Life Cycle of Leishmania

Life Cycle of Leishmania

Leishmania species all follow the same general life cycle pattern (Figure 1).
The parasite has two main morphological forms, termed ‘amastigotes’ and
‘promastigotes’, which are found in the mammalian and sandfly hosts
respectively. Amastigotes are amongst the smallest of eukaryotic cells, being
ovoid cells of 3–5 mm on the mainaxis. They possess a nonfunctional flagellum,
which does not project beyond the cell body, and are not capable of movement.
The base of the flagellum originates in the flagellar pocket, a specialized
infolding of the surface membrane. The nucleus is centrally located and adjacent
to the smaller kinetoplast. About 10% of the total cellular DNA is found in the
kinetoplast, the remainder in the nucleus. Promastigotes are larger and
elongated, with a cell body length of 5–15 mm. The flagellum is functional in
promastigotes, projecting from the flagellar pocket to extend beyond the cell
body, and these are active motile forms. The flagellum is at the anterior end of
the cell, its motion pulling the cell body along behind.
Amastigotes are largely intracellular stages that live in the phagolysosomal
system of macrophages. No other host cell type is used. However, macrophages
are themselves a heterogeneous group and different subpopulations are involved
in different types of clinical disease. For example, skin macrophages are
parasitized in the case of cutaneous leishmaniasis, whereas Kupffer cells in the
liver are parasitized in visceral disease. Macrophages are phagocytic and readily
engulf amastigotes, followed by phagosome– lysosome fusion. This microbial
defence mechanism is lethal to most foreign organisms, as it results in exposure
to an acidic pH of 4.5–5.5 and attack by a battery of lysosomal enzymes.
However, Leishmania can survive this experience, and indeed thrive in this
environment. Within the phagolysosomal environment the parasites grow and
divide, such that an individual macrophage may eventually contain many tens of
amastigotes. There is no specific escape mechanism known, and probably in
most cases the host cell simply ruptures when it cannot accommodate any more
parasites, which are then taken up by further macrophages
The mammalian phase of the life cycle is chronic, overt signs of disease
lasting from months to a lifetime. Even in human cases where clinical cure is
achieved, either naturally through an immune response or by chemotherapy, it
is doubtful that the parasite is completely eliminated. If a patient is subsequently
immunocompromised, for eg human immunodeficiency virus (HIV) infection, this
can lead to a reactivation of disease. The same applies to canine leishmaniasis,
which is more difficult to treat than human infection, and where parasites are
also likely to persist even if clinical cure can be achieved. Wild mammals infected
with Leishmania almost certainly remain so for life, usually with little overt sign
of disease, and provide a long-term source of infection for humans.
Transmission of disease is by female sandflies, which are widely distributed
through tropical, subtropical to temperate regions of the world (Lane, 1993). Of
the 700 or so described species of phlebotomine sandfly, approximately 70 have
been implicated in transmission of leishmaniasis. Female sandflies require a
bloodmeal to provide nutrients for egg production. The females are pool-feeders,
that is they use their mouthparts to probe and slice through the surface
capillaries of the skin, then imbibe blood from the wound. So, despite their small
size of 2–4 mm, the bite of a sandfly can be painful. Female sandflies acquire
the parasite if, during the course of feeding on an infected mammalian host,

©Uttam Kr. Patra. 2007 1

 Life Cycle of Leishmania

Leishmania-infected macrophages and/ or free amastigotes are liberated into the

wound site. These are then taken up by the sandfly along with the bloodmeal.

Figure 1. Leishmania life cycle in mammalian hosts (upper) and in sandfly vectors (lower).
Mammalian stage: (1) delivery of infective promastigotes into mammalian skin by the bite of sandfly vector;
(2) phagocytosis of promastigotes by a macrophage; (3) fusion of the phagosome containing a promastigote
with the lysosome; (4) differentiation of promastigote into amastigote in the phagolysosome of the infected
macrophage; (5) replication of an amastigote in a parasite-containing or parasitophorous vacuole; (6)
formation of large parasitophorous vacuole and continuing replication of intravacuolar amastigotes; (7) rupture
of heavily parasitized macrophage and release of
amastigotes; (8) phagocytosis of released amastigotes by a macrophage.

