RESUMEN
BACKGROUND: Monitoring of peripheral blood lymphocyte subpopulation (PBLS) counts might be useful for estimating the risk of infection after liver transplantation (LT). METHODS: We prospectively measured total lymphocyte and PBLS counts at baseline and post-transplant months 1 and 6 in 92 LT recipients. PBLS were enumerated by single-platform 6-color flow cytometry technology. Areas under receiver operating characteristic (ROC) curves were used to evaluate the accuracy of different PBLS for predicting cytomegalovirus (CMV) disease and overall opportunistic infection (OI). Adjusted hazard ratios (aHRs) for both outcomes were estimated by Cox regression. RESULTS: After a median follow-up of 730.0 days, 29 patients (31.5%) developed 38 episodes of OI (including 22 episodes of CMV disease). The counts of CD3(+) , CD4(+) , and CD8(+) T cells, and CD56(+) CD16(+) natural killer (NK) cells at month 1 were significantly lower in patients subsequently developing OI. The NK cell count was the best predictive parameter (area under ROC curve for predicting CMV disease: 0.78; P-value = 0.001). Patients with an NK cell count <0.050 × 10(3) cells/µL had higher cumulative incidences of CMV disease (P-value = 0.001) and overall OI (P-value <0.001). In the multivariate models, an NK cell count <0.050 × 10(3) cells/µL at month 1 post transplantation remained as an independent risk factor for CMV disease (aHR: 5.54; P-value = 0.003) and overall OI (aHR: 7.56; P-value <0.001). CONCLUSION: Post-transplant kinetics of NK cell counts may be used as a simple and affordable proxy to the cell-mediated immunity status in LT recipients and to their associated risk of OI.
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Infecciones por Citomegalovirus/sangre , Células Asesinas Naturales/inmunología , Trasplante de Hígado/efectos adversos , Subgrupos Linfocitarios/inmunología , Monitorización Inmunológica/métodos , Infecciones Oportunistas/sangre , Anciano , Citomegalovirus/aislamiento & purificación , Infecciones por Citomegalovirus/epidemiología , Infecciones por Citomegalovirus/inmunología , Infecciones por Citomegalovirus/virología , Femenino , Citometría de Flujo , Estudios de Seguimiento , Humanos , Inmunidad Celular , Recuento de Linfocitos/economía , Masculino , Persona de Mediana Edad , Infecciones Oportunistas/epidemiología , Infecciones Oportunistas/inmunología , Infecciones Oportunistas/microbiología , Valor Predictivo de las Pruebas , Estudios Prospectivos , Factores de RiesgoAsunto(s)
Anticuerpos Antibacterianos/inmunología , Formación de Anticuerpos/inmunología , Síndromes de Inmunodeficiencia/inmunología , Polisacáridos Bacterianos/inmunología , Salmonella typhi/inmunología , Vacunas Tifoides-Paratifoides/inmunología , Adulto , Agammaglobulinemia/inmunología , Anciano , Inmunodeficiencia Variable Común/inmunología , Ensayo de Inmunoadsorción Enzimática , Femenino , Interacciones Huésped-Patógeno/efectos de los fármacos , Interacciones Huésped-Patógeno/inmunología , Humanos , Inmunoglobulina G/inmunología , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Salmonella typhi/fisiología , Fiebre Tifoidea/inmunología , Fiebre Tifoidea/microbiología , Fiebre Tifoidea/prevención & control , Vacunas Tifoides-Paratifoides/administración & dosificación , Vacunación/métodos , Adulto JovenRESUMEN
APDS2 is caused by mutations in PIK3R1 gene resulting in constitutive PI3Kδ activation. PI3Kδ is predominantly expressed in leukocytes and plays critical roles in regulating immune responses. Here we first derived fibroblast primary cells from a skin biopsy of a patient carrying a heterozygous single T deletion in intron 11 of the PIK3R1 gene. We next present the derivation of an induced pluripotent stem cell (iPS) line using a non-integrative reprogramming technology. Pluripotent-related hallmarks are further shown, including: iPSCs self-renewal and expression of pluripotent and differentiation markers after in vitro differentiation towards embryonic germ layers, assessed by RT-PCR and immunofluorescence.
