TCR-γ Gene Rearrangement Research Articles

Are Complicated Forms of Celiac Disease Cryptic T-Cell Lymphomas?

We assessed the clonality of duodenal mucosal T cells in patients with celiac disease and controls. Fifteen adult patients were studied. Four patients had a complicated celiac disease, 3 did not respond to a gluten-free diet, and 2 had an ulcerative jejunitis (including 1 patient with nonresponsive celiac disease). Seven patients had an untreated celiac disease responsive to a gluten-free diet. Histological examination of duodenal biopsies of these 11 patients showed benign-appearing celiac disease without evidence of lymphoma. Four patients with nonulcer dyspepsia and normal duodenal biopsies served as controls. TCRγ gene rearrangements were analyzed by multiplex polymerase chain reaction on DNA extracted from duodenal biopsies. Major clonal rearrangements of the T-cell receptor were found in 4 cases, all with complicated celiac disease. Monoclonality was confirmed by DNA sequencing of the junctional region in 3 cases and by hybridization with clone-specific oligoprobes. Patients with celiac disease responsive to gluten-free diet had mainly a polyclonal pattern, with 1 of them having an oligoclonal rearrangement. An oligoclonal pattern was also observed in 2 control patients. Three patients with complicated celiac disease evolved to T-cell lymphoma with liver (n = 2) or bone marrow (n = 1) invasion. Identical clones were found in the enteropathic duodenojejunum and peripheral blood in the patient with large-cell lymphoma with bone marrow invasion. This study suggests that complicated celiac disease is a cryptic T-cell lymphoma.

Are Complicated Forms of Celiac Disease Cryptic T-Cell Lymphomas?

We assessed the clonality of duodenal mucosal T cells in patients with celiac disease and controls. Fifteen adult patients were studied. Four patients had a complicated celiac disease, 3 did not respond to a gluten-free diet, and 2 had an ulcerative jejunitis (including 1 patient with nonresponsive celiac disease). Seven patients had an untreated celiac disease responsive to a gluten-free diet. Histological examination of duodenal biopsies of these 11 patients showed benign-appearing celiac disease without evidence of lymphoma. Four patients with nonulcer dyspepsia and normal duodenal biopsies served as controls. TCRgamma gene rearrangements were analyzed by multiplex polymerase chain reaction on DNA extracted from duodenal biopsies. Major clonal rearrangements of the T-cell receptor were found in 4 cases, all with complicated celiac disease. Monoclonality was confirmed by DNA sequencing of the junctional region in 3 cases and by hybridization with clone-specific oligoprobes. Patients with celiac disease responsive to gluten-free diet had mainly a polyclonal pattern, with 1 of them having an oligoclonal rearrangement. An oligoclonal pattern was also observed in 2 control patients. Three patients with complicated celiac disease evolved to T-cell lymphoma with liver (n = 2) or bone marrow (n = 1) invasion. Identical clones were found in the enteropathic duodenojejunum and peripheral blood in the patient with large-cell lymphoma with bone marrow invasion. This study suggests that complicated celiac disease is a cryptic T-cell lymphoma.

Alternative splicing of rearranged T cell receptor delta sequences to the constant region of the alpha locus.

The T cell receptor (TCR) alpha/delta locus is composed of a common, shared set of variable (V) and distinct diversity (D), joining (J), and constant (C) genes. It has been recognized for several years that transcripts of the rearranged VDJdelta or VJalpha genes are spliced to the Cdelta or Calpha genes, respectively, encoding distinct TCR delta and alpha proteins. Herein, we describe the discovery of a splicing variation that allows the assembled VDJdelta genes to be fused with the Calpha gene. This variation is prominent in TCRdelta gene-deficient mice but is also detectable in wild-type mice. Furthermore, we show that several in-frame VDJdelta rearrangements in TCRdelta gene-deficient mice are strikingly underrepresented, suggesting that the alternative transcripts, with protein coding capacity, influence the development of alphabeta thymocytes. In-frame TCRgamma gene rearrangements do not appear underrepresented, indicating that the effect is not mediated by the gamma chain. Instead, indirect evidence supports the hypothesis that the delta/alpha chimeric protein acts in conjunction with the TCRbeta chain. These results have implications for the transcriptional control of the TCRalpha/delta locus and provide a novel insight into the distinct functional capacities of the TCR alpha and delta proteins during thymocyte development.

