De novo Generation of Cells within Human Nurse Macrophages and Consequences following HIV-1 Infection
Figure 3
Characteristics distinguishing CD4+ T-cells produced in primary macrophage cultures from those produced in EDTA/IL2-mac.
(A) As was the case in cultures of replated EDTA-mac treated with IL-2, the vast majority of T-cells produced in primary MDM cultures were also CD4+ T-cells. This is depicted here. However, in contrast to IL-2 treated cultures, these CD4+ T-cells were almost exclusively CD45RO+ and lacked CD45RA. Cells shown here were harvested on day 13 of culture and gated based on small size and CD3 expression (red arrows). The forward versus side scatter plots on the left demonstrate the relative numbers of large macrophages and lymphocytes within the nonadherent cell population. The numbers shown within the quadrants represent percentages from among the gated populations. (B) In contrast to those recovered from EDTA/IL2-mac, as shown here, the CD4+ T-cells produced in primary macrophage cultures did not express CD71, the transferrin receptor, indicating that they were resting lymphocytes. The CD71+ cells apparent are small CD4dim/CD14+ monocytoid cells, not lymphocytes. Analyses performed using Paint-a-Gate software. (C) Also in contrast to the CD4+ T-cells produced in IL-2-treated cultures, as shown here, 40–50% of these cells produced in primary MDM cultures expressed CD195, and 30–35% were dual positive for CD195 and CD184. Gates for the dot plots shown were established based on CD4bright expression and low side scatter, as indicated in the left panels (red arrows). Small, CD4dim cells were excluded from the gates because these were small monocytoid cells, not T-cells. The numbers shown within the quadrants represent percentages from among the gated populations. Analyses performed using CellQuest software. For (B) and (C), the letter D refers to the age of the culture in days at the time of harvest.