This action might not be possible to undo. Are you sure you want to continue?
Andre Talvani£, Manoel O.C. Rocha§, Antonio L. Ribeiro§, Enri Borda∂, Leonor Sterin-Borda∂ & Mauro M Teixeira£.
Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas and, §Pós-
Graduação em Medicina Tropical, Departamento de Clínica Médica, Faculdade de Medicina, UFMG,Belo Horizonte, Brasil and ∂Pharmacology Unit, School of Dentistry, University of Buenos Aires, Buenos Aires, Argentina.
Short title: Anti-M2 autoantibodies in Chagas Heart Disease
Footnote page: (1) Informed consent was obtained from all patients and non-infected individuals.
Human experimental guidelines of the Brazilian Ministry of Health were followed in the conduct of the experiments described here. The authors have not commercial or other association that may pose a conflict of interest.
This investigation received financial support from FAPEMIG, the UNDP/World
Bank/WHO Special Programme for Research and Training in Tropical Diseases (TDRWHO A970728) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq-Brazil) and National Research Council (CONICET) of Argentina, TDR-WHO A20771 and University of Buenos Aires (UBACYT), Buenos Aires, Argentina.
Address for correspondence and reprints request
Mauro M Teixeira, M.D. Departamento de Bioquímica e Imunologia Instituto de Ciências Biológicas Universidade Federal de Minas Gerais Av. Antônio Carlos, 6627 - Pampulha 31270-901 BELO HORIZONTE MG BRASIL Phone # 55 31 3499 2651 (direct line) Fax # 55 31 3441 5963 e-mail: firstname.lastname@example.org
Alternative contact: Dr Leonor Sterin-Borda (email@example.com).
Patients with apical lesion. The levels of anti-M2 antibodies were lower in patients with the indeterminate form than patients with CCC. Methods: Anti-M2 cholinergic and anti-β1 adrenergic autoantibodies were measured in sera of non-infected subjects and chagasic patients with the various clinical forms of the disease using ELISA. with exercise-induced ventricular arrhythmias. ventricular arrhythmias and chronotropic incompetence. levels of both antibodies were similar in patients with CCC. Conclusion: The levels of anti-neurotransmitter autoantibodies in serum of patients with Chagas heart disease seemed not to correlate with the severity of the left ventricular dysfunction.Abstract Background: Anti-neurotransmitter receptor autoantibodies are found in the serum of chagasic patients and may play a role in the pathophysiology of the disease. In the present study. Key Words: chagas cardiomyopathy – chagas disease – neurotransmitter receptor antibodies – anti-adrenergic antibodies – anti-cholinergic antibodies 3 . This is consistent with lack of correlation between levels of autoantibodies and various parameters of ventricular dysfunction. Results: The optical density (OD) and the frequency of anti-M2 and anti-β1 antibodies were higher in chagasic patients than in non-infected subjects. suggesting that anti-M2 cholinergic autoantibodies may play a role in the pathogenesis of chagasic cardiac dysautonomia. the levels of anti-M2 autoantibodies are increased in patients in which apical aneurysm. or those with chronotropic insufficiency had higher levels of anti-M2 autoantibodies. regardless of severity. we investigated the presence of autoantibodies and their correlation with the severity of chronic chagasic cardiomyopathy (CCC). Nevertheless.
