Requirement for B cell linker protein (BLNK) in B cell development

Science

Volumn 286, Issue 5446, 1999, Pages 1949-1954

  Pappu, Rajita ^a   Cheng, Alec M ^a   Li, Bin ^a   Gong, Qian ^a   Chiu, Chris ^a   Griffin, Nancy ^a   White, Mike ^a   Sleckman, Barry P ^a   Chan, Andrew C ^a  

^a WASHINGTON UNIVERSITY SCHOOL OF MEDICINE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ADAPTOR PROTEIN; IMMUNOGLOBULIN G; IMMUNOGLOBULIN M; LYMPHOCYTE RECEPTOR; MITOGEN ACTIVATED PROTEIN KINASE; PROTEIN TYROSINE KINASE;

ANIMAL CELL; ARTICLE; B LYMPHOCYTE; B LYMPHOCYTE DIFFERENTIATION; BONE MARROW CELL; ENZYME ACTIVATION; ENZYME LOCALIZATION; MOUSE; NONHUMAN; PRIORITY JOURNAL; PROTEIN PHOSPHORYLATION; SIGNAL TRANSDUCTION;

ADAPTOR PROTEINS, SIGNAL TRANSDUCING; AGING; ANIMALS; B-LYMPHOCYTE SUBSETS; B-LYMPHOCYTES; BONE MARROW CELLS; CARRIER PROTEINS; CELL COUNT; CELL DIFFERENTIATION; CELL SEPARATION; CELL SIZE; FLOW CYTOMETRY; GENE TARGETING; HEMATOPOIETIC STEM CELLS; IMMUNOGLOBULIN M; LEUKOPOIESIS; LYMPHOID TISSUE; MICE; MICE, INBRED C57BL; PHOSPHOPROTEINS; RECEPTORS, ANTIGEN, B-CELL; SECOND MESSENGER SYSTEMS; SIGNAL TRANSDUCTION;

EID: 0033521085     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.286.5446.1949     Document Type: Article

References (51)

