Optimized viral dose and transient immunosuppression enable herpes simplex virus ICP0-null mutants to establish wild-type levels of latency in vivo

被引:44
作者
Halford, WP [1 ]
Schaffer, PA [1 ]
机构
[1] Univ Penn, Sch Med, Dept Microbiol, Philadelphia, PA 19104 USA
关键词
D O I
10.1128/JVI.74.13.5957-5967.2000
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The reduced efficiency with which herpes simplex virus type 1 (HSV-1) mutants establish latent infections in vivo has been a fundamental obstacle in efforts to determine the roles of individual viral genes in HSV-1 reactivation. For example, in the absence of the "nonessential" viral immediate-early protein, ICP0, HSV;1 is severely impaired in its ability to (i) replicate at the site of inoculation and (ii) establish latency in neurons of the peripheral nervous system, The mouse ocular model of HSV latency was used in the present study to determine if the conditions of infection can be manipulated such that replication-impaired, ICP0-null mutants establish wild-type levels of latency, as measured by viral genome loads in latently infected trigeminal ganglia (TG), To this end, the effects of inoculum size and transient immunosuppression on the levels of acute replication in mouse eyes and of viral DNA in latently infected TG were examined. Following inoculation of mice with 2 x 10(3), 2 x 10(4), 2 x 10(5), or 2 x 10(6) PFU/eye, wild-type virus replicated in mouse eyes and established latency in TG with similar efficiencies at all four doses. In contrast, increasing the inoculum size of the ICP0-null mutants n212 and 7134 from 2 x 10(5) to 2 x 10(6) PFU/eye significantly decreased the levels of infectious virus detected in the tear films of mice from days 4 to 9 postinfection. In an attempt to establish the biological basis for this finding, the effect of viral dose on the induction of the host proinflammatory response was examined. Quantitative reverse transcription-PCR demonstrated that increasing the inoculum of 7134 from 2 x 10(5) to 2 x 106 PFU/eye significantly increased the expression of proinflammatory (interleukin 6), cell adhesion (intercellular adhesion molecule 1), and phagocyte-associated (CD11b) genes in mouse eyes 24 h postinfection. Furthermore, transient immunosuppression of mice with cyclophosphamide, but not cyclosporin A, significantly enhanced both the levels of acute n212 and 7134 replication in the eye and the levels of mutant viral genomes present in latently infected TG in a dose;dependent manner, Thus, the results of this study demonstrate that acute replication in the eye and the number of ICP0-null mutant genomes in latently infected TG can be increased to wild-type levels for both n212 and 7134 by (i) optimization of inoculum size and (ii) transient immunosuppression with cyclophosphamide.
引用
收藏
页码:5957 / 5967
页数:11
相关论文
共 61 条
  • [21] Immunization with HSV-1 antigen rapidly protects against HSV-1-induced encephalitis and is IFN-γ independent
    Halford, WP
    Veress, LA
    Gebhardt, BM
    Carr, DJJ
    [J]. JOURNAL OF INTERFERON AND CYTOKINE RESEARCH, 1998, 18 (03) : 151 - 158
  • [22] HERPES-SIMPLEX VIRUS TYPE-1 IMMEDIATE-EARLY PROTEIN VMW110 REACTIVATES LATENT HERPES-SIMPLEX VIRUS TYPE-2 IN AN INVITRO LATENCY SYSTEM
    HARRIS, RA
    EVERETT, RD
    ZHU, XX
    SILVERSTEIN, S
    PRESTON, CM
    [J]. JOURNAL OF VIROLOGY, 1989, 63 (08) : 3513 - 3515
  • [23] Quantitative analysis of polymerase chain reaction products by dot blot
    Hill, JM
    Halford, WP
    Wen, RJ
    Engel, LS
    Green, LC
    Gebhardt, BM
    [J]. ANALYTICAL BIOCHEMISTRY, 1996, 235 (01) : 44 - 48
  • [24] HERPES-SIMPLEX VIRUS LATENT PHASE TRANSCRIPTION FACILITATES INVIVO REACTIVATION
    HILL, JM
    SEDARATI, F
    JAVIER, RT
    WAGNER, EK
    STEVENS, JG
    [J]. VIROLOGY, 1990, 174 (01) : 117 - 125
  • [25] QUIESCENT VIRAL GENOMES IN HUMAN FIBROBLASTS AFTER INFECTION WITH HERPES-SIMPLEX VIRUS TYPE-1 VMW65 MUTANTS
    JAMIESON, DRS
    ROBINSON, LH
    DAKSIS, JI
    NICHOLL, MJ
    PRESTON, CM
    [J]. JOURNAL OF GENERAL VIROLOGY, 1995, 76 : 1417 - 1431
  • [26] Jordan Robert, 1997, Journal of Virology, V71, P6850
  • [27] QUANTITATIVE POLYMERASE CHAIN-REACTION ANALYSIS OF HERPES-SIMPLEX VIRUS-DNA IN GANGLIA OF MICE INFECTED WITH REPLICATION-INCOMPETENT MUTANTS
    KATZ, JP
    BODIN, ET
    COEN, DM
    [J]. JOURNAL OF VIROLOGY, 1990, 64 (09) : 4288 - 4295
  • [28] Interferons regulate the phenotype of wild-type and mutant herpes simplex viruses in vivo
    Leib, DA
    Harrison, TE
    Laslo, KM
    Machalek, MA
    Moorman, NJ
    Virgin, HW
    [J]. JOURNAL OF EXPERIMENTAL MEDICINE, 1999, 189 (04) : 663 - 672
  • [29] A DELETION MUTANT OF THE LATENCY-ASSOCIATED TRANSCRIPT OF HERPES-SIMPLEX VIRUS TYPE-1 REACTIVATES FROM THE LATENT STATE WITH REDUCED FREQUENCY
    LEIB, DA
    BOGARD, CL
    KOSZVNENCHAK, M
    HICKS, KA
    COEN, DM
    KNIPE, DM
    SCHAFFER, PA
    [J]. JOURNAL OF VIROLOGY, 1989, 63 (07) : 2893 - 2900
  • [30] IMMEDIATE-EARLY REGULATORY GENE MUTANTS DEFINE DIFFERENT STAGES IN THE ESTABLISHMENT AND REACTIVATION OF HERPES-SIMPLEX VIRUS LATENCY
    LEIB, DA
    COEN, DM
    BOGARD, CL
    HICKS, KA
    YAGER, DR
    KNIPE, DM
    TYLER, KL
    SCHAFFER, PA
    [J]. JOURNAL OF VIROLOGY, 1989, 63 (02) : 759 - 768