In a significant proportion of the latter patients. Although a strong association between circulating antipeptide M2 muscarinic acetylcholine receptor (mAChR) autoantibodies and the presence of patients' low heart rate variability index. cruzi). severe heart disease occurs and is frequently the cause of death. There are several hypotheses that try to explain the pathogenesis of severe heart disease in infected individuals. including the role of parasite persistence (1.2). bradycardia and cardiac or esophageal autonomic dysfunction in chronic chagasic patients was verified (15). the presence and levels of anti-adrenergic (β1) and anti-muscarinic (M2) receptor antibodies were evaluated in serum of a group of 58 individuals. Here. chronic chagasic cardiomyopathy (CCC) affects around 30% of individuals infected with the protozoan parasite Trypanosoma cruzi (T.Introduction In Latin America. It has been argued that the blockade of neurotransmitter receptors by anti-receptor antibodies contributes to the autonomic dysfunction and worse clinical evolution (12-14). 4 . Chronic chagasic cardiomyopathy is characterized mainly by a dilated cardiomyopathy complicated by frequent and complex ventricular arrhythmias and/or conduction defects (8). autoimmune events (3-6) and microvascular dysfunction (7). Autonomic dysfunction occurs early in the course of the disease and may associate with worse prognosis (9-12). it is not known whether the presence and/or titer of anti-receptor antibodies correlate with CCC severity. We also investigated the correlation between the titers of autoantibodies and the following aspects of clinical CCC manifestations: left ventricular systolic function and response to effort.
free T4.g. creatinine. potassium. blood urea nitrogen. a 24 hour Holter examination.g. patients with indeterminate form (IND) (n=8) or chronic chagasic cardiomyopathy grade I (CCC I) (n=8) were those with normal ECG and radiological studies or with only minor alterations in their ECHO (e. TSH. diabetes. chest X-Ray. respectively. Chagasic patients were also categorized into groups according to the degree of heart dysfunction. electrocardiogram (ECG). Patients with hypertension. All individuals were recruited at the Referral Center for Training on Infectious and Parasitic Diseases (CTR-DIP) at Hospital das Clinicas. thyroid or renal disturbances or any other cardiac or systemic diseases and those using steroidal drugs were excluded from this study. Patients classified as CCC II/III (n=7) were those with minor or moderate ECG alterations. Universidade Federal de Minas Gerais (UFMG) and underwent a complete clinical examination and performed the following laboratory workup: full blood count. Briefly.Methods Study population We performed this initial study with 6 healthy individuals and 52 chagasic patients with different clinical forms of the disease. right bundle branch block or uniform ventricular premature contractions. including block of the anterosuperior division of the left branch. as these conditions could prevent adequate interpretation of cardiac disease severity on immune parameters. as previously described (8). left bundle branch block. echoDopplercardiography (ECHO) and a treadmill exercise test. left anterior divisional block with right 5 . regional contraction defects). Patients classified as CCC IV (n=15) were those manifesting severe conduction defects (e. The study received ethical clearance from the Ethics Review Board of Universidade Federal de Minas Gerais. glucose.
centrifuged and stored at -80oC until used in an immunoassay (ELISA) with M2 synthetic cholinergic peptide (18) and β1 adrenergic synthetic peptide (14) as coating antigens. The left ventricular ejection fraction (LVEF) was obtained by Simpson's method using the software provided with the equipment. irrespective of the presence or not of arrhythmias or conduction defects (8). Chronotropic insufficiency was arbitrarily defined as the inability to achieve at least 85% of the predicted heart rate according to Astrand’s formula (220-age) at peak exercise (16). Patients underwent ECHO with color flow using an ATL Philips HDI 5000 apparatus operated by an experienced echocardiographer. non-sustained or sustained ventricular tachycardia). Patients with exercise-induced arrhythmias were those who presented non-sustained ventricular tachycardia or increase ventricular premature beats number (NVPB) or complexity (polimorphic or repetitive forms) clearly related to the exercise. Finally. as observed on the ECHO. A maximal stress test was performed according to the standard Bruce protocol. as previously 6 . Segmental contractility was evaluated according to the method described by American Society of Echocardiography (17) and typical apical lesions in the left ventricle wall were recognized. The control group was formed by non-infected (NI) healthy individuals (Table 1). patients classified as CCC V were those with ventricular enlargement.bundle branch block or total atrioventricular block) or complex ventricular arrhythmias (complex ventricular premature beats. Measurement of antibodies against anti-M2 cholinergic and anti-β1 adrenergic receptors Serum samples were obtained by conventional venipuncture. blinded to the clinical status of the patients.
