High resolution in situ hybridization to determine the cellular distribution of lymphocytic choriomeningitis virus RNA in the tissues of persistently infected mice: relevance to arenavirus disease and mechanisms of viral persistence
By the application of in situ hybridization to thin sections of paraffin-embedded tissues we have been able to determine with high resolution the cell types containing lymphocytic choriomeningitis virus nucleic acid in the tissues of persistently infected mice. We confirm and extend previous observations of virus persistence in the brain, lung, liver, kidney, pancreas, thyroid and reticuloendothelial system. In addition, we demonstrate for the first time persistence of viral nucleic acid in specific cell types in the thymus, lymph nodes, testes and bladder, and the adrenal, parathyroid and salivary glands; the cell types infected were observed in several animals. In lymphoid tissue, viral nucleic acid was predominantly located in the T cell-dependent areas of the spleen and lymph nodes; it was also present in cells of the thymic medulla. This has important implications for the deficiency in T cell function observed in persistently infected mice. In the testes, viral nucleic acid was detected in spermatogonia but not differentiating spermatocytes and therefore, in this tissue at least, persistence is related to the state of differentiation of the cell. Endocrine and exocrine dysfunctions have been described in persistently infected mice and we report that the highest levels of viral nucleic acid were found in the adrenal gland. The infection of endocrine and exocrine tissue was not pantropic, specific cell types expressed viral nucleic acid in each tissue. In the adrenal cortex, cells of the zona reticularis and zona fasciculata but not the zona glomerulosa were positive, whereas in the adrenal medulla viral nucleic acid was predominantly localized to adrenalin-secreting cells. Infection of the renal tubules, transitional epithelium of the bladder and the ducts of the salivary gland indicates the likely sites of virus production for the dissemination of arenavirus infections.
AhmedR.,
SalmiA.,
ButlerL. D.,
ChillerJ. M.,
OldstoneM. B. A.1984; Selection of genetic variants of lymphocytic choriomeningitis virus in spleens of persistently infected mice. Journal of Experimental Medicine 60:521–540
AhmedR.,
KingC.,
OldstoneM. B. A.1987; Virus-lymphocyte interaction: T cells of the helper subset are infected with lymphocytic choriomeningitis virus during persistent infection in vivo
. Journal of Virology 61:1571–1576
ArmstrongC.,
LillieR. D.1934; Experimental lymphocytic choriomeningitis of monkeys and mice produced by a virus encountered in studies of the 1933 St. Louis encephalitis epidemic. Public Health Report Washington 49:1019
BlountP.,
ElderJ.,
LipkinW. I.,
SouthernP. J.,
BuchmeierM. J.,
OldstoneM. B. A.1986; Dissecting the molecular anatomy of the nervous system: analysis of RNA and protein expression in whole body sections of laboratory animals. Brain Research 382:257–265
BuchmeierM. J.,
OldstoneM. B. A.1978; Virus-induced immune complex disease: identification of specific viral antigens and antibodies deposited in complexes during chronic lymphocytic choriomeningitis virus infection. Journal of Immunology 120:1297–1304
BuchmeierM. J.,
WelshR. M.,
DutkoF. J.,
OldstoneM. B. A.1978; The virology and immunobiology of lymphocytic choriomeningitis virus infection. Advances in Immunology 30:275–331
BuchmeierM. J.,
LewickiH. A.,
TomoriO.,
OldstoneM. B. A.1980; Monoclonal antibodies to lymphocytic choriomeningitis virus and Pichinde viruses: generation, characterization and crossreactivity with other arenaviruses. Virology 113:73–85
ByrneJ. A.,
OldstoneM. B. A.1984; Biology of cloned cytotoxic T lymphocytes specific for lymphocytic choriomengitis virus: clearance of virus in vivo
. Journal of Virology 51:682–686
De RoodD. G.1983; Proliferation and diffentiation of undifferentiated spermatogonia in the mammalian testis. In Stem Cells. Their Identification and Characterisation pp. 89–117 Edited by
PottenC. S.
Edinburgh: Churchill Livingstone;
DoyleM. V.,
OldstoneM. B. A.1978; Interactions between viruses and lymphocytes. I. In vivo replication of lymphocytic choriomeningitis virus during both chronic and acute viral infections. Journal of Immunology 121:1262–1269
DutkoF. J.,
OldstoneM. B. A.1983; Genomic and biological variation among commonly used lymphocytic choriomeningitis virus strains. Journal of General Virology 64:1689–1698
DymM.,
FawcettD. W.1971; Further observations on the numbers of spermatogonia, spermatocytes, and spermatids connected by intercellular bridges in the mammalian testis. Biology of Reproduction 4:195
GildenD. H.,
ColeG. A.,
MonjanA. A.,
NathansonN.1972; Immunopathogenesis of acute central nervous system disease produced by lymphocytic choriomeningitis virus. I. Cyclophosphamide-mediated induction of the virus carrier-state in adult mice. Journal of Experimental Medicine 135:860–873
HotchinJ.1962; The biology of lymphocytic choriomeningitis infection: virus-induced immune disease. Cold Spring Harbor Symposia on Quantitative Biology 27:479
HotchinJ. E.,
CinitsM.1958; Lymphocytic choriomeningitis infection of mice as a model for the study of latent virus infection. Canadian Journal of Microbiology 4:149–163
HuckinsC.1978; Spermatogonial intercellular bridges in whole-mounted seminiferous tubules from normal and irradiated rodent testes. American Journal of Anatomy 153:97
JamiesonB.,
AhmedR.1988; T cell tolerance: exposure to virus in utero does not cause a permanent deletion of specific T-cells. Proceedings of the National Academy of Sciences, U,. S,. A 85:2265–2268
KlavinskisL.,
OldstoneM. B. A.1987a; Lymphocytic choriomeningitis virus can persistently infect thyroid epithelial cells and perturb thyroid hormone production. Journal of General Virology 68:1867–1873
KlavinskisL.,
OldstoneM. B. A.1987b; Perturbation of host cell function during persistent virus infection. In Medical Virology vol VI pp. 85–98 Edited by
De La MazaM. L.,
PetersonE. M.
