GM01715

GM01715 LCL from B-Lymphocyte

Description:

SEVERE COMBINED IMMUNODEFICIENCY, AUTOSOMAL RECESSIVE, T CELL-NEGATIVE, B CELL-NEGATIVE, NK CELL-NEGATIVE, DUE TO ADENOSINE DEAMINASE DEFICIENCY
ADENOSINE DEAMINASE; ADA

Affected:

Yes

Sex:

Male

Age:

3 YR (At Sampling)

Overview

Repository

NIGMS Human Genetic Cell Repository

Subcollection

Heritable Diseases

Class

Disorders of Nucleotide and Nucleic Acid Metabolism

Biopsy Source

Peripheral vein

Cell Type

B-Lymphocyte

Tissue Type

Blood

Transformant

Epstein-Barr Virus

Sample Source

LCL from B-Lymphocyte

Race

Black/African American

Family Member

Relation to Proband

proband

Confirmation

Clinical summary/Case history

Species

Homo sapiens

Common Name

Human

Remarks

Clinically affected; failure to thrive; multiple pneumonias and oral/skin moniliasis; long-term hospitalization starting at age 26 months: immunologic studies showed variable but persistent lymphopenia, low numbers of T lymphocytes, relatively increased percentage of B lymphocytes, bone marrow aspirate with few lymphocytes or plasma cells, lymph node biopsy revealed a "hypoplastic node with follicles but largely absent T-dependent areas and sinus histiocytosis", quantitative immunoglobins remained normal; height and weight increased to 25th-50th percentile by age 6 years but subject remained lymphopenic; between ages 11-14 years the absolute lymphocyte count was normal with a normal percentage of T cells; no medical problems at age 20 years (no theapy from age 13-20 years); HLA type A2,30,B7,39; enzyme phenotypes: 6PGD=A, G6PD=A, Peptidases A,C,&D=1, Acid A-glucosidase=1, Neutral A-glucosidase C=3, & GLO-1=2; 46,XY; ADA activity shortly after diagnosis was 1% of normal in erythrocytes and 18% of normal in mixed mononuclear cells; ADA in both cultured lymphoid and fibroblast cell lines established at diagnosis showed deficiency of ADA with <1% of normal ADA activity; a lymphoid cell line established at age 16 years exhibited >50% of normal ADA activity; studies performed on the lymphoid and fibroblast cell lines established at the time of diagnosis found the donor subject to be a compound heterozygote: one allele has a G>A transition at nucleotide 302 in exon 4 of the ADA gene [302G>A] resulting in a substitution of glutamine for arginine at codon 101 [Arg101Gln(R101Q)] and a second allele has a splice-donor-site mutation in IVS1(+1GT>CT) resulting in an unstable mRNA; the B cell line expressing ADA activity (established at age 16 years) contained the missense Arg101Gln mutation but lacked the splice-site mutation; donor subject determined to have somatic mosaicism; same donor as GM02445 fibroblast.

Characterizations

IDENTIFICATION OF SPECIES OF ORIGIN

Species of Origin Confirmed by Chromosome Analysis

MUTATION VERIFICATION

Bonthron et al (J Clin Invest 76:894-897 1985) reported the cloning and sequence analysis of full-length cDNAs from this ADA-deficient cell line. Four apparently identical though independently isolated clones were sequenced. In each a G to A mutation at base number 302 of the coding sequence was found which predicts a glutamine residue to be substituted for arginine in the ADA protein at codon 101. The authors believed that only one of the defective alleles in this cell line had been sequenced.

adenosine deaminase

According to the submitter, biochemical test results for this subject showed decreased enzyme activity. EC Number: 3.5.4.4; 1% activity.

Gene

ADA

Chromosomal Location

20q13.11

Allelic Variant 1

608958.0003; SEVERE COMBINED IMMUNODEFICIENCY, AUTOSOMAL RECESSIVE, T CELL-NEGATIVE, B CELL-NEGATIVE, NK CELL-NEGATIVE, DUE TO ADENOSINE DEAMINASE DEFICIENCY

Identified Mutation

ARG101GLN; In cell line GM01715 from an immunodeficient patient, Bonthron et al. [J Clin Invest 76: 894(1985)] found a point mutation in codon 101 (CGG to CAG) of ADA; this change predicts an amino acid change from arginine to glutamine. The mutation was apparently responsible for loss of function in the gene because the predicted primary structure of the enzyme was otherwise entirely normal. The demonstration of 2 different mutations in codon 101 leading to ADA deficiency indicates that this amino acid position is critical for stability and/or activity of the enzyme protein.

