Investigation of a novel approach to scoring Giemsa-stained malaria-infected thin blood films-3

E same slide via manual scoring by a different researcher (correlation = .96). . Semi-automated scores of two images derived from different areas of the same slide, analysed by the same researcher, plotted against each other (correlation = .99).<p><b>Copyright information:</b></p><p>Taken from "Investigation of a novel approach to scoring Giemsa-stained malaria-infected thin blood films"</p><p>http://www.malariajournal.com/content/7/1/62</p><p>Malaria Journal 2008;7():62-62.</p><p>Published online 21 Apr 2008</p><p>PMCID:PMC2358916.</p><p></p

Investigation of a novel approach to scoring Giemsa-stained malaria-infected thin blood films-2

Of 16000×, to confirm that the software has accurately identified them as two separate cells. . The user has zoomed in on a cell that is ambiguous re infection status under standard magnification, and confirmed that it is infected.<p><b>Copyright information:</b></p><p>Taken from "Investigation of a novel approach to scoring Giemsa-stained malaria-infected thin blood films"</p><p>http://www.malariajournal.com/content/7/1/62</p><p>Malaria Journal 2008;7():62-62.</p><p>Published online 21 Apr 2008</p><p>PMCID:PMC2358916.</p><p></p

Investigation of a novel approach to scoring Giemsa-stained malaria-infected thin blood films-1

automatic total cell count, user correction thereof and parasitaemia scoring; blue crosses indicate where the user has added cells to the total cell-count, and green dots indicate where the user has subtracted cells. Yellow circles indicate cells that the user has deemed infected.<p><b>Copyright information:</b></p><p>Taken from "Investigation of a novel approach to scoring Giemsa-stained malaria-infected thin blood films"</p><p>http://www.malariajournal.com/content/7/1/62</p><p>Malaria Journal 2008;7():62-62.</p><p>Published online 21 Apr 2008</p><p>PMCID:PMC2358916.</p><p></p

The 'interphase' after automatic identification of individual cells and gridding of these ranked on various features potentially discriminating infected and uninfected cells

<p><b>Copyright information:</b></p><p>Taken from "Investigation of a novel approach to scoring Giemsa-stained malaria-infected thin blood films"</p><p>http://www.malariajournal.com/content/7/1/62</p><p>Malaria Journal 2008;7():62-62.</p><p>Published online 21 Apr 2008</p><p>PMCID:PMC2358916.</p><p></p

Investigation of a novel approach to scoring Giemsa-stained malaria-infected thin blood films-0

N arrows indicate unifected RBCs, and red arrows indicate infected RBCs. In this example the user has opted not to discriminate between the different stages of infected RBC. . A scored image of a human RBC culture experimentally infected with , captured at 600× magnification. In this example the user has chosen to specifically determine the number of trophozoites present (blue arrows), and infected cells at any other stage of development have simply been scored as 'infected RBC' (red arrows).<p><b>Copyright information:</b></p><p>Taken from "Investigation of a novel approach to scoring Giemsa-stained malaria-infected thin blood films"</p><p>http://www.malariajournal.com/content/7/1/62</p><p>Malaria Journal 2008;7():62-62.</p><p>Published online 21 Apr 2008</p><p>PMCID:PMC2358916.</p><p></p

Investigation of a novel approach to scoring Giemsa-stained malaria-infected thin blood films-5

N arrows indicate unifected RBCs, and red arrows indicate infected RBCs. In this example the user has opted not to discriminate between the different stages of infected RBC. . A scored image of a human RBC culture experimentally infected with , captured at 600× magnification. In this example the user has chosen to specifically determine the number of trophozoites present (blue arrows), and infected cells at any other stage of development have simply been scored as 'infected RBC' (red arrows).<p><b>Copyright information:</b></p><p>Taken from "Investigation of a novel approach to scoring Giemsa-stained malaria-infected thin blood films"</p><p>http://www.malariajournal.com/content/7/1/62</p><p>Malaria Journal 2008;7():62-62.</p><p>Published online 21 Apr 2008</p><p>PMCID:PMC2358916.</p><p></p

Foxp3 induction is MHC class II-dependent.

