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Complement activation by the occupational allergen trimellitic anhydride (TMA)
075- COMPLEMENT ACTIVATION BY THE OCCUPATIONAL ALLERGEN TRIMELLITIC ANHYDRIDE (TMA). J. F. Regal, C. P. Larsen, T. M. Kane. Dept of Pharmacology, University of Minnesota, Duluth, MN 55812 USA. Guinea pigs, actively sensitized with TMA and challenged intratracheally with TMA conjugated to guinea pig serum albumin (TMA-GPSA), respond with eosinophil, neutrophil and mononuclear cell infiltration into the lungs and an increase in C3a in the bronchoalveolar lavage. Cobra venom factor was previously shown to attenuate this TMAinduced cell intiltration (J Pharmacol Exp Ther 273:793-801. 1995). To determine if TMA-GPSA could cause complement activation in vitro, TMA-GPSA and GPSA were incubated at 37 C for 15 min with complement from either normal unsensitized guinea pigs or guinea pigs sensitized intradermally with TMA. Complement activation was assessed by measuring C3 conversion products by immunofixation techniques. In the absence of specific antibody, TMA-GPSA caused significant C3 conversion both in normal and in C4 deficient serum. In the presence of specific antibody, even greater TMA-GPSA induced C3 conversion was demonstrated and this conversion was completely prevented by MgEGTA treatment. Thus, TMAGPSA causes complement activation by the classical pathway in the presence of specific antibody. However, even without specific antibody, TMA-GPSA induces significant C3 conversion via the alternative pathway, suggesting that antibody independent complement activation is important in TMA induced respiratory hypersensitivity in the guinea pig.
076- EVALUATING A ROLE OF COMPLEMENT ACI’IVATION IN ALLERGICLUNG DAMAGE INDUCED BY REPEATED INHALATION OF ANTIGBN Masayoshl Abe, *Naomi Shim+! *Kamhlko Shibata, **Norlyuki Sakata, Takshi Katsumgi, ***Hiroyasu Akatsu, ***Hickchika Oka& Dept. Phamtmol., *L&o. Biodyn. “‘2nd Pathol., Sch. Med, Fukuoka Univ., Fukuoka 814-0180. ***Dept. Mol. Biol.. Nagoya City Univ. Sch. Med., Nagoya, 467-8601. Japan. We tried to make a model mimicking chronic broncltlal mthma and to evaluate a role of complement activation in this model. Male B-N rats wereactively sensitized by injecting ovalhumln (OA). The sensitized rats receivedrepeatedinhalation of OAaml there&a airway resistance (Raw) was estimated by measuring &low and esophageal pitssue under urethane anesthesia and artificial ventilation. In o&r to examine histological findngs, lungs were obtained 6 h after the intratmcheal challenge of OA. Intmtrmheal mkttinistmtion of OA into the sensitized rats resulted in two phases of airway tesponse (AR), immediate (IAR) and late airway response (LAR). While IAR tendedto decmasein proportion to times of OA inhalation, LAR turned to be pmminent. Pretreatment with soluble complement teceptor type 1 (sCR1) inhibited both teaponses. On the other hand, the repeated inhalation of OA resulted in inflammatory cell infiltration around airway includng patchy gmuulomatous lesion and perivascular e&ma The pmtreatment with sCR1 inhibited these histological Endings in lungs. Western and Northern blottings for C5aR and C3aR showed upregulated expression in lungs after the repeatedinhalation of OA. These results suggest that complement activation plays a significant role in allergic lung damage imktced by repeated inhalation of antigen.