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Línea Celular , Células Madre Pluripotentes Inducidas , Enfermedades de Inmunodeficiencia Primaria/genética , Diferenciación Celular , Fosfatidilinositol 3-Quinasa Clase I/genética , Fibroblastos , Humanos , MutaciónRESUMEN
The purpose of this study is to evaluate the effects of neonatal thymectomy in the functional capacity of the immune system. We selected a group of 23 subjects, who had undergone thymectomy in their first 30 days of life, during an intervention for congenital heart disease. Several parameters of the immune system were evaluated during their first 3 years of life. Lymphocyte populations and subpopulations (including naive, memory and effector subpopulations), T cell receptor (TCR) Vbeta repertoire, response of T cells following in vitro stimulation by mitogen, quantification of immunoglobulins, TCR excision circles (TRECS) and interleukin (IL)-7 were measured. We found that neonatal thymectomy produces long-term diminution in total lymphocyte counts, especially in naive CD4+ and CD8+ T cells. Additionally, TRECS were decreased, and plasma IL-7 levels increased. A statistically significant negative correlation was found between absolute CD4+ T cells and IL-7 (r = -0.470, P = 0.02). The patients did not suffer more infectious events than healthy control children, but thymectomy in neonates resulted in a significant decrease in T lymphocyte levels and TRECS, consistent with cessation of thymopoiesis. This could produce a compromise in immune function later in life, especially if the patients suffer T cell depletion and need a reconstitution of immune function.
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Cardiopatías Congénitas/cirugía , Subgrupos de Linfocitos T/inmunología , Timectomía , Estudios de Seguimiento , Reordenamiento Génico de Linfocito T/inmunología , Humanos , Inmunidad Celular , Inmunoglobulinas/sangre , Inmunofenotipificación , Recién Nacido , Interleucina-7/sangre , Recuento de Linfocitos , Linfopenia/inmunología , Periodo Posoperatorio , Receptores de Antígenos de Linfocitos T/genética , Timo/inmunologíaRESUMEN
Papillon-Lefèvre syndrome (PLS) has recently been shown to be caused by mutations in the cathepsin C gene resulting in periodontal disease and palmoplantar keratosis. Thirteen different homozygous mutations have been characterised in PLS patients of different ethnic origin. In the present paper, a PLS patient is described who carries two novel mutations (706G>T and 872G>A) in the paternal and maternal chromosomes, respectively. This is the first compound patient described so far. In addition, a novel symptomless mutation (458C>T) in the cathepsin C gene is described in three homozygous individuals. Thus, not all mutations should be considered as a cause of disease, whether case studies or general population screening is performed. Another already described mutation that provoked the Haim-Munk syndrome (HMS) in Indian Jews has also been found to give rise to PLS in a Spanish family from Madrid. On the other hand, PLS patients are ameliorated by retinoids, which indicates that retinoids may be used as therapeutic agents in this immune system deficiency.
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Catepsina C/genética , Enfermedad de Papillon-Lefevre/genética , Alelos , ADN/química , ADN/genética , Análisis Mutacional de ADN , Salud de la Familia , Femenino , Frecuencia de los Genes , Heterocigoto , Humanos , Masculino , Datos de Secuencia Molecular , Mutación , Mutación Missense , Enfermedad de Papillon-Lefevre/enzimología , Enfermedad de Papillon-Lefevre/patologíaRESUMEN
The characterization of T cell immunodeficiencies could in part be supported by using stable cell lines in which biochemical and molecular studies of the defect could be carried out thereby omitting frequent bleeding of patients. First attempts to obtain such cell lines included HTLV-I transformation and exogenous IL-2 administration, but both models have important disadvantages. Recently, a virus isolated from the squirrel monkey, Herpes virus saimiri (HVS), has been reported to have the ability to transform T cells. A stable IL-2-dependent HVS-transformed T cell line from a CD3 gamma deficient patient has been obtained; and this cell line displays both the phenotypic and the functional characteristics of the patient's lymphocytes. Moreover, the line down-modulates TCR/CD3 surface expression upon CD3 engagement, as do the patient's lymphocytes, showing that CD3 gamma and its phosphorylation are not necessary for TCR/CD3 internalization. In addition, the abnormal staining pattern of different anti-TCR/CD3 monoclonal antibodies is preserved in the HVS-patient line. Since HVS is capable of transforming CD3 gamma- T cells, the CD3 gamma chain does not seem to be involved in the HVS receptor process. The fact that it is not possible to obtain a CD8+ HVS line from the CD3 gamma- patient supports the existence of a functional anomaly in his scanty CD8+ peripheral lymphocytes. Thus, HVS transformation is a suitable model for T cell immunodeficiency studies and characterization. It may also be used in the future in cellular models for in vitro gene therapy trials.
