Screening of an Echinococcus granulosus cDNA library with IgG4 from patients with cystic echinococcosis identifies a new tegumental protein involved in the immune escape.
The worldwide problem of chronic Echinococcus granulosus disease calls for new parasite-derived immunomodulatory molecules. By screening an E. granulosus cDNA library with IgG4 from patients with active cystic echinococcosis, we identified a cDNA that encodes a predicted partial protein that immunofluorescence studies localized in the protoscolex tegument and on the germinal layer of cyst wall. We named this protein EgTeg because the 105 amino acid sequence scored highest against a family of Schistosoma tegumental proteins. Evaluating the role of EgTeg in the human early inflammatory response we found that EgTeg significantly inhibited polymorphonuclear cell (PMN) chemotaxis. Cytometric analysis of intracellular cytokines disclosed a significantly higher percentage of cells producing IL-4 than IFN-gamma (P = 0.001, Student's t-test) in T lymphocytes from patients with cystic echinococcosis stimulated with EgTeg. EgTeg induced weak Th1-dependent proliferation in 42% of patients' peripheral blood mononuclear cells. In immunoblotting (IB) analysis of total IgG and IgG subclass responses to EgTeg in patients with cystic echinococcosis, patients with other parasitoses, patients with cystic lesions and healthy controls, total IgG specific to EgTeg yielded high sensitivity (73%) but low specificity (44%) precluding its use in immunodiagnosis. Conversely, IgG4 specific to EgTeg gave acceptable sensitivity (65%) and high specificity (89%) suggesting its use in immunodiagnosis to confirm ultrasound documented cysts suggestive of E. granulosus. Because the new tegumental antigen EgTeg inhibits chemotaxis, induces IL-4-positive T lymphocytes and noncomplement fixing antibodies (IgG4) it is an immunomodulatory molecule associated with chronic infection.
Ortona
E
E
Division of Immune-mediated Diseases, Department of Infectious, Parasitic and Immune-mediated Diseases, Istituto Superiore di Sanità, Rome, Italy.
Margutti
P
P
Delunardo
F
F
Nobili
V
V
Profumo
E
E
Riganò
R
R
Buttari
B
B
Carulli
G
G
Azzarà
A
A
Teggi
A
A
Bruschi
F
F
Siracusano
A
A
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000032:analysis / Q000276:immunology*; D001483:Base Sequence; D002465:Cell Movement / Q000276:immunology; D002634:Chemotaxis, Leukocyte / Q000276:immunology; D003560:Cysts / Q000276:immunology; D004269:DNA, Bacterial / Q000276:immunology; D004270:DNA, Circular / Q000276:immunology; D004443:Echinococcosis / Q000276:immunology*; D048209:Echinococcus granulosus / Q000276:immunology*; D015723:Gene Library; D006801:Humans; D007111:Immunity, Cellular / Q000276:immunology; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000276:immunology*; D007150:Immunohistochemistry / Q000379:methods; D007371:Interferon-gamma / Q000032:analysis / Q000276:immunology; D015847:Interleukin-4 / Q000032:analysis / Q000276:immunology; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000276:immunology; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004270:DNA, Circular; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D015847:Interleukin-4; D007371:Interferon-gamma