IMMUNOBIOLOGY

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.

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

5. 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

8. P53 ; RT-PCR

9. 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.

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

13. 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

14. 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.

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

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

17. 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

18. 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.

25. Lymphocytes: the B and T cells

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

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

28. Normal Blood Cell Counts

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

34. 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

35. 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

41. Corticosteroid Actions • Inhibition of IL-1 and TNF gene expression and synthesis • Decreased activation of T lymphocytes by decreasing IL-1 release • Decreased neutrophil functions espchemotaxis • Decreased antibody production (high doses) • Decreased release of kinins and proinflammatoryeicosanoids (prostaglandins and leukotrienes)

42. Corticosteroid Immunosuppression • Decreased cell-mediated immune reactions that mediate rejection of organ transplants • Mechanisms: • decreased activation of T lymphocytes by inhibition of IL-1 synthesis by macrophages • decreased lymphocyte mobilization out of lymphoid organs (lymphopenia)

43. TOLEROGEN (reviewed by Mowat, 1987) Oral administration of protein antigens to naive adult animals has been shown repeatedly to induce antigen-specific hyporesponsiveness of systemic immunity (oral tolerance) (reviewed by Mowat, 1987) and may play an important role in the prevention of food hypersensitivity disease. Previous studies have shown

45. The mucosal response to infection and regulation of mucosal immune responses Commensal bacteria  recognize by epithelial cells EC secretion of cytokines inhibit the expression of pro-inflammatory cytokines by DC and macrophage  DC & macrophages secrete signals for the expansion and/or survival of T cells with regulatory capacities, including regulatory T cells, T regulatory type 1 (TR1) cells, T helper 2 (TH2) cells and TH3 cells. Musketeers Course October 2008

46. 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)

47. 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

48. 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)

49. 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

50. 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