Primers used for <i>gag</i> and <i>pol</i> gene DNA identification in the peripheral blood of HIV-1 JRCSF infected humanized NSG mice.

<p>Primers used for <i>gag</i> and <i>pol</i> gene DNA identification in the peripheral blood of HIV-1 JRCSF infected humanized NSG mice.</p

Transplantation conditions and engraftment levels of human CD45⁺ cells after 22 weeks in various groups of NSG mice.

<p>Transplantation conditions and engraftment levels of human CD45⁺ cells after 22 weeks in various groups of NSG mice.</p

Sustained high-level of HIV dissemination and infection progression.

<p>(A) Humanized mice were i.v. injected with HIV-1 JRCSF and HIV-1 Bal at 10,000 TCID. Mice were bled at 20, 40, 60, 80, 100 and 120 days to detect plasma viral load, curve represent the viral load at different time point. Error bars (B) Sections of lymph node (a) and spleen (b) of humanized HIV infected mice were stained using anti-p24 antibody. Brown color show p24 specific staining. Lymph node of HIV-1 infected human (c) is shown as positive control and engrafted noninfected mouse spleen (d) is shown as negative control. (C) DNA was extracted from peripheral blood of HIV-1JRCSF-infected mice 9 week after infection. Determination of HIV-1 DNA copy was performed by PCR assay for <i>gag</i> and <i>pol</i> gene. 8E5 cell DNA was amplified as positive control (PC) along with humanized mice DNA (hm1, hm2, and hm3) (D, E) Percentage of CD4⁺ cells of T-cells following HIV-1 infection, each circle represent one mouse infected with HIV-1 JRCSF and each square represent one mouse infected with HIV-1 Bal means of CD4% of CD3⁺ cells at time point of infection are shown in solid line.</p

Coreceptor expression and lymphoid organ formation in humanized mice.

<p>(A) Representative FACS profile of human CCR5⁺CD4⁺ and CXCR4⁺CD4⁺ cells in spleen and lymph nodes, gate was set of human CD45 cells. Expression of CCR5 and CXCR4 were checked on CD14⁺ cells, gate was set on live human cell population. (B) Histology of lymphoid organs in CD34⁺ engrafted NSG mice. The lymphoid follicles mainly contained hCD45 cells. Spleen sections were stained with (a) anti-hCD45, (b) anti-hCD20 and (c) anti-hCD138. Lymph node sections were also stained with (d) anti-hCD45, (e) anti-hCD20 (f) anti-hCD3 (g,h) anti-hKi67 antibodies. Thymus section was stained with anti-CD3 antibody. (C) Proliferative responses of T cells were measured in nonstimulated cells, after stimulation with immobilized anti-CD3/anti-CD28 and anti-CD3+IL2 in spleen (white bars), lymph nodes (gray bars) and human PBMCs (black bars).</p

Engraftment levels of human cells in peripheral blood (PB), spleen, lymph nodes (LNs) and bone marrow (BM) of humanized NSG mice.

<p>Peripheral blood and organs were collected at 12 and 22 weeks after CD34⁺ cell transplantation in group 1 and group 7 mice. Percentage of human and murine CD45 cells was calculated on lymphoid gate. Percentage of human CD19⁺, CD3⁺, CD4⁺ and CD8⁺ cells were calculated out of human CD45⁺ cells.</p

Human cell differentiation in humanized NSG mice.

<p>(A) Absolute number of human cells in lymph nodes, bone marrow and spleens of group 1 and group 7 mice. (B) Mesenteric and brachial lymph nodes from male mice obtained after 24 weeks of CD34⁺ cell transplantation in group 7 mice. (C) Flow cytometric analysis of engraftment percentage of CD3⁺ T cells and CD19⁺ B cells at 12 and 22 weeks after transplantation of CD34⁺ cells in representative mice from group 1 and group 7. (D) Flow cytometric analysis of CD4⁺CD8⁺, CD4⁺ and CD8⁺ population in thymus of 22 weeks engrafted mice. (E) Flow cytometric analysis of CD45RA⁺ and CD45RO⁺ positive CD3 cells 22 weeks after engraftment in mice from group 7. (F) Representative profile CD11c⁺ cells in spleen and CD14⁺ population in bone marrow and blood of group 7 mice 22 weeks after CD34⁺ cell transplantation.</p

Engraftment potential of CD34⁺ hematopoietic cells.

<p>(A) Flow cytometric analysis of murine and human CD45⁺ cells in the peripheral blood of NSG mice of two different groups, representative profiles of the mice engraftment levels after 12 and 22 weeks after CD34⁺ cells transplantation. (B) Engraftment levels of human CD45⁺ cells in peripheral blood up to 306 days after transplantation in group 1 and group 7. (C) White blood cell counts at 12 and 22 weeks post-engraftment in group 7 mice along with human control.</p

HIV induced immune activation and expression levels of multiple activation markers on CD4⁺ and CD8⁺.

<p>Blood samples were obtained from mice before and 6 week after HIV-1 JRCSF infection. (A) Beta-2-microglobline levels were measured using ELISA before (Pre) and after HIV-infection (Post) (n = 4) and at same time intervals in engrafted but noninfected mice. (B) CD69 (C) HLA-DR (D) PD-1 and (E) CD27 (F) CCR5 levels were evaluated on CD4⁺ and CD8⁺ cells. (G) LPS levels in humanized mice 2 and 8 weeks post-HIV infection. Each circle and square represent one mouse at the indicated time point. Means are shown in solid lines. <i>P</i> values were determined by unpaired student’s t-tests. ** indicates a <i>P</i> value<0.001 and *** indicates a <i>P</i> value<0.0001.</p

Comparison of published busulfan-preconditioning protocols, kind and number of CD34⁺ cells transplanted and engraftment levels of human CD45⁺ cells in peripheral blood (PB), spleen and bone marrow (BM) of NSG mice.

<p>Comparison of published busulfan-preconditioning protocols, kind and number of CD34⁺ cells transplanted and engraftment levels of human CD45⁺ cells in peripheral blood (PB), spleen and bone marrow (BM) of NSG mice.</p

LLC metastasis growth in DUSP3^-/- mice is driven by hematopoietic cells.

<p>Tumours were established by injection (i.v.) of 10⁶ LLC-Luc+ cells to DUSP3^+/+ or DUSP3^-/- BM-transplanted irradiated DUSP3^+/+ mice. <b>(A)</b> Western blot analysis for DUSP3 expression from spleen lysates of transplanted mice (M = mouse). (<b>B</b>) Representative xenogen bioluminescence imaging results. <b>(C)</b> Quantitative imaging data (day 14). <b>(D)</b> Correlation between size of tumours (quantified as photons/second) and DUSP3 expression. <b>(E)</b> Comparison of lungs weights of DUSP3^+/+ or DUSP3^-/- BM-transplanted irradiated DUSP3^+/+ mice. <b>(F)</b> Hematoxylin-eosin staining of lung section from DUSP3^+/+ or DUSP3^-/- BM-transplanted irradiated DUSP3^+/+ mice. <b>(G)</b> Comparison of tumour areas from DUSP3^+/+ or DUSP3^-/- BM-transplanted irradiated DUSP3^+/+ mice. Student t-test was used for (<b>C</b>) and (<b>E</b>) and Mann-Whitney test was used for (<b>G</b>). *p < 0,05, **p < 0.01. 4 mice were used in each group and for each experiment. Data shown are representative of 2 different experiments.</p