Flow cytometric analysis of recombinant murine GM-CSF (rmuGM-CSF) induced changes in the distribution of specific cell populations in vivo
✍ Scribed by Kenneth J. Pennline; Frances Pellerito; Manuel DaFonseca; Peggy Monahan; Marvin I. Siegel; Sidney R. Smith
- Publisher
- John Wiley and Sons
- Year
- 1990
- Tongue
- English
- Weight
- 748 KB
- Volume
- 11
- Category
- Article
- ISSN
- 0196-4763
No coin nor oath required. For personal study only.
✦ Synopsis
The changes in the distribution of granulocytes, monocytes, and lymphocytes in various tissue compartments following subcutaneous (SC) administration of recombinant murine GM-CSF (rmuGM-CSF) in vivo was determined by flow cytometry in time course studies. Balblc mice were given single, daily SC injections of 1 or 4 pg of rmuGM-CSF for 10 days. Flow cytometric analysis was performed on bone marrow (BMC), peritoneal exudate (PEC), and peripheral blood (PBC) cell preparations from mice treated for 1,3, and 10 days. Dual fluorescence was employed to gate on leukocytes (T200+) and analyze for Ig+, Thy 1.2+, MAC+, and 8C5+ (granulocytes) cells. The analyses indicated that SC-rmuGM-CSF increased the percentage of 8C5+ cells in PBC after 1 day of treatment. However, significant changes in the cell composition of PEC and BMC were not observed until day 10 of treatment and included increases in 8C5+ cells and the myeloid cell population, respectively. Side scatter analysis (cell density) of PBC and PEC indicated that the percentage of the granulocytic cell population increased significantly in rmuGM-CSF treated mice. The changes observed in PEC and BMC appeared to be doserelated whereas those observed in PBC were not. These data clearly demonstrate the utility of flow cytometric analyses for detecting selective effects of cytokines on cell populations that are involved in host defense mechanisms.
Key terms: Flow cytometry, light scattering, granulocytes, mouse, peripheral blood, bone marrow, peritoneal exudate 1 pg rmuGM-CSF 16 t 2** 25 2 2 8 2 1 43 + 3 16 i 2** 4.3 i 0.5** 1.2 2 0.1** 7.0 i 1.5** 2.3 i 0.4** aDuring these time course studies animals received single daily subcutaneous injections of 1 or 4 pg rmuGM-CSF in 0.2 ml bPercentages are based on T200+ cells. 'Results represent the mean 2 standard error for four individual experiments. Values from individual experiments were 4 pg rmuGM-CSF 4 2 1 30 + 3 5 2 2 50 2 6 4 i 2 1 pg rmuGM-CSF 4 t O 32 2 2 6 2 2 49 2 3 3 2 1 Vehicle 4 2 1 32 + 3 5 + 1 56 i 4 3 t l 4 pg rmuGM-CSF 8 2 1** 26 & 4 5 t 2 52 2 10 I 2* 1 pg rmuGM-CSF 6 2 1 27 2 2 6 t 2 52 2 8 8 2 3 10 Vehicle 5 + 0 33 2 4 11 2 3 41 2 4 6 i 1 of sterile saline. obtained from PEC pooled from three mice in each group. *P 5 0.05. **P 5 0.01.