Passive Immunisierung und Antikörpertherapien

1. Passive Immunisierung und Antikörpertherapien Konteptvorlesung 3 Themenblock8 Beda M. Stadler Institut für Immunologie Diese Folien stehen auch als PPT Files zum download bereit unter: http://www.iib.unibe.ch/wiki/?p=teaching/medical_students&l=de KV 8.3

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4. Top Ten Monoclonal Antibodies2009/2010 Thearapeutische Antikörper auch zu teuer für uns?

5. Übersicht / Lernziele • Antikörper Produktion einst • Passive Immuntherapie einst • Monoklonale / rekombinante Antikörper • Antikörper als Therapie für: • Immunsuppression • Krebs Therapie • Immunmodulation • Anti-Adhäsionstherapien • Rückblick und Zukunft Paul Ehrlich vor 100 Jahren KV 8.3

6. Passive Immuntherapie einst KV 8.3

7. Krankheit Therapeutikum Schwarze Spinnenbiss Pferd Antivenin Botulismus Pferd Antitoxin Diphtherie Pferd Antitoxin Hepatitis A and B Multispender human Gammaglobulin Masern Multispender human Gammaglobulin Tollwut Multispender human Gammaglobulin Schlangenbiss Pferd Antivenin Tetanus Multispender human Gammaglobulin oder Pferd Antitoxin Passive Immuntherapie mit polyklonalen Antikörpern KV 8.3

8. Monoklonale AntikörperHybridoma Technologie KV 8.3

9. Therapeutische Maus Antikörper Probleme • Immunogenizität(humane anti-Maus AK, HAMA) • keine Effektor Mechanismen(Maus Fc) • Unerwartete Toxizität(Reaktivitiät mit normalem Gewebe, Kreuzreaktion mit anderen Antigenen) • Verlust von Tumorantigen(z.B. anti-idiotyp Behandlung von Lymphomen) KV 8.3

10. Immunogenicität von Infliximab(chimärermonokl. AkgegenTNFα) • 61% of patients had detectable antibodies after the fifth infusion of Infliximab • Incidence of the formation of antibodies to Infliximab is reduced with immuno-suppressive therapy Human anti-mouse Ab Human/mouse chimeric Ab (Infliximab) KV 8.3

11. Diversity of Antibodies is Generated by Gene Rearrangement Dank Wissen um Grundlagen rekombinante Antikörper KV 8.3

12. scFv IgG Fab CL VL VH Hinge CH CH2 Fv VL VH Fc CH3 Die Domänenstruktur hilft zum Umbau von Antikörpern KV 8.3

13. Humanisierung von Maus Antikörpern Maus Chimäre Humanisiert Human KV 8.3 Ezzell, Scientific American

14. TherapeutischeAntikörper KV 8.3

15. Fab Fv scFv dsFv VH Mono- & plurivalente Ak-Fragmente scFv Diabody Triabody Tetrabody KV 8.3

16. Transgene Humanisation humane Antikörper in human Ig transgenenen Mäusen KV 8.3

17. Der Wunsch nach humanen AK Humane Antikörper mittels Repertoire Klonierung mRNA von B Zellen Antikörper (Fab) Expression auf p3 desM13 Phagen Blut PCR of H&L chain genes Klonierung und Rekombination mRNA  cDNA KV 8.3

18. Expression von Antikörpern Antikörper DNA Transfektion Antikörper Vektor Zellkultur Expression KV 8.3

19. Flask Spinner Bioreaktor (Bench scale) Bio Reactor (Plant) 1mL → 100mL → 1L → 1Lx3 → 12L → 100L → 300L → 2’000L→ 10’000L Subkultur Produktion Industrielle Antikörper Produktion KV 8.3

20. Ersatz von passivem IgGdurch rekombinante anti-RhD RhD+ Erythrozyten rekombinante Antikörper mittels phage display KV 8.3

21. Neue Therapeutische Antikörper KV 8.3

22. Immunsuppressive Anikörper(anti-T Zell Antikörper) • Lymphocyte immune globulin (Atgam®) • lytic to human thymic lymphocytes; blocks T-cell responding • Anti-CD3: (OKT3®, muromonab-CD3) • binds CD3, blocks antigen binding; depletes T-cells • Anti-CD3: huOKT3 • Humanized version of OKT3 • Anti-CD4: OKT4 • less infusion-related reaction • Anti-Tac monoclonal antibody (Zenopax®) • binds IL-2 receptor of activated T-cells causing their inactivation KV 8.3

23. Campath-1H OKT3 Lymphocyte Depleting Antibody Therapies Polyclonal Anti-Thymocyte Globulin CD52 CD3 KV 8.3

24. slg CD20 CD19 CD22 DR Immunsuppressive Anikörper(anti B-Zell Antikörper) • Anti-CD20: Rituxan® (Rituximab) • results in mature B-cell depletion and induces apoptosis (B-cell mediated vascular rejection) for B-cell Non Hodgkin’s Lymphoma • anti-CD52: Campath-1H (Alemtuzumab) • depletes T and B lymphocytes, NK cells, monocytes and macrophages KV 8.3

25. Side effects • fever • chills • weakness • headache • nausea • vomiting • diarrhea • low blood pressure • rashes KV 8.3

26. AntikörperKrebs Therapie Strategien 1 2 5 ADCC -Y-90 -Y-90 NK Anti Angiogenese Complement pathway -Y-90 Tumor cell TNF -Y-90 Anti-Idiotyp IL-1 -Y-90 Mac -Drug -Tox 3 4 -Drug -Tox -Tox -Tox -Drug KV 8.3

