STUDIES OF THE HEMOLYSIS OF RED BLOOD CELLS BY MUMPS VIRUS : II. THE RELATIONSHIPS OF HEMAGGLUTINATION, VIRUS ELUTION, AND HEMOLYSIS

The relationship between hemagglutination and hemolysis by the mumps virus has been studied under conditions which affect (a) the receptors of chicken red cells and (b) the adsorption and subsequent elution of the virus from these cells. The results show that the hemolytic action of the virus appears to involve some of the same receptor areas of erythrocytes that are implicated in hemagglutination. Materials such as allantoic fluid, egg white, and red cell extract, which inhibit the agglutination of chicken red cells by mumps virus, also interfere with its hemolytic activity. Of these inhibitors, egg white and red cell extract, which are readily destroyed by the virus during incubation at 37°C., exert a greater antagonistic effect on hemagglutination than on hemolysis. Heated mumps virus or unheated influenza virus interferes with the hemolysis of red cells by untreated mumps virus. Though hemolysis takes place during elution of the virus after its adsorption on the red cell, the processes are apparently distinct. The hemolytic activity is easily affected by certain conditions of pH and temperature which have no effect on the ability of mumps virus to adsorb on and elute from red cells.     

IMMUNITY IN MUMPS : I. EXPERIMENTS WITH MONKEYS (MACACUS MULATTA). THE DEVELOPMENT OF COMPLEMENT-FIXING ANTIBODY FOLLOWING INFECTION AND EXPERIMENTS ON IMMUNIZATION BY MEANS OF INACTIVATED VIRUS AND CONVALESCENT HUMAN SERUM

Since the experimental data have already been discussed at some length, it is here necessary only to review those observations which appear to be of most significance. The manifestations of the disease we have produced in monkeys have in general been similar to those described by previous workers. It has been found that members of the species M. mulatta are regularly susceptible to infection with the virus of mumps, provided they have not been in contact with animals which have recently been infected. Such contact may be followed by the development of resistance to inoculation unattended by overt signs of infection. Following the intraparotid injection of virus into a susceptible animal, an antigen appears in the gland which reacts specifically in the complement fixation test with sera of monkeys which have recovered from an attack of mumps. This antigen has not been demonstrated in any other organ of the infected monkey which has been examined, nor does it persist or increase in amount when introduced into the brain or the testes. Attempts to propagate it in the tissues of other animal species including the developing hen''s egg and tissue cultures have so far been uniformly unsuccessful. The antigen is relatively thermostabile, resisting a temperature of 65°C. for at least 20 minutes at the optimal hydrogen ion concentration. It is unimpaired by a concentration of formalin of 0.3 per cent. Together with the virus it may be preserved indefinitely in the frozen state. The maximal quantity of antigen in the gland, which may be considerable, is attained on or about the 5th day following inoculation of the virus. An antibody which reacts with the antigen appears in the serum from 8 to 14 days after infection. This antibody has not been found in the serum of normal monkeys which have had no opportunity for contact with the virus. It has been demonstrated during convalescence in the serum of all animals which have been examined. The antibody which is present in large amounts soon after recovery, although decreasing in concentration with the lapse of time, may continue to be present for many months. At 62°C. for 20 minutes its activity is not reduced and on storage at 4°C. its titer has remained unaltered for over 2 years. A similar factor which appears in human beings convalescent from mumps is less stabile, since its activity is reduced appreciably under these conditions. In the CO₂ cabinet, however, both antibodies can be preserved for long periods of time. The definition of these reagents has afforded means whereby the simian infection with the virus of mumps, whether it is actively in progress or has occurred at some time in the past, can be recognized. Upon the establishment of these facts, the estimation of the quantity of antigen in the glands of vaccinated animals was employed as an index of their resistance in studies on the effect of the injection of formolized and alumprecipitated formolized suspensions of infected parotid gland. It has been shown that such vaccines, in which the virus has been rendered inactive, lead to the production of complement-fixing antibody. In about 60 per cent of the vaccinated animals, moreover, evidence of increased resistance was obtained as indicated by the partial or complete suppression of the formation of antigen in the inoculated gland. In certain instances no evidence of resistance was observed, yet antibody was found to be present. It is clear, therefore, that resistance is not determined solely by the presence of the complement-fixing antibody. The possibility, however, of its participation as a factor in immunity cannot be excluded. Employing the same criterion of resistance, the virus-neutralizing capacity of a pool of human mumps convalescent sera and of a globulin concentrate prepared from the pool has been studied. Neither of these materials was found to prevent entirely the formation of antigen, although each of them appeared to exert some inhibitory effect.     

