The heavy chain variable segment gene repertoire in chronic Chagas' heart disease.
Patients chronically infected with Trypanosoma cruzi develop chronic Chagas' heart disease (cChHD). Their Ab response is suspected to be involved in the cardiac pathogenesis. Reactivity of serum Abs from these patients has been extensively studied but little is known about the diversity of the in vivo IgG repertoire. We analyzed 125 variable H chain (VH) genes and compared it to repertoires from healthy individuals, and patients with autoimmune processes and other infections. VH were from plasma cells isolated from heart tissue of three cChHD patients and from a Fab combinatorial library derived from bone marrow of another cChHD patient. The role of the parasite in shaping the Ab repertoire was assessed analyzing VH genes before and after panning against T. cruzi Ag. Among recovered VH genes, a significantly increased representation of VH4 was observed. Plasma cells at the site of cardiac infiltration showed an increased VH1 usage. CDR3 lengths were similar to the ones found in the healthy repertoire and significantly shorter than in other infections. VH derived from anti-T. cruzi Fab and plasma cells showed a higher proportion of hypermutated genes, 46.9% and 43.75%, respectively, vs 30.9% of the cChHD patient repertoire, pointing to the role of parasite Ags in the shaping of the humoral response in Chagas' disease. No histological evidence of germinal center-like structures was observed in heart tissue. In accordance, VH analysis of heart plasmocytes revealed no evidence of clonal B cell expansion, suggesting that they migrated into heart tissue from secondary lymphoid organs.
Grippo
Vanina
V
Laboratory of Molecular Biology of Chagas' Disease, Institute for Genetic Engineering and Molecular Biology, CONICET, Buenos Aires, Argentina.
Mahler
Evelyn
E
Elias
Fernando E
FE
Cauerhff
Ana
A
Gómez
Karina A
KA
Tentori
Maria C
MC
Ruiz
Aurora
A
Vigliano
Carlos A
CA
Laguens
Ruben P
RP
Berek
Claudia
C
Levin
Mariano J
MJ
D000328:Adult; D000595:Amino Acid Sequence; D000913:Antibodies, Protozoan / Q000096:biosynthesis / Q000235:genetics*; D000953:Antigens, Protozoan / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology / Q000469:parasitology / Q000473:pathology; D002598:Chagas Cardiomyopathy / Q000276:immunology* / Q000469:parasitology / Q000473:pathology; D002908:Chronic Disease; D022801:Complementarity Determining Regions / Q000096:biosynthesis / Q000235:genetics; D005260:Female; D015322:Gene Rearrangement, B-Lymphocyte / Q000235:genetics*; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis* / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis* / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D027041:Somatic Hypermutation, Immunoglobulin / Q000235:genetics; D014349:Trypanosoma cruzi / Q000276:immunology
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D022801:Complementarity Determining Regions; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region