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Apoptosis - A biological phenomenon

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Biochem. Cell. Arch. Vol. 12, No. 2, pp. 237-248, 2012 ISSN 0972-5075

Review Article
APOPTOSIS – A BIOLOGICAL PHENOMENON
Y K Lahir
Department of Zoology, Thakur College of Science and Commerce, Kandivli (E), Mumbai-400 101, India
e-mail : lahiryk@gmail.com
(Accepted 9 June 2012)

ABSTRACT – Apoptosis is a biological process under which cells undergo planned death without harming their ambient
extracellular microenvironment. This tendency on the part of cells is essential to retain ‘tissue-turn-over’ in an organ, enhance
efficiency of the Immune system, removal of those cells which have become awry or out of order. This phenomenon is accomplished
by involving effecter mechanisms; generally these mechanisms are well oriented and preserved. The components involved are
mostly maintained and prevented from being damaged. As soon as this process is over the end products, the ‘apoptotic bodies’ are
eliminated in a well organized manner without spilling the contents. The cell undergoing apoptosis can be observed by the use
of ‘DNA laddering technique’. Apoptosis plays key role in the processes such as development, differentiation, organogenesis,
homeostasis of lymphocytes and other cells. Well documented study and understanding of this process is likely to emphasize its
involvement in cytotoxicity and clinical aspects which are likely to lead to the effect and target oriented modes to tackle cancer
and related disorders.
Key words : Apoptosis, caspases, mitochondrial regulatory mode, mitochondrial apoptosis induced channel, tumor necrosis
factor, Fas receptor mode, p-53, p-73, Retinoblastoma (Rb), survival signals, death signal, cancer cell, immunity, MCH/HLA,
Thymus, dendrictic cells, cytotoxicity, interleukins, CD-95, perforin, granzyme, pathogen associated molecular pattern.

INTRODUCTION to characteristic cellular changes in the morphological,

Cells sacrifice them-selves in order to maintain a
normal growth, balance and good health of an organism.
These cells are programmed to kill them selves without
causing any injury, disturbance or discomfort to
neighboring cells. This process may be compared to the
dropping or falling of the leaves from trees during autumn
or other wise. The falling leaves have finished their
proposed function with respect to sustain the life of a
tree and quietly get detached from the tree making room
for new leaves. Even the cells of our skin on hands or
feet get detached seasonally or some of the phagocytes
are perish after they have done their function. Most
probably extra-cellular chemical messenger get attached
to the surface of cell membrane receptor, this bondage is
likely to initiate the process of apoptosis. This process
may be quoted as an apt example of signal transduction
path way in which change in magnitude of various
metabolites takes place; these changes are favorably
associated with apoptosis.
“Apoptosis is a normal phenomenon, it helps to remove
superfluous, disordered cells or cell which are triggered
to self-destruction; thus maintains normal health of the
organism as a whole.”
This process of programmed cell death (PCD) has
been observed in most of the organisms. Biochemical
events, extra cellular and intra-cellular and molecular, lead
biochemical and physiological aspects, all these result in
the death or the suicide of the cell. Among morphological
changes blebbing is prominent and is formed on the
membrane of the cell. The blebbing develops due to
localized decoupling of cytoskeleton; this process is called
“Zeiosis”. During apoptosis shrinkage of cell, nuclear
fragmentation, condensation of chromatin material and
fragmentation of chromosomal DNA take place. Albert
et al (2008) have reported the formation of fragments
during apoptosis; these fragments are called ‘apoptotic
bodies’. Later, phagocytes engulf these bodies before the
contents of cell spill on to the surrounding cells and cause
damage. These activities do not take place during
necrosis.
Role of Apoptosis
Apoptosis eliminates selected cells to ensure normal
development. During normal process of development the
cells which are not required are subjected to apoptosis.
As the embryonic development precedes some of the
undesired cells and tissues are pruned, this helps to attain
an appropriate shape and size of the tissue and organ.
This enables the normal functioning of the tissue and
organs at a given stage of life of an organism. Examples,
include pruning of the male or female reproductive
structures at primordial stage during sexual differentiation
and further development; formation of perfect digits/
238 Y K Lahir
fingers and removal of web like membrane between them
(conversion of flipper like structure in to hand or foot);
during metamorphosis in case of frog, loss of gills, tail,
shortening of intestine, changes in circulatory system etc.
Apoptosis maintains “tissue-turn over” in the organs
and systems of an adult; in an adult a specific balance is
restored between the production of number and rate of
new cells and the number and rate of self-destroyed
cells. This ratio is called tissue-turn over. The optimal
functioning of an organ and/or system depends on its
tissue-turn over. This ratio varies in different organs and/
or systems. This helps to retain the appropriate required
numbers of the cells in a given tissue and organ.
Apoptosis is effective in enhancing the efficiency of
immune system. Immune system is a natural process
involved in the removal or elimination of the infected cells.
The WBCs which become defunct and no longer effective
are eliminated.
Apoptosis removes undesired cells. The cells which
have become aged, irreparable, damaged by radiation or
toxins are removed from the tissue without harming the
neighboring and rest of the cells. Cells which become
awry i.e. they become crooked or out of order or amiss
or wrong or mutilated directly or indirectly are eliminated
from the body.
Process of Apoptosis
Apoptosis is a controlled, intentional and tidy way to
eliminate undesired and unwanted cells or any cell which
is liable to cause threat or damage to the body of an
organism. Even normal cells carrying out normal life
activity and have completed their functions but have been
rendered defunct are subjected to apoptosis for safe
elimination.
Cells undergoing apoptosis do not swell unlike necrotic
cells; generally this swelling is caused because of non-
pumping of Naz . The apoptotic cells shrink and get
isolated from the surrounding cells without causing
inflammation to them. Apoptotic cell exhibits changes,
these are metabolic and structural in nature; metabolic
changes include activation of cascades of intracellular
protein cutting enzymes (proteolytic enzymes), these
enzymes degrade the cell from within. The membrane of
mitochondria becomes highly permeable because of this
change in membrane cytochrome-C escapes in to cytosol.
Cytochrome is an integral part of the oxidative
phosphorylation, electron transport chain and production
of ATP. The escaped cytochrome from mitochondria
causes activation of ‘caspases cascades’. Caspases, this
term represents its function. These molecules contain
cystine at their active site and these molecules have
proteolytic ability as they split a give protein molecule at
a specific position where aspartic acid is located. The
resultant product or residue formed contains four amino-
acids; this group of amino acids is specific to one
caspases. Caspase exists in cytosol or released as
inactivated state called pro-caspases. Caspases behave
like “systematic dismantling device”. When proteins are
chopped in cytosol, the caspase snip the nucleus and its
contents. This step destroys the DNA and shunts its
activity completely. Once DNA is disassembled
structurally and functionally the cytoskeleton of the
apoptotic cell is subjected to “break down”. This whole
chaotic process results in the formation of fine fragments,
these fragments get enclosed in small disposable
membranous pockets called ‘apoptotic bodies’. One of
the important aspects of apoptosis is that the contents of
the dying cell remain enclosed in plasma membrane
through out the entire process. This aspect prevents the
spillage of potential harmful intracellular contents are
avoided but it happens in case of necrosis. The
inflammatory response is also not triggered. Finally the
macrophages engulf the apoptotic bodies thus formed.
