Distinct mutations in two patients with leukocyte adhesion deficiency and their functional correlates.
Two patients with leukocyte adhesion deficiency (LAD), one with a moderate phenotype (patient 14) and one with a severe phenotype (patient 2) who had been shown to have a normal sized beta subunit protein precursor, were analyzed in an attempt to determine the molecular basis for their disease. RNase mapping located possible mutations to two distinct but adjacent regions of the beta subunit cDNA. Sequencing of patient-derived cDNA clones in this region revealed a C for T difference at amino acid 149 in patient 14 which resulted in the substitution of a leucine for a proline, and an A for G substitution at amino acid 169 in patient 2 which mutated a glycine to an arginine. The mutated amino acids are in a region of the cDNA that is highly conserved between the beta subunits of the integrin family and are identical in all known integrin beta subunits. Co-transfection of the beta subunit cDNA containing the patient 2 mutation with the wild-type alpha subunit of LFA-1 in a mammalian expression system resulted in no expression of LFA-1. In the case of the mutation in patient 14 there was markedly diminished expression of LFA-1 with loss of function and loss of the epitope for a number of anti-beta mAbs. Normal half-life of the mutant beta subunits, and previous demonstration of a lack of alpha/beta complex formation during biosynthesis in patient cells, suggest a defect in association with the alpha subunit. Association with beta is required for expression of the alpha subunit of LFA-1. Loss of functional expression with both of these beta subunit mutations suggests that they lie in a site critical for association with the alpha subunit.
Center for Blood Research, Harvard Medical School, Boston, Massachusetts 02115.
A J
AJ
Wardlaw
M L
ML
Hibbs
S A
SA
Stacker
T A
TA
Springer
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation / Q000172:deficiency / Q000235:genetics; D001483:Base Sequence; D018821:CD18 Antigens; D002448:Cell Adhesion; D002460:Cell Line; D004252:DNA Mutational Analysis; D005434:Flow Cytometry; D005785:Gene Conversion; D006801:Humans; D007153:Immunologic Deficiency Syndromes / Q000235:genetics* / Q000473:pathology; D018370:Leukocyte-Adhesion Deficiency Syndrome; D007962:Leukocytes / Q000473:pathology; D016169:Lymphocyte Function-Associated Antigen-1; D008969:Molecular Sequence Data; D009154:Mutation; D015345:Oligonucleotide Probes; D010641:Phenotype; D012313:RNA / Q000235:genetics; D016028:Receptors, Leukocyte-Adhesion / Q000235:genetics*; D014162:Transfection
D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D018821:CD18 Antigens; D016169:Lymphocyte Function-Associated Antigen-1; D015345:Oligonucleotide Probes; D016028:Receptors, Leukocyte-Adhesion; D012313:RNA