Sandfly stage: (9) ingestion of parasitized macrophages by sandflies after a blood meal taken from infected
humans or reservoir animals; (10) rupture of the ingested macrophages and release of amastigotes in the gut
of sandflies; (11) replication of amastigotes and their differentiation into promastigotes; (12) replication of
promastigotes in midgut and insertion of their flagella into microvilli of the gut epithelial cells; (13) replication
of L. brasiliensis group in the pylorus and ileum of the sandfly hindgut as paramastigotes with broadened
flagella attached to the chitinous gut wall via hemi-desmosomes; (14) forward movement of promastigotes to
thoracic midgut as haptomonads with broad flagella attached to the chitinous gut wall; (15) sessile
paramastigotes with broad flagella attached to the chitinous wall of stomadeal valve, pharynx and buccal cavity
or cibarium; (16) actively motile and infective promastigotes found in the proboscis, or mouth part, of
sandflies.

©Uttam Kr. Patra. 2007 2

 Life Cycle of Leishmania

The bloodmeal of a few microlitres in volume accumulates in the abdominal

midgut of the sandfly and the digestive process is initiated. The bloodmeal is
enclosed in a peritrophic matrix, a lattice-like network composed of chitin fibrils
and glycoproteins, which is secreted by the gut epithelial cells. These also
secrete digestive enzymes to enable breakdown of the bloodmeal nutrients, of
which haemoglobin is a major component. Amastigotes are liberated from their
degrading macrophage host cells and become extracellular within the bloodmeal.
They rapidly differentiate into promastigote forms by elongation of their cell
body and extension of the flagellum, and begin to multiply within the bloodmeal.
Like their amastigote counterparts, the promastigotes are well adapted to
survive in this hydrolytic environment and can withstand exposure to the sandfly
digestive enzymes. The promastigotes grow, divide and increase in numbers,
initially within the confines of the bloodmeal contained in the peritrophic matrix.
After 3–4 days the peritrophic matrix begins to break down, assisted by the
action of a parasite secretory chitinase. This enables the promastigotes to
escape the abdominal midgut and become fully established in the vector. Those
in the subgenus Leishmania do so by spreading forward, attaching to the
microvillar border of the midgut epithelium, to the cuticular surface of the
stomodeal valve and foregut, and by embedding themselves in a gel-like matrix
of parasite origin in the anterior midgut. Those in the subgenus Viannia do so by
migrating backwards into the hindgut and attaching to the cuticular surface
found there. They multiply at this site for a period and then migrate forward to
the anterior midgut. Eventually, in both subgenera, promastigotes are found in
large numbers in the anterior midgut and foregut 1– 2 weeks after the initial
bloodmeal. From this position they can be transmitted when the sandfly takes
her next bloodmeal.
Transmission is effected by a specific life-cycle stage, the metacyclic
promastigote. During their multiplication and migration in the sandfly gut, the
promastigotes undergo various morphological and biochemical changes.
Metacyclic promastigotes are produced towards the end of the developmental
sequence and possess properties that preadapt them for survival in a
mammalian host. They are biochemically intermediate between other
promastigotes and amastigotes, expressing some genes in common with
promastigotes, others in common with amastigotes, as well as metacyclic-
specific genes. They are also highly infectious to a suitable mammalian host in
comparison with other promastigote forms, as a result of their ability to avoid
complement-mediated lysis and to stimulate their own uptake by macrophages.
Metacyclic promastigotes are dedicated to transmission and represent a
significant proportion of the total promastigote population in a mature infection.
When the infected sandfly probes and feeds on a new mammalian host,
metacyclic promastigotes are inoculated into the wound via the proboscis.
Interestingly, infected sandflies experience more difficulty in engorging and
probe more frequently than uninfected flies, which enhance parasite
transmission. This appears to be due to the solid mass of parasites embedded in
their gel-like matrix in the anterior midgut. Once deposited in the skin,
metacyclic promastigotes are engulfed by macrophages, differentiate into
amastigote forms, and take up their intracellular residence.

©Uttam Kr. Patra. 2007 3