Detailed Mechanism
A cell destined to undergo apoptosis has to be guided
by the array of cellular signals. These signals are either
extracellular and/ or intracellular in nature. Popov et al
(2002) and BrSne (2003) have reported on the role of
extrinsic/extracellular signals, also called extracellular
inducers, in the process of apoptosis. The common
extrinsic inducers are toxins, hormones, growth factors,
cytokinins, nitric oxide etc. According to Popov et al
(2002) and BrSne (2003) the extrinsic inducers either
move across the plasma membrane or bring about
variations in the membrane or cell so that the cell is
triggered or induced to receive signals related to apoptosis.
These extrinsic signals are referred as ‘triggers’; these
induce the process of apoptosis in the cell and a molecules
having this ability is termed as positive inducer, this effect
is considered as positive induction. The negative signals
are called repressors as these repose, dampen, inhibit or
puts down the process of apoptosis; these are called
negative inducers and the effect is considered to be
negative induction.
A cell under stress gets initiated to response to those
signals which are intracellular (intrinsic) and induce the
cell to undergo cell suicide. Cotron et al (1998) and Mark
et al (2003) have suggested some of the conditions and
factors which trigger the intrinsic apoptotic signals. These
conditions and factors are, binding of nuclear receptor by
glycocorticoids, heat, radiations, deprivation of nutrients,
viral infection, hypoxia, elevation of Ca++ concentration
 Programmed cell death
within the cell, damage to cell membrane etc. Chiarugi
and Moskowitz (2002) found that many of the cellular
components like ADP ribosome polymerase etc are likely
to help in regulating the process of apoptosis. The extrinsic
and/or intrinsic signals have the ability to activate regulatory
protein system related to initiation of apoptosis. This step
is accomplished before the enzyme system actually starts
the process. If this step is not carried out the target cell
will not undergo apoptosis, thus the above mentioned step
plays a decisive role in the life of cell whether or not to
undergo apoptosis. Basically apoptosis is regulated by
various types of proteins.
Broadly speaking there are two modes of regulating
the apoptosis. First mode is that in which the functioning
of mitochondria is the main target or it is signaled to start
apoptosis and is directly related to its mechanism through
an adopter protein. Second mode involves a step in which
intrinsic inducer is concerned with the increase in the
concentration of Ca++ in the cell; this is achieved by the
drug activity and some of the toxins. Further in this case
“calpain”, (EC3.4.22.52, EC 3.4.22.53, it is a protein which
belongs to the family of calcium dependent, non-lysosomal
cysteine protease) a calcium binding protein, plays key
role.
Mitochondrial Regulatory Mode of Apoptosis
Mitochondria are the site of aerobic respiration and
energy transformation among eukaryotes; any type of
dysfunction of these organelles caused by natural,
metabolic or structural malfunctions stops and/or interrupts
aerobic respiration. This aspect of mitochondria plays a
significant role during apoptotic process. Proteins related
to apoptosis affect mitochondria in different ways such
as increasing the permeability of the membrane of
mitochondria or causing swelling of mitochondria and/or
form pores in the mitochondrial membrane etc. Cotran et
al (1998) observed that such deformities enable the
intrinsic apoptotic effectors to move out of the
mitochondria and initiate apoptosis. Mahon et al (2010)
have found that when permeability of mitochondria
increases enormously then there are more chances of
tumor formation due to intrinsic inducers or pathways than
the extrinsic inducers or pathways. This is due to the
fluctuations in the degree of sensitivity of the mitochondrial
membrane and the cell as a whole one unit. BrSne (2003)
has suggested that there are indications that nitric oxide
have enough ability/potential to squander the membrane
potential of mitochondria and induce apoptosis. As a result
of the changes or increase in permeability of mitochondrial
membrane the specific mitochondrial proteins known as
Small Mitochondrial derived Activator of Caspases
(SMACs), are released in to the cytosol of the cell.
239
Generally these SMACs form a complex with the Inhibitor
of Apoptotic proteins (IAPs) and as a result IAPs are
deactivated, thus IAPs are not allowed to stop or inhibit
the process of apoptosis instead this whole trick favors
the process. Fesik and Shi (2001) have observed that
activity of caspases – a group of cystine protease, is
suppressed by IAPs; this caspase brings about the
degradation of the cell. The degenerative role of a group
of cystine proteases clearly indicates that the permeability
of mitochondria is indirectly regulated by the enzyme
action. Cotran et al (1998) and Laurent (2006) have
reported that outer membrane of mitochondria undergoes
some morphological changes which favor apoptosis, during
these changes a specific channel is formed called
Mitochondrial Apoptosis Induced channel (MAC) and
cytochrome-C is released from the mitochondria through
this channel. Once cytochrome-C reaches cytosol it binds
with Apoptic Activating factor (Apaf-1) and ATP; this
complex gets attached to Pro-caspase–9 resulting in the
formation of a protein complex called Apoptosome. This
Apoptosome is able to convert inactive pro-caspase in to
active form called caspase-9. This active form activates
the effecter caspase-3. Mitochondrial Apoptosis induced
Channel (MAC), also referred as ‘Mitochondrial outer
Membrane Permeabilization Pore’ and various proteins
control its working. Lodish et al (2004) and Dejean et al
(2006) have found that the mammalian gene belonging to
Bcl-2 family of antiapoptotic gene which is homologous
to Ced-9 gene, present in G. elegance, encodes those
proteins which regulate the permeability of mitochondrial
membrane. They further suggested that protein encoded
by Bcl-2 gene can directly affect MAC/MOMPP and
either promote or inhibit apoptosis. Bax and/or Bak help
in the formation of pore in the membrane of mitochondria
while Bcl-2, BCL-XL or Mcl-1 prevents the same.
Extrinsic Inducer Mode of Apoptosis
Currently, direct mode of extrinsic induction of
apoptosis is supported by two mechanisms, at least among
mammals. First mechanism is regarded as Tumor Necrosis
Factor (TNF) induced model and second mechanism is
Fas-Fas-ligand mediated model. Wajant (2002) has
suggested that in both the models intrinsic inducer couples
with a member of family of TNF receptors (TNFRs).
1. Tumor Necrosis Factor Mode (TNF Model)
Cytokines – a small protein molecules secreted by
many cells, these molecules communicate with
intracellular and/or signaling molecules. Macrophages get
activated and produce cytokines; these intracellular
communicators are the major extrinsic mediator of
apoptosis. TNF-R1 and TNF-R2 are the two main
receptors for TNF or most of the cells of human body.
240
Chen and Goeddel (2002) have proposed that when TNF
gets bound to TNF-R1, it initiates the process of activation
of caspases. This initiation is mediated by membrane
proteins which are referred to as TNF-Receptor-
Associated Death Domain (TRADD) and protein called
Fas- Associated Death Domain (FADD). When TNF gets
associated with TNF-R1 there are chances of direct
activation of transcription. This factor is directly related
to the survival of the cell and it is related with inflammatory
response. Specifically during auto-immune diseases there
is a production of abnormal TNF, this step is a key factor
incase of other diseases too.
