Studies on the development of a live attenuated mumps virus vaccine. I. Attenuation of the Hoshino "wild" strain of mumps virus.
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Animals↗
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Influence of preincubation of primary dog kidney cell cultures on the multiplication of attenuated mumps virus.
An attenuated mumps virus strain multiplied in primary dog kidney cells to significantly higher titres in cultures preincubated for 1-5 weeks at 20-25 degrees C before inoculation as compared with cultures infected immediately after their growth became confluent. The nature of this phenomenon is discussed.
Animals↗
Virulence and persistence of three prototype strains of mumps virus in newborn hamsters.
Neuroadapted mumps virus (NMV) produces widespread central nervous system (CNS) disease and death after intracerebral (i.c.) inoculation of newborn hamsters. After intraperitoneal (i.p.) inoculation, NMV causes disseminated disease, moderate mortality and it persists in CNS tissues. Low tissue culture passage isolates of wild mumps virus do not establish CNS infection after i.p. inoculation; after i.c. inoculation they cause limited though persistent infection with little acute mortality. The biological behavior of a highly passaged vaccine strain of mumps virus (Jeryl-Lynn) is more similar to NMV than to the wild strain in its behavior in the newborn hamster.
Adaptation, Biological↗
Immunologic significance of the mumps virus skin test in infants, children and adults.
The biologic validity of cell-mediated immunity to mumps virus was evaluated in 395 children, adolescents and adults. The study protocol included the determination of cutaneous delayed hypersensitivity to viral and avian control antigens and in 79% of the subjects an essential double bleeding was performed before and after mumps virus skin test for assay of neutralizing antibody. Seven per cent of subjects expressed sufficient delayed hypersentitivity to the control antigen to erase an apparently positive mumps virus skin test. Anamnestic conversions from seronegativity to seropositivity, elicited by the mumps virus skin test, increased from 4% in children to 25% in adults, which suggests waning B-cell recognition of prior mumps virus infection in adults. Although pregnancy diminished the difference (p smaller than .001), adults showed greater cutaneous delayed hypersensitivity to mumps virus antigen than did children (p smaller than .001), suggesting that mumps virus reinfection or persistence induced the escalation of more sensitive T-cell recognition with increasing age. Humoral immunity, assessed by the double bleeding technique in the vast majority of individuals, rose form 16% (1-4 years), 45% (5-9 years) and 80% (10-14 years) to 94% in adolescents and adults. Ordinarily 75-95% in other age groups, the decline of correlation between mumps virus cellular and humoral immunity to 60% in school children may result from prior parainfluenza virus infection, inconsistent potency of the skin test antigen, concurrent immunosuppressive infection, and lagging induction of mumps virus cellular immunity in recently infected individuals. Immunologic study of a large colony of subhuman primates failed to establish an hierarchial antigenic interrelationship among mumps virus and two additional paramyxoviruses.
Adolescent↗
Attenuated mumps virus therapy of carcinoma of the maxillary sinus.
This paper presents treatment of carcinoma of the maxillary sinus with an attenuated mumps virus. The mumps virus was inoculated into two cases of very advanced carcinoma of the maxillary sinus by local and general injection. Shortly after injection, relief from severe cancer pain and growth inhibition or necrosis of tumor were observed in both cases. In this communication, we discuss a possible mechanism of antitumor effect of mumps virus by utilizing immunologic tests and interferon assay.
Carcinoma, Squamous Cell↗
Morphological differences in the central nervous system and organs of monkeys inoculated intracerebrally with virulent and attenuated strains of mumps virus.
The degree of attenuation of mumps virus was studied in intracerebrally inoculated Macaca mulatta monkeys. The virulent Leningrad-3 strain and its attenuated variant as well as the attenuated Sophia-6 strain were used. The nature of pathological alterations in the neurons pointed to neurotropism of only the virulent strain. Morphological changes caused by attenuated and virulent strains in the brain structures, in the epithelium and glandular structures of the salivary glands differed considerably and may be used as a kind of markers of neurovirulence and the degree of attenuation of mumps virus strains.
Animals↗
Induction of congenital hydrocephalus with mumps virus in rhesus monkeys.
Rhesus monkey fetuses were inoculated intracerebrally with wild-type mumps virus near the beginning of the last third of the gestation period. Within three days after inoculation, mumps virus was isolated from many fetal tissues. Thirteen animals receiving virus were delivered at term. Five of these showed slight to severe hydrocephalus. Hydrocephalus was most prominent in the posterior horns of the lateral ventricles, but other lesions occurred at various levels of the ventricular system. Virus was isolated from three animals at one day of age (two months after inoculation). However, mumps virus was not recovered from one-month-old monkeys. The recovery of mumps virus from newborn rhesus monkeys two months after inoculation suggests that attempts should be made to document similar persistence of the virus in humans. Furthermore, this model indicates that mumps virus infection in humans may result in hydrocephalus.
