GM02471

GM02471 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:

No Data

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

White

Family Member

Relation to Proband

proband

Confirmation

Molecular characterization after cell line submission to CCR

Species

Homo sapiens

Common Name

Human

Remarks

Clinically affected; physical exam at birth was normal; subject was placed in protective isolation after birth and had good weight gain and no infections during the first 6 weeks; no thymic shadow seen on chest x-ray; perianal abscesses were intermittently present between one and ten months of age; normal development; normal kidneys but possible obstruction of left ureter; T cells were 44% and 46% at 3 and 9 days of age, respectively and B cells were 43% and 19%, respectively; leukopenia; immunoglobulin levels at 20 days of age were normal; deficient RBC & fibroblast ADA activity; residual ADA activity could be detected in mononuclear-cell/platelet preparations; less than 1.5% of ADA activity was seen in amniotic fluid cells; enzyme phenotypes: G6PD=B; 6PGD=A; peptidases A, C, & D=1; & PGM 1 & 3=1; donor subject is a compound heterozygote: one allele has a T>G transversion at nucleotide 911 in exon 10 of the ADA gene [911T>G] resulting in a substitution of arginine for leucine at codon 304 [Leu304Arg (L304R)] and a second allele has a C>T transition at nucleotide 466 of the ADA gene [466C>T] resulting in a substitution of cysteine for arginine at codon 156 [Arg156Cys (R156C)]; affected sister died at 9 months of age; mother is GM02472; same donor as GM02434 fibroblast.

Characterizations

IDENTIFICATION OF SPECIES OF ORIGIN

Species of Origin Confirmed by Nucleoside Phosphorylase, Glucose-6-Phosphate Dehydrogenase, and Lactate Dehydrogenase Isoenzyme Electrophoresis

MUTATION VERIFICATION

Adrian et al (Mol Cell Biol 4:1712-1717 1984) performed S1 endonuclease cleavage of ADA cDNA-mRNA hybrids to show that the ADA mRNA from this culture yielded fragments that were indistinguishable from ADA mRNA fragments from normal controls. The ADA cDNA utilized for these experiments represented the complete mRNA sequence with the possible exception of some of the 5 prime untranslated region. Valerio et al (EMBO J 5:113-119 1986) cloned and sequenced an ADA gene from this ADA-deficient cell line. Two independent cosmid clones were both shown to contain two point mutations resulting in amino acid substitutions in codons 80 (Lys to Arg) and 304 (Leu to Arg). Wildtype sequences were also detected at the same positions indicating compound heterozygosity. Studies with ADA expression clones mutagenized in vitro showed that the mutation at position 304 was responsible for ADA inactivation.

isoenzyme phenotype

AK1 isoenzyme phenotype is 1 (Valerio EMBO J 5:113 1986).

isoenzyme phenotype

Esterase D isoenzyme phenotype is 1 (Valerio EMBO J 5:113 1986).

isoenzyme phenotype

Alpha-L-Fucosidase 1 isoenzyme phenotype is 2-1 (Valerio EMBO J 5: 113 1986).

GENE MAPPING & DOSAGE STUDIES - Y CHROMOSOME

PCR analysis of DNA from this cell culture gave a positive result with a primer for Yq11, DYS227.

isoenzyme phenotype

Glyoxylase I isoenzyme phenotype is 2 (Valerio EMBO J 5:113 1986).

isoenzyme phenotype

PGD isoenzyme phenotype is A (Valerio EMBO J 5:113 1986).

isoenzyme phenotype

Phosphoglucomutase 1 isoenzyme phenotype is 1 (Valerio EMBO J 5:113 1986).

adenosine deaminase

According to the submitter, biochemical test results for this subject showed decreased enzyme activity. EC Number: 3.5.4.4; <1.5% activity.

Gene

ADA

Chromosomal Location

20q13.11

Allelic Variant 1

608958.0005; SEVERE COMBINED IMMUNODEFICIENCY, AUTOSOMAL RECESSIVE, T CELL-NEGATIVE, B CELL-NEGATIVE, NK CELL-NEGATIVE, DUE TO ADENOSINE DEAMINASE DEFICIENCY

Identified Mutation

LEU304ARG; In a patient with SCID due to ADA deficiency (102700) originally reported by Hirschhorn et al. (1975), Valerio et al. (1986) identified compound heterozygosity for 2 mutations in the ADA gene: one was a T-to-G transversion in exon 10, resulting in a leu304-to-arg (L304R) substitution. Functional expression studies showed that the L304R substitution resulted in ADA enzyme inactivation.

