Subject(s)
Chagas Disease/blood , Chagas Disease/diagnosis , Chemokine CXCL10/blood , Proto-Oncogene Proteins c-sis/blood , Aged , Becaplermin , Biomarkers/blood , Chronic Disease , Female , Humans , Male , Middle AgedABSTRACT
BACKGROUND: L-ficolin (encoded by FCN2) binds to acetylated sugar moieties of many pathogens, including Trypanosoma cruzi, promoting their phagocytosis and lysis by the complement system. METHODS: We investigated L-ficolin levels in 160 T. cruzi infected patients with chronic Chagas disease and 71 healthy individuals, and FCN2 polymorphisms (-986 G>A, -602 G>A, and -4 A>G in the promoter and A258S in exon 8) in 243 patients, being 88 indeterminate (asymptomatic), 96 with cardiac, 23 with digestive and 33 with cardiodigestive manifestations (two were unspecified) and 305 controls (135 for A258S). RESULTS: Patients presented lower L-ficolin plasma levels than controls (p<0.0001). Among the different groups of cardiac commitment, individuals with moderate forms had higher L-ficolin levels than the severe forms (Pâ=â0.039). Lower L-ficolin levels were found associated with the 258S variant in the patients (Pâ=â0.034). We found less -4A/G heterozygotes in the cardiac patients, than in the controls (ORâ=â0.56 [95% CIâ=â0.33-0.94], Pâ=â0.034). Heterozygote -4A/G genotypes with the 258S variant and 258SS homozygotes were nevertheless more frequent among cardiodigestive patients than in controls (ORâ=â14.1 [95% CIâ=â3.5-56.8], Pâ=â0.0001) and in indeterminate patients (ORâ=â3.2 [95% CIâ=â1.1-9.4], Pâ=â0.037). We also found an association of the allelic frequency of the 258S variant with cardiodigestive Chagas disease compared to controls (ORâ=â2.24 [95% CIâ=â1.1-4.5], Pâ=â0.037). Thus, decreased patient levels of L-ficolin reflect not only protein consumption due to the disease process, but also the higher frequency of the 258S variant in patients with cardiodigestive symptoms. CONCLUSION: The very first study on Brazilian cohort associates both L-ficolin plasma levels and FCN2 variants to Chagas disease and subsequent disease progression. The prognostic value of L-ficolin levels and the FCN2*A258S polymorphism should be further evaluated in other settings.
Subject(s)
Chagas Disease/blood , Chagas Disease/genetics , Genotype , Lectins/blood , Lectins/genetics , Adult , Aged , Aged, 80 and over , Alleles , Brazil , Case-Control Studies , Chronic Disease , Female , Gene Frequency , Genetic Predisposition to Disease , Haplotypes , Humans , Male , Middle Aged , Polymorphism, Single Nucleotide , FicolinsABSTRACT
BACKGROUND: Ficolin-2 coded by FCN2 gene is a soluble serum protein and an innate immune recognition element of the complement system. FCN2 gene polymorphisms reveal distinct geographical patterns and are documented to alter serum ficolin levels and modulate disease susceptibility. METHODS: We employed a real-time PCR based on Fluorescence Resonance Energy Transfer (FRET) method to genotype four functional SNPs including -986 G > A (#rs3124952), -602 G > A (#rs3124953), -4A > G (#rs17514136) and +6424 G > T (#rs7851696) in the ficolin-2 (FCN2) gene. We characterized the FCN2 variants in individuals representing Brazilian (n = 176), Nigerian (n = 180), Vietnamese (n = 172) and European Caucasian ethnicity (n = 165). RESULTS: We observed that the genotype distribution of three functional SNP variants (-986 G > A, -602 G > A and -4A > G) differ significantly between the populations investigated (p < 0.0001). The SNP variants were highly linked to each other and revealed significant population patterns. Also the distribution of haplotypes revealed distinct geographical patterns (p < 0.0001). CONCLUSIONS: The observed distribution of the FCN2 functional SNP variants may likely contribute to altered serum ficolin levels and this may depend on the different disease settings in world populations. To conclude, the use of FRET based real-time PCR especially for FCN2 gene will benefit a larger scientific community who extensively depend on rapid, reliable method for FCN2 genotyping.
Subject(s)
Genetic Variation , Lectins/genetics , Asian People/genetics , Black People/genetics , Fluorescence Resonance Energy Transfer/methods , Haplotypes/genetics , Humans , Lectins/blood , Polymorphism, Single Nucleotide , Real-Time Polymerase Chain Reaction , Sequence Analysis, DNA/methods , White People/genetics , FicolinsABSTRACT
Mannose-binding lectin (MBL) initiates complement on Trypanosoma cruzi through the MBL-associated serine protease 2 (MASP2). We haplotyped six MASP2 polymorphisms in 208 chronic chagasic patients, being 81 indeterminate and 123 symptomatic (76 with cardiac, 19 with digestive and 28 with cardiodigestive forms) and 300 healthy individuals from Southern Brazil, using PCR with sequence-specific primers. The g.1961795C, p.371D diplotype (short CD) occurred at a higher frequency among symptomatic patients, compared with the indeterminate group (P(Bf)=0.012, OR=3.11), as well as genotypes with CD, but not with the g.1945560A in the promoter in cardiac patients (P(Bf)=0.012, OR=13.54). CD haplotypes linked to the p.P126L and p.V377A variants were associated with reduced MASP-2 levels (P<0.0001) but not reduced MBL/MASP-2/C4 complexes. MASP2*CD genotypes, most of them generating low MASP-2 levels, are associated with high risk of chagasic cardiomyopathy. Rapid MASP2 genotyping might be used to predict the risk of symptomatic disease.
