Eosinophilic Intracytoplasmic Inclusion Research Articles

Poxvirus in farmed Nile crocodiles.

In an outbreak of wart-like skin lesions in nine-month-old farmed Nile crocodiles (Crocodylus niloticus) the lesions were especially common on the head and neck but occurred on all parts of the body except the tail. Eosinophilic intracytoplasmic inclusion bodies were present in epithelial cells and further examination of the lesions by transmission electron microscopy revealed the presence of a poxvirus. An autogenous vaccine was produced from material from the lesions and in six vaccinated crocodiles the lesions healed more quickly than in unvaccinated controls.

Neurotoxicity of vincristine after the osmotic opening of the blood-brain barrier.

The direct effect of intravascularly injected vincristine on the rat brain was investigated after the osmotic opening of the blood-brain barrier. This was opened unilaterally by injecting 2.5 ml of 1.4 molal mannitol solution into the right carotid artery through a cannula inserted in the external carotid artery in retrograde fashion. Vincristine (0.75 mg/kg) was then injected into either the carotid artery or into a vein. Animals, thus treated, developed left hemiparesis and choreo-athetoid head motion within several days. Microscopical analysis revealed 1 intracytoplasmic eosinophilic inclusions at an early period, 2 neuronal argyrophilic change and axonal thickening with spheroid formation at later times, and 3 glial mitotic arrest, a finding not reported in the previous studies with intrathecal injection of vincristine. Neuronal changes were usually confined to the selected area in the midbrain, whereas glial mitotic figures were seen in the hippocampus, midbrain and cerebellum. These changes are thought to be direct effects of the vincristine binding with cellular microtubules, including mitotic spindle tubules and neurotubules. The transient opening of the blood-brain barrier seems to be a useful technique for studying the experimental neurotoxicology of drugs to which the blood-brain barrier is impermeable.

Ultrastructure of intracytoplasmic inclusion bodies in hepatocytes of griseofulvin-treated mice. Fibrils with myelin figure and rod-like fibrils.

Intracytoplasmic eosinophilic inclusion bodies light-microscopically identical to human alcoholic hyalin were produced in hepatocytes of griseofulvin-treated mice. Electron-microscopic investigation on these inclusion bodies revealed two morphologically different fibrils. The most frequently observed fibrils in liver tissues appeared straight or slightly curved in longitudinal sections and granular in cross sections. High-power electron micrographs on these fibrils revealed that each fibril consisted of parallel-running fine filaments of about 1.25 nm thick which were separated by spaces of about 1.25 nm. The ultrastructural appearance of these fibrils resembled that of myelin sheath in longitudinal sections. They usually measured from 13 to 32 nm wide and up to 0.9 micron long. Fibrils with larger diameter of up to 62 nm were infrequently encountered. Rod-like fibrils ultrastructurally similar to those seen in human alcoholic hyalin were also found in isolated materials. All these fibrils were isolated in the same fraction by a slight modification of a discontinuous sucrose gradient ultracentrifugation and a two phase polymer centrifugation. The present observation suggests that intracytoplasmic inclusion bodies produced in this animal model are composed of morphologically different fibrils. These different fibrils could share the same origin.

Reduced pathogenicity associated with a small plaque variant of the Egyptian strain of Rift Valley fever virus (ZH501)

Variants of Rift Valley fever virus producing plaques in CER cells of four different sizes are described. A plaqueforming unit (PFU) variant forming minute plaques was isolated and purified. Virus derived from this variant was not pathogenic to adult Swiss albino mice by the intraperitoneal (i.p.) route and was less pathogenic than the parent strain (ZH501) to adult Sprague Dawley rats by i.p. route, but produced typical severe liver necrosis in adult Syrian hamsters with intranuclear and intracytoplasmic eosinophilic inclusions. Antigen and antiserum to the minute variant prepared in mice reciprocally cross-reacted with antisera and antigens of the original strain (ZH501) in the complement fixation test. Plaque size of the minute variant remained constant after serial passages in cell culture and in suckling mouse brain. When the minute plaque variant was passaged i.p. in hamsters, virus which formed large plaques in CER cells was recovered from the hamster sera.

Granulofilamentous inclusions in a meningioma.

