Fagocitoza

Fagocitoza ( - a degera i - celul) este

proprietatea unor celule din organism de a ingloba in
protoplasma lor microorganisme sau particule inerte si
de a le distruge prin digerare.
Fagocitoza reprezinta unul din cele mai importante mijloace
de aparare ale organismului fata de infectii.
Prin fagocitoza, germenii patrunsi in organism sunt inglobati
si distrusi imediat chiar la poarta de intrare, nemaiputand
astfel declansa boala.
Fagocitoza reprezinta unul din fenomenele principale care se
petrec in cazul inflamatiei.
Fagocitoza a fost descoperita si descrisa in 1892 de catre
savantul rus Mecinikov.

A. Celule fagocitare
1. Neutrofile/Celule polimorfonucleare

PMN sunt celule fagocitare cu o motilitate nalt i cu nuclei

lobat. Pot fi identificate datrorit structurei nucleului sau
antigenului CD66 prezent pe suprafaa MC. Conin 2 tipuri
de granulaii care determmin proprietile antimicrobiale.
1. Granulaiile primare (azurofile) conin mieloperoxidaz,
hidrolaze acide degradative (proteaze, fosfataze,
nucleotidaze) i substane cu aciune bactericid (lizozim,
lactoferin, elastaza, catepsina G).
2. Granulaiile secundare sunt mai mici, mai puin electronodense i conin fosfataz alcalin, lizozim, NADPH oxidaza.
Nu conin peroxidaz.

Natural anti-microbial expression in the

human neutrophil. Neutrophils contain two
types of granules that contain natural antimicrobials. The azurophil granules contain
-defensins while specific granules contain
lactoferrin and the cathelicidin, hCAP-18.
Lysozyme is present in both granule types.
King et al. Reproductive Biology and
Endocrinology 2003 1:116 doi:10.1186/14
77-7827-1-116

FAGOCITE:
Neutrofilele
fagocitoz,
moarte intracelular,
inflamaie,
distrucii tisulare
Nucleu

i citoplasm
caracteristice
Granulaii
Marker

membranar CD67

Markeri pentru neutrofile

CD13

-aminopeptidaza N
metabolism al peptidelor reglatoare
CD33 -acid sialic, molecul de adeziune

MPO mieloperoxidaza
Protein bactericid

Granulaiile neutrofilelor
primare

secundare

azurofile; caracteristice
pentru neutrofilele tinere

specifice neutrofilelor mature

Conin proteine cationice,

lizozim, defensine,
proteaze i
mieloperoxidaz

Conin lizozim, NADPH

oxidaza, lactoferina i
proteina de legare a B12

A. Celule fagocitare
2. Macrofagii (monocite) celule fagocitare, cu nucleul n form
renal.
Pot fi identificai dup structura morfologic sau dup prezena
markerului CD14 pe MC. Nu conin granulaii, dar au un
coninut sporit de lizozim.

Macrofagele
-fagocitoz,
-moarte intra i extracelular,
-reparaie tisular,
-prezentarea antigenului n
cadrul rspunsului imun
specific
Nucleu

caracteristic
Marker membranar: CD14
(receptor LPS)

B. Rspunsul fagocitar la infecii

PMN i monocitele din circuitul sanguin rspund la semnalele SOS

generate de focarul de infecie.
Semnalul SOS include:
1. N-formil-metionina eliberat de bacterii
2. Peptidele sitemului de cuagulare
3. Produsele complementului
4. Citokinele eliberate de macrofagele tisulare care au ntlnit
bacteriile n esuturi.
Odat ptrunse n spaiile intercelulare unele din semnalele SOS
atrag fagocitele spre focarul de infecie prin mecanismul de
chemotaxie.

http://www.cellsalive.com/chemotx.htm

Semnale trigger
N-formil metionina
Factorii coagulrii
Produii
complementului

Rspuns fagocite
Aderare vascular
Diapedez
Chemotaxie
Activare
Fagocitoz i moarte

C. Iniierea fagocitozei
Pe suprafaa MC fagocitele au numeroi receptori prin intermediul crora
agenii infecioi se leag de celul.
1. Fc receptori bacteriile cu IgG fixai pe suprafaa lor au regiunea Fc a
anticorpilor liber, aceasta poate forma legturi cu receptorul de pe
suprafaa fagocitelor.
2. Receptorii complementului celulele fagocitelor au receptori pentru al
treilea component al complimentului, C3b

3. Receptorii Scavenger grup de receptori foarte divers care recunosc

moleculele de lipoproteine, capteaz moleculele cu sarcin negativ
4. Receptori Toll-like fagocitele posed un spectru larg de Toll-like
receptori (Pattern Recognition Receptors or PRRs) care recunosc un vast
areal de paterne moleculare (pathogen associated molecular patterns
PAMPs) ale agenilor infecioi. Legarea agenilor patogeni de ctre
receptorii Toll-like provoac eliberarea citokininelor de ctre fagocit (IL-1,
TNF-alfa i IL-6).

