Cell Receptor Beta Chain Gene Research Articles

Establishment and characterization of a cell line, TOM-1, derived from a patient with Philadelphia chromosome-positive acute lymphocytic leukemia

A new Philadelphia chromosome (Ph1)-positive cell line, designated TOM- 1, was derived from bone marrow cells of a patient with Ph1-positive acute lymphocytic leukemia (ALL). The TOM-1 cells were positive for Ia and B1 antigens and terminal deoxynucleotidyl transferase (TdT) but negative for common ALL antigen. Although neither surface Ig nor cytoplasmic Ig was detected, the TOM-1 cells contained rearranged immunoglobulin-H chain genes but retained germ-line kappa chain and germ-line T cell receptor beta-chain genes. These results indicate that the TOM-1 cells reside as the progenitor of pre-B cells. We have investigated the chromosome 22 breakpoint and c-abl gene expression in the TOM-1 cells. We found that the breakpoint on chromosome 22 was within the breakpoint cluster region (bcr) in the TOM-1 cells. We also found the breakpoints within or near bcr in four of six Ph1-positive ALL cases, similar to the findings in Ph1-positive CML cases. Amplification of the c-abl gene was not detected in the TOM-1 cells. The leukemic cells isolated from a patient with CML in myeloid crisis contained a novel 8-kilobase (kb) abl-related messenger RNA (mRNA), but the TOM-1 cells contained c-abl transcripts of only normal sizes, despite the fact that they showed the bcr gene rearrangement.

Establishment and Characterization of a Cell Line, TOM-1, Derived From a Patient With Philadelphia Chromosome-Positive Acute Lymphocytic Leukemia

A new Philadelphia chromosome (Ph1)-positive cell line, designated TOM-1, was derived from bone marrow cells of a patient with Ph1-positive acute lymphocytic leukemia (ALL). The TOM-1 cells were positive for Ia and B1 antigens and terminal deoxynucleotidyl transferase (TdT) but negative for common ALL antigen. Although neither surface Ig nor cytoplasmic Ig was detected, the TOM-1 cells contained rearranged immunoglobulin-H chain genes but retained germ-line kappa chain and germ-line T cell receptor beta-chain genes. These results indicate that the TOM-1 cells reside as the progenitor of pre-B cells. We have investigated the chromosome 22 breakpoint and c-abl gene expression in the TOM-1 cells. We found that the breakpoint on chromosome 22 was within the breakpoint cluster region (bcr) in the TOM-1 cells. We also found the breakpoints within or near bcr in four of six Ph1-positive ALL cases, similar to the findings in Ph1-positive CML cases. Amplification of the c-abl gene was not detected in the TOM-1 cells. The leukemic cells isolated from a patient with CML in myeloid crisis contained a novel 8-kilobase (kb) abl-related messenger RNA (mRNA), but the TOM-1 cells contained c-abl transcripts of only normal sizes, despite the fact that they showed the bcr gene rearrangement.

T cell gamma gene rearrangements in hematologic neoplasms

Rearrangements of the T cell gamma (T gamma) gene were studied in primary neoplastic cells from 75 patients with leukemia or lymphoma. T gamma gene rearrangements were observed in 19 of 21 T cell neoplasms; 14 of 21 immature B cell leukemias, including 4 out of 5 patients with rearrangements of both immunoglobulin heavy-chain (JH) and T cell receptor beta chain (T beta) genes; none out of 16 nonlymphoid leukemias. Thus, T gamma gene rearrangement is frequently found in immature B cells and is not always found in T cells showing T beta gene rearrangement, but it is not detected in nonlymphoid cells. Furthermore, T gamma gene rearrangement in cells with the germline configuration of the JH and T beta genes was observed. These results indicate that the detection of T gamma gene rearrangement does not allow a clear assignment to a particular lineage. However, an analysis of T gamma gene rearrangement provides a further potential tool to establish the lymphoid cellular origin and clonality of hematologic neoplasms and identify the normal stages of lymphocyte differentiation.

