Research Paper

Mediators of Inflammation 2, 217-223 (1993)

THE time course of effect of the B2-receptor agonist kalli-din (K) on induced changes of nasal airflow, rhinorrhoea,nasal pain, sneezing and nasal microvascular leakage hasbeen examined and compared with its B2 metabolite agon-ist bradykinin (B) and the Bl-agonist [des-argg]-bradykinin (D). When administered as a single dose K and Binduced an immediate sensation of pain, rhinorrhoea,elevations in lavage albumin and protein levels and asustained increase in nasal airways resistance (NAR) for5-40 min post-challenge. [des-argg]-Bradykinin and ve-hicle placebo (V) were without effect on any of theseindices. These studies identify the action ofK and B withinthe nose and differentiate the neural and vascular effectsof these kinins in addition to suggesting the potential thatnasal blockage and nasal microvascular leakage representalterations in differing vascular compartments. These find-ings have implications for the understanding and thera-peutic manipulation of rhinitis.

Key words: Bradykinin, [des-arg9]-Bradykinin, Kallidin

The temporal relationshipbetween the neural andvascular actions of kallidinwithin the nose

K. Rajakulasingam,cA L. C. K. Lau,R. Polosa, M. K. Church, S. T. Holgate andP. H. Howarth

Immunopharmacology Group, Centre Block,Southampton General Hospital, Tremona Road,Southampton SO9 4XY, UK

CA Corresponding Author

Introduction

The introduction of potent and specific HI-antihistamines in the 1940s, confirmed histamine asan important mediator of allergic rhinitis but, inproducing only partial relief of symptoms, revealedthe participation of other mediators in this disease.Among these are the kinins, including, bradykininand kallidin (lysyl bradykinin), both of which arepotent vasoactive peptides formed as cleavageproducts from the action of kallikreins on high andlow molecular weight kininogens respectively.Metabolism of bradykinin occurs with cleavage ofthe N-terminal [des-arg9] residue to a peptide whichis reported to be active on the B-receptor subtype.2

The presence of increased kinin recovery by nasallavage during the early3 and late4 phases ofallergen-provoked rhinitis and also in naturallyoccurring rhinitis provides evidence that kiningenerating systems are operative in this disease andtheir involvement in symptom generation issuggested from nasal challenge studies. Nasalinsuflqation with bradykinin elicits many of theclinical features of rhinitis including nasal blockage,rhinorrhoea and increased micro-vascular leak-age.6’7 Less is known, however, of the nasal effectsof kallidin.

Kallidin comprises approximately half of theamount of kinin recovered following nasal allergenchallenge3 and accords with the finding of increasedtissue kallikrein in lavage fluid. Concordant withthis, elevated levels of aminopeptidase, capable ofconverting kallidin to bradykinin, have beendemonstrated in post-allergen challenge nasallavage.9 Bradykinin and kallidin both produce nasal

(C) 1993 Rapid Communications of Oxford ktd

blockage by interacting with the B2-receptorsubtype and are of comparable potencies whengiven in incremental doses.7 However, in the lowerairways, kallidin is about three times lss potent asa bronchoconstrictor than bradykinin in molarterms. 10

To further explore the comparative range ofeffects of kallidin within the nose, the time courseof action of this kinin on individual nasal symptomsand signs has been studied in parallel withcomparable amounts of the nasal effects of the B2

agonist bradykinin and the B agonist [des-argg]bradykinin in healthy volunteers.

Subjects and Methods

The study was conducted in eight non-rhinitic,non-atopic healthy volunteers (six males and two

females) with a mean age of 29 years. All subjectsrefrained from taking any medications throughoutthe study period. None of the subjects had a historyof recent upper respiratory tract infection, nasalpolyps, infective rhinitis, nasal surgery or nasaldeformities and all gave written informed consent.The study was approved by the Southamptonhospitals and University joint ethical subcommittee.The study was conducted in three phases.

