Effect of immunosuppression on dengue virus infection in mice

Mean survival time following intracerebral inoculation of dengue virus was reduced and the titre of the virus in the brain of immunosuppressed mice was markedly increased. A single dose of cyclophosphamide given 24 h after dengue virus i.c. or i.p. substantially reduced the number of antibody forming cells in the spleen. Three doses of dengue virus, each followed by cyclophosphamide 24 h later, produced specific hyporesponsiveness to the dengue virus but not to a heterologous virus (Coxsackie B4), with a reduction in antibody forming cells in the spleen of such animals against dengue virus but not against Coxsackie B4 virus. Adoptive immunity by antiserum was abolished along with increased titres of the virus in the brain of immunosuppressed mice but the protection could be restored by a second dose of antiserum. Pre-treatment of mice with immune or normal spleen cells i.v. or reconstitution of immunosuppressed mice by such cells had no effect. Thus, humoral antibodies play a crucially important role in host defence mechanism in recovery of mice from primary dengue virus infection

Effect of dengue virus infection on Fc-receptor functions of mouse macrophages

Fc-receptor-mediated attachment and ingestion of opsonized sheep erythrocytes (EA) by the macrophages of spleen and peritoneal cavity were studied during dengue virus type 2 (DV) infection of Swiss albino mice. Following intracerebral inoculation, virus antigen could be demonstrated by immunofluorescence in the splenic macrophages from day 4 and in peritoneal macrophages from day 5 post-infection, with a higher number of positive cells discernible on the 7th and 8th days. The virus could be isolated from spleen tissue from day 5. The total number of cells was markedly reduced from day 4 onwards both in the spleen and peritoneal cavity. A loss in the capacity to attach and ingest EA was noticed, the lowest values of attachment index (AI) and phagocytic index (PI) being reached on day 4. At later periods the AI values increased markedly but continued to be significantly less than those in uninfected control mice. The PI values continued to be lower throughout. The dichotomy between the Fc-mediated attachment and ingestion may be a mechanism for prevention of virus infection of macrophages

Evidence for latency of Japanese encephalitis virus in T lymphocytes

Activation of latent Japanese encephalitis virus (JEV) in the spleen has been studied by co-cultivation with allogeneic or syngeneic cells. Activated virus was isolated by cocultivation from T lymphocytes of spleen, as shown by indirect immunofluorescence or by inoculation into mice. The B lymphocytes and macrophages of latently infected mice did not reactivate the virus. A higher proportion of Lyt 1 cells than Lyt 2 cells were harbouring JEV as shown by indirect immunofluorescence. The spleen cells from latently infected mice elicited the lymphoproliferative response but this was much lower than that observed in the controls. These findings suggest the establishment of latent JEV infection in T lymphocytes

Immune response to Japanese encephalitis virus in mother mice and their congenitally infected offspring

The immune response to Japanese encephalitis virus (JEV) was assessed in JEV-infected mice (mothers) and their offspring. The congenitally infected baby mice responded poorly in all assays for cell-mediated immunity. The total number of their splenic cells remained unaltered but the percentage of T cells was significantly reduced; a depressed delayed hypersensitivity response was seen against both homologous (JEV) and heterologous (sheep erythrocytes) antigens. In addition, significantly higher leukocyte migration inhibition (LMI) of spleen cells in the presence of specific antigen was observed. Adult mice infected during pregnancy demonstrated an impaired delayed hypersensitivity response to JEV antigen only. LMI was positive in mothers at 2 weeks post-partum, but not at later periods

Transplacental Japanese encephalitis virus (JEV) infection in mice during consecutive pregnancies

Transplacental transmission of Japanese encephalitis virus (JEV) has been demonstrated in consecutive pregnancies of mice. Pregnant mice inoculated intraperitoneally with JEV transmit the virus to the foetus. When such female mice were mated again after 6 months, the virus could be isolated from the foetuses of the ensuing pregnancy. The incidence of abortion was increased significantly though the neonatal deaths were considerably less than during the first pregnancy. Intra-uterine infection occurred in spite of the presence of HAI antibodies against JEV in the preconceptional sera of the mice. The findings of the present study indicate the value of such a system for further investigations of the pathogenesis of JEV infection during pregnancy in humans

Breakdown of the blood-brain barrier during dengue virus infection of mice

A breakdown of the blood-brain barrier occurred in mice inoculated intracerebrally (i.c.) or intraperitoneally (i.p.) with dengue virus type 2 (DEN2). This resulted in leakage of protein-bound Evans blue dye and 51Cr-labelled erythrocytes into the brain tissue. The leakage increased with time after infection and coincided with an increase of a DEN2-induced cytokine, the cytotoxic factor (CF), in the spleens of such mice. The titres of virus in the brain increased exponentially in i.c. inoculated mice but the virus was not detected in brains of mice given DEN2 by the i.p. route. Similar breakdown of the blood-brain barrier also occurred in mice inoculated intravenously with CF; the damage was dose-dependent and the vascular integrity was restored during the 3 h period after inoculation. Treatment of mice with antihistamine drugs, blocking H1 or H2 receptors, decreased the DEN2-induced protein leakage by up to 50% in i.c. inoculated mice and up to 92% in those inoculated i.p. Indomethacin, a prostaglandin synthetase inhibitor, had no effect. In i.c. inoculated mice protein leakage was inhibited by about 60% by treatment with CF-specific (CFA) or DEN2-specific antisera (DEN2A) whereas protection was complete with the combined treatment with both antisera. On the other hand, in i.p. inoculated mice the inhibition of protein leakage was 80 to 89% with CFA. These findings show a breakdown of the blood-brain barrier leading to cerebral oedema during DEN2 infection which is mediated via the release of histamine by a virus-induced cytokine

Host defence mechanisms against dengue virus infection of mice

Serum obtained from mice 3 to 5 weeks after the third i.p. dose of dengue type 2 virus (DV) protected recipient mice against intracerebral challenge with DV, whereas the serum obtained after 1 and 2 weeks provided minimum protection. Adoptive intravenous transfer of immune spleen cells obtained from mice 1 to 5 weeks after immunization did not protect recipient mice against even a small dose (10 LD50) of DV. Depletion of T-cells by treatment of mice with anti-thymocyte serum did not potentiate DV infection. Development of a cell-mediated immune response (CMI) against DV was noted only at two periods by the leucocyte migration inhibition test (LMI), with borderline values of 20 and 21%. Dengue virus did not cause illness or death in mice when given by i.p. or i.v. routes and this was not affected by pre-treatment of mice with silica to damage local macrophages. It is concluded that humoral antibody plays a critical role in recovery from primary dengue virus infection of mice whereas CMI and macrophages appear to have no protective role

Japanese encephalitis virus latency following congenital infection in mice

Latent Japanese encephalitis virus (JEV) infection was shown in inapparently congenitally infected Swiss albino mice after their mothers had been given JEV intraperitoneally during pregnancy. Only one of 37 (2.7%) of the baby mice showed persistence of infectious virus at 5 weeks of age. Reactivation of JEV in Swiss albino mice was demonstrated by stimulation with allogeneic spleen cells from Parks strain mice at 21 weeks of age; reactivation was demonstrated in 41% of the inapparently infected mice. The spleen cells of congenitally infected mice had depressed [3H] thymidine uptake following stimulation with concanavalin A, and depressed ability to induce a graft-versus-host response