RESUMEN
BACKGROUND: The recessive X-linked disorder hemophilia A (HA) is rarely expressed in female carriers, most of whom express about half of normal factor VIII activity ( FVIII: C). OBJECTIVE: To propose an integrative assessment model for the binary role of the phase between the mutated F8 and the active X-chromosome (Xa) in FVIII: C in HA carriers. METHODS: We studied 67 females at risk of severe HA, comprising five symptomatic females ( FVIII: C < 1.5 IU dL(-1) ) and 14 controls. A correlation study between FVIII: C (observed vs. expected) and X-chromosome inactivation (XCI) patterns (XIPs; androgen receptor gene [AR] system) in blood leukocyte DNA was performed in carriers, by comparison of a model correlating FVIII: C and XIP with arbitrary models devoid of biological significance, and with FVIII: C levels in non-carriers (mean model) as a proxy from background data dispersion not influenced by XIP. RESULTS: We provide proof-of-concept example from a family presenting with extremely skewed XIPs in which the severe HA phenotype appeared in a heterozygous carrier of a crossover between AR and F8 loci that phased the mutated F8 with the maternally inherited Xa. Furthermore, four cases of severe HA affected women who had a combination of a heterozygous F8 mutation and extremely skewed XIPs in leukocytes or oral mucosa are presented. Correlation analyses between FVIII: C levels and XIPs in carriers (n = 38) but not in non-carriers (n = 20) showed highly significant differences between the proposed correlation model and models without biological significance. The data support a binary influence of XCI, either increasing or decreasing the FVIII: C, subject to the underlying phase set between the F8 mutation and XCI. CONCLUSIONS: Our evidence suggests that the phase between XCI and mutated F8 acts as a molecular switch conditioning FVIII: C levels and HA expression in carriers.
Asunto(s)
Cromosomas Humanos X , Factor VIII/genética , Hemofilia A/genética , Mutación , Inactivación del Cromosoma X , Adolescente , Adulto , Estudios de Casos y Controles , Niño , Preescolar , Factor VIII/análisis , Femenino , Estudios de Asociación Genética , Marcadores Genéticos , Predisposición Genética a la Enfermedad , Hemofilia A/sangre , Hemofilia A/diagnóstico , Herencia , Heterocigoto , Humanos , Lactante , Persona de Mediana Edad , Linaje , Fenotipo , Receptores Androgénicos/genética , Factores de Riesgo , Índice de Severidad de la Enfermedad , Adulto JovenRESUMEN
Inhibitor development against exogenous factor VIII is a severe impairment of replacement therapy affecting 18% of Argentine patients with severe haemophilia A (HA). To study the molecular predisposition for inhibitor development, we genotyped 260 HA patients with and without inhibitors, countrywide. The inhibitor-positive population (19 transients, 15 low responders, LR and 70 high responders, HR) of 104 severe-HA patients showed 59 Inv22 (intron 22 inversions), 18 small ins/del-frameshifts, 12 gross deletions, 12 nonsense, one splicing defect and two missense, p.Arg531Pro and p.Leu575Pro, both LR and thought to impair FVIII A2 domain secondary structure. In addition, a patient with mild HA and HR showed the missense p.Glu1704Lys associated with two neutral intronic substitutions potentially affecting the A3 domain. A case/control study (84/143) permitted estimation of F8 genotype-specific inhibitor risks [OR; prevalence (CI)] in severe-HA patients classifying a high-risk group including multi-exon deletions [3.66; 55% (19-100)], Inv22 [1.8; 24% (19-100)] and nonsense in FVIII-LCh [1.2; 21% (7-59)]; an average risk group including single-exon deletions, indel frameshifts and nonsense-HCh; and a low-risk group represented by missense defects [0.14; 3% (0.6-11)]. Analysis of inhibitor concordance/discordance in related patients indicated additional genetic factors other than F8 genotype for inhibitor formation. No significant inhibitor-predisposing factors related to FVIII product exposure were found in age- and F8 genotype-stratified populations of severe-HA patients. In conclusion, the Argentine HA patient series presents similar global and mutation-specific inhibitor risks than the HA database and other published series. This case-specific information will help in designing fitted therapies and follow-up protocols in Argentina.
