RESUMEN
To characterize the T cell receptor (TCR) repertoire expressed by the V delta 1+ gamma/delta T cell population, we have studied the V delta 1-J delta 1 junctional sequences from peripheral blood samples of healthy donors. We show that, surprisingly, this repertoire is restricted in most healthy adults, with a donor-specific and relatively stable pattern, whereas this repertoire remains unrestricted in infants, and is similar to that of thymocytes. These data contrast with the general assumption that the junctional repertoire of V delta 1+ gamma/delta T cells is extensive, and strongly suggest that peripheral recruitment of V delta 1+ cells bearing particular TCR occurs in humans during the postnatal stage.
Asunto(s)
Receptores de Antígenos de Linfocitos T gamma-delta/genética , Linfocitos T/inmunología , Adulto , Envejecimiento/inmunología , Secuencia de Bases , Preescolar , Reordenamiento Génico de la Cadena delta de los Receptores de Antígenos de los Linfocitos T , Humanos , Lactante , Datos de Secuencia Molecular , Receptores de Antígenos de Linfocitos T gamma-delta/análisisRESUMEN
T cell rearranging gene gamma (TRG gamma) and T cell antigen receptor beta (TCR beta) chain gene rearrangement and transcription were studied in a series of patients with B-lineage acute lymphoblastic leukemia (ALL), in which the Ig H chain genes are rearranged and the surface phenotype reproduces the stages of normal pre-B maturation. For comparison, polyclonal T cells from peripheral blood of healthy donors and blast cells from 19 cases of T lineage ALL were also studied. In this study we demonstrate the presence of a clonal rearrangement of the TRG gamma in 18 of the 22 B-lineage ALL cases and establish that this rearrangement, which generally involves the J gamma 1 region, is often monoallelic and appears different from the biallelic J gamma 2 rearrangement frequently seen in T-cell ALLs. In 9 of 22 cases, we found rearrangement of the genes of the TCR beta chain, which never involved the J beta 1 region. Conversely, the TRG gamma were seen in germline configuration in all 19 cases of B chronic lymphoid malignancies. In none of the 9 AML cases studied was TRG gamma and TCR beta chain gene rearrangement found. The TCR beta chain genes were rearranged in one B cell chronic lymphocytic leukemia (CLL). We also show that in B-lineage ALL, the cells probably use the same V gamma genes for TRG gamma rearrangements as the malignant cells in T-ALL and the polyclonal T cells. In none of the 13 B-lineage ALL cases investigated by Northern analysis was TCR beta mRNA expression detected, whereas a weak expression of TRG gamma transcripts was found in two of these cases. The correlations between surface phenotype, rearrangement of TRG gamma, TCR beta, and Ig H chain genes were analyzed. The significance of rearrangement of TRG gamma and TCR beta chain genes in B or pre-B cells is also discussed.
Asunto(s)
Linfocitos B/análisis , Leucemia de Células Pilosas/genética , Leucemia Linfoide/genética , Fragmentos de Péptidos/genética , Receptores de Antígenos de Linfocitos T/genética , Genes , Humanos , Cadenas Pesadas de Inmunoglobulina/genética , Leucemia de Células Pilosas/inmunología , Leucemia Linfoide/inmunología , ARN Mensajero/análisis , ARN Neoplásico/análisis , Receptores de Antígenos de Linfocitos T alfa-beta , Receptores de Antígenos de Linfocitos T gamma-delta , Linfocitos T/análisisRESUMEN
We have defined transcriptional enhancing sequences inside the TCR-delta gene locus, using transient transfections with constructs containing DNA fragments cloned upstream to a reporter gene fused to a heterologous promoter. A 14-kb DNA region extending from the J delta 3 segment to 6 kb 3' to C delta was analyzed. We show the presence of positive regulatory sequences inside the J delta 3-C delta intron and have localized these sequences to two DNA fragments of approximately 300 and 258 bp. Analysis of cell specificity of the activation of such sequences demonstrates a T cell pattern for one of the two fragments. The nucleotide sequence of the T cell-specific element shows motifs sharing homology with previously described core enhancers.