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Complejo CD3/análisis , Transformación Celular Viral/inmunología , Herpesvirus Saimiriino 2/inmunología , Síndromes de Inmunodeficiencia/inmunología , Activación de Linfocitos , Linfocitos T/inmunología , Linfocitos T/virología , Complejo CD3/genética , Línea Celular Transformada , Citometría de Flujo , Humanos , InmunofenotipificaciónRESUMEN
Six different ape MHC-G DNA sequences (four in humans: HLA-G*01011, HLA-G*II, HLA-G*0103, and HLA-G*IV; one in chimpanzees: Patr-G*I; and one in gorillas: Gogo-G*1) have been obtained. Only synonymous or conservative ("Thr"-to-"Ser") substitutions are allowed between the four human alleles. One allele of MHC-G exon-2 sequences has been found both in gorilla (Gorilla gorilla) and chimpanzee (Pan troglodytes). The Patr-G*I DNA sequence shows two nonsynonymous substitutions when compared with the human HLA-G*01011 sequence: "CGG"-to-"TGG" ("Arg"-to-"Trp") at codon 35 and "ATG"-to-"ATA" ("Met"-to-"Ile") at codon 76. One nonsynonymous "GAG"-to-"GGG" ("Glu"-to-"Gly") substitution is observed in the Gogo-G*I exon-2 DNA sequence, when compared with the human *01011 allele. None of these three different substitutions have been observed in humans and are, thus, considered species specific. Also, evidence is provided that the human HLA-G*II and G*0103 may have been originated after human speciation. Finally, phylogenetic relationships among the six MHC-G alleles, tamarins G-"like" alleles, and other human class I genes (both "classical" and "nonclassic") are discussed.
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Alelos , Genes MHC Clase I/genética , Primates/inmunología , Animales , Secuencia de Bases , Línea Celular , Gorilla gorilla , Antígenos HLA/genética , Antígenos HLA-G , Antígenos de Histocompatibilidad Clase I/genética , Humanos , Datos de Secuencia Molecular , Pan troglodytes , Filogenia , Primates/genética , Especificidad de la EspecieRESUMEN
BACKGROUND: Chronic Granulomatous Disease (CGD) is a rare primary immunodeficiency caused by the alteration of the enzyme complex NADPH oxidase, which affects the phagocytic function. CGD patients are susceptible to recurrent infections mainly caused by bacteria and/or fungi. METHODS: We studied a 6 year-old boy with suspicion of CGD. The diagnosis was confirmed based on the functional study of NADPH oxidase. Simultaneously, the second pregnancy of the mother was reported and genetic counselling was requested. RESULTS: We identified a new disease-causing mutation by direct sequencing of the CYBB gene (X-linked CGD). The prenatal study resulted in the identification of the same mutation in the foetus. COMMENTS: Molecular genetics characterisation of CGD is needed to obtain an accurate diagnosis of the disease and to offer prenatal diagnosis and genetic counselling in future pregnancies.
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Amniocentesis , Enfermedad Granulomatosa Crónica/diagnóstico , Niño , Femenino , Enfermedad Granulomatosa Crónica/genética , Humanos , Masculino , Mutación , EmbarazoRESUMEN
BACKGROUND: The autoimmune lymphoproliferative syndrome (ALPS) is caused by genetic defect in lymphocyte apoptosis. Chronic lymphadenopathy and splenomegaly are the consequence of lymphoproliferation. The diagnosis is based on the assessment of the defective lymphocyte apoptosis and the identification of lymphocyte T subset that are double negative (CD4-CD8-). The susceptibility to lymphoma and autoimmune diseases, mainly blood cytopenias is increased. METHODS: We studied a 14 year-old boy with chronic splenomegaly and familial history of splenomegaly and lymphadenopathy. T lymphocyte phenotypes, and molecular defect of TNFRSF6 gene were studied in the child, his sister and his father. Lymphocyte apoptosis was also analysed in the child and his father. RESULTS: The boy and his father showed in vitro apoptosis defects, an increased number of double negative T lymphocytes (18% and 5%, respectively) and the same mutation in the TNFRSF6 gene. His sister had 16% of double negative T lymphocytes and the mutation in the TNFRSF6 gene. COMMENTS: Chronic familial splenomegaly can be the only clinical sign of autoimmune lymphoproliferative syndrome.