Asunto(s)
Factor VIII/antagonistas & inhibidores , Factor VIII/genética , Predisposición Genética a la Enfermedad , Hemofilia A/genética , Argentina , Estudios de Casos y Controles , Humanos , Factores de RiesgoRESUMEN
Hepatitis C viraemia, in 38 human immunodeficiency virus positive (HIV+)/hepatitis C virus positive (HCV+) patients, was determined in haemophilic patients during the 4 years since initiation of highly active antiretroviral therapy (HAART). Six of 38 patients had persistently HCV-negative viraemia for more than 2 years. No correlation between HCV-negative viraemia and CD4+ T-cell counts, HIV viral load, age, type or severity of haemophilia could be established. Reduced levels of HIV viral load and the immune reconstitution that follows the initiation of HAART were not enough to explain the disappearance of HCV from plasma. Individuals who cleared plasma HCV had significantly higher CD8+ T-cell counts (P=0.0013) (mean +/- SE: 1153 +/- 117.8 cells microL(-1)) than those with HCV-positive viraemia (819.1 +/- 40.72 cells microL(-1)). Because HCV could maintain a low replication level in peripheral blood mononuclear cells (PBMC), we cultured PBMC of five of six patients with undetectable HCV viraemia. We found four of five HCV RNA-positive cultures. The presence of HCV RNA in our cultures proved that these cells may be an important viral reservoir that could contribute to HCV recurrence in plasma even after long periods of negative viraemia. In summary, our results indicate that in spite of prolonged HCV-negative plasma viraemia, HCV patients that are co-infected with HIV may harbour replication-competent HCV in their PBMC. Therefore, true clearance of HCV infection is difficult to achieve in these patients.
Asunto(s)
Infecciones por VIH/complicaciones , Hemofilia A/complicaciones , Hepacivirus/aislamiento & purificación , Hepatitis C/complicaciones , Leucocitos Mononucleares/virología , Adulto , Terapia Antirretroviral Altamente Activa , Recuento de Linfocito CD4 , Células Cultivadas , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/inmunología , Hemofilia B/complicaciones , Hepacivirus/fisiología , Hepatitis C/inmunología , Hepatitis C/virología , Humanos , Masculino , ARN Viral/análisis , Carga Viral , Viremia/complicaciones , Viremia/virología , Latencia del VirusRESUMEN
Primary cultures of peripheral blood mononuclear cells (PBMC) from 51 HIV+ hemophiliac patients (HIV+ PBMC) were set up, allowing undisturbed cellular interaction in the absence of any exogenous stimuli. The optimum time for p24 detection was between 12 and 25 days. Infective virus was recovered from the culture supernatants (HIV+ SN) and the amount of p24 released ranged from 25 to 5300 pg/ml. Cells of the monocyte/macrophage (M/M) lineage were the main source of HIV in the HIV+ SN, as judged by intracellular staining of permeabilized cells with anti-p24 (KC57 monoclonal antibody) and flow cytometry analysis. M/M activation, differentiation, and proliferation occurred along the culture before the peak of in vitro HIV replication. Release of HIV p24 was highest in patients with >200 CD4+ T lymphocytes/mm3 who did not receive highly active antiretroviral therapy (HAART), but it was still detectable in 60-90% of patients who had responded to 1-2 years of HAART, reducing their plasma viral load to undetectable levels. It is proposed that this simple experimental system can be used to assess ongoing HIV infection of M/M with the patient's own viral variants.