Amniotic Fluid↗
Structural polypeptides of mumps virus.
The structural polypeptides of egg grown mumps virus were analysed by SDS-polyacrylamide-slab-gel electrophoresis. Mumps virions contained eight major polypeptides with mol. wt. of 75, 73, 71, 61, 47, 44, 42 and 40 X 10(3). The 75 K and 61 K polypeptides were glycosylated. In virions treated with pronase and trypsin, the 75 K glycoprotein was removed more readily from the virus than the 61 K glycoprotein. The gradual removal of the 75 K glycoprotein was paralleled by a decrease of haemagglutinating activity. The large glycoprotein was cleaved into a 40 K glycoprotein by trypsin treatment. Pronase and trypsin treatment also removed the smallest 40 K non-glycosylated polypeptide. Thus this polypeptide appears to be located on the outside of the virion and probably represents a cleavage product of the large glycoprotein. Treatment of virions with 2% Triton-X 100 under alkaline conditions in the absence or presence of 2 M-KCl solubilized the two glycoproteins and a fraction of the 71 and 44 K polypeptides, but not the 73 and 47 K polypeptides. The two smallest polypeptides were solubilized by treatment with 2% Triton X-100 in the presence of 2 M-KCl. Since the 40 K polypeptide was interpreted to represent a cleavage product of the large surface glycoprotein the 42 K polypeptide was proposed to represent the membrane protein of mumps virus. The 44 K polypeptide co-migrated with Vero cell actin. The nature of the 47 K polypeptide could not be determined, but it is probably located in the central part of the virus. The 73 K polypeptide and in some experiments also the 71 K polypeptide were found in purified nucleocapsid preparations. It is concluded that mumps virus has a general polypeptide composition similar to other paramyxoviruses. However, the molecular weights of the different polypeptides of mumps virus differ markedly from the corresponding polypeptides in Newcastle disease virus and Sendai virus.
Glycoproteins↗
Polypeptide composition of mumps virus.
The Enders strain of mumps virus grown in ovo was purified by differential and equilibrium sucrose gradient sedimentation. Purified virus contained seven polypeptides of mol. wts 68,000, 66,000, 61,000, 54,000, 52,000, 49,000, 47,000. Nucleocapsids isolated from DOC-treated virus contained two polypeptides of mol. wts 66,000 and 61,000.
Molecular Weight↗
Rapid diagnosis of mumps virus infections by immunofluorescence methods.
Mumps and its complications, particularly meningoencephalitis, is an important disease problem, and more rapid diagnostic methods are desirable. A study was made of immunofluorescence methods for the early detection of mumps virus isolated in cell cultures, or adsorbed directly from clinical specimens onto guinea pig erythrocytes. A specific diagnosis could be made in hours to 2 or 3 days utilizing immunofluorescence methods, in contrast to about 6 days by standard methods. Details of the direct immunofluorescence methods are presented, to encourage wider application in clinical virology laboratories.
Animals↗
A persistent infection of baby hamster kidney-21 cells with mumps virus and the role of temperature-sensitive variants.
A persistent infection of baby hamster kidney-21 (BHK-21) cells with mumps virus (BHKpi) was maintained for over 60 cell passages in the absence of antiserum. Viral persistence was demonstrated in the cultures by hemadsorption, immunofluorescence, multinucleate syncytia, and released mumps virus at the level of 10(2)--10(3) fluorescent focus-forming units/ml. No detectable levels of interferon were found in cultures persistently infected with mumps virus. Approximately 85--95% of the cells contained viral antigens. Nuclear fluorescence was observed in the persistently infected cells. Mumps virus from persistently infected clutures (MuVpi) was more heat-labile than wild-type mumps (MuVo) when subjected to 40 degrees C. BHKpi cells had a more rapid doubling time and a higher cloning efficiency in soft agar in comparison to BHK-21 cells. MuVpi was also found to be temperature-sensitive. The temperature-sensitivity of MuVpi was determined by the efficiency of plating at 33 degrees and 39 degrees C. MuVpi readily established a persistent infection in BHK-21 cells with less cytopathology than MuVo, and released temperature-sensitive virus.
Animals↗
[Comparative sensitivity of 4 different cell cultures in the isolation and identification of mumps virus].
Three different continuous cell lines (Am 57, Hela, Vero) and primary cell culture of human embryo kidney are compared with regard to their susceptibility for isolation of mumps virus from saliva and cerebrospinal fluid. For this purpose the Vero cell line appeared to be more suitable among the studied cell systems regarding the following reasons: it allows the highest percentage of mumps virus isolation. The cytopathic effect caused by mumps virus occurs comparatively rapidly in this cell culture. This cytopathic effect is typical enough to allow for a preliminary diagnosis of mumps virus.