Abnormal intestinal intraepithelial lymphocytes in refractory sprue

Background & Aims: The etiology of refractory sprue is unclear. To gain insight into its pathogenesis, the phenotype and T-cell receptor (TCR) gene rearrangement status of intestinal lymphocytes were analyzed in a group of patients with clinical or biological features of celiac disease but either initially or subsequently refractory to a gluten-free diet. Methods: Intestinal biopsy specimens were obtained from 26 adults: 6 patients with refractory sprue, 7 patients with active celiac disease, and 13 normal controls. The phenotype of intestinal lymphocytes was studied by immunohistochemistry and, in 3 patients with refractory sprue, by cytometry of lymphocytes purified from intestinal biopsy specimens. TCR rearrangements were assessed by studying TCRγV-J junctional regions from DNA extracted from intestinal biopsy specimens and purified intestinal lymphocytes. Results: In the 6 patients with refractory sprue, but not in normal controls or patients with active celiac disease, the intestinal epithelium was massively infiltrated by small lymphocytes that lacked CD8, CD4, and TCR, contained intracytoplasmic but not surface CD3ϵ chains, and exhibited restricted TCRγ gene rearrangements. Conclusions: Refractory sprue is associated with an abnormal subset of intraepithelial lymphocytes containing CD3ϵ and restricted rearrangements of the TCRγ chain but lacking surface expression of T-cell receptors. GASTROENTEROLOGY 1998;114:471-481

Minimal residual disease analysis for the prediction of relapse in children with standard‐risk acute lymphoblastic leukaemia

We report a largely retrospective analysis of minimal residual disease (MRD) in a cohort of 66 children suffering from acute lymphoblastic leukaemia (ALL). All patients lacked high-risk features at diagnosis, i.e. the presenting white cell count was <50 x 10(9)/l, age 1-16 years and translocations t(9;22) and t(4;11) were not present. All were treated according to either the MRC protocols UKALL X or XI. PCR of IgH, TCRdelta and TCRgamma gene rearrangements and allele-specific oligoprobing were employed for the detection of MRD. Sensitivity was at least 10(-4) in 78/82 (93%) probes examined. A total of 33 patients relapsed (seven on therapy and 26 off) and 33 remain in continuing complete remission (CCR) (median follow-up 69 months from diagnosis). Of those who remain in CCR, MRD was present in the bone marrow in 32%, 10% and 0% at 1, 3 and 5 months into therapy respectively. This is in marked contrast to the presence of MRD at these times in 82%, 60% and 41% of patients who relapsed (P<0.001, P<0.005 and P<0.005). These results provide further evidence of a strong correlation between clearance of MRD early in therapy and clinical outcome in childhood ALL.

Granulocyte-colony stimulating factor induces differentiation and apoptosis of CD2, CD7 positive hybrid leukemia cells in vivo and ex vivo

We report a case of a 70-year-old man with a hybrid leukemia treated successfully with granulocyte-colony stimulating factor (G-CSF) combined with a cytocine arabinoside regimen through the induction of differentiation of leukemic cells into monocytoid cells resulting in apoptosis. The leukemic cells demonstrated a TCR-γ gene rearrangement, and expressed CD2, CD7, CD33 and G-CSF receptors but not CD11b on the cell surface nor non-specific esterase in cytoplasm. Several days following the administration of G-CSF, the cells with monocytoid characteristics such as CD11b and cytoplasmic non-specific esterase appeared in the peripheral blood replacing the blastic cells. The cells were shown to be derived from the same clone of the leukemic cell because of the identical TCR-γ gene rearrangement. The short-term culture of leukemic cells with G-CSF induced the differentiation into a monocyte lineage, resulting in apoptosis. Although there is no denying the possibility that cytosine arabinoside is partly responsible, our results strongly suggest that G-CSF plays the main role in differentiation of leukemic cells into a monocyte lineage inducing apoptosis in vivo in this patient.