The samples were assayed in parallel at a 1/50 dilution and optical density (OD) values were measured with an ELISA reader (Uniskan Laboratory System). 7 .05. San Diego.described (19). Analysis was performed using the computer program GraphPrism (GraphPad. For association between variables. CA. Probability values were considered significant when p<0. Comparison between groups carried was out by using Analysis of Variance (ANOVA) followed by Student-Newman-Keuls´s post test (parametric distribution) or Kruskal Wallis followed by Dunn´s post test (non-parametric distribution). This antibody dilution was found to be optimal to separate chronic chagasic cardiomyopathy patients from control indeterminate form of Chagas’ heart disease. USA). data were analyzed using linear regression analysis and Spearman´s rank correlation test. Statistical analysis Data are expressed as means ± SEM or median and interquartile range.
when chagasic patients were grouped according to disease severity (8). It can be seen that the frequency of anti-M2 cholinergic autoantibody was higher in CCC patients (mean values: 86. However. patients with CCC V had lower left ventricle ejection fraction and greater diastolic diameter in comparison with patients with the other degrees of CCC. When the distribution of anti-β1 adrenergic autoantibodies was evaluated no differences in the frequency between CCC patients (mean value 67. The lack of correlation between levels of autoantibodies and disease severity is further re-enforced 8 . There was great variation in the number of ventricular premature beats in 24 hour and patients with CCC II/III or worse had greater number of premature beats than those with IND form or CCC 1 (Table 1).1%) was observed. The non-infected individuals were negative in the study system. Table 2 also shows the distribution of both autoantibodies in the different degrees of CCC and it can be appreciated that no differences existed between the different degrees of CCC. It can be seen that chagasic patients presented greater levels of both autoantibodies.3%) than in the IND form (mean values 37.Results There was no significant difference in the age distribution of non-infected individuals and chagasic patients (Table 1). In agreement with the clinical parameters used to classify the group. there was no difference in the OD values of both M2 and β1 autoantibodies among the different clinical groups (Figure 1).2%) and IND (mean value 77.5%) of Chagas’ disease. The distribution of anti-M2 cholinergic and anti-β1 adrenergic autoantibodies detected by ELISA is shown in Table 2. Figure 1 shows the levels of cholinergic (M2) and adrenergic (β1) autoantibodies in serum from non-chagasic and chagasic individuals.
Moreover. 9 . the levels of anti-M2 antibodies were higher in patients with than those without chronotropic insufficiency (Figure 3B).when the levels of antibodies and clinical parameters of ventricular dysfunction were compared (Table 3). patients presenting apical lesion in the left ventricle had higher levels of anti-M2 antibodies (Figure 2). Patients that presented ventricular arrhythmias during or immediately after exercise had higher levels of anti-M2 antibodies. but not anti-β1 antibodies than patients who did not present exercise-induced arrhythmias (Figure 3A). but not anti-β1 antibodies than patients without apical lesion (Figure 2). Interestingly. The levels of anti-ß1 antibodies were similar in patients with or without chronotropic incompetence (Figure 3B).