2. Fax - Receptor Mode of Apoptosis
Fs receptor is also called either Apo-1 or D9s. This
receptor combines with a trans-membrane protein which
belongs to TNF family called Fas-L (Fas-Ligand). Wajant
(2002) has opined that Fas-receptor binds with Fas-Ligand
(Fas-L) and forms a complex called ‘Death Inducing
Signaling complex (DISC). This complex contains FADD,
caspase-8 and caspase-9. He further proposed that cell
may be grouped based on the behavior of caspase or
their mode of action to apoptosis; in one group of cells
that get processed with caspase-8 and has ability to
activate the other member of the family of caspases there
after the apoptosis is carried out. In other group of cells
the Fas-DISC behaves in such a manner so that it helps
the release of pro-apoptotic factors from mitochondria in
large amount and it also increases the activation of
caspase-8 many times.
Common Requirements for Apoptosis
The common components are homodimers, these are
for initiation of apoptotic process among mammals, include
TNF R-1, Fas. These maintain appropriate established
equilibrium between pro-apoptotic members and anti-
apoptotic members. Pro-apoptotic members are Bax, BID,
BAK and/or BAD (death promoting protein) and anti-
apoptotic members are Bcl-XI and Bcl02; these belong
to family of Bcl-2. Murphy et al (2000) have observed
that Bcl-2 members play key role during the drug induced
apoptosis.
Homodimers are concerned with establishing
equilibrium; these components are present on the outer
membrane of mitochondria. The role of pro-apoptotic
homodimers is to make mitochondrial membrane
permeable so that activators of caspases such as
cytochrome-C, SMAC etc, are released from
mitochondria. Generally, during normal conditions anti-
apoptotic members such as Bax or BAk get activated
with BHƒ protein – a member of Bcl-2 family. However,
the regulation of activation or initiation of pro-apoptotic
Y K Lahir
member is still ambiguous incase of non-apoptotic cell
under normal course.
Caspases are highly preserved proteins such as
cysteine dependent aspartate - specific proteases. These
entities create conditions which favour and enforce the
direct apoptotic signals. The caspases are of two types,
namely initiator caspases and effecter caspases; caspases
2, 8,9,10 are initiator caspases while caspases 3, 6, 7 are
effecter caspases. Oligomeric adaptor proteins are
specific adapter proteins which have restricted functional
groups; these oligomeric adapter proteins activate the
initiator caspases by binding with them. These activated
caspases initiate the effectors caspases by bringing
proteolytic cleavage. The active effecter caspases thus
formed react with many of the intracellular proteins and
degrade them by proteolytic activity and ensure apoptotic
process. Susin et al (1999) have observed that apoptosis
can be caused independently of caspases and during this
mode of apoptosis ‘Apoptotic Inducing Factor (AIF)’ is
involved as mediator. Emma et al (2003) have found that
cysteine protease – a member of caspases family play a
vital role in transducing signals which influence the
culmination of apoptosis.
Role of Tumor Suppressor Gene p-53 in
Apoptosis
Stephen et al (1997) have suggested that function of
tumor suppressor gene p-53 is little ambiguous in normal
cell cycle. However when DNA is damaged there is an
increase in the contents of p-53 and cell cycle is either
withheld or delayed till DNA is repaired. In case of degree
of DNA damage is very high the cell is destined to undergo
apoptosis. DNA can be damaged due to very high dose
of UV radiations either solar or other sources. Whenever
the mechanism of p-53 gets impaired because of its
mutation the affected cell with damaged DNA is likely to
survive and it also undergoes cell division causing
chromosomal instability and/or cancer. Pattaway (1998)
while studying on the prognostic markers in clinically
localized prostate gland cancer observed that apoptosis
may play a significant role related to regulating the
functioning of p-53 gene. Generally the apoptosis and p-
53 gene help to regulate cell cycle during developmental
process, progression of prostate cancer. Further, he
proposed that apoptosis ensures damage of the affected
cells. Pattaway (1998) proposed that p-53 puts break to
the cell cycle, initiates cellular repairs and if required then
apoptosis destroys the cell having damaged DNA; in his
opinion there is a need to extend further work on
stimulatory application related to malfunctioning and p-
53. Susan et al (2003) have expressed that when a cell is
subjected to stress, some of the processes and
 Programmed cell death
mechanisms are triggered such as initiation of p-53 tumor
suppressor gene, sequence specific factor for
transcription, induction of arrest of cellular growth,
apoptosis etc. This process of triggering of such processes
and mechanisms depends on factors like type of cells,
type of stress, co-activation of p-53. When stimulation
of p-53 takes place a wide range of net-work of signals
sets in two prime pathways of apoptosis, are triggered
namely, the ‘extrinsic death receptor path way’ and
‘intrinsic mitochondrial path way’. During the process of
triggering of extrinsic receptor path way the “caspases
cascade” is induced or initiated and during intrinsic
mitochondrial path way the equilibrium of Bcl 2 family is
shifted towards pro-apoptotic members; this step promotes
the formation of apoptosome and apoptosis mediated by
caspases. This promotion takes place at a higher rate
when ‘Bid’ is more effective and Bid is target for p-53.The
prime apoptotic effects are triggered because of initiation
of specific apoptotic target genes. Susan et al (2003) are
of the opinion that under certain conditions p-53 has the
ability to enhance apoptosis by carrying out transcription
mechanism independently. Accordingly p-53 employs a
wide range of mechanisms and it ensures effective
initiation of apoptosis at given stage and in a give tissue,
also in a specific manner to cause stress signal. If one
may manipulate the apoptotic functions of p-53, it may
help in cancer therapy.
Role of Rb in Apoptosis
Retinoblastoma – a malignant tumor of the
neurological element of retina occurs exclusively in
children. Stephen et al (1997) have reasoned the
occurrence of retinoblastoma among children and
suggested that it is associated with the role of tumor
repressor gene during development and children having
mutation in retinoblastoma gene Rb. Rb protein is
associated with G1 phase of cell cycle and is responsible
for its completion. Cell enters a phase in which DNA
gets replicated. If Rb protein looses its function, the cell
undergoes uncontrolled cellular proliferation. When a cell
is infected with some DNA tumor virus, this virus is related
to produce a protein which binds with Rb and inactivates
it (the infected cell exhibits similar condition).
Fan and Steer (1999) have worked on the role of
retinoblastoma (Rb) protein in relation to apoptosis; in their
opinion retinoblastoma and its protein (Rb) are related to
the regulation of development, cell growth, differentiation
and cell cycle. Rb is also helpful in protecting cells from
apoptosis. Basically it is believed that Rb is related to
regulating the activities concerned with multiple apoptotic
mediators. The functions of normal cell cycle are
controlled by many factors and Rb is one among them.
241
Rb is quite like nucleophosphoprotein and is needed for a
function of normal cell cycle. When there is damage to
cell cycle or the integrity of DNA in a cell, the concerned
cell is triggered to undergo apoptosis and in this process
Rb is also equally involved. Rb also assists apoptosis path
way along with a transcription factor E‚ F as critical
component either independently or in association with p-
53 tumor suppressor. Fan and Steer (1999) reported that
RB, E‚ F, and p-53 are directly involved in regulating some
of the proteases which play active role in apoptosis. To
summarize, the role of Rb, it can be considered as tumor
suppressor protein which controls cell cycle functions and
also helps to determine the fate of cell through apoptosis.