<p>MHC class-II dependency was examined using 5 µg/ml neutralizing anti-HLA DR or isotype control mAb. Cells were analysed for CD25 and Foxp3 expression on CD4 T cells on day 6 (A). Culture supernatants were sampled on day 2 (IL-2) or day 6 (IL-10) for cytokine analysis by ELISA (B). Fold changes in the proportion of Foxp3^hi and Foxp3^int cells (black bars) were calculated by normalizing for each donor the percentage of each of the two cell types of CD4⁺ T cells or cytokine levels on the respective values obtained for PBMCs cultured with iRBCs in the absence of any antibody (white bars). Experiments were set up with a 2∶1 iRBC∶PBMC ratio.</p

Supplementary Figure 5 from Reducing TNF Receptor 2⁺ Regulatory T Cells via the Combined Action of Azacitidine and the HDAC Inhibitor, Panobinostat for Clinical Benefit in Acute Myeloid Leukemia Patients

PDF file - 1448K, The proportion of TNFR2- and TNFR2+ effector T cells between healthy and AML patients.</p

Expansion of iRBC-induced Foxp3^hi and Foxp3^int CD4⁺CD25^hi T cells from CD4⁺CD25⁻ and CD25⁺ T cells.

<p>PBMC were stained with CFSE prior to set up of iRBC∶PBMC co-cultures and analysed on day 6 (A). Data for dividing and non-dividing CD4⁺ T cells were analysed as percentage of total Foxp3^hi and Foxp3^int CD4⁺ T cells from 7 donors (Mean+/−SEM) (B). Representative FACS plots for 1 out of 3 donors show the development of Foxp3 expression and CFSE dilution over a time course of 6 days of iRBC co-culture (C) To determine the origin of Foxp3^hi and Foxp3^int CD4⁺CD25^hi T cells, PBMC were depleted from CD25⁺ cells prior to set up with iRBCs. A representative dot plot demonstrating depletion efficiency of the CD25⁺Foxp3⁺ nTreg population is shown (D). Cells were then analysed over a time course (day 3 to 6) for the kinetics of CD25 and Foxp3 induction by flow cytometry (E). A representative dot plot for 1 out of 15 donors showing the generation of CD25^hiFoxp3^hi and CD25^hiFoxp3^int on day 6 is shown. Percentages of both populations are expressed within total CD4 T cells (fine print) or within the CD25^hi population (bold print) (F). Due to a general decrease in the induction of total CD25^hiCD4⁺ cells from CD25 depleted versus whole PBMCs, Foxp3^hi or Foxp3^int induction was analysed as percentage within the CD25^hi CD4 population. Data are represented as box-and-whisker-plots, with boxes extending from the 25^th to the 75^th percentile and horizontal lines representing the median, while whiskers extend to the lowest and highest data point (G). Experiments were set up with an iRBC∶PBMC ratio of 2∶1.</p

Monocytes and CD4 T cells are the main cellular source of TGFβ1 and IL-10 in iRBC∶PBMC co-cultures.

<p>On day 6 of iRBC∶PBMC co-culture (1∶5 ratio), individual cell types were separated by MACS and FACS to subsequently isolate RNA from purified populations. Relative mRNA levels (normalized on 18SrRNA) for TGFβ1 and IL-10 were determined and normalized for each donor on respective levels in total unseparated PBMC (A). The proportion of individual cell populations expressed as percent of total PBMC was determined by flow cytometry (B). Relative mRNA levels for IL-10 and TGFβ1 for each cell population were corrected for their proportion in whole PBMC by multiplying relative mRNA levels with the percentage of each population in whole PBMCs for each donor (C). Data (Mean+/−SEM) for 3 donors are shown.</p