Discussion Global systolic left ventricular dysfunction is the strongest predictor of morbidity and mortality during Chagas’ heart disease (20. it is clear that chagasic patients have anti-M2 or anti-ß1 receptor antibodies and that the binding of these autoantibodies to the receptors may have functional consequences. For example. Antibodies against adrenergic and cholinergic receptors are among the many autoantibodies that have been described in Chagas disease. levels of this autoantibody were not able to differentiate the various forms of Chagas heart disease. 10 .14). suggesting that the anti-M2 autoantibody could be used as an early marker of evolution in Chagas cardiomyopathy.12. anti-M2 and anti-β1 antibodies are also found in serum of patients with other forms of heart disease (12. The frequency of anti-M2 autoantibody was higher in CCC than in IND form of the disease. enhance or decrease contractility) and may also interact with the respective receptor and induce sequestration and endocytosis of the receptor (2225). These antibodies may induce acute functional alterations of isolated hearts from experimental animals (e. In regard to the latter possibility. it is unclear whether the autoantibodies have a role in the pathogenesis of Chagas’ disease and/or reflect structural damage to the heart.26-28). It has been argued that an autoimmune response against antigens present in heart tissue may favor the development of the more severe forms of Chagas cardiomyopathy. Our results showed that individuals with chronic chagasic cardiomyopathy had elevated levels of both adrenergic and cholinergic autoantibodies when compared with non-infected controls.21). However. However. antibodies against β1-adrenoceptores and M2 mAChR have been found in the sera of patients and experimental animals with Chagas disease (5.g. Thus.
on the other hand. although also observed in other types of heart disease. exercise-induced arrhythmia and the chronotropic response to exercise. Thus. such as ventricular extra-systoles and conduction abnormalities. Alternatively. and impairment of the 11 . Segmental changes of myocardial contractility. with thinning and deficit preferentially localized on the apex of the left ventricle. the antibodies may not have a direct role in the pathogenesis of the left ventricular dysfunction that accompanies the most severe cases of Chagas disease.This was reflected in the lack of correlation between levels of autoantibodies and the left ventricular ejection fraction or the left ventricular end-diastolic diameter. there seems to be no association between the levels of antibodies and the degree of left ventricular function.8% patients and were related to symptoms. In the endemic area. there were significant associations between the level of anti-M2 antibodies and the presence of apical lesion. it is possible that the titers of circulating autoantibodies do not reflect the amount of those fixed on myocardium neurotransmitter receptors. although the presence of antibodies is associated with the presence of Chagas’ heart disease. The latter results suggest that left ventricular dysfunction appears not to be the cause of augmented serum levels of anti-M2 and anti-ß1 antibodies in Chagas disease and. specially palpitations. especially of the apex and the postero-inferior wall of the left ventricle. ECG abnormalities. Despite the apparent lack of association between left ventricular function and level of autoantibodies. None of the latter parameters were associated with the level of anti-ß1 receptor antibodies. are common in Chagas’ heart disease (29).31). with or without thrombus (30. aneurysms of the left ventricle were diagnosed in 18. both important parameters of left ventricular dysfunction. The apical aneurysm is the most peculiar finding in Chagas disease.
inflammatory. especially during exercise.ventricular function (32). receptor antibodies was greater in patients in whom the presence of chronotropic incompetence during exercise testing was detected. we found that there was an association between the levels of anti-M2 receptor antibodies and the presence of effortinduced ventricular arrhythmias. Our results showed that patients with apical aneurysm had higher levels of anti-M2 receptor antibodies than patients without the lesion. loss of parasympathetic function (12). left ventricular aneurysms are associated with the development of malignant sustained ventricular arrhythmias (33) and are independent predictors of death (34). as demonstrated by exercise testing. The occurrence of ventricular arrhythmia during exercise testing is related with increased mortality due to Chagas disease (35). which is independent of left ventricular dysfunction in Chagas disease. Here. Moreover. The pathogenetic hypothesis for the development of the left ventricular aneurysms includes mechanical. may be considered a manifestation of autonomic dysfunction (11). Chronotropic incompetence. ischemic and autonomic mechanisms. Preserved autonomic modulation is a major determinant of physiological heart rate response to exercise. but not anti-ß1. although the exact meaning of this finding remains unknown. The loss of parasympathetic tonus could then potentially facilitate the occurrence of arrhythmias. This is consistent with our previous results 12 . the level of anti-M2. Indeed. The latter findings may be secondary to chronic muscarinic receptor stimulation by the antibodies and resulting receptor desensitization and impairment of parasympathetic function. Experimental studies have shown that activation of muscarinic receptors by anti-receptor antibodies may induce their internalization and. electrical. consequently.