T Cell γ Gene Rearrangements in Hematologic Neoplasms

Rearrangements of the T cell gamma (T gamma) gene were studied in primary neoplastic cells from 75 patients with leukemia or lymphoma. T gamma gene rearrangements were observed in 19 of 21 T cell neoplasms; 14 of 21 immature B cell leukemias, including 4 out of 5 patients with rearrangements of both immunoglobulin heavy-chain (JH) and T cell receptor beta chain (T beta) genes; none out of 16 nonlymphoid leukemias. Thus, T gamma gene rearrangement is frequently found in immature B cells and is not always found in T cells showing T beta gene rearrangement, but it is not detected in nonlymphoid cells. Furthermore, T gamma gene rearrangement in cells with the germline configuration of the JH and T beta genes was observed. These results indicate that the detection of T gamma gene rearrangement does not allow a clear assignment to a particular lineage. However, an analysis of T gamma gene rearrangement provides a further potential tool to establish the lymphoid cellular origin and clonality of hematologic neoplasms and identify the normal stages of lymphocyte differentiation.

T cell receptor β chain gene rearrangements and Vβ gene usage in horse cytochrome c‐specific T cell hybridomas

Comparison of T cell receptor alpha and beta-chain genes in murine major histocompatibility complex (MHC) class I and class II-restricted T cell clones and hybridomas recognizing different antigens indicates that no simple correlation exists between the observed antigen/MHC specificity and the expression of certain alpha and beta-chain heterodimers. We have attempted to establish a possible correlation by analyzing T cell receptor beta chain gene rearrangements and V beta gene usage in five T cell hybridomas with identical antigen/MHC specificity and another hybridoma recognizing a different antigenic determinant in association with the same restriction molecule. We report here that in each of the five clones a uniquely rearranged beta chain gene is expressed in combination with at least two different V beta gene segments. The presence of the differently rearranged T cell receptor beta chain genes correlated with the finding of distinct fine specificity pattern of antigen recognition in each of the hybridomas. Interestingly, two hybridomas specific for different epitopes showed identical beta chain D-J rearrangements indicating that the differences might be encoded by the alpha chain gene or/and the V beta gene element.

7q32-q36 translocations in childhood T cell leukemia: cytogenetic evidence for involvement of the T cell receptor beta-chain gene

Blast cell chromosomal rearrangements involving the long arm of chromosome 7 were identified in eight of 197 cases of childhood acute lymphoblastic leukemia (ALL). Breakpoints were variable but tended to cluster in either the proximal or the terminal 7q region, depending on the immunophenotype of the cells. The 7q32–q36 region, the locus of the T cell receptor beta-chain gene, was the site of breakpoints in four of 31 cases of T cell ALL but was not involved in any of the 166 cases originating from B cell precursors (P less than .0004). In three of the four T cell cases it was possible to identify the chromosomal segment that had been translocated to the 7q32-q36 region: 1p32, 2p21, and 6p21. The 1p32 and 6p21 bands are particularly interesting, as they contain the sites of two known protooncogenes, c-L-myc and hpim, respectively. Our findings suggest that the locus of the beta-chain gene of the T cell receptor is a preferential site for certain chromosomal rearrangements in leukemic T lymphoblasts, analogous to the T cell receptor alpha-chain gene on human chromosome 14. Translocation of proto-oncogenes to a site near the beta-chain regulatory sequences provides a potential mechanism for oncogene activation.