Gene

ADA

Chromosomal Location

20q13.11

Allelic Variant 2

608958.0024; SEVERE COMBINED IMMUNODEFICIENCY, AUTOSOMAL RECESSIVE, T CELL-NEGATIVE, B CELL-NEGATIVE, NK CELL-NEGATIVE, DUE TO ADENOSINE DEAMINASE DEFICIENCY

Identified Mutation

IVS1DS, G>C, +1; Hirschhorn et al. [Ann Hum Genet 58: 1 (1994)] found that fibroblast and B-cell lines established at the time of diagnosis of ADA deficiency (GM02445 and GM01715) were heteroallelic for a newly identified splice site mutation (+1 GT-to-CT transversion) at the donor splice site in IVS1 and for a previously described arg101-to-gln missense mutation in exon 4 (102700.0003). As described earlier, by the time the patient was 16 years of age, the mutation had disappeared from the B cells but not from the fibroblasts and the patient had undergone spontaneous recovery from ADA deficiency.

Phenotypic Data

Remarks

Publications

Hirschhorn R, Yang DR, Israni A, Huie ML, Ownby DR, Somatic mosaicism for a newly identified splice-site mutation in a patient with adenosine deaminase-deficient immunodeficiency and spontaneous clinical recovery. Am J Hum Genet55(1):59-68 1994

PubMed ID: 8023852

Hirschhorn R, Ellenbogen A, Tzall SHirschhorn, Five missense mutations at the adenosine deaminase locus (ADA) detected by altered restriction fragments and their frequency in ADA--patients with severe combined immunodeficiency (ADA-SCID). Am J Hum Genet42:201-7 1992

PubMed ID: 1346349

Hirschhorn R, Chakravarti V, Puck J, Douglas SD, Homozygosity for a newly identified missense mutation in a patient with very severe combined immunodeficiency due to adenosine deaminase deficiency (ADA-SCID). Am J Hum Genet49:878-85 1991

PubMed ID: 1680289

Tzall S, Ellenbogen A, Eng F, Hirschhorn R, Identification and characterization of nine RFLPs at the adenosine deaminase (ADA) locus. Am J Hum Genet44:864-75 1989

PubMed ID: 2567118

Akeson AL, Wiginton DA, Dusing MR, States JC, Hutton JJ, Mutant human adenosine deaminase alleles and their expression by transfection into fibroblasts. J Biol Chem263:16291-6 1988

PubMed ID: 3182793

Akeson AL, Wiginton DA, States JC, Perme CM, Dusing MR, Hutton JJ, Mutations in the human adenosine deaminase gene that affect protein structure and RNA splicing. Proc Natl Acad Sci U S A84:5947-51 1987

PubMed ID: 3475710

Berkvens TM, Gerritsen EJ, Oldenburg M, Breukel C, Wijnen JT, van Ormondt H, Vossen JM, van der Eb AJ, Meera Khan P, Severe combined immune deficiency due to a homozygous 3.2-kb deletion spanning the promoter and first exon of the adenosine deaminase gene. Nucleic Acids Res15:9365-78 1987

PubMed ID: 3684597

Valerio D, Dekker BM, Duyvesteyn MG, van der Voorn L, Berkvens TM, van Ormondt H, van der Eb AJ, One adenosine deaminase allele in a patient with severe combined immunodeficiency contains a point mutation abolishing enzyme activity. EMBO J5:113-9 1986

PubMed ID: 3007108

Bonthron DT, Markham AF, Ginsburg D, Orkin SH, Identification of a point mutation in the adenosine deaminase gene responsible for immunodeficiency. J Clin Invest76:894-7 1985

PubMed ID: 3839802

Daddona PE, Shewach DS, Kelley WN, Argos P, Markham AF, Orkin SH, Human adenosine deaminase. cDNA and complete primary amino acid sequence. J Biol Chem259:12101-6 1984

PubMed ID: 6090454

Valerio D, Duyvesteyn MG, van Ormondt H, Meera Khan P, van der Eb AJ, Adenosine deaminase (ADA) deficiency in cells derived from humans with severe combined immunodeficiency is due to an aberration of the ADA protein. Nucleic Acids Res12:1015-24 1984

PubMed ID: 6198631

Hirschhorn R, Roegner V, Jenkins T, Seaman C, Piomelli S, Borkowsky W, Erythrocyte adenosine deaminase deficiency without immunodeficiency. Evidence for an unstable mutant enzyme. J Clin Invest64:1130-9 1979

PubMed ID: 479373

Wolf, Syndrome of cellular immunodeficiency with immunoglobulins associated with adenosine deaminase deficiency. Pediatr Res10:394 (1976):1130-9 1976

PubMed ID: 479373