1. R. Benschop and J. Cambier, Curr. Opin. Immunol. 11, 143 (1999); K. Campbell, Curr. Opin. Immunol. 11, 256 (1999); T. Kurosaki, Annu. Rev. Immunol. 17, 555 (1999).
2. R. Benschop and J. Cambier, Curr. Opin. Immunol. 11, 143 (1999); K. Campbell, Curr. Opin. Immunol. 11, 256 (1999); T. Kurosaki, Annu. Rev. Immunol. 17, 555 (1999).
3. R. Benschop and J. Cambier, Curr. Opin. Immunol. 11, 143 (1999); K. Campbell, Curr. Opin. Immunol. 11, 256 (1999); T. Kurosaki, Annu. Rev. Immunol. 17, 555 (1999).
4. J. Healy and C. Goodnow, Annu. Rev. Immunol. 16, 645 (1998).
5. C. Fu, C. Turck, T. Kurosaki, A. Chan, Immunity 9, 93 (1998).
6. L. Gangi-Peterson et al., Mol. Immunol. 33, 55 (1998); S. Hashimoto et al., Blood 94, 2357 (1999).
7. L. Gangi-Peterson et al., Mol. Immunol. 33, 55 (1998); S. Hashimoto et al., Blood 94, 2357 (1999).
8. R. Goitsuka et al., J. Immunol. 161, 5804 (1998); J. Wienands et al., J. Exp. Med. 188, 791 (1998).
9. R. Goitsuka et al., J. Immunol. 161, 5804 (1998); J. Wienands et al., J. Exp. Med. 188, 791 (1998).
10. M. Ishiai et al., Immunity 10, 117 (1999).
11. 6) were prepared with the CytoFix/CytoPerm Kit (PharMingen) according to manufacturer's recommendations. After permeabilization, cells were stained with 2 μg of an antiserum to BLNK (3) in buffer containing 50% fetal calf serum (FCS) at 4°C for 30 min. Cells were washed twice with 1 ml of wash buffer and stained with 1 μg of fluorescein isothiocyanate-conjugated goat anti-rabbit IgG (heavy and light chains) (Jackson Research Labs) for 30 min at 4°C. Cells were washed twice with 1 ml of wash buffer, resuspended in 0.4 ml of cold phosphate-buffered saline, and prepared for FACS analysis.
12. R. Hardy, C. Carmarck, S. Shinton, J. Kemp, K. Hayakawa, J. Exp. Med. 173, 1213 (1991).
13. A. M. Cheng et al., Nature 378, 303 (1995); M. Turner et al., Nature 378, 298 (1995); M. Turner et al., J. Exp. Med 186, 2013 (1997).
14. A. M. Cheng et al., Nature 378, 303 (1995); M. Turner et al., Nature 378, 298 (1995); M. Turner et al., J. Exp. Med 186, 2013 (1997).
15. A. M. Cheng et al., Nature 378, 303 (1995); M. Turner et al., Nature 378, 298 (1995); M. Turner et al., J. Exp. Med 186, 2013 (1997).
16. J. L. Clements et al., Science 281, 416 (1998); V. Pivniouk et al., Cell 94, 229 (1998); J. Clements et al., J. Clin. Invest. 103, 19 (1999).
17. J. L. Clements et al., Science 281, 416 (1998); V. Pivniouk et al., Cell 94, 229 (1998); J. Clements et al., J. Clin. Invest. 103, 19 (1999).
18. J. L. Clements et al., Science 281, 416 (1998); V. Pivniouk et al., Cell 94, 229 (1998); J. Clements et al., J. Clin. Invest. 103, 19 (1999).
19. J. Chen, R. Lansford, V. Stewart, F. Young, F. Alt, Proc. Natl. Acad. Sci. U.S.A. 90, 4528 (1993).
20. +/- splenocytes or bone marrow-derived cells; this may be due to transcriptional silencing of GFP by the PGK-neo cassette. The targeting construct was linearized and electroporated into 129/ SVJ ES cells at 250 mV and 500 μF with a Gene Pulser II (Bio-Rad, Hercules, CA). Cells were selected in neomycin (0.2 mg/ml) and clones were expanded after 7 days of selection. Fifteen hundred neomycin-resistant clones were screened, of which three represented correct recombinants.
21. DNA was harvested from mouse tails and digested with Bam HI. A 500-base pair (bp) fragment 6 kb upstream of exon 1 was used as the 5′ probe, and a downstream 300-bp Xba I-Bam HI fragment served as the 3′ probe.
22. R. Pappu and A. Chan, unpublished results.
23. +/- ES cells, individual clones were subcloned for one round before injection into blastocysts.
24. J. Ledbetter, J. Goding, T. Tsu, L. Herzenberg, Immunogenetics 8, 347 (1979).
25. 6) were lysed in 10 mM tris (pH 8.0), 150 mM NaCl, and 1% NP-40 buffer containing protease and phosphatase inhibitors (3). Cell debris was clarified at 14,000g for 10 min at 4°C. The supernatant fraction was then analyzed by SDS-polyacrylamide gel electrophoresis and immunoblotting with antisera to BLNK (3) or actin (14). The antiserum to BLNK was raised against amino acids 1 through 204 and recognizes amino acids 61 through 204.