Nevertheless. Overall. these results point to an important role of anti-M2 receptor antibodies in the pathogenesis of the dysautonomia frequently observed in Chagas disease. the levels of anti-M2 antibodies were greater in patients with apical aneurysm. we showed that the levels of anti-M2 or anti-ß1 receptors in serum of patients with Chagas disease seemed not to correlate with the severity of the left ventricular dysfunction. 13 . in those with exercise-induced ventricular arrhythmias and were related to the presence of chronotropic incompetence. In conclusion.demonstrating the presence of parasympathetic impairment in the absence of heart failure in Chagas disease (11).
and University of Buenos Aires (UBACYT).Acknowledgements This investigation received financial support from FAPEMIG. 14 . the UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases (TDRWHO A970728 and A20771) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq-Brazil) and National Research Council (CONICET) of Argentina. Argentina. Buenos Aires.
2001. The role of endothelin in the pathogenesis of Chagas’disease. Silva. Pestell RG. Teixeira.T.M. Gazzinelli. Soares MB. Autoimmunity 2001. Douglas SA. The pathogenesis of Chagas' disease: when autoimmune and parasite-specific immune responses meet. Clin Exp Immunol 1984. Teixeira HC. 31: 499-511. inflammation and Trypanosoma cruzi infection.. Jelicks LA. Parasite persistence in the aetiology of Chagas disease. Factor SM. Arana R. Chronic Chagas’disease cardiomyopathy patients display an increased IFN-g response to Trypanosoma cruzi infection. Petkova SB. Chemokines. Trends Parasitol. 2003.. Cossio P. Tarleton RL. Kalil J. 73: 547-59. Int J Parasitol. 3. The significance of autoimmunity in the pathogenesis of Chagas heart disease. Rizzo LV. Ribeiro-Dos-Santos R. Bocchi EA.References 1. Circulatin IgG in Chagas’disease which binds to h-adrenoreceptores of myocardium and modulate their activity. Huang H. 2001. 7. de la Vega M. Pontes-De-Carvalho L. J. Bouzahzah B. R. Intern J. 17:99-108. Leon JS. 15 . Parasitol. Tanowitz HB. Wittner M.S. Borda E. Cunha-Neto E. Weiss LM. Carrara D. Front Biosci. 31:550-554. Engman DM. 2. J. Abel LCJ. 5.18: 262-5. Mady C. An Acad Bras Cienc 2001. 6. 57:670-86. 8:315-322. Ianni B. 2002. 4. Albuquerque F. Sterin-Borda L. M. Bacal F.
9. Borda E. Ribeiro AL. Garcia G. Elkayan V. Iosa D. Dequatro V. Caieiro T. 2001. 13. Ribeiro AL. Palmero H. Kaohout T. 1989. Alterations in cardiac beta-adrenergic receptors in chagasic mice and their association with circulating betaadrenoceptor-related autoantibodies. Dequatro V. Antiadrenergic and muscarinic receptor antibodies in Chagas cardiomyopathy. Gonzalez Cappa S. Cardiovas Res 1999. 127:169-177. 41:116-25. Postan M. Borda ES & Sterin-Borda L. Moraes RS.8. Intern J Cardiol 1996.. T. Pathogenesis of cardiac neuropathy in Chagas Disease and the role of autonomic nervous system. Rocha MOC. J. Oliveira E. Joensen L. 11. Parasympathetic dysautonomia precedes left ventricular systolic dysfunction in Chagas disease. Perry S. De-Ping-Lee D. Gorelik G. 2003. Sterin-Borda L. Plasma norepinephrine in chagasic cardiomyopathy: a marker of progressive dysautonomia. 30:583-588. Front Biosc. 14. Am Heart J. Torres RM.. 1990. 12.cruzi antigen that interacts with the beta 1 adrenergic receptor and modifies myocardial contractile activity. Iosa D. Rocha MO. Ferlin EL. 8:E44-54. De-Ping-Lee D. Autonomic Nervous System. Borda E. Sterin-Borda L. 10. 141:260-265. 54:149-56. Mol Biochem Parasitol 2003. Clinical Management of chronic Chagas cardiomyopathy. Teixeira MM. Am Heart J. 117: 882-887. Ribeiro JP. Elkayan V. 16 .