Asunto(s)
Síndrome Linfoproliferativo Autoinmune/inducido químicamente , Síndrome Linfoproliferativo Autoinmune/complicaciones , Esplenomegalia/etiología , Esplenomegalia/genética , Adolescente , Humanos , Masculino , LinajeAsunto(s)
Trasplante de Médula Ósea , Genes RAG-1 , Inmunodeficiencia Combinada Grave/genética , Inmunodeficiencia Combinada Grave/terapia , Sustitución de Aminoácidos , Trasplante de Médula Ósea/inmunología , Niño , Preescolar , Estudios de Seguimiento , Heterocigoto , Humanos , Deficiencia de IgG/inmunología , Inmunoglobulina G/sangre , Lactante , Masculino , Mutación Puntual , Inmunodeficiencia Combinada Grave/inmunologíaRESUMEN
The phylogenetic relationships of genus Passer (Old World sparrows) have been studied with species covering their complete world living range. Mitochondrial (mt) cyt b genes and pseudogenes have been analyzed, the latter being strikingly abundant in genus Passer compared with other studied songbirds. The significance of these Passer pseudogenes is presently unclear. The mechanisms by which mt cyt b genes become pseudogenes after nuclear translocation are discussed together with their mode of evolution, i.e., transition/transversion mitochondrial ratio is decreased in the nucleus, as is the constraint for variability at the three codon positions. However, the skewed base composition according to codon position (in 1st position the percentage is very similar for the four bases, in 2nd position there are fewer percentage of A and G and more percentage of T, and in 3rd codon position fewer percentage of G and T and is very rich in A and C) is maintained in the translocated nuclear pseudogenes. Different nuclear internal mechanisms and/or selective pressures must exist for explaining this nuclear/mitochondrial differential DNA base evolutive variability. Also, the phylogenetic usefulness of pseudogenes for defining relationships between closely related lineages is stressed. The analyses suggest that the primitive genus Passer species comes from Africa, the Cape sparrow being the oldest: P. hispaniolensis italiae is more likely conspecific to P. domesticus than to P. hispaniolensis. Also, Passer species are not included within weavers or Estrildinae or Emberizinae, as previously suggested. European and American Emberizinae sparrows are closely related to each other and seem to be the earliest species that radiated among the studied songbirds (all in the Miocene Epoch).
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ADN Mitocondrial/genética , Filogenia , Seudogenes/genética , Pájaros Cantores/genética , Núcleo Celular/genética , Grupo Citocromo b/genética , ADN/química , ADN/genética , Evolución Molecular , Variación Genética , Datos de Secuencia Molecular , Análisis de Secuencia de ADN , Pájaros Cantores/clasificaciónRESUMEN
Mitochondrial cytochrome b (cyt b) from 24 Carduelini species including crossbills, bullfinches, grosbeaks, rosefinches, and other related, but not conclusively classified species, was sequenced. These sequences were also compared with all the available sequences from the genera Carduelis, Serinus, and Passer. Phylogenetic analyses consistently gave the same groups of finches and the calculated divergence times suggest that speciation of the studied species occurred between 14 and 3 million years ago (Miocene-Pliocene), appearing before the Passer, Carduelis, and Serinus genera. Pleistocene glaciations may have been important in sub-speciation. Crossbills are integrated within the genus Carduelis, and within redpolls; the common crossbill shows subspeciation with Loxia japonica in the Pleistocene epoch. Pinicola enucleator groups together with bullfinches and is probably the ancestor of the group. Hawfinch is only distantly related to the studied groups, and might either represent an isolated genus or be related to the New World genus Hesperiphona. The grosbeak genera Eophona and Mycerobas are clearly sister groups, and species belonging to the former might have given rise to Mycerobas species. The isolated (in classification) Uragus sibiricus and Haematospiza sipahi are included within the genus Carpodacus (rosefinches); Carpodacus nipalensis is outside the genus Carpodacus in the molecular analyses and might be an isolated species or related to the genus Montifringilla.
