{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T21:43:32Z","timestamp":1791409412359,"version":"4.3.3"},"reference-count":39,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5446","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[1999,12,3]]},"abstract":"<jats:p>The signal transduction events that control the progenitor B cell (pro-B cell) to precursor B cell (pre-B cell) transition have not been well delineated. In evaluating patients with absent B cells, a male with a homozygous splice defect in the cytoplasmic adapter protein BLNK (B cell linker protein) was identified. Although this patient had normal numbers of pro-B cells, he had no pre-B cells or mature B cells, indicating that BLNK plays a critical role in orchestrating the pro-B cell to pre-B cell transition. The immune system and overall growth and development were otherwise normal in this patient, suggesting that BLNK function is highly specific.<\/jats:p>","DOI":"10.1126\/science.286.5446.1954","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:49:42Z","timestamp":1027763382000},"page":"1954-1957","source":"Crossref","is-referenced-by-count":291,"title":["An Essential Role for BLNK in Human B Cell Development"],"prefix":"10.1126","volume":"286","author":[{"given":"Yoshiyuki","family":"Minegishi","sequence":"first","affiliation":[{"name":"Department of Immunology,"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jurg","family":"Rohrer","sequence":"additional","affiliation":[{"name":"Department of Immunology,"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elaine","family":"Coustan-Smith","sequence":"additional","affiliation":[{"name":"Department of Hematology-Oncology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Howard M.","family":"Lederman","sequence":"additional","affiliation":[{"name":"Division of Pediatric Allergy and Immunology, Department of Pediatrics, Johns Hopkins Hospital, Baltimore, MD 21287, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rajita","family":"Pappu","sequence":"additional","affiliation":[{"name":"Center for Immunology, Division of Rheumatology, Departments of Medicine and Pathology, Howard Hughes Medical Institute, Washington University School of Medicine, St. Louis, MO 63110, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dario","family":"Campana","sequence":"additional","affiliation":[{"name":"Department of Hematology-Oncology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA."},{"name":"Department of Pediatrics, University of Tennessee, College of Medicine, Memphis, TN 38105, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew C.","family":"Chan","sequence":"additional","affiliation":[{"name":"Center for Immunology, Division of Rheumatology, Departments of Medicine and Pathology, Howard Hughes Medical Institute, Washington University School of Medicine, St. Louis, MO 63110, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mary Ellen","family":"Conley","sequence":"additional","affiliation":[{"name":"Department of Immunology,"},{"name":"Department of Pediatrics, University of Tennessee, College of Medicine, Memphis, TN 38105, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_1_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-7613(00)80591-9"},{"key":"e_1_3_1_3_2","doi-asserted-by":"crossref","unstructured":"J. Wienands et al. J. Exp. Med. 188 791 (1998);","DOI":"10.1084\/jem.188.4.791"},{"key":"e_1_3_1_3_3","doi-asserted-by":"crossref","first-page":"5804","DOI":"10.4049\/jimmunol.161.11.5804","volume":"161","author":"Goitsuka R.","year":"1998","unstructured":"Goitsuka R., et al., J. Immunol. 161, 5804 (1998).","journal-title":"J. Immunol."},{"key":"e_1_3_1_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-7613(00)80012-6"},{"key":"e_1_3_1_5_2","doi-asserted-by":"crossref","first-page":"1034","DOI":"10.1086\/301828","volume":"62","author":"Conley M. E.","year":"1998","unstructured":"Conley M. E., Mathias D., Treadaway J., Minegishi Y., Rohrer J., Am. J. Hum. Genet. 62, 1034 (1998).","journal-title":"Am. J. Hum. Genet."},{"key":"e_1_3_1_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(93)90667-F"},{"key":"e_1_3_1_6_3","doi-asserted-by":"publisher","DOI":"10.1038\/361226a0"},{"key":"e_1_3_1_7_2","doi-asserted-by":"crossref","first-page":"1486","DOI":"10.1056\/NEJM199611143352003","volume":"335","author":"Yel L.","year":"1996","unstructured":"Yel L., et al., N. Engl. J. Med. 335, 1486 (1996);","journal-title":"N. Engl. J. Med."},{"key":"e_1_3_1_7_3","doi-asserted-by":"publisher","DOI":"10.1084\/jem.187.1.71"},{"key":"e_1_3_1_8_2","doi-asserted-by":"crossref","first-page":"1115","DOI":"10.1172\/JCI7696","volume":"104","author":"Minegishi Y.","year":"1999","unstructured":"Minegishi Y., et al., J. Clin. Invest. 104, 1115 (1999).","journal-title":"J. Clin. Invest."},{"key":"e_1_3_1_9_2","unstructured":"The sequence flanking each exon of BLNK has been deposited in GenBank (accession numbers  through ). The primers used to amplify each exon are available upon request. The conditions used in SSCP are as described in work by M. E. Conley M. E. Fitch-Hilgenberg J. L. Cleveland O. Parolini and J. Rohrer [ Hum. Mol. Genet. 3 1751 (1994)]."