Physicochemical, biological and serological properties of a leukemogenic virus isolated from cultured RadLV-induced lymphomas of C57BL/Ka mice.

RadLV/VL₃, a virus spontaneously and continuously produced at high titer by BL/VL₃ cells, a line established in culture from a RadLV-induced C57BL/Ka mouse thymic lymphoma, has been extensively characterized. Evidence is presented indicating that it is a C-type RNA virus with highly thymotropic and leukemogenic biologic properties identical with those of its RadLV parent. It was not significantly neutralized by antibodies prepared against other ecotropic or xenotropic murine C-type viruses, used individually or in combination, whereas these antibodies effectively neutralized MCF-247 virus. However, antisera prepared against RadLV/VL₃ in rabbits and mice readily neutralized RadLV/VL₃. Curiously, the rabbit antiserum also had strong neutralizing activity against three other C-type endogenous viruses of the C57BL/Ka strain, BL/Ka(B), BL/Ka(N), and BL/Ka(X), all of which are fibrotropic (replicate readily in fibroblasts of appropriate tropism) and lack thymotropic and leukemogenic activity. In interference assays, these viruses and RadLV/VL₃ exhibited little or no capacity for mutual or self-interference, nor did RadLV/VL₃ interfere with the replication of MCF-247 virus in mink cells. However, the addition to cultures of increasing concentrations of the purified envelope glycoprotein (gp71) of RadLV/VL₃, though devoid of effect on BL/Ka(B) and BL/Ka(N), strongly inhibited the replication of RadLV and RadLV/VL₃ both in vitro and in vivo, and partially inhibited the xenotropic replication of BL/Ka(X). Competition radioimmunoassays with purified virion polypeptides demonstrated that the gp71 of RadLV/VL₃ is distinctively different from that of all other viruses tested; that the p12's of RadLV/VL₃ and BL/Ka(B) are similar or identical, whereas that of BL/Ka(N) competes in the AKR-MuLV p12 assay; and that the p30's of all of the viruses studied are indistinguishable by this method. Finally, RadLV/VL₃ failed to elicit cytopathic changes or transformed cell foci after serial passage in mink lung cells, even after co-infection with the xenotropic BL/Ka(X) virus, indicating that it is apparently devoid of MCF activity. Thus, MCF activity is apparently not a prerequisite for thymotropic and leukemogenic activity, at least in the C57BL/Ka strain.     

Nucleotide sequence of the envelope gene of radiation leukemia virus

The nucleotide sequence and the predicted amino acid sequence of the envelope gene (env) of RadLV/VL3 (T+L+) has been determined. RadLV/VL3 (T+L+) is a highly thymotropic (T+) and leukemogenic (L+) murine recombinant retrovirus continuously produced at high titer by BL/VL3 cells, a line established in culture from a radiation leukemia virus (RadLV)-induced C57BL/Ka mouse thymic lymphoma. The envelope gene is strikingly similar (92% homologous) to that encoded by the ecotropic Akv-MuLV. Among the differences, 134 scattered, although unevenly distributed point mutations (6.4%), two 9-bp deletions as well as a 6-bp insertion were observed (which result in 49 amino acid changes in the env gene of RadLV/VL3 (T+L+)).     

In vivo interaction between RNA viruses isolated from the C57BL/Ka strain of mice.

Virus replication can be initiated in the C57BL/Ka thymus by inoculation of three virus preparations isolated from mice of this strain, designated RadLV, RadLV-LTC, and BL/Ka(6), but not by three others, designated BL/Ka(B), BL/Ka(N), and BL/Ka(X). Coinfection by various mixtures of the three nonthymotropic viruses does not result in productive infection of the thymus. The addition of these nonthymotropic viruses to serial dilutions of RadLV, RadLV-LTC, or BL/Ka(6) does not significantly alter the replication of the wild-type RadLV preparations tested, whereas it significantly facilitates the replication of RadLV-LTC and BL/Ka(6). It is postulated that RadLV-LTC and BL/Ka(6) contain an attenuated or defective thymotropic and leukemogenic particle, the intrathymic replication of which is facilitated by coinfection with the nonthymotropic viruses.     