Asunto(s)
Grupo Citocromo b/genética , ADN Mitocondrial , Pájaros Cantores/clasificación , Sustitución de Aminoácidos , Animales , ADN , Filogenia , Pájaros Cantores/genéticaRESUMEN
The study of the MHC-G gene evolution during nearly 40 million years does not support a role for the full molecule. The MHC-G-like proteins of New World monkeys are probably classical presenting molecules. Old World Cercopithecinae monkeys do not have a full MHC-G molecule and human individuals homozygous for the HLA-G null allele are healthy and do not show birth pathologies.
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Genes MHC Clase I , Antígenos HLA/fisiología , Antígenos de Histocompatibilidad Clase I/fisiología , Secuencia de Aminoácidos , Animales , Cebidae/genética , Cercopithecidae/genética , Evolución Molecular , Antígenos HLA/química , Antígenos HLA-G , Antígenos de Histocompatibilidad Clase I/química , Humanos , Modelos Moleculares , Datos de Secuencia Molecular , Polimorfismo GenéticoRESUMEN
South American siskin radiation was studied by both mitochondrial cytochrome b (mt cyt b) DNA sequencing and homologous phenotypic characters; the latter were coded separately according to sex. Mixed phenetic and molecular (total evidence) dendrograms were constructed and the corresponding analyses suggest that speciation started in the South American siskin group with a north to south separation (Carduelis notata/C. barbata) along the Andean spine. A second split may have taken place around the Peruvian Andean mountains, corresponding to the present distribution pattern of C. olivacea. The most recent speciation events seem to have occurred in three sister species pairs: (i) C. xanthogastra/C. atrata, (ii) C. magellanica/C. yarrellii, (iii) C. cucullata/C. crassirostris. Accumulation of consistent characters in both morphological and molecular data at the basal nodes of the dendrograms indicate that speciation events occurred within a short period of time. Our data also suggest that speciation probably occurred by sexual selection through female mating choice in this radiation. Additionally, studies of variable amino acid residues in the mt cyt b molecule show that the three variable amino acids found are placed in the mitochondrial transmembrane region, which is also part of the hypervariable region in mammals. Each of the three amino acid changes occur in each of the three postulated evolutionary groups.
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Evolución Biológica , Variación Genética , Genética de Población , Filogenia , Pájaros Cantores/genética , Secuencia de Aminoácidos , Animales , Grupo Citocromo b/genética , ADN Mitocondrial/genética , Femenino , Geografía , Masculino , Datos de Secuencia Molecular , Fenotipo , Selección Genética , Análisis de Secuencia de ADN , Conducta Sexual Animal/fisiología , América del SurRESUMEN
Three new allelic forms of the HLA-G DNA sequence (HLA-G*II, HLA-G*III, and HLA-G*IV) have been identified. With the HLA-G*I sequence (previously designated HLA 6.0) as a reference, HLA-G*II shows a silent (G-->A) mutation at the third base of codon 57, HLA-G*III bears a non-synonymous (A-->T), but conservative, (Thr-->Ser) substitution at the first base of codon 31, and HLA-G*IV shows two silent substitutions: (A-->T) at the third base of codon 107 and (G-->A) at the third base of codon 57. A rapid method of singling out each allele on genomic DNA has been developed by using polymerase chain reaction amplification followed by restriction endonuclease treatment. Also, more or less strong linkage disequilibria has been found between most HLA-A alleles and either HLA-G*I or *II, both being the most prevalent alleles in the population, with a genotypic frequency of 0.55 and 0.38, respectively; HLA-G*III is very rare and HLA-G*IV has a genotypic frequency of 0.07. An evolutive classification of HLA-A alleles results according to their association with either HLA-G*I or HLA-G*II, which does not correlate with the classical serological cross-reacting groups classification. The finding of a strong and selective A/G linkage disequilibria with most HLA-A alleles, together with the existence of less frequent random A/G associations, may suggest that there exist in different haplotypes true and varied A/G genetic distances (and not a recombinational hotspot). It may be inferred from preliminary data that in primates HLA-A/G haplotypes bearing G*II may have appeared later than those bearing G*I.
