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65 Prolonged release of histamime from human basophils
VOLUME NUMBER
Abstracts
87 1, PART 2
67
65
155
E CANINE BRONCU
Phil&lphi8Jk Todetermineif antigen&E intemctionr cm induce prolonged bosophil histaminereleaae@Wve studiedthe tem00raloatterneof %BfiRin 10a&mice8fter serial and&n,d-10 pm/ml) incubations &rr 4 hours (Table1) TIHEWURS) ;L 1 2 llt2 7tl 6al 39iaf 11*212*2= i.1 6Oi7' 1 16t3* a*2 l&4* 8t2 ~0p~0.01vsOPSN2Uf~p~005vsOPNU
fllmsi
9 9t2 7il lOi 7i2
Whilethe maimurn BkRoccukedin the 1sthr significant increasesof BERoccurredafter the 1sthr at eachof the antigen doses.Wealsocompued the sum of the BHRafter antigen in : ahhe 1sthr $1 m undisturbed fashion from 2-5hrs md c) serial removalandrexposure to fresh antigen(2,3,4hrs), Table2. TABLE2. TM (HOURS) 1 lOt2 39t9' 59t7' :0 5lt6* l p~O.OlvsOPNU,,=P
F 0.1
lit1 = z 35t6*r' 19t5 19t5 3%t? 19*3* < O.Olvs2-5HR5 Serialreplacement of Wigen al 2.3 and 4 hrs resultedin 37-85X highercumuluive BHRuUnthe2-)hr
undklurbed incubations.Furthermomprolonged ewosun to antigen by either methodresultedin an additional3230%BRRalter the 1 hr incubations. Theseobsemationsmayexplain the pattern of prolonged HRve have previously observed during prolonged antigen exposurein cutaneousallergic reactions.
66
-Cl.
OF v KERATINGCVED RP&AXING FACTOR. w M.D.. D. N, Wtieht. M.D.. D. K. Ledford. M.D.. J. J. Krzanowski, Ph.D.. H. A. Sh&ay&* M.D.. Ph.D.. R. F. Lw Tampa, Florida and ‘Zagazig. Egypt Previous work has shown that the in vitro removal of epithelium from canine bronchial airways increases the hyperresponsiveness of the bronchial smooth muscle (BSM) to acetylcholine (ACh) due to the loss of epithelium-derived relaxing factor. We have previously demonstrated that cultured murine keratinocyte suspension supernatant (MKSS) produces a non-prostanoid factor that reduces the AChinduced contraction of de-epithelialized (DE) canine BSM. The effect of 60° and 90°C on this factor was determined. Murine keratinocytes were cultured, the supematant collected and different fractions heated for 10 minutes in a water bath to 60° or 9oOC. Third generation BSM segments (12 dogs) were dissected free of parenchyma. DE by use of a cotton swab and then suspended by a strain gauge in an organ bath. MKSS (800 mcl) abolished the increased ACh sensitivity of the DE BSM (see below). Heating to 600 and 90°C had no significant effect on its relaxing effect. Mean ECmfkD 8.1 X 10-S DE 3.6 X IO4 (~~0.05 relative to DE) DE + MKSS 3.4 x 10-4 DE + MKSS (60°C) 5.9 x 10-4 DE + MKSS (90°C) The slopes of the dose-response curves did not significantly differ when compared by regression analysis. MKSS produces a non-prostanoid canine BSM relaxing factor that is heat-resistant to temperatures up to 9OOC.