A new leukemogenic retrovirus isolated from tumor cells derived from a radio-induced lymphoma of C57BL/6 mice: analysis of the env and LTR sequences

We report the cloning and characterization of a new ecotropic provirus encountered in a radio-induced thymic lymphoma of the C57BL/6 mouse. The provirus with an abnormally long LTR was inserted in the chromosomal DNA within the Pvt-1/MLVi-1/Mis-1 region which is a common integration site for MCF virus in mice and for Mo-MuLV in rats. This new ecotropic provirus was molecularly cloned and found to be infectious and competent for replication after transfection of murine cells. The recovered virus termed T3651/B was B-ecotropic, T-lymphotropic (in vivo) and highly leukemogenic for newborn C57BL/6 mice and for adult mice provided they were submitted to a subleukemogenic dose of irradiation. As compared to the AKV prototype N-ecotropic endogenous retrovirus, the T3651/B env proteins are only affected by few scattered point mutations. In contrast, the LTR has five repeats of enhancer sequences containing consensus motifs specific of the nuclear factors NF1-like, LVa, LVb and SEF1. Since a virus with such properties was encountered only once in 31 radio-induced tumors and isolated at a fourth tumor passage, a direct role of T3651/B virus in tumor genesis after irradiation is uncertain. Nevertheless, it is clear that T3651/B virus is a new leukemogenic retrovirus with a particular LTR structure which fits well with the model proposed by Rassart et al. (J. Virol. 58, 96-106, 1986) for the emergence of a thymotropic highly leukemogenic RadLV.     

Nucleotide sequence of a radiation leukemia virus genome

The complete nucleotide sequence of an infectious molecular clone of a radiation murine leukemia proviral DNA RadLV/VL3(T+L+) has been determined. The sequence of the RNA genome is 8318 nucleotides long and contains three large open reading frames encoding the gag, pol, and env gene products. With the exception of a xenotropiclike R peptide and the LTR which bears structural similarities to a xenotropic LTR, displaying typical enhancerlike sequences, the remaining sequences are strikingly similar to the endogenous, ecotropic Akv murine leukemia virus. Therefore, it could be postulated that the leukemogenic properties of RadLV/VL3(T+L+) were generated by a recombination event between a xenotropic virus and an Akv-like ecotropic virus.     

Measurement of binding specificity between T cell lymphomas and murine leukemia viruses

We have previously reported the presence of receptors on radiation leukemia virus (RadLV)-induced thymomas and malignant thymocytes from AKR mice which specifically bind retrovirus produced by these T cell clones. These receptors have been shown to have specificity for virus reminiscent of an immune-specific receptor. Previous studies on T cell lymphoma binding to retroviruses have involved measurement of the interaction of labelled virus with cells using fluorescence-activated cell sorter (FACS) analysis (McGrath et al., J. Virol. (1978) 25, 923; McGrath and Weissman, Cell (1979) 17, 65; Weissman and McGrath, Curr. Top. Microbiol. Immunol. (1982) 98, 103). Here we report development of an assay for measuring lymphoma binding to virus, prepared as an immunoabsorbent adhered to a microtiter plate. Using this assay, we have shown that only T and not B cell lymphomas can bind to T cell-tropic viruses, and some cell lines have greatest specificity for homologous virus. The AKR-derived T cell lymphomas, SL3 and KKT-2, show greater specificity for leukemogenic AKR viruses, than for an AKR xenotropic virus or the recombinant AKR virus, MCF247. The RadLV-induced T cell lines, C6VL/1 and BL/VL3, have been found to bind cross-reactively to several different murine leukemia viruses (MuLVs). RadLV-induced T cell lymphomas do have greater specificity for their cognate retroviruses since free, homologous retrovirus can best block the interaction between cells and virus adhered to the wells of a microtiter plate. Cross-reactive interactions are more easily demonstrated by this assay, probably because low avidity interactions are stabilized as a result of the mode of virus presentation. Binding specificity for retroviral envelope determinants has been demonstrated using a rat anti-retroviral antiserum prepared as an F(ab)1 fragment. This antiserum can inhibit the interaction between the C6VL/1 thymoma and its RadLV virus. Specificity of this antibody for a gp70-like protein was confirmed by NaDodSO4-polyacrylamide gel electrophoresis (PAGE) and by loss of this activity after absorption of antibody on virus. Antibodies specific for RadLV/VL3 gp70 determinants can inhibit the interaction of C6VL/1 with RadLV/VL3 suggesting that cross-reactive binding to heterologous virus is also specific for a gp70 viral env determinant.     

