Immunology / Inflammation

The immunology / inflammation services are offered in co-operation with VivaCell.

A wide portfolio of protocol to evaluate the natural and specific immune response in isolated human and murine lymphoid cells and also in transformed cell lines is offered. In addition, animal models mimicking human pathologies are also available. These in vitro and in vivo models are suitable for preclincal validation of anti-inflammatory and immunomodulatory compounds.

Immunology / Inflammation: in vitro Services

Models

• primary human monocytes (rheumatic diseases, wound healing, etc.)
• primary T- / B-lymphocytes, neutrophils, Natural Killer cells
• primary fibroblasts, keratocytes, melanocytes
• primary endothelial cells (heart) and HUVEC
• murine thymocytes and primed T-cells from spleen and lymphatic nodes of antigen-stimulated mice
• enrichement of dendritic cells
• chondrocytes
• transformed T-cell lines, B-cell lines and macrophage cell lines
  
  

Parameters

• Prostaglandins (PGE2), cyclooxygenases (COX-1, COX-2), 5-lipoxygenase and determination of leukotriens by ELISA, activity of cPLA2
• cytokines by ELISA or RT-PCR (monocytes: TNF-a, IL-6, IL-1, IL-8, etc.; T-cells: IL-2, IL-4, j-IFN)
• iNOS / NO
• growth factors
• proliferation / activation of T- / B- lymphocytes
• phagocytic activity
• coagulation assays (aPTT, PT, TZ, Fxa)
• matrix metalloproteases (MMPs: MMP1, MMP3, MMP9, etc.)
• transcription factors (NF-kB, NF-AT, AP-1, OCT-1 and STATs)
• signaling studies, MAPKs (ERK 1/2, p38, JKN / SAPK); PKCs; calcineurin dephosphorylation; cyclins activation, tyrosine kinases and the like
• determination of phenotypes (CD determination by FACS analysis): single and double staining
• proliferation and cell cycle analyses by incorporation of 3[H]TdR/BrdU and / or FACS analysis:
  (a) mitogens (PHA, ConA, LPS)
  (b) TCR specific, CD3 or CD3 / CD28 or with the superantigen SEB
  (c) cell cycle analyses: progression to the S-phase and G2/M of the cell cycle
• antigen specific proliferation in murine primed T-cells, Th1 and Th2 profile by cytokine release
• allogenic T-cell prolieration (mixed lymphocyte cultures)
• induction of antigen-specific antibody production in vitro
• Natural Killer aactivity and CTL activity
• determination of cell death in primary cells by determining either necrosis or apoptosis (type I and II)
• signalling studies in primary T- / B-cells and in lymphoid tumor cells: calcium mobilisation, intracellular pH changes, and the like
• transfections in primary T-cells and in lymphoid cell lines: functional studies of transcriptional regulation using a big panel of cellular and synthetic promoters cloned in front of the luciferase gene (NF-kB-Luc, AP-1-Luc, NF-AT-Luc, IL-2, TNF, IL-6, ICAM-1, CD69, cyclins, p21, Bax, PECAM, etc.)
• stably transfected cell lines with the luciferase gene driven by several inducible promoters regulated by PMA, IL-1 and TNFa-transcripton factors (NF-kB, LTR-HIV)
  

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