077- COMPARATIVE EXPRESSION OF ANAPHYLATOXIN C3A AND C5A RECEPTORS ON LUNG BRONCHJAL EPITHELIAL AND SMOOTH MUSCLE CELLS IN MOUSE MODELS OF ASTHMA AND SEPSIS Scott. M. Drouint, Jens Kildsgaard’, Joie Haviland’, Hong Pen fiaa, Paul B. McCrays, Btiau F. Tacks, and Rick A. Wet& ‘Uttiversity ofTexas-Houston Institute ofMolecular Medicine and *Univeralty of Iowa College ofMedicine, USA The presetme of C3a and C5a inthe lung results in respiratory distress characterized by leukocyte recruitment and bronchial smooth muscle consbiotlon. Although the C3a aud CSa ra+tors could potentially mediate these inflammatoty effects, previous studlee have only documented the expression of the CSa receptor on bronchial epiiheiial cells. Distribution of the C3a recqtor in the lung relative to the C5a recqtor, is currently unknown. In the present study, we have compared the expression of the C3a and C5a mceptom in the hmg under normal and inflammatory conditions. Analysis by immunobistochemistry and in situ hybridiiation revealed expression of C3a and C5a mceptor protein and mRNA on the bronchioles of mouse and human lung. In normal human lung, C3a and C5a receptor expression was detected on bronchial epithelial and smooth muscle calls. Receptor expression was also comp2ued in mouse models of sepsis and asthma. In mice stimulated with LB, expression of both receptors increased on the bronchioles relative to saline controls. However, in the asthma modal, only increases in C3a receptor expresalon was detected on lung bnmchloles. Further analysis revealed the expression of C3a and C5a mccptom on primary cultures of human brottchlal epithelial cells and antooth muscle cells. These results demonstrate C3a and C5a receptor expression in cells indigenous to the lung and suggest a mle for these receptors during lung inflammation.
078- Should we use probioties as supportive treatment for hereditary angioedema? Kazimierz Madalinski, Hanna Jaworska, Hanna Gregorek Department of Clinical Immunology, Children’s Memorial Health Institute, 04-730 Warsaw, Poland Antilibrinolitic agents, such as aminocaproic and tranexamic acid, and attenuated androgens, like danazol and stanazolol, are commonly used for prophylaxis of hereditary or acquired angioedema. Prolonged use of these drugs, which is often needed, may result in severe side effects from gastrointestinal, respiratory, genitourinary tracts, skin, and from endocrine and nervous systems. The most striking was the development of hepatocellular adenoma or carcinoma after 15 yrs of treatment. We have checked the practical use of prohiotics, natural inductors of IFN-y, which may upregulate Cl-inh synthesis. Here we present the fust results from the preliminary therapeutic study undertaken on 10 patients with HAE. Patients ate regularly 500 g/ daily of yoghurt containing _L.acidophilus during 6 months, while maintaining the basic treatment. C4 and Cl-inh were measured by nephelometry and radial immunodifusion and CH50 by the modified Meyer’s method. Clinical symptoms were evaluated according to an arbitary scale of severity of attacks: 6-severe, 4medium, 2-mild, O-nil. We have compared the tested parameters 01 patients before and after the yoghurt treatment. During the study period the mean score of clinical symptoms has decreased from 4.7 to 3.3 (30%); p< 0.003; the value of C4 component has risen from 4.51 to 7.62 mg/dL (rise of 69%); p
076- EVALUATING A ROLE OF COMPLEMENT ACI’IVATION IN ALLERGICLUNG DAMAGE INDUCED BY REPEATED INHALATION OF ANTIGBN Masayoshl Abe, *Naomi Shim+! *Kamhlko Shibata, **Norlyuki Sakata, Takshi Katsumgi, ***Hiroyasu Akatsu, ***Hickchika Oka& Dept. Phamtmol., *L&o. Biodyn. “‘2nd Pathol., Sch. Med, Fukuoka Univ., Fukuoka 814-0180. ***Dept. Mol. Biol.. Nagoya City Univ. Sch. Med., Nagoya, 467-8601. Japan. We tried to make a model mimicking chronic broncltlal mthma and to evaluate a role of complement activation in this model. Male B-N rats wereactively sensitized by injecting ovalhumln (OA). The sensitized rats receivedrepeatedinhalation of OAaml there&a airway resistance (Raw) was estimated by measuring &low and esophageal pitssue under urethane anesthesia and artificial ventilation. In o&r to examine histological findngs, lungs were obtained 6 h after the intratmcheal challenge of OA. Intmtrmheal mkttinistmtion of OA into the sensitized rats resulted in two phases of airway tesponse (AR), immediate (IAR) and late airway response (LAR). While IAR tendedto decmasein proportion to times of OA inhalation, LAR turned to be pmminent. Pretreatment with soluble complement teceptor type 1 (sCR1) inhibited both teaponses. On the other hand, the repeated inhalation of OA resulted in inflammatory cell infiltration around airway includng patchy gmuulomatous lesion and perivascular e&ma The pmtreatment with sCR1 inhibited these histological Endings in lungs. Western and Northern blottings for C5aR and C3aR showed upregulated expression in lungs after the repeatedinhalation of OA. These results suggest that complement activation plays a significant role in allergic lung damage imktced by repeated inhalation of antigen.