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Alelos , Antígenos HLA/genética , Antígenos HLA-A/genética , Antígenos de Histocompatibilidad Clase I/genética , Desequilibrio de Ligamiento , Secuencia de Bases , Antígenos HLA-G , Humanos , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Polimorfismo Genético , Polimorfismo de Longitud del Fragmento de RestricciónRESUMEN
Immunomodulatory effects of different retinoids have been demonstrated, both in vivo and in vitro, in different cellular lineages including human and murine thymocytes, human lung fibroblasts, Langerhans' cells, tumoral cells and natural killer (NK) cells; however, any attempt to demonstrate the effect of retinoids on human peripheral blood mononuclear cells (PBMC) resulted in negative results. In the present work, it is shown that retinol and retinoic acid induce a marked increase of proliferation on human PBMC from 32 unrelated healthy individuals, which had previously been stimulated with anti-CD3 antibodies 48 hr before. Serum-free medium, specific retinoid concentration (10(-7) M) and a particular timing of retinol addition to the cultures (48 hr after CD3 stimulation) was necessary clearly to detect this retinol-enhancing effect. The increased proliferative response is specifically mediated via the clonotipic T-cell receptor-CD3 complex and correlates with the up-regulation of certain adhesion/activation markers on the T-lymphocyte surface: CD18, CD45RO and CD25; also Th1-type of cytokines (interleukin-2 and interferon-gamma) are found concordantly increased after retinoid costimulation, both measured by a direct protein measurement and by a specific mRNA increase. In addition, it is shown that the in vitro retinol costimulation is only present in immunodeficient patients who have no defect on CD3 molecules and activation pathway. The fact that retinol costimulate lymphocytes only via CD3 (and not via CD2 or CD28) and the lack of response enhancement in immunodeficients with impaired CD3 activation pathway indicates that retinoids may be used as therapeutic agents in immune system deficiencies that do not affect the clonotypic T-cell receptor.
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Complejo CD3/inmunología , Activación de Linfocitos/inmunología , Linfocitos T/inmunología , Vitamina A/inmunología , Técnicas de Cultivo de Célula , División Celular/inmunología , Medio de Cultivo Libre de Suero , Citocinas/biosíntesis , Humanos , Síndromes de Inmunodeficiencia/inmunología , Mitógenos/inmunologíaRESUMEN
Nijmegen breakage syndrome (NBS) is a genetic disorder characterized by immunodeficiency, microcephaly, and "bird-like" facies. NBS shares some clinical features with ataxia telangiectasia (AT), including increased sensitivity to ionizing radiation, increased spontaneous and induced chromosome fragility, and strong predisposition to lymphoid cancers. The mutated gene that results in NBS codes for a novel double-stranded DNA break repair protein, named nibrin. In the present work, a Spanish NBS patient was extensively characterized at the immunological and the molecular DNA levels. He showed low CD3(+)-cell numbers and an abnormal low CD4(+) naive cell/CD4(+) memory cell ratio, previously described in AT patients and also described in the present report in the NBS patient. The proliferative response of peripheral blood lymphocytes in vitro to mitogens is deficient in NBS patients, but the possible link among NBS mutations and the abnormal immune response is still unknown.
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Proteínas de Ciclo Celular/inmunología , Rotura Cromosómica/inmunología , Activación de Linfocitos/inmunología , Proteínas Nucleares , Linfocitos T/inmunología , Linfocitos B/inmunología , Linfocitos T CD4-Positivos/efectos de los fármacos , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/efectos de los fármacos , Linfocitos T CD8-positivos/inmunología , Proteínas de Ciclo Celular/genética , Niño , Humanos , Cariotipificación , Lectinas/metabolismo , Masculino , Mitógenos/farmacología , Mutación , Síndrome , Linfocitos T/efectos de los fármacos , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
Gamma interferon (IFN-gamma) and the cellular responses induced by it are essential for controlling mycobacterial infections. Most patients bearing an IFN-gamma receptor ligand-binding chain (IFN-gammaR1) deficiency present gross mutations that truncate the protein and prevent its expression, giving rise to severe mycobacterial infections and, frequently, a fatal outcome. In this report a new mutation that affects the IFN-gammaR1 ligand-binding domain in a Spanish patient with mycobacterial disseminated infection and multifocal osteomyelitis is characterized. The mutation generates an amino acid change that does not abrogate protein expression on the cellular surface but that severely impairs responses after the binding of IFN-gamma (CD64 and HLA class II induction and tumor necrosis factor alpha and interleukin-12 production). A patient's younger brother, who was also probably homozygous for the mutation, died from meningitis due to Mycobacterium bovis. These findings suggest that a point mutation may be fatal when it affects functionally important domains of the receptor and that the severity is not directly related to a lack of IFN-gamma receptor expression. Future research on these nontruncating mutations will make it possible to develop new therapeutical alternatives in this group of patients.