Characterization of virus-specific cytotoxic T cell clones from allogeneic bone marrow chimeras

We established several H-2-restricted lymphocytic choriomeningitis virus (LCMV)-specific cytotoxic T cell clones from spleens of virus-primed C57BL/6 or C57BL/10 (H-2b) and B10.BR (H-2k) mice and from allogeneic C57BL/10----B10.BR and B10.BR----C57BL/10 bone marrow chimeras. Two T cell clones of H-2b origin and restricted to H-2b, 3 of H-2k origin and restricted to H-2k were compared with two clones each derived from the two types of chimeras. Their surface phenotype was found to be Lyt-2+, L3/T4- and KJ16-133+ (2 of 9). Clones from chimeras expressed bone marrow donor H-2 and are restricted to the recipient H-2. H-2k-restricted clones were all specific for Kk whereas all H-2b-restricted clones were specific for Db. These restriction specificities could be further defined by the blocking activity of various monoclonal anti-H-2 antibodies. Interestingly the anti-H-2Db antibodies blocked the restricted virus-specific killing activity of the clones derived B10.BR----C57BL/10 chimeras much more effectively than the activity of the clones derived from conventional H-2b mice. The various clones differed with respect to their fine specificity for LCMV strains. The 3 clones of conventional B10.BR origin only recognized LCMV-WE but not LCMV-Armstrong, Aggressive or Docile; H-2b-restricted conventional clones recognized target cells infected with all LCMV strains except LCMV-UBC-Docile; the T cell clones from the bone marrow chimeras recognized with one exception all LCMV strains tested.     

Role of T cells in resistance to Theiler's virus infection.

Intracerebral infection of C57BL/10SNJ mice with Theiler's virus results in acute encephalitis with subsequent virus clearance and absence of spinal cord demyelination. In contrast, infection of SJL/J mice results in acute encephalitis, virus persistence, and immune-mediated demyelination. These experiments examined the role of T-cell subsets in the in vivo immune response to Theiler's virus in resistant C57BL/10SNJ mice. Depletion of T-cell subsets with monoclonal antibodies (mAbs) directed at CD3 (pan-T-cell marker), CD4+ (class II-restricted) or CD8+ (class I-restricted) T cells resulted in increased frequency of paralysis and death as a result of acute encephalitis. Neuropathologic studies 10 days after infection demonstrated prominent necrosis, primarily in the pyramidal layer of hippocampus and in the thalamus of mice depleted of T-cell subsets. In immunosuppressed and infected C57BL/10SNJ mice, analysis of spinal cord sections 35 days after infection demonstrated small demyelinated lesions relatively devoid of inflammatory cells even though virus antigen could be detected by immunocytochemistry. Both CD4+ and CD8+ T cells are important in the resistance to infection with Theiler's virus in C57BL/10SNJ mice. However, subsequent spinal cord demyelination, to the extent observed in susceptible mice, depends on the presence of virus antigen persistence and a competent cellular immune response.     

Stimulation of Macrophages by Endotoxin Results in the Reactivation of a Persistent Herpes Simplex Virus Infection

Previous work has shown that splenic macrophages derived from herpes simplex virus (HSV)-resistant C57BL/6 mice undergo a persistent HSV infection which is characterized by the continuous release of infectious virus particles from a small subpopulation of infected cells. Treatment of persistently infected macrophages for 2 weeks with lipopolysaccharide (LPS) resulted in an increase of HSV yield and in virus-induced cytopathic effects. HSV was also reactivated by treatment of macrophage cultures with lipid A or tumour necrosis factor (TNF). Like macrophages of C57BL/6 origin, cells from LPS-hyporesponsive C3H/HeJ mice could be persistently infected with HSV. These cells were resistant to LPS-induced virus reactivation. The results show that macrophages derived from C57BL/6 mice are rendered susceptible to lytic HSV infection by treatment with LPS or TNF. Thus, these substances may interfere with persistent HSV infection which can be established due to genetically controlled properties of the host.     

A unique subset of normal murine CD4+ T cells lacking Thy-1 is expanded in a murine retrovirus-induced immunodeficiency syndrome, MAIDS