Pacing Clin Electrophysiol. 16. Sadoul N. Goin JC. Hernando AC. Bellotti G. Luz PL. 19:477-83. 19. 49:699-705. Reichek N. Iantorno G. 20. 2000. Cardoso RHA. Esteva M. Ann N Y Acad Sci.15. Circulation 1994. 34:1645-1654. Survival and predictors of survival in patients with congestive heart failure due to Chagas'cardiomyopathy. McKenna WJ. Schiller NB. Bilder CR. Bayo Hanza C. Schnittger I. Sahn D. Devereux R. Gutgesell H. 17 . Sterin-Borda L. J Mol Cell Cardiol 2002. Fei L.90:3098-3102. Sterin-Borda L. Therapeutic used of muscarinic acetylcholine receptor peptide to prevent mice chagasic cardiac dysfunction. 1996. Crawford M. Keeling PJ. Gut 2001. Pileggi F. Role of neurotransmitter autoantibodies in the pathogenesis of chagasic peripheral dysautonomia.2:358-367. American Society of Echocardiography Committee on Standards. Malik M. Camm AJ. Borda E. Recommendations for quantitation of the left ventricle by two-dimensional echocardiography. Sterin-Borda L. Interaction of human chagasic IgG with human colon muscarinic acetylcholine receptor: molecular and functional evidence. Feigenbaum H. Subcommittee on Quantitation of Two-Dimensional Echocardiograms. Decreased heart rate variability in patients with congestive heart failure and chronotropic incompetence. Borda E. 18. 917: 273-80. DeMaria A. J Am Soc Echocardiogr 1989. 17. Borda E. Copie X. Mady C. Joensen L. Barretto ACP. Shah PM.
Masuda MO. 6:476-83. Bergami PL. 27. Tona F. 12:175-7. Eur Heart J. Eur J Heart Fail 2002. Borda E. 24. 23. b-receptor antibodies and genetics in dilated cardiomyopathy. 272:12989-12993. 12:1551-8. Campos de Carvalho AC. 63:221-30. Goin JC. Pedrosa RC. Bestetti RB. Desensitization and sequestration of human M2 muscarinic acetylcholine receptors by autoantibodies from patients with Chagas’ disease. Rev. Dos Santos Almeida NA. de Oliveira SF. 96:2031-7. 4:411-417. Limas C. Autoimmunity against cardiovascular receptors: structural and functional implications. Perez Leiros C. Circulating cardiac autoantibodies in dilated cardiomyopathy and myocarditis: pathogenetic and clinical significance. 1998. Lima CJ. Nascimento JH. Hoebeke J. J Biol Chem 1997. 18 .21. Trop Med Int Health. Argent. Hosey M. 1995. 2001. Dos Santos Costa PC. De Oliveira SF. Cardiol. FASEB J. Masuda MO. 25. Functionally active cardiac antibodies in chronic Chagas' disease are specifically blocked by Trypanosoma cruzi antigens. Sterin-Borda L. 22. Circulation 1997. Hoebeke J. Caforio ALP. Pedrosa RC. McKenna WJ. Campos de Carvalho AC. Levin M. Mahon NJ. Predictors of unfavourable prognosis in chronic Chagas’disease. Sera from chronic chagasic patients with complex cardiac arrhythmias depress electrogenesis and conduction in isolated rabbit hearts. 1991. Ferrari I. 26.
