peripheral blood mononuclear cells pbmcs  (Axis-Shield Diagnostics)

Journal: Virology Journal

Article Title: Hepatitis B virus (HBV)-specific T-cell responses to recombinant HBV core protein in patients with normal liver function and co-infected with chronic HBV and human immunodeficiency virus 1 (HIV-1)

doi: 10.1186/1743-422X-10-232

Figure Lengend Snippet: Comparison of HBV-specific cell responses in the presence or absence of HBeAg. (A) C protein-specific T-cell responses elicited in ELISPOT assay; (B) Cytokine IFN-Y responses in cultured supernatants of PBMCs stimulated by C protein. HBeAg(−): HBeAg-negative subjects; HBeAg(+): HBeAg-positive subjects; HIV+: HIV co-infected; HIV-: not co-infected with HIV.

Article Snippet: Human interferon (IFN)-γ ELISPOT assay Peripheral whole blood was obtained from all subjects and peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation with Ficoll Lymphoprep (Axis –Shield PoC AS, Oslo, Norway).

Techniques: Enzyme-linked Immunospot, Cell Culture, Infection

Journal: Oncotarget

Article Title: Impact of lenalidomide maintenance on the immune environment of multiple myeloma patients with low tumor burden after autologous stem cell transplantation

doi: 10.18632/oncotarget.24944

Figure Lengend Snippet: Lenalidomide maintenance therapy does not enhance MM-specific CD4 + T-cell responses Analysis of T-cell responses specific for the MAGE-C1 peptide SQSSPVSSFPSSTSS. ( A ) CD137 expression on CD4 + and CD8 + T cells from patient#02 at the pre LEN timepoint upon in vitro restimulation with the MAGE-C1 peptide. ( B ) PBMCs obtained at the pre LEN and the LEN timepoint were examined by IFN-γ ELISPOT to measure the T-cell response to the MAGE-C1 peptide. The bar graphs show the mean number of spots per million cells of four replicate wells and the error bars represent the standard deviation. A two-way ANOVA with Sidak’s multiple comparisons test was performed to evaluate statistical significant differences, ns: not significant, ** p ≤ 0.01, *** p ≤ 0.001 and **** p ≤ 0.0001.

Article Snippet: Peripheral blood mononuclear cells (PBMCs) were isolated from leukapheresis (pre-LEN timepoint) and whole blood (LEN timepoint) samples through lymphoprep gradient-centrifugation (Axis-Shield) following the manufacturer’s instructions before being cryopreserved.

Techniques: Expressing, In Vitro, Enzyme-linked Immunospot, Standard Deviation

Journal: Oncotarget

Article Title: Impact of lenalidomide maintenance on the immune environment of multiple myeloma patients with low tumor burden after autologous stem cell transplantation

doi: 10.18632/oncotarget.24944

Figure Lengend Snippet: Lenalidomide maintenance therapy results in an improved CD8 + T cell functionality, which is hampered by non-CD8 + T cells present in the periphery ( A ) PBMCs obtained at the pre LEN and the LEN timepoint were examined by IFN-γ ELISPOT to measure the CD8 + T-cell response to viral recall epitopes derived for CMV, EBV and influenza (CEF) ( n = 6). The number of spot forming units (SFU) as well as the activity in each well was determined. Bar graphs show the mean number of spots per million PBMCs or the mean activity of four replicate wells and the error bars show the standard deviation. Spot numbers and activity measured upon culture in the absence of peptide were subtracted. ( B – E ) CD8 + T cells purified from PBMC samples obtained at the pre LEN and LEN timepoint were examined by IFN-γ/IL-2/TNF-α FLUOROSPOT to characterize the CD8 + T-cell response to the CEF peptide pool. (B–C) Bar graphs show the mean number of spots per million CD8 + T cells of five replicate wells and the error bars show the standard deviation. Spot numbers and activity measured upon culture in the absence of peptide were subtracted. ( D ) Raw data showing three-color FLUOROSPOT results. One representative well is shown for each patient and timepoint. Cytokine-specific spots are shown in green (IFN-γ), red (TNF-α), blue (IL-2) or overlays of these colors. ( E ) The pie charts indicate the portion of mono- and polyfunctional CEF-specific CD8 + T cells. A two-way ANOVA with Sidak’s multiple comparisons test was performed to detect differences between the pre LEN and the LEN timepoint, ns: not significant, * p

Article Snippet: Peripheral blood mononuclear cells (PBMCs) were isolated from leukapheresis (pre-LEN timepoint) and whole blood (LEN timepoint) samples through lymphoprep gradient-centrifugation (Axis-Shield) following the manufacturer’s instructions before being cryopreserved.

Techniques: Enzyme-linked Immunospot, Derivative Assay, Activity Assay, Standard Deviation, Purification

Journal: Experimental and Therapeutic Medicine

Article Title: Interleukin-1 receptor antagonist expression is inversely associated with outcomes of hepatitis B-related acute-on-chronic liver failure

doi: 10.3892/etm.2017.4361

Figure Lengend Snippet: Inhibition of cytokine secretion in LPS-stimulated PBMCs by rhIL-1ra. LPS, lipopolysaccharide; PBMCs, peripheral blood mononuclear cells; rh, human recombinant; IL, interleukin; ra, receptor antagonist; low, LPS + 125 ng/ml rhIL-1ra; middle, LPS + 250 ng/ml rhIL-1ra; high, LPS + 500 ng/ml rhIL-1ra; IFN, interferon; TNF, tumor necrosis factor.

Article Snippet: Incubation of peripheral blood mononuclear cells (PBMCs) with IL-1ra in vitro PBMCs were isolated from the heparinized blood of 31 patients with ACLF via centrifugation at 800 × g at 20°C for 20 min using Ficoll Lymphoprep (Axis-Shield Diagnostics Ltd., Dundee, UK).

Techniques: Inhibition, Recombinant

Journal: Mucosal immunology

Article Title: Novel role for IL-22 in protection during chronic Mycobacterium tuberculosis HN878 infection

doi: 10.1038/mi.2017.15

Figure Lengend Snippet: Infection with Mtb HN878 triggers increased IL-22 production through a TLR2-dependent pathway Lung cells isolated from B6, Il1r-/- and Tlr2-/- mice were infected with either Mtb H37Rv (Rv) or Mtb HN878 (HN) at MOI of 0.1 for 7 days, and the protein levels of ( a ) IL-1β and ( b ) IL-22 were assessed in culture supernatants by ELISA and luminex assay, respectively. Lung cells isolated from B6 mice were treated with cell wall extracted from Mtb H37Rv (Rv) or Mtb HN878 (HN), (10μg/ml each). Protein levels of ( c ) IL-1β and ( d ) IL-22 were assessed by ELISA and luminex assay respectively. PBMCs were isolated from ATB patients and were stimulated with Mtb H37Rv (Rv) or Mtb HN878 (HN) cell wall extracts (20 μg/ml each) and the protein levels of ( e ) IL-22 were measured by luminex assays. Error bars represent means ± s.e.m. n = 3-5 for a-d. n = 17 for human ATB patients for e. ND, not detected; UI, uninfected; UN, untreated. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 by ( a,b ) one-way ANOVA, ( c,d ) Student's t test ( e ) two-way ANOVA

Article Snippet: Human peripheral blood mononuclear cells (PBMCs) from ATB patients were isolated by Ficoll Hypaque gradient (Lymphoprep; Axis-Shield POC AS, Oslo, Norway ).

Techniques: Infection, Isolation, Mouse Assay, Enzyme-linked Immunosorbent Assay, Luminex