Some strains of mice inoculated with LP-BM5 murine leukemia virus (MuLV) develop a syndrome, termed mouse acquired immunodeficiency syndrome (MAIDS), characterized by progressive lymphoproliferation and profound immunodeficiency. LP-BM5 MuLV is a virus mixture that contains ecotropic (eco) and mink cell focus-induced MuLV and a defective genome that is the proximal cause of disease. Flow cytometry analyses of spleen and lymph nodes from susceptible C57BL/6 mice infected with this virus mixture revealed the presence in spleen and peripheral lymph nodes of a previously unrecognized subset of CD4+CD3+ T cells that are Thy-1-. The frequency of these cells increased with progression of disease, eventually comprising between 30% and 50% of all CD4+ cells. Infection of A/J mice, a strain which is genetically resistant to development of MAIDS, did not induce an increase of this T cell population, indicating that infection with the virus mixture was insufficient to induce its proliferation. A central role for the defective virus in this process was suggested by the finding that C57BL/6 mice infected with LP-BM5 eco alone did not have increased frequencies of Thy-1-CD4+ cells in spleen. Studies of spleen and peripheral lymph node cells from normal mice demonstrated the presence of Thy-1-CD4+ cells at frequencies of 1%-2%. Studies using two anti-T cell monoclonal antibodies, SM6C10 and SM3G11, that define four CD4+ subsets showed that Thy-1-CD4+ T cells from normal and infected mice were present only in the 6C10- subsets.     

Murine mammary tumor virus (MuMTV) infection of an epithelial cell line established from C57BL/6 mouse mammary glands.

An epithelioid cell line derived from the mammary glands of a C57BL/6 mouse and designated C57MG cell line was found to be susceptible to infection by routine mammary tumor virus (MuMTV). Although uninfected C57MG cells contain endogenous MuMTV-related DNA sequences, no RNA sequences homologous to MuMTV were detectable, even after treatment with the glucocorticoid, dexamethasone. However, after infection with MuMTV, these cells acquire additional MuMTV DNA, and viral RNA and proteins were readily detectable. Most, if not all, of the additional MuMTV DNA in infected C57MG cells appeared to be integrated. Synthesis of viral RNA and protein, and the release of virions into culture fluid by infected C57MG cells, was stimulated by incorporation of dexamethasone in the growth medium. The efficiency of infection by MuMTV in C57MG cells was similar to that in nonmurine cells. There was a direct relationship between multiplicity of infection (m.o.i.) and the average number of MuMTV RNA molecules detected per cell 5 weeks after infection. However, even at the highest m.o.i. used (4 × 10⁵ virions/cell), a plateau in the average number of viral RNA molecules per cell was not achieved. The origin of MuMTV synthesized by infected C57MG cells, whether it is the progeny of infecting MuMTV or of the endogenous C57BL/6 MuMTV or of both, remains undetermined. We have been unable to detect any morphological or growth pattern changes in C57MG cells following infection with MuMTV.     

Immunopathology of B-cell lymphomas induced in C57BL/6 mice by dualtropic murine leukemia virus (MuLV).

Combined clinicopathologic and immunomorphologic evidence is presented that would indicate that a murine leukemia virus (MuLV) with the dualtropic host range is capable of producing a clinically malignant lesion composed of immunoblasts and associated plasma cells in C57BL/6 mice. This process, morphologically diagnosed as an immunoblastic lymphoma of B cells using standard histopathologic criteria, was found to be distinctly polyclonal with regard to immunoglobulin (Ig) isotype when analyzed for both surface and cytoplasmic Ig. Further studies demonstrated that this clinicopathologically malignant, dualtropic MuLV-induced, polyclonal immunoblastic lymphoma of B cells in C57BL/6 mice was normal diploid and unable to be successfully transplanted to nonimmunosuppressed syngeneic recipients. Although all serum heavy and light chain components were found to be progressively elevated as the tumor load increased, the polyclonal increase in serum immunoglobulins was most pronounced for mu heavy and kappa light chains (ie, mu greater than gamma 2A greater than alpha greater than gamma 2B greater than gamma 1; kappa greater than lamba). The dissociation of clinicopathologic and biologic criteria for malignancy in the presently described dualtropic (RadLV) MuLV-induced B-cell lesion is sharply contrasted with the thymotropic (RadLV), MuLV-induced T-cell lymphoblastic lymphoma in C57BL/6 mice. This process is also a clinicopathologically malignant lesion but, when one uses biologic criteria, is found to be distinctly monoclonal, aneuploid, and easily transplanted to nonimmunosuppressed syngeneic recipients. The close clinicopathologic and biologic similarities of the dualtropic MuLV-induced animal model to corresponding human B-cell lymphoproliferative diseases are stressed.     