SMOOTHMUSCLE (BSM1. N. DAJAM,
ANOWSJU&,F. LOTampa, Florida The in vitro removal of the epithelium from canine and human bronchial airways increases the contractile response of BSM to acetylcholine (ACh). This occurs in dog airways because of the loss of epithehum-derived relaxing factor (EDRF). To determine if other cell types could produce an EDRF-like factor, the effects of MKSS on ACh-induced epithelialized (E) and de-epithelialized (DE) canine BSM contraction were determined. Skin from adult Balb/c mice was treated with trypsin, and the keratinocytes were cultured for 48 hrs at 37O C in Medium 199 supplemented with fetal calf serum and indomethacin. Two pairs of 3rd generation BSM segments from 16 dogs were dissected free of parenchyma; 1 pair was DE by use of a cotton swab. Each segment was suspended by a strain gauge in a 10 ml organ bath to measure tension. ACh induced greater tensions in the DE than E segments (see below). MKSS (800 mcl) abolished the increased ACh sensitivity of the DE BSM but had no effect on the E segments. E E E+MKSS DE+ MKSS mean EC‘50 (M) 2.0X10e5 3.5X10-6’ 2.5X10e5 1.7X10-5” * p <0.0004 relative to E by unpaired t-test ** p co.01 relative to DE by paired t-test The slopes of the DE and DE + MKSS curves did not significantly differ when compared by regression analysis. Culture medium (control) had no effect on E or DE contraction. MKSS produces a non-prostanoid factor that reduces ACh-induced contraction of DE canine BSM. The relationship of this factor to EDRF is unknown.
6aHISTAMINE
Fink.
VA - Milwaukee, WI. We studied the pulmonary alveolar to capillary transport of FI’I’C-D (average m.w. 4400) in isolated perfused rabbit lungs in which the only route for absorption is via the broneboalveolar epithelium into the pulmonary circulation. The lungs were perfused with Krebs Ringer bicarbonate solution containing 5% Bovine Serum Albumin at a flow rata of 200 ml/min, and were ventilated with a constant volume piston ventilator. One ml of water eontaining 1 mg FITC-D with or without 25 pg/ml of H was instilled into the airways. The fraction of instilled FITCD that reached the perfusate with time was determined spectrofluorometrically. Inclusion of H in the instillate resulted in a significantly increased rate of dextran absorption from the bronchoalveolar surface into the perfusate (figure). The calculated t If2 for FITC-D absorption was 125 + 12 BE min in the control and 33 f 5 SE min with the H, (p < 0.001). The results indicate that the measurement of the absorption of instilled FITC-D through isolated rabbit lungs is a reproducible and useful method for evaluating lung alveolar epithelial permeability. H can increase this permeability.
Abstracts
87 1, PART 2
67
65
155
E CANINE BRONCU
Phil&lphi8Jk Todetermineif antigen&E intemctionr cm induce prolonged bosophil histaminereleaae@Wve studiedthe tem00raloatterneof %BfiRin 10a&mice8fter serial and&n,d-10 pm/ml) incubations &rr 4 hours (Table1) TIHEWURS) ;L 1 2 llt2 7tl 6al 39iaf 11*212*2= i.1 6Oi7' 1 16t3* a*2 l&4* 8t2 ~0p~0.01vsOPSN2Uf~p~005vsOPNU
fllmsi
9 9t2 7il lOi 7i2
Whilethe maimurn BkRoccukedin the 1sthr significant increasesof BERoccurredafter the 1sthr at eachof the antigen doses.Wealsocompued the sum of the BHRafter antigen in : ahhe 1sthr $1 m undisturbed fashion from 2-5hrs md c) serial removalandrexposure to fresh antigen(2,3,4hrs), Table2. TABLE2. TM (HOURS) 1 lOt2 39t9' 59t7' :0 5lt6* l p~O.OlvsOPNU,,=P
F 0.1
lit1 = z 35t6*r' 19t5 19t5 3%t? 19*3* < O.Olvs2-5HR5 Serialreplacement of Wigen al 2.3 and 4 hrs resultedin 37-85X highercumuluive BHRuUnthe2-)hr
undklurbed incubations.Furthermomprolonged ewosun to antigen by either methodresultedin an additional3230%BRRalter the 1 hr incubations. Theseobsemationsmayexplain the pattern of prolonged HRve have previously observed during prolonged antigen exposurein cutaneousallergic reactions.
66
-Cl.
