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The Th1 subset and increased IFN are important for the control of Leishmania ,
T. cruzi and
Toxoplasma infections, whereas the Th2 response is more important in parasitic
infections in which antibody is a major factor.
Immune pathology
The protozoa can elicit humoral responses in which antigen antibody complexes
are formed and these can trigger coagulation and complement systems.
Immune complexes have been found circulating in serum and deposited in the
kidneys where they may contribute to conditions such as glomerulonephritis.
Another important form of antibody mediated pathology is autoimmunity
These autoantibodies may play a role in the pathology of parasitic diseases by
exerting a direct cytotoxic effect on the host cells
A range of parasite - derived enzymes such as proteases and phospholipases can
cause cell destruction, inflammatory responses
Immune evasion
Mechanisms that allow evade the host immune response
- Antigenic masking and Antigenic variation
Masking means the parasite becomes coated with host components and therefore
is not recognized as foreign
In addition parasites can undergo antigenic variation which results in surface
antigens being changed during the course of an infection
Detection of parasites
The most commonly used method involves microscopy and this can be applied to
clinical as well as environmental samples.
For some organisms this approach remains the gold standard for detection and
identification.
The advantages of microscopy are speed, cost and availability.
In addition, fluorescent - labelled antibodies raised to species - specific antigens can
be utilized to help identify organisms to species or subspecies level, and other
stains can be used to help determine the viability of cells.
Fluorescein diacetate which is cleaved by esterases in viable cells, releasing
fluorescein (gives a green fluorescence) and propidium iodide which is excluded
from viable cells but taken up by cells with damaged membranes (gives red
fluorescence)
Antibody - based technologies
The most commonly used method is the ELISA and this can be used to detect the
presence of antigens in samples (direct assay) or antigen - specific antibodies in
patients serum (indirect)
Has good specificity and sensitivity
DNA - based technologies
The advent of highly sensitive DNA amplification technologies such as the
polymerase chain reaction (PCR) has allowed the development of molecular tools for
the identification and detection of parasites
Other DNA markers that have been used included genes that encode for metabolic
enzymes and
structural proteins.
For example Cryptosporidium can be speciated by using a combination of target
genes including the Cryptosporidium oocyst wall protein (COWP), heat shock
protein (hsp70), dihydrofolate reductase (DHFR) and 18S ribosomal DNA
Other DNA - based techniques are now available such as the multiplex PCR
(amplification of more than one gene at a time) and quantitative real - time PCR
(qPCR), and these have improved specificity and sensitivity in detection and
increased the usefulness of DNA technology for the detection of parasites
A recent method, DNA microarray, is a fast developing technology. This is a chip
based system that measures the binding of target DNA sequences (parasite) from
samples (fluorescently labelled using PCR) to complementary sequences bound to
the chip surface. The binding of parasite sequence DNA and the amount that is
bound can be detected using fluorescence
Alternative methods
Clinical samples
The type of samples can vary from blood where preparation can be minimal (e.g.
sample of smears for microscopy) to faeces or intestinal aspirates
Sedimentation method using the formol ether or formol ethyl acetate
techniques
Flotation method using copper sulphate
Environmental samples
This section focuses on water and foods as they are often major routes of
transmission for parasites such as Giardia and Cryptosporidium which are included
in the monitoring standards for potable waters
Food and pharmaceutical companies that utilize water in their processes are
required to test for a number of pathogens
If levels of a parasite in water are low (< 1/100 ml) several litres must be collected
and analysed.
This is possible, but would require the sample volume to be reduced for analysis.
This can be achieved either by filtration or centrifugation.
If the sample contains sediment, both of these approaches are difficult; however,
methods such as tangential flow filtration (fluid passed parallel to the filter) can be
used
Chemotherapy
There has been slow progress in the development of new and novel antiprotozoal
agents
Interestingly there are still a number of protozoan parasite infections such as
cryptosporidiosis for which there is no effective treatment
Mechanisms of action and selective toxicity
Dapsone and the sulphonamides - block the biosynthesis of tetrahydrofolate by
inhibiting dihydropteroate synthetase