Asunto(s)
Elementos de Facilitación Genéticos , Regulación de la Expresión Génica , Genes , Intrones , Receptores de Antígenos de Linfocitos T/genética , Linfocitos T/inmunología , Secuencia de Bases , Clonación Molecular , Humanos , Sustancias Macromoleculares , Datos de Secuencia Molecular , Mapeo RestrictivoRESUMEN
Southern blot analysis using a genomic probe of the human TCR-gamma chain first variable gene subgroup (V gamma I) was performed on DNA samples from both parents of 36 healthy Caucasian families. Two types of polymorphisms were found in these 72 unrelated DNA samples: three repertoire polymorphisms and two restriction fragment length polymorphisms (RFLP). In all cases, Mendelian inheritance of these polymorphisms was demonstrated. The most frequent repertoire polymorphism consists in the lack of the V gamma 4 and V gamma 5 segments. In 16% of chromosomes, the Eco RI and Taq I restriction fragments corresponding to V gamma 4 and V gamma 5 were lacking, with no additional bands. In these cases, a decrease of 10 kb was observed in the Bam HI fragment containing all V gamma I segments as compared with samples containing V gamma 4-V gamma 5 segments. To better understand this polymorphism, which takes place in a previously incompletely defined region, the central part of the V gamma I region, including the polymorphic V gamma 4-V gamma 5 segments, was cloned. This allowed us to localize precisely the V gamma 5 segment and thus complete the description of the V gamma I region. A striking homology of DNA and deduced amino acid sequences is present between V gamma 2 and V gamma 4 and between V gamma 3 and V gamma 5, much higher than that observed between V gamma 2 and V gamma 3 and between V gamma 4 and V gamma 5. The differences in nucleotide sequence occur mainly in the intron and three hypervariable regions. These results strongly suggest a gene duplication relationship between the segments V gamma 2-V gamma 3 and the segments V gamma 4-V gamma 5. The most frequent RFLP documented in this study is due to the combined absence of the Eco RI and the Taq I sites located in the noncoding region between V gamma 3 and V gamma 4. The haplotypic frequence of this RFLP is 6.9% of the general population. As the gamma/delta receptor may play an important role in immunological response, the biological relevance of the high degree of polymorphism occurring in the V gamma I region, as well as its possible association with some immune disturbances, should be further explored.
Asunto(s)
ADN/genética , Polimorfismo Genético , Receptores de Antígenos de Linfocitos T/genética , Secuencia de Aminoácidos , Secuencia de Bases , Southern Blotting , Línea Celular Transformada , Clonación Molecular , Sondas de ADN , Desoxirribonucleasa BamHI , Desoxirribonucleasa EcoRI , Desoxirribonucleasas de Localización Especificada Tipo II , Humanos , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Polimorfismo de Longitud del Fragmento de Restricción , Homología de Secuencia de Ácido NucleicoRESUMEN
Recently, we and others described a new chromosomal rearrangement, that is, inv(7)(p15q34) and t(7;7)(p15;q34) involving the T-cell receptor beta (TCRbeta) (7q34) and the HOXA gene locus (7p15) in 5% of T-cell acute lymphoblastic leukemia (T-ALL) patients leading to transcriptional activation of especially HOXA10. To further address the clinical, immunophenotypical and molecular genetic findings of this chromosomal aberration, we studied 330 additional T-ALLs. This revealed TCRbeta-HOXA rearrangements in five additional patients, which brings the total to 14 cases in 424 patients (3.3%). Real-time quantitative PCR analysis for HOXA10 gene expression was performed in 170 T-ALL patients and detected HOXA10 overexpression in 25.2% of cases including all the cases with a TCRbeta-HOXA rearrangement (8.2%). In contrast, expression of the short HOXA10 transcript, HOXA10b, was almost exclusively found in the TCRbeta-HOXA rearranged cases, suggesting a specific role for the HOXA10b short transcript in TCRbeta-HOXA-mediated oncogenesis. Other molecular and/or cytogenetic aberrations frequently found in subtypes of T-ALL (SIL-TAL1, CALM-AF10, HOX11, HOX11L2) were not detected in the TCRbeta-HOXA rearranged cases except for deletion 9p21 and NOTCH1 activating mutations, which were present in 64 and 67%, respectively. In conclusion, this study defines TCRbeta-HOXA rearranged T-ALLs as a distinct cytogenetic subgroup by clinical, immunophenotypical and molecular genetic characteristics.