OF v KERATINGCVED RP&AXING FACTOR. w M.D.. D. N, Wtieht. M.D.. D. K. Ledford. M.D.. J. J. Krzanowski, Ph.D.. H. A. Sh&ay&* M.D.. Ph.D.. R. F. Lw Tampa, Florida and ‘Zagazig. Egypt Previous work has shown that the in vitro removal of epithelium from canine bronchial airways increases the hyperresponsiveness of the bronchial smooth muscle (BSM) to acetylcholine (ACh) due to the loss of epithelium-derived relaxing factor. We have previously demonstrated that cultured murine keratinocyte suspension supernatant (MKSS) produces a non-prostanoid factor that reduces the AChinduced contraction of de-epithelialized (DE) canine BSM. The effect of 60° and 90°C on this factor was determined. Murine keratinocytes were cultured, the supematant collected and different fractions heated for 10 minutes in a water bath to 60° or 9oOC. Third generation BSM segments (12 dogs) were dissected free of parenchyma. DE by use of a cotton swab and then suspended by a strain gauge in an organ bath. MKSS (800 mcl) abolished the increased ACh sensitivity of the DE BSM (see below). Heating to 600 and 90°C had no significant effect on its relaxing effect. Mean ECmfkD 8.1 X 10-S DE 3.6 X IO4 (~~0.05 relative to DE) DE + MKSS 3.4 x 10-4 DE + MKSS (60°C) 5.9 x 10-4 DE + MKSS (90°C) The slopes of the dose-response curves did not significantly differ when compared by regression analysis. MKSS produces a non-prostanoid canine BSM relaxing factor that is heat-resistant to temperatures up to 9OOC.
SMOOTHMUSCLE (BSM1. N. DAJAM,
ANOWSJU&,F. LOTampa, Florida The in vitro removal of the epithelium from canine and human bronchial airways increases the contractile response of BSM to acetylcholine (ACh). This occurs in dog airways because of the loss of epithehum-derived relaxing factor (EDRF). To determine if other cell types could produce an EDRF-like factor, the effects of MKSS on ACh-induced epithelialized (E) and de-epithelialized (DE) canine BSM contraction were determined. Skin from adult Balb/c mice was treated with trypsin, and the keratinocytes were cultured for 48 hrs at 37O C in Medium 199 supplemented with fetal calf serum and indomethacin. Two pairs of 3rd generation BSM segments from 16 dogs were dissected free of parenchyma; 1 pair was DE by use of a cotton swab. Each segment was suspended by a strain gauge in a 10 ml organ bath to measure tension. ACh induced greater tensions in the DE than E segments (see below). MKSS (800 mcl) abolished the increased ACh sensitivity of the DE BSM but had no effect on the E segments. E E E+MKSS DE+ MKSS mean EC‘50 (M) 2.0X10e5 3.5X10-6’ 2.5X10e5 1.7X10-5” * p <0.0004 relative to E by unpaired t-test ** p co.01 relative to DE by paired t-test The slopes of the DE and DE + MKSS curves did not significantly differ when compared by regression analysis. Culture medium (control) had no effect on E or DE contraction. MKSS produces a non-prostanoid factor that reduces ACh-induced contraction of DE canine BSM. The relationship of this factor to EDRF is unknown.
6aHISTAMINE
Fink.
VA - Milwaukee, WI. We studied the pulmonary alveolar to capillary transport of FI’I’C-D (average m.w. 4400) in isolated perfused rabbit lungs in which the only route for absorption is via the broneboalveolar epithelium into the pulmonary circulation. The lungs were perfused with Krebs Ringer bicarbonate solution containing 5% Bovine Serum Albumin at a flow rata of 200 ml/min, and were ventilated with a constant volume piston ventilator. One ml of water eontaining 1 mg FITC-D with or without 25 pg/ml of H was instilled into the airways. The fraction of instilled FITCD that reached the perfusate with time was determined spectrofluorometrically. Inclusion of H in the instillate resulted in a significantly increased rate of dextran absorption from the bronchoalveolar surface into the perfusate (figure). The calculated t If2 for FITC-D absorption was 125 + 12 BE min in the control and 33 f 5 SE min with the H, (p < 0.001). The results indicate that the measurement of the absorption of instilled FITC-D through isolated rabbit lungs is a reproducible and useful method for evaluating lung alveolar epithelial permeability. H can increase this permeability.