scholarly journals A nonstructural polypeptide encoded by immediate-early transcription unit 1 of murine cytomegalovirus is recognized by cytolytic T lymphocytes.

1987 ◽  
Vol 166 (1) ◽  
pp. 289-294 ◽  
Author(s):  
U H Koszinowski ◽  
G M Keil ◽  
H Schwarz ◽  
J Schickedanz ◽  
M J Reddehase
Keyword(s):  
Regulatory Protein ◽  
Antigen Expression ◽  
Transcription Unit ◽  
Immediate Early ◽  
Murine Cytomegalovirus ◽  
Target Cells ◽  
Cytolytic T Lymphocytes ◽  
Gene Encoding ◽  
Dna Coding ◽  
Early Transcription

We have constructed target cells by cotransfection of the MHC gene Ld and fragments of murine cytomegalovirus (MCMV) DNA coding for nonstructural immediate-early (IE) proteins. Transfectants were tested by using CTL clone IE1 with specificity for an IE epitope presented in association with Ld. Data show that clone IE1 recognizes a product of the ie1 transcription unit of MCMV, and that its specificity is shared by approximately 25% of polyclonal IE-specific CTL. The results provide the first definite evidence that expression of a herpes virus IE gene encoding a regulatory protein gives rise to antigen expression detectable by specific CTL.

scholarly journals Cytolytic T lymphocyte recognition of the murine cytomegalovirus nonstructural immediate-early protein pp89 expressed by recombinant vaccinia virus.

1987 ◽  
Vol 166 (3) ◽  
pp. 668-677 ◽  
Author(s):  
H Volkmer ◽  
C Bertholet ◽  
S Jonjić ◽  
R Wittek ◽  
U H Koszinowski
Keyword(s):  
Vaccinia Virus ◽  
Antigen Expression ◽  
Recombinant Vaccinia Virus ◽  
Virus Genome ◽  
Site Directed Mutagenesis ◽  
Immediate Early ◽  
Murine Cytomegalovirus ◽  
Early Protein ◽  
Infected Cells ◽  
Essential Function

The murine immediate-early (IE) protein pp89 is a nonstructural virus-encoded phosphoprotein residing in the nucleus of infected cells, where it acts as transcriptional activator. Frequency analysis has shown that in BALB/c mice the majority of virus-specific CTL recognize IE antigens. The present study was performed to assess whether pp89 causes membrane antigen expression detected by IE-specific CTL. Site-directed mutagenesis has been used to delete the introns from gene ieI, encoding pp89, for subsequent integration of the continuous coding sequence into the vaccinia virus genome. After infection with the vaccinia recombinant, the authentic pp89 was expressed in cells that became susceptible to lysis by an IE-specific CTL clone. Priming of mice with the vaccinia recombinant sensitized polyclonal CTL that recognized MCMV-infected cells and transfected cells expressing pp89. Thus, a herpesviral IE polypeptide with essential function in viral transcriptional regulation can also serve as a dominant antigen for the specific CTL response of the host.

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