotide insertions correlate to TdT expression. Fifteen- to 22-week fetal thymuses and pediatric thymuses were investigated. We demonstrated that TCR beta-chain gene rearranged as early as the 15th week in human thymus and that a complete functional TCR beta gene transcript was expressed at these early stages of human development. No other analyses to date have investigated TCR beta gene expression in early human thymus using molecular biology techniques. No significant differences were detectable between phenotypic analysis of fetal and pediatric samples, except for TdT expression, which appeared after the 20th week. Essentially all mCD3+ (OKT3+) cells were beta-chain+ at the different weeks investigated. A significant percentage of CD1+ cells were beta- chain+, and the percentage increased along with the age of development. After the 20th week, we identified three main populations: TdT+, cCD3+, beta F-(early thymic precursors); TdT+, CD1+, beta F1+ (intermediate maturity cortical thymocytes); and TdT-, mCD3+, beta F1++ (mature medullary thymocytes). Given these values, we may consider beta-chain expression an ordered process. beta gene N-nucleotide insertions were correlated to TdT expression, since N-regions increased considerably after the 20th week. A further increase of N-nucleotide insertions was detected from the 22nd week to the 32nd week.

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Oligoclonal expansion of CD8+ CD57+ T cells with restricted T-cell receptor beta chain variability after bone marrow transplantation

Blood ◽

10.1182/blood.v83.2.587.bloodjournal832587 ◽

1994 ◽

Vol 83 (2) ◽

pp. 587-595 ◽

Cited By ~ 1

Author(s):

G Gorochov ◽

P Debre ◽

V Leblond ◽

B Sadat-Sowti ◽

F Sigaux ◽

...

Keyword(s):

Bone Marrow ◽

T Cell ◽

T Cell Receptor ◽

Gene Families ◽

Cell Receptor ◽

Pcr Analysis ◽

Gene Rearrangements ◽

Beta Chain ◽

Beta Gene ◽

Oligoclonal Expansion

A major expansion of CD8+57+ lymphocytes expressing an alpha-beta T- cell receptor (TCR) is frequent after bone marrow transplantation (BMT). We examined the clonality of the TCR beta gene repertoire in these expanded CD8+57+ cells after allogeneic or autologous BMT. We performed a polymerase chain reaction (PCR) analysis of the V beta chain usage in CD8+57+ cells purified from nine BMT recipients with a series of oligonucleotides specific for 20 V beta gene families. PCR products from selected TCR beta gene rearrangements were sequenced. The CD8+57+ cells from eight of nine patients used a restricted set of V beta families, with a marked predominance of two to three V beta gene families per patient, whereas the control autologous CD57- subset expressed the whole 20 V beta families. A direct sequencing analysis confirmed the V beta 16 and V beta 17 clonality in six patients, showing a striking homology in the CDR3 sequences of the V beta 16 products. The CD8+57+ cells, but not the CD57- cells, displayed an oligoclonal pattern of TCR rearrangements as shown by PCR analysis of TCR gamma gene rearrangements. Such an oligoclonal expansion of CD8+57+ cells, using a restricted set of the V beta gene families, may result from a specific TCR stimulation of a limited number of T-cell clones in BMT recipients.

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