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Cotoneaster lacteus: A New Lipid Transfer Protein (LTP) Source from the Rosaceae Family
Abstracts S101
J ALLERGY CLIN IMMUNOL VOLUME 121, NUMBER 2
Hereditary Angioedema: a New Mutation of the C1 Inhibitor Gene in a Brazilian Family M. F. Ferraro1, W. Sarti1, A. C. Souza1, E. A. Donadi1, E. Castelli1, K. Bork2, L. K. Arruda1; 1School of Medicine of Ribeira˜o Preto - USP, Ribeira˜o Preto, BRAZIL, 2Johannes Gutenberg University, Mainz, GERMANY. RATIONALE: To describe clinical features and the underlying molecular defect within a family with hereditary angioedema (HAE). METHODS: History of episodes of angioedema and attacks of abdominal pain was investigated among members of a Brazilian family with an index case of HAE. Family pedigree was constructed with 151 individuals distributed in 5 generations. Fourteen patients were interviewed and examined, and provided information about relatives. Sequencing of the eight coding exons of C1 inhibitor (C1-INH) was carried out in three patients. Levels of C4 were determined by nephelometry and quantification of C1-INH was performed by radial immunodiffusion. RESULTS: Thirty-five individuals (23 women) presented history of recurrent angioedema. Eight patients, among the 14 who underwent detailed evaluation, fulfilled diagnostic criteria for HAE. Six additional symptomatic patients presented normal C1-INH levels and activity, with (n 5 3) or without (n 5 3) low C4 levels. Two patients with diagnosed HAE were heterozygous for a single-nucleotide deletion at position 351 on exon 3 (c.351 del C), which could lead to frameshift and disruption of the coding sequence for amino acid 117, within the serpin domain. In one patient with severe clinical symptoms most likely attributable to HAE and normal C1INH levels, no abnormalities were found on coding exons of the C1-INH gene. CONCLUSIONS: Heterogeneous laboratory features were found in patients with angioedema who are members of a family of individuals with established diagnosis of HAE. We report a new mutation of the C1INH gene, which may be related to low levels of C1-INH leading to symptoms of HAE in a Brazilian family. Funding: School of Medicine of Ribeira˜o Preto
390
Assessing Clinical Value of Elevated IgE Receptor Antibody Titers (aFceRI) in Patients with Chronic Urticaria M. L. Oswalt, T. H. Ly, G. D. Marshall; University of Mississippi Medical Center, Jackson, MS. RATIONALE: aFceRI is present in approximately 30% of patients with chronic urticaria. We examined whether the presence of the antibody was associated with other autoantibody titers and/or could be predictive of subsequent clinical control in our patient population. METHODS: A retrospective chart review of patients with chronic urticaria (708.1,8,9) from a University-based allergy clinic was performed for the presence of aFceRI, total number of medications, immunomodulatory medications, thyroid-peroxidase antibody (aTPO), and presence or absence of clinical control. Patients that required immunomodulatory drugs, presence of lesions at least weekly, and/or interference with normal routine were considered uncontrolled. Two investigators blinded to laboratory tests independently assessed control. If there was a grading disparity for an individual patient, a third investigator reviewed the chart for the final determination. RESULTS: 34 patient charts were identified whose aFceRI data were recorded in their charts. As expected, uncontrolled patients were taking more total medications (p 5 0.033), more immunomodulatory medications (p 5 0.0037), and were more likely to have aTPO (p 5 0.053). However, there were no differences in clinical control between patients with elevated vs normal FceRI antibody levels (p 5 0.2335) and no association of aFceRI with aTPO (p 5 0.7824). CONCLUSIONS: aFceRI titers were not associated with clinical control or autoantibodies in our population. These data question the value of obtaining aFceRI in individual patients with chronic urticaria for therapeutic choices or predicting future treatment response. Prospective studies are needed to define the practical clinical value of this test.
