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Guinea pigs infected by the peripheral route with the XJ pathogenic strain of Junin virus showed viscerotropism mainly in reticulo-phagocytic rich organs. By immunofluorescence, heavy infection of reticular-phagocytic cells was demonstrated, supporting the leading role of these cell types. Absence of neurotropism was demonstrated by the inability to recover infectious virus, as well as the absence of antigens, immunoglobulins, or 3rd component of complement deposits, in cells, vessels, or meninges. The correlation between infectivity and antigen expression observed in organs, and the absence of evidence of immunopathologic mechanisms, strongly suggest a direct viral effect in these experimental conditions. The results show that infection of guinea pigs by the peripheral route is an adequate model for human Argentine hemorrhagic fever with the exception of central nervous system involvement. Comparisons are made with infections produced in guinea pigs by attenuated strains, as well as with the disease in primates and humans.

Present knowledge points to horizontal transmission as the most significant mechanism for Junín virus maintenance in the main natural reservoirs, namely Calomys musculinus and Calomys laucha. The existence of naturally infected Akodon azarae, both within and outside the endemic area, as well as the finding that other species, ecologically and phylogenetically related to the main reservoirs, such as Akodon molinae and Calomys callidus, can experimentally develop persistent infections with virus shedding through fauces, suggest a potential role for these cricetids as alternative reservoirs. Furthermore, since those rodents inhabit the borders of the area in which Argentine Hemorrhagic fever is endemic, the risk of spread of this area is to be expected. Whether the establishment of Junín virus persistence in Calomys musculinus and other reservoirs depends on viral or host factors, such as a selective defect in L3T4+ lymphocytes as recently shown for mice, remains to be explored.

To assess the usefulness of the South American primate Cebus apella as a model for neurovirulence of Junín virus, eight monkeys were inoculated with 10(5) LD50 of the attenuated XJ-Clone 3 Junín virus strain by the intrathalamic route. After the second week, weight loss and polyadenopathies were observed in most animals, one-half of which had a transient leukothrombocytopenia. Moderate clinical central nervous system (CNS) involvement was present in four of eight monkeys, while the rest had only mild neurologic signs. All recovered except one, which developed a deep coma and was killed in a pre-mortem stage at 18 days post-infection (pi). Junín virus was isolated from the throat from five, from the blood from three, and from the brain from two monkeys. In the most severely ill animal, virus titers higher than viremia were detected in both inoculated and contralateral brain hemispheres, as well as in lung, lymph node, and small intestine. Junín antigens and "in vivo" bound immunoglobulins were detected by immunofluorescence (IF) in the brain of four animals at 18, 21, 40, and 155 days pi. Moderate lymphocytic parenchymal and meningeal infiltration were observed in the brain of four animals, and gliosis was also present in the most affected monkey. Although the clinical response to infection was not uniform, all infected monkeys developed high IF antibodies. Cebus apella cannot be used as a highly sensitive model for Argentine hemorrhagic fever (AHF). However, the results obtained show that the XJ-Clone 3 strain can replicate in the primate CNS and to induce lesions and immunoglobulin deposition. In addition, viral persistence is suggested by the late detection of viral antigens in brain at 40 and 155 days pi.

The present study investigated whether C. callidus, a species belonging to the Calomys genus, is capable of developing experimentally a persistent Junin virus (JV) infection. Newborn and adult cricetids were inoculated with the attenuated XJ-Clone 3 strain of JV by intracerebral or mucosal route. The present results indicate that the species is susceptible to JV infection, capable of shedding virus chronically through saliva and developing a persistent infection as shown by the detection of virus in brain tissue at 60 days post infection. These findings, and the fact that this cricetid shares its distribution areas with Calomys musculinus and Akodon azarae, support C. callidus as a potential JV reservoir.

The frequency of chromosomal aberrations in bone marrow cells of guinea pigs inoculated with the pathogenic XJ strain of Junin virus increased significantly at 6, 9, and 11 days postinoculation (p.i.). Animals inoculated with the attenuated XJ-clone 3 strain only showed significant increments of achromatic lesions (gaps) at 9 days p.i. Guinea pigs inoculated with the XJ-clone 3 strain and then treated with two doses of caffeine 24 and 12 h before killing at 9 days p.i. exhibited a significant increase of chromatid breaks and a parallel decrease of gaps. Because caffeine acts as an inhibitor of repair mechanisms of genetic damage, these results suggest a mutagenic effect of the attenuated strain.

Two Cebus sp surviving acute Junin virus infection, one after intramuscular inoculation with pathogenic XJ and the other after intracerebral inoculation with XJ Clone 3, failed to exhibit persistent infection. Although treatment with immunosuppressive drugs was carried out, no Junin virus was detected in blood or organs in spite of blind passages in mice for the former as well as cocultivation with permissive Vero cells for the latter. Viremia was also ruled out by immunofluorescence on BHK/21 cell culture. These findings correlate with the lack of long-lasting virus in blood observed in humans following the acute phase of Argentine Hemorrhagic Fever.

The usefulness of a method for detection of antibodies against Junin virus in whole blood was tested. N: NIH adult mice were inoculated with 10(3) PFU of attenuated XJ-Clon 3 Junin virus strain by intraperitoneal route and blood was obtained by retro-orbital puncture at 21 days post-infection. One blood aliquot (50 microliters) was collected in tubes containing a stabilizer solution for whole blood and another was processed for serum obtention. Immunofluorescent antibodies were tested on spot slides of a BHK/21 cell line persistently infected with Junin virus. High antibody titers (1/64 to 1/256) were detected in both whole blood and serum, with 66% coincidence between both procedures. These results show that the method of detection of antibodies in whole blood would be useful to test quickly for anti-Junin virus antibodies in seroepidemiologic studies, in endemic areas.

Experimental infection with the XJ-Clone 3 strain of Junín virus in laboratory bred Akodon molinae, a cricetid rodent inhabiting the borders of endemic Argentine hemorrhagic fever areas, was studied. Suckling animals inoculated intracerebrally proved sensitive and became chronically infected. Sixty percent of the rodents showed neurologic involvement, with mortality reaching 60%. Virus was recovered from the brain at 7, 15, 21, 37, and 57 days postinfection (pi). By immunofluorescence (IF), viral antigens were observed up to 182 days pi in cells of the central nervous system (CNS). Concurrently, immunoglobulin deposits were detected in infected CNS cells from 21 up to 182 days pi. These deposits increased with the progression of the immune response as measured by IF antibodies. The detection of immune complexes in brain cells of apparently healthy animals suggests that neither viral replication nor the development of a humoral immune response are necessary requisites for neurovirulence in this host. Infection of adult rodents by different routes failed to induce disease or mortality and virus could not be recovered from oral swabs, blood, or organs. Our data suggest that Akodon molinae could play a role in nature as an alternative reservoir of Junín virus in addition to the main reservoir, Calomys musculinus.