Either a CD4(+)or CD8(+)T cell function is sufficient for clearance of infectious virus from trigeminal ganglia and establishment of herpes simplex virus type 1 latency in mice

Following ocular infection of normal mice, herpes simplex virus type 1 (HSV-1) establishes a latent infection in the trigeminal ganglia (TG) with the complete absence of detectable infectious virus. In this study, the role of CD4(+)and CD8(+)T cell dependent immune responses is examined in relation to clearing infectious virus from the TG following HSV-1 ocular challenge. Nude mice, which lack T cells, and MHC(o/o)mice, which lack both MHC class I and MHC class II, were challenged ocularly with wild-type HSV-1. Over 70% of the TG from mice surviving the infection contained infectious virus, indicative of a chronic infection in these TG, rather than a latent infection. No infectious virus was detected in TGs from infected C57BL/6 parental mice. Ocular challenge of CD4(o/o)A(beta(o/o, CD8(o/o)or beta(2)m(o/o)mice resulted in latent rather than chronic infection. Similarly, when C57BL/6 mice were depleted for CD4(+)or CD8(+)T cells from 4 days before ocular challenge to 26 days after ocular challenge, no free virus was detected in TGs of challenged mice. In contrast, when mice were depleted of both their CD4(+)and CD8(+)T cells, over 90% of TGs were positive for free virus, suggesting that the lack of virus clearance was due to the combined lack of both CD4(+)T cells and CD8(+)T cells (i.e. in the presence of either CD4(+)T cells or CD8(+)T cells alone all of the infectious virus was cleared and latency was established).))Copyright 1999 Academic Press.     

IL-18, but not IL-12, is required for optimal cytokine production by influenza virus-specific CD8+ T cells

The potent innate cytokines IL-12 and IL-18 are considered to be important antigen-independent mediators of IFN-gamma production by NK cells and T lymphocytes. The present analysis addresses the physiological role of IL-12 and IL-18 in the generation of virus-specific CD8+ T cells. Both wt C57BL/6J (B6) mice and mice with disrupted IL-12p40 (IL-12p40(-/-)) or IL-18 (IL-18(-/-)) genes were infected with an influenza A virus and the characteristics of the resultant epitope-specific CD8+ T cell responses were compared. While IL-12 appeared to have no notable effect on either virus growth or on CD8+ T cell response profiles, the absence of IL-18 was associated with delayed virus clearance from the lung and, despite normal numbers, a significantly reduced production of IFN-gamma, TNF-alpha, and IL-2 by epitope-specific CD8+ T cells. While this cytokine phenotype was broadly maintained in IL-12p40/IL-18 double-knockout mice, no evidence was seen for any additive effect. Together, our results suggest that IL-18, but not IL-12, induces optimal, antigen-specific production of key cytokines by CD8+ T cells for the efficient clearance of influenza virus from the lungs of infected mice.     

Antibody response is required for protection from Theiler's virus-induced encephalitis in C57BL/6 mice in the absence of CD8+ T cells

Intracerebral infection of susceptible mice with Theiler's murine encephalomyelitis virus (TMEV) induces immune-mediated demyelinating disease and this system serves as a relevant infectious model for human multiple sclerosis. It was previously shown that beta2M-deficient C57BL/6 mice lacking functional CD8+ T cells display increased viral persistence and enhanced susceptibility to TMEV-induced demyelination, and yet the majority of mice are free of clinical signs. To understand the mechanisms involved in this general resistance of C57BL/6 mice in the absence of CTL responses, mice (muMT) deficient in the B-cell compartment lacking membrane IgM molecules were treated with anti-CD8 antibody and then infected with TMEV. Although little difference in the proliferative responses of peripheral T cells to UV-inactivated TMEV and the resistance to demyelinating disease was observed between virus-infected muMT and control B6 mice, the levels of CD4(+) T cells were higher in the CNS of muMT mice. However, after treatment with anti-CD8 antibody, 100% of the mice displayed clinical gray matter disease and prolonged viral persistence in muMT mice, while only 10% of B6 mice showed clinical symptoms and very low viral persistence. Transfusion of sera from TMEV-infected B6 mice into anti-CD8 antibody-treated muMT mice partially restored resistance to virus-induced encephalitis. These results indicate that the early anti-viral antibody response is also important in the protection from TMEV-induced encephalitis particularly in the absence of CD8+ T cells.     