391
Studies of the Mechanisms of Bradykinin Generation in Hereditary Angioedema Plasma K. Joseph, T. B. Tuscano, A. P. Kaplan; Medical University of South Carolina, Charleston, SC. RATIONALE: Factor XII-dependent bradykinin formation is thought to be responsible for the swelling associated with the various forms of C1 inhibitor deficiency while complement activation is augmented during attacks of swelling. We wished to further elucidate those interactions of the kinin-forming cascade that lead to complement activation during attacks of swelling and to determine whether fibrinolysis is augmented as well. METHODS: Spontaneous activation of normal plasma and plasma of patients with Hereditary Angioedema (HAE) was compared by prolonged incubation in 96 well plates (polystyrene) as well as by activation with kaolin. We assayed for activated factor XII, prekallikrein, high molecular weight kininogen cleavage, C4a formation, and plasmin-a2-antiplasmin complex formation. RESULTS: HAE plasma demonstrates augmented factor XII activation, production of factor XII fragment (XIIf), prekallikrein activation and HK cleavage when compared to normal plasma. As a result, bradykinin formation is markedly increased. Production of factor XIIf, demonstrated for the first time in whole plasma, may be responsible for C1 activation based on C4a production. There is also increased baseline levels of C4a and plasmin-a2 antiplasmin complexes in HAE plasma which increase further upon activation with kaolin. CONCLUSIONS: All parameters indicative of activation of the bradykinin-forming cascade are activated in HAE plasma vs normal plasma. The factor XII-dependent fibrinolytic cascade is also activated. Complement activation during attacks of swelling are likely a result of HFf activation of C1r. Therapy with C1 inhibitor (purified) should prevent activation of all factor XII-dependent pathways including bradykinin formation, increased fibrinolytic activity, and activation of the classical complement pathway. Funding: Lev Pharmaceuticals Inc.
392
Cotoneaster lacteus: A New Lipid Transfer Protein (LTP) Source from the Rosaceae Family J. Torres1, C. Escudero1, D. Iba´n˜ez1, M. Lombardero2, A. Ledesma2, C. ˜O Mun˜oz1, T. Laso1; 1HOSPITAL INFANTIL UNIVERSITARIO NIN ´ S, MADRID, SPAIN, 2R&D Department. ALK-Abello´ S.A., JESU MADRID, SPAIN. RATIONALE: LTP are the only allergens identified on Rosaceae fruit allergic patients without pollinosis. A 9-year-old girl, peach-allergic and without pollinosis, presented a contact urticaria due to some ’’red fruits’’ (RF) she has been touching in a park. METHODS: Skin prick tests (SPT) with a pollen standard kit and with palm profilin (ALK-Abello´, SA. Madrid, Spain) were performed. Prickprick with some Rosaceae fruits and with the RF were performed on the patient and on 4-non-pollinic peach-allergic patients used as controls. An SDS-PAGE with RF extract and IgE-immunoblotting was carried out. In addition, immunodetection using a polyclonal anti-Pru p3 serum was also performed. Specific IgE to Pru p3 (peach LTP) and Mal d4 (apple profilin) were determined using the ADVIA-CentaurÒ-platform. RESULTS: The RF was identified as Cotoneaster lacteus (C.l.), species that belongs to the Rosaceae family. SPT were negative to pollen and to profilin. Prick-prick were positive with RF and with the other fruits tested on the patient and on 3 controls. A 14-15 and a 9-10 kDa IgE-binding bands were found. A homologous protein to Pru p3 was detected in the RF extract, by immunodetection with the anti-Pru p3 polyclonal serum. All patients had specific IgE to peach LTP but were negative to profilin. CONCLUSIONS: Immunological study suggests that LTP from Cotoneaster lacteus RF was responsible for the contact urticaria. We have to consider in our practice the possibility that Rosaceae fruit allergic patients could have an allergic reaction when manipulating the C.I. ’’red fruit’’, species widely extended in parks and gardens.