Asunto(s)
Interferón gamma/metabolismo , Infecciones por Mycobacterium no Tuberculosas/genética , Infección por Mycobacterium avium-intracellulare/genética , Mutación Puntual , Receptores de Interferón/genética , Inmunodeficiencia Combinada Grave/genética , Secuencia de Bases , Sitios de Unión , Línea Celular , Preescolar , Femenino , Humanos , Masculino , Datos de Secuencia Molecular , Infecciones por Mycobacterium no Tuberculosas/metabolismo , Complejo Mycobacterium avium , Infección por Mycobacterium avium-intracellulare/metabolismo , Micobacterias no Tuberculosas , Osteomielitis/genética , Osteomielitis/metabolismo , Linaje , Receptores de Interferón/metabolismo , Inmunodeficiencia Combinada Grave/complicaciones , Inmunodeficiencia Combinada Grave/metabolismo , Receptor de Interferón gammaRESUMEN
Leucocyte adhesion deficiency (LAD) is an autosomal-recessive genetic disease that is characterized clinically by severe bacterial infections and caused by mutations in the CD18 gene that codes for the beta2 integrin subunit. A patient with a severe LAD phenotype was studied and the molecular basis of the disease was identified as a single homozygous defect in a Herpes virus saimiri (HVS)-transformed T-cell line. The defect identified involves a deletion of 171 bp in the cDNA that encodes part of the proteic extracellular domain. This genetic abnormality was further studied at the genomic DNA level and found to consist of a deletion of 169 bp (from -37 of intron 4 to +132 of exon 5), which abolishes the normal splicing and results in the total skipping of exon 5. The 171-bp shortened 'in-frame' mRNA not only resulted in the absence of CD18 expression on the cell surface but also in its absence in the cytoplasm of HVS T-cell lines. Functionally, the LAD-derived HVS T-cell lines showed a severe, selective T-cell activation impairment in the CD2 (but not in the CD3) pathway. This defect was not reversible when exogenous interleukin-2 (IL-2) was added, suggesting that there is also a functional interaction of the lymphocyte function-associated antigen-1 (LFA-1) protein in the CD2 signal transduction pathway in human T cells, as has been previously reported in mice and in the human Papillon-Lefèvre syndrome. Thus, HVS transformation is not only a suitable model for T-cell immunodeficiency studies and characterization, but is also a good system for investigating the immune system in pathological conditions. It may also be used in the future in cellular models for in vitro gene-therapy trials.
Asunto(s)
Antígenos CD18/genética , Eliminación de Gen , Síndrome de Deficiencia de Adhesión del Leucocito/genética , Secuencia de Bases , Antígenos CD18/análisis , Antígenos CD2/inmunología , Línea Celular Transformada , Citoplasma/inmunología , Citometría de Flujo , Herpesvirus Saimiriino 2 , Homocigoto , Humanos , Lactante , Síndrome de Deficiencia de Adhesión del Leucocito/inmunología , Activación de Linfocitos , Antígeno-1 Asociado a Función de Linfocito/inmunología , Masculino , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Linfocitos T/inmunologíaRESUMEN
Three ataxia telangiectasia (AT) patients have been characterized immunologically and molecularly. Patient 1 presents two nondescribed splicing mutations which affect exons 15 and 21 of the ATM gene. The maternal defect consists of a G > A transition in the first nucleotide of the intron 21 donor splicing site which results in a complete deletion of exon 21. The paternal mutation consists of an A > C transversion in the intron 14 acceptor splicing site which produces a partial skipping of exon 15. Two abnormal alternative transcripts were found, respectively, 17 and 41 nucleotides shorter. Patient 2 presents a homozygous genomic deletion of 28 nucleotides in the last exon of the gene. This deletion changes the normal reading frame after residue 3003 of the protein and introduces a premature stop codon at residue 3008 that could originate a truncated ATM protein. Patient 3, a compound heterozygote, presents a defect which consists of a G > A transition in the first nucleotide of intron 62 donor splicing site which results in a complete deletion of exon 62. The results obtained during a three year period in the proliferation assays show an impaired PMA (phorbol myristate acetate) activation in specific T lymphocyte activation pathways (CD69, CD26, CD28, CD3, PHA, PWM and Con A mediated) but not in others (CD2, ionomycin, and Ig surface receptor). The possible link among specific ATM mutations and abnormal immune responses is unknown.