Requirement for invariant chain in B cell maturation and function

Science

Volumn 274, Issue 5284, 1996, Pages 106-108

  Shachar, Idit ^a   Flavell, Richard A ^a  

^a YALE UNIVERSITY SCHOOL OF MEDICINE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CELL SURFACE MARKER; CHAPERONE; MAJOR HISTOCOMPATIBILITY ANTIGEN CLASS 2;

ANTIGEN EXPRESSION; ANTIGEN PRESENTATION; ARTICLE; B LYMPHOCYTE; CELL FUNCTION; CELL MATURATION; IMMUNE RESPONSE; PRIORITY JOURNAL; PROTEIN FOLDING; PROTEIN TRANSPORT;

ANIMALS; ANTIGENS; ANTIGENS, DIFFERENTIATION, B-LYMPHOCYTE; B-LYMPHOCYTES; BONE MARROW CELLS; CELLS, CULTURED; HISTOCOMPATIBILITY ANTIGENS CLASS II; IMMUNIZATION; IMMUNOGLOBULIN D; IMMUNOGLOBULIN M; LYMPHOCYTE ACTIVATION; MICE; MICE, KNOCKOUT; MICE, TRANSGENIC; RECEPTORS, IGE;

EID: 0029742201     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.274.5284.106     Document Type: Article

References (24)

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7. I. Shachar, E. A. Elliott, B. Chasnoff, I. S. Grewal, R. A. Flavell, Immunity 3, 373 (1995).
8. p41lo, 9 ± 1.
9. p41lo mice were harvested and were stained by FACS for B220 expression 9 days after KLH injection (19, 20).
10. + cells are due to an intrinsic property of these B cells.
11. 3H]thymidine incorporation for 12 hours.
12. M. F. Naujokas et al. Immunity 3, 359 (1995).
13. p41lo mice were double-stained with anti-B220 and anti-IgM, anti-HSA, or anti-CD23 (19). The markers expressed on the B cells from these mice were almost identical to those expressed on splenocytes from li mice.
14. Purified B lymphocytes from wild-type or li mice were cultured in the presence of different concentrations of LPS. Cells were then double-stained with anti-B220 and anti-IgM, anti-HSA, or anti-CD23. B cells from li mice expressed immature markers after LPS stimulation.
15. C. H. Chang, S. Guerder, S. C. Hong, W. van Ewijk, R. A. Flavell, Immunity 4, 167 (1996).
16. J. P. van Meerwijk and R. N. Germain, Semin, Immunol. 6, 231 (1994).
17. S. H. Chan, C. Benoist, D. Mathis, ibid., p. 241.
18. Control, li ,CD4 ,or IL-4 mice were triple-stained with anti-B220, anti-IgM, or anti-CD23 (19). The FACS analysis showed that B cells from the CD4 or IL-4 mice expressed almost identical amounts of the markers as did B cells from wild-type mice.
19. b; H 129.1.9 for CD4, 53-6.7 for CD8, M1/69 for heatstable antigen (HSA), S7 for CD43, B3B4 for CD23, 14.8 for CD45RA (B220), R6-60.2 for IgM, and AMS 9.1 for IgD. Bone marrow, lymph nodes, and spleen cell suspensions were prepared in Bruff's medium, and the spleen erythrocytes were lysed by hypotonic shock. Cells were resuspended in cold phosphate-buffered saline (PBS) supplemented with 1% FBS. Staining was performed in the same buffer.
20. Mice were injected in the hind footpads with 100 μg of KLH emulsified in complete Freund's adjuvant. Draining lymph nodes were collected 9 days later.
21. 16-CGG was with 100 μg of Alum-precipitated NP-CGG in 0.1 ml of 0.85% NaCl.
22. Splenocytes from wild-type, li ,or CIITA mice were incubated in digitonin (50 μg/ml). The pellet was then lysed in 0.5% Triton X-100, 300 mM NaCl, 50 mM tris (pH 7.4), 1 mM polymethylsulfonyl fluoride, leupeptin (10 μg/ml), aprotinin (10 μg/ml), pepstatin (10 μg/ml), chymostatin (10 μg/ml), and 20 mM N-ethyl-maleimide. Nuclei and debris were eliminated by centrifugation. Lysates were separated on 12% (w/v) SDS-polyacrylamide gel electrophoresis. The proteins were transferred onto nitrocellulose, which was blocked and then incubated with IN1 (monoclonal antibody to the li cytoplasmic tail) followed by horseradish peroxidase-conjugated goat antibody to rat IgG.
23. 6 cells were injected intravenously.
24. Supported by the Howard Hughes Medical Institute (R.A.F.) and the Irvington Institute (I.S.). We thank T. Geiger, I. S. Grewal, C.-H. Chang, P. Cresswell, M. Shlomchik, and L. Cohn.