陕西省铜川市流行性腮腺炎流行特征分析

目的 了解陕西省铜川市流行性腮腺炎流行特征,为有效控制该病提供参考依据。 方法 采用描述流行病学方法,对铜川市2004-2009年流行性腮腺炎病例进行统计分析,采集腮腺炎患者唾液标本分离病毒,并在铜川市随机抽取健康人群采集静脉血检测腮腺炎IgG抗体。 结果 铜川市2004-2009年共报告腮腺炎病例1155例,年均发病率为22.87/10万,发病呈现两个季节高峰,以5~15岁儿童发病为主,职业分布上以中、小学生为多;共采集健康人群血样452份,腮腺炎抗体阳性率74.78%,抗体几何平均浓度(GMC)为830.51 U/ml;腮腺炎抗体阳性率及GMC均先随年龄增长而增高,抗体阳性率7~10岁最高为100%,GMC 5~6岁最高为1923.14 U/ml,以后随年龄增加抗体阳性率及GMC均略有下降,但仍然维持在较高水平。结论 近年来铜川市腮腺炎发病一直维持在较高水平,铜川市腮腺炎流行是由腮腺炎病毒F基因型引起,今后流行性腮腺炎防控的重点人群是7岁以下的儿童,尤其是1~3岁儿童。     

Secretory and serum immunoglobulin class-specific antibodies to mumps virus after a natural mumps infection

Immune responses in serum and saliva were studied in 50 individuals following a natural exposure to mumps virus, using ELISA for detection of mumps-specific IgG, IgM and IgA antibodies.Laboratory diagnosis of acute mumps using a single saliva sample offers a convenient alternative to collection of blood: high levels of mumps IgM were found in all serum and saliva samples taken from patients with acute mumps. Furthermore, in such patients, levels of mumps IgA were considerably higher in saliva compared with serum, with the exception of three patients. Conversely, mumps-specific IgM and IgA were present in neither serum nor saliva samples of patients with a remote mumps infection.Saliva samples are not suitable for immune status (IgG) determinations because high levels of mumps IgG antibodies were present only in serum samples of patients with either a recently acquired (≥ 15 days after onset) or a remote mumps infection.Serum and saliva mumps-specific IgA antibodies appeared to be a reliable acute phase marker as, like IgM antibodies, they were present in all tested serum/saliva samples taken shortly after the onset of the disease (1–5 days), were detected for a limited period of time (up to 3 months), and were absent from serum and saliva samples of patients with a remote infection.     

Studies of delayed hypersensitivity in vitro. II. Delayed hypersensitivity in experimental mumps virus infections

Guinea pigs experimentally infected with mumps virus develop a delayed, hypersensitive skin reaction following the intradermal injection of heat-inactivated mumps virus. This in vivo hypersensitivity is accompanied by a state of cellular hypersensitivity which can be demonstrated in vitro by the addition of mumps viral antigen to cultures of splenic macrophages, following which they become less motile and undergo lysis. These observations support the hypothesis that the state of hypersensitivity which develops early in mumps virus infections may have a role in the pathogenesis of the disease.     

A FURTHER STUDY OF EXPERIMENTAL PAROTITIS

A new series of inoculations into cats of the filtered sterile salivary secretions derived from cases of parotitis has been described. They confirm the observations made in 1915–1916 and extend them to include the epidemic parotitis occurring in our military forces. Incidentally confirmatory evidence of the filterable nature of the causative agent of mumps has been obtained. It has been determined that the saliva of man and of inoculated cats, and the inoculated glands of the latter animals, contain the filterable, infective agent. The lesions present in the inoculated organs conform to those described in our first publication. In addition, the lymph glands adjacent to the salivary glands on the uninoculated side were sometimes found to be swollen and to exhibit microscopic lesions. Probably the involvement resulted from salivary and lymphatic infection. The "virus" of parotitis was detected most readily in the saliva during the first 3 days of the disease, less easily on the 6th day, and not at all on the 9th day. It was detected also in the blood of patients showing marked constitutional symptoms, and in the saliva of a case of recurrent mumps at the periods of enlargement of the parotid glands, but not 2 weeks after the swelling had subsided. It was not detected in the cerebrospinal fluid.     

