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T‐cell clones derived by CD3 stimulation from hepatitis C virus explanted liver tissue are not representative of dominant clones present in vivo
Author(s) -
Davey Michael P.,
Rosen Hugo R.,
Woody Carolyn N.,
Haley Daniel P.,
Kurkinen Julie,
Lewinsohn David M.
Publication year - 2001
Publication title -
liver transplantation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.814
H-Index - 150
eISSN - 1527-6473
pISSN - 1527-6465
DOI - 10.1053/jlts.2001.26058
Subject(s) - clone (java method) , t cell receptor , biology , t cell , virology , cd3 , hepatitis c virus , in vivo , antigen , in vitro , cloning (programming) , virus , immunology , gene , microbiology and biotechnology , cd8 , genetics , immune system , computer science , programming language
Liver tissue from hepatitis C virus (HCV)‐related end‐stage disease contains T‐cell infiltrates. The goal of this study is to determine whether CD4 T‐cell clones established in vitro using an antigen‐independent technique from explanted liver tissue (n = 3) are representative of dominant clones present in vivo. T‐cell receptor (TCR) use by intrahepatic CD4 T cells was assessed by spectratype analysis. Clones were established from single CD4 T cells by culturing in vitro with anti‐CD3 and interleukin‐2 (n > 25 per patient). TCR genes expressed by each clone were identified by sequencing. When identical clones were isolated, the original spectratype was analyzed further to determine whether the clone was a dominant T‐cell expansion in vivo. Evidence for clonal expansions was found in all patients by spectratyping. T cells expressing the same TCRBV genes used for spectratyping were cloned in vitro. Duplicate clones expressing the same TCR genes were observed in 2 patients. Confirmation that clones established in vitro matched those present in vivo was obtained for 2 clones. Many dominant clones identified by spectratyping did not proliferate in vitro. Although spectratyping suggested the widespread accumulation of clonal expansions in HCV‐related end‐stage liver disease, clones established in vitro using anti‐CD3 were poorly representative of dominant clones present in vivo. Although cloning with anti‐CD3 has the advantage of generating T‐cell clones not biased a priori toward a specific antigen, modified cloning strategies will need to be developed to expand those clones that appear dominant in end‐stage organs.