Phase I: Subjects attended on four occasions at thesame time of the day to receive nasal challenge withincremental doses of either bradykinin, kallidin,[des-argg]-bradykinin or vehicle placebo in adoubl’e-blind cross-over manner. The objectiveresponse parameter was nasal airways resistance(NAR). At each visit, subjects were rested for 15

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K. Rajakulasingam et al.

min and baseline measurements of NAR were madeby active posterior rhinomanometry using a

Mercury NR6 Rhinomanometer (Mercury instru-ments, Glasgow, Scotland) as described previously.7

The measurement of NAR has a coefficient ofvariation of repeated measurements of 10.7%.Providing the baseline values of NAR did not varyby > 10%, subjects then underwent nasal challengewith vehicle followed by incremental doses of eitherbradykinin, kallidin, [des-arg9]-bradykinin orvehicle placebo at 20 min intervals until the topchallenge dose had been achieved. The three kinins

(Nova Biochem Ltd, Nottingham, UK) weredissolved in ethanol (10%) and 0.9% sodiumchloride to produce concentrations of 0.077, 0.77,3.85 and 7.7 mg/ml. These doses were chosen fromprevious studies. 6’7 The purity of the synthetickinins was confirmed by high-performance liquidchromatography (HPLC) as described previously.Nasal challenge was undertaken bilaterally using ahand-held pump spray delivering 0.13ml peractivation with a coefficient of variation of outputof 8.4%. The spray was placed in one nostril whileoccluding the other and activated once during quietinspiration. The procedure was then repeated in theopposite nostril. At the concentrations employed,this method delivered total kinin doses of 20, 200,1 000 and 2 000 g equally divided between the twonostrils. After each challenge repeat measurementsof NAR were made at 3, 5, 10, 15 and 20 minpost-challenge.

Phase 2: To investigate the time course of effect ofnasal insufflation with kinins on induced symptomsand signs, each of the eight subjects attended at thesame time of the day on four further occasionsseparated by at least 7 days to receive nasal chal-lenge with a single dose (2 000 #g) of bradykinin,kallidin, [des-arg9]-bradykinin or vehicle in adouble-blind crossover manner. Repeat measure-ments of NAR were made at 3, 5, 10, 15, 20, 25,30, 35 and 40 min post-challenge. Subjects alsorecorded their symptom of nasal pain/discomfort ona 10 cm visual analogue scale, before and at 3, 5,10, 15, 20, 25, 30, 35 and 40 min post-challenge.Rhinorrhoea was measured 45 s before each of thesetime points by asking the subjects to blowsecretions into a pre-weighed tissue towel. Thetissue towels were then immediately reweighed andthe weight of secretion calculated by subtraction.The symptom of sore throat was recorded aspresent or absent and the number of sneezesfollowing challenge was counted.

Phase 3: To investigate the time-course of action ofthese kinins on nasal vascular permeability, each ofthe eight subjects attended on an additional fouroccasions, and were randomly allocated to receivenasal challenge with the same single dose of

bradykinin, kallidin, [des-arg]-bradykinin orvehicle as for phase 2, in a double-blind, cross-overstudy design. At each attendance sequential nasallavage was performed bilaterally five times, at 1-minintervals, prior to nasal insufflation challenge.Subjects were instructed to tilt their headsbackwards about 30 from the horizontal while inthe sitting position, to hold their breath and refrainfrom swallowing. Two and a half ml of 0.9%prewarmed saline was instilled into each nostril andafter 10 s subjects flexed their necks and expelledthe mixture of airways surface liquid and saline intoa collection vessel. Nasal lavage was then repeatedat 3, 5, 10, 15, 20, 25, 30, 35 and 40 min after eachof the nasal challenges,

Analysis of nasal lavage )quid: Lavage samples wereimmediately stored on ice until the conclusion ofthe experiments when they were frozen at --20Cto await protein analysis. After thawing at roomtemperature, total protein concentrations weremeasured in 0.25 ml replicates by the method givenby Bradford11 using Coomassie Blue G-250 reagent(Pierce, Rockford, IL, USA) as indicator. Absor-bance read at 595 nm was compared with a standardcurve constructed with bovine serum albumin(Sigma, Poole, UK) to give absolute proteinconcentrations.Albumin concentrations were measured by

rocket immunoelectrophoresis. 12 Three microlitrealiquots of nasal lavage fluid were introduced intowells cut in agarose gel and subjected to

electrophoresis (4 V/cm) for 20 h into a ’window’of agarose containing 1.18ktl/cm2 of rabbitanti-human albumin antibody (Nordic, Tilburg,Netherlands). Immunoprecipitates were stainedwith Coomassie Brilliant Blue. Albumin levels werequantified by comparison with standard curvesconstructed using human serum albumin (Sigma,Poole, UK). The sensitivity of the albumin assaywas 1 #g/ml and the coefficient of variation ofrepeated measurements is 5%.