27. Ziel Zelle Strategie 1Antikörper vermittelte Zytotoxizität NK Zelle Ziel Zelle Apoptosis viaInduktion von intrazellulärenSignal-Pathways Ziel Zelle Antikörper-abhängige Zytotoxizität (ADCC) Complement-abhängigeZytotoxizität (CDC) KV 8.3

28. z.B. Herceptin® (Trastuzumab) normal cell tumor cell treatment Binds HER-2 (human epidermal growth factor receptor 2), a growth factor receptor found on some tumor cells (some breast cancers, lymphomas). Over-expression of HER2 causes increased cell growth and reproduction. HER2 protein over-expression affects approximately 25% to 30% of breast cancer patients KV 8.3

29. Strategie 1: maligne B-Zellen Stem cell Pro- B cell Pre- B cells Immature B cell Mature B cell Activated B cell Plasma cell Antigen independent Antigen dependent HLA-DR TDT CD19 CD10 CD20 CD22 CD21 CD38 B-cell Lymphomas (NHL, CLL) Leukemias from B-cell Precursors (B-ALL) Neoplasias: Multiple Myeloma KV 8.3

30. Strategie 1anti idiotypische Antikörper (B-Zell Lymphome) KV 8.3

31. Strategie 2: Verschiedene Isotope Nackter Antikörper Radiomarkierter Antikörper KV 8.3

32. Isotopen markierte Antikörper LymphoCide (anti CD22, found on some B-cell leukemias. Tositumomab (Anti-CD20) Lym-1 (Oncolym). Anti- HLA-DR expressed at high levels on lymphoma cells. KV 8.3

33. Strategie 3: Mylotarg® (anti CD-33) A conjugate of a monoclonal antibody that binds CD33 (cell-surface molecule expressed by the cancerous cells in acute myelogenous leukemia (AML) but not found on the normal stem cells needed to repopulate the bone marrow) and calicheamicin, an oligosaccharide that blocks the binding of transcription factors (proteins) to DNA and thus inhibits transcription. KV 8.3

34. 4 Antikörper 3 2 1 Prodrug Enzym Tumorantigen Zytotoxischer Wirkstoff Tumorzelle Strategie 4: Enzym markierte Antikörper Antibody directed enzyme prodrug therapy (ADEPT) KV 8.3

35. Strategie 5: Anti-Angiogenes KV 8.3

36. Anti-VEGF: Avastin ® • Recombinant humanized monoclonal antibody against VEGF; prevents binding with its receptor. • Blocks stimulation of angiogenesis  stops tumor growth. KV 8.3

37. Immunmodulatorische Antikörper • Anti-TNF Therapie • Infliximab • Etanercept • Humira • etc (Biosimilarika) • Anti-IgE Therapie • Xolair KV 8.3

38. Anti-TNF-a: Infliximab • Chimeric monoclonal antibody • Binds specifically to TNF-a, neutralizing its activity • Used for Crohn’s disease and rheumatoid arthritis • Side-effect: can convert a latent case of tuberculosis into active disease KV 8.3

39. TNF TNFR2 Nucleus NFB transcription No Signal Infliximab: Mechanism of Action Binds and neutralizes soluble and membrane bound TNF KV 8.3

40. C’ Complement Receptor Complement Activation Phagocyte Phagocytosis T Cell Other Postulated Mechanisms of Action… Antibody dependent cell-mediated cytotoxicity (ADCC) Lysis of TNF-expressing cells through complement (C’) activation KV 8.3

41. TNF-a: Rezeptor oder Antikörper Infliximab Etanercept KV 8.3

42. Biosimilarika: z.B. anti-TNF KV 8.3

43. OmalizumabReduces IgE and FcεRI Levels Omalizumab (Xolair®) ↓ Inflammation ↓ Symptoms ↓ free IgE ↓ FcεRI ↓ Degranulation anti-IgE FcεRI Allergic Inflammation Mediator Release B MC IgE Holgate S. et al., J Allergy ClinImmunol 2005 vol. 115 (3) pp. 459-65

44. Anti-IgE: Omalizumab (Xolair®) KV 8.3

45. Anti Adhäsionstherapie • Efalizumab (Raptiva®) • Anti-CD11a (subunit of LFA-1) inhibits activation of T cells • Alefacept (Amevive®) • binds to CD2 on memory effector T lymphocytes,inhibiting their activation • Abciximab (Reopro®) • Anti-Gp IIb/IIIa KV 8.3

46. Efalizumab (Raptiva) Interactions between leukocyte-function associatedantigen type 1 (LFA-1) and intercellular adhesion moleculesare important in the pathogenesis of psoriasis. Efalizumab,a humanized monoclonal antibody, binds to the subunit (CD11a)of LFA-1 and inhibits the activation of T cells KV 8.3

47. Anti-CD2 CD2 Alefacept (Amevive®) KV 8.3

48. Abciximab (Reopro®) Anti-GpIIb/IIIaInhibits the clumping of platelets by binding the receptors on their surface that normally are linked by fibrinogen. Helpful in preventing reclogging of the coronaryarteries in patients who have undergone angioplasty. KV 8.3

49. Ausblick: mehr Antikörper für… • Transplantation • Knochenmark, Niere, Leber und Pancreas InselZellen. • Krebs • Metastatic Brust Krebs, CLL, non-Hodgkin’s Lymphome. • Autoimmunkrankheiten • Rheumatoide Arthritis, Psoriasis, Multiple Sclerosis, Crohn’sKrankheit. • Infektionskrankheiten • Respiratory syncytial virus, Cytomegalovirus, Septikämien. • Allergien: • Mehr als 150 weitereAk in der Pipeline… KV 8.3