Vassiliki et al (2005) while studying retinoblastoma
in relation to apoptosis found that Rb cells are resistant to
apoptosis; this resistance is mediated via ‘Death receptor’.
This is because of the deficiency of caspases-8 in these
cells. Probably the expression of caspases-8 is secondary
to epigenetic gene and effectively is reduced due to over
methylation. These researchers are of the opinion that
Rb cell in combination with methylating agents and
modalities which activate death receptor such as TRAIL
receptor and monoclonal anti-bodies are likely to check
retinoblastoma conditions in patients suffering from it. Li
(1999) reported that there are very good chances that
mismatch repair (MMR) also helps in retaining genomic
stability and this is accomplished by stimulating DNA
damage-induced apoptosis by MMR. This occurs as a
cytotoxic response to physical and chemical agent; DNA
adducts along with Mut S/Mut L homologues inducts
apoptosis. This can be done either by provoking repair
event or by blocking steps in DNA metabolism like its
repair and/or replication. MMR proteins recognize the
damage to DNA; this step either stimulates or signals to
activate the cascade for apoptosis. As a result protein
kinase gests activated and it phosphorylate p53 and/or
related p73, these in return initiate the path way of
apoptosis to ensure death of cell to avoid necrosis.
Sequencial Morphological Featrures of an
Apoptotic Cell
There are numbers of signals and path ways which
lead to apoptosis. When a target cell for apoptosis is
triggered after receiving the stimuli, its organelles starts
getting degraded because of the action of activated
proteolytic caspases. During apoptosis cell exhibits some
typical morphological features.
Cell shows shrinkage because the caspases break
down the proteinecious cytoskeleton.
Cytoplasm becomes denser and the cellular parts get
tightly packed.
242 Y K Lahir
Nucleus undergoes ‘pyknosis’. It is a process in which
condensation of chromatin takes place and appears as
compact patches; these patches move towards nuclear
membrane. Pyknosis is the prominent feature of apoptosis.
Santos et al (2000) and (2001) observed pyknosis while
studying apoptosis and referred it to be the “Hall Mark”
of apoptotic cell.
Nagata (2000) has reported that DNA undergoes
“karyorrhexis”. It is a process in which DNA becomes
fragmented within nuclear membrane and/or the
chromatin of the dying cell disintegrates and is distributed
in the cytoplasm. As DNA is undergoing fragmentation
the nuclear envelop also becomes fragmented i.e. it looses
its continuity, as a result the nucleus and DNA splits in to
many discreetly structures called chromatin bodies or
nucleosomes.
Irregular buds like features called “blebs” appear on
the cell membrane.
The cell disintegrate in to small vesicles, these are
called ‘Apoptotic bodies’ which undergo phagocytosis.
The process of splitting and disintegration takes place
quickly and the end products are eliminated vary fast.
Due to this reason it becomes difficult to either visualize
or detect these apoptotic bodies.
Apoptotic cells exhibit “DNA Laddering” phenomenon
while cells dying due to deprivation of nutrition do not.
Miwata et al (1996) suggested that with the help of “test
for DNA laddering “ technique one can differentiate a
cell undergoing apoptosis and the cell undergoing toxic
death (due to deprived nutrition). During apoptosis when
karyorrhexis and activation of endonuclease takes place
the DNA gets fragmented. These fragments get arranged
at regular interval and in size; this arrangement appears
like “ladder” when electrophoresis is done on agar gel.
Clearing of cellular debries formed as a result
of apoptosis
Once the cell has undergone apoptosis cellular debris
are formed. Vandivier et al (2006) has suggested that
neighboring phagocytic cells remove the dead cells and
their fragments or apoptotic bodies as result of
phagocytosis; this process is referred as “efferocytosis”.
Li et al (2003) proposed that phosphotidyl serine is needed
for the clearance of apoptotic cells. Further they have
suggested that cells at final stages of apoptosis, phagocytic
molecules like phosphotidyl serine have been observed
on their cell surface. Wang et al (2003) advocated that
dead cells and/or apoptotic bodies are engulfed; they
suggested that phosphotidyl serine is found on the cytosolic
surface of the plasma membrane but with the help of a
hypothetical protein called “Scramblase”; this is also
redistributed on the outer surface of the apoptotic cell
while Savill et al (2003) are of the opinion that these
molecules (phosphotidyl serine) mark the cell for
phagocytosis. This marking is done by those cells which
have appropriate receptors like macrophages. When the
phagocyte recognizes the apoptic bodies, cell, and its
endoskeleton, engulfment takes place. The removal of
cell debris is carried out in a synchronized manner and no
inflammatory response is elicited to the neighboring cells.
Control of apoptosis
A cell under normal conditions receives “survival
signals” constantly assuring its survival. These signals are
useful to maintain normal internal physiological and
biochemical environment and the ambient external
environment. These survival signals include tissue-specific
growth factors and certain hormones. These signals assist
the cell to maintain appropriate contact with the
neighboring cells and extracellular matrix.
All these survival signals are extra cellular but help
to maintain the survival of the cell and this is achieved as
a result of blockage of ‘caspases cascade’ and prevent
the working of the death mechanism.
Each cell contains in it-self a potent self-destructive
caspases. Under normal conditions, these keep the
powerful enzymes under control; these agents keep the
active enzymes in inactivated form within the cell. This
set up is important to keep the cell alive.
Those cells which are destined to apoptosis have self
destructive caspases. The decision of these cells to
undergo apoptosis or not depends on the presence of
activated caspases.
Those cells which get programmed to die don’t
receive survival signals. In these conditions the lethal
caspases unleashed so to say the growth factors are either
withdrawn or extra cellular matrix is detached to induce
self destruction.
Such programmed cells exhibit “death receptors/death
signals” on/in plasma membrane, e.g., the death receptors
include some “special hormone” or “special chemical
messenger” which are released from WBCs. These death
signals change the path way effectively and may quickly
activate the internal apoptotic mechanism and force the
cell for self destruction. When self-demise-mechanism
gets activated, the specific cell undergoes irreparable
intracellular damage.
Thus, there are some types of signals which promote
apoptosis and some of the signals prevent it. The survival
and death of the cell depends on the competition between
these two types of signals, there is a delicate balance
 Programmed cell death
between these two. This delicate balance may be
destroyed during some diseases which are associated with
the death of cells. Alzheimer’s disease, Parkinson’s
disease, stroke, earlier death of cell which fight infection
during AID infection etc.
Cancer cells do not respond to the extracellular signals
that help the death of cell; these cells do not die or cure
but they grow in uncontrolled manner resulting in the
formation of a mass. This mass is chaotic and uncontrolled.