Data anasis: On the different challenge days baselinemeasurements of NAR, lavage albumin and totalprotein were compared using the Friedman’stwo-way ANOVA test. The time-dependentchanges in NAR (Pa/cm3s), lavage albumin andtotal protein were compared both within groupsand between groups using Friedman’s test. Toassess differences between treatments, the mean areaunder the curve (AUC) for each time period ofobservation was calculated and compared using theWilcoxon’s signed rank test for paired data. TheWilcoxon’s signed rank test was also used forcomparisons of the challenges on the quantity ofnasal secretions, the number of sneezes and thevisual analogue scores for nasal pain. Values are

218 Mediators of Inflammation. Vol 2.1993

Nasal responses to kinins

expressed as mean-F standard error of mean ormedian values with ranges.

Results

Phase 1: There were no significant differencesbetween the baseline NAR measurements in theeight subjects on any of the four challenge days(p > 0.05). Both bradykinin and kallidin caused a

dose-dependent increase in NAR (p < 0.01),while [des-argg]-bradykinin and vehicle werewithout significant effect (Fig. 1). There was no

significant difference in the eects of bradykininand kallidin on NAR at any dose level > 0.05)whether assessed as peak response or area underthe curve.

Phase 2: There were no signifi’cant differencesbetween baseline N.AR measurements in the eightsubjects on any of the four challenge days(p > 0.05). Insuf]:lation of vehicle had no signifi-cant effect on NAR. Bradykinin administered as a

single dose of 2000/g caused a rapid increase inNAR reaching maximum of 71.4% above baselineat 15 min (p < 0.05) and gradually declining over40 min to 50.0% above baseline (Fig. 2). Kallidinalso increased NAR by a median maximum of57.1% (p < 0.05). This peak was reached 20 minpost-challenge. The increase in NAR with challengewas significantly greater for bradykinin thankallidin at 5 and 15 min post-challenge butthereafter there was no significant difference fromthat seen with bradykinin (Fig. 2). However, therewas no significant difference between the areasunder the time-response curves for both agonists(p > 0.05). The increase in NAR was significantlygreater than the pre-challenge measurements from3 to 40 min (p < 0.05). In contrast [deg-argg]-bradykinin produced no significant change in NAR.

0.6

0,5

mg 2 mlgDil 20 /Jg 200 ,g \ [I %

0 10 20 30 40 50 60 70 80 90 100 110 120

Time (min)

FIG. 1. Dose-dependent changes in NAR following nasal challenge withincremental doses (20, 200, 000 and 2 000 #g) of bradykinin (O),kallidin (/k), [des-argg]-bradykinin (V) and 10% ethanol saline (O).Each result is the median of observations in eight subjects.

0.30

E 0.25

0.20

0.15

0.10

0 10 15 20 25 30 35 40

Time Imin)

FIG. 2. NAR changes following nasal challenge with single dose ofbradykinin 2 000 #g (O), kallidin 2 000 #g (/k), rdes-arg9]-bradykinin2 000 #g (V) and 10% ethanol saline (O). Each result is the median ofobservations in eight subjects.

When compared with vehicle, bradykinin andkallidin, but not [des-argg]-bradykinin, producedrhinorrhoea, with median total secretion weights of376 (range 179-597) mg and 330 (range 60-1351) mg, respectively (p < 0.05). The increase innasal secretions was maximum 3 min post-challengeand declined rapidly to very low levels,indistinguishable from placebo, by 20 min post-challenge (Fig. 3A). Both bradykinin and kallidinproduced similar degrees of nasal pain, with median(range) values of 3.45 (range 0.6-7.2)cm and 3.4(range 0-7.5)cm, respectively (p < 0.05). Nasalpain reached maximum at 3 min post-challenge andlasted for 5 rain (Fig. 3B). Neither [des-argg]bradykinin nor vehicle produced algesic effect (Fig.3B). None of the subjects complained of nasal itchand following bradykinin and kallidin challengethree subjects complained of sore throat lasting upto 25 min. None of the subjects sneezed followingkinin nasal challenge.

