Experimental diabetes in mice infected with Coxsackie viruses

The influence of Coxsackie B4 and AI3 viruses on the pancreas of mice (resistant and susceptible to diabetes) was studied. Glucose intolerance and changes in the synthesis of immunoreactive insulin were detected in all the treated groups of animals. Biochemical changes were more prominent in male DBA/2 mice, infected with Coxsackie B4 virus, in FI (CBA X C57Bl/6) hybrids and in female DBA/2 mice infected with Coxsackie AI3 virus and alloxan.     

Salivary Excretion of Coxsackie B-1 Virus in Rabbits

Coxsackie B-1 virus was injected into the ear vein of albino doe rabbits. Saliva and blood samples were taken before the injection of virus and at specific times thereafter. Virus was recovered in the whole saliva when the blood titer was approximately 10(4) TCID(50) per 0.1 ml or greater. The virus could be detected in the saliva as early as 2 min after the initiation of the viremia. The recovered virus was shown to be the same as the injected virus by serological identification of the recovered virus with neutralizing antibody for Coxsackie B-1 virus. These results suggest that virus may be transmitted to other animals in the saliva of animals who are in the viremic phase of infection without infection of the oropharyngeal tissues.     

Enteroviral Syndromes in Toronto, 1964

Virological or serological investigations of 72 children in Toronto and environs, who were hospitalized between January and October 1964 with a variety of syndromes, revealed evidence of enteroviral infection in 29 subjects. Coxsackie B2 was the dominant enterovirus, being isolated from feces and/or cerebrospinal fluid (CSF) of three children with aseptic meningitis, three with pleurodynia, one with myalgia and one with pericarditis; four additional patients showed rising antibody titres to this virus. Coxsackie B1 virus, which has not been isolated in Toronto since 1950, was recovered from feces of three patients with pleurodynia, CSF of one patient with myalgia, and peritoneal fluid of a child with primary peritonitis; one patient with pericarditis showed a rising antibody titre to Coxsackie B1 virus. Coxsackie B3, B4 and Echo 23 viruses were associated with one case each of pleurodynia. Coxsackie B5 virus infected five patients with aseptic meningitis, and one each with pericarditis and myocarditis.     

Susceptibility of cell lines of primate and nonprimate origin to certain human viruses

A comparative study was made of the susceptibility of 11 cell lines of human and animal origin, the WI-38 cell strain and fresh cultures of human thyroid, monkey kidney and hamster embryo tissues to certain human viruses. The animal cell lines were derived from monkey, rabbit, mouse, pig and calf tissues. The viruses used were strains of influenza A₂ and B viruses, parainfluenza viruses types 1, 2 and 3, RS virus, adenoviruses types 3, 4 and 21, poliovirus type 1 and Coxsackie A type 21 and Coxsackie B type 3 viruses. Cell lines derived from nonprimate tissues were generally less susceptible than cell cultures of human and simian origin. The combined use of fresh cultures of human thyroid and monkey kidney tissues and of a human cell line seems to provide a satisfactory indicator system for the viruses employed in this study.     

Host selenium status selectively influences susceptibility to experimental viral myocarditis

The purpose of the present work was to determine whether dietary selenium (Se) deficiency could influence the injurious effect of human viruses other than Coxsackie virus B3 (CVB3) on mouse heart. Weanling C3H/HeN mice were fed a Se-deficient or Se-adequate diet for 4 wk and then were inoculated intraperitoneally with one of the following viruses: Coxsackie virus B1 (CVB1), echovirus 9 (EV9), Coxsackie virus A9 (CVA9), or herpes simplex 1 (HSV1). Polio virus 1 (PV1) was employed as a negative control. Prior to inoculation, mean serum Se levels were 430 versus 61 ng/mL in adequate versus deficient mice, respectively. Ten days later, hearts were removed and processed by routine histological procedures. Cardiac lesions were scored according to the number and size of myocarditic foci. Significantly greater heart damage resulting from CVB1 and EV9 was observed in Se-deficient than in Se-adequate mice, and the Se status had no influence on CVA9-induced myocarditis. In contrast, heart damage caused by HSV1 was significantly milder in Se-deficient than in Se-adequate mice. Therefore, it may be concluded that the Se status of the murine host selectively influences the degree of viral-induced myocarditic lesions.     

Replication of diabetogenic and nondiabetogenic variants of encephalomyocarditis (EMC) virus in ICR Swiss mice

The replication of the diabetogenic D (EMC-D) and nondiabetogenic B (EMC-B) variants of encephalomyocarditis virus in various tissues of the murine host was determined. Pancreatic insulin levels were also measured. EMC-D replicated in the spleen, pancreas, heart, lung, and intestines, while EMC-B was limited to the spleen and pancreas. EMC-B interfered with the replication of EMC-D in each of the tissues examined. Insulin levels were initially increased by both viruses. By 4 days postinfection, insulin levels were either normal or undetectable in (EMC-D)-infected animals, but were dramatically elevated in those infected with EMC-B.     

