Leishmania donovani Inhibits Apoptosis of Infected Macrophage

Poster by Lisa Fong, Research by Kathryn J. Moore and Greg Matlashewski

Gel 1 Gel 2
Introduction
Leishmania donovani is a flagellated protist that causes kala-azar or visceral
leishmaniasis, an often fatal disease with symptoms of fever and hepatic/splenic
swelling. L. donovani is spread from an infected rodent, canine, or human host to a
healthy one by the bite of a sand fly. When introduced to a new host, the parasite
waits to be engulfed by a macrophage, where it begins to multiply by binary fission.

The colony-stimulating cytokine M-CSF is required to sustain the viability of a

macrophage culture. In its absence, mass occurrences of apoptosis or programmed
cell death is triggered, resulting in fragmentation of DNA and membrane blebbing.
Figure 2. Although infection by L. donovani induces cytokine gene expression of TNF- α,
The immune system utilizes this process to destroy infected macrophages and any
GM-CSF, TGF-α, and IL-6, of those only TNF-α and GM-CSF (lane 4, 5 of Gel 1) inhibited
parasites harbored inside them. Key to L. donovani’s virulence is its ability to
apoptosis in BMMs.
counter the apoptotic process and thus evade eradication attempts by the host
immune system. This experiment investigated the effect of L. donovani infection on
preventing macrophage apoptosis in the absence of M-CSF and the mechanisms by
which it may do so.

Results
Tests were conducted using 5-day-old rodent bone marrow-derived macrophages
(BMMs). PCR and electrophoresis using a 1% agarose gel were used to visualize DNA
from macrophages under different conditions to determine whether apoptosis had
occurred or not. Each test was conducted at least three times. Figure 3. LPG, a common molecule on L. donovani’s surface, also inhibits apoptosis of
BMMs at sufficient concentrations.

Conclusions
• As a parasite that requires host macrophages for the purpose of reproduction, L. donovani is
able to prevent infected macrophages from undergoing apoptosis.
• L. donovani inhibits apoptosis in macrophages deprived of M-CSF by inducing expression of
TNF-α and GM-CSF or exposing macrophages to LPG, in effect replacing M-CSF with other
survival factors.

Figure 1. Infection by Leishmania donovani inhibits apoptosis of macrophage. DNA

Reference
was isolated from BMMs after undergoing the following treatments: lane 1, BMMs Moore, Kathryn J., and Greg Matlashewski. "Intracellular Infection by Leishmania donovani
grown in FCS (fetal calf serum) containing 10% M-CSF; lane 2, BMMs isolated Inhibits Macrophage Apoptosis." Journal of Immunology 152.6 (1994): 2930-937. American
without FCS or M-CSF for 24 hours; lane 3, BMMs grown in FCS without M-CSF; lane Association of Immunologists. 1 Nov. 2010. Web.
4, BMMs infected with L. donovani for 24 hours. Data shows that at 10% or lower
M-CSF, apoptosis occurs in a macrophage population but does not occur when Willey, Joanne, Linda Sherwood, and Christopher Woolverton. Prescott, Harley, & Klein's
macrophages are infected by L. donovani. Microbiology. 7th ed. New York: McGraw-Hill, 2008. 1004-1006. Print.