Apoptosis and its role in immunity
Paul and David (1997) have suggested that apoptosis
plays an important role in the immunological aspects such
as control of immune response, deletion of immune cells,
recognizing self-antigens, cytotoxical killing etc. When
immune system does not function normally then there are
chances of either increase or decrease in the degree of
cell death. There are equal chances of causing spectrum
of autoimmune diseases, immunodeficiency and other
diseases. Vaux and Strasser (1996) opined that apoptosis
is accomplished in two steps, in first step the stimuli trigger
or initiate response in the cell which leads to its death and
this is followed by a mechanism which is responsible for
the implementation of programmed death of cell. Vaux et
al (1994) have proposed that there may be variety of
potential stimuli which initiate the death response in cell
but the mechanism of cell death is same involving
evolutionally conserved effectors having main components
which belong to family of cysteine protease; Alnemri et
al (1996) named these components as “caspases”, further,
once these caspases are activated these components either
directly or indirectly bring about morphological,
biochemical changes and apoptotic changes leading to
the formation of apoptotic bodies. Motoyama et al (1995),
Ito et al (1995), Afford et al (1995), Krams et al (1995)
and Baetze et al (1995) have reported that there are clinical
implications of apoptosis on the immune system and
diseases like hematopoietic defects, allografted kidney and
liver, Chedrak-Higashi syndrome etc. Paul and David
(1997) have mentioned that apoptosis is based on cellular
response with respect to stress, infection, hypoxic
conditions and genetic defects. These researchers also
claim that extensive death on immature lymphocytes being
a normal response to non-productive receptor gene
arrangement. Peter and Denise (2010) are of the view
that primarily the innate immune system rapidly recognizes
the microbes and eliminates them. Detection of microbes
depends on the “germ-line” encoded “pattern recognition
receptors” (PRRs), it indentifies essential bacterial
molecules so called “pathogen associated molecular
pattern” (PAMPs).
Role of apoptosis during development of
243
lymphocytes
Lymphocytes are among the components of blood
and are very closely related to immune system, this
relationship is useful in the functional aspects of
lymphocytes. Paul and David (1997) have suggested that
quite a good numbers of developing T- and B-
lymphocytes are killed during their development. During
this process lymphocytes get differentiated from their
progenitor cells and the related genes present also get
reoriented. The gene orientation assists the reception of
antigens and these antigen receptors are expressed on
their membrane. This recombination exhibits variable
diversity and pattern of joining of segments of these genes;
these joined segments of genes do coding for the receptors,
this process is called V (D) J recombination. Tonegawa
(1983) reported that rearrangement of immunoglobulin
(Ig) gene among B-cells take place independent of the
nature and/or type of antigen, further, there is the formation
of diversity and specificity of immunoglobulin receptor
antigen. He further suggested that the rearrangement of
the receptor gene follow a pattern which is recognized
and this arrangement takes place when these cells pass
through various stages like pro B to pre B1 and pre B-II
stages. When cell is at pro B – stage, it joins DH to JH
regions and later joins the VH to DH gene. During later
stages of development rearrangement of light chain takes
place as cell enters pre B-II stage. It has been observed
that during this development many of the lymphocytes
are dead, the probable reason appears to be the activation
of apoptosis path way. It appears that rearrangement of
gene is favorably transcribed and translated while
receptors are either not expressed on the membrane of
these cells or fail to transduce the signals. Over all these
cells are unable to receive the ‘survival signals’, thus,
subjected to planned cell death because of activation of
apoptotic path way. Once these cells attain a stage in
which Ig genes are oriented favorably and correctly and
Ig is expressed on the cell membrane, there is a cellular
signaling which checks and prevent the signals which
wrongly activate apoptosis.
Godfrey and Zlotnic (1993) reported that progression
of changes in the expression of antigen on surface occurs
during the development of the cells of thymus and markers
such as CD-4 and CD- 8 are secured on the surface of
these cells. Raulet et al (1985) observed that the genetic
orientation of T- cells receptor gene (TCR) occurs in the
thymocytes during their development, this process is
identical to that which takes place in B- cell antigen
receptor genes. Bosma et al (1983), Mombaerts et al
(1992) Shinkai et al (1992 and Blunt et al (1995) have
observed that the specificity of the interaction between
244 Y K Lahir
TCR and MHC or HLA (major histocompatible complex
or human leukocyte antigen), is basically responsible for
the fate, survival or clonal expansion (cell division to
increase in number) or deletion, while cells which are not
able to successfully rearrange their receptor gene are
killed because of their intrinsic apoptotic path way get
activated. These observations were made on phenotype
of severe combined immunodeficiency (SCID) mice and
recombinase activating gene knocks out (rag-1 and rag-
2) mouse stains; circulating receptor-null lymphocytes
were absent in these mice, those developing lymphocytes
which could not produce functional antigens receptors
were also removed. As a result these mice exhibited
hypoplasia of lymphoid organs and mature T- and B-cells
were absent. Observations among ‘scid mice’ revealed
that these mice gave rise to the same phenotype because
their repair of double strand DNA failed during
recombination process which is related to defective DNA
dependent kinase activity. Antigen receptor-Null
lymphocytes were not observed in ‘scid mice’ as these
got killed during their development. The precursors of
lymphocytes were dead due to faulty activation of
apoptosis in these genetically modified or manipulated
(SCID or rag-1) mice resulting in death in large numbers.
Tsujimoto et al (1996) expressed that as other body cells
are regulated by autocrine, paracrine, and endocrine
interaction between different growth factors, the
development and differentiation of immune cells, are no
exception.
The development and regulatory functions of immune
system are governed by a group of interleukins (IL)
belonging to the cytokines family. Interleukins prevent
apoptosis and enhance the chances of survival of immune
cells. Pre B-cell proliferate and survive because of IL-7,
if IL -7 is absent these cells are killed by apoptosis. Spinozzi
et al (1995) observed that IL-4 interleukin is responsible
for the promotion, proliferation, maturation, differentiation
and survival of immature lymphocytes in vivo. Noguchi
et al (1993) found that when interleukins are withdrawn
from those cells which depend on them, these undergo
apoptosis. Further, they found and concluded that IL-2,
IL-4, IL-7 and IL-15 help to receive signals for survival
but under specific condition but when these interleukins
are absent such cells exhibit apoptosis. Paul and David
(1997) reported that glucocorticoids affect lymphocytes.
Most of the cortical thymocytes and precursor stages of
B-cell undergo apoptosis when cortisone is released from
adrenal or subjected to the exposure of dexamethasone.
Further, quite often when lymphoid malignancy is treated
with dexamethasone, these lymphoid cells exhibit
sensitivity to steroids.
Fate of lymphocytes at maturity
During the maturation of lymphocytes attain a stage
at which they express antigen receptors, in the present
context let us consider this stage to be matured and those
which fail to express antigen receptor may be considered
as under developed. In thymus there are two groups of
T-cells with receptor; first group consists of those cells
which fail to interact with self MHC peptides and the
second group of cells exhibit high interaction with MHC
antigen. Nossal (1994) suggested that lymphocytes which
exhibit some type of specific selection based on their
interaction with self antigen, accordingly T-cells having
antigen receptors and do not interact with self MHC
peptides, this feature is considered to be positive selection;
T-cells with receptor antigen and have higher affinity for
self antigen, this feature is considered as negative selection.