Asunto(s)
Técnicas de Cultivo de Célula/métodos , Infecciones por VIH/virología , VIH/aislamiento & purificación , Monocitos/virología , Terapia Antirretroviral Altamente Activa , Diferenciación Celular , División Celular , Células Cultivadas , Medios de Cultivo Condicionados/química , Femenino , VIH/crecimiento & desarrollo , Proteína p24 del Núcleo del VIH/análisis , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/inmunología , Humanos , Antígeno Ki-67/análisis , Cinética , Macrófagos/química , Macrófagos/citología , Macrófagos/virología , Masculino , Monocitos/química , Monocitos/citología , Replicación ViralRESUMEN
Intron 22 factor VIII gene inversion (Inv22) is the most common mutation causing severe haemophilia A (SHA). We studied Inv22 in 34 SHA affected families by Southern blotting. Data from the familial history of the disease and the inhibitor status were also included. We found Inv22 in 41 % of SHA Argentine families (35 % with type 1 and 6 % with type 2), in close agreement with previously reported series. No significant correlation between the inheritance (familiar or sporadic disease) and the presence of inversions was found. Our population showed 24 % of families included at least one hemophiliac with inhibitor. In families positive for Inv22, 29 % of patients developed inhibitor but this increased frequency was not statistically significant. In conclusion, analysis of Inv22 in SHA patients should be used as a first line method because it provides useful and secure information for carrier detection and prenatal diagnosis in a high percentage of cases.
Asunto(s)
Inversión Cromosómica , Factor VIII/genética , Hemofilia A/genética , Intrones/genética , Argentina/epidemiología , Southern Blotting , Sondas de ADN , Factor VIII/inmunología , Salud de la Familia , Femenino , Frecuencia de los Genes , Hemofilia A/epidemiología , Hemofilia A/inmunología , Humanos , Isoanticuerpos/sangre , Masculino , Polimorfismo de Longitud del Fragmento de RestricciónRESUMEN
The last 40 years have witnessed important changes in the understanding and treatment of oncohematological affections. Palliative therapy was gradually replaced by chemotherapy (CT) which rapidly proved unexpectedly effective. In 1948, the first antifolic drugs, aminopterin and methotrexate, were discovered, followed in 1950 by the corticoids and in 1953 by antipurine agents. By 1967, a combination of these drugs yielded a survival index of 50% in acute lymphoblastic leukemia (ALL) with a progressive increase in all important cancer centers today, including in GATLA (Argentine Group for Acute Leukemia). As for acute myeloblastic leukemia (AML) the CT results were not as spectacular although now there is a 25% survival index which reaches 40-50% in young adults. As for allogeneic transplant in acute leukemia, its use must be evaluated for each patient and for each circumstance. Leukemias are genetic diseases for which gene therapy undoubtedly has potential value. However, the problems raised by the election of the right gene or gene marker and specially of the adequate vector have not yet been solved. In Hodgkin's disease, the results obtained with CT since the decade of the 60s have been spectacular and today different combinations of drugs have yielded a survival rate above 80%. Immunotherapy with or without CT has opened up a completely new and promising field. The route from basic research to clinical application has been long and arduous but the results obtained in leukemia and lymphomas have undoubtedly been life-saving and hopefully will open up even better possibilities in the near future.
Asunto(s)
Leucemia Linfoide/terapia , Oncología Médica/tendencias , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Trasplante de Médula Ósea , Supervivencia sin Enfermedad , Predicción , Terapia Genética , Humanos , InmunoterapiaRESUMEN
The last 40 years have witnessed important changes in the understanding and treatment of oncohematological affections. Palliative therapy was gradually replaced by chemotherapy (CT) which rapidly proved unexpectedly effective. In 1948, the first antifolic drugs, aminopterin and methotrexate, were discovered, followed in 1950 by the corticoids and in 1953 by antipurine agents. By 1967, a combination of these drugs yielded a survival index of 50
in acute lymphoblastic leukemia (ALL) with a progressive increase in all important cancer centers today, including in GATLA (Argentine Group for Acute Leukemia). As for acute myeloblastic leukemia (AML) the CT results were not as spectacular although now there is a 25
survival index which reaches 40-50
in young adults. As for allogeneic transplant in acute leukemia, its use must be evaluated for each patient and for each circumstance. Leukemias are genetic diseases for which gene therapy undoubtedly has potential value. However, the problems raised by the election of the right gene or gene marker and specially of the adequate vector have not yet been solved. In Hodgkins disease, the results obtained with CT since the decade of the 60s have been spectacular and today different combinations of drugs have yielded a survival rate above 80
. Immunotherapy with or without CT has opened up a completely new and promising field. The route from basic research to clinical application has been long and arduous but the results obtained in leukemia and lymphomas have undoubtedly been life-saving and hopefully will open up even better possibilities in the near future.
