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methodology of sputum induction and processing" @jukanovic, Sterk et al. 2002) which has
a detailed report on sputum induction @aggiaro, Chanez et al.2002), methods of processing
@fthimiadis, Spanevello et al. 20f.2) and measurement of fluid-phase mediators (Kelly,
Keatings et aI,2002) concentrating on techniques and findings in asthma and COPD subjects.
Pre-treatment with p2 agonists as appropriate and lung function and/or oxygen saturation
monitoring is recommended throughout the procedure @aggiaro, Chanez et al.2002).
The eady studies confirmed and enlarged on the inflammatory profiles seen in asthmatics
compared to normal subjects as had been described in the biopsy and lavage studies above.
An eosinophilic dominant inflammation was seen with increases in total cell numbers, as well
as individual eosinophil and neutrophil counts, eosinophil derived proteins [ECP, eosinophil
derived neurotoxin (EDNI and major basic protein (MBP)1, (Pin, Gibson et al. 1992; Fahy,
Liu et al. 1993; Foresi, I-eone et al. 1997; Louis, Shute et al. 1997; Bacci, Cianchetti et al.
1998; Park, Whang et al. 1998; Rosi, Ronchi et al. 2000) the allergic profile interleukins (IL3,
nA, L5, Ll0) and certain proteins such as tryptase, eosinophilic chemo-attractant factors,
and adhesion molecules (Shoji, Kanazawa et al. 1998; Hamzaoui, Brahim et al. 2000).
Correlations between these sputum inflammatory cells and markers and clinical parameters
have been investigated. The strongest associations appear to be during an acute asthma
exacerbation with increases in eosinophils, activated eosinophils and ECP (Jang and Choi
2000; Jang, Choi et al. 2000), basophils and mast cells @in, Freitag et al. 1992; Gauvreau,
I*e et al. 2000). During times of increased symptoms, eosinophil number, ECP, IL5 and
albumin were all found to independently contribute to abnormalities of FEVr and FVC in
asthmatics (Fujimoto, Kubo et al. 1997; Bacci, Cianchetti et al. 1998; Shoji, Kanazawa et al.
1998; Grebski, Wu et al. 1999; Woodruff, Khashayar et al. 2001; Bartoli, Bacci et al.2N4).
Severe asthma was predicted by high eosinophil numbers, ECP and albumin concentrations,
though there was no such gradation in mild to moderate asthmatics (Fujimoto, Kubo et al.
1997; Bartoli, Bacci et aL.2ffi4). The eosinophil counts gave a sensitivity of 68.3Vo, and a
specificity of 55.3Vo for moderate to severe asthma compared to mild, and was of a higher
diagnostic value for asthma than sputum ECP or methacholine challenge (Park, Whang et al.
1998; Rosi, Ronchi et al. 2000).
I 'Eosinophil protein X' and 'eosinophil derived neurotoxin' is the same protein but referred to by these two
different names throughout studies, and denoted as eitler 'EPX' or 'EDN'. I have elected to refer to it as
'eosinophil derived neurotoxin' and 'EDN'. Eosinophil cationic protein, eosinophil derived neurotoxin, major
basic protein and eosinophil peroxidase are all separate proteins.
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