Defective TCRG gene rearrangement in interleukin-7 receptor knock out mice

Defective TCRG gene rearrangement in interleukin-7 receptor knock out mice

Defective TCRG gene rearrangement in interleukin-7 receptor knock out mice

Defective TCRG gene rearrangement in interleukin-7 receptor knock out mice

Detection of Clonal T-Cell Receptor ? Gene Rearrangements With the Use of the Polymerase Chain Reaction in Cutaneous Lesions of Mycosis Fungoides and Sézary Syndrome

We used the amplification of junctional V (variable)-J joining sequences of the rearranged T-cell receptor gamma (TCR gamma) genes by polymerase chain reaction for rapid and sensitive detection of a clonal T-cell population in a total of 51 skin specimens obtained from 45 patients with mycosis fungoides, five patients with Sézary syndrome, and 29 patients with chronic inflammatory dermatoses. A clonal TCR gamma gene rearrangement was present in all tumors (3/3, 100%) and in most infiltrated plaques (16/22, 73%) and erythrodermas (10/12, 83%). In the patch stage, a clonal subset was found in more than half of the cases (8/14, 57%), whereas no clonality was observed in the controls. We also amplified the V-J sequences of the Igh locus coding for the heavy chain of immunoglobulins, without evidence of clonal rearrangement. These data were compared with those from in situ immunophenotypic analysis. Moreover, by using the same assay with successive dilutions of standard clonal T-cell DNA, a semiquantitative study of the T-cell clone was carried out in some cases. The highest ratios of clonal DNA were observed in advanced stages. These data validate polymerase chain reaction V gamma-J gamma as a rapid, sensitive tool that can be used in the routine analysis of clonality in cutaneous lesions of mycosis fungoides and in the early diagnosis of mycosis fungoides and Sézary syndrome. Semiquantitative studies suggest that the malignant T-cell clone follows a selective process during the course of the progressive form of mycosis fungoides.

Angioimmunoblastic lymphadenopathy with dysproteinemia and dermal T-cell lymphoma

T-cell receptor (TCR)-gamma gene rearrangements provide a specific clonal marker for T-cell malignancies of both the alpha beta and gamma delta varieties. A polymerase chain reaction (PCR)-based method was used in this study for investigation of clonal TCR-gamma gene rearrangements in a patient with a classical presentation of angioimmunoblastic lymphadenopathy with dysproteinemia (AILD) that subsequently progressed into an indolent form of dermal T-cell lymphoma. TCR gene rearrangements in patients with cutaneous T-cell lymphoma (CTCL) were examined using conventional Southern blot analysis and a newly developed PCR-based technique for clonal TCR gene rearrangements. The oligoprimers amplified rearranged V gamma and J gamma segments (including the N region) of the TCR-gamma gene, and PCR products were resolved using high resolution nondenaturing polyacrylamide gel electrophoresis. The authors' results demonstrated good correlation between the two techniques in 10 patients with CTCL (9 patients with C beta and 1 patient with delta 2 rearrangements) and 10 control subjects. The PCR-based technique allowed the authors to detect the presence of an identical T-cell clone in all skin nodules, but not in the original lymph node affected by AILD. This PCR-based method for detecting clonal TCR rearrangements is a highly sensitive and specific technique for detecting T-cell clones in fresh and paraffin embedded tissues. The presence of a T-cell clone in all skin nodules of this patient, but not in the original lymph node affected by AILD, confirms previous findings that in some cases of AILD, clonal T-cell expansion may not be detectable until a later stage of the disease.

Continuing V γ2 to J γ2 rearrangements of murine T cell receptor gamma genes in a B-committed immature cell line