Phase 3" There were no significant differencebetween the fifth baseline nasal lavage albuminlevels in the eight subjects on the four challengedays, the group means being 24.73 5.71,26.14-F 10.1, 27.71 -F 12.9 and 27.7-F 9.14 #g/mlfor subjects receiving bradykinin, kallidin, [des-arg9]-bradykinin and vehicle, respectively. InsuflClation with bradykinin and kallidin increased theconcentration of albumin recovered in the nasal

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K. Rajakulasingam et al.

160 3.5

E

140 ) 3.0

O

20 .::: 2.5

oo2.0

80

1.5

(B)

Time (min) Time (min)FIG. 3. Effects of bradykinin 2 000 #g (O), kallidin 2 000 #g (/k), [des-argS]-bradykinin 2 000 #g (V) and 10% ethanol saline vehicle((C)) on (A) rhinorrhoea, (B) nasal pain. Each result is the median of observations in eight subjects.

lavage (p < 0.05) (Fig. 4A). On repeated lavage,maximum albumin concentration achieved withboth challenges occurred within 10 min anddecayed progressively to baseline by 40 min (Fig.4A). By contrast nasal challenge with [des-arg9]

bradykinin and vehicle failed to induce significantchanges in lavage albumin concentration (Fig. 4A).

There were no significant differences between thefifth baseline nasal lavage total protein levels in theeight subjects on the four challenge days, the group

sso (A)

2OO

150

100

SO

0

I’5 "0 10 20 25 30 35 40

400

350

300

250

o 200

0

" s0

0

100

50-

(B)

00 10 15 20 25 30 35 40

Time (min) Time (min)

FIG. 4. Nasal lavage (A) albumin and (B) total protein changes following nasal challenge with single dose of bradykinin 2 000 #g (O), kallidin 2 000/g(/k), [des-argS]-bradykinin 2 000 #g (V) and 10% ethanol saline ((C)). Each result is the mean +__ S.E. of mean of observations in eight subjects.

220 Mediators of Inflammation. Vol 2.1993

Nasal responses to kinins

means being 121.58 _--+- 40.14, 101.54 23.12,67.25 __+ 8.63 and 90.25 -+- 20.88 #g/m1 for sub-jects receiving bradykinin, kallidin, [des-arg9]bradykinin and vehicle, respectively. When com-pared to vehicle, both bradykinin and kallidininduced significant increases in lavage total proteinlevels (p < 0.05) with a time course similar to thatseen with albumin whereas [deg-argg]-bradykininhad no effect (Fig. 4B). The albumin and totalprotein responses following bradykinin and kallidinwere not significantly different when comparedusing the AUC (p > 0.05).

Discussion

This study extends and confirms our previousfindings that Be-agonists bradykinin and kallidininduce a dose-dependent increase in nasal airwaysresistance while the Bl-agonist [des-arg9]-bradykinin is without effect. 7 By selecting equi-doses ofkallidin and bradykinin it has been possible to

further explore their time course of action and to

investigate their relative effects not only on nasalblockage but also on nasal pain, sneezing,rhinorrhoea and on increments in nasal lavagealbumin and total protein levels. Both kallidin andbradykinin but not [des-arg9]-bradykinin inducedchanges in all these indices with the exception ofsneezing.Within human lower airways kallidin is three