077- COMPARATIVE EXPRESSION OF ANAPHYLATOXIN C3A AND C5A RECEPTORS ON LUNG BRONCHJAL EPITHELIAL AND SMOOTH MUSCLE CELLS IN MOUSE MODELS OF ASTHMA AND SEPSIS Scott. M. Drouint, Jens Kildsgaard’, Joie Haviland’, Hong Pen fiaa, Paul B. McCrays, Btiau F. Tacks, and Rick A. Wet& ‘Uttiversity ofTexas-Houston Institute ofMolecular Medicine and *Univeralty of Iowa College ofMedicine, USA The presetme of C3a and C5a inthe lung results in respiratory distress characterized by leukocyte recruitment and bronchial smooth muscle consbiotlon. Although the C3a aud CSa ra+tors could potentially mediate these inflammatoty effects, previous studlee have only documented the expression of the CSa receptor on bronchial epiiheiial cells. Distribution of the C3a recqtor in the lung relative to the C5a recqtor, is currently unknown. In the present study, we have compared the expression of the C3a and C5a mceptom in the hmg under normal and inflammatory conditions. Analysis by immunobistochemistry and in situ hybridiiation revealed expression of C3a and C5a mceptor protein and mRNA on the bronchioles of mouse and human lung. In normal human lung, C3a and C5a receptor expression was detected on bronchial epithelial and smooth muscle calls. Receptor expression was also comp2ued in mouse models of sepsis and asthma. In mice stimulated with LB, expression of both receptors increased on the bronchioles relative to saline controls. However, in the asthma modal, only increases in C3a receptor expresalon was detected on lung bnmchloles. Further analysis revealed the expression of C3a and C5a mccptom on primary cultures of human brottchlal epithelial cells and antooth muscle cells. These results demonstrate C3a and C5a receptor expression in cells indigenous to the lung and suggest a mle for these receptors during lung inflammation.
078- Should we use probioties as supportive treatment for hereditary angioedema? Kazimierz Madalinski, Hanna Jaworska, Hanna Gregorek Department of Clinical Immunology, Children’s Memorial Health Institute, 04-730 Warsaw, Poland Antilibrinolitic agents, such as aminocaproic and tranexamic acid, and attenuated androgens, like danazol and stanazolol, are commonly used for prophylaxis of hereditary or acquired angioedema. Prolonged use of these drugs, which is often needed, may result in severe side effects from gastrointestinal, respiratory, genitourinary tracts, skin, and from endocrine and nervous systems. The most striking was the development of hepatocellular adenoma or carcinoma after 15 yrs of treatment. We have checked the practical use of prohiotics, natural inductors of IFN-y, which may upregulate Cl-inh synthesis. Here we present the fust results from the preliminary therapeutic study undertaken on 10 patients with HAE. Patients ate regularly 500 g/ daily of yoghurt containing _L.acidophilus during 6 months, while maintaining the basic treatment. C4 and Cl-inh were measured by nephelometry and radial immunodifusion and CH50 by the modified Meyer’s method. Clinical symptoms were evaluated according to an arbitary scale of severity of attacks: 6-severe, 4medium, 2-mild, O-nil. We have compared the tested parameters 01 patients before and after the yoghurt treatment. During the study period the mean score of clinical symptoms has decreased from 4.7 to 3.3 (30%); p< 0.003; the value of C4 component has risen from 4.51 to 7.62 mg/dL (rise of 69%); p