Both of these selections are affected by apoptosis. This
role of apoptosis was studied using a special staining
technique called “tunnel staining”, it is a device to detect
the DNA breaks – a characteristics of apoptosis in situ.
Surh and Sprent (1994) reported that in cortical region of
thymus apoptosis was observed in normal mice and also
in those mice which are deficient in MHC; the existence
of apoptosis in indicative of death of majority of cells due
to independent interaction between TCR and MHC
molecules. This affirms that cells which were not positively
selected died due to apoptosis. Miller and Heath (1993)
reported that in case of transgenic mice expressing TCRs
specific self antigens exhibited apoptosis of thymocytes
in the medullary region of thymus, this affirms that
apoptosis is a part of negative selection. They further
observed that dendrictic cells are considered to be cells
with antigen and exhibit negative selection and activate
apoptosis in thymus where T-cells are developing.
Namazee and Burki (1989) suggested that during
development B-cell V (D) J segments get oriented, this
rearrangement results in the expression of IgM and IgD
receptors. Hartley et al (1993) reported that when
experiments were conducted on transgenic mice on the
expression of antibodies towards membrane bound self
antigens, it was observed that apoptosis is the means of
reducing the population of self reactive B-cells.
Regulation of mumber oflymphocytes
(homeostasis of lymphocytes)
Once the lymphocytes become developed in the
primary lymphoid organs these are sent to secondary
lymphoid tissues; these tissues are the sites where
antigenic interaction takes place when lymphocytes are
exposed to an antigen and receive appropriate accessory
signals and the matured B-cells and T-cells get activated.
There after these cells enter cell cycle, increase in number
 Programmed cell death
and get differentiated. After over coming the foreign threat
and their effecter function is accomplished like production
of antibodies (B-cells) or secreting cytokines or kill the
target cells (works as helper and cytotoxic T- cells), the
used up lymphocytes must be removed from the
circulation. When activated cells die the immune response
become restricted and unwanted cells or those cells which
have performed their functions must be killed. Russel
(1995) proposed that there is a chance that some of the
cells develop the ability to recognize and produce a
response to self antigen and some of them may skip
selection within thymus. Alderson et al (1995) observed
that mostly activation induced cell death (AICD) is
probably connected with induction on cells inducing CD-
95 (Fas/AOP-1) and its ligand; this is likely to induce
suicide or fratricide within the activated cells. In addition
to these factors other TNF receptor member like TNF
R-2, p-75 and APO-2 have also been observed to be
involved in activation induced cell death (AICD). Thus,
one can observe the role played by apoptosis in activating
lymphocytes and also in maintaining optimum numbers of
lymphocytes so to say apoptosis is essential in retaining
specific tissue-turn over in the primary and secondary
lymphoid tissues. Fraker and Lill (2004) have reported on
the impact of aging on over all effect of apoptosis; as an
individual becomes old the rate of production of native T-
cell and B-cell and their killing capacity reduces.
Affinity for maturation
Shlomchik et al (1987), Liu et al (1989), Pulendran et
al (1995) and Shokat et al (1995) have opined that the V-
region of Ig gene subjected to somatic hyper mutation
after activation within the germinal loci of lymphoid tissue
and this brings about the formation of antibodies having
elevated tendency to interact with antigen. Apoptosis
efficiently removes those cells which do not develop
antibody with higher affinity and even those cells which
have mutation permitting the antibody to interact with self
antigens. This whole process is referred as “affinity for
maturation.
Apoptosis and cytotoxicity
Paul and David (1997) explained the process of
cytotoxic killing cells involving apoptosis. According to
them cell mediated cytotoxicity is a tendency found in
“specific immune cells” and “natural killer cells”, these
cells either destruct or kill the target cell but themselves
do not loose their integrity. Such cells mediated immune
cells are cytotoxic T-lymphocytes (CTL) and “natural killer
cells”. The target cell is identified due to the interaction
within the receptors present on the killer cells (TCR) and
antigen present on the surface of target cell. The
destruction of target cell is attained by two modes, first
245
mode involve exocytosis. Exocytosis is accompanied by
the specific granules containing “perforin” and
“granzymes” (serine proteases), the second mode
involves CD-95-ligand.
Shiver et al (1992) proposed that perforin is a specific
protein which has the ability to punch whole on the surface
of the target cell while granzymes (serine proteases) are
the enzyme like molecules which can induce DNA
degradation in target cell. Generally target cells are viral
infected cells; such target cells are identified by interaction
between TCR (receptors on the killer cell) and those on
the surface of cytotoxic T lymphocyte - CTL. Shresta et
al (1995) suggested that after this interaction
degranulation takes place, this step permits perforin to
make wholes on the surface of target cell, through these
wholes granzymes molecules enter the target cell and
degrade the DNA of the target cell. Darmon et al (1995)
observed that these steps cause cleavage and activation
of apoptotic cysteine proteases (PPZC). Rouvier et al
(1993) demonstrated in vitro, that CD-95 is related in
accomplishing cytotoxicity but its intensity is relatively quite
low in comparison to the CTL killing induced by perforin
and granzymes.
Clinical involvement of apoptosis in immune
reaction
Paul and David (1997) have discussed the probable
correlation between disease and apoptosis; they proposed
that either change in cell are caused indirectly or due to
secondary effectors of disease or disease likely to be the
direct effect because of the mechanism of apoptosis. They
argued that mostly apoptosis is a defensive response when
a cell is infected by pathogen or experiences stress, hypoxic
conditions, genetic artifact; there are every chance of
cell may die because of apoptosis.
Afford et al (1995), Ito et al (1995) and Krams et al
(1995) have discussed the clinical involvement of apoptosis
under different diseased conditions. Normally immature
lymphocytes are killed in large number during non-
productive receptor gene orientation, when transplant is
rejected by the patient, apoptosis induced by CTL on graft
cell, immunodeficiency has been observed during Chediak
– Higashi syndrome (immune system related genetic
defect), state of inability of CTL of destroying target cells
by degranulation, genetic defect in CD-95 gene causes
autoimmune lymphoproliferation disorder. Emma et al
(2003) have found that cysteine proteases - members of
caspase family play a vital role in transducing signals
which influence the culmination of apoptosis; they
proposed that caspase-1 have its role during signaling
immune response against microbial pathogen, it gets
associated with cytokines like interleukin 1-B (IL-1B),
246 Y K Lahir
IL-18, and help in functioning of pro-inflammatory
cytokines. They are of the opinion that caspases get
activated during apoptosis and inflammation there by
assisting both these processes. Fraker and Lill – EL (2004)
studied the effects of aging and nutritional status of an
individual, according to their studies as age of an individual
increases the immune system becomes less potent and
dysregulated affecting the functional aspects of apoptosis.