We have studied the histology, immunocytochemistry, and ultrastructure of a syncytial meningioma which contained the unusual feature of large eosinophilic intracytoplasmic inclusions. Electron microscopic examination revealed that these bodies were granular, osmiophilic masses, closely associated with cytoplasmic filaments of intermediate size. Desmosomal junctions with a somewhat abnormal morphology were also observed, and the possible relationship between the inclusions and the dense components of desmosomes is discussed. This tumor is also compared to other tumors of the central nervous system with hyaline and granulofilamentous inclusions.

Intracytoplasmic neuronal inclusions in the human thalamus. Light-microscopic, histochemical, and ultrastructural observations.

Intracytoplasmic eosinophilic inclusions were found by light microscopy in the thalamic neurons of 35 consecutive normal adult brains and in a case of myotonic dystrophy, but not in six newborn children, including one with myotonic dystrophy. Histochemical tests suggested a protein composition. Ultrastructurally, the inclusions were composed of stacks of parallel alternating dark and light rectilinear profiles not surrounded by a limiting membrane. Such inclusions are a virtually constant finding in the adult human thalamus and probably represent sites of neuronal protein storage.

Alpha-1-antitrypsin in intracellular inclusions of diethylnitrosamine induced hepatomas of C57BLxC3H F1 mice.

Six large hepatic nodules obtained 46 to 74 weeks after a single dose of diethylnitrosamine (0.625 or 1.25 micrograms/gm) in infant C57BLxC3H F1 male mice were studied to elucidate the nature of intracytoplasmic inclusions in the hepatic cells of these nodules. By light microscopy, the nodules constituted a spectrum of lesions ranging from some which were well differentiated adenomas to others which were frank carcinomas. Round or oval eosinophilic intracytoplasmic inclusions were consistently seen in the well differentiated adenomatous lesions. Some of these inclusions were PAS positive and resisted diastase digestion while others were either negative or only weakly PAS positive. By electron microscopy, they were most commonly crystalline, contained a reticulated electron dense material and were located in the distended rough endoplasmic reticulum. On immunofluorescence the inclusions showed marked specific reactivity with antisera against human alpha-1-antitrypsin. The presence of alpha-1-antitrypsin in these inclusions was further supported by the resistance of these inclusions to digestion by trypsin or papain but not by the pepsin or pronase.

Canine distemper virus-associated encephalitis: modification by passive antibody administration.

Gnotobiotic puppies were intraperitoneally infected with R252-canine distemper virus (CDV), and treated with normal serum (NS) or CDV-specific antibody. Resultant central nervous system (CNS) lesions were evaluated by light and immunofluorescence microscopy. Lesions in NS-treated dogs, if present, consisted of small foci of neuronal degeneration and accompanying microgliosis, restricted to grey matter of the frontal lobe. Dogs receiving anti-CDV antibody, in contrast, had extensive lesions in the brain. Laminar cortical necrosis, gliosis and astrocytic activation of the grey matter of the frontal lobe was the most consistent lesion observed. In addition, foci of cerebellar granule cell layer degeneration and necrosis was seen along with segmental poliomalacia of the spinal cord. In several dogs, eosinophilic intracytoplasmic and intranuclear viral inclusion bodies were observe in neurons. The lesions observed in these dogs are compared to previously reported lesions of CDV infection in dogs.

Replication of measles virus in a cell culture from a glioblastoma of human origin.

A cell culture from a glioblastoma of human origin infected with the Edmonston strain of measles virus produced and released infectious measles virus. All cellular types seemed to be involved in the process of virus replication. Staining with hematin-eosin revealed the presence of eosinophilic intracytoplasmic and intranuclear inclusions. Examination with the electron microscope revealed viral nucleocapsids in the cytoplasm and, rarely, in the nuclei of infected cells.

