anticorpi monoclonali
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ANTICORPII
MONOCLONALI
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SISTEMUL IMUNITAR
Immunis = scutit Sistemul imunitar = totalitatea proceselor
biologice, a organelor, a celulelor, a produsilor celulari care asigura protectia impotriva infectiilor
Imunitatea Nespecifica Specifica
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ANATOMIA SISTEMULUI IMUNITAR
Amigdale si adenoide
Ganglioni limfaticiVase limfatice
Timus
Ganglioni limfatici
Splina
Placi PeyerApendice
Maduva hematogena
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CELULELE SISTEMULUI IMUNITAR
Celula stem multipotenta
Celula stem hematopoetica
Plachete
Macrofag
EritrocitePMN eozinofil
PMN neutrofil
Megacariocit
Mastocit
PMN bazofil
Limfocit T
Celula NK
Celula dendritica
Limfocit B
Celula progenitoare limfoida
Celula progenitoare
mieloida
Monocit
Maduva
Os
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ACTIVAREA LB SI PRODUCEREA DE ANTICORPI
Celula prezentatoare de Ag
Antigen
Anticorp circulant
Procesarea Ag
HLA clasa II
Antigen
Th activat
HLA clasa II si Ag procesat sunt expuse
Anticorpi
Plasmocit
Celula prezentatoare de Ag
Rec. de Ag specific LB
LBLimfokine
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ANTICORPII
Lanturi grele
Paratop
Regiune constanta
Lanturi usoare
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ANTICORPII
IgA, IgD, IgE, IgG
IgA
IgM
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PRODUCEREA ANTICORPILOR MONOCLONALI
Plasmocite producatoare de Ac
Antigen
Fuzionarea celulelor HIBRIDOM Mielom
Anticorpii monoclonali sunt purificati
Clonele dorite se cultiva sau se congeleaza
Testarea clonelor
Hibridoamele se separa si se cultiva
Cultivarea hibridoamelor
Mentinerea hibridoamelor in soareci
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PRODUCEREA ANTICORPILOR MONOCLONALI
1. Antigenul (celule, componente ale membranei celulare, microorganisme, etc.) se injecteaza soarecelui
2. Cand titrul de anticorpi serici a crescut corespunzator, se realizeaza splenectomia si se colecteaza LB
3. Se obtine hibridomul prin fuzionarea LB murine cu celule provenite din mielom
4. Se cultiva hibridoamele si se colecteaza anticorpii
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PROBLEME
HAMA = human anti-mouse antibodies Dupa 10-14 zile Eruptie cutanata (urticarie) Bronhospasm Hipotensiune Soc anafilactic
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TEHNICI DE INGINERIE GENETICA
Soareci transgenici (gene pentru Ig umane) Mab himerici =
Regiuni variabile murine Regiuni constante umane
Mab umanizati = Regiuni hipervariabile murine 95% umani
Bacteriofagi/plasmide
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APLICATIILE ANTICORPILOR MONOCLONALI
Diagnostic
Terapie
Cercetare
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APLICATIILE ANTICORPILOR MONOCLONALI
Diagnostic
Imunohistochimie Imunofluorescenta Radioimunodetectie (RIA)
Medicamente Hormoni Proteine Toxine Droguri
Imunodetectie enzimatica (ELISA)
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IMUNOHISTOCHIMIE DIRECTA
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IMUNOHISTOCHIMIE INDIRECTA
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IMUNOHISTOCHIMIE
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IMUNOHISTOCHIMIE
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IMUNOFLUORESCENTA
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IMUNOFLUORESCENTA
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APLICATIILE ANTICORPILOR MONOCLONALI
Terapie
Infliximab –TNFα Abciximab – GpIIb/IIIa Omalizumab – IgE Rituximab – CD20 Alemtuzumab – CD52 Trastuzumab – HER2 Bevacizumab – VEGF
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IMUNOTERAPIE
Anticorp
Celula tumorala mamara
Factor de crestere
Herceptin blocheaza receptorul
Cresterea se opreste
Radioizotop
Antigen
Distrugerea celulei tumorale
Herceptin
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APLICATIILE ANTICORPILOR MONOCLONALI
Cercetare
Imunohistochimie Identificarea unor molecule Purificarea unor structuri