Pedroso ERP. Silva JLL. Teixeira MM. Suarez JA. Pinheiro R. Med. Lustig L. Clinical and epidemiological aspects of Chagas disease. de Paola AA. C. Horowitz LN. 4:249-255. A lesão cardíaca vorticilar associada à doença de Chagas [The apical lesion of the heart in Chagas´disease]. 31:457-463. 65: 360-3. Prata A. 13: 179-182. 34. 19 . Rev.28. Guiguer N Jr. Borda E. Xavier SS. Rocha MOC. Sterin-Borda L. Am J Cardiol 1990. Ferreira DF. Rahimtoola SH. Perez Leiros C. IV. Brazil. 1998. Clin Auton Res 1994. 32. Pirmez. 29. Soc. 31. Cirenza C. Giordano H. N Engl J Med 1991. 33. Acquatella H. 1980. Lancet Infect Dis 2001. Rev. Chagas' disease in Virgem da Lapa County. Clinical and epidemiological aspects of the left ventricle aneurism. Minas Gerais 2003. Puigbo JJ. Portugal OP. Hirschhaut E. Trop. 62: 787-99. Casal H. Chagas' heart disease in the United States. Schiller NB. Circulation. Borges-Pereira J. 30. Arreaza N. 1: 92-100. Minas Gerais State. Bras. Goren N. Angiographic and electrophysiologic substrates of ventricular tachycardia in chronic Chagasic myocarditis. Hagar JM. 325: 763-768. Terzian AB. A clinical and pathologic study. Valecillos R. M-mode and two-dimensional echocardiography in chronic Chages' heart disease. Alterations in cardiac muscarinic acetylcholine receptors in mice with autoimmune myocarditis and assiociation with circulating muscarinic receptor-related autoantibodies. Miyamoto MH. Med.
1995. 74: 293-5. Terzian AB. Gomes JA. Ventricular tachycardia during exercise testing as a predictor of sudden death in patients with chronic chagasic cardiomyopathy and ventricular arrhythmias. 20 . de Paola AA. Br Heart J. Martinez Fo EE.35. Miyamoto MH.
Legends of Figures Figure 1. 0. Figure 2. Dots 21 .200. Figure 3.100. Dots represent the individual optical density (OD) values for each serum sample at 1/50 dilution from infected patients in the absence (30 patients) or in the presence (22 patients) of left ventricular apical lesions. Scattergram showing immunoreactivity of serum anti-M2 cholinergic (A) and anti-β1 adrenergic (B) autoantibodies tested by ELISA. and anti-β1 adrenergic. Dotted line shows cutoff values (mean OD values ± 2 SD from NI group): anti-M2 cholinergic. Horizontal lines show median OD values.0005 versus NI. Chagasic patients were submitted to a treadmill exercise and the presence or absence of (A) exercise-dependent ventricular arrhythmia or (B) chronotropic defect evaluated. 0. Distribution of anti-M2 cholinergic and anti-β1 adrenergic cholinergic autoantibodies in sera of chagasic patients with apical lesion. *p < 0. Dots represent the individual optical density (OD) values for each serum sample at 1/50 dilution from 6 non-infected (NI). 8 infected patients in indeterminate form (IND) and 52 chronic chagasic cardiomyopathy (CCC). Distribution of anti-M2 cholinergic and anti-β1 adrenergic autoantibodies in sera of chagasic patients with extra systolic beats and chronotropic incompetence. Horizontal lines show median OD values.
represent the individual optical density (OD) values for each serum sample at 1/50 dilution. 22 . Horizontal lines show median OD values.