The recognition of viral and chlamydial infection in cervical smears

In the 12 mth ended 30 June 1976, 14,868 cervical smears (from 13,212 patients) were examined in the Cytology Department, King George V Hospital, Sydney. Forty-two smears (1 in 354), from 37 patients, showed features of viral or chlamydial infection. Appearances were considered diagnostic in 21 smears and suggestive of infection in a further 21 smears. Changes were seen which were attributable to herpes virus, the virus of condyloma acuminatum, cytomegalovirus, adenovirus and TRIC agents. Six smears showed changes consistent with infection by virus other than those mentioned. The most characteristic feature of viral and chlamydial infection is the presence of inclusion bodies in endocervical and metaplastic squamous cells. Intranuclear inclusions are seen in herpes (where the cells are often multinucleate), cytomegalovirus and adenovirus infection. These are eosinophilic in herpes and adenovirus and basophilic in cytomegalovirus infection. Eosinophilic intracytoplasmic inclusions are seen in TRIC agent infection and sometimes in cytomegalovirus infection. Recognition is important for several reasons. Cytologically, changes due to viral and chalmydial infection may be present in association with or may also to some extent simulate precancerous or cancerous changes. Epidemiologists have considered the possibility that a sexually transmitted viral carcinogen is important in the aetiology of cervical cancer. Clinically infection may be asymptomatic or be responsible for severe, even necrotizing, cervicitis. The presence of herpes, cytomegalovirus and TRIC agents in the genital tract in early pregnancy is associated with abortion, prematurity and dysmaturity. Some obstetricians consider the presence of herpes or cytomegalovirus in late pregnancy an indication for caesarean section before membrane rupture in order to prevent serious, and sometimes fatal, infection of the neonate occurring during delivery through an infected birth canal.

Isolation and Characterization of a Lymphocyte Associated Foamy Virus from a Red Uakari Monkey

A syncytium-forming virus was isolated from the lymphocytes of a uakari monkey 70 days after establishing a lymphocyte/owl monkey kidney coculture in the wild. The morphology and morphogenesis of the virus, plus its physicochemical characteristics indicate that the agent is a foamy virus. An unusal cell alteration found in cultures infected with this foamy was the formation of eosinophilic intracytoplasmic inclusion in association with the syncytia. The agent was found to be antigenically distinct to other known simian foamy viruses.

Isolation of respiratory syncytial virus from calves with acute respiratory disease.

Viruses were recovered from nasal exudates of two of nine calves with acute respiratory disease. The strains, designated VC484 and VC488, were neutralized markedly by guinea pig antiserum against the Long strain of respiratory syncytial (RS) virus. Physical and chemical properties of the isolates were characteristic of RS viruses. Cultures of infected primary bovine embryonic kidney cells developed numerous syncytial cells. Intracytoplasmic eosinophilic inclusions were present in syncytial and single cells. Infected cell cultures did not adsorb guinea pig erythrocytes, nor did infective culture fluids agglutinate guinea pig, rat, mouse, rabbit, calf, or horse erythrocytes. One of the two calves developed a significant increase in titer of antibody; the other calf had preexisting neutralizing antibody against both isolates. No other viruses were recovered. It is concluded that these calves were infected with bovine strains of RS virus, reported now in the United States.

Isolation and characterization of a cell line from the cocultivation of Lucké renal tumor cells and nontransformed feeder cells.

The in vitro cocultivation of renal adenocarcinoma (Lucké tumor) cells with a feeder layer of frog tongue cells at 28° C successfully yielded 2 newly established cell lines. Data on the morphology,growth characteristics, and karyology of these cells strongly suggested that the established cell lines (LT-1 and LT-2) originated from the Lucké tumor. LT-1 cells transplanted into the anterior eye chamber of Rana pipiens proliferated like transformed or neoplastic cells. Eosinophilic and Feulgenpositive intracytoplasmic inclusion bodies were detected in LT-1 cells. Electron microscopic studies showed that inclusion bodies contained cell components and irregular structures of varying sizes and shapes but no intact Lucké herpesvirus (LHV) particles. Treatment of LT-1 cells with 5-bromodeoxyuridine and 5-iododeoxyuridine did not induce LHV multiplication.

A post-vaccinal inclusion body encephalitis in dogs.

Two separate episodes of an encephalitis occurred in dogs after the administration of a few batches of a combined, live attenuated distemper/hepatitis virus vaccine. The lesions consisted of eosinophilic intranuclear and intracytoplasmic inclusions in large neurones, together with a nonsuppurative encephalitis and malacia of the ventral pontine gray matter. The presence of nucleocapsid, indistinguishable from that seen in distemper infection, indicates that a distemper or distemper-like virus was directly involved.

Sealpox field survey.

The light microscopic demonstration of eosinophilic intracytoplasmic inclusion bodies in epithelial cells and the electron microscopic demonstration of poxvirus in these inclusions has shown that sealpox occurs in free-living California sea lions, Zalophus calijornianus. This is the first evidence of a poxvirus skin infection in free-living California sea lions.

Production of kidney tissue cultures from African green monkeys, experimentally infected with the causative agent of Frankfurt-Marburg-syndrome.

