Changes in mouse MHC expression during the cell cycle of Con A-treated spleen cells.
This study was directed at correlating the expression of class I MHC determinants with different segments of the cell cycle by using dual laser flow microfluorometry to measure levels of both DNA and cell surface H-2Kd or H-2Dd determinants for single cells. Con A-treated mouse spleen cells were identified as being in the G2/M or G0/G1 phases of the cell cycle on the basis of propidium iodide or Hoechst 33342 dye bound to DNA. Monoclonal anti-H-2 antibodies, indirectly fluoresceinated with goat anti-mouse IgG, were used to detect MHC determinants. The average level of both Kd and Dd determinants expressed by G2/M cells was about 1.6-fold higher than that expressed by G0/G1 cells. These observations indicate that the average-size G0/G1 and G2/M cells have the same apparent surface density of Kd and Dd determinants, insofar as we estimate that these cells differ in surface area by a factor of about 1.5. We also analyzed the expression of Kd and Dd determinants by measuring how they changed as a function of the intensity of forward light scatter from cells. For both G2/M and G0/G1 cells, changes in light scatter intensity were associated with parallel changes in levels of Dd and Kd determinants, indicating a common mechanism(s) that controls their cell surface expression.
C P
CP
Muller
D A
DA
Stephany
J R
JR
Wunderlich
D000818:Animals; D002453:Cell Cycle; D003208:Concanavalin A / Q000494:pharmacology*; D005260:Female; D005434:Flow Cytometry; D006183:H-2 Antigens / Q000032:analysis*; D007399:Interphase; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000166:cytology* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008938:Mitosis; D011419:Propidium; D013154:Spleen / Q000166:cytology; D013194:Staining and Labeling
D006183:H-2 Antigens; C024755:H-2K(K) antigen; D003208:Concanavalin A; D011419:Propidium