IMMUNITY IN MUMPS : VI. EXPERIMENTS ON THE VACCINATION OF HUMAN BEINGS WITH FORMOLIZED MUMPS VIRUS

The results observed after experimental inoculation of active mumps virus into 41 vaccinated and 32 unvaccinated children,—with the consent of their parents or guardians,—indicated that formol-inactivated mumps virus obtained from the parotid gland of the infected monkey and employed as a vaccine in the manner which has been described increased the resistance of about half of those to whom it was administered.     

Mumps: An Emergency Medicine-Focused Update

Background

Mumps is a Paramyxoviridae virus. This disease was rampant prior to introduction of the measles, mumps, and rubella vaccine, resulting in decreased incidence. This disease has demonstrated several outbreaks.

ST. LOUIS ENCEPHALITIS : TRANSMISSION OF VIRUS TO CHICKENS BY INFECTED MITES DERMANYSSUS GALLINAE AND RESULTING VIREMIA AS SOURCE OF VIRUS FOR INFECTION OF MITES

Transmission of the virus of St. Louis encephalitis to normal chickens by the bite of infected mites (Dermanyssus gallinae) has been demonstrated. Both experimentally infected and naturally infected mites were shown to be capable of transferring the virus of St. Louis encephalitis to chickens by bite. Virus is present in the blood of such chickens in small amounts, so that demonstration of viremia was possible only by utilizing chorioallantoic passage in hens'' eggs. However, there is sufficient virus present in the blood for uninfected chicken mites to acquire the virus by feeding on chickens in which viremia has resulted from previous bite of infected mites. Thus it has been shown that the arachnid vector Dermanyssus gallinae is capable of transmitting the virus of St. Louis encephalitis to normal chickens by bite and that such chickens can serve as a source of virus for uninfected mites.     

Isolation of mumps virus from human beings with induced apparent or inapparent infections

Exposure of fifteen children to mumps virus of fifth amniotic passage in chick embryos led to involvement of the salivary glands in six, orchitis in the absence of other manifestations of mumps in one, and to no signs of illness in eight. Attempts to isolate virus from the saliva of these individuals gave the following results: 1. All patients with involvement of the salivary glands excreted virus beginning on the 11th to 15th day after exposure, 2 to 6 days prior to onset of clinical signs of disease and extending up to the 4th day of illness. 2. The patient with primary orchitis without any recognized involvement of the salivary glands excreted virus for 2 days, beginning on the 15th day after exposure and 10 days prior to his illness. 3. Six of the eight children classified as having inapparent infections because of their serologic response in the absence of clinical signs of illness, began to excrete virus on the 15th to 16th day after exposure for from 1 to 9 or more days. The epidemiologic significance of these data is discussed.     

A CARRIER STATE OF MUMPS VIRUS IN HUMAN CONJUNCTIVA CELLS : I. GENERAL CHARACTERISTICS

Mumps virus produced a carrier state in human conjunctiva cells that was maintained for more than 100 subcultures over a period of 3 years. Antiserum in the medium was not required. The virus had little apparent effect on the cells which grew at a rate similar to uninfected control cells. Mumps virus was regularly found in the culture medium at levels about 0.9 log higher than the cell-associated virus. When first tested after 30 subcultures, the virus was found to have lost its cytopathogenicity for cells ordinarily susceptible to mumps virus, but was identifiable as mumps virus by neutralization with specific antiserum. Use of fluorescein-labeled antiserum revealed that 80 to 95 per cent of cells in the carrier cultures contained mumps virus antigen. The antigen was concentrated in a few sharply circumscribed, discrete masses in the cell cytoplasm rather than in many granules throughout the cytoplasm as is characteristic of cell infection by cytopathogenic mumps virus. The carrier cultures were resistant to the destructive effect of a cytopathogenic line of mumps virus, but showed little resistance to the cytopathogenic effect of vesicular stomatitis, Sendai, or Newcastle disease viruses.     