T Cell Receptor Beta Chain Gene Polymorphisms are Associated with Insulin Dependent Diabetes in Identical Twins

T Cell Receptor Beta Chain Gene Polymorphisms are Associated with Insulin Dependent Diabetes in Identical Twins

Immunoprecipitation of the interleukin‐2 receptor from Hodgkin's disease derived cell lines by monoclonal antibodies

The nature of Hodgkin and Reed-Sternberg (H-RS) cells and their normal counterpart remains a matter of controversy. Our recent investigations have suggested that H-RS-cells derive from certain activated lymphocytes of either B or T cell origin. In keeping with this concept, we were able to demonstrate that in the majority of cases of Hodgkin's disease the interleukin-2 receptor (IL2-R) was detectable on H-RS-cells by three monoclonal antibodies (anti-Tac, Tü69 and ACT-1) with the sensitive alkaline phosphatase anti-alkaline phosphatase (APAAP) tissue staining procedure. In extension of these studies, we precipitated IL2-R antigen from two established permanent cell lines (L540 and L591) derived from patients with Hodgkin's disease. Although these cell lines are different in nature, in that the L540 line shows rearrangement for the T cell receptor beta chain gene and the L591 line displays rearrangement for immunoglobulin genes (gamma heavy and lambda light chain), the antigen precipitated by monoclonal antibodies against IL2-R exhibited a molecular weight of approximately 53 kd on the L591 line and 58 kd on the L540 line. Thus, these data strongly support the view that the labelling of H-RS-cells in tissue sections for IL2-R was not merely due to cross-reactivities of the two anti-IL2-R antibodies with a determinant of an unrelated structure but represents true IL2-R molecules.

IgA Binding Factors and Fc Receptors for IgA: Comparative Studies Between IgA and IgE Fc Receptor Systems

The expression of Fc receptors (FcR) for IgA (Fc alpha R) as well as for IgE (Fc epsilon R) on T lymphocytes (T cells) is enhanced or up regulated by the corresponding class of immunoglobulins (Ig). The production of class-specific regulatory factors binding to IgA and IgE (IgA binding factor [IgA-BF]; IgE binding factor [IgE-BF]) is also induced by these respective ligands. Murine IgA-BFs produced by a T hybridoma T2D4 and concanavalin A-activated spleen cells suppressed the in vitro IgA antibody responses of pokeweed mitogen-stimulated mouse spleen cells class-specifically. Human IgA antibody response was also suppressed by the murine IgA-BF. Similar suppressive IgA-BF is also produced by a human natural killer (NK)-like cell line (YT), which has no rearrangement of the T cell receptor beta-chain gene, indicating that non-T non-B/LGL cells may also be involved in the regulation of the class-specific antibody responses. It appears that, in human as well as murine systems, T- and NK-cells have the capacity to co-express multiple class-specific FcRs and to produce the corresponding immunoglobulin binding factors. While the Fc epsilon R expression is abnormally enhanced in the diseases with hyperimmunoglobulinemia E, disregulation of Fc alpha R is associated with certain human diseases involving the altered IgA regulation. In IgA nephropathy, which is characterized by increased serum IgA level and IgA deposition in the mesangium, there is an enhancement of the expression of Fc alpha R. In contrast, IgA failed to induce Fc alpha R significantly on the lymphocytes from the patients with selective IgA deficiency, indicating that Fc alpha R plays an important role in the IgA regulation in vivo.

Human Acute Unclassified Leukemia With a Unique t(4;17) Chromosomal Translocation Expresses T Lymphoid and Myeloid Surface Antigens After In Vitro Culture

Bilineage differentiation along both the T lymphoid and the myeloid lineage while in in vivo diffusion chamber (DC) and in vitro suspension culture was observed in a case of acute unclassified leukemia (null-AL) and t(4;17). Prior to culture, the blast cells were TdT and la positive but did not express any lineage-specific antigenic markers. Furthermore, the immunoglobulin heavy chain and T cell receptor beta-chain genes were in germline configuration. Cytogenetically, all metaphases had the unique translocation t(4;17) (q25;q23) prior to and after culture, supporting the leukemic origin of the cells. During both DC culture and suspension culture with and without tetradecanoyl-phorbol-acetate (TPA), a substantial increase in the absolute and relative number of cells expressing both myeloid and T lymphoid antigenic markers occurred. Double-fluorescence analysis demonstrated the expression of antigenic markers of both lineages on the same population of cells, and electron microscopy revealed the induction of myeloperoxidase after both DC and suspension culture. Immunoglobulin heavy chain and T cell receptor beta-chain genes remained in germline configuration after treatment with TPA, when analyzed with JH and CT beta probes, respectively. These findings indicate that this case represents a null-AL with dual-lineage capabilities, which has probably arisen from the malignant transformation of a bipotential stem cell of lymphoid and myeloid progeny.

