Assuntos
Anemia/genética , Eliptocitose Hereditária/genética , Glucosefosfato Desidrogenase/genética , Mutação , Piruvato Quinase/genética , Anemia/diagnóstico , República Dominicana , Saúde da Família , Feminino , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Recém-Nascido , Padrões de Herança/genética , Masculino , Doenças Raras/diagnóstico , Doenças Raras/genética , Gêmeos Monozigóticos/genéticaRESUMO
We describe the finding of two Mexican patients with a specific 27-bp deletion in the solute carrier family 4 gene (SLC4A1delta27) (also known as the band 3 gene found on chromosome 17q21-q22), characteristic of Southeast Asian ovalocytosis (SAO). The patients were asymptomatic, and the initial diagnosis was made by microscopic observation of the presence of typical stomatocytic ovalocytes. The gene deletion was confirmed by PCR and DNA sequencing. Both patients were heterozygous for the deletion. One patient is from Tabasco state, in southeastern Mexico, a malaria-endemic zone. The other patient is from Mexico City, which is not a malaria-endemic area. Their families have no non-Mexican ancestors and their previous generations were born in Mexico. Both patients carry the HLA-B*3501 subtype, characteristic of Amerindians and Asian populations. Familial and HLA data led us to conclude that these two patients are the first report of SLC4A1delta27 in Amerindians. The nucleotide analysis showing a perfect match sequence between Southeast Asian and Mexican patients suggests, but does not prove, that the Mexican gene is not a de novo mutation. Instead, this gene might be the result of migration of individuals with Asian ancestry into the Mexican gene pool. We are looking for other families with the mutation to detect, by HLA analysis, the ancient ethnic origin of these patients.
Assuntos
Proteína 1 de Troca de Ânion do Eritrócito/genética , Eliptocitose Hereditária/genética , Deleção de Genes , Biologia Molecular , Sudeste Asiático , Sequência de Bases , Antígenos HLA-B/genética , Humanos , MéxicoRESUMO
Hereditary elliptocytosis (HE) is a group of hemolytic anemias characterized by the presence of elliptical erythrocytes. The underlying alterations lie in the proteins of the membrane skeleton. Defects of the alphaI domain of spectrin have been defined based on a decrease in the normal 80-kD alphaI domain and a concomitant increase in one or more lower molecular weight peptides. We have studied three Brazilian kindreds with black ancestry, who presented mild common spalphaI/50 HE. Our aim was to determine the molecular alteration responsible for the spalphaI/50 HE observed in these three kindreds and to evaluate the presence and influence of allele alphaLELY in the expression of this type of HE. In order to establish the molecular defect, exons 5, 6 and 11 were amplified and submitted to a nonradioactive single strand conformation polymorphism protocol. An identical band shift in exon 6 was observed in all 3 patients and their affected relatives. Direct sequencing of the amplification products of exon 6 showed the same molecular defect in all patients: a T-->C substitution, responsible for the L260P mutation. Allele alphaLELY, detected by PCR and restriction enzyme digestion, was present in the heterozygous form in the three propositi and was associated in trans with the elliptocytogenic mutation. Blood smears of the patients with HE and alphaLELY in trans showed pronounced elliptocytosis, poikilocytosis and a few small red cell fragments, whereas the blood smears of their relatives, who had HE without allele alphaLELY, showed mild common HE with a predominance of ovalocytes and the absence of poikilocytes. We conclude that allele alphaLELY does not lead to the worsening of clinical conditions when associated in trans with mild HE, but can be easily distinguished by a blood smear analysis. The predominance of the L260P mutation in the kindreds studied could be related to the colonization of Brazil during the slave trade by Africans from the Benin-Togo area, where this mutation is particularly common.
