Detection of JC virus DNA sequences and expression of the viral regulatory protein T-antigen in tumors of the central nervous system.

JC virus (JCV) is a neurotropic polyomavirus infecting greater than 70% of the human population worldwide during early childhood. Replication of JCV in brains of individuals with impaired immune systems results in the fatal demyelinating disease, progressive multifocal leukoencephalopathy (PML). Furthermore, JCV possesses an oncogenic potential and induces development of various neuroectodermal origin tumors including medulloblastomas and glioblastomas in experimental animals. The oncogenecity of JCV is attributed to the viral early gene product, T-antigen, which has the ability to associate with and functionally inactivate well-studied tumor suppressor proteins including p53 and pRB: The observations from laboratory animal experiments have provided a rationale for examining the presence of the JCV DNA sequence and expression of the viral oncogenic protein in human brain tumors. We have examined 85 clinical specimens from the United Kingdom, Greece, and the United States, representing various human brain tumors including oligodendroglioma, astrocytoma, pilocytic astrocytoma, oligoastrocytoma, anaplastic astrocytoma, anaplastic oligodendroglioma, glioblastoma multiforme, gliomatosis cerebri, gliosarcoma, ependymoma, and subependymoma, for their possible association with JCV. We performed gene amplification techniques using a pair of primers that recognize the JCV DNA sequence, and we demonstrated the presence of the viral early sequence in 49 (69%) of 71 samples. More importantly, our results from immunohistochemistry analysis revealed expression of JCV T-antigen in the nuclei of tumor cells in 28 (32.9%) of 85 tested samples. These observations, along with earlier in vitro and in vivo data on the transforming ability of this human neurotropic virus invite additional studies to re-evaluate the role of JCV in the pathogenesis of human brain tumors.

Aberrant localization of the neuronal class III beta-tubulin in astrocytomas.

BACKGROUND: The class III beta-tubulin isotype (betaIII) is widely regarded as a neuronal marker in development and neoplasia. In previous work, we have shown that the expression of betaIII in neuronal/neuroblastic tumors is differentiation dependent. In contrast, the aberrant localization of this isotype in certain nonneuronal neoplasms, such as epithelial neuroendocrine lung tumors, is associated with anaplastic potential. OBJECTIVE: To test the generality of this observation, we investigated the immunoreactivity profile of betaIII in astrocytomas. DESIGN: Sixty archival, surgically excised astrocytomas (8 pilocytic astrocytomas, WHO grade 1; 18 diffuse fibrillary astrocytomas, WHO grade 2; 4 anaplastic astrocytomas, WHO grade 3; and 30 glioblastomas, WHO grade 4), were studied by immunohistochemistry using anti-betaIII monoclonal (TuJ1) and polyclonal antibodies. A monoclonal antibody to Ki-67 nuclear antigen (NC-MM1) was used as a marker for cell proliferation. Antibodies to glial fibrillary acidic protein (GFAP) and BM89 synaptic vesicle antigen/synaptophysin were used as glial and neuronal markers, respectively. RESULTS: The betaIII immunoreactivity was significantly greater in high-grade astrocytomas (anaplastic astrocytomas and glioblastomas; median labeling index [MLI], 35%; interquartile range [IQR], 20%-47%) as compared with diffuse fibrillary astrocytomas (MLI, 4%; IQR, 0.2%-21%) (P <.0001) and was rarely detectable in pilocytic astrocytomas (MLI, 0%; IQR, 0%-0.5%) (P <.0001 vs high-grade astrocytomas; P <.01 vs diffuse fibrillary astrocytomas). A highly significant, grade-dependent relationship was observed between betaIII and Ki-67 labeling and malignancy, but this association was stronger for Ki-67 than for betaIII (betaIII, P <.006; Ki-67, P <.0001). There was co-localization of betaIII and GFAP in neoplastic astrocytes, but no BM89 synaptic vesicle antigen/synaptophysin staining was detected. CONCLUSIONS: In the context of astrocytic gliomas, betaIII immunoreactivity is associated with an ascending gradient of malignancy and thus may be a useful ancillary diagnostic marker. However, the significance of betaIII-positive phenotypes in diffuse fibrillary astrocytomas with respect to prognostic and predictive value requires further evaluation. Under certain neoplastic conditions, betaIII expression is not neuron specific, calling for a cautious interpretation of betaIII-positive phenotypes in brain tumors.