Asunto(s)
Infecciones por VIH/transmisión , Hemofilia A/terapia , Reacción a la Transfusión , Adolescente , Adulto , Factores de Edad , Anciano , Niño , Preescolar , Femenino , Infecciones por VIH/epidemiología , Infecciones por VIH/etiología , Humanos , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Prevalencia , Factores de Riesgo , Perfil de Impacto de EnfermedadRESUMEN
Mixed, bilineal, biclonal and hybrid leukemias are synonymous, differing from biphenotypical ones. Mixed acute leukemia is defined by the coincidence of 1) two cytochemical markers of different lineage, or 2) one of them with more than one opposite immunological marker, or 3) more than one immunological marker opposite to another immunological lineage. Seven cases of mixed acute leukemia are presented, two of which showed posttreatment switching. It is concluded that mixed acute leukemias are associated with a poor prognosis, and therapeutic criteria are defined.
Asunto(s)
Leucemia/clasificación , Leucemia/inmunología , Enfermedad Aguda , Adolescente , Adulto , Anciano , Línea Celular , Niño , Preescolar , Femenino , Humanos , Leucemia/terapia , Leucemia Bifenotípica Aguda/inmunología , Leucemia Bifenotípica Aguda/terapia , Masculino , PronósticoRESUMEN
Between 1960 and 1991, 156 episodes of central nervous system (CNS) bleeding were documented in 106 patients from a total population of 1,410 hemophiliacs (7.5%). Ninety-one hemophilia A patients presented 131 bleeding episodes; 15 hemophilia B patients had 25 episodes. 32% of these episodes took place in patients less than 5 years of age. 46% were age 10 or less, and 72% were age 20 or less. The mean age was 14.8 years in hemophilia A and 9 years in hemophilia B patients. A significant increase in the mean age of hemophilia A patients has been observed over the last 10 years; this may be related to HIV infection. A history of recent trauma was documented in 39.7% of the episodes. Spontaneous CNS bleeding was predominant in severe hemophilia (85.2%). One hundred and fifty-four CNS bleeding episodes were intracranial and 2 intraspinal. Of the intracranial episodes, 37.7% were subarachnoid, 29.8 subdural, and 22.7% intracerebral. Factor VIII or IX inhibitors were present in 11.3% of the patients; this figure is slightly lower than that observed in our total hemophilic population. Over 50% of the patients had psychoneurological sequelae; the most frequent were seizure disorders and motor impairment. The overall mortality rate was 29.2%. The mortality was more closely related to the CNS bleeding site than to the severity of hemophilia. Treatment should be based on prompt and prolonged replacement therapy to ensure hemostatic levels of antihemophilia factors.
