PMID- 3111857
OWN - NLM
STAT- MEDLINE
DCOM- 19870910
LR  - 20051117
IS  - 0014-2980 (Print)
IS  - 0014-2980 (Linking)
VI  - 17
IP  - 7
DP  - 1987 Jul
TI  - Immunoglobulin heavy chain gene diversification in the long-term bone marrow 
      culture of normal mice and mice with severe combined immunodeficiency.
PG  - 1051-7
AB  - The change of immunoglobulin heavy (H) chain gene configuration during the 
      differentiation of B cells from their early precursors was investigated in 
      long-term cultures of bone marrow cells (LTBC). Hemopoietic stem cells are 
      maintained in LTBC described by Dexter et al. (J. Cell. Physiol. 1977. 91: 335; 
      LTBC-D), which supports the differentiation of myeloid lineage cells but not B 
      lineage cells. By simply shifting the culture condition to that devised by 
      Whitlock and Witte (Proc. Natl. Acad. Sci. USA 1982. 79: 3608; LTBC-B) to support 
      the development of B lineage cells, surface IgM-bearing (sIgM+) B cells became 
      detectable by the 2nd week after the shift and the number quickly increased 
      thereafter, while the number of polymorphonuclear cells and 
      granulocyte-macrophage colony-forming cell (CFU-c) decreased rapidly. H chain 
      gene configuration of the developing cells in the culture was examined by 
      Southern blot analysis of Eco RI-digested DNA with a JH probe. Whereas rearranged 
      JH gene configuration was not detectable in the DNA from LTBC-D cells, it first 
      appeared 2 weeks after the shift, and the level of the rearrangement rapidly 
      increased thereafter as the intensity of JH band of germ-line configuration 
      decreased. Almost all the cells in the culture had undergone H chain gene 
      rearrangement in both chromosomes by the 6th week after the shift. During 2 to 4 
      weeks after the shift, a cluster of bands spanning around 4.5-5.5 kb appeared 
      dominant among the rearranged configurations of JH gene and then it decreased in 
      intensity as the pattern of JH band became a more homogeneous smear. The 
      distribution of rearranged JH bands observed during 3-4 weeks after the shift was 
      strikingly similar to that observed in normal spleen B cells. By 
      semi-quantitative analysis of the intensity of JH and DSP2 bands remaining in 
      germ-line configuration, it was found that the loss of germ-line JH band was far 
      more rapid than that of germ-line DSP2 bands in the developing B cells in vitro. 
      This result is consistent with the conclusion obtained in B cell tumor lines that 
      D to J assembly occurred first and was followed by V to DJ assembly in this 
      culture system. Taken together, it is likely that the process of H chain gene 
      diversification in this culture system may represent the actual process during B 
      cell differentiation in vivo. Differentiative capacity of bone marrow stem cells 
      from mouse with severe combined immunodeficiency (SCID) was also analyzed in the 
      same culture system.(ABSTRACT TRUNCATED AT 400 WORDS)
FAU - Hirayoshi, K
AU  - Hirayoshi K
FAU - Nishikawa, S
AU  - Nishikawa S
FAU - Kina, T
AU  - Kina T
FAU - Hatanaka, M
AU  - Hatanaka M
FAU - Habu, S
AU  - Habu S
FAU - Nomura, T
AU  - Nomura T
FAU - Katsura, Y
AU  - Katsura Y
LA  - eng
PT  - Journal Article
PL  - Germany
TA  - Eur J Immunol
JT  - European journal of immunology
JID - 1273201
RN  - 0 (Antigens, Surface)
RN  - 0 (Culture Media)
RN  - 0 (Immunoglobulin Heavy Chains)
RN  - 0 (Immunoglobulin M)
RN  - 9004-70-0 (Collodion)
SB  - IM
MH  - Animals
MH  - Antigens, Surface/analysis
MH  - B-Lymphocytes/cytology/immunology
MH  - *Bone Marrow Cells
MH  - Cell Differentiation
MH  - Collodion
MH  - Colony-Forming Units Assay
MH  - Culture Media
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Genetic Code
MH  - Hematopoietic Stem Cells/cytology
MH  - Immunoglobulin Heavy Chains/*genetics
MH  - Immunoglobulin M/metabolism
MH  - Immunologic Deficiency Syndromes/*immunology
MH  - Mice
MH  - Mice, Inbred Strains/*immunology
MH  - Time Factors
EDAT- 1987/07/01 00:00
MHDA- 1987/07/01 00:01
CRDT- 1987/07/01 00:00
PHST- 1987/07/01 00:00 [pubmed]
PHST- 1987/07/01 00:01 [medline]
PHST- 1987/07/01 00:00 [entrez]
AID - 10.1002/eji.1830170723 [doi]
PST - ppublish
SO  - Eur J Immunol. 1987 Jul;17(7):1051-7. doi: 10.1002/eji.1830170723.