times less potent than bradykinin as a bronchocon-strictor when these kinins are compared on a molarbasis.1 In contrast we have previously found thatkallidin and bradykinin, when administered inincremental doses, to be approximately equipotentin the nasal obstructive effect within the upperairways.7 Similar differences have been reported inother species and our finding is not inconsistentwith other studies where bradykinin and kallidinhave been shown to have varying potency ofaction. 3’4 In rats, kallidin has been shown to bemore potent than bradykinin in causing relaxationof isolated smooth muscle preparation fromduodenum, although on isolated smooth musclesfrom guinea-pig ileum and rat uterus a reversedorder of potency has been observed.3 Kallidin isalso more potent than bradykinin as a localvasodilator in dogs.4 In the canine trachealepithelium, which is enriched in kinin receptors,kallidin is equipotent with bradykinin in inducingchloride secretion, is The relevance of such aconsideration to rhinitis is the identification thatkallidin represents 45% of the total kinin pool inlavage fluid following nasal allergen challenge.Single dose nasal administration of both kallidinand bradykinin but not [des-argg]-brady-kinininduced a significant increase in nasal airways

resistance. For each of the B2-receptor agonists thiswas significant from 5 to 40 min post-challenge(Fig. 2). Although the effect of bradykinin appearsgreater than that of kallidin there was no significantdifference between the areas under the time-response curves for both agonists. However, whencompared at individual time points, the NARincrease following bradykinin nasal challenge was

significantly greater at 5 and 15 min post-challenge.The peak NAR responses following kallidin andbradykinin were at 20 and 15 min post-challenge,respectively.The present study also identifies that both

kallidin and bradykinin induce nasal pain, rhi-norrhoea, nasal blockage and an increase in nasallavage protein levels. These are the first humanstudies investigating in detail these neural andvascular mediated effects of kallidin within the nose.The nasal blockage and increase in lavage proteinlevels are considered to be manifestations of directvascular actions. Inconsistent with this, kininB2-receptors have been localized to the vasculatureof the nasal mucosa,6 and the nasal obstructiveresponse, but not the plasma protein exudation, hasbeen shown to be prevented by prior administrationof the -adrenergic agonist oxymetazoline. It istherefore likely that nasal obstruction and plasmaprotein extravasation represent different vascularmechanisms. Consistent with this, the time courseof increase in lavage protein levels which peaks at

10 min and resolves within 20 min post-challengediffers from the time course of effect on nasalairflow, further suggesting that the nasal blockageand plasma protein leakage are two independentvascular effects (Figs 2 and 4). Nasal obstructionarises from engorgement of venous plexi within theturbinates.8 These plexi are under sympatheticregulation and factors which increase sympatheticstimulation (i.e., exercise, 0-adrenergic agonists)increase nasal airflow and reduce nasal airwaysresistance whereas mediators, such as histamine,prostaglandins and kinins which have directvasodilator properties induce nasal obstruction.

In addition, laser doppler flowmetry studies haveidentified changes in blood flow in the superficialmucosal vasculature following bradykinin challenge(Rajakulasingam K, unpublished findings). This isnot always in concordance with the nasalobstructive response and it is likely that endothelialcontraction with opening of gap junctions inpost-capillary venules present beneath the basementmembrane and around glands in the nasal mucosa,are responsible for the increase in lavage proteinlevels.9 Consistent with this, changes in vascularpermeability produce unrestricted plasma molecularleak, concordant with the albumin and total proteinincrements in lavage with kallidin and bradykinin.Although it is not possible to exclude glandular

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K. Rajakulasingam et al.

secretion of protein, this is less likely as our studyshows that the predominant protein in nasal lavageis albumin (Fig. 4), a plasma protein synthesized byliver and no Be-receptors have been localized to

glandular structures within the human nose. 16 Theneural effects of kinin insufflation are pain andpossibly rhinorrhoea. With histamine nasal insutTla-tion the rhinorrhoea is bilateral following unilateralchallenge and can be inhibited by antimuscarinicpretreatment indicative of a reflex vagal mechan-ism.2