Mousepox in inbred mice innately resistant or susceptible to lethal infection with ectromelia virus. IV. Studies with the Moscow strain

Summary The pathogenesis and transmission of infection with the Moscow strain of ectromelia virus were studied in inbred mice. BALB/cAnNcr had high morbidity and mortality and C57BL/6Ncr (B6) mice had high morbidity and low mortality. Virus was detected in B6 mice for 2 weeks after subcutaneous (s.c.) inoculation and infected mice developed lesions compatible with acute mousepox. B6 inoculated by footpad transmitted infection to cagemates for up to five weeks and soiled cages that had housed infected mice were infectious for three weeks. S.c.-inoculated B6 mice also transmitted by contact for 2 weeks. Transmission was attributed to oronasal excretion of virus. Airborne transmission of infection between adjacent cages occurred at a low rate. Ectromelia virus-free progeny were derived from previously infected dams. These studies indicate that the highly virulent and infectious Moscow strain of ectromelia virus caused self-limiting infection in inbred mice and that direct contact is the most efficient means of transmission. These findings support the concept that mousepox can be contained by husbandry practices that minimize or eliminate the spread of infection by direct contact or fomites.     

Cutaneous leishmaniasis: co-ordinate expression of granzyme A and lymphokines by CD4+ T cells from susceptible mice.

We have recently demonstrated that the frequency of T cells expressing granzyme A is significantly higher in skin lesions and spleens of susceptible BALB/c mice compared with resistant C57BL/6 mice infected with Leishmania major, a cause of human cutaneous leishmaniasis. In the present study, we have performed in vitro studies to characterize the subpopulation, the antigen responsiveness and the lymphokine production pattern of granzyme A-expressing T cells in L. major-infected mice. Using a limiting dilution system for functional analysis of selected T cells at the clonal level, we could show that granzyme A activity in infected BALB/c mice can be assigned to L. major-reactive CD4+ T cells secreting interleukin-2 (IL-2) and IL-4. Granzyme A production was most pronounced in the early phase of infection. On the other hand, granzyme A expression could not be detected in C57BL/6-derived T cells responding to L. major. The data support the suggestion that granzyme A is produced by L. major-responsive CD4+ T cells facilitating lesion formation and the dissemination of infection.     

Cytotoxic CD8+ T cell ablation enhances the capacity of regulatory T cells to delay viral elimination in Theiler's murine encephalomyelitis

Theiler's murine encephalomyelitis (TME) of susceptible mouse strains is a commonly used infectious animal model for multiple sclerosis. The study aim was to test the hypothesis whether cytotoxic T cell responses account for the limited impact of regulatory T cells on antiviral immunity in TME virus‐induced demyelinating disease (TMEV‐IDD) resistant C57BL/6 mice. TME virus‐infected C57BL/6 mice were treated with (i) interleukin‐2/‐anti‐interleukin‐2‐antibody‐complexes to expand regulatory T cells (“Treg‐expansion”), (ii) anti‐CD8‐antibodies to deplete cytotoxic T cells (“CD8‐depletion”) or (iii) with a combination of Treg‐expansion and CD8‐depletion (“combined treatment”) prior to infection. Results showed that “combined treatment”, but neither sole “Treg‐expansion” nor “CD8‐depletion,” leads to sustained hippocampal infection and virus spread to the spinal cord in C57BL/6 mice. Prolonged infection reduces myelin basic protein expression in the spinal cord together with increased accumulation of β‐amyloid precursor protein in axons, characteristic of myelin loss and axonal damage, respectively. Chronic spinal cord infection upon “combined treatment” was also associated with increased T and B cell recruitment, accumulation of CD107b⁺ microglia/macrophages and enhanced mRNA expression of interleukin (IL)‐1α, IL‐10 and tumor necrosis factor α. In conclusion, data revealed that the suppressive capacity of Treg on viral elimination is efficiently boosted by CD8‐depletion, which renders C57BL/6 mice susceptible to develop chronic neuroinfection and TMEV‐IDD.     

Role of interferon in persistent infection of macrophages with herpes simplex virus

Splenic macrophage cultures from C57BL/6 mice resistant to infection with herpes simplex virus (HSV) in vivo survived HSV infection in vitro. In contrast, macrophages from HSV-susceptible DBA/2 mice were completely lysed by the virus. During prolonged culturing, macrophages from C57BL/6 mice continued to produce infectious virus, indicating establishment of a persistent infection. At this time, interferon (IFN) was undetectable. However, as shown directly by the addition of an anti-IFN serum and indirectly by an increased activity of (2'-5')oligoadenylate synthetase, IFN was involved in the maintenance of the persistent infection. During the acute phase of virus infection, viral DNA replication was identical in macrophages from resistant or susceptible mice. Later, viral DNA content and the number of cells expressing HSV antigens decreased in macrophages from C57BL/6 mice. However, single cells remained to express viral proteins and to produce infectious particles. The results show that macrophages can be persistently infected with HSV due to their genetically controlled properties.