D. Fagocitoza etapa imediat urmtoare dup fixarea bacteriei extinderea

pseudopodelor de cte fagocit n jurul bacterie.
Bacteria este inglobat cu formarea fagozomului.

Granulaiile sau lizozomii

fagocitului fuzioneaz cu fagozomul.
Ca rezultat, bacteria este cuprins n fagolizozom.
http://www.cellsalive.com/mac.htm

http://www.microbelibrary.org/images/tterry/anim/phago053.html

E. Introducerea stresului
oxidativ i degradarea
intracelular
Stresul oxidativ eliberarea rapid a speciilor reactive de oxigen (radical
superoxid, peroxidul de hidrogen) de ctre diferite celule.
n timpul fagocitozei se atest o majorare a consumului de glucoz i
O2 care se reflect n stresul oxidativ.
n consecin - producerea compuilor ce conin oxigen
Scopul distrugerea intracelular oxigen-dependent a bacteriilor
fagocitate

Bacteriile pot fi distruse i de substanele deja existente eliberate de

ctre granule sau lizozomi degradare intracelular oxigenindependent.
http://www.cellsalive.com/nbt.htm
http://www.youtube.com/watch?v=irtc01i2WPA

I. Degradarea intracelular oxigen-dependet mieloperoxidaz-independent

Pe parcursul fagocitozei glucoza este metabolizat prin untul pentozo-fosforic cu
formarea NADPH.
Citocromul B, parte component a granulelor, se combin cu NADPH i l activeaz.
NADPH oxidaz activ utilizeaz oxigenul pentru oxidarea NAPDH.
Ca rezultat producerea anionului de superoxid.
A) O parte de anioni de superoxid sunt convertii H2O2 i oxigen atomar de ctre
SOD (superoxiddismutaza).
B) Supeoxid anionul poate interaciona H2O2 cu formarea radicalului hidroxil i
oxigen atomar.
Rezultatul reaciilor producerea speciilor reactive de oxigen - anionul superoxid
(O2-), peroxidul de oxigen, oxigenul atomar (1O2) i radicalul hidroxil (OH).

II. Degradarea intracelular oxigen-dependet mieloperoxidazdependent

Odat ce granulaiile azurofile fuzioneaz cu fagozomul, mieloperoxidaza
este eliberat n fagolizozom.
Mieloperoxidaza utilizeaz H2O2 i ionii de halogen (cel mai frecvent Cl-)
pentru producerea de hipoclorid substan cu un grad sporit de
toxicitate.
Ionii de hipoclorit pot spontan degrada cu formare de oxigen atomar. Ca
urmare se produc anionul de hipoclorid toxic (OCl-) i oxigen atomar
singlet oxygen (1O2).

Cile de degradare intracelular

Degradare intracelulara
oxigen-dependenta
mieloperoxidaza-independenta

oxigen-independenta

mieloperoxidaza-dependenta

Mediatori ai degradrii oxigen

independente n fagolizozom
Molecule efectoare

Funcie

Proteine cationice (catepsina)

Alterarea membranei
microbiene

Lizozim

Hidroliza mucopeptidelor
din peretele bacterian

Lactoferina

Extrage fierul bacterian

Enzime hidrolitice (proteaze)

Diger organismele distruse

III. Reaciile de detoxifiere

PMN i macrofagii au mecanisme de autoprotecie de ctre
speciile reactive de oxigen.
Reacia presupune dismutarea anionului de supeoxid n
peroxidul de hidrogen de ctre SOD i conversia peroxidului n
ap de ctre catalaz.