Amsterdam: Elsevier;
LipkinW. I.,
BattenbergE. L. F.,
BloomF. E.,
OldstoneM. B. A.1988; Viral infection of neurons can depress neurotransmitter mRNA levels without histologic injury. Brain Research 451:333–339
LipkinW. I.,
VillarrealL. P.,
OldstoneM. B. A.1989; Whole animal section in situ hybridisation and protein blotting: new tools in molecular analysis of animal models for human disease. Current Topics in Microbiology and Immunology 143:33–87
MimsC. A.1966; Immunofluorescence study of the carrier state and mechanism of vertical transmission in lymphocytic choriomeningitis virus infection in mice. Journal of Pathology and Bacteriology 91:395
MimsC. A.,
BlandenR. V.1972; Antiviral action of immune lymphocytes in mice infected with lymphocytic choriomeningitis virus. Infection and Immunity 6:695–698
MimsC. A.,
SubrahmanyanT. P.1966; Immunofluorescence study of the mechanism of resistance to superinfection in mice carrying the lymphocytic choriomeningitis virus. Journal of Pathology and Bacteriology 91:403
MoskophidisD.,
LohlerJ.,
Lehmann-GrubeF.1987; Antiviral antibody-producing cells in parenchymatous organs during persistent virus infection. Journal of Experimental Medicine 165:705–719
NathansonN.,
MonjanA. A.,
PanitchH. S.,
JohnsonE. D.,
PeturssonG.,
ColeG. A.1975; Virus-induced cell-mediated immunopathological disease. In Viral Immunology and Immunopathology pp. 357–391 Edited by
NotkinsA. L.
New York: Academic Press;
OldstoneM. B. A.,
RodriguezM.,
DaughadayW. H.,
LampertP. W.1984a; Viral perturbation of endocrine function: disordered cell function leads to disturbed homeostasis and disease. Nature, London 307:278–281
OldstoneM. B. A.,
SouthernP.,
RodriguezM.,
LampertP.1984b; Virus persists in B cells of islets of Langerhans and is associated with chemical manifestations of diabetes. Science 224:1440–1443
OldstoneM. B. A.,
AhmedR.,
ByrneJ.,
BuchmeierM. J.,
RiviereY.,
SouthernP.1985a; Virus and immune responses: lymphocytic choriomeningitis virus as a prototype model of viral pathogenesis. British Medical Bulletin 41:70–74
OldstoneM. B. A.,
AhmedR.,
BuchmeierM. J.,
BlountP.,
TishonA.1985b; Perturbation of differentiated functions during viral infection in vivo
. Virology 142:158–174
OldstoneM. B. A.,
BlountP.,
SouthernP. J.1986; Cytoimmunotherapy for persistent virus infection reveals a unique clearance pattern from the central nervous system. Nature, London 321:239–243
OldstoneM. B. A.,
SalvatoM.,
TishonA.,
LewickiH.1988; Biologic parameters of a virus variant that fails to generate CTL and establishes persistent infection in immunocompetent hosts. Virology 164:507–516
Reynolds-KohlerC.,
NelsonJ. A.1990; Techniques for doublelabelling virus infected cells. In Viral Pathogenesis: A Practical Approach Edited by
OldstoneM. B. A.
Oxford: IRL Press;
RodriguezM.,
BuchmeierM. J.,
OldstoneM. B. A.,
LampertP. W.1983; Ultrastructural localization of viral antigens in the CNS of mice persistently infected with lymphocytic choriomeningitis virus (LCMV). American Journal of Pathology 110:95–100
RodriguezM.,
GarrettR. S.,
RaittM.,
LampertP. W.,
OldstoneM. B. A.1985; Virus persists in beta cells of islets of Langerhans and infection is associated with chemical manifestations of diabetes. American Journal of Pathology 121:497–504
SalvatoM.,
ShimomayeE.,
SouthernP.,
OldstoneM. B. A.1988; Molecular characterization of LCMV Armstrong (CTL+) small genomic segment and that of its variant, clone 13 (CTL–). Virology 164:517–522
SouthernP. J.,
SinghM. K.,
RiviereY.,
JacobyD. R.,
BuchmeierM. J.,
OldstoneM. B. A.1987; Molecular characterization of the genomic S RNA segment from lymphocytic choriomeningitis virus. Virology 157:145–155
TaitS. A. S.,
OkamotoM.,
FloodC.,
TaitJ. F.1970; Production of steroids by in vitro superfusion of endocrine tissue. II. Steroid output from bisected whole, capsular and decapsulated adrenals of normal intact, hypophysectomized and hypophysectomized–nephrectomized rats as a function of time of superfusion. Endocrinology 86:360–381
TishonA.,
OldstoneM. B. A.1987; Persistent virus infection associated with chemical manifestations of diabetes. American Journal of Pathology 126:61–72
TraubE.,
KestingF.1964; Experiments on heterologous and homologous interference in LCM-infected congenitally with LCM virus. Zentralblatt für Veterinärmedizin 14:55
YatesR. D.,
WoodJ. G.,
DuncanD.1962; Phase and electron microscopic observations on two cell types in the adrenal medulla of the Syrian hamster. Texas Reports on Biology and Medicine 20:494–502
High resolution in situ hybridization to determine the cellular distribution of lymphocytic choriomeningitis virus RNA in the tissues of persistently infected mice: relevance to arenavirus disease and mechanisms of viral persistence