IMMUNITY IN MUMPS : II. THE DEVELOPMENT OF COMPLEMENT-FIXING ANTIBODY AND DERMAL HYPERSENSITIVITY IN HUMAN BEINGS FOLLOWING MUMPS

1. A specific antibody, demonstrable by the technique of complement fixation, regularly appears, or increases in concentration, in the sera of human beings during an attack of mumps or during convalescence. 2. Specific dermal hypersensitivity, demonstrable by the injection of heat-inactivated mumps virus, has been shown to develop in 6 human beings after recovery from mumps. 3. Complement-fixing antibody and the hypersensitive state also emerge as a result of clinically inapparent infection with the virus of mumps. 4. These two phenomena are apparently unrelated in respect to immunologic mechanisms. 5. The data presented indicate that the complement fixation test should prove of value both in diagnosis and in the determination of immunity. 6. The skin test for dermal hypersensitivity, on the other hand, becomes positive after recovery and therefore would appear to be useful only as an index of resistance.     

ACUTE ASCENDING MYELITIS FOLLOWING A MONKEY BITE,WITH THE ISOLATION OF A VIRUS CAPABLE OF REPRODUCING THE DISEASE

A case of acute ascending myelitis which followed the bite of an apparently normal Macacus rhesus monkey is described. The clinical course as well as the pathological changes has been studied and found to be suggestive of a virus cause for the disease. The absence of perivascular demyelinization removes the case from the realm of acute disseminated encephalomyelitis and establishes it more or less definitely as a primary acute infectious myelitis. An extremely important feature of the pathological picture of this disease has been the presence of focal necrosis in the viscera (spleen, adrenals, regional lymph nodes). Attempts to transmit the disease to Macacus rhesus monkeys, dogs, mice, and guinea pigs, employing glycerinated organs from the human case, proved unsuccessful. By the inoculations of rabbits the presence of a strongly neurotropic, filtrable virus was demonstrated in the patient''s brain, cord, and spleen. Following intracutaneous injection of it as derived either from brain and cord or spleen, an experimental disease develops in rabbits which strikingly resembles the human disease in the character of the local lesion, the incubation period, development of urinary retention, and flaccid paralysis of the posterior extremities with cephalad progression, death by respiratory failure, and finally by the occurrence of focal necrosis in the spleen, adrenals, and liver. In attempting to establish the identity of this virus, (the B virus), a consideration of its biological properties excludes the viruses of poliomyelitis, rabies, vaccinia, Virus III disease of rabbits, and the other viruses which are known to produce similar intranuclear inclusion bodies, except perhaps herpes. Although the relationship between the B virus and the virus of herpes must still be determined by cross-immunity tests it has been shown to possess certain properties which warrant consideration of it as a distinct entity.     

The estimated burden of 15 vaccine-preventable diseases from 2008 to 2020 in Japan: A transition by the COVID-19 pandemic

BackgroundEvaluating the national burdens across multiple vaccine-preventable diseases (VPDs) can be informative to identify the areas for improvements in the national immunization program.MethodsThe annual burden of diseases from 2008 to 2020 in Japan were calculated with the incidence- and pathogen-based approach for the 15 VPDs (hepatitis B virus infection, human papillomavirus (HPV), influenza, invasive pneumococcal disease, invasive Haemophilus influenzae type b (Hib) disease, invasive meningococcal disease, Japanese encephalitis, measles, mumps, pertussis, rotavirus, rubella, tetanus, tuberculosis and varicella), using disability-adjusted life year (DALY).ResultsThe average annual burden between 2008 and 2020 is the highest in influenza (114,129 DALY/year), followed by HPV infection, hepatitis B virus infection, tuberculosis and mumps (109,782, 69,883, 23,855 and 5693 DALY/year). In the pre-COVID-19 period (2008–2019), the decreasing trend of burden was observed in hepatitis B virus infection, invasive pneumococcal disease, invasive Hib disease, tuberculosis and varicella. HPV infection is the only VPD which had more than 100,000 DALY/year for all years during the study period. In 2020, the estimated annual burdens are decreased in influenza (71%), invasive pneumococcal disease (51%), invasive Hib diseases (54%), invasive meningococcal disease (64%), measles (98%), mumps (47%) pertussis (83%), rotavirus infection (95%), rubella (94%) and varicella (35%) compared with those in 2019.ConclusionsThe study demonstrated decreasing trends of burdens for some VPDs, while a persistently high burden has been observed for other VPDs, including HPV infection. The COVID-19 pandemic has caused dramatic reductions in the burdens of many VPDs in 2020.     

