Journal: Cytokine

Article Title: Cloning and expression of porcine Colony Stimulating Factor-1 (CSF-1) and Colony Stimulating Factor-1 Receptor (CSF-1R) and analysis of the species specificity of stimulation by CSF-1 and Interleukin 34

doi: 10.1016/j.cyto.2012.08.008

Figure Lengend Snippet: Activity of porcine and human recombinant CSF-1 on canine and feline bone marrow progenitor cells. Canine and feline bone marrow cells were cultured with either 10 4 U/ml rh-CSF-1, 300ng/ml porcine E.Coli expressed CSF-1 or no growth factors. By day 5 of differentiation, canine and feline BMDMs with no growth factors were dead (canine A & feline D). By day 5 of differentiation for canine and day 12 for feline, BMDMS were attaching to the culture dish when cultured with either rh-CSF-1 (canine B & feline E) or porcine CSF-1 (canine C & feline F).

Article Snippet: Recombinant mouse and human IL-34 are both commercially available (R&D Systems).

Techniques: Activity Assay, Recombinant, Cell Culture

Journal: Cytokine

Article Title: Cloning and expression of porcine Colony Stimulating Factor-1 (CSF-1) and Colony Stimulating Factor-1 Receptor (CSF-1R) and analysis of the species specificity of stimulation by CSF-1 and Interleukin 34

doi: 10.1016/j.cyto.2012.08.008

Figure Lengend Snippet: Activity of recombinant human and mouse IL-34 on porcine CSF-1R expressed in Ba/F3 cells. An MTT cell viability was used to assess the biological activity of human and mouse IL-34 on expressed porcine CSF-1R. (A) Both human and mouse IL-34 are biologically active on the porcine receptor. (B) Human IL-34 has demonstrates similar activity to rh-CSF-1 on the porcine CSF-1R in an MTT assay.

Article Snippet: Recombinant mouse and human IL-34 are both commercially available (R&D Systems).

Techniques: Activity Assay, Recombinant, MTT Assay

Journal: The Journal of Experimental Medicine

Article Title: Activating receptors promote NK cell expansion for maintenance, IL-10 production, and CD8 T cell regulation during viral infection

doi: 10.1084/jem.20082387

Figure Lengend Snippet: IL-10 production by NK cells from MCMV-infected Prf1 −/− mice. (A) The top dot plots show the purity of NK cells (CD49b + ) and non–NK cells (CD49b − ) isolated from day 4 MCMV-infected wt and Prf1 −/− mice after CD49b + magnetic isolation. The numbers indicate percentages of cells in each box. Cytokine production of IFN-γ and IL-10 in the serum of infected wt and Prf1 −/− mice, or in 24 h conditioned media prepared with total splenic leukocytes and isolated subsets were determined. (B) The top dot plots show purity of NK cells (CD49b + Ly49H + ) and non–NK cells (CD49b − Ly49H − ) cells of day 4 MCMV-infected Prf1 −/− mice after flow sorting. The numbers indicate percentages of cells in each box. Cytokine production of IFN-γ and IL-10 in serum of infected Prf1 −/− mice, or in 24 h conditioned media from total splenic leukocytes and isolated subsets were determined. (C) Expression of IL-10 , IL-21 , and IL-15 in the total and sorted populations from day 4 MCMV-infected Prf1 −/− mice was analyzed by semiquantitative RT-PCR. Gapdh was used as an internal control. (D) Total splenic leukocytes, highly purified NK, and non–NK subsets from day 4 MCMV-infected Prf1 −/− mice were stimulated either on anti-Ly49H antibody or with cytokines. After 24 h of incubation, IL-10 production was determined in conditioned media. Results are representative of at least two independent experiments using a total of two to four individual mice. The studies in A were repeated twice for wt and four times for Prf1 −/− mice. Those in B were repeated three times, and those in C were repeated twice. The studies in D with anti-Ly49H antibody were repeated twice, and those with cytokines were repeated four times. The dashed white line indicates that intervening lanes have been spliced out. φ, not detected.

Article Snippet: For further stimulation, 10 5 highly purified cells were incubated either on control antibody (mouse IgG1)– and anti-Ly49H antibody–coated plates or with 100 ng/ml of mouse recombinant IL-21 (>97% purity; R&D Systems) in the presence or absence of 50 ng/ml IL-15 (>95% purity; R&D Systems).

Techniques: Infection, Isolation, Expressing, Reverse Transcription Polymerase Chain Reaction, Purification, Incubation