Asunto(s)
Hemorragia Cerebral/epidemiología , Hematoma Subdural/epidemiología , Hemofilia A/complicaciones , Hemorragia Subaracnoidea/epidemiología , Adolescente , Adulto , Factores de Edad , Anciano , Hemorragia Cerebral/etiología , Niño , Preescolar , Traumatismos Craneocerebrales/complicaciones , Femenino , Infecciones por VIH/complicaciones , Hematoma Subdural/etiología , Hematoma Subdural/cirugía , Humanos , Incidencia , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Recurrencia , Rotura Espontánea , Convulsiones/etiología , España/epidemiología , Enfermedades de la Columna Vertebral/epidemiología , Enfermedades de la Columna Vertebral/etiología , Hemorragia Subaracnoidea/etiología , Tasa de Supervivencia , Resultado del TratamientoRESUMEN
We present studies on the evolution of HIV-1 infection in 638 hemophilic patients receiving commercial antihemophilic concentrates (CAH) at the Institute of Hematological Research and the Argentine Foundation of Hemophilia between 1983 and 1990. Positive serology for HIV-1 was detected in 30% of the patients studied. Prevalence of HIV-1 infection was higher (about 70%) in the group with severe hemophilia requiring more CAH, but there were no differences between patients with hemophilia A or B. Sexual transmission was demonstrated in 8/64 women (13%) with stable sexual relationship with HIV-1 + hemophilic patients. Three of them became pregnant, and HIV-1 infection was demonstrated in two of the three children. In general, the clinical evolution, as well as the hematologic and immunologic parameters of infected patients were similar to those described for the hemophilic population in other occidental countries. Opportunistic infections were also those observed elsewhere (with predominance of P. carinii pneumonia and disseminated Candida infections). However, the presence of fatal chagasic encephalitis in two of the patients with AIDS is unusual. Thus, central nervous system localization of T. cruzi (which can be observed during the acute period of T. cruzi infection or in immunosuppressed patients), must be considered as a possible severe complication of HIV-1 disease in T. cruzi infected patients.
Asunto(s)
Síndrome de Inmunodeficiencia Adquirida/complicaciones , Seroprevalencia de VIH , Hemofilia A/complicaciones , Infecciones Oportunistas Relacionadas con el SIDA/complicaciones , Síndrome de Inmunodeficiencia Adquirida/epidemiología , Síndrome de Inmunodeficiencia Adquirida/transmisión , Argentina/epidemiología , Enfermedad de Chagas/complicaciones , Encefalitis/complicaciones , Femenino , Seropositividad para VIH/diagnóstico , Hepatitis/complicaciones , Humanos , Masculino , Embarazo , Complicaciones Infecciosas del EmbarazoRESUMEN
We present studies on the evolution of HIV-1 infection in 638 hemophilic patients receiving commercial antihemophilic concentrates (CAH) at the Institute of Hematological Research and the Argentine Foundation of Hemophilia between 1983 and 1990. Positive serology for HIV-1 was detected in 30
of the patients studied. Prevalence of HIV-1 infection was higher (about 70
) in the group with severe hemophilia requiring more CAH, but there were no differences between patients with hemophilia A or B. Sexual transmission was demonstrated in 8/64 women (13
) with stable sexual relationship with HIV-1 + hemophilic patients. Three of them became pregnant, and HIV-1 infection was demonstrated in two of the three children. In general, the clinical evolution, as well as the hematologic and immunologic parameters of infected patients were similar to those described for the hemophilic population in other occidental countries. Opportunistic infections were also those observed elsewhere (with predominance of P. carinii pneumonia and disseminated Candida infections). However, the presence of fatal chagasic encephalitis in two of the patients with AIDS is unusual. Thus, central nervous system localization of T. cruzi (which can be observed during the acute period of T. cruzi infection or in immunosuppressed patients), must be considered as a possible severe complication of HIV-1 disease in T. cruzi infected patients.
RESUMEN
We present studies on the evolution of HIV-1 infection in 638 hemophilic patients receiving commercial antihemophilic concentrates (CAH) at the Institute of Hematological Research and the Argentine Foundation of Hemophilia between 1983 and 1990. Positive serology for HIV-1 was detected in 30
of the patients studied. Prevalence of HIV-1 infection was higher (about 70
) in the group with severe hemophilia requiring more CAH, but there were no differences between patients with hemophilia A or B. Sexual transmission was demonstrated in 8/64 women (13
) with stable sexual relationship with HIV-1 + hemophilic patients. Three of them became pregnant, and HIV-1 infection was demonstrated in two of the three children. In general, the clinical evolution, as well as the hematologic and immunologic parameters of infected patients were similar to those described for the hemophilic population in other occidental countries. Opportunistic infections were also those observed elsewhere (with predominance of P. carinii pneumonia and disseminated Candida infections). However, the presence of fatal chagasic encephalitis in two of the patients with AIDS is unusual. Thus, central nervous system localization of T. cruzi (which can be observed during the acute period of T. cruzi infection or in immunosuppressed patients), must be considered as a possible severe complication of HIV-1 disease in T. cruzi infected patients.