STUDIES ON THE INACTIVATION OF VACCINE VIRUS AND THE ACTION OF CERTAIN SUBSTANCES UPON THE INFECTING POWER OF THE INACTIVATED VIRUS

Vaccine virus, obtained from testicular inoculation shows a high susceptibility to chloroform as compared with ether, toluene, 95 per cent alcohol and acetone. Vaccine virus, after treatment with an amount of chloroform sufficient to render it incapable or only barely capable of originating an eruption in the rabbit''s skin, produces a characteristic eruption when injected with the supernatant fluid of embryonic tissue or sarcoma tissue "cultures" or kieselguhr, substances all of which are markedly irritative to the rabbit''s skin. Reactivation of the chloroformed vaccine virus is not possible when chloroform has been added to it in such quantity that the injection of large amounts of the treated virus fails to cause an eruption. Whenever reactivation has been accomplished it has been possible to get a vaccine eruption of greater or less intensity by the injection of large amounts of the chloroformed vaccine alone. Embryo and chicken sarcoma "culture" fluids when injected intradermally make the skin susceptible to the localization of the virus introduced intravenously. The bearing of these experiment on the interpretation of Gye'' theory of cancer causation is discussed.     

IMMUNITY IN MUMPS : IV. THE CORRELATION OF THE PRESENCE OF COMPLEMENT-FIXING ANTIBODY AND RESISTANCE TO MUMPS IN HUMAN BEINGS

Of 163 persons giving positive complement fixation tests who were exposed to mumps, 1 afterwards developed the disease; of 285 negative reactors similarly exposed, 56 afterwards came down with mumps. Of 78 individuals subjected to intimate exposure to mumps whose tests were originally negative and who failed to develop the disease, 41 per cent gave positive reactions when tested 1 month later. Seventy-seven per cent of complement fixation tests done on the sera of 565 normal adults who admitted a previous attack of mumps were positive. A similar correlation was recorded in tests on the sera of a small group of children with positive histories. Of 356 medical students admitting previous attacks, 80 per cent gave positive tests. Of 386 normal adults who denied previous attacks, 42 per cent gave positive tests; of 85 children giving negative histories, 38 per cent reacted positively. The results of complement fixation tests on the sera of 1665 normal adults (over 17 years) and 679 children (1 to 17 years) are recorded. It has been shown that 63 per cent of the adults and 57 per cent of the children had antibody in their blood which reacted with the virus of mumps. In groups in which exceptionally intense exposure was not known to have occurred in the past, the proportions of positive reactors were: adults, 61 per cent; children, 49 per cent. In contrast to these normal persons, the incidence of positive reactors among permanently institutionalized mental defectives was 38 per cent of 356 adults and 32 per cent of 475 children. In only 2 per cent of 320 normal adults and children did the titer of complement-fixing antibody reach 1–192. In no instance in which the endpoint was determined was a higher titer recorded. The results of complement fixation tests on the sera of mother and newborn infant were essentially the same in 5 instances.     

STUDIES OF THE HEMOLYSIS OF RED BLOOD CELLS BY MUMPS VIRUS : I. THE DEVELOPMENT OF MUMPS VIRUS HEMOLYSIN AND ITS INACTIVATION BY CERTAIN PHYSICAL AND CHEMICAL AGENTS

The conditions for the production of extra-embryonic fluids with hemolytic activity from chick embryos infected with mumps virus have been investigated. Infected fluids with strong hemolytic activity can be obtained by harvesting the fluids of 6- to 8-day-old chick embryos inoculated by the amniotic route after 5 to 6 days' incubation at 35°C. Under such circumstances, the hemolytic capacity of amniotic fluids is usually much higher than that of the allantoic fluids. The hemolytic activity and infectivity of the virus have been found to be reduced or destroyed by heat, formaldehyde, and ultraviolet irradiation under conditions which leave the hemagglutinating capacity practically unchanged. Ultraviolet irradiation appeared to have a greater deleterious effect on the infectivity of the virus than on its hemolytic capacity. The marked reduction or destruction of hemolytic activity of the virus produced by certain treatments with these various agencies was not accompanied by loss of the ability of the virus to elute following its adsorption on red blood cells during the process of hemagglutination. This test for hemolytic activity, which measures a more labile property of the virus than do determinations of virus hemagglutination or virus elution, may be useful in detecting changes which occur early during degradation of the virus.     