Assuntos
Alelos , Eliptocitose Hereditária/sangue , Eliptocitose Hereditária/genética , Espectrina/genética , Adolescente , Adulto , Brasil , Eliptocitose Hereditária/epidemiologia , Feminino , Humanos , Masculino , Linhagem , Espectrina/biossínteseRESUMO
Beta-Spectrin Campinas is a novel spectrin variant associated with a shortened beta-chain in a kindred with hereditary elliptocytosis (HE). The propositus and her mother exhibited increased amounts of spectrin dimers and an increase in the alphaI 74 kD fragment from the alpha-chain after partial tryptic digestion of spectrin. The shortened beta-chain appeared as an additional band of approximately 200 kD on SDS-PAGE. In order to delineate the molecular defect of this abnormality at the gene level, reticulocyte mRNA was transcribed into cDNA and the last four exons of the beta-spectrin gene were amplified. Agarose gel of the amplification product of the propositus revealed the expected band of 487 bp as well as a shortened band of approximately 300 bp (size determined on gel). This shortened cDNA amplification product was cloned and nucleotide sequencing revealed the absence of the entire exon 30. In order to determine the underlying mutation responsible for this abnormal splicing, a genomic DNA fragment containing exons 30 and 31 was amplified and nucleotide sequencing revealed a G-->A substitution at the 5' donor splice site consensus sequence of intron 30 (nt + 1 IVS30). The skip splicing observed in this study results in a frameshift, creating a new stop codon and causing a deletion of 129 amino acids at the very COOH-terminus of the protein, thus impairing spectrin dimers self-association. We classified this HE as spherocytic HE because the propositus presented a few spherocytes in addition to many elliptocytes in the blood smear, whereas her mother, who was splenectomized, showed many schizocytes, poikilocytes and spherocytes.
Assuntos
Eliptocitose Hereditária/genética , Éxons/genética , Mutação , Espectrina/genética , Adulto , Feminino , Regulação da Expressão Gênica , Humanos , Immunoblotting , Reação em Cadeia da Polimerase , Análise de Sequência de DNARESUMO
We report the clinical and laboratory findings in three unrelated families from southeastern Brazil with Sp alpha I/65 hereditary elliptocytosis (HE), including one homozygote and a patient presenting an elongated beta-spectrin. In family 1, three patients presented the allele alpha-Lely in trans to the elliptocytogenic allele. In these three patients the blood smear showed pronounced elliptocytosis, poikilocytosis and a few small red cell fragments instead of the mild elliptocytosis observed in their father, who did not present the polymorphism. In family 2 we describe one homozygote, with consanguineous parents presenting with anaemia, splenomegaly, severe poikilocytosis and elliptocytosis, budding, microspherocytes and numerous fragments in the blood smear. In family 3 we found an elongated beta Sp in a patient with Sp alpha I/65. The cause of the HE was the Sp alpha I/65 since the elongated beta Sp was not found in his brother, who also presented with HE and Sp alpha I/65. Apparently the abnormal beta Sp did not aggravate the HE, because both individuals had the same clinical and laboratory findings. However, the propositus presented a few more elliptocytes and poikilocytes than his brother, probably because the elongated beta-spectrin may have disturbed the spectrin self-association. In fact, in the propositus an abnormal band was observed in the nondenaturing gels, just above the Sp dimer, probably as a result of the association of the abnormal beta Sp with the normal spectrin chains. In the family studied here, both brothers presented the allele alpha Lely, but as their mother was dead, it was not possible to determine the polymorphism transmission. However, the high number of poikilocytes observed in the blood smear of both cases suggests an association in trans with the Sp alpha I/65. Thus, taken together, the data in this report indicate that HE secondary to Sp alpha I/65 abnormality is frequent in Brazil, and in one case it was associated with an apparently novel abnormal large beta-spectrin.