7q32-q36 Translocations in Childhood T Cell Leukemia: Cytogenetic Evidence for Involvement of the T Cell Receptor /?-Chain Gene

Blast cell chromosomal rearrangements involving the long arm of chromosome 7 were identified in eight of 197 cases of childhood acute lymphoblastic leukemia (ALL). Breakpoints were variable but tended to cluster in either the proximal or the terminal 7q region, depending on the immunophenotype of the cells. The 7q32-q36 region, the locus of the T cell receptor beta-chain gene, was the site of breakpoints in four of 31 cases of T cell ALL but was not involved in any of the 166 cases originating from B cell precursors (P less than .0004). In three of the four T cell cases it was possible to identify the chromosomal segment that had been translocated to the 7q32-q36 region: 1p32, 2p21, and 6p21. The 1p32 and 6p21 bands are particularly interesting, as they contain the sites of two known protooncogenes, c-L-myc and hpim, respectively. Our findings suggest that the locus of the beta-chain gene of the T cell receptor is a preferential site for certain chromosomal rearrangements in leukemic T lymphoblasts, analogous to the T cell receptor alpha-chain gene on human chromosome 14. Translocation of proto-oncogenes to a site near the beta-chain regulatory sequences provides a potential mechanism for oncogene activation.

T cell receptor gamma chain variable gene rearrangements in acute lymphoblastic leukemias of T and B lineage.

The status of immunoglobulin and T cell receptor genes in acute lymphoblastic leukemia (ALL) of T and B lineage has been studied. Our data indicate that illegitimate gene rearrangements at immunoglobulin heavy chain (in T cell ALL), and T cell receptor beta chain (in pre-B ALL) genes are only rarely found (2 out of 30 patients). In contrast, T cell receptor gamma chain gene rearrangements, characteristically found in T-ALL, are also present in 7 of 18 patients with pre-B ALL. Several features distinguish these illegitimate T cell receptor gamma chain gene rearrangements from those in normal and leukemic T cells. V gamma genes located far upstream of the J gamma/C gamma complexes (V gamma 2, V gamma 3, V gamma 4, V gamma 5) appear to be preferentially used in normal adult peripheral blood T cells. In contrast, V gamma genes located immediately 5' to the J gamma/C gamma complexes (V gamma 8, V gamma 9, V gamma 10, V gamma 11) predominate in V gamma -J gamma recombinations observed in T-ALL and pre-B ALL. Whereas the J gamma 2 region is primarily used in T cell receptor gamma gene rearrangements observed in T-ALL, those in pre-B ALL are confined mostly to the J gamma 1 region. These data suggest a limited accessibility of the T cell receptor gamma chain gene locus for recombination processes in early stages of B cell differentiation.

T Cell Receptor Beta Chain and Immunoglobulim Heavy Chain Switch Region Gene Polymorphisms Associated with Membranous Nephropathy

T Cell Receptor Beta Chain and Immunoglobulim Heavy Chain Switch Region Gene Polymorphisms Associated with Membranous Nephropathy

Correlation of T cell receptor gene rearrangements to T cell surface antigen expression and to serum immunoglobulin level in scid mice.

