AP 305: HONEYBEE PESTS

AND DISEASES

INNATE IMMUNITY OF HONEYBEE TO INFECTION

 Introduction
• Insects colonize a wide range of ecosystems in which
they are exposed to a wide range of infectious
microorganisms.
• Honey bees are exposed to a wide range of infectious
microorganisms, including Gram-positive bacteria
(example?), Gram-negative bacteria (example?),
protozoa, fungi and viruses.
• Ectoparasitism by other arthropods increase infectious
risks due to increased exposure to infections and
alteration of honey bee immunity.
• Economic benefits of certain insects including honey bee
lead to studies of resistance of insects to infections.
 Innate Immunity
• Innate Immunity is the first line of host defense that
allow not only animals but also plants to detect infections
and to activate a set of responses to control the infection.
• Even the more advanced Adaptive immune response in
vertebrates is triggered and shaped by a dedicated set of
cytokines and co-stimulatory molecules produced by the
innate immune system.
• Studies of insect immunity reveal that molecular signaling
cascades involved in host defense have been conserved
through evolution.
• Apis mellifera genome contain orthologues for most
immune related genes identified in other species.
 …Innate Immunity…
• Insect host-defense system involves both humoral and
cellular aspects.
• Cellular response constitute of Phagocytosis by special
blood cells, Melanization and Encapsulation of large
invaders.
• Humoral responses involve chains of events that begin in
the hemolymph – an insect correlate of blood
• Circulatory and membrane associated receptors
recognize molecular structures (PAMPs) from invading
microorganisms.
• Response of insects vary depending on challenges by
different types of infectious microorganisms
 Humoral Response to Bacteria and Fungi
• Humoral immunity is mediated by macromolecules found in
extracellular fluids.
• The fat body in insects plays a very important role in response
against infections.
• Upon infection, by bacteria or fungi, the fat body is induced to
synthesize and secrete a cocktail of potent cationic
antimicrobial peptides into the hemolymph.
• In addition, a number of other molecules are involved in host
response against infections.
• These peptides are also expressed in hemocytes and surface
epithelia either constitutively or upon infection.
• Secretion of AMPs is a conserved feature of innate immune
responses in vertebrates and invertebrates.
 Major molecules secreted to help control infections in insects
SECRETED MOLECULES FUNCTION

Antimicrobial peptides Lysis/Disrupting the structure or function of

microbial cell membranes/target specific
microbial macromolecules
Nitric oxide synthase Toxic against pathogens/Regulates the functional
activity, growth and death of immune system
(synthesize NO) cells.
Phenoloxidase Melanization/Produce a set of intermediate
products such as quinones, diphenols,
superoxide, hydrogen peroxide, and reactive
nitrogen intermediates
Thioester containing proteins Covalently binds to the surface of invading
pathogens, promoting phagocytosis and
(TEP1) lysis/Act as a pattern recognition receptor (PRR)
 Humoral Response to Bacteria and
Fungi
• AMPs can be categorized into three groups depending on
their main microbial targets

• Honey bees respond to infection by producing a number

of antimicrobial peptides which share properties with
their Drosophila counter parts

• Properties of AMPs include:

– small size,
– cationic nature,
– broad activity spectrum and modes of action (see table below)
 Humoral Response to Bacteria and
Fungi
Activity against… AMPs in Drosophila AMPs in Honeybees

Gram negative Diptericin Hymenoptaecin

bacteria Drosocin Abaecin
Attacin Apidaecin
Cecropin

Gram positive Defensin Defensin-1

bacteria Defensin-2

Fungi Metchnikowin None

Drosomycin
 Cellular Immunity
• Cellular immunity in honeybees involves
specialized types of blood cells.
• Cells found in hemolymph of honeybees can be
grouped into different classes depending on their
morphology and function.
Plasmatocytes

Hemocytes Crystal cells

Lamellocytes
 Cellular Immunity
• Plasmatocytes are the major cell type in the
circulation and within tissues.
• They are dedicated phagocytes that resemble
mammalian monocytes/macrophages.
• They patrol tissues and help clear apoptotic
corpses and foreign microbes.
• They also signal infection to the fat body.
• Plasmatocytes (phagocytes) express phagocytic
receptors for phagocytosis of apoptotic cells.
 Cellular Immunity
• Crystal cells represent only about 5% of the blood cells
in insects.
• Contain crystalline inclusions including enzymes
involved in humoral melanization - phenoloxidase.
• PO is secreted by crystal cells and activated by a serine
protease PAE (phenoloxidase activating enzyme).
• PO catalyze oxidation of mono- and diphenols to
orthoquinones, which polymerize non-enzymatically to
melanin – Melanization.
• These reactions are accompanied by local production
of cytotoxic free radicals, quinones or semi quinones
that contribute to killing microorganisms or parasites.
 Cellular Immunity
• Lamellocytes are rarely seen in healthy insects.
• Its precursors are restricted to the lymph glands and
differentiate upon parasitization by invaders too large
to be engulfed by plasmatocytes.
• Lamellocytes are large, adhesive flat cells that associate
with one another to form capsules around invaders.
• Upon infection a parasite is first recognized by
plasmatocytes which bind to the invader and sends
differentiation signal to the lymph gland.
• The lamellocytes are produced and released from the
lymph glands, bind to the invader and form a
multilayered capsule around it.
 Cellular Immunity
• Signals created by the plasmatocytes also increase
number of crystal cells – secrete PO which catalyze
melanization of the resulting capsule.
• The parasite is killed by asphyxia or by local production
of toxic compounds in the course of melanization.
• Hemocytes also participate in the control of infection
by production of secreted molecules including
thioester-containing proteins (TEPs) which act as
opsonins to promote phagocytosis.
• Opsonin is an antibody or any other molecule that bind
to foreign microbes or cells making them susceptible to
phagocytosis.
 Response to Viral Infection
• Honey bees like other arthropods present several mechanisms
to control infections by viruses.
• An evolutionarily conserved mechanism they use to counter
viral infection is programmed cell death (Apoptosis).
• Apoptotic response affect viral replication and gene expression
and hence control the infection.
• However, some viruses (i.e. baculovirus) encode apoptotic
suppressor gene p35 that prevents apoptosis and allows the
virus to replicate and spread.
• Blood cells play a role to control baculoviruses by
encapsulating the infected cells.
• In some cases, secreted Phenoloxydase has also shown
antiviral activity against baculoviruses.
 Response to Viral Infection
• Another host-defense mechanism against viruses is RNA
interference.
• Involves RNA degradation triggered by double stranded
RNA

(RNA induced silencing complex)

 JAK/STAT Signalling Pathway
• This pathway targets activation of a transcription factor
STAT92E which induce expression of a number of genes
responsible for defense against viral infection
 Functions of JAK/STAT regulated humoral
factors
• TEP1 function as Opsonin that promote
phagocytosis as well as protease inhibitor

• totA enhance response to stress and confers

longer survival in stress conditions

• Raf induces hemocytes proliferation and hence

enhances cellular responses particularly
encapsulation and phagocytosis