Asunto(s)
Proteínas de Homeodominio/genética , Leucemia-Linfoma de Células T del Adulto/genética , Receptores de Antígenos de Linfocitos T alfa-beta/genética , Adolescente , Adulto , Niño , Deleción Cromosómica , Inversión Cromosómica , Femenino , Reordenamiento Génico de Linfocito T , Proteínas Homeobox A10 , Humanos , Inmunofenotipificación , Leucemia-Linfoma de Células T del Adulto/patología , Leucemia-Linfoma de Células T del Adulto/fisiopatología , Masculino , Persona de Mediana Edad , Receptor Notch1/genética , Activación Transcripcional , Translocación GenéticaRESUMEN
To provide a sensitive and generally applicable method to detect clonal cells in acute lymphoblastic leukemias (ALL), we have designed a new strategy based on the polymerase chain reaction (PCR) amplification of the T cell receptor gamma delta gene rearrangements found in most T and B lineage ALLs. PCR allows rapid sequencing of variable-(diversity)-joining (V-[D]-J) junctions from tumor DNA and construction of anti-junctional oligonucleotides (AJOs) used as probes to detect clonal cells in the same patient. We have defined oligonucleotides suitable for all T cell receptor (TCR) rearrangements involving functional V gamma segments. Oligonucleotides corresponding to preferential TCR delta rearrangements in T and B lineage ALLs were also used. By analysis of the nucleotide sequence of 52 V gamma-V gamma junctions from 30 cases of B and T ALLs, we demonstrate that V-J junctional sequences are clone specific in both lineages and at all stages of differentiation examined despite the frequent presence of the recently described P nucleotides. Experiments performed with TCR gamma delta AJOs on DNA from tumor cells and polyclonal T cells show that AJOs can be used to differentiate clonal cells from polyclonal T cells, distinguish between different T cell clones, and detect residual clonal populations at 10(-4)/10(-5) dilution. AJOs were also used to detect residual disease in samples from patients in clinical and morphological complete remission. Finally, rearrangement patterns were studied by classical Southern analysis in selected cases at both presentation and subsequent relapse showing absence of clonal evolution in most cases. V-(D)-J nucleotide sequences of rearrangements with an identical pattern of rearrangement at presentation and relapse were identical in all cases analyzed. We therefore describe a new, specific, and clinically useful strategy for the detection of minor clonal populations applicable in the majority of cases of ALL.
Asunto(s)
Reordenamiento Génico de la Cadena gamma de los Receptores de Antígenos de los Linfocitos T , Sondas de Oligonucleótidos , Leucemia-Linfoma Linfoblástico de Células Precursoras/diagnóstico , Receptores de Antígenos de Linfocitos T/genética , Alelos , Secuencia de Bases , Células Clonales , Humanos , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/patología , Receptores de Antígenos de Linfocitos T gamma-deltaRESUMEN
Strong expression of at least one of the three D-type cyclins is common in human cancers. While the cyclin D1 and D3 genes (CCND1 and CCND3) are recurrently involved in genomic rearrangements, especially in B-cell lymphoid neoplasias, no clear involvement of the cyclin D2 gene (CCND2) has been reported to date. Here, we identified chromosomal translocations targeting the CCND2 locus at 12p13, and the T-cell receptor beta (TCRB) or the TCRA/D loci in T-cell acute lymphoblastic leukemias (T-ALLs). Expression analysis demonstrated dramatic cyclin D2 overexpression in the translocated cases (n=3) compared to other T-ALLs (total, n=89). In order to evaluate dysregulation in T-ALL with respect to normal T-cell differentiation, we analyzed CCND2 expression in normal purified human thymic subpopulations. CCND2 levels were downregulated through progression from the early stages of human T-cell differentiation, further suggesting that the massive and sustained expression in the CCND2-rearranged T-ALL cases was oncogenic. Association with other oncogene expression (TAL1, HOXAs, or TLX3/HOX11L2), NOTCH1 activating mutations, and/or CDKN2A/p16/ARF deletion, showed that cyclin D2 dysregulation could contribute to multi-event oncogenesis in various T-ALL groups. This report is the first clear evidence of a direct involvement of cyclin D2 in human cancer due to recurrent somatic genetic alterations.