Reacia

Enzima

H2O2 + Cl- --> OCl- + H2O

OCl- +

H2O -->

1O

+Cl- +

H2O

Mieloperoxidaza

2O2 + 2H+ --> O2- + H2O2

Superoxiddismutaza

H2O2 --> H2O + O2

Catalaza

The main molecules and transporters thought to participate in charge compensation and pH
regulation during the respiratory burstA phagocyte is depicted engulfing a bacterium into a
nascent phagosome, which will proceed to close and become intracellular.

DeCoursey T E Physiology 2010;25:27-40

2010 by American Physiological Society

IV. Degradarea intracelular oxigen-independent

catepsina eliberat n fagolizozom poate distruge MC a bacteriei

lizozimul distruge PC
lactoferina helateaz ionii de Fe
enzime hidrolitice distrug proteinele bacteriene

Thus, even patients who have defects in the oxygen-dependent killing

pathways are able to kill bacteria. However, since the oxygen-dependent
mechanisms are much more efficient in killing, patients with defects in
these pathways are more susceptible and get more serious infections.

Oxygen-independent mechanisms of intracellular killing

Effector Molecule
Cationic proteins (including cathepsin)
Lysozyme
Lactoferrin
Proteolytic and hydrolytic enzymes

Function
Damage to microbial membranes
Splits mucopeptide in bacterial cell wall
Deprives proliferating bacteria of iron
Digestion of killed organisms

Distrugerea bacterian NO-dependent

Interaciunea bacteriei cu receptorii de tip Toll duce la sinteza
TNF-alfa (Tumor necrosis factor - citokinin ce particip n
procesul inflamator). Acesta din urm induce expresia genei NOsintetezei i drept rezultat se produce NO.
Oxidul nitric eliberat de ctre celul este toxic i poate distruge
MO din vecintatea macrofagului.

V. CELULE UCIGAE NESPECIFICE

A. NK i LAK
Natural killer (NK) cells are also known as large granular lymphocytes (LGL)
because they resemble lymphocytes in their morphology, except that they are
slightly larger and have numerous granules. NK cells can be identified by the
presence of CD56 and CD16 and a lack of CD3 cell surface markers. NK cells are
capable of killing virus-infected and malignant target cells but they are relatively
inefficient in doing so. However, upon exposure to IL-2 and IFN-gamma, NK cells
become lymphokine-activated killer (LAK) cells, which are capable of killing
malignant cells. Continued exposure to IL-2 and IFN-gamma enables the LAK
cells to kill transformed as well as malignant cells. LAK cell therapy is one
approach for the treatment of malignancies.
How do NK and LAK cells distinguish a normal cell from a virus-infected or
malignant cell? NK and LAK cells have two kinds of receptors on their surface a
killer activating receptor (KAR) and a killer inhibiting receptor (KIR). When the
KAR encounters its ligand, a killer activating ligand (KAL) on the target cell the
NK or LAK cells are capable of killing the target. However, if the KIR also binds to
its ligand then killing is inhibited even if KAR binds to KAL. The ligands for KIR
are MHC-class I molecules. Thus, if a target cell expresses class I MHC
molecules it will not be killed by NK or LAK cells even if the target also has a KAL
which could bind to KAR. Normal cells constitutively express MHC class I
molecules on their surface, however, virus infected and malignant cells down
regulate expression of class I MHC. Thus, NK and LAK cells selectively kill virusinfected and malignant cells while sparing normal cells.

B. K cells (Figure 14)

Killer (K) cells are not a morphologically distinct type of cell. Rather a K
cell is any cell that mediates antibody-dependent cellular cytotoxicity
(ADCC). In ADCC antibody acts as a link to bring the K cell and the
target cell together to allow killing to occur. K cells have on their surface
an Fc receptor for antibody and thus they can recognize, bind and kill
target cells coated with antibody. Killer cells which have Fc receptors
include NK, LAK, and macrophages which have an Fc receptor for IgG
antibodies and eosinophils which have an Fc receptor for IgE
antibodies.

Table 5. Characteristics of cells involved in non-specific

resistance
Identifying marker(s) and/or function
Ig

Fc

CD

Phago
cytosi
s

IgG
IgG
IgG
IgG
?
IgE

CD67
CD14
CD56 & 16
?
?
CD67

+
+
?
-

CD3
Effector cell
Neutrophil
Macrophage
NK cell
K-cells
LAK cell
Eosinophil

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http://2008.igem.org/Team:Caltech/Project/Oxidative_Burst

http://krebbing.blogspot.com/2006/12/respiratory-burst.html