In a separate investigation we have been unableto identify any protective effect of nasal ipratropiumbromide on bradykinin induced rhinorrhoea.21 Thissuggests that the mechanism of rhinorrhoea withbradykinin differs from histamine. Consistent withthis the sensory neural stimulation induced bykallidin and bradykinin differs from histamine is notinducing sneezing. Thus the nasal pain induced bykinins must reflect different sensory neuralpathways than those stimulated by histamine. Thatthese nerves are distinct from the classicalunmyelinated ’C’ sensory nerves is furthersuggested by the identification that the ipsilateralsecretory effects of unilateral capsaicin nasalchallenge are blocked by the coadministration ofintramuscular atropine and topical ipratropiumbromide22 and that repeated nasal capsaicinpretreatment has failed to diminish the algesicresponse to kallidin.23 This indicates that the humannose differs from animals in its response to kinins,as the effects of bradykinin in canine and rat airwayshave been shown to be mediated via capsaicinsensitive, sensory neurones,24’25 and further high-lights the interspecies differences in response to

kinins.It is possible that the rhinorrhoea induced by

kallidin and bradykinin which peaks at the sametime as the nasal pain following challenge (Fig. 3)represents stimulation of neuropeptide containingnerves with consequent peptide release andglandular secretion. Alternatively, as bradykinin hasbeen reported to stimulate prostaglandin synth-esis26’27 it is possible that this could account for thenociceptive and secretory effects of kallidin andbradykinin. Against such a possibility is the rapidityof onset of the algesic effect, and the recent reportthat oral acetyl salicylic acid had no effect on thenasal discomfort following nasal bradykinin chal-lenge. 17 A further possibility is that the rhinorrhoeais a reflection of plasma leakage rather thanglandular secretion. While studies in cats and ferretshave indicated that apparent secretion induced bybradykinin is not associated with an increase inmucous glycoconjucates16 the rapid increase inrhinorrhoea observed in this study, which occurredbefore marked increases in lavage albumin wereobserved, would argue against such a possibility.

Thus the exact mechanism of the rhinorrhoearemains to be elucidated.

In conclusion this study draws attention to thevascular actions of kallidin and bradykinin withinthe nose and indicates that obstruction and vascularpermeability represent distinct entities. The pro-longed action of these kinins on nasal obstructionsuggests their importance in clinical disease.However, the contribution of kinins to thesymptomatology of rhinitis will not be fullyelucidated until potent and selective inhibitors ofkinin generation or antagonists of kinin receptormediated effects become available.

References1. Proud D. Kinin formation: mechanisms and role in inflammatory disorders.Annu Rev Immunol 1988; 6: 49-83.

2. Regoli D, Barabe J. Pharmacology of bradykinin and related kinins.Pharmacol Rev 1980; 32: 1-46.

3. Proud D, Togias A, Naclerio RM, Crush SA, Norman PS, Lichtenstein LM.Kinins generated in vivo following nasal airway challenge of allergicindividuals with allergen. J Clin Invest 1983; 72: 1678-1685.

4. Naclerio RM, Proud D, Togias AG, et al. Inflammatory mediators in lateantigen-induced rhinitis. N Engl J Med 1985; 313: 65-70.

5. Svennsson C, Andersson M, Persson CGA, Venge P, Alkner U, Pipkorn U.Albumin, bradykinin and eosinophil cationic protein the nasal mucosalsurface in patients with hay fever during natural allergen exposure. J AllergyC/in Immunol 1990; 85: 828-833.

6. Proud D, Reynolds CJ, Lacapra S, Sobotka AK, Lichtenstein LM, NaclerioRM. Nasal provocation with bradykinin induces symptoms of rhinitis and

throat. Am Rev Respir Dis 1988; 137: 613-616.7. Rajakulasingam K, Polosa R, Holgate ST, Howarth PH. Comparative nasal

effects of bradykinin, kallidin and [des-argg]-bradykinin in atopic rhinitis andnormal volunteers. J Physiol 1991 437: 577-587.

8. Baumgarten CR, Nichols RC, Naclerio RM, Proud D. Concentrations ofglandular kallikrein in human nasal secretions increase during experimentally-induced allergic rhinitis. J Immuno11986; 137: 1323-1328.

9. Proud D, Baumgarten CR, Naclerio RM, Ward PE. Kinin metabolism innasal secretions during experimentally-induced allergic rhinitis. J Immunol1987; 138: 428-434.

10. Polosa R, Holgate ST. Comparative airway response to inhaled bradykinin,kallidin and [des-argg]-bradykinin in normal and asthmatic subjects. A RevRespir Dis 1990; 142: 1367-1371.