RESUMEN
Concanavalin A (Con-A)-induced suppression of T cell proliferation was studied in 48 patients with severe hemophilia. Two groups of patients were defined according to the proliferative response when increasing numbers of Con A-induced cells were added to a constant number of phytohemagglutinin (PHA)-stimulated autologous T cells: In group A (60%) and in normal controls, higher suppression was achieved when more Con A-induced cells were added; in Group B, increasing numbers of Con A-induced cells produced no suppression of stimulated PHA-triggered proliferation. This effect could be corrected in Group B by inducing suppression in the presence of inhibitors of the oxidative metabolism of arachidonic acid. No correlation was found between the suppression profile and HIV-1 or HBV serology. Clinical evolution, as judged by signs and symptoms of AIDS related complex tended to be better in Group B than in Group A patients. It is suggested that decreased Con A-induced suppression in Group B may represent part of a normal regulatory process that involves products of arachidonic acid oxidative metabolism.
Asunto(s)
Concanavalina A , Hemofilia A/inmunología , Activación de Linfocitos/efectos de los fármacos , Linfocitos T/efectos de los fármacos , Adolescente , Adulto , Seropositividad para VIH/inmunología , Humanos , Linfocitos T/inmunologíaRESUMEN
Concanavalin A (Con-A)-induced suppression of T cell proliferation was studied in 48 patients with severe hemophilia. Two groups of patients were defined according to the proliferative response when increasing numbers of Con A-induced cells were added to a constant number of phytohemagglutinin (PHA)-stimulated autologous T cells: In group A (60
) and in normal controls, higher suppression was achieved when more Con A-induced cells were added; in Group B, increasing numbers of Con A-induced cells produced no suppression of stimulated PHA-triggered proliferation. This effect could be corrected in Group B by inducing suppression in the presence of inhibitors of the oxidative metabolism of arachidonic acid. No correlation was found between the suppression profile and HIV-1 or HBV serology. Clinical evolution, as judged by signs and symptoms of AIDS related complex tended to be better in Group B than in Group A patients. It is suggested that decreased Con A-induced suppression in Group B may represent part of a normal regulatory process that involves products of arachidonic acid oxidative metabolism.
RESUMEN
An update of two consecutive randomized studies in previously untreated multiple myeloma was performed. The first study (10-M-73) began in 1973; 150 patients were treated with melphalan and prednisone (MP) or semustine, cyclophosphamide, and prednisone (MeCP). In a second randomized study (3-M-77), begun in 1977, 260 patients were treated with MP or melphalan, prednisone, cyclophosphamide, semustine, and vincristine (MPCCV). A total of 27 of the 67 patients (40%) treated with MP and 33 of the 83 patients (40%) treated with MeCP showed a good response in protocol 10-M-73; 48 of 145 patients (33%) treated with MP and 51 of the 115 patients (44%) treated with MPCCV in protocol 3-M-77 obtained a good response (P is not significant). Median survival in protocol 10-M-73 was 30 months for MeCP and 38 months for MP. At 84 months, 19% and 9% remain alive, respectively. Median survival for protocol 3-M-77 was 44 months for those treated with MPCCV and 42 months for MP. At 60 months, 9% and 11% remain alive; this difference was not significant. Also, there was no survival difference for favorable or unfavorable prognostic groups among the four treatment arms of both protocols. It can be concluded, with a long-term follow-up of both protocols, that the combination of MP is as effective as the three- and five-drugs combinations, and in view of its simplicity and cost-saving advantages, it should be favored for initial therapy of multiple myeloma patients.