Asunto(s)
Cromosomas Humanos Par 12/genética , Ciclinas/biosíntesis , Ciclinas/genética , Leucemia-Linfoma de Células T del Adulto/genética , Receptores de Antígenos de Linfocitos T/genética , Translocación Genética , Adulto , Separación Celular , Niño , Ciclina D2 , Análisis Citogenético , Análisis Mutacional de ADN , Reordenamiento Génico , Humanos , Leucemia-Linfoma de Células T del Adulto/diagnóstico , Leucemia-Linfoma de Células T del Adulto/metabolismoRESUMEN
Seventeen patients with adult T-cell leukemia (ATL) and 21 with tropical spastic paraparesis/human T-cell leukemia/lymphoma virus type I (HTLV-I)-associated myelopathy (TSP/HAM) were observed during a 3-yr survey (1986-1988) in some hospitals in Paris, France. Most of them were black, originating from high-HTLV-I-endemic areas (West Indies or Africa), but two cases of TSP/HAM occurred in French Caucasians. In one case, the patient acquired the virus from a transfusion during a cardiac transplantation. Most of the ATL cases were diagnosed as acute leukemia or lymphoma, with a proliferation of CD2+, CD3+, CD4+, CD8-, DR+, and CD25+ lymphoid cells. Only three cases were diagnosed as a smoldering ATL. All of the TSP/HAM cases exhibited a spastic paraparesis with a chronic and slow evolution and high HTLV-I antibody titers in serum and cerebrospinal fluid, with a high HTLV-I antibody index and specific HTLV-I immunoglobulin = oligoclonal bands. In TSP/HAM, a high percentage of DR-expressing cells (15 to 40%) was found, with a slightly elevated CD4/CD8 ratio. This was associated with the presence of 1 to 10% abnormally shaped nuclei in lymphoid cells and a polyclonal integration of HTLV-I proviruses in these peripheral blood mononuclear cells. On the contrary, a clonal integration was always found in the ATL malignant cells (leukemic, lymph node, and cutaneous infiltrate). Long-term interleukin 2-dependent T-cell lines (CD2+, CD3+, CD4+, and WT31+) with activated T-cell markers (CD25+ and DR+) producing HTLV-I were established from ATL and TSP/HAM peripheral blood mononuclear cells.
Asunto(s)
Leucemia-Linfoma de Células T del Adulto/epidemiología , Paraparesia Espástica Tropical/epidemiología , Antígenos CD/análisis , Femenino , Francia/epidemiología , Productos del Gen env/análisis , Anticuerpos Anti-HTLV-I/análisis , Humanos , Leucemia-Linfoma de Células T del Adulto/inmunología , Masculino , Paraparesia Espástica Tropical/inmunologíaRESUMEN
The t(X;14)(q28;q11.2) translocation is associated with mature T-cell proliferations. Recently this translocation has been shown to implicate the MTCP-1/c6.1B gene on chromosome Xq28, leading to aberrant or overexpressed MTCP-1 transcripts. The potential coding role of this gene was made uncertain by the lack of a long open reading frame in its major transcripts. However, a short 204 bases open reading frame is potentially coding for a 68 amino-acid protein. Here, we show that this open reading frame sequence and the deduced product are well conserved in mouse. A 8 kD protein (p8), which corresponds to the predicted molecular weight was revealed in transient transfectants and in cell lines by Western blotting, using a rabbit antiserum. This product was absent in lymphoblastoid cell lines with deletions of the MTCP-1/c6.1B locus. A dramatic overexpression of p8 was found in leukemic cells from a patient with a t(X;14). This small protein was localized in the cytoplasm by immunofluorescence. In conclusion, MTCP-1 encodes for a cytoplasmic 8 kD product. Its potential role in leukemogenesis is supported by its overexpression in leukemia with t(X;14), but its function remains unknown.