26. +/- mice.
27. Supplementary information is available at Science Online at www.sciencemag.org/feature/data/ 1045303.shl
28. Statistical analysis is presented as mean ± SD; n represents sample size and P values are derived from comparisons of independent sample tests.
29. 6 cells were examined. All antibodies were purchased from PharMingen. Data were collected with a FACS Calibur (Becton Dickinson, San Jose, CA) and analyzed with Cell Quest Analysis software. All data were collected from live cells within the lymphocyte gate as defined by forward and side scatter values.
30. F. Loder et al., J. Exp. Med. 190, 75 (1999).
31. Serum Ig levels were quantitated with the use of an enzyme-linked immunosorbent assay (ELISA) from Southern Biotechnology according to the manufacturer's recommendations.
32. D. Kitamura et al., Cell 69, 823 (1992).
33. J. G. Cyster et al., Nature 381, 325 (1996).
34. + B cells every 30 s by FACS analysis.
35. + B cells.
36. L. A. Herzenberg et al., Immunol. Rev. 93, 81 (1986); A. B. Kantor and L. A. Herzenberg, Annu. Rev. Immunol. 11, 501 (1993).
37. L. A. Herzenberg et al., Immunol. Rev. 93, 81 (1986); A. B. Kantor and L. A. Herzenberg, Annu. Rev. Immunol. 11, 501 (1993).
38. A. B. Kantor, A. M. Stall, S. Adams, L. A. Herzenberg, L. A. Herzenberg, Proc. Natl. Acad. Sci. U.S.A. 89, 3320 (1992).
39. H. von Boehmer et al., Curr. Opin. Immunol. 11, 135 (1999).
40. Y. Minegishi et al., Science 286, 1954 (1999).
41. S. Tsukada et al., Cell 72, 279 (1993); D. Vetrie et al., Nature 361, 226 (1993); W. N. Khan et al., Immunity 3, 283 (1995); J. D. Kerner et al., Immunity 3, 301 (1995); R. W. Hendriks et al., EMBO J. 13, 4862 (1996); Y. Minegishi et al., J. Exp. Med. 187, 71 (1998).
42. S. Tsukada et al., Cell 72, 279 (1993); D. Vetrie et al., Nature 361, 226 (1993); W. N. Khan et al., Immunity 3, 283 (1995); J. D. Kerner et al., Immunity 3, 301 (1995); R. W. Hendriks et al., EMBO J. 13, 4862 (1996); Y. Minegishi et al., J. Exp. Med. 187, 71 (1998).
43. S. Tsukada et al., Cell 72, 279 (1993); D. Vetrie et al., Nature 361, 226 (1993); W. N. Khan et al., Immunity 3, 283 (1995); J. D. Kerner et al., Immunity 3, 301 (1995); R. W. Hendriks et al., EMBO J. 13, 4862 (1996); Y. Minegishi et al., J. Exp. Med. 187, 71 (1998).
44. S. Tsukada et al., Cell 72, 279 (1993); D. Vetrie et al., Nature 361, 226 (1993); W. N. Khan et al., Immunity 3, 283 (1995); J. D. Kerner et al., Immunity 3, 301 (1995); R. W. Hendriks et al., EMBO J. 13, 4862 (1996); Y. Minegishi et al., J. Exp. Med. 187, 71 (1998).
45. S. Tsukada et al., Cell 72, 279 (1993); D. Vetrie et al., Nature 361, 226 (1993); W. N. Khan et al., Immunity 3, 283 (1995); J. D. Kerner et al., Immunity 3, 301 (1995); R. W. Hendriks et al., EMBO J. 13, 4862 (1996); Y. Minegishi et al., J. Exp. Med. 187, 71 (1998).
46. S. Tsukada et al., Cell 72, 279 (1993); D. Vetrie et al., Nature 361, 226 (1993); W. N. Khan et al., Immunity 3, 283 (1995); J. D. Kerner et al., Immunity 3, 301 (1995); R. W. Hendriks et al., EMBO J. 13, 4862 (1996); Y. Minegishi et al., J. Exp. Med. 187, 71 (1998).
47. D. H. Chu et al., J. Immunol. 163, 2610 (1999); A. C. Chan et al., Science 264, 1599 (1994); M. E. Elder et al., Science 264, 1596 (1994); E. Arpaia et al., Cell 76, 947 (1994).
48. D. H. Chu et al., J. Immunol. 163, 2610 (1999); A. C. Chan et al., Science 264, 1599 (1994); M. E. Elder et al., Science 264, 1596 (1994); E. Arpaia et al., Cell 76, 947 (1994).
49. D. H. Chu et al., J. Immunol. 163, 2610 (1999); A. C. Chan et al., Science 264, 1599 (1994); M. E. Elder et al., Science 264, 1596 (1994); E. Arpaia et al., Cell 76, 947 (1994).
50. D. H. Chu et al., J. Immunol. 163, 2610 (1999); A. C. Chan et al., Science 264, 1599 (1994); M. E. Elder et al., Science 264, 1596 (1994); E. Arpaia et al., Cell 76, 947 (1994).
51. Supported in part by grants from the Human Frontiers Program in Biomedical Sciences (to A.C.C.), the Pew Scholars Program in Biomedical Sciences (to A.C.C.), NIH grants AI42787 (to A.C.C) and CA71516 (to A.C.C.), and the Burroughs Wellcome Fund (to B.P.S.).