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IMMUNOBIOLOGY

IMMUNOBIOLOGY. And EXPERIMENT. Immunopharmacology : intersection of immunology and pharmacology . . Focuses on drugs that affect the immune system , whether to suppress it, activate it, or manipulate it in some way. .

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IMMUNOBIOLOGY

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  1. IMMUNOBIOLOGY And EXPERIMENT

  2. Immunopharmacology: intersection of immunology and pharmacology. Focuses on drugs that affect the immune system, whether to suppress it, activate it, or manipulate it in some way. The most well-known immunopharmacology agents include anti-rejection drugs and vaccines.

  3. Fluorescence-activated cell sorting (FACS) Flowcytometry using a BD FACS Calibur.

  4. Flow-FISH (fluorescent in-situ hybridization) is a cytogenetic technique to quantify the copy number of specific repetitive elements in genomic DNA of whole cell populations via the combination of flow cytometry with cytogenetic fluorescent in situ hybridization staining protocols

  5. P53 ; RT-PCR

  6. Immunocytochemistry vs. immunohistochemistry Immunocytochemistry differs from immunohistochemistry in that the former is performed on samples of intact cells that have had most, if not all, of their surrounding extracellular matrix removed. This includes cells grown within a culture, deposited from suspension, or taken from a smear. In contrast, immunohistochemical samples are sections of biological tissue, where each cell is surrounded by tissue architecture and other cells normally found in the intact tissue. Counterstains After immunohistochemical staining of the target antigen, a second stain is often applied to provide contrast that helps the primary stain stand out. Many of these stains show specificity for discrete cellular compartments or antigens, while others will stain the whole cell. Both chromogenic and fluorescent dyes are available for IHC to provide a vast array of reagents to fit every experimental design, and include: hematoxylin, Hoechst stain and DAPI are commonly used.

  7. AktivitasSenyawaSemisintetikKuasinoiddariBuahMakasar(Bruceajavanica[L.] Merr) sebagaiAntikankerdengan Target Protein P53, Bcl-2, Kaspase-3, COX-2 dan c-Myc COX-2 AbPrimer P21 ;C-myc; Bcl2 Microscope

  8. Components of the immune system • White blood cells • Lymphoid organ • Primary lymphoid organ: • Bone marrow & fetal liver • origin of all immune cells • site for development and education of B cells • Thymus: • site for development and education of T cells • Secondary lymphoid organ • Lymph nodes, spleen, lymphoid tissue • induction sites for immune responses • Body tissues • effector sites for immune responses

  9. cells All white blood cells originate from the bone marrow Growth and differentiation factors (cytokines) produced by and present on bone marrow stromal cells determine the type of white blood cell that will emerge, as well as their relative numbers.

  10. cells Blood cells derived from bone marrow cells Adaptive imm Innate imm

  11. cells Blood cells migrate through blood and lymph nodes or home to tissues

  12. Molecules involved for antigen recognition • B cell receptor & product • antibodies (Abs): immunoglobulin (Ig) • T cell receptor (TCR) • TCR a/b (type II),g/d (type I) • Major histocompatibility complex (MHC)/HLA • Class I • Class II

  13. Immune responses • Natural/innate/non-specific • Humoral: type I Interferon (IFN-a/b), lysozyme, complement (C) • Cellular: phagocytes, NK cells • Adaptive/acquired/specific • Humoral: Abs: IgM, IgG, IgA, IgE, IgD • Cellular: T cells: • CD4+ Th,CD8+CTL, CD4+CD25+ T reg.

  14. Lymphocytes: the B and T cells

  15. The cells in adaptive immune responses • Antigen specific lymphocytes • Effector cells • Specialized accessory cells

  16. Lymphocytes • Capable of specifically recognizing and distinguishing different antigenic determinants • Responsible for the defining characteristics of adaptive IR i.e. - specificity - memory

  17. Normal Blood Cell Counts

  18. PCR , IHC, ELISA, FACS-Flowsitometri Experiment Design: Invitro; Invivo

  19. MAJOR STEPS IN IMMUNE RESPONSES CD8 T cell Antigen IL-2 cytotoxic T cells 1 4 3 2 primed CD4 T helper cell IL-2 IL-1 antigen presenting cell (macrophage, dendritic cell) CD4 T helper cell plasma cells 4 IL-2 B cell

  20. SITES OF ACTION OF IMMUNOSUPPRESSIVE DRUGS CD8 T cell 4 Antigen IL-2 X cytotoxic T cells 1 E X C A X 3 2 primed CD4 T helper cell X X D D IL-2 IL-1 B antigen presenting cell CD4 T helper cell 4 plasma cells X cytokines

  21. Immunostimulatory Cytokines • Interleukins • IL-2 (enhance antitumor actions of cytotoxic T cells and NK cells) • Colony Stimulating Factors • G-CSF (neutropenia) and GM-CSF (bone marrow transplant patients) • Interferons (uses) • alpha (anticancer uses) • beta (relapsing type multiple sclerosis) • gamma (chronic granulomatous disease)

  22. Interferon Uses • Interferon Alpha (prod. by leukocytes) • (antiviral, antiproliferative) • malignant melanoma, renal cell carcinoma, hairy cell leukemia, Kaposi’s sarcoma • Interferon Beta (prod. by fibroblasts) • (antiviral, antiproliferative) • relapsing type MS • Interferon Gamma (prod. by lymphocytes) • (stimulates NK cells and macrophages) • chronic granulomatous disease

  23. Other Hematopoetic Growth Factors • Erythropoietin alpha (Epoetin alpha) (Procrit®) • Produced by recombinant DNA technology • Stimulates division and differention of erythroid progenitor cells • Used for anemia due to renal failure or cancer chemotherapy • Adverse effects include hypertension, headache, hypersensitivity reactions are rare • Darbopoetin alpha (Aranesp®) • Recombinant long-acting erythropoetin (3X epoetin)

  24. Cytokine Inhibitors • TNF inhibitors (disease modifiers to treat rheumatoid arthritis) • Etanercept (Enbrel) • Recombinant version of TNF receptor • Infliximab (Remicade) • Chimeric human/murine anti-TNF monoclonal antibody • Anakinra (Kineret) • Human IL-1 receptor antagonist • Disease modifier agent for Rheumatoid arthritis

  25. Other Immunostimulants • Thymic Hormones • Improve primary immune deficiency in children • Synthetic Stimulants • Levamisole stimulates phagocytosis and T cell production of cytokines • Adjuvants of bacterial origin • BCG is viable strain of Mycobacterium bovis that enhances macrophage activity • BCG used for bladder cancer and melanomas

  26. Targeted Immunotherapy • Antibody-mediated delivery systems • Radiolabeled antibodies • Types of antibodies in trials • Anti-CD20 for B cell lymphomas • Anti-vascular endothelial cell growth factor • Anti-fibroblast growth factor • Anti-body to F19 on surface of activated fibroblasts

  27. New Approaches for Tolerance • Interference with costimulatory signals required for T cell activation • Two signals required for T cell activation • Signal 1 via T cell receptor • Signal 2 via costimulatory receptor-ligand pair • Antibodies to costimulator receptors (on T cell) or ligands (on antigen presenting cell) • Anti-CTLA4 (blocks B7 binding to T cell CD28) • Anti-CD40 (inhibits macrophage and endothelial activation by blocking T cell CD40 ligand binding to macrophage CD40)

  28. TUMOR CELL

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