SUNDAY
389
J ALLERGY CLIN IMMUNOL VOLUME 121, NUMBER 2
Hereditary Angioedema: a New Mutation of the C1 Inhibitor Gene in a Brazilian Family M. F. Ferraro1, W. Sarti1, A. C. Souza1, E. A. Donadi1, E. Castelli1, K. Bork2, L. K. Arruda1; 1School of Medicine of Ribeira˜o Preto - USP, Ribeira˜o Preto, BRAZIL, 2Johannes Gutenberg University, Mainz, GERMANY. RATIONALE: To describe clinical features and the underlying molecular defect within a family with hereditary angioedema (HAE). METHODS: History of episodes of angioedema and attacks of abdominal pain was investigated among members of a Brazilian family with an index case of HAE. Family pedigree was constructed with 151 individuals distributed in 5 generations. Fourteen patients were interviewed and examined, and provided information about relatives. Sequencing of the eight coding exons of C1 inhibitor (C1-INH) was carried out in three patients. Levels of C4 were determined by nephelometry and quantification of C1-INH was performed by radial immunodiffusion. RESULTS: Thirty-five individuals (23 women) presented history of recurrent angioedema. Eight patients, among the 14 who underwent detailed evaluation, fulfilled diagnostic criteria for HAE. Six additional symptomatic patients presented normal C1-INH levels and activity, with (n 5 3) or without (n 5 3) low C4 levels. Two patients with diagnosed HAE were heterozygous for a single-nucleotide deletion at position 351 on exon 3 (c.351 del C), which could lead to frameshift and disruption of the coding sequence for amino acid 117, within the serpin domain. In one patient with severe clinical symptoms most likely attributable to HAE and normal C1INH levels, no abnormalities were found on coding exons of the C1-INH gene. CONCLUSIONS: Heterogeneous laboratory features were found in patients with angioedema who are members of a family of individuals with established diagnosis of HAE. We report a new mutation of the C1INH gene, which may be related to low levels of C1-INH leading to symptoms of HAE in a Brazilian family. Funding: School of Medicine of Ribeira˜o Preto
390
Assessing Clinical Value of Elevated IgE Receptor Antibody Titers (aFceRI) in Patients with Chronic Urticaria M. L. Oswalt, T. H. Ly, G. D. Marshall; University of Mississippi Medical Center, Jackson, MS. RATIONALE: aFceRI is present in approximately 30% of patients with chronic urticaria. We examined whether the presence of the antibody was associated with other autoantibody titers and/or could be predictive of subsequent clinical control in our patient population. METHODS: A retrospective chart review of patients with chronic urticaria (708.1,8,9) from a University-based allergy clinic was performed for the presence of aFceRI, total number of medications, immunomodulatory medications, thyroid-peroxidase antibody (aTPO), and presence or absence of clinical control. Patients that required immunomodulatory drugs, presence of lesions at least weekly, and/or interference with normal routine were considered uncontrolled. Two investigators blinded to laboratory tests independently assessed control. If there was a grading disparity for an individual patient, a third investigator reviewed the chart for the final determination. RESULTS: 34 patient charts were identified whose aFceRI data were recorded in their charts. As expected, uncontrolled patients were taking more total medications (p 5 0.033), more immunomodulatory medications (p 5 0.0037), and were more likely to have aTPO (p 5 0.053). However, there were no differences in clinical control between patients with elevated vs normal FceRI antibody levels (p 5 0.2335) and no association of aFceRI with aTPO (p 5 0.7824). CONCLUSIONS: aFceRI titers were not associated with clinical control or autoantibodies in our population. These data question the value of obtaining aFceRI in individual patients with chronic urticaria for therapeutic choices or predicting future treatment response. Prospective studies are needed to define the practical clinical value of this test.