Asunto(s)
Cromosomas Humanos Par 14 , Leucemia de Células T/genética , Proteínas de Neoplasias/genética , Translocación Genética , Cromosoma X , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Citoplasma/metabolismo , ADN Complementario , Humanos , Sueros Inmunes , Leucemia de Células T/metabolismo , Ratones , Datos de Secuencia Molecular , Oncogenes , Proteínas Proto-Oncogénicas/genética , Proteínas Proto-Oncogénicas/inmunología , Células Tumorales CultivadasRESUMEN
T-cell lymphoproliferative diseases are often associated with recurrent chromosomal translocations involving T cell receptor genes (TCR) and genes that are thought to play a role in the pathogenesis of these diseases. Whereas numerous such genes have already been identified in acute T cell leukemias, no candidate gene has yet been identified to play a role in the heterogeneous group of T cell proliferations with a mature phenotype. We here report the molecular cloning of two examples of the rare but recurrent t(X;14) translocation. The first translocation was associated with a benign clonal proliferation in an ataxia telangiectasia patient and the second with a T cell prolymphocytic leukemia. Both translocations implicated the TCR alpha/delta locus and a common breakpoint region on chromosome Xq28. A previously unidentified gene, abnormally transcribed in both T cell proliferations, was characterized in the immediate proximity of the breakpoints. This Xq28 gene has no homology with known sequences, uses a complex alternative splicing pattern and demonstrates two short open reading frames. This gene, named MTCP-1 (Mature T Cell Proliferation-1) is the first candidate gene potentially involved in the leukemogenic process of mature T cell proliferations.
Asunto(s)
Trastornos Linfoproliferativos/genética , Proteínas Proto-Oncogénicas/genética , Receptores de Antígenos de Linfocitos T alfa-beta/genética , Translocación Genética , Cromosoma X , Empalme Alternativo , Secuencia de Aminoácidos , Ataxia Telangiectasia/genética , Secuencia de Bases , Cromosomas Humanos Par 14 , Expresión Génica , Genes , Humanos , Leucemia Prolinfocítica/genética , Datos de Secuencia Molecular , ARN Mensajero/genética , Mapeo Restrictivo , Linfocitos T/citologíaRESUMEN
The MTCP1 gene is involved in the t(X;14)(q28;q11) translocation associated with T-cell prolymphocytic leukemia and related conditions. This gene is unusual in that it codes for two distinct proteins: a small mitochondrial protein, p8MTCP1, and a putative oncogenic protein, p13MTCP1. Scarcity of material from t(X;14)-associated proliferations and very low levels of mRNA expression have so far prevented a thorough description of p13MTCP1-encoding transcripts. Here, we characterize two additional t(X;14) bearing leukemias allowing this analysis. In one case, with a breakpoint located 5' to the MTCP1 gene, the level of transcription of previously described p13MTCP1-encoding transcripts is enhanced. In the second case, with a breakpoint within the MTCP1 intron I, an alternative transcription initiation site is demonstrated in the tumor cells at 229 bp upstream to exon II. The identification of this internal promoter, together with the similarity between TCL1 and MTCP1 genomic structures, allow us to propose a model in which the duplication of an ancestral gene was followed by the insertion of one copy within the intron of a p8-encoding gene, accounting for the unusual feature of the MTCP1 gene.
Asunto(s)
Leucemia de Células T/genética , Transcripción Genética , Translocación Genética , Anciano , Secuencia de Bases , División Celular/genética , Femenino , Humanos , Leucemia Prolinfocítica/genética , Persona de Mediana Edad , Datos de Secuencia Molecular , Proteínas Proto-Oncogénicas/genética , Linfocitos T/fisiologíaRESUMEN
An unusually small (8 kD) protein (p8MTCP-1) is coded by the putative oncogene MTCP-1 (also called c6.1B), involved in the translocation t(X;14)(q28;q11) associated with some mature T-cell proliferations. Here, we show by subcellular fractionation and by confocal microscopy that this protein is located in the mitochondria. This localization orientates toward a role of p8MTCP-1 in the mitochondrial metabolism which may be relevant for the oncogenic process.
