In vitro EBV-infected subline of KMH2, derived from Hodgkin lymphoma, expresses only EBNA-1, while CD40 ligand and IL-4 induce LMP-1 but not EBNA-2.
In about 50% of classical Hodgkin lymphomas, the Hodgkin/Reed Sternberg (H/RS) cells carry Epstein-Barr virus (EBV). The viral gene expression in these cells is restricted to EBNA-1, EBERs, LMP-1 and LMP-2 (type II latency). The origin of H/RS cells was defined as crippled germinal center B cells that escaped apoptosis. In spite of numerous attempts, only few typical Hodgkin lymphoma (HL) lines have been established. This suggests that the cells require survival factors that they receive in the in vivo microenvironment. If EBV is expected to drive the cells for growth in culture, the absence of EBNA-2 may explain the incapacity of H/RS cells for in vitro proliferation. In EBV carrying B lymphocytes, functional EBNA-2 and LMP-1 proteins are required for in vitro growth. For analysis of the interaction between EBV and the H/RS cells, we infected the CD21-positive HL line KMH2 with the B958 and Akata viral strains. Only EBNA-1 expression was detected in a few cells in spite of the fact that all cells could be infected. Using a neomycin-resistance-tagged recombinant EBV strain (Akata-Neo) we established an EBV-positive subline that was carried on selective medium. In contrast to the type II EBV expression pattern of H/RS cells in vivo, the KMH2 EBV cells did not express LMP-1. The EBV expression pattern could be modified in this type I subline. LMP-1 could be induced by the histone deacetylase inhibitors TSA and n-butyrate, by 5-AzaC, a demethylating agent, and by phorbol ester. None of these treatments induced EBNA-2. Importantly, exposure to CD40 ligand and IL-4 induced LMP-1 without EBNA-2 expression and lytic replication. The KMH2 EBV cells expressed LMP-2A, but not LMP-2B mRNAs. This result is highly relevant for the type II expression pattern of H/RS cells in vivo, since these stimuli can be provided by the surrounding activated T lymphocytes.
Kis
Loránd L
LL
Microbiology and Tumor Biology Center, Karolinska Institutet, Stockholm, Sweden. kis.lorand.levente@mtc.ki.se
Nishikawa
Jun
J
Takahara
Miki
M
Nagy
Noémi
N
Matskova
Liudmila
L
Takada
Kenzo
K
Elmberger
P Göran
PG
Ohlsson
Ann
A
Klein
George
G
Klein
Eva
E
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D023201:CD40 Ligand / Q000494:pharmacology*; D045744:Cell Line, Tumor; D019309:Epstein-Barr Virus Nuclear Antigens / Q000187:drug effects / Q000235:genetics* / Q000276:immunology / Q000494:pharmacology*; D004854:Herpesvirus 4, Human / Q000276:immunology*; D006689:Hodgkin Disease / Q000235:genetics / Q000276:immunology* / Q000821:virology; D006801:Humans; D015847:Interleukin-4 / Q000494:pharmacology*; D014764:Viral Proteins
D001425:Bacterial Outer Membrane Proteins; C402641:EBNA-2 protein, Human herpesvirus 4; D019309:Epstein-Barr Virus Nuclear Antigens; C091200:Lmp1 antigen, Mycoplasma; D014764:Viral Proteins; D023201:CD40 Ligand; D015847:Interleukin-4; C417029:EBV-encoded nuclear antigen 1