Reduced variance in the latency and amplitude of the fifth wave of auditory brain stem response after normalization for head size.

The value of the Auditory Brain Stem Response (ABR) in the assessment and detection of neurological disorders could be considerably enhanced if the normative standards of ABR characteristic parameters take into account all other systematic sources of variance. The present study attempts to take into account the influence of head size on latency and amplitude of the ABR components. We examined amplitude and latency as a function of head size in 40 normal male subjects (age 20-40 years). Significant negative correlation was found between amplitude and head radius. The experimental data were fitted using a theoretical curve of the potential on the surface of a three-concentric sphere model representing the human head. The fitted curve of amplitude versus radius can be applied to normative data in order to substantially reduce dispersion and consequently increase the diagnostic value of this parameter. Moreover, a substantial effect of radius on wave V latency was detected. Normalization of the latencies with reference to the head radius, assuming that the latter is proportional to the length of brain stem, resulted in a significant reduction in the standard deviation of these data as compared to the original.

Medial medullary infarction from fibrocartilaginous embolism to the anterior spinal artery.

A previously healthy young woman presented with sudden onset of quadriplegia, anesthesia below the C3 dermatome, respiratory paralysis, vertical nystagmus, ocular bobbing and cortical blindness. After partial resolution of the latter deficits, she remained quadriplegic, with a C3 level of anesthesia, and in respiratory paralysis until death from complications of a fulminant pulmonary infection. Autopsy disclosed bilateral infarctions of the medial aspect of the medulla and the upper cervical cord, in the distribution of the anterior spinal artery. Multiple sites of arterial occlusion by fibrocartilaginous material were found in branches of the anterior spinal artery, in correspondence with the sites of infarction. A review of the literature disclosed only 2 examples of medial medullary syndrome from embolism to the anterior spinal artery.

Pathology of an experimental extradural spinal T cell tumor.

Syngeneic mice injected intravenously with a T cell tumor line (line 13) induced by Gross' murine leukemia virus developed paraparesis and sensory loss below the midthoracic level 2 to 3 weeks after inoculation. Although signs of systemic disease coexisted, the animals survived through the development of the neurologic symptoms, and treatment with cytotoxic agents was not required. Pathologic study of the spinal cord and brain revealed tumoral infiltration of the meninges, confined to the extradural spaces, more markedly at spinal than cerebral levels. Equally severe infiltrates occurred in the paravertebral musculature. No leptomeningeal or parenchymal involvement was present, irrespective of the severity of the extradural infiltration. Marked bone marrow and visceral infiltration coexisted with central nervous system involvement. The topography of the extradural and muscular tumor cells collections related to the proximity of the involved bone marrow and areas of direct communication between these spaces were repeatedly identified. On the other hand, line 13 cells injected directly into the brain substance produced diffuse leptomeningeal tumoral infiltration without extradural involvement. These findings suggest that the pathogenesis of this model of spinal T cell tumor proliferation involves a first stage of bone marrow infiltration, followed by extradural involvement. This occurs by direct migration of bone marrow tumor cells through gaps in the vertebral bone. This model offers the opportunity for the study of malignancies that produce bone destruction as a mechanism for tumoral spread.

New experimental model of meningeal leukemia.

Meningeal leukemia is a common, often fatal complication of leukemia. Efforts to devise better prophylactic or therapeutic regimens have been limited by large gaps in knowledge regarding the pathogenesis of the process. This report describes a new experimental model of meningeal leukemia that should facilitate its study and fill some of the gaps. T-cell lymphomas, originally induced in C3H mice by Gross murine leukemia virus, were established as transplantable tumor lines in syngeneic mice. Three of ten tumor lines produced meningeal leukemia when injected i.v. into normal recipient animals. The others produced predominantly visceral lymphomas.

Perinatal induction of medulloblastomas in Syrian golden hamsters by a human polyoma virus (JC).