Asunto(s)
Mitocondrias/metabolismo , Secuencia de Aminoácidos , Técnica del Anticuerpo Fluorescente , Humanos , Microscopía Confocal , Datos de Secuencia Molecular , Proteínas Proto-Oncogénicas/química , Proteínas Proto-Oncogénicas/genética , Homología de Secuencia de AminoácidoRESUMEN
PURPOSE: Most studies using various reverse-transcription polymerase chain reaction (RT-PCR) techniques reported that the detection of the AML1-ETO fusion transcript was a common finding in long-term complete remission (CR) in acute myeloid leukemia (AML) with t(8;21) translocation. However, larger prospective studies with interlaboratory quality control may be important to investigate more precisely the clinical usefulness of studying minimal residual disease with RT-PCR in t(8;21) AML. PATIENTS AND METHODS: We collected 223 marrow samples from 51 patients with t(8;21) AML diagnosed in five centers and tested all samples by two different RT-PCR techniques (a nested technique and a one-step technique, with a sensitivity of 10(-6) and 10(-5), respectively) in two different laboratories. RESULTS: Samples from 14 patients in long persistent CR (median follow-up duration, 112 months) were taken at least twice, and all were PCR-negative by both techniques. Samples were prospectively taken from 37 patients after achievement of first CR and/or second CR, before intensive consolidation treatment, and every 3 to 6 months after completion of therapy. Patients who converted to PCR negativity with the one-step technique (60%) or both techniques (48%) after CR achievement had a longer CR duration than those with persistently positive PCR results (two-sided log-rank test, P =.0001). Patients who became PCR-negative with the one-step technique before intensive consolidation (23%) had a lower relapse rate (11% v 72%) and a longer CR duration than those who remained persistently PCR-positive at that point (two-sided log-rank test, P =.0015). CONCLUSION: Patients with AML with t(8;21) in long-term remission were all PCR-negative. In prospectively studied patients, a good correlation was found between negative PCR results and absence of relapse. Early negative results with the one-step RT-PCR technique, before consolidation treatment, seemed to carry an especially good prognosis, suggesting that RT-PCR analysis could help in choosing the type of consolidation therapy in patients with t(8;21) AML.
Asunto(s)
Reordenamiento Génico , Leucemia Mieloide/genética , Reacción en Cadena de la Polimerasa , Transcripción Genética , Enfermedad Aguda , Adolescente , Adulto , Fusión Artificial Génica , Células de la Médula Ósea/patología , Niño , Preescolar , Subunidad alfa 2 del Factor de Unión al Sitio Principal , Estudios de Evaluación como Asunto , Femenino , Estudios de Seguimiento , Humanos , Leucemia Mieloide/terapia , Modelos Lineales , Masculino , Persona de Mediana Edad , Neoplasia Residual , Proteínas de Fusión Oncogénica/genética , Pronóstico , Estudios Prospectivos , Proteína 1 Compañera de Translocación de RUNX1 , Inducción de Remisión , Estudios Retrospectivos , Sensibilidad y Especificidad , Factores de Transcripción/genética , Translocación Genética/genéticaRESUMEN
Acute lymphoblastic leukemia (ALL) is the most common cancer in children. In recent years, the outcome has been globally improved by current therapies, but it remains poor in patients with high, persistent residual disease following the first course of chemotherapy, prompting evaluation of the possible beneficial effects of immunotherapy protocols. In this study, we hypothesized that the disruption of two immunoregulatory pathways controlling the auto-reactive T cell response might synergize with dendritic cell-based immunotherapy of the disease, which is considered to be poorly immunogenic. In this study, we used TAL1xLMO1 leukemia cells adoptively transferred in mice, to generate murine leukemia with poorly immunogenic cells as a model for human T-ALL. Subsequently, these animals were treated with several different immunotherapeutic protocols. We compared the efficiency of a classical, dendritic cell-based immunotherapy (injection of dendritic cells loaded with tumor-derived antigenic products), to a combined treatment associating injection of antigen-loaded dendritic cells and disruption of the two immunoregulatory pathways: CD25+ suppressive T cells and cytotoxic T lymphocyte-associated antigens (CTLA-4). We show that this combined treatment resulted in cure, concomitantly with in vivo generation of immune memory, and TNF-alpha secretion. This study demonstrates that the disruption of these two immunoregulatory pathways synergized with immunostimulation by dendritic cells loaded with tumor-derived antigens, and paves the way for the testing of this combination in clinical trials.