391
Studies of the Mechanisms of Bradykinin Generation in Hereditary Angioedema Plasma K. Joseph, T. B. Tuscano, A. P. Kaplan; Medical University of South Carolina, Charleston, SC. RATIONALE: Factor XII-dependent bradykinin formation is thought to be responsible for the swelling associated with the various forms of C1 inhibitor deficiency while complement activation is augmented during attacks of swelling. We wished to further elucidate those interactions of the kinin-forming cascade that lead to complement activation during attacks of swelling and to determine whether fibrinolysis is augmented as well. METHODS: Spontaneous activation of normal plasma and plasma of patients with Hereditary Angioedema (HAE) was compared by prolonged incubation in 96 well plates (polystyrene) as well as by activation with kaolin. We assayed for activated factor XII, prekallikrein, high molecular weight kininogen cleavage, C4a formation, and plasmin-a2-antiplasmin complex formation. RESULTS: HAE plasma demonstrates augmented factor XII activation, production of factor XII fragment (XIIf), prekallikrein activation and HK cleavage when compared to normal plasma. As a result, bradykinin formation is markedly increased. Production of factor XIIf, demonstrated for the first time in whole plasma, may be responsible for C1 activation based on C4a production. There is also increased baseline levels of C4a and plasmin-a2 antiplasmin complexes in HAE plasma which increase further upon activation with kaolin. CONCLUSIONS: All parameters indicative of activation of the bradykinin-forming cascade are activated in HAE plasma vs normal plasma. The factor XII-dependent fibrinolytic cascade is also activated. Complement activation during attacks of swelling are likely a result of HFf activation of C1r. Therapy with C1 inhibitor (purified) should prevent activation of all factor XII-dependent pathways including bradykinin formation, increased fibrinolytic activity, and activation of the classical complement pathway. Funding: Lev Pharmaceuticals Inc.
392
Cotoneaster lacteus: A New Lipid Transfer Protein (LTP) Source from the Rosaceae Family J. Torres1, C. Escudero1, D. Iba´n˜ez1, M. Lombardero2, A. Ledesma2, C. ˜O Mun˜oz1, T. Laso1; 1HOSPITAL INFANTIL UNIVERSITARIO NIN ´ S, MADRID, SPAIN, 2R&D Department. ALK-Abello´ S.A., JESU MADRID, SPAIN. RATIONALE: LTP are the only allergens identified on Rosaceae fruit allergic patients without pollinosis. A 9-year-old girl, peach-allergic and without pollinosis, presented a contact urticaria due to some ’’red fruits’’ (RF) she has been touching in a park. METHODS: Skin prick tests (SPT) with a pollen standard kit and with palm profilin (ALK-Abello´, SA. Madrid, Spain) were performed. Prickprick with some Rosaceae fruits and with the RF were performed on the patient and on 4-non-pollinic peach-allergic patients used as controls. An SDS-PAGE with RF extract and IgE-immunoblotting was carried out. In addition, immunodetection using a polyclonal anti-Pru p3 serum was also performed. Specific IgE to Pru p3 (peach LTP) and Mal d4 (apple profilin) were determined using the ADVIA-CentaurÒ-platform. RESULTS: The RF was identified as Cotoneaster lacteus (C.l.), species that belongs to the Rosaceae family. SPT were negative to pollen and to profilin. Prick-prick were positive with RF and with the other fruits tested on the patient and on 3 controls. A 14-15 and a 9-10 kDa IgE-binding bands were found. A homologous protein to Pru p3 was detected in the RF extract, by immunodetection with the anti-Pru p3 polyclonal serum. All patients had specific IgE to peach LTP but were negative to profilin. CONCLUSIONS: Immunological study suggests that LTP from Cotoneaster lacteus RF was responsible for the contact urticaria. We have to consider in our practice the possibility that Rosaceae fruit allergic patients could have an allergic reaction when manipulating the C.I. ’’red fruit’’, species widely extended in parks and gardens.
SUNDAY
389