Spread and Replication of and Immune Response to γ134.5-Negative Herpes Simplex Virus Type 1 Vectors in BALB/c Mice

ABSTRACT We have previously shown that intracranial infection of herpes simplex virus type 1 (HSV-1) vector R8306 expressing interleukin-4 (IL-4) can abolish symptoms of experimental autoimmune encephalomyelitis, which is used as a model for human multiple sclerosis (Broberg et al., Gene Ther. 8:769-777, 2001). The aim of the current study was to search for means other than intracranial injection to deliver HSV-derived vectors to the central nervous system of mice. We also aimed to study the replication efficiency of these vectors in nervous system tissues and to elucidate the effects of the viruses on the immune response. We studied the spread and replication of the following viruses with deletions in neurovirulence gene γ134.5: R3616, R849 (lacZ transgene), R3659 (alpha-tk), R8306 (murine IL-4 transgene), and R8308 (murine IL-10 transgene). The samples were taken from trigeminal ganglia and brains of BALB/c mice after corneal, intralabial, and intranasal infection, and the viral load was examined by viral culture, HSV DNA PCR, and VP16 reverse transcription (RT)-PCR. The results show that (i) intranasal infection was the most efficient means of spread to the central nervous system (CNS) besides intracranial injection; (ii) the viruses did not grow in the culture from the brain samples, but the viral DNA persisted even until day 21 postinfection; (iii) viral replication, as observed by VP16 mRNA RT-PCR, occurred mainly on days 4 and 7 postinfection in trigeminal ganglia and to a low extent in brain; (iv) R3659, R8306, and R8308 showed reactivation from the trigeminal ganglia in explant cultures; (v) in the brain, the vectors spread to the midbrain more efficiently than to other brain areas; and (vi) the deletions in the R3659 genome significantly limited the ability of this virus to replicate in the nervous system. The immunological studies show that (i) the only recombinant to induce IL-4 mRNA expression in the brain was R8306, the gamma interferon response was very low in the brain for R3659 and R8306, and the IL-23p19 response to R8306 decreased by day 21 postinfection, unlike for the other viruses; (ii) Δγ134.5 HSV vectors modulated the subsets of the splenocytes differently depending on the transgene; (iii) R3659 infection of the nervous system induces expression and production of cytokines from the stimulated splenocytes; and (iv) HSV vectors expressing IL-4 or IL-10 induce expression and production of both of the Th2-type cytokines from splenocytes. We conclude that the intranasal route of infection is a possible means of delivery of Δγ134.5 HSV vectors to the CNS in addition to intracranial infection, although replication in the CNS remains minimal. The DNA of the HSV vectors is able to reside in the brain for at least 3 weeks. The features of the immune response to the vectors must be considered and may be exploited in gene therapy experiments with these vectors.

[1]  J. Erälinna,et al.  Immunomodulation by roquinimex decreases the expression of IL‐23 (p19) mRNA in the brains of herpes simplex virus type 1 infected BALB/c mice , 2004, Clinical and experimental immunology.

[2]  A. Salmi,et al.  IL-4 is the key regulator in herpes simplex virus-based gene therapy of BALB/c experimental autoimmune encephalomyelitis , 2004, Neuroscience Letters.

[3]  G. Cheng,et al.  Dephosphorylation of eIF-2α Mediated by the γ134.5 Protein of Herpes Simplex Virus Type 1 Is Required for Viral Response to Interferon but Is Not Sufficient for Efficient Viral Replication , 2003, Journal of Virology.

[4]  A. Salmi,et al.  Low copy number detection of herpes simplex virus type 1 mRNA and mouse Th1 type cytokine mRNAs by Light Cycler quantitative real-time PCR. , 2003, Journal of virological methods.

[5]  J. Erälinna,et al.  Semliki Forest virus infection is enhanced in Th1-prone SJL mice but not in Th2-prone BALB/c mice during Linomide-induced immunomodulation , 2002, Journal of Neuroimmunology.

[6]  A. Nesburn,et al.  Infection of BALB/c mice with a herpes simplex virus type 1 recombinant virus expressing IFN-gamma driven by the LAT promoter. , 2002, Virology.

[7]  B. Roizman,et al.  Cell Surface Major Histocompatibility Complex Class II Proteins Are Regulated by the Products of the γ134.5 and UL41 Genes of Herpes Simplex Virus 1 , 2002, Journal of Virology.

[8]  J. Erälinna,et al.  Herpes simplex virus type 1 infection induces upregulation of interleukin-23 (p19) mRNA expression in trigeminal ganglia of BALB/c mice. , 2002, Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research.

[9]  D. Knipe,et al.  Herpes Simplex Virus Vectors Elicit Durable Immune Responses in the Presence of Preexisting Host Immunity , 2002, Journal of Virology.

[10]  S. Glagov,et al.  Prevention of restenosis by a herpes simplex virus mutant capable of controlled long-term expression in vascular tissue in vivo , 2001, Gene Therapy.

[11]  G. Cheng,et al.  Val193 and Phe195 of the γ134.5 Protein of Herpes Simplex Virus 1 Are Required for Viral Resistance to Interferon-α/β , 2001 .

[12]  S. Dunnett,et al.  Latency associated promoter transgene expression in the central nervous system after stereotaxic delivery of replication-defective HSV-1-based vectors , 2001, Gene Therapy.

[13]  J. Erälinna,et al.  Expression of interleukin-4 but not of interleukin-10 from a replicative herpes simplex virus type 1 viral vector precludes experimental allergic encephalomyelitis , 2001, Gene Therapy.