Asunto(s)
Inmunoterapia Adoptiva , Leucemia-Linfoma Linfoblástico de Células T Precursoras/inmunología , Leucemia-Linfoma Linfoblástico de Células T Precursoras/terapia , Transducción de Señal/inmunología , Linfocitos T/inmunología , Animales , Antígenos CD/metabolismo , Antígenos de Neoplasias/administración & dosificación , Antígeno CTLA-4 , Línea Celular Tumoral , Células Dendríticas/inmunología , Células Dendríticas/metabolismo , Células Dendríticas/trasplante , Modelos Animales de Enfermedad , Memoria Inmunológica , Subunidad alfa del Receptor de Interleucina-2/biosíntesis , Depleción Linfocítica , Ratones , Ratones Endogámicos C3H , Ratones Endogámicos C57BL , Leucemia-Linfoma Linfoblástico de Células T Precursoras/metabolismo , Linfocitos T/metabolismo , Linfocitos T Reguladores/inmunología , Linfocitos T Reguladores/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Fifty-three hairy cell leukemia patients were treated with low dose recombinant and natural alpha-interferon for 7 or 13 months with a marked improvement in peripheral blood and bone marrow findings. Our results suggest that daily injections have no advantage over three weekly injections, but longer treatment (12-13 months) is better than shorter treatment (6-7 months). The possibility of a relapse raises the issue of the place of splenectomy in the treatment strategy. Since the definition of remission and of relapse is based on the histological changes in the bone marrow, the necessity for the quantification of the hairy cell infiltration on bone marrow sections is stressed.
Asunto(s)
Interferón Tipo I/uso terapéutico , Leucemia de Células Pilosas/terapia , Relación Dosis-Respuesta a Droga , Tolerancia a Medicamentos , Hematopoyesis , Humanos , Interferón Tipo I/efectos adversos , Recuento de Leucocitos , Recuento de Plaquetas , Proteínas Recombinantes/uso terapéutico , Esplenectomía , Factores de TiempoRESUMEN
A case of clinically typical CML (300 x 10(6)/l leukocytes, 400 x 10(6)/l platelets, splenomegaly) is presented. After complete remission induced by busulphan, no clinical or haematological abnormalities were observed for 27 years until the development of acute leukaemia (type M1), which was rapidly fatal after a brief chemotherapy-induced remission. The cytogenetic findings were also original: no chromosome Ph1 (during remission 3 years after the onset of the disease), no translocation (banding study 5 years later), and no bcr/abl rearrangement (during the terminal phase).
Asunto(s)
Leucemia Mielógena Crónica BCR-ABL Positiva/patología , Leucemia/patología , Busulfano/uso terapéutico , Femenino , Humanos , Leucemia/genética , Leucemia Mielógena Crónica BCR-ABL Positiva/tratamiento farmacológico , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Persona de Mediana Edad , Factores de TiempoRESUMEN
In order to improve the level of sensitivity and specificity of detection of bone marrow minimal residual disease in the acute lymphoid leukemias, we have performed amplification by the polymerase chain reaction of T cell receptor gamma chain gene rearrangements. Cloning and sequencing of amplified leukemic DNA allowed the construction of a clone-specific anti-junctional oligonucleotide to be used for subsequent detection of minimal infiltration by this clone. Using such an oligonucleotide, it was possible to distinguish clonal DNA from polyclonal T lymphocytes and to detect infiltration by this clone at 10(-6) dilution into germline DNA. We therefore describe a generally applicable method for the detection of minimal residual disease in both T-ALL and the majority of B lineage ALLs.
Asunto(s)
Reordenamiento Génico de la Cadena gamma de los Receptores de Antígenos de los Linfocitos T , Leucemia-Linfoma Linfoblástico de Células Precursoras/diagnóstico , Receptores de Antígenos de Linfocitos T/genética , Linfocitos T/patología , Amplificación de Genes , Humanos , Sondas de Oligonucleótidos , Leucemia-Linfoma Linfoblástico de Células Precursoras/patología , Receptores de Antígenos de Linfocitos T gamma-deltaRESUMEN
T cell receptor delta (TCR) genes have been recently identified as rearranging during the early stages of T cell differentiation. We have analyzed the configuration of these genes in 47 unselected acute nonlymphoid leukemias. Morphology, phenotype, immunoglobulin heavy chain, and T cell receptor beta and gamma chain gene configuration were also studied. We have documented TCR delta gene rearrangements or deletions in eight cases using a genomic J delta 1 probe. The comparison of morphological, phenotypical, and molecular findings from these cases with those from control acute myeloid leukemias whose TCR delta genes were in germline configuration show that TCR delta rearrangements occur predominantly in immature leukemia exhibiting extensive lineage infidelity. The most striking feature was the frequent expression of the CD10 antigen. These data show that inappropriate gene rearrangements occur nonrandomly in myeloid leukemias and suggest that common mechanisms may be involved in the regulation of gene rearrangements and in the expression of some differentiation antigens.