Medulloblastomas, originating from cells of the internal granular layer of the cerebellum, developed in Syrian golden hamsters 3--6 months after inoculation at birth intracerebrally and sc with JC virus, a papovavirus isolated originally from a human case of the demyelinating disease progressive multifocal leukoencephalopathy. The lesion is compared with the human medulloblastoma of childhood. JC virus is the first infectious agent to produce such a neoplasm in any species.

Progressive multifocal leukoencephalopathy. A cause of visual loss.

A patient had been treated for chronic lymphocytic leukemia for five years before developing visual blurring as the manifestation of occipital lobe lesions of progressive multifocal leukoencephalopathy (PML). The disease is caused by an infection of the CNS oligodendrocytes by a papovavirus, whose replication is facilitated by an impairment of the host's cell-mediated immunologic system. The multiplying virus destroys the oligodendrocytes and causes extensive demyelination of the white matter of the brain. Antiviral agents, such as cytarabine, may be beneficial in treating patients with PML.

Differential neurooncogenicity of strains of JC virus, a human polyoma virus, in newborn Syrian hamsters.

The neurooncogenicity recently isolated strains of the human polyoma virus, JC virus, was determined by intracerebral inoculation of newborn Syrian golden hamsters. All three strains produced malignant brain tumors in a majority of inoculated animals during a 6.5-month observation period. The results obtained with the MAD-2 strain, 19 of 20 animals with cerebellar medulloblastomas and 0 of 20 animals with pineal gland tumors, were quite similar to those observed previously with the prototypic strain of JC virus, MAD-1. Inoculation of the MAD-4 strain, however, resulted in 10 of 22 animals with pineal gland tumors and only 10 of 22 animals with tumors in the cerebellum. The MAD-3 strain was neurooncogenic, but too few animals lived to be weaned to provide significant additional information. The basis for the apparent predilection of the MAD-4 strain for the pineal gland is unknown. Two hamsters in the experiment developed extracranial neuroblastomas.

Modification of hydroxyindole-O-methyltransferase activity in experimental pineocytomas induced in hamsters by a human papovavirus (JC).

Hydroxyindole-O-methyltransferase (HIOMT) (HIOMT) activity was studied and compared in 7 pineal tumors (pineocytomas) induced in vivo in Syrian hamsters after postnatal inoculation with strains of human papovavirus (JC). Levels of tumor HIOMT activity were correlated with degree of cytologic differentiation of the pineocytes of each neoplasm, as judged from electrom micrographs. The relative capacities of the HIOMT in the tumors to methylate three related substrates gave similar ratios in the individual tumors and were not different from those of HIOMT in normal hamster pineal glands.

Experimental pineocytoma of the Syrian hamster induced by a human papovavirus (JC). A light and electron microscopic study.

The gross, light and electron microscopic features of a pineocytoma of the Syrian hamster (mesocricetus auratus) which had been induced by a human oncogenic virus (JC papovavirus) have been defined. For comparison, adult hamster pineal tissue was studied, and the literature was consulted in regard to other pineal parenchymal tumors, and to pineal cell differentiation during ontogeny and phylogeny. Many differentiated tumor cells contained organelles, such as vesicle-crowned lamellae (synaptic ribbons) and microtubular sheaves, as consistent with adult hamster pineocytes. Some cells showed rudimentary photoreceptor-like differentiation as consistent with fetal hamster pineocytes and with cells seen in the pineal systems of some lower vertebrate species. Such tumor cells had lumen-directed specialized cytoplasmic extensions which, by their richness in mitochondria and presence of centrioles and striated rootlets, resembled inner segments. Extending 9+0 cilia were accompanied by occasional lamellar whorls. Oncogenesis seems to have simulated different stages of hamster pineal ontogeny. This observation would support the theory that the secretory mammalian pineocyte derived phylogenetically from the true photoreceptor cell of the pineal system of fishes and amphibians. The possible influences of host and of virus in the accomplishment of tumor morphology were discussed. This tumor differed considerably in pattern and cell detail from the only other pineocytoma studied previously by electron microscopy. It is the first experimentally induced pineocytoma.