STUDIES ON THE MECHANISM OF IMMUNITY IN TYPHUS FEVER : II. ALLERGIC AND TOXIC REACTIONS PRODUCED WITH RICKETTSIA PROWAZEKI

The intradermal inoculation of Mexican typhus virus into immune guinea pigs produces a local reaction which is similar in its appearance to the lesion observed in the skin of normal animals submitted to the same treatment. The reaction in the immune animal appears earlier and fades sooner than the lesion in the normal guinea pig. The inoculation of heat-killed or formalin-killed Rickettsiae produces no significant reactions at the site of the intradermal injection in typhus immune guinea pigs. The virus, inoculated intradermally, has been recovered from the local lesion 72 hours after the injection into typhus immune guinea pigs. Normal guinea pigs and persons without a history of typhus fever present a congestion and some swelling of the skin at the site of the intradermal injection of formalinized Mexican Rickettsiae. The reaction appears 24 hours after the inoculation and fades within 48 hours. Heating the formalinized Rickettsia suspensions at 70°C. for 30 minutes renders them inactive in normal men and guinea pigs. From the experiments reported in this paper it seems that the reactions observed in typhus immune guinea pigs submitted to a second inoculation of typhus virus, belong to the group of reactions presented by tuberculous animals (Koch''s phenomenon) and the accelerated takes shown by immune persons submitted to revaccination with vaccinia virus. A heat labile substance has been demonstrated in the formalinized Rickettsia bodies, which produces a reaction in the skin of normal men and guinea pigs.     

THE MULTIPLICATION OF THE VIRUS OF MEXICAN TYPHUS FEVER IN FLEAS

The virus of Mexican typhus has been shown to multiply abundantly in the following species of fleas: Xenopsylla cheopis, Ceratophyllus fasciatus, Leptopsylla musculi, Ctenocephalus canis, Ctenocephalus felis. In all fleas, Rickettsia prowazeki was demonstrated within the epithelial cells of the stomach and within the cells of the Malpighian tubules. Whereas in infected lice enormous numbers of these organisms are discharged from the disintegrating cells into the intestinal content, only few Rickettsiae are found in the lumen of the fleas'' intestines. They are held back by the peritrophic membrane, which covers the mucosa of the entire stomach. Rickettsiae seem to enter the lumen of the gut almost exclusively by the route of the Malpighian tubules. Observations were made which seem to indicate that the fleas recover from the infection and that they are able to regenerate the partly destroyed intestinal mucosa. An explanation is given for the relative harmlessness of fleas as vectors of typhus.     

THE NATURAL HISTORY OF HUMAN POLIOMYELITIS : II. ELIMINATION OF THE VIRUS

Studies on the elimination of virus in human paralytic poliomyelitis during the first 2 weeks of the disease, revealed the following:— 1. The nasal (not nasopharyngeal) secretions collected from 22 patients on cotton plugs over a period of 3 days and the saliva and oral secretions expectorated during a similar period by 20 patients failed to yield virus. 2. In 10 of the patients whose secretions (nasal, oral, or both) were investigated, virus was isolated from single specimens of the lower intestinal contents. 3. No virus was found in large amounts of urine (up to 200 cc.) obtained from 12 patients, 6 of whom had paralysis of the bladder. 4. In the present tests virus was found 4 times more often in the stools of patients under 8 years of age (64 per cent of 11 cases) than in older individuals (17 per cent of 12 cases). This difference was found to obtain when our data were combined with those which could justifiably be selected from the literature, the total figures indicating that virus has been isolated from 50 per cent of 58 children under 8 years of age and from 12 per cent of 60 older individuals. 5. No support was found for the hypothesis that poliomyelitis virus in the stools originates from swallowed nasal secretions and saliva or oral secretions.