Asunto(s)
Reordenamiento Génico de Linfocito T , Leucemia Mieloide Aguda/genética , Receptores de Antígenos de Linfocitos T/genética , Adolescente , Adulto , Antígenos CD/análisis , Diferenciación Celular , Preescolar , ADN Nucleotidilexotransferasa/análisis , Femenino , Genes de Inmunoglobulinas , Humanos , Leucemia Mieloide Aguda/inmunología , Masculino , Persona de Mediana Edad , Receptores de Antígenos de Linfocitos T gamma-deltaRESUMEN
Hairy cell leukemia (HCL) is a pre-plasma B cell tumor which responds to interferon (IFN)-alpha therapy. In vitro, B cell growth factor (BCGF) can induce proliferation of hairy cells. We have investigated the effect of in vitro and in vivo treatments with different recombinant IFN on the capacity of hairy cells to proliferate in response to human BCGF. In vitro treatment of leukemic cells from HCL patients with recombinant IFN-alpha-2 (5/5 cases) or IFN-beta (4/5 cases) resulted in a marked inhibition of the BCGF-dependent response. This suppressive effect was obtained with IFN concentrations of 1000, 100 IU/ml, and even occasionally 10 IU/ml. In contrast, no such inhibition was observed with IFN-gamma, despite the presence of specific IFN-gamma receptors on hairy cells at densities similar to receptors for IFN-alpha/beta. The IFN-alpha-induced suppression of the proliferative response of hairy cells to BCGF was also observed in vivo in two patients within 6-12 hr after administration of single doses of IFN-alpha. When hairy cells were maintained in culture for 1 week, they recovered their capacity to be stimulated by BCGF. This reversion was also shown in vivo in hairy cells isolated 1 week after IFN administration. Since in vivo growth of hairy cells could possibly result from the autocrine secretion of BCGF, we propose that the therapeutic effect of IFN-alpha on HCL may be due in part to an inhibition of such autocrine loop.
Asunto(s)
Interferones/farmacología , Interleucinas/farmacología , Leucemia de Células Pilosas/patología , 2',5'-Oligoadenilato Sintetasa/metabolismo , División Celular/efectos de los fármacos , Replicación del ADN/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Humanos , Interleucina-4 , Interleucinas/antagonistas & inhibidores , Receptores Inmunológicos/metabolismo , Receptores de Interferón , Factores de Tiempo , Células Tumorales CultivadasRESUMEN
Six patients with Philadelphia-positive (Ph1+) acute nonlymphocytic leukemia (ANLL) were studied by morphological, immunological, cytogenetic, and molecular techniques. Seven Ph1+ acute lymphocytic leukemia (ALL) cases were also studied for comparison. Three of ANLL cases were classified in M1, M2, and M4 groups of the FAB nomenclature, while the three other cases do not fit with any FAB subgroup and are described as M0. Immunophenotypical marker studies, double immunolabeling, and combined immunological and cytogenetic studies of metaphases showed that these ANLL expressed several lineage differentiation antigens. Rearrangements of immunoglobulin heavy chain gene (C mu) were detected in the six ANLL cases and in the seven ALL cases studied, as well as, in most cases, rearrangement of T cell receptor beta chain genes and/or T cell rearranging gamma genes. The results favored the assumption that the Ph1 translocation originated from a multipotent stem cell in Ph1+ ANLL. A common t(9;22) translocation was found in all cases, and additional chromosomal abnormalities were present in the six ANLL cases and in five of the seven ALL cases. Molecular studies of bcr gene configuration and c-abl transcription allowed two groups of Ph1+ ANLL to be distinguished. Three cases had bcr rearrangement and c-abl mRNA expression comparable to those reported in Ph1+ chronic myeloid leukemia, while three others had not detectable bcr rearrangement and a 7.2-7.5 kb c-abl mRNA. The existence of Ph1+ ALL with and without classical bcr rearrangement was confirmed.