Gene

ADA

Chromosomal Location

20q13.11

Allelic Variant 2

608958.0018; SEVERE COMBINED IMMUNODEFICIENCY, AUTOSOMAL RECESSIVE, T CELL-NEGATIVE, B CELL-NEGATIVE, NK CELL-NEGATIVE, DUE TO ADENOSINE DEAMINASE DEFICIENCY

Identified Mutation

ARG156CYS; In 2 patients with SCID who were unusual for reportedly responding to the limited form of enzyme therapy provided by repeated partial exchange transfusions {Polmar et al. [N Engl J Med 295: 1337 (1976); Dyminski et al. [Clin Immunol Immunopathol 14: 307 (1979)]}, Hirschhorn [Hum Mutat 1: 166 (1992)] found two new missense mutations, arg156-to-cys and ser291-to-leu (102700.0019). The first of these was found in cell line GM02471 and represented a CGC-to-TGC transition at codon 156.

Phenotypic Data

Remarks

Publications

Lamontagne RJ, Soldan SS, Su C, Wiedmer A, Won KJ, Lu F, Goldman AR, Wickramasinghe J, Tang HY, Speicher DW, Showe L, Kossenkov AV, Lieberman PM, A multi-omics approach to Epstein-Barr virus immortalization of B-cells reveals EBNA1 chromatin pioneering activities targeting nucleotide metabolism PLoS pathogens17:e1009208 2020

PubMed ID: 33497421

Hirschhorn R, Identification of two new missense mutations (R156C and S291L) in two ADA- SCID patients unusual for response to therapy with partial exchange transfusions. Hum Mutat1:166-8 1992

PubMed ID: 1284479

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

Danton MJ, Coleman MS, Isolation of mutant adenosine deaminase by coformycin affinity chromatography. Anal Biochem159:233-9 1986

PubMed ID: 3492941

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

Hirschhorn R, Ellenbogen A, Martiniuk F, An approach to a selection system for adenosine-deaminase-positive (ADA+) cells and detection of rat ADA+ "revertants". J Cell Physiol123:277-82 1985

PubMed ID: 3872305

Adrian GS, Wiginton DA, Hutton JJ, Characterization of normal and mutant adenosine deaminase messenger RNAs by translation and hybridization to a cDNA probe. Hum Genet68:169-72 1984

PubMed ID: 6548726

Adrian GS, Wiginton DA, Hutton JJ, Structure of adenosine deaminase mRNAs from normal and adenosine deaminase-deficient human cell lines. Mol Cell Biol4:1712-7 1984

PubMed ID: 6208479

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

Daddona PE, Mitchell BS, Meuwissen HJ, Davidson BL, Wilson JM, Koller CA, Adenosine deaminase deficiency with normal immune function. An acidic enzyme mutation. J Clin Invest72:483-92 1983

PubMed ID: 6603477

Wiginton DA, Hutton JJ, Immunoreactive protein in adenosine deaminase deficient human lymphoblast cell lines. J Biol Chem257:3211-7 1982

PubMed ID: 6977542

Daddona PE, Kelley WN, Characteristics of an aminohydrolase distinct from adenosine deaminase in cultured human lymphoblasts. Biochim Biophys Acta658:280-90 1981

PubMed ID: 6972784

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

Polmar SH, Stern RC, Schwartz AL, Wetzler EM, Chase PA, Hirschhorn R, Enzyme replacement therapy for adenosine deaminase deficiency and severe combined immunodeficiency. N Engl J Med295:1337-43 1976

PubMed ID: 980079

Hirschhorn R, Beratis N, Rosen FS, Parkman R, Stern R, Polmar S, Adenosine-deaminase deficiency in a child diagnosed prenatally. Lancet1:73-75 1975

PubMed ID: 46025

Polmar SH, Wetzler EM, Stern RC, Hirschhorn R, Restoration of in-vitro lymphocyte responses with exogenous adenosine deaminase in a patient with severe combined immunodeficiency. Lancet2:743-6 1975

PubMed ID: 52774