RESUMEN
In areas without newborn screening for severe combined immunodeficiency (SCID), disease-defining infections may lead to diagnosis, and in some cases, may not be identified prior to the first year of life. We describe a female infant who presented with disseminated vaccine-acquired varicella (VZV) and vaccine-acquired rubella infections at 13 months of age. Immunological evaluations demonstrated neutropenia, isolated CD4 lymphocytopenia, the presence of CD8(+) T cells, poor lymphocyte proliferation, hypergammaglobulinaemia and poor specific antibody production to VZV infection and routine immunizations. A combination of whole exome sequencing and custom-designed chromosomal microarray with exon coverage of primary immunodeficiency genes detected compound heterozygous mutations (one single nucleotide variant and one intragenic copy number variant involving one exon) within the IL7R gene. Mosaicism for wild-type allele (20-30%) was detected in pretransplant blood and buccal DNA and maternal engraftment (5-10%) demonstrated in pretransplant blood DNA. This may be responsible for the patient's unusual immunological phenotype compared to classical interleukin (IL)-7Rα deficiency. Disseminated VZV was controlled with anti-viral and immune-based therapy, and umbilical cord blood stem cell transplantation was successful. Retrospectively performed T cell receptor excision circle (TREC) analyses completed on neonatal Guthrie cards identified absent TREC. This case emphasizes the danger of live viral vaccination in severe combined immunodeficiency (SCID) patients and the importance of newborn screening to identify patients prior to high-risk exposures. It also illustrates the value of aggressive pathogen identification and treatment, the influence newborn screening can have on morbidity and mortality and the significant impact of newer genomic diagnostic tools in identifying the underlying genetic aetiology for SCID patients.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Varicela/etiología , Linfopenia/etiología , Mutación , Receptores de Interleucina-7/genética , Rubéola (Sarampión Alemán)/etiología , Inmunodeficiencia Combinada Grave/genética , Vacunación/efectos adversos , Variaciones en el Número de Copia de ADN , Exoma , Femenino , Humanos , Lactante , Análisis de Secuencia por Matrices de Oligonucleótidos , Inmunodeficiencia Combinada Grave/inmunologíaRESUMEN
Synthetic substrates are often used to measure the activity of proteolytic enzymes. We have investigated the activities which cleave synthetic substrates such as alpha-N-benzyloxycarbonyl-Arg-Arg-beta-naphthylamide, for which the lysosomal proteinase cathepsin B has a high affinity, in sera from normal individuals, pregnant women and patients with breast cancer. As reported by other workers, activities against these substrates were elevated during pregnancy. Naphthylamine release, however, was shown to be the result of the combined action of two enzymes. The substrate is first cleaved by an endopeptidase to yield alpha-N-benzyloxycarbonyl-Arg and the aminopeptidase substrate Arg-beta-naphthylamide, which is then cleaved by serum aminopeptidases, particularly oxytocinase. A similar mechanism of cleavage was also found in the sera of breast cancer patients, where the endopeptidase catalyzing the first reaction was characterized as plasma kallikrein and the second reaction was carried out by serum leucine aminopeptidase. In no serum sample was there evidence for true cathepsin B activity.
Asunto(s)
Aminopeptidasas/sangre , Neoplasias de la Mama/sangre , Catepsinas/sangre , Endopeptidasas/sangre , Benzoilarginina-2-Naftilamida/análogos & derivados , Benzoilarginina-2-Naftilamida/metabolismo , Catepsina B , Cistinil Aminopeptidasa/sangre , Femenino , Humanos , Calicreínas/sangre , Leucil Aminopeptidasa/sangre , Embarazo , Especificidad por SustratoRESUMEN
The latent cysteine proteinase present in ascitic fluid of patients with neoplasia and released from ascites cells in culture has been partially purified and the enzyme after pepsin activation was shown to be immunologically related to the lysosomal proteinase, cathepsin B. The latent form was characterized as a single chain of Mr 40 000 as determined by SDS-polyacrylamide gel electrophoresis under reducing conditions followed by Western blotting and immune staining with an antiserum to human cathepsin B. Using the same techniques the enzyme after pepsin activation gave a single band of Mr 33 000. Analysis by isoelectric focusing showed that the latent enzyme before and after pepsin treatment is composed of several acidic isoenzymes. These findings suggest that this latent proteinase represents a precursor form of cathepsin B which is released extracellularly rather than being processed and directed to the lysosome.