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Doc. dr. Tomaž Bratkovič, mag. farm.
Univerza v Ljubljani, Fakulteta za farmacijo,
Aškerčeva 7, 1000 Ljubljana
POVZETEK
ABSTRACT
3
3.2.2
od limfocitov T odvisna aktivacija limfocitov B
Slika 1: Dve poti proženja tvorbe protiteles proti proteinski učinkovini (prirejeno po (12)). A. Od limfocitov T odvisna aktivacija limfocitov B:
Antigen predstavitvena celica (APC) fagocitira protein [1], ga razgradi na krajše antigenske peptide, ki jih predstavi na svoji površini vezane na
poglavitni histokompatibilnostni kompleks razreda II (major histocompatibility complex class II, MHC II) [2]. Naivna T-celica pomagalka (Th) s
T-celičnim receptorjem (TCR) »prepozna« pripadajoči T-celični epitop in sočasno prejme še dodatne aktivacijske signale (posredovane s
CD80/86 na CD28), kar sproži aktivacijo in proliferacijo Th [3]. Naivni limfocit B »prepozna« B-celični epitop enakega antigena z B-celičnim
receptorjem (BCR), antigen internalizira [4], razgradi in predstavi T-celične epitope na MHC II [5]. Slednje »spoznajo« predhodno aktivirani
limfociti Th, ki spodbudijo proliferacijo specifičnih limfocitov B in njihovo diferenciacijo do plazmatk in dolgoživih spominskih limfocitov B.
Podobno kot pri aktivaciji Th tudi aktivacija limfocitov B zahteva dodatno stimulatorno signalizacijo (vezavo CD40L na CD40). B. Neposredna
(od limfocitov T neodvisna) aktivacija limfocitov B: Ponavljajoči epitopi na površini proteinskega agregata so odgovorni za združevanje BCR
na površini naivnih limfocitov B, kar sproži znotrajcelično signalizacijo, ki vodi v aktivacijo in diferenciacijo do plazmatk. Kostimulatorni signali v
obliki ligandov receptorjev TLR tudi prispevajo k aktivaciji, proliferaciji in diferenciaciji limfocitov B, podobno velja za nekatere citokine, ki jih
izločajo naravne celice ubijalke in limfociti T. LPS – lipopolisaharid.
Figure 1: Two pathways leading to anti-drug antibodies (adopted from (12)). A. T-cell dependent B-cell activation: Antigen-presenting cell
(APC) takes up a protein by phagocytosis, processes it and displays short antigenic peptides bound to major histocompatibility complex II
(MHC II). Naïve helper cell (Th) “recognizes” the cognate T-cell epitope via T-cell receptor (TCR). Concurrent stimulatory signals (conveyed by
CD80/86 binding to CD28) trigger activation and proliferation of Th lymphocytes. Naïve B-cell “recognizes” the B-cell epitope of the same
antigen via B-cell receptor (BCR), internalizes the antigen, processes it and displays T-cell epitopes bound to MHC II. These, in turn, are
“recognized” by previously activated Th-cells, responsible for proliferation of specific B-cells and inducing their differentiation into antibody-
producing plasma cells and long-lived memory B-cells. As with Th-cell activation, activation of B-cells requires additional stimulatory
signaling (binding of CD40L to CD40). B. Direct (T-cell independent) activation of B-cells: Epitope arrays on the surface of protein aggregates
induce clustering of BCRs on naïve B-cells, leading to intracellular signaling, resulting in activation and differentiation into plasma cells. Co-
stimulatory signals, such as TLR ligands, contribute to the activation, proliferation and differentiation of B-cells, as do cytokines secreted by
natural killer cells and T-cells. LPS – lipopolysaccharide.
Neposredna aktivacija limfocitov B
pat-
tern-recognition receptorsToll-like
receptors
porušenja tolerance
danger si-
gnals
3.2.2
Chinese hamster ovary cells
Escherichia coli
E. coli
E. coli
Slika 2: Interakcije proteinskih agregatov z antigen predstavitvenimi celicami (APC) (prirejeno po (12)): Receptorji TLR vežejo agregate, ki
spominjajo na molekularne vzorce, povezane s patogeni (pathogen-associated molecular patterns, PAMP). Receptorji komplementa (CR)
prepoznavajo s komplementom opsonizirane (gliko)proteinske agregate. Na lektinske receptorje (LR) se vežejo glikoproteini s telesu tujimi
sladkorji. Agregati terapevtskih protiteles se lahko vežejo na receptorje za imunoglobulinsko regijo Fc (FcγR). Opisane interakcije so pogoj za
receptorsko posredovano fago- in endocitozo (ni prikazano) kot tudi proženje signalizacijskih poti, ki vodijo v aktivacijo antigen predstavitvenih
celic in s tem sproščanje citokinov ter povišano izražanje MHC II in ligandov kostimulatornih receptorjev (npr. CD80 in CD86). Ti procesi
prispevajo k aktivaciji naivnih limfocitov Th in posledični aktivaciji limfocitov B (glejte sliko 1).
Figure 2: Interactions of (glyco)protein aggregates with antigen-presenting cells (APC) (adopted from (12)): TLR receptors bind aggregates
mimicking pathogen-associated molecular patterns (PAMP). Complement receptors (CR) interact with complement-opsonized aggregates.
Lectin receptors (LR) bind foreign saccharides. Aggregates of therapeutic antibodies are bound by Fc receptors (FcγR). In addition to
providing stimulatory signals via receptor binding, aggregates are taken up by receptor-mediated phago-/endocytosis (not shown). APC
activation results in cytokine secretion and upregulation of MHC II and co-stimulatory receptors (e.g., CD80 and CD86). The described
processes contribute to activation of naïve Th-cells and (indirectly) B-cells (see Figure 1).
3.2.2
3.2.3
4
in vitro
HLA
3.2.4
in silico
in vitro
in vitro
HLA
Slika 3: Incidenca pojava protiteles, usmerjenih proti infliksimabu,
med 72-tedensko raziskavo pri pacientih s Crohnovo boleznijo, ki so
prejemali zdravilo občasno ali v rednih intervalih (tj. kontinuirano),
bodisi kot monoterapijo ali v kombinaciji z imunosupresivom ((33),
povzeto po (31)).
Figure 3. Incidence of anti-infliximab antibodies at any time during
72-week-trial in Crohn disease patients receiving the drug as
monotherapy or in combination with an immunosuppressive, either
episodically or continuously ((33), adopted from (31)).
baseline signal
spremem-
bah proizvodnega procesa
primerljiva
Podobna biološka zdravila
similarity exercise
podobno
avtomatična
JAZMP pod-
pira uvajanje PBZ [podobnih bioloških zdravil], ker mora
biti podobnost oz. primerljivost PBZ v razmerju do zadevnih
referenčnih bioloških zdravil z vidika kakovosti, varnosti in
učinkovitosti dokazana v okviru postopka pridobitve dovo-
ljenja za promet za vsako PBZ. […] Odločitev o njihovi
uporabi oz. njihovo zamenjevanje (terapevtski preklop) lahko
poteka v le v kliničnem oz. ambulantnem okolju in mora
biti predmet medicinske obravnave vsakega posameznega
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