The common dysregularities caused are reduced
production of native T- and B- cells and their cytolytic
killing capacity, accumulation of large number of malignant
cells, increased inflammatory response etc. Study on the
impact of nutritional status on apoptosis revealed that
deficient nutrition and individual nutrient can bring defective
abnormalities in the functioning of apoptosis. A small
duration of zinc deficiency caused 50% to 300% of
apoptosis among pre B- and pre T- cells resulting in
elevated lymphopenia and atrophy of thymus. Zinc
deficiency resulted in 80% reduction in pre B-cells in 28
months old mice. Fraker and Lill –EL (2004) have
discussed that probably zinc is released form neurons
during Alzheimer, Parkinson’s disease and brain injury and
this causes accelerated apoptosis and damage to the
ambient tissue. The ill effects of free zinc bring about
apoptosis of higher derogative potential in the variety of
tissues. They further suggested that marked contrast to
sub-optimal zinc, there are chances of initiation of caloric
restriction in young mice and under these conditions
autoimmunity and immunosenescence are delayed. Peter
and Denise (2010) have studied the molecular mechanism
of inflammosome (a pathogen and danger sensing complex
triggering innate immunity) activation during the microbial
infection; they proposed that the innate immune system
is very helpful in rapid recognition and to eliminate
infectious microbes. The process of detection of microbes
depends on the “pattern recognition receptors” (PRRs),
this pattern helps to identify the molecular aspects of
bacteria, this identification is named as “pathogen-
associated molecular pattern” (PAMPs). A sub-
component of PRR and belonging to “NOD- like receptor”
(NLR) and PYHIN are specific protein group, these help
to detect pathogenic virus and bacteria which are present
in the cytosol of the host cells. These also induce a group
of multiprotein signaling device called “inflammosome”.
This device assists in activation of central mediation of
innate immunity of mammalian cysteine protease –
caspase – 1. When caspase-1 gets activated it helps the
pathway involved in maturation and release of interleukin
1- β (IL-18) and proteins which protect and repair the
cell and tissues. Caspase-1 performs an additional function
in which it initiates a different form of death of cell called
“PYROPTOSIS”. Thus inflammosome adaptor ASC in
caspase-1 and autoproteolysis are helping microbial
infection such as Franscisella tulatinsis and Salmonella
typhimurium. Indran et al (2011) have observed that
defective and inefficient apoptosis is an acquired feature
of cancer cells. A detailed understanding of signaling
mechanism involved in apoptosis and the resistant
mechanism of apoptosis are likely to lead to effective
and target oriented therapeutic strategies to tackle cancer.
REFERENCES
Afford S C, Hubscher S, Strain A J, Adams D H and Neuberger J M
(1995) Apoptosis in human liver during allograft rejection and
end stage liver disease. J. Pathol. 176, 373-380.
Albert Keith, Johnson Alexander, Lewis Julian Raff, Martin Roberts,
and Walter Peters (2008) Apoptosis: programmed cell death
eliminates unwanted cells. Molecular Biology of Cell: 5th Ed,
Garland Science, Pp 1115.
Alderson M R, Tough T W andDavis-Smith T (1995) Fas ligand
mediates activation-induced cell death in human. J. Exp. Med.
181, 71-77.
Alnemri E S, Livingstone D J and Nicholson D W (1996) Human ICE/
CED-3 protease nomenclature. Cell 87, 171.
Baetz K, Isaaz S and Griggiths G M (1995) Loss of cytotoxic T-
lymphocyte function in Chediak-Higashi syndrome arises from
a secretory defect that prevents lytic granules exocytosis.
J. Immunol. 154, 6122-6133.
Blunt T, Finnie N J and Taccroli G E (1995) Defective DNA dependent
protein kinase activity is linked to V (D) J combination and
DNA repair defects associated with the murine scid mutation.
Cell 80, 813-823.
Bosma G C, Custer R P and Bosma M J (1983)A severe combined
immunodeficiency mutation in the Mouse. Nature 301,527-530.
BrSne B (2003) Nitrogen oxide: No apoptosis or turning it on? Cell
Death Differ. 10, 864-869.
Chiarugi A and Moskowitz M A (2002) PARP-a preparatory of
apoptotic cell death? Science 297, 259-263.
Cotran Kumar and Collins (1998) Robbins Pathologic bases of diseases.
SW Saunders Co Philadelphia.
Darmon A J, Nicholson D W and Bleakley R C (1995) Activation of
apoptotic protease CP32 by cytotoxic T-cells derived granzyme
–B. Nature 377, 446-448.
Dejean L M, Martinez-Caballeros, Mohan S and Kinnally K W (2006)
Regulation of mitochondrial apoptosis – induced channel (MAC)
by Bcl-2 family protein. Biochem. Biophys. Acta 176, 191-201.
Emma M C, Helen C and Seamus J M (2003) Caspase-activation
pathway in apoptosis and immunity. Immunol. Rev. 193, 10-
23.
Fan G and Steer C J (1999) Role of retinoblastoma protein in apoptosis,
Apoptosis 4, 21-29.
Fesik S and Shi Y (2001) Controlling Caspases.Science 294, 1477-
1478.
Fraker P J and Lill-EL Ghanian D A (2004) Many roles of apoptosis
in immunity as modified by Aging and nutritional status. J.
Nutri. Health Aging 8, 63.
Godfrey D I and Zlotnik A (1993) Control points in early T-cell
development. Immunol. Today 14, 547-553.
 Programmed cell death
Hartley S B, Cooke M P and Fulcher (1993) Elimination of self-
reactive B-lymphocytes proceeds in two stages: arrested
development and cell death. Cell 72, 325-335.
Indran I R, Gregory T, Shazib P and Catherine B (2011) Recent
advances in apoptosis, Mitochondria and drug resistance in
cancer. Biochim et Biophysica Acta-Bioenerg. 1807, 735-745.
Ito H, Kasagi N, Shoman K, Osaki M and Adachi H (1995) Apoptosis
in human allografted kidney. Analysis by terminal
deoxynucleotidyl transferase mediated DUTP –biotin nick end
labeling. Transplantation 60, 794-780.
Krams S M, Egawa H and Quinn M B (1995) Apoptosis as a
mechanism of cell death in liver allograft rejection.
Transplantation 59, 621-625.
Laurent M D, Sonia M and Kinnally K W (2006) Is MAC the knife to
cut the cytochrome-C from mitochondria during apoptosis? Cell
Death and Differentiation 13, 1385-1387.
Li G M (1999) The role of mismatch repair in DNA damage induced
apoptosis. Oncol. Res. 11, 393-400.
Li M O, Sarkisian M R, Mehal W Z, Racik P and Flavell R A (2003)
Phosphatidylserine is required for the clearance of apoptic cells.
Science 302, 1560-1562.
Liu Y J, Joshua D E and Williams G T (1989) Mechanism of antigen-
derived selection in germinal centers. Nature 342, 929-931.
Lodish Harvey et al (2004) Molecular cell Biology. W H Freeman and
Co. New York.
Mohan S, Bustamam M A, Abdelwahab S I, Alzubairi A S, Aspollah
M, Abdullah R, Ehassan M M, Ibrahim M Y and Syam S (2010)
Typhonium flabelliform induces apoptosis in CEMss cells via
activation of caspase-9, PARP cleavage and cytochrome-C release
in its activation coupled with GO/GI phase of cell arrest. J.
Ethnopharmacolgy 127, 112-117.
Mark P, Mattson and Sic L Chau. (2003) Calcium orchestrates
apoptosis. Nature Cell Biology 5, 1041-1043.
Miller J F and Heath W R (1993) Self ignorance in the peripheral T-
cell pool. Immunol. Rev. 133, 131-150.