In SPL2-1-2, a murine B-committed immature cell line transformed with a temperature-sensitive mutant of Abelson virus, T cell receptor (TCR) γ gene rearrangements, together with IgH gene rearrangements, were induced during culture at a non-permissive temperature (39°C). During 11–12 months of culture, TCRγ gene rearrangements occurred in all cells. In contrast to TCR γ genes, neither TCR β or TCR δ were detected even after 11–12 months of culture at a non-permissive temperature. The majority of the TCR γ gene rearrangements observed here were V γ 2 to J γ 2 joinings and the remaining rearrangements were J γ 1-linked. V γ 1 to J γ 4 and V γ 3 to Jγ 3 joinings detected. Approximately 70% of cells with TCR γ gene rearrangements produced normal-sized transcripts from the rearranged TCR γ genes. Cloning and sequencing analysis of a cDNA from the transcripts demonstrated that the whole structure of the cDNA was similar to that of T-lineage cells. These results showed that TCR gene rearrangements were restricted to the γ genes and that Vγ 2 to Jγ 2 joinings occurred preferentially in this B-committed immature cell line. Furthermore, TCR γ gene rearrangements also occurred in intracytoplasmic μ -chain producing cells. This indicated that the existence of intracytoplasmic μ-chains did not prevent TCR γ gene rearrangements, although the existence of μ-chains is known to inhibit further productive IgH gene rearrangements, (allelic exclusion). These results should provide many implications for the mechanism of TCR gene rearrangements, especially that of cross-lineage rearrangements.

Ｈｏｄｇｋｉｎ＆Ｒｅｅｄ‐Ｓｔｅｒｎｂｅｒｇ細胞がＨｅｌｐｅｒ／ｉｎｄｕｃｅｒＴ細胞型の表面形質を示したホジキン病の１例

We report a case of Hodgkin's disease showing helper/inducer T cell phenotype on Hodgkin & Reed-Sternberg cells.A 21 years-old Japanese female was admitted for right cervical lymphadenopathy. Laboratory tests including human T lymphocyte virus-I antibody (HTLV-I antibody) were within normal. A biopsy specimen of the cervical lymph node showed Hodgkin's disease, nodular sclerosis (NS).Immunohistologically, Hodgkin & Reed-Sternberg cells were positive for CD30 (Ki-1), CD15 (Leu-M1), CD25 (IL-2R), HLA-DR, transferrin receptor, CD2 (Leu-5b), CD3 (Leu-4), CD4 (Leu-3a), and CD5 (Leu-1), but were negative for B cell associated antigens and myeloid-monocyte associated antigens. The immunohistochemical data indicate helper/inducer T cell phenotype on Hodgkin & Reed-Sternberg cells.Southern blot analysis revealed six rearranged bands using T cell receptor gene probe (TCR Jγ1 probe), when genomic DNA of the biopsied lymph node was digested with EcoRI. Although the TCRγ gene rearrangement bands were not well evaluated, the finding of TCRγ gene rearrangement is unlikely to consider a clonal prolifaration of Hodgkin & Reed-Stenberg cells with T cell phenotype. It would be speculated in this case that rearranged bands for TCRγ may originate from functional T cell clones with transcription of the CD3 molecule and unique TCRγ rearrangements.The patient is free from the disease 2 years after the biopsy.

Trophoblast-derived immunoreguratory factor

The diagnosis of T-cell lymphoproliferative disorders, which frequently involve the skin and other extranodal sites, is often problematic because of the difficulty in establishing clonality in paraffin-embedded tissue. To this end, we developed a simple, nonradioactive method to detect T-cell receptor γ (TCR-γ) gene rearrangements by polymerase chain reaction single-strand conformational polymorphism (PCR-SSCP) in paraffin-embedded tissue. Jurkat and HSB-2 cell lines and peripheral blood samples from normal individuals were used as monoclonal and polyclonal controls, respectively. DNA was extracted from 24 biopsies of T-cell lymphomas, 12 biopsies of reactive lymphoid infiltrates, and 2 biopsies of primary cutaneous large B-cell lymphomas. Vγ1–8, Vγ9, Vγ10, Vγ11, and Jγ1/Jγ2 consensus primers were used for TCR-γ gene rearrangement amplification and PCR products were analyzed by nonradioactive SSCP. Monoclonal controls yielded a well-defined banded pattern, whereas all polyclonal T-cell controls showed a reproducible pattern of smears. We detected monoclonality in 20/21 (95%) T-cell lymphoma cases, whereas no dominant T-cell clones were found in any of the reactive lymphoid infiltrates or B-cell lymphomas. Sensitivity of 1–5% was demonstrated by serially diluting Jurkat cells in mononuclear blood cells from normal individuals. We conclude that nonradioactive PCR-SSCP for TCR-γ gene rearrangement analysis is a useful adjunct to routine histological and immunophenotypic methods in the diagnosis of T-cell lymphoproliferative disorders in paraffin-embedded tissue.