Non-normally distributed data were transformed before performing ANOVA and means comparisons. IND: indeterminate form. except for NVPB in 24h that are shown as median [25%-75% percentile].Table 1. 23 .9 47 ± 3.0 43 ± 2. NVPB: number of ventricular premature beats in 24 h. NI: non-infected. LVEF: left ventricle ejection fraction. # P < 0. ________________________________________________________________________ NI (n=6) IND (n=8) I (n=8) II/III (n=7) IV (n=15) V (n=14) Age 38 ± 6.5 44 ± 2.3 Gender (% male) 67 38 50 29 47 79 LVEF (%) 68 ± 6 64 ± 4 66 ± 2 60 ± 3 61 ± 2 45 ± 3# LVDD (mm) NVPB in 24 h 48 ± 1 48 ± 1 49 ± 1 47 ± 2 50 ± 1 62 ± 1# ND 1 [0-5] 3 [0-258] 725 [399-5676] 86 [18-2863] 840 [192-3002] Values are shown as mean ± SD.4 48 ± 8.2 47 ± 4.01 when compared to non-infected and furthermore chagasic individuals. LVDD: left ventricle diastolic diameter. Clinical characteristics of non-infected individuals (NI) and patients classified with different levels of chronic chagasic cardiomyopathy (CCC).
100. 24 .7 0/6 6/8 5/8 4/7 13/15 9/14 0 77. Distribution of anti-M2 cholinergic and anti-β1 adrenergic autoantibodies in non-infectd individuals and infected patients anti-M2 cholinergic Groups number positive/total percentage anti-β1 adrenergic number positive/total percentage NI IND CCC I CCC II/III CCC IV CCC V 0/6 3/8 7/8 5/7 14/15 12/14 0 37.3 85.1 86.Table 2.5* 87.3 ______________________________________________________________________________ Microtitre wells were coacted with 1 µg peptides (anti-M2 and anti-β1) and ELISA was carried out in the presence of sera from non-infected individuals (NI).1 62. Prevalence values of anti-M2 cholinergic and anti-β1 adrenergic autoantibodies differ with *p < 0. indeterminate form (IND) and different degree of chronic chagasic cardiomyopathy (CCC I to V) infected patients.5 71. Cutoff values for anti-M2 cholinergic 0.4 93. Optical density (OD) values more than 2 SD about normal mean were taken as positive.5 57.6 64.0005 versus CCC I to V.200 and for anti-β1 adrenergic 0.
973 R= 0.406 R= 0. NVPB: number of ventricular premature beats.858 R= 0.Table 3.725 R=0.052 p= 0.027 p= 0.053 p= 0. LVEF (%) LVEDD (mm) NVPB (in 24 h) Anti-adrenergic antibodies Anti-cholinergic antibodies R= 0. Lack of correlation between parameters of ventricular dysfunction and levels of anti-adrenergic or anti-cholinergic receptor antibodies in patients with chronic chagasic cardiomypathy (CCC). 25 .167 p= 0.120 p= 0. LVEDD: left ventricle end-diastolic diameter.270 LVEF: left ventricle ejection fraction.005 p= 0.733 R= 0.
50 0.0 NI All forms IND I II/III IV V Chronic Chagasic cardiomyopathy B 0.25 0.0 0.5 0.Figure 1 A 1.75 anti-ß1 OD (405 nm) 0.5 anti-M2 OD (405 nm) 1.00 NI All forms IND I II/III IV V Chronic Chagasic cardiomyopathy 26 .
Figure 2: p<0.001 1.0 0.0 Absence Presence Absence Presence anti-M2 anti-ß1 Left ventricle apical lesion 27 .5 0.5 OD (405 nm) 1.
5 0.001 1.Figure 3: A p<0.0 Absence Presence Absence Presence anti-M1 anti-ß2 Chronotropic incompetence 28 .5 0.0 Absence Presence Absence Presence anti-M1 anti-ß2 Exercise-induced ventricular arrthymia B p<0.5 OD (405 nm) 1.5 OD (405 nm) 1.0 0.0 0.05 1.
This action might not be possible to undo. Are you sure you want to continue?
We've moved you to where you read on your other device.
Get the full title to continue listening from where you left off, or restart the preview.