},{"key":"e_1_3_1_10_2","unstructured":"The following primers were used to amplify BLNK exon 1 and the associated flanking sequence for both the SSCP analysis and cloning of this region of the gene: 5\u2032-GAACTGCTGACGTGACCA-3\u2032 (5\u2032 untranslated region of exon 1) and 5\u2032-CCCTAAAAGCTCAGTCCAC-3\u2032 (intron 1). The products of two independent PCR reactions were cloned and sequenced."},{"key":"e_1_3_1_11_2","unstructured":"DNA samples from five individuals demonstrated altered migration in comparison to the wild-type pattern. Sequencing showed a T to A substitution at the +113 position in intron 1 in three people a G to A substitution at the +121 position in intron 1 in one individual and an A to G substitution 5 base pairs upstream of the start codon."},{"key":"e_1_3_1_12_2","unstructured":"D. N. Cooper and M. Krawczak Human Gene Mutation (BIOS Oxford 1994) pp. 239\u2013260."},{"key":"e_1_3_1_13_2","unstructured":"The primers used to amplify BLNK cDNA were from the 5\u2032 untranslated region [shown in (9)] and 5\u2032-GTCGCTGTCAAAGTCATCGGA-3\u2032 from exon 4. The Btk-specific primers were from exons 6 and 8 of that gene. The primers used to amplify TdT \u03bb5 VDJ-\u03bc and glyceraldehyde phosphate dehydrogenase (GAPDH) were as reported in (7)."},{"key":"e_1_3_1_14_2","unstructured":"The father had 6% and the mother had 8% CD19 + B cells (normal is 5 to 22%). Serum IgG in the father was 1110 mg\/dl IgA was 337 mg\/dl and IgM was 93 mg\/dl. The mother's IgG was 1060 mg\/dl IgA was 115 mg\/dl and IgM was 127 mg\/dl."},{"key":"e_1_3_1_15_2","unstructured":"Bone marrow mononuclear cells were stained with antibody to CD19 conjugated to phycoerythrin (PE) (Dako Carpinteria CA) antibody to CD34 conjugated to peridinin chlorophyll protein (PerCP) (Becton-Dickinson San Jose CA) and polyclonal antibodies against human light chains conjugated to fluorescein isothiocyanate (FITC) (Southern Biotechnology Associates Birmingham AL). Antibodies to nuclear TdT (Supertechs Bethesda MD) and cytoplasmic \u03bc heavy chains (Southern Biotechnology Associates) conjugated to FITC and PE respectively were applied after cell permeabilization with OrthoPermeafix (Ortho Diagnostics Raritan NJ). Monoclonal antibody to BLNK (of IgG2a isotype) was generated against a glutathione S-transferase fusion protein encoding amino acids 4 through 205 as described in (1). This antibody was used in combination with an antibody to CD19 of IgM class (BLY3) (Research Diagnostics Flanders NJ) with fluorochrome-conjugated secondary antibodies specific for murine IgG and IgM. Antibody to BLNK was applied after cell permeabilization with OrthoPermeafix. Isotype-matched nonreactive antibodies were from Becton-Dickinson. Immunofluorescence staining was analyzed with a FACScan flow cytometer equipped with CellQuest software (Becton-Dickinson)."},{"key":"e_1_3_1_16_2","doi-asserted-by":"crossref","first-page":"7304","DOI":"10.1093\/emboj\/17.24.7304","volume":"17","author":"Zhang Y.","year":"1998","unstructured":"Zhang Y., Wienands J., Zurn C., Reth M., EMBO J. 17, 7304 (1998).","journal-title":"EMBO J."},{"key":"e_1_3_1_17_2","doi-asserted-by":"publisher","DOI":"10.1038\/381751a0"},{"key":"e_1_3_1_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/1074-7613(95)90114-0"},{"key":"e_1_3_1_18_3","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1016\/1074-7613(95)90115-9","volume":"3","author":"Kenner J. D.","year":"1995","unstructured":"Kenner J. 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L.","year":"1999","unstructured":"Law C. L., et al., J. Exp. Med. 189, 1243 (1999);","journal-title":"J. Exp. Med."},{"key":"e_1_3_1_22_9","doi-asserted-by":"crossref","first-page":"1068","DOI":"10.1002\/(SICI)1521-4141(199904)29:04<1068::AID-IMMU1068>3.0.CO;2-P","volume":"29","author":"Wunderlich L.","year":"1999","unstructured":"Wunderlich L., Farago A., Downward J., Buday L., Eur. J. Immunol. 29, 1068 (1999).","journal-title":"Eur. J. Immunol."},{"key":"e_1_3_1_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-7613(00)80032-1"},{"key":"e_1_3_1_24_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.281.5375.416"},{"key":"e_1_3_1_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)81422-1"},{"key":"e_1_3_1_26_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI5317"},{"key":"e_1_3_1_27_2","unstructured":"We thank M. D. Cooper for a helpful discussion E. Boylin for technical assistance and S. Saucier for administrative assistance. Supported by NIH grants AI25129 AI42787 CA71516 and CA58297; March of Dimes grant FY97-0384; the Assisi Foundation; National Cancer Institute CORE grant P30 CA21765; American Lebanese Syrian Associated Charities; the Human Frontiers Scientific Organization; and funds from the Federal Express Chair of Excellence. A.C.C. is a Pew Scholar in the Biomedical Sciences."}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.286.5446.1954","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,13]],"date-time":"2024-01-13T09:29:12Z","timestamp":1705138152000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.286.5446.1954"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1999,12,3]]},"references-count":39,"journal-issue":{"issue":"5446","published-print":{"date-parts":[[1999,12,3]]}},"alternative-id":["10.1126\/science.286.5446.1954"],"URL":"https:\/\/doi.org\/10.1126\/science.286.5446.1954","relation":{},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[1999,12,3]]}}}