Assuntos
Eliptocitose Hereditária/metabolismo , Espectrina/metabolismo , Adulto , Brasil , Pré-Escolar , Eliptocitose Hereditária/genética , Feminino , Homozigoto , Humanos , Masculino , LinhagemAssuntos
Humanos , Proteínas de Membrana/química , Eliptocitose Hereditária/genética , Esferocitose Hereditária/genética , Membrana Eritrocítica/química , Espectrina/deficiência , Espectrina/genética , Espectrina/química , Anquirinas/genética , Eritrócitos/química , Proteínas de Membrana/genética , Deficiência de Proteína , Eliptocitose Hereditária/etiologia , Esferocitose Hereditária/etiologia , Membrana Eritrocítica/patologia , Mutação , /deficiência , /genéticaAssuntos
Eliptocitose Hereditária/genética , Membrana Eritrocítica/química , Proteínas de Membrana/química , Esferocitose Hereditária/genética , Proteína 1 de Troca de Ânion do Eritrócito/química , Proteína 1 de Troca de Ânion do Eritrócito/deficiência , Proteína 1 de Troca de Ânion do Eritrócito/genética , Anquirinas/genética , Eliptocitose Hereditária/etiologia , Eritrócitos/química , Glicoforinas/deficiência , Humanos , Proteínas de Membrana/genética , Mutação , Espectrina/química , Espectrina/deficiência , Espectrina/genética , Esferocitose Hereditária/etiologiaRESUMO
The Pelger-Huët anomaly (PHA) and the hereditary elliptocytosis (HE) are alterations affecting leukocytes and erythrocytes, respectively. Most of the affected individuals do not present clinic manifestations and are casually detected in the laboratory. The PHA and HE were described related to other hereditary and congenital conditions, but rarely have been found in the same individual. In this paper are reported discovery of the PHA and HE combined both in sister and brother, with global delay of development and peculiar physical characteristics. Blood smears of both showed an increase of bilobed neutrophils and elliptic erythrocytes. The family study showed two more members of the mother's branch affected with HE. The PHA could not be found in the parents, nor the other members of both branches. The early deaths or in uterus of three sibs of the propositus are appointed. The absence of the PHA in both parents of the affected individuals,--without discarding the illitimacy possibility--, can be explained by an incomplete genetic penetrance. The early or in uterus deaths of three sibs of the propositus seems possible that in some cases the combination of PHA and HE could be lethal. Possibly in Mexico this may be the first report of the presentation of the PHA and HE simultaneously in the same individual.
Assuntos
Eliptocitose Hereditária/complicações , Anomalia de Pelger-Huët/complicações , Adulto , Criança , Pré-Escolar , Eliptocitose Hereditária/genética , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Linhagem , Anomalia de Pelger-Huët/genéticaRESUMO
Se estudiaron nueve pacientes (cuatro familias) con eliptocitosis herediataria (EH). Desde el punto de vista clínico, se identificaron dos tipos de presentación: siete pacientes fueron formas compensadas y dos (de 7 y 63 años) pertenecientes a distintas familias mostraron anemia, esplenomegalia, reticulocitosis y fragmentación globular. En una familia se comprobó una disminución del stock total de la proteína 4.1 en la electroforesis de proteína de membrana en gel de poliacrilamida con SDS (SDS-PAGE). Uno de sus miembros se comprobó como forma descompensada. El estudio de dímeros y tetrámeros y la digestión tríptica de espectrina fue normal en todos los casos. En la determinación de la deformabilidad en función de la osmoralidad con el ectacitómetro se halló una buena correlación entre el índice de deformabilidad (ID) y el grado de anemia. De acuerdo a los resultados de este estudio pudimos confirmar la gran heterogeneidad clínica de la EH aún en miembros de una misma familia
Assuntos
Humanos , Masculino , Feminino , Pré-Escolar , Criança , Adulto , Idoso , Deformação Eritrocítica/genética , Eliptocitose Hereditária/genética , Proteínas de Membrana/deficiência , Eliptocitose Hereditária/sangue , Citometria de Fluxo , Membrana Eritrocítica/metabolismo , Espectrina/biossínteseRESUMO
Se estudiaron nueve pacientes (cuatro familias) con eliptocitosis herediataria (EH). Desde el punto de vista clínico, se identificaron dos tipos de presentación: siete pacientes fueron formas compensadas y dos (de 7 y 63 años) pertenecientes a distintas familias mostraron anemia, esplenomegalia, reticulocitosis y fragmentación globular. En una familia se comprobó una disminución del stock total de la proteína 4.1 en la electroforesis de proteína de membrana en gel de poliacrilamida con SDS (SDS-PAGE). Uno de sus miembros se comprobó como forma descompensada. El estudio de dímeros y tetrámeros y la digestión tríptica de espectrina fue normal en todos los casos. En la determinación de la deformabilidad en función de la osmoralidad con el ectacitómetro se halló una buena correlación entre el índice de deformabilidad (ID) y el grado de anemia. De acuerdo a los resultados de este estudio pudimos confirmar la gran heterogeneidad clínica de la EH aún en miembros de una misma familia (AU)