Figure 7.
External milieu drives MoMac behavior. Total peritoneal lavage cells were isolated from donor mice at 18 hours post–zymosan injection and adoptively transferred (AT) into peritonea of time-matched recipient mice. Recovery and phenotype of AT MoMacs were assessed 24 hours after adoptive transfer (42 hours post–zymosan injection). (A) Phenotype of MoMacs from WT (blue) or CGD (red) donor mice AT into WT (left) or CGD (right) recipient animals, overlaid onto MoMacs of the recipient animal (gray). (B) Pie charts show the percentage of total AT MoMacs with the indicated phenotype. Reported statistics represent results of analysis with reference to the transfer of WT PL cells into WT recipient animals. (C) Recovery in peritoneal lavage of AT MoMacs as a percentage of the input MoMac number, from either WT (blue) or CGD (red) recipient animals. (D) Immunofluorescence analysis of omentum (OME, top) and diaphragm (DIA, bottom) tissues stained as indicated. Green arrows indicate the location of AT CGD MoMacs in WT omentum and associated with CGD diaphragm. Original magnification, 10×. (E) Recovery of AT MoMacs in omentum (left) or diaphragm (right) digests analyzed by flow cytometry. Note difference in the y-axis. (F and G) WT and CGD mixed bone marrow chimeras (supplemental Figure 7 describes experimental design) were administered i.p. zymosan and harvested at 96 hours after zymosan injection. Peritoneal lavage fluid was assessed by flow cytometry, and diaphragm and omentum were assessed by immunofluorescence. (F) Number of MoMacs, subsetted based on Ly6C and MHCII expression, in WT (blue) and CGD (red and purple) chimeras. MoMacs within the CGD chimeras were further separated based on Nox2 expression, either Nox2-sufficient (purple) or Nox2-deficient (red). (G) Immunofluorescence staining of omentum (left) and diaphragm (right) from WT (top) and CGD (bottom) mixed chimeras. All images were 10× original magnification; they were acquired by using an Olympus VS200 and analyzed by using VS200 ASW software. All data represent 8 to 10 mice per group over 2 to 3 independent experiments. Error bars indicate standard error of the mean. *P < .05, **P < .01, ***P < .001. DAPI, 4′,6-diamidino-2-phenylindole; DIC, differential interference contrast.

External milieu drives MoMac behavior. Total peritoneal lavage cells were isolated from donor mice at 18 hours post–zymosan injection and adoptively transferred (AT) into peritonea of time-matched recipient mice. Recovery and phenotype of AT MoMacs were assessed 24 hours after adoptive transfer (42 hours post–zymosan injection). (A) Phenotype of MoMacs from WT (blue) or CGD (red) donor mice AT into WT (left) or CGD (right) recipient animals, overlaid onto MoMacs of the recipient animal (gray). (B) Pie charts show the percentage of total AT MoMacs with the indicated phenotype. Reported statistics represent results of analysis with reference to the transfer of WT PL cells into WT recipient animals. (C) Recovery in peritoneal lavage of AT MoMacs as a percentage of the input MoMac number, from either WT (blue) or CGD (red) recipient animals. (D) Immunofluorescence analysis of omentum (OME, top) and diaphragm (DIA, bottom) tissues stained as indicated. Green arrows indicate the location of AT CGD MoMacs in WT omentum and associated with CGD diaphragm. Original magnification, 10×. (E) Recovery of AT MoMacs in omentum (left) or diaphragm (right) digests analyzed by flow cytometry. Note difference in the y-axis. (F and G) WT and CGD mixed bone marrow chimeras (supplemental Figure 7 describes experimental design) were administered i.p. zymosan and harvested at 96 hours after zymosan injection. Peritoneal lavage fluid was assessed by flow cytometry, and diaphragm and omentum were assessed by immunofluorescence. (F) Number of MoMacs, subsetted based on Ly6C and MHCII expression, in WT (blue) and CGD (red and purple) chimeras. MoMacs within the CGD chimeras were further separated based on Nox2 expression, either Nox2-sufficient (purple) or Nox2-deficient (red). (G) Immunofluorescence staining of omentum (left) and diaphragm (right) from WT (top) and CGD (bottom) mixed chimeras. All images were 10× original magnification; they were acquired by using an Olympus VS200 and analyzed by using VS200 ASW software. All data represent 8 to 10 mice per group over 2 to 3 independent experiments. Error bars indicate standard error of the mean. *P < .05, **P < .01, ***P < .001. DAPI, 4′,6-diamidino-2-phenylindole; DIC, differential interference contrast.

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