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Biomedical subjects

D Santoli

Publications and source records attributed to D Santoli.

At least 91 records · Page 5Linked to original sources

Cell-mediated cytotoxicity against virus-infected target cells in humans. I. Characterization of the effector lymphocyte.

In a 51Chromium release assay performed without immune sera, purified lymphocytes from randomly selected healthy donors killed significantly more allogeneic virus-infected than noninfected fibroblasts. An incubation period of 18 to 20 hr was optimal for determining cytotoxicity, but increased cytotoxic activity against infected cells was observed as early as 6 hr from the start of the test. No correlation could be found between donors' antibody titers to any virus tested and the cytotoxic efficiency of their effector cells: virus-infected cells were lysed by both seropositive and seronegative individuals. With several different cell fractionation techniques, the effector cell in this system could not be distinguished from the human natural killer cell, which is a lymphocyte with receptors for the Fc fragment of IgG but with no surface immunoglobulin. When lymphocytes were separated on the basis of ability to form rosettes with neuraminidase or AET (2-aminoethylisothiouronium bromide hydrobromide)-treated sheep erythrocytes, the majority of cytotoxic activity was consistently recovered in the nonrosetting fraction. A portion of it, however, was always present in the rosetting fraction and was, again, mediated by lymphocytes carrying receptors for the Fc fragment of IgG.

Adult↗

Spontaneous cell-mediated cytotoxicity in humans. Distribution and characterization of the effector cell.

When lymphocytes from healthy donors were tested as effector cells, the cytotoxic activities observed in spontaneous and in antibody-dependent cell-mediated cytotoxicity were positively correlated. However, with lymphocyte preparations obtained from renal patients, a dissociation between the two activities was occasionally observed. Human natural killer cells are lymphocytes, with receptors for the Fc fragment of IgG molecules, but with no surface immunoglobulin. Their cytotoxicity is reduced by the presence of granulocytes or monocytes. After separation of rosetting and non-rosetting cells with AET- (2-aminoethylisothiouronium bromide hydrobromide) or neuraminidase-treated sheep erythrocytes, the majority of the activity was recovered in the non-rosetting fraction, but a portion of it was present consistently in the rosetting cell fraction. Cells in the latter fraction also displayed receptors for the Fc fragment of immunoglobulin G.

Adult↗

Acute optic neuritis: a virological study in relation to multiple sclerosis.

Previous ultrastructural examination of peripheral blood lymphocytes revealed the presence of intranuclear filamentous structures in multiple sclerosis (MS) and in some optic neuritis (ON) patients. The present investigation was undertaken in the attempt to correlate the presence of such structures with the etiology of ON and MS and possibly to demonstrate the viral origin of the filaments. Suitable virological and serological techniques were used to detect and isolate infectious agents from peripheral blood samples and body excretions of 12 monosymptomatic ON patients at their first acute attack. Nevertheless, any efforts to demonstrate the presence of a virus in these patients have been unsuccessful: no evidence of active viral infection was obtained by serological studies of serum and cerebrospinal fluid samples, nor could viral antigens or inclusions be observed by immunofluorescence and cytochemical analysis. Negative results were also obtained from studies performed in parallel on MS patients and various controls. The significance of the failure to isolate infectious agents from either ON and MS patients is discussed.

Acute Disease↗

Evaluation of the effect of ammonium chloride treatment on the activity of human effector cells in antibody-dependent and spontaneous cell-mediated cytotoxicity.

The effect of ammonium chloride treatment, used to eliminate contaminating red blood cells from lymphoid cell preparations, on the cytotoxic activity of human effector cells in antibody-dependent and spontaneous cell-mediated cytotoxicity was quantitatively determined. No significant difference between the cytotoxic efficiency of treated and untreated cell preparations was observed.

Ammonium Chloride↗

Persistent parainfluenza type 1 (6/94) infection of brain cells in tissue culture.

A state of persistent infection with parainfluenza type 1 virus (6/94 strain) was established in cultures of human and bovine brain cells. Following primary infection of human brain cells, viral cytopathic effect (CPE) and hemadsorption (HAD) depended on the multiplicity of infection. After persistent infection was established the virus rapidly became cell-associated; no CPE occurred and no viral antigen was detectable by HAD, immunofluorescence (FA), or immunoprecipitation. Infectious virus could be recovered only by fusion or cocultivation. This was in marked contrast with infected bovine brain cells, where, following primary infection, little or no CPE occurred. A productive infection rapidly evolved and persisted without CPE, but with 100 per cent HAD and FA positive cells.

Animals↗

Unusual intranuclear filaments in the circulating lymphocytes of patients with multiple sclerosis and optic neuritis.

Mononuclear cells obtained from the peripheral blood of patients with multiple sclerosis (MS) and optic neuritis (ON) were examined by electron microscopy. Unusual intranuclear filaments identical with the so-called paramyxovirus-like filaments were found in circulating lymphocytes from 5 patients with MS and 3 with ON. They were observed in 0.1 to 0.5% of the mononuclear cells. Electron microscopy of the blood cells from 5 other patients with ON, 3 patients with neurologic diseases other than MS, and 4 normal subjects failed to reveal similar structures. The filaments were also absent from blood cells from blood cells from normal subjects during the process of autolysis.