The severe combined immunodeficient (scid) mouse which has undetectable serum immunoglobulin (Ig) contains a small number of thymic lymphocytes which express Thy-1 and IL2 receptors (IL2R) but not Lyt-2 or L3T4 molecules. These thymocytes did not show any rearrangement of T cell receptor (TCR) beta-chain genes. Such thymocyte characteristics in the scid mouse were similar to the 15-day embryonic thymocytes in ordinary mice, indicating that the scid mouse thymocytes are arrested in the early stage of intrathymic differentiation. However, low or medium level serum Ig was occasionally found in the littermates of the scid mouse. The thymocytes of these mice showed some evidence of TCR beta-chain gene rearrangement and the presence of Lyt-2+/L3T4+ cells in correlation with the serum Ig level. In the mice with some serum Ig the thymocyte cell number was increased and the proportion of IL2R+ cells was decreased. Collectively, these results suggest that the rearrangement of TCR beta-chain genes is associated with the expression of Lyt-2 and L3T4 molecules in intrathymic differentiation and probably with cell proliferation of the migrated lymphoid cells in the scid mouse.

Human acute unclassified leukemia with a unique t(4;17) chromosomal translocation expresses T lymphoid and myeloid surface antigens after in vitro culture

Bilineage differentiation along both the T lymphoid and the myeloid lineage while in in vivo diffusion chamber (DC) and in vitro suspension culture was observed in a case of acute unclassified leukemia (null-AL) and t(4;17). Prior to culture, the blast cells were TdT and la positive but did not express any lineage-specific antigenic markers. Furthermore, the immunoglobulin heavy chain and T cell receptor beta- chain genes were in germline configuration. Cytogenetically, all metaphases had the unique translocation t(4;17) (q25;q23) prior to and after culture, supporting the leukemic origin of the cells. During both DC culture and suspension culture with and without tetradecanoyl- phorbol-acetate (TPA), a substantial increase in the absolute and relative number of cells expressing both myeloid and T lymphoid antigenic markers occurred. Double-fluorescence analysis demonstrated the expression of antigenic markers of both lineages on the same population of cells, and electron microscopy revealed the induction of myeloperoxidase after both DC and suspension culture. Immunoglobulin heavy chain and T cell receptor beta-chain genes remained in germline configuration after treatment with TPA, when analyzed with JH and CT beta probes, respectively. These findings indicate that this case represents a null-AL with dual-lineage capabilities, which has probably arisen from the malignant transformation of a bipotential stem cell of lymphoid and myeloid progeny.

CS-A therapy in MRL-lpr/lpr mice: amelioration of immunopathology despite autoantibody production.

MRL-lpr/lpr mice spontaneously develop massive T cell lymphadenopathy, autoantibodies, and immune-mediated pathology. These mice are thought to be models of various human autoimmune diseases, including systemic lupus, Sjogren's syndrome, and rheumatoid arthritis. We have used cyclosporin A (CS-A) treatment as a tool by which the mechanisms of immune-mediated pathology might be dissected. CS-A was used because of its known preferential inhibition of T cell function and the marked expansion in MRL-lpr/lpr mice of an unusual L3T4-, Lyt-2-, 6B2+ T cell population. CS-A prevented lymphadenopathy and expansion of L3T4-, Lyt-2-, 6B2+ T cells in the peripheral lymph nodes, and also in the thymus. The increased expression of the c-myb and T cell receptor beta-chain genes associated with these unusual cells was also corrected. The finding of increased numbers of L3T4-, Lyt-2-, 6B2+ thymocytes in untreated mice suggests abnormal intrathymic differentiation in lpr/lpr mice, a defect that was corrected by CS-A. Treated mice had a marked decrease in arthritis and glomerulonephritis and significantly prolonged survival. These beneficial effects of CS-A occurred despite a lack of reduction in antibodies reactive with DNA, circulating immune complexes, rheumatoid factor titers, or immunoglobulin concentrations. These results demonstrate that the B cell hyperactivity of MRL-lpr/lpr mice can proceed without the T cell proliferative disease.

Adult-onset cyclic neutropenia is a benign neoplasm associated with clonal proliferation of large granular lymphocytes.

