cd8 t cell positive selection kits  (Miltenyi Biotec)

Journal: Cancer immunology, immunotherapy : CII

Article Title: Absence of host NF-κΒ p50 induces murine glioblastoma tumor regression, increases survival, and decreases T cell induction of tumor-associated macrophage M2 polarization

doi: 10.1007/s00262-018-2184-2

Figure Lengend Snippet: Gliomas in p50−/− hosts have increased total and activated CD4 T cells and, to a lesser extent, increased activated CD8 T cells. a CD4 or CD8 cells were quantified as total number per tumor and the relative number per tumor bioluminescence 14 days after inoculation of GL261-Luc into WT or p50−/− mice (mean and SE from four determinations). b Representative Foxp3;CD25 FC plots within the CD3+CD4+ T cell subset (left) and quantification of Foxp3+CD25+ Tregs as a percent of CD4 cells (right, mean and SE from four determinations) are shown. c Representative CD4;IFNγ and CD8;IFNγ FC plots, within the CD3+ T cell gate, are shown 4 hrs after exposure to vehicle or PMA/ionomycin (left). CD4+IFNγ+ or CD8+IFNγ+ cells were quantified as the relative number per bioluminescence unit (right, mean and SE from four determinations). d Representative CD3;IFNγ, CD3;Granzyme B (GrzB), and CD3;TNFα FC plots, within CD4+ or CD8+ gates, are shown three days after stimulation of isolated CD3+ tumor cells with Dynabeads containing CD3 and CD28 antibodies (left). IFNγ+, GrzB+, and TNFα+ cells were quantified as percent of CD4+ or CD8+ cells (right, mean and SE from five determinations).

Article Snippet: Naïve splenic T cell analysis To obtain naive CD4 T cells, spleens were passed through a cell strainer, subjected to red cell lysis, and selected using a CD8 positive selection kit (Miltenyi).

Techniques: Isolation

Journal: Cancer immunology, immunotherapy : CII

Article Title: Absence of host NF-κΒ p50 induces murine glioblastoma tumor regression, increases survival, and decreases T cell induction of tumor-associated macrophage M2 polarization

doi: 10.1007/s00262-018-2184-2

Figure Lengend Snippet: Survival advantage of p50−/− mice is eliminated by depletion of CD4 T cells and partially eliminated by CD8 depletion, and combined CD4 and CD8 depletion reverses WT tumor myeloid M2 polarization. a WT and p50−/− mice received 200 μg CD4 or CD8 antibody (Ab) on days −7, −5, and −3, followed by GL261-Luc cell inoculation on d0, as diagrammed (left). Representative CD4;CD8 FC plots demonstrating efficient depletion of CD4 or CD8 T cells from the peripheral blood one day prior to GL261 inoculation (right). b Survival curves for glioma-bearing WT mice, p50−/− mice, or p50−/− mice pre-treated with CD4 or CD8 Ab. The number of mice in each group and Log Rank p-values comparing the control and Ab-treated p50- /- groups are shown. The untreated WT and p50−/− mice in this experiment are a subset of those presented in Fig. 1 and were inoculated concurrently with those exposed to the T cell-depleting Abs. c WT and p50−/− mice received both CD4 and CD8 antibody (Ab), or no Ab, on days −7, - 5, and −3, followed by GL261-Luc cell inoculation on d0 and an additional CD4/CD8 Ab injection on d8. CDllb+ cells isolated on day 14 were subjected to qRT-PCR analysis for indicated mRNAs (mean and SE from three determinations).

Article Snippet: Naïve splenic T cell analysis To obtain naive CD4 T cells, spleens were passed through a cell strainer, subjected to red cell lysis, and selected using a CD8 positive selection kit (Miltenyi).

Techniques: Control, Injection, Isolation, Quantitative RT-PCR