Green monkeys (Cercopithecus aethiops), experimentally infected with the causative agent of the Frankfurt-Marburg-Syndrome (FMS-virus), were killed after the appearance of disease symptoms, and. nephrectomized. Tissue cultures were set up from the trypsinized kidney tissue. Examination of live tissue culture under low magnification revealed no difference in rate of growth or appearance from the cultures of kidney cells from non-infected monkeys. Intracytoplasmic eosino-philic inclusion bodies could be found invariably in stained preparations of the cells of kidney tissue cultures deriving from infected monkeys, virus particles could be seen by electron microscopy, and viral antigen could be demonstrated in the cells with immunofluorescent antibody technique. That the tissue culture fluids contained infectious virus could be demonstrated by inoculating monkey kidney cell tissue cultures in stationary growth phase and observing the cyto-pathic effect.

パラインフルエンザウイルス (HVJ) のPS細胞における増殖と持続感染に関する研究

Myxovirus parainfluenzae 1 (HVJ) multiplied readily and could be passaged serially in the cells of an established strain, PS cell. The viral growth in PS cells was recognized easily by marked cytopathic changes, production of hemagglutinin, and positive cyto-hemadsorption. The infectivity titers of HVJ in PS cell were almost the same as EID50 titers, even when the egg-adapted virus was tested. It was concluded that PS cell was one of the most suitable host systems for HVJ.The intracellular development of viral antigen in HVJ-infected PS cells was studied by the fluorescent antibody technique. Specific viral antigen was first found as fine granules in the perinuclear area. The antigen steadily increased in quantity and was observed as numerous granules distributed throughout the cytoplasm. Later, the fluorescent material began to form larger aggregates and, in some cases, such large masses occupied most of the cytoplasm. No fluorescent material was found in the nucleus at any stage of the infection. Eosinophilic intracytoplasmic inclusion bodies were found in the cells infected with HVJ. The nature of this inclusion body is now under investigation.Viral carrier culture was obtained by prolonged cultivation of the cells which had survived HVJ-infection. The culture continued to produce virus over a period of many months. Immunofluorescent staining revealed that about 30% of cells in the culture appeared to contain viral antigen. Varying amount of viral antigen was contained in each of these cells. Thus, the cells which possessed very small amount of antigen as fine granules and the cells which contained large masses of fluorescent material were found simultaneously in the same culture. The nature of this viral carrier state was discussed in the light of the results obtained.

Studies on the causative agent of an epizootic amongst Manx shearwaters (Puffinus P. puffinus).

Evidence for the isolation of a virus from this epizootic disease of shearwaters is based upon the reproduction of a similar condition in ducklings and pigeons and upon the development of characteristic histological changes with cytoplasmic eosinophilic inclusions in inoculated chorio-allantoic membranes of chick embryos.It was not possible to reproduce the original disease exactly by inoculation of shearwaters with egg-passage virus from the previous season because the birds did not develop blisters at the site of inoculation of the web. Nevertheless, three out of four birds died and the agent was re-isolated from the spleen of one of them.The ability of the virus to cause blisters on duckling webs was neutralized by shearwater serum taken at the convalescent but not at the acute stage of the disease, and complement fixation was obtained with convalescent shearwater serum.Previous studies on natural virus infections of birds have been mainly limited to psittacosis and ornithosis, the pox viruses, Newcastle disease virus and some of the neurotropic viruses. However, the failure to infect mice by any route, and the ability of the shearwater virus to pass through filters with an A.P.D. of 48 mμ, seemed to rule out the psittacosis group as a cause of the infection. The ability of the virus to cause vesicles and the intracytoplasmic eosinophilic inclusion bodies suggested a member of the pox group of viruses, but the small size of the agent renders a relation to either the pox group or Newcastle disease virus unlikely. Several of the neurotropic viruses which infect birds, for example, Japanese B virus, are about the size of the shearwater virus, but they do not cause the formation of inclusion bodies and they are all pathogenic for mice. Foot and mouth disease virus is also of small size and produces vesicular lesions, but it is unlikely that the shearwater virus is the same since it was unable to infect a cow or the pad of a guinea-pig, and it grew in eggs more readily than foot and mouth disease virus.There has obviously been insufficient work on the agent to postulate a new virus, but the properties described do not justify its classification in any of the known virus groups. The name puffinosis which was originally suggested still seems suitable for this disease of Puffinus puffinus puffinus.