Human cyclic neutropenia occurs in children and adults. Adult-onset cyclic neutropenia is an acquired disease characterized by increased numbers of large granular lymphocytes (LGL), in contrast to childhood-onset cyclic neutropenia in which LGL counts are normal. We investigated the clonality of lymphocytes in these two groups of patients by assessing the rearrangement status of the T cell receptor beta chain gene. Patients with adult-onset cyclic neutropenia showed clonal rearrangement of the T beta gene whereas the children did not. Since LGL are known to have multiple regulatory effects on normal hematopoiesis, the finding of a clonal proliferation of this lymphocyte population implicates these cells in the pathogenesis of cyclic neutropenia.

Kappa-chain gene rearrangement in an apparent T-lineage lymphoma.

We describe a 10-yr-old boy with T-lineage non-Hodgkin's lymphoma. He had a mediastinal mass, swollen supraclavicular lymph nodes, and pleural effusion. A supraclavicular lymph node biopsy under light microscopy showed a malignant lymphoma of diffuse lymphoblastic type. Most of the cells taken from the malignant pleural effusion expressed T cell-associated antigens such as Leu-1 and OKT 8. To confirm these antigens as T-lineage lymphoma, we examined genomic DNA from malignant cells obtained from the pleural effusion. As was expected, T cell receptor beta-chain gene rearrangements were demonstrated. However, when the immunoglobulin gene organization was analyzed, we detected rearrangements in both the heavy- and kappa-chain genes. To our knowledge, this is the first case in which kappa-chain gene rearrangement was detected in apparent T-lineage cells. These findings provide important information relating to determination of the cellular lineage of lymphoid malignancy.

Morphology and lytic mechanisms of interleukin 3-dependent natural cytotoxic cells: tumor necrosis factor as a possible mediator.

Populations of interleukin 3 (IL 3)-dependent cells can be derived from mouse bone marrow that display natural cytotoxicity (NC) against Wehi-164 target cells but do not display natural killing against YAC-1 cells. These bone marrow-derived NC cells cultured up to 2 mo in IL 3 do not contain rearranged T cell receptor beta-chain genes. They appear to be mast-like cells by electron microscopy and contain heterogeneous type granules. The molecules that mediate NC appear to be contained in these granules and are preformed because protein synthesis inhibitors have no effect on the capacity of IL 3-dependent NC cells to lyse Wehi-164 target cells. In addition to the IL 3-dependent bone marrow-derived cells, the basophilic leukemia cells, RBL-1, but not P815 mastocytoma cells were found to mediate NC against Wehi-164 cells. Both bone marrow-derived NC and RBL-1 cells can lyse L929 cells in 18 hr, suggesting that the putative NC mediator may be related to lymphotoxin/tumor necrosis factor (TNF). Recombinant human TNF displayed identical properties as NC cells; both entities possessed the same target cell specificity and had similar kinetics of target cell killing. The use of polyclonal rabbit antimouse TNF antibody blocked the actions of NC cells. Thus we believe that the mediation of NC is through the actions of a TNF-like molecule.

Cutaneous pleomorphic T cell lymphoma: Immunologic, virologic, and T-cell receptor gene rearrangement studies in one European case with initial pseudolymphoma presentation

An unusual case of cutaneous nodular T cell lymphoma evolving for 4 years with massive eosinophilia and greatly increased IgE levels is discussed. Repeated histologic and immunohistologic examinations could not ascertain malignancy because tumors were composed of a granuloma-like, highly polymorphic cellular infiltrate with mature immunotype and no significant nuclear abnormalities nor epidermotropism. T cell lymphoma was evidenced by the T cell receptor beta-chain gene study, which showed a clonal rearrangement. Final histologic classification was "pleomorphic T cell lymphoma" because further biopsy samples displayed numerous lymphoid cells with pleomorphic convoluted nuclei. The T-cell receptor gene probe is a major tool for the early diagnosis of some T cell lymphomas. The case we report shares many features with the smoldering type of human T cell lymphotropic virus type I-associated Japanese endemic pleomorphic T cell lymphoma. Our virologic study confirms that in nonendemic Western countries, pleomorphic T cell lymphomas do not show evidence of a retrovirus association.