Animals↗
Cell- and antibody-mediated responses to measles and mumps viruses in experimental animals.
Monkeys and guinea pigs were immunized with various doses of measles and mumps viruses. A cell-mediated response measured by leukocyte migration inhibition occured during the first two weeks after immunization. In case of measles virus, marked migration inhibition occurred already on the 7th day in monkeys and guinea pigs immunized with high virus doses. The response declined as early as 14 days after immunization in contrast to the group of guinea pigs inoculated with a lower virus dose. In these animals the response sustained for up to 50 days. Immunization of animals with various doses of mumps virus was similar and positive responses were obtained in the absence of detectable antibodies. Even 2.5 TCID50 of mumps virus induced the response although more irregularly than 250 or 2500 TCID50 virus.
Animals↗
Effects of ribavirin on BHK-21 cells acutely or persistently infected with mumps virus.
The effects of ribavirin on BHK-21 cells acutely infected with mumps virus were compared to the effects of the drug on the same cell line persistently infected with mumps virus. Visible cytotoxicity was minimal for both cell types; however, there was an inhibition of cell replication with increasing drug concentrations. Ribavirin had marked antiviral activity against both the acute and persistent infections as determined by an inhibition of hemadsorption plaque formation, decreased immunofluorescence, and a reduction in the release of infectious virus. Even after the drug had been on the persistently infected cells for 72 h, there was still antigen production detectable by immunofluorescence, although the cells no longer hemadsorbed chicken erythrocytes. Ribavirin removal from both types of infection resulted in the renewed synthesis of virus.
Animals↗
Detection of mumps virus antigens in Hodgkin's disease tissues.
Mumps virus antigens were demonstrated in biopsied tissues from Hodgkin's disease patients by indirect immunofluorescence. Impression smears from ten lymph node and two spleen specimens revealed viral antigens in the nucleus, cytoplasm or both. Measles virus antigens were detected in six out of seven Hodgkin's disease tissue (lymph nodes) both in the nucleus and cytoplasm. All tissues tested for the presence of Newcastle disease virus (NDV, an avian paramyxovirus) antigens were negative. Control tissues were obtained from patients with non-Hodgkin's lymphomas, breast cancers, adenocarcinomas and a number of other disease processes. In control tissues mumps antigens were detected in seven out of 31 specimens and measles antigens in nine out of 18 tissues.
Adolescent↗
Studies on the use of mumps virus for treatment of human cancer.
Purified mumps virus (Urabe strain) was given mainly by intravenous injection to a total of 200 patients with cancer. The only adverse clinical reaction was transient mild fever in about half the patients. The beneficial clinical effects were as follows; decrease or disappearance of ascites and edema of the lower limbs at high rates (26/37 and 4/4, respectively), usually within a week after treatment: decrease or stoppage of cancerous bleeding in 30 of 35 patients: decrease or disappearance of pain in most of the patients: and tumor regression in 26 patients with cancer of the breast, rectum, ileocaecum, thyroid gland, uterus, skin, etc. Histologically, the virus-treatment caused shrinkage of nuclei and vacuolization of the cytoplasma of tumor cells, but the degenerative changes of tumor cells were not so great as those after chemotherapy or radiotherapy. Infiltration of lymphocytes, fibrosis and collagenesis occurred around tumor tissues, where necrosis or exfoliation of tumor cells was frequently observed.
Adenocarcinoma↗
Differences in antibodies to the surface components of mumps virus after immunization with formalin-inactivated and live vaccines.
Military recruits with or without antibodies to mumps virus were immunized with live or formalin-inactivated mumps virus vaccines. Antibodies to the two major surface components of the virus, the hemagglutinin and the hemolysin, were identified separately. Immunization with the live vaccine did not change the humoral immunity in individuals with detectable antibodies. In seronegative recruits immunization with the live vaccine induced an antibody response against both the hemagglutinin and the hemolysin corresponding to the quality of the immune response after natural infection. In contrast, the formalin-inactivated vaccine only induced an antibody response against the hemagglutinin. This effect was found after immunization of both seronegative and seropositive individuals. The latter displayed a selective boosting of titers of hemagglutination-inhibiting antibody. These findings may have relevance for the interpretation of the short-lived immunity after the use of formalin-inactivated mumps vaccine without repeated boosting.
Antibodies, Viral↗
[Biological properties of the mumps virus--behavior in the rct marker].
12 different mumps virus strains or their variations were studied in the rct-marker in dog kidney cell cultures at 32 degree and 39 degree C. The results obtained were compared with those of the T50 marker ascertained earlier revealing considerably coincident data. Changes in culture conditions became clearly evident in both markers. Relations between their behaviour in vitro in vivo are discussed.
Animals↗