[14]  K. Delman,et al.  Effects of preexisting immunity on the response to herpes simplex-based oncolytic viral therapy. , 2000, Human gene therapy.

[15]  C. Miller,et al.  Role of the immune response during neuro-attenuated herpes simplex virus-mediated tumor destruction in a murine intracranial melanoma model. , 2000, Cancer research.

[16]  K. Molnar-Kimber,et al.  Effect of preexisting anti-herpes immunity on the efficacy of herpes simplex viral therapy in a murine intraperitoneal tumor model. , 2000, Molecular therapy : the journal of the American Society of Gene Therapy.

[17]  V. Hukkanen,et al.  Time-Resolved Fluorometry PCR Assay for Rapid Detection of Herpes Simplex Virus in Cerebrospinal Fluid , 2000, Journal of Clinical Microbiology.

[18]  A. Maclean,et al.  Toxicity evaluation of replication-competent herpes simplex virus (ICP 34.5 null mutant 1716) in patients with recurrent malignant glioma , 2000, Gene Therapy.

[19]  R. Martuza,et al.  Conditionally replicating herpes simplex virus mutant, G207 for the treatment of malignant glioma: results of a phase I trial , 2000, Gene Therapy.

[20]  R. Martuza,et al.  Effect of prior exposure to herpes simplex virus 1 on viral vector-mediated tumor therapy in immunocompetent mice , 1999, Gene Therapy.

[21]  R. Martuza,et al.  Attenuated, Replication-Competent Herpes Simplex Virus Type 1 Mutant G207: Safety Evaluation of Intracerebral Injection in Nonhuman Primates , 1999, Journal of Virology.

[22]  B Roizman,et al.  Replication-competent, nonneuroinvasive genetically engineered herpes virus is highly effective in the treatment of therapy-resistant experimental human tumors. , 1999, Cancer research.

[23]  K. Tyler,et al.  A herpes simplex virus DNA polymerase mutation that specifically attenuates neurovirulence in mice. , 1998, Virology.

[24]  R. Ramakrishnan,et al.  Deletion of multiple immediate–early genes from herpes simplex virus reduces cytotoxicity and permits long-term gene expression in neurons , 1998, Gene Therapy.

[25]  R. Coffin,et al.  High efficiency gene transfer to the central nervous system of rodents and primates using herpes virus vectors lacking functional ICP27 and ICP34.5 , 1998, Gene Therapy.

[26]  A. Jacobs,et al.  Therapeutic efficiency and safety of a second-generation replication-conditional HSV1 vector for brain tumor gene therapy. , 1997, Human gene therapy.

[27]  J. Ilonen,et al.  Interferon‐γ Production in Antigen Specific T Cell Response: Quantitation of Specific mRNA and Secreted Protein , 1997, Scandinavian journal of immunology.

[28]  B. Roizman,et al.  The range and distribution of murine central nervous system cells infected with the gamma(1)34.5- mutant of herpes simplex virus 1 , 1997, Journal of virology.

[29]  B. Roizman,et al.  The γ134.5 protein of herpes simplex virus 1 complexes with protein phosphatase 1α to dephosphorylate the α subunit of the eukaryotic translation initiation factor 2 and preclude the shutoff of protein synthesis by double-stranded RNA-activated protein kinase , 1997 .

[30]  J. Trojanowski,et al.  Therapy of a Murine Model of Pediatric Brain Tumors using a Herpes Simplex Virus Type‐1 ICP34.5 Mutant and Demonstration of Viral Replication within the CNS , 1996, Journal of neuropathology and experimental neurology.

[31]  D S Latchman,et al.  Gene delivery to the central and peripheral nervous systems of mice using HSV1 ICP34.5 deletion mutant vectors. , 1996, Gene therapy.

[32]  B. Roizman,et al.  Phenotypic properties of herpes simplex virus 1 containing a derepressed open reading frame P gene , 1996, Journal of virology.

[33]  R. Martuza,et al.  Attenuated multi–mutated herpes simplex virus–1 for the treatment of malignant gliomas , 1995, Nature Medicine.

[34]  B. Roizman,et al.  The regulation of synthesis and properties of the protein product of open reading frame P of the herpes simplex virus 1 genome , 1995, Journal of virology.

[35]  D. Pfaff,et al.  Inflammatory effects of gene transfer into the CNS with defective HSV-1 vectors. , 1994, Gene therapy.

[36]  B. Roizman,et al.  Replication, establishment of latency, and induced reactivation of herpes simplex virus gamma 1 34.5 deletion mutants in rodent models. , 1993, The Journal of clinical investigation.

[37]  B. Roizman,et al.  Mapping of herpes simplex virus-1 neurovirulence to gamma 134.5, a gene nonessential for growth in culture. , 1990, Science.

[38]  P. Arstila,et al.  Herpes simplex virus detection by macroscopic reading after overnight incubation and immunoperoxidase staining , 1988, Journal of clinical microbiology.

[39]  J. Ilonen.  Lymphocyte blast transformation response of seropositive and seronegative subjects to herpes simplex, rubella, mumps and measles virus antigens. , 1979, Acta pathologica et microbiologica Scandinavica. Section C, Immunology.

[40]  J. Markert,et al.  Treatment of intracranial gliomas in immunocompetent mice using herpes simplex viruses that express murine interleukins , 1998, Gene Therapy.

[41]  J. Glorioso,et al.  Deletion of the S component inverted repeat sequence c' and the nonessential genes U(S)1 through U(S)5 from the herpes simplex virus type 1 genome substantially impairs productive viral infection in cell culture and pathogenesis in the rat central nervous system. , 1997, Journal of neurovirology.