Miwata D, Mayerson B, Torok-strob and Zagor A (1996) An
evaluation of renal tubular DNA laddering in response to O‚
deprivation and O‚ injury. Retrieved (2006) -04-07.
Mombaerts P., Iacomini J and Johnson R S (1992) Rag-1-deficient
mice have no mature B and T- lymphocytes. Cell 68, 869-877.
Motoyama N, Wang F and Roth (1995) Massive cell death of immune
hematopoietic cells and neurons in Bcl-x-deficient mice. Science
267, 1506-1510.
Murphy K M, Ranganathan V, Farnsworth M L, Kavallaris M and
Lock R B (2000) Bcl-2 inhibit Bax translocation from cytosol to
mitochondria during drug induced apoptosis of human tumor
cells. Cell Death Differentiation 7, 102-111.
Nagata S (2000) Apoptic DNA fragmentation. Exp. Cell Res. 256, 12-
18.
Namazzee D A and Burki K (1989) Clonal deletion of B-lymphocytes
in a transgenic mouse bearing anti MHC lass antibody gene.
Nature 337, 562-566.
Nichola S Mitsiades (2005) Human retinoblastoma cells are resistant
to apoptosis induced by Death receptors: role of caspase-8 gene
silencing. Invest. Opthalmol. Vis. Sci. 46, 358-366.
Noguchi M, Yi H and Rosenblatt H M (1993) Interleukin-2 receptor
gamma chain mutanon results in x-linked server combined immune
247
deficiency in human. Cell 73, 147-157.
Nossal G J (1994) Negative selection of lymphocytes. Cell 76, 229-
239.
Pattaway C A (1998) Prognostic markers in clinically localized prostate
cancer. Tech. Urol. 4, 35-42.
Paul G Ekert and David L Vaux (1997) Apoptosis and the immune
system. Brit. Med. Bull. 53, 591-603.
Peter Broz and Denise M Monak (2010) Molecular mechanism of
inflammosome activation during microbial infection. Immunol.
Rev. 243, 174-190.
Popov S G, Villas Mill R and Bernardi J (2002) Lethal toxin of Bacillus
anthracis causes apoptosis of macrophages. Biochem. Biophys.
Res. Comm. 293, 349-355.
Pulendran B, Kannourakis G, Nouri S, Smith K G and Nossal G J
(1995) Soluble antigen can cause enhanced apoptosis of germinal
center B-cells. Nature 375, 331-334.
Raulet D H, Garman R D, Saito H and Tonegawa S (1985) Development
regulation of T-cell receptor gene expression. Nature 314, 103-
107.
Rouvier E, Luciant M F and Goldstem P (1993) Fas involvement in
Caz z† T- cell mediated cytotoxicity. J. Exp. Med. 177, 195-200.
Russell J H (1995) Activation-induced death of mature T cells in
regulation of immune response. Curr. Opin. Immunol. 7, 382-
388.
Santos A, Susin D E, Ravagan L, Samejimes K, Zimzami N, Leoffler
M, Cosntantini P and Ferri K F (2000) Two distinct pathways
leading to nuclear apoptosis. J. Exp. Med. 192, 571-580.
Santos A, Madelein K, Imreh G and Hallberg E (2001) Sequential
degradation of protein from nuclear envelop during apoptosis.
J. Cell Sci. 114, 3643-3653.
Savill J, George C and Hasett C (2003) Eat me or Die. Science 302,
1516-1517.
Shinkai Y, Rathbun G and Lam K P (1992) RAG-2- deficient mice
lack mature lymphocytes owing to inability to initiate V (D) J
rearrangement. Cell 68, 855-867.
Shiver J W, Su L and Henkart P A (1992) Cytotoxicity with target
DNA break down by rat basophilic leukemia cells expressing
both cytolysin and granzyme A. Cell 71, 315-322.
Shlomchik M J, Marsh K, Rothstem A, Wolfowieze C B, Rothstem
T L and Weigert M G (1987) The role of clonal selection and
somatic mutation in autoimmunity. Nature 328, 805-811.
Shokat K M and Goodnow C C (1995) Antigen induced B-cell death
and elimination during germinal center immune responses. Nature
375, 334-338.
Shresta S. Mavlover D M, Hensel J W, Russel J H and Ley T J (1995)
Natural killer and Lymphokine activated killer cells require
granzyme B for the rapid induction of apoptosis in susceptible
target cells. Proc. Nat. Acad. Sci. U S A 92, 5679-5683.
Spinozzi F, Nicoletti I and Agea E (1995) IL-4 is able to reverse the
CD-4, CD-6 alpha, beta and/or gamma, delta T lymphocytes.
Immunol. 86, 379-384.
Stephen R B, Jeremy S H, Elizabeth A S and Huge A W (1997) From
genes to cell. A John Wiley and Sons publication, New York.
Surh C D and Sprent (1994) T- cells apoptosis detected in situ during
positive and negative selection in thymus. Nature 372, 100-103.
Susan H, Michael B, Zehavit G and Ygal H (2003) Apoptosis- the p-
 248
53 net work. J Cell. Sci. 116, 4077-4085.
Susin S A, Lorenzo H K and Zamzami N (1999) Molecular
characterization of mitochondrial apoptosis–inducing–factor.
Nature 377, 441-446.
Tannapfel A, Weinans L, Geissler F, SchStz A, Katalinic F, Hanss T
and Writtekind C (2000) Mutations of p-53 tumor suppressor
gene apoptosis and proliferation in intrahepatic cholangiocelluar
carcinoma of liver. Diab. Dis. Sci. 45, 317-324.
Tonegawa S (1983) Somatic generation of antibody diversity. Nature
302, 575-581.
Tsujimoto T, Lisukov I A, Huang N, Momoud M S and Kawano M M
(1996) Plasma cells induce apoptosis of pre B cells by interacting
with bone marrow stomal cells. Blood 87, 3375-3383.
Vandivier R W, Hanson P M and Douglas I S (2006) Burying the dead:
the impact of failed apoptic cell removal (efferocytosis) on
View publication stats
Y K Lahir
chronic inflammatory lung disease. Chest 129, 1673-1682.
Vassiliki P, Constantine S M, Cinaran M, Galionos F J Negri, Athina
G, Loannis X, Geraassimos V, Sophia T, Joan WM and Nicolas
M (2005) Human retinoblastosoma cells are resistant to
apoptosis induced by death receptors: role of caspase-8-gene
silencing. Invest. Opthalmol. Vis. Sci. 46, 358-366.
Vaux D L and Strasser A (1996) The molecular biology of apoptosis.
Proc. Nat. Acad. Sci. USA 93, 2239-2244.
Vaux D L, Haecker G and Strasser A (1994) An evolutionary perspective
on apoptosis. Cell 76, 777-779.
Wajant H (2002) The Fas signaling pathway – more than a paradigm?
Science 296, 1635-1636.
Wang X, Wu Y C, Fadok V A, Lee M C, Gengyo A, Cheng L L,
Ledwich D and Hsu P K (2003) Cell corpse engulfment indicated
by C. elegance, Phosphatidylserine receptor through CED 5
and CE-12. Science 302, 1563-1566.