11. Bradford MM. A rapid and sensitive method for the quantitation ofmicrogram quantities of protein utilizing the principle of protein-dye bind-ing. Anal Biochem 1976; 72: 248.

12. Weeke B. Rocket immunoelectrophoresis. Scand J Immuno11973; 1: 37-46.13. Reis ML, Okino L, Silva MR. Comparative pharmacological actions of

bradykinin and related kinins of larger molecular weights. Biochem Pharmacol1971 20: 2935-2946.

14. McCarthy DA, Potter DE, Nicolaides ED. An in vivo estimation of thepotencies and half-lives of synthetic bradykinin and kallidin. J Pharmacol ExpTherap 1965; 148: 117-122.

15. Rangachari PK, McWade D, Donoff B. Luminal receptors for bradykininthe canine tracheal epithelium: functional subtyping. Regul Peptides 1988;

21 237-244.16. Baraniuk J, Lundgren JD, Mizoguchi H, et al. Bradykinin and respiratory

membranes. Am Rev Respir Dis 1990; 141: 706-714.17. Churchill L, Pongracie JA, Reynolds CJ, Naclerio RM, Proud D. Pharmaco-

logy of nasal provocation with bradykinin: studies of tachyphylaxis, cycloox-ygenase inhibition, -adrenergic stimulation and receptor subtype. Int ArchAllergy Appl lmmunol 1991 95(4): 322-331.

18. Eccles R. Neurological and pharmacological considerations. In: Proctor DRand IB Anderden, eds. The Nose: Upper Airway Physiology and the A tmosphericEnvironment. Amsterdam: Elsevier, 1982; 191-214.

19. Cauna N, Hinderer KH. Fine structure of blood vessels of the human nasalrespiratory A Otol Rhinol Laryngol 1969; 78: 865-879.

20. Mygind N, Secher C, Kirkegaard J. Role of histamine and antihistamines inthe Eur Respir J 1983; 64 (suppl 128): 16-20.

21. Rajakulasingam K, Polosa R, Lau LCK, Church MK, Holgate ST, HowarthPH. The influence of terfenadine and ipratropium bromide alone and incombination bradykinin induced nasal symptoms and plasma proteinleakage. Clin Exp Allergy 1992; 22: 71%723.

22. Stjarne P, Lundblad L, Lundberg JM, Anggard A. Capsaicin and nicotine-sensitive afferent and nasal secretion in healthy human volunteersand in patients with vasomotor rhinitis. Br J Pharmaco11989; 96: 693-701.

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23. Geppetti P, Fusco BF, Alessandri M, et al. Kallidin applied to the humannasal produces algesic response not blocked by capsaicin densensiti-zation. Regulatory Peptides 1991; 33: 321-329.

24. Kaufman MP, Coleridge HM, Coloeridge JCG, Baker DG. Bradykininstimulates afferent vagal C-fibres in intrapulmonary airways in dogs. J ApplPhysiol 1980; 48: 511-517.

25. Lundberg JM, Saria A. Capsaicin induced desensitization of the airwayto cigarette smoke, mechanical and chemical irritants. Nature (Lond)

1983; 302: 251-253.26. Leikauf GD, Ueki IF, Nadel JA, Widdicombe JH. Bradykinin stimulates

chloride secretion and prostaglandin E2 release by canine tracheal epithelium.Am J Physiol 1985; 248: F48-55.

27. Conklin BR, Burch RM, Steranka LR, Axelrod J. Distinct bradykininreceptors mediate stimulation of prostaglandin synthesis by endothelial cellsand fibroblasts. J Pharmacol Exp Therap 1988; 244: 646-649.

ACKNOWLEDGEMENTS. We grateful to Dr Andrew Walls for histechnical advice protein assay and Dr Doreen Ashworth (Ferring Institute)for HPLC analysis of the purity of kinins. This study supported by grantfrom the Ferring Peptide Research Partnership, Malmo, Sweden.

Received 23 February 1993"accepted in revised form 16 March 1993

Mediators of Inflammation. Vol 2.1993 223