Figure 5.

HSP70i has prolonged effects on peripheral leukocyte composition. Splenocytes obtained from Pmel-1 mice nine months after gene gun vaccination with HSP70i or empty vector DNA were stained to discriminate leukocyte subpopulations, and staining was quantified by flow cytometry. (A) Three distinct populations of cells were observed after gating for CD11b and CD11c cells in the non-lymphocyte population, with high (hi), low (lo) or intermediate (int) levels of expression. Macrophages are observed in the R1 population as CD11b^int/hiCD11c^lo cells, while dendritic cells are amongst the R2 and R3 populations as CD11b^intCD11c^int and CD11b^loCD11c^hi cells, respectively. (B) Quantification of CD11b and CD11c-expressing cells determined differential expression of these markers among leukocytes after vaccination with HSP70i encoding or empty vector plasmids. The macrophage population (CD11b^int/hiCD11c^lo) shown in R1 displays decreased quantitative differences in response to HSP70i. By contrast, the inflammatory dendritic cell population shown in R2 (CD11b^intCD11c^int) shows an opposite trend. These results demonstrate the differential effects HSP70i on monocyte derivative leukocyte populations. Student’s t-test *P < 0.05, n=3.