Adolescent↗

HLA-related control of spontaneous and antibody-dependent cell-mediated cytotoxic activity in humans.

Normal human lymphocyte preparations were tested for their ability to lyse both antibody-coated and unsensitized human target cell lines. The capacity to induce these two types of cell-mediated cytotoxicity, antibody-dependent (Ab-CMC) and spontaneous (Sp-CMC), was detected by a 51Cr release assay. The reactivity of lymphocytes from 1. individual donors showed a positive and highly significanlty correlation between Ab-CMC and Sp-CMC, leading to the hypothesis that the same type of effector cell is involved in the two cytotoxic mechanisms. Lymphocytes from male donors were about twice as effective as those from female donors in both systems. Moreover, the effector cells from male donors carrying HLA antigens A3 and B7 displayed a significantly lower reactivity in both Sp-CMC and Ab-CMC when compared with lymphocytes from male donors bearing any other HLA haplotype. The possible significance of the hyporeactivity of lymphocytes from normal subjects with HLA-A3,B7 haplotype in relation to an increased susceptibility to multiple sclerosis (MS) is discussed.

Adult↗

Human brain in tissue culture. I. Acquisition, initial processing, and establishment of brain cell cultures.

This paper details the in vitro techniques used to establish cells in culture from the brains of 40 patients, most of whom had chronic neurologic disease. The clinical and pathologic features of these patients are given. The success in establihsing cell lines was dependent upon the origin of tissue (biopsy vs. autopsy), the site of removal from the brain, and various environmental and technical manipulations in vitro.

Adolescent↗

Human brain in tissue culture. II. Studies of long-term cultures.

This paper describes the techniques used to maintain and reconstitute from storage adult human brain cells in culture. Growth characteristics, cell morphology, lifespan, and karyotypic analysis of cell lines derived from patients with multiple sclerosis (MS), non-MS neurologic diseases, and normal brains are compared.

Aging↗

Human brain in tissue culture. III. PML-SV40-induced transformation of brain cells and establishment of permanent lines.

Cells from explants and monolayer subcultures of adult human brain obtained from biopsies or autopsies of ten multiple sclerosis (MS) cases, one case each of Jakob-Creutzfeld (JC) and amyotrophic lateral sclerosis (ALS) and three cases with no central nervous system (CNS) involvement were transformed with PML-SV40 virus. Transformation was effected to establish permanent lines of these particular adult brain cells so that sufficient quantities would be available for other research projects. The procedure previously used to transform human fibroblasts (Koprowski et al., '62) was successfully applied to human brain cells. The success of transformation was dependent on the growth condition of the cultures at the time of infection. Events occurring after viral infection and during the pre-transformation and the post-transformation phases are described.

Amyotrophic Lateral Sclerosis↗

Human brain in tissue culture. IV. Morphological characteristics.

The morphology of cells from normal and diseased brains and PML-SV40 transformed brain cells grown on glass cover slips in cultures is described. Seven types of cells are identifiable, four of which are probably of mesenchymal origin whereas the remaining three are neurologlial. A classification on the basis of the morphologic features is presented.

Amyotrophic Lateral Sclerosis↗

Hydrocephalus produced by the 6/94 virus; A parainfluenza type 1 isolate from multiple sclerosis brain tissue.

The 6/94 virus, parainfluenza type 1 isolate from multiple sclerosis brain tissue, produced hydrocephalus in newborn Syrian hamsters. All animals developed clinical disease and died within a week. Ependymal cells lining the aqueduct of Sylvius became necrotic and fused, resulting in obstructive hydrocephalus. The 6/94 virus antigen was seen in ependyma and meninges. Paramyxovirus nucleocapsids were seen within cytoplasm of ependymal cells. Virus was recovered from hamster brains for only two days. Infectious virus could be recovered from brains grown in vitro as explants for 21 days. No evidence of rising hemagglutination-inhibiting antibody was noted for up to one month after infection. Intraperitoneal or subcutaneous injection of 6/94 virus did not produce hydrocephalus. HA2 virus and the temperature sensitive mutant of HA2 virus failed to produce hydrocephalus, while Sendai virus caused lesions similar to those of 6/94 virus.

Animals↗

Establishment of continuous multiple sclerosis brain cultures after transformation with PML-SV40 virus.

PML-SV40 virus was used to transform cells from explants and monolayer subcultures of human brain derived from biopsies or autopsies of MS cases, as well as from 1 case of Jakob-Creutzfeldt disease (JC), 1 case of amyotrophic lateral sclerosis (ALS) and several cases without CNS disease. The success of transformation depended on the growth condition of the culture at the time of infection but at present at least 14 cell lines, 10 derived from MS, 2 from JC, 1 from ALS and 1 from a normal non-CNS patient are maintained in continuous culture.

Amyotrophic Lateral Sclerosis↗