Modelling coxsackie-virus infection in pregnant mice in long-term experiment

The effect of virus infection on the organism of pregnant mice and their posterity was studied in experiment. The animals were infected with the prototype strain A13 (Flores) of Coxsackie virus which was administered on days 4, 7, 11, 15 and 19 of pregnancy. It has been demonstrated that pregnant mice are much more sensitive to the virus than non-pregnant females and that the placenta, along with the striped muscles, is the main reservoir of the virus. The obtained results also suggest that the virus penetrates into the tissues of the embryo most intensively in the second half of pregnancy and that the duration of manifestation of the virus in the tissues of the embryo depends on the period of intrauterine development. The study of the model system (long-term organ culture of the tissues of the organism) enabled us to establish a new fact of the cytoproliferative activity of A13 Coxsackie virus in the organ culture and in the placenta of mice infected in vivo, and also in the organ culture of the liver of their newborn, which is important for the confirmation of clinical observations presuming a high probability of intrauterine infection by the mentioned virus with its subsequent protracted persistence in the organism of the newborn.     

Immunity status against coxsackie viruses B in 25 patients with acute myocarditis

Viruses are an important cause of myocarditis, particularly the enterovirus group B coxsackievirus. Viral infection may be suspected on the basis of history and presentation and can be proved by direct or serological identification of virus. Twenty-five patients were diagnosed with acute myocarditis and were investigated with a serologic test battery covering Coxsackie viruses group B types 1 to 5 at the National Reference Center for Enteroviruses in Cantacuzino Institute Bucharest, Romania. A possible Coxsakie B virus etiology could be documented in 11 from 25 cases with acute myocarditis and high titers against Coxsackie virus B type 2 (1 patient), type 3 (5 patients) and type 5 (in 4 patients) were detected. In one HIV positive patient (17 years old), a concomitant infection with Coxsackie virus B types 2 and 4 was detected. The earlier detection of enterovirus myocarditis could be followed by antiviral therapies with a potential therapeutic role.     

Experimental therapy of Coxsackie B3 virus infection with immunomodulator contained in water-soluble Penthaphylloides fruticosa extract

The possibility of using water-soluble substance extracted from P. fruticosa in experimental Coxsackie B3 virus infection in newborn mice was studied. The curative action of the extract was studied by its injection to the infected animals for 7 days and its prophylactic action--by introducing it 24 hours and, repeatedly, 1 hour before infection. The study revealed the increase of such characteristics as survival rate, mean survival time, body weight dynamics, as well as decreased level of virus accumulation in the brain, liver and heart tissues of the animalsunder experiment. The immunomodulating properties of the extract were shown.     

Viral Infections of Toronto Children During 1965: I. Enteroviral Disease

Enteroviruses were isolated from feces and/or cerebrospinal fluid of 29 of 43 Toronto children who contracted aseptic meningitis, pleurodynia, abdominal pain or febrile upsets between June and October, 1965. Coxsackie A9 virus was the dominant agent in aseptic meningitis and Coxsackie B1 virus in pleurodynia and other syndromes. Sero-logical evidence of recent Coxsackie B1 and Echo 6 infection was obtained in two additional patients with aseptic meningitis who did not yield virus, and elevated Coxsackie B1 antibody titres were found in one patient with pericarditis. A newborn infant died with myocarditis due to Coxsackie B1 virus following infection of the mother during the immediate antenatal period. Paired sera collected only two to four days apart from patients with enteroviral syndromes or mumps meningoencephalitis frequently showed four-fold or greater increases of antibody levels.     

Rash Associated with Coxsackie A9 Infection

Coxsackie A9 virus was identified by the authors during the fall of 1965 in Montreal in six children with fever and exanthem. Three of the six children were siblings. The exanthem was centrally distributed as described by Lerner et al. and consisted of discrete maculopapules 3 to 4 mm. in diameter. The viral agent was recovered and identified in tissue culture in five cases, while in the sixth Coxsackie type-A lesions were produced in suckling mice. Serological confirmation was obtained in two patients from whom sera were available. In contrast, no exanthem was observed in three older patients with a diagnosis of aseptic meningitis associated with Coxsackie A9 virus. In only one of 16 patients with Coxsackie B virus infection was an exanthem observed during the same period.The true incidence of Coxsackie A9-associated exanthems is difficult to determine because of the benign nature of the disease.     

Participation of Coxsackie B viruses in the occurrence and recurrence of infectious-allergic myocarditis (a serological study)

The serological study of sera from patients with infectious allergic myocarditis and from healthy persons has revealed essential differences in the occurrence and titers of antibodies to Coxsackie virus B. In patients with infectious allergic myocarditis the infectious process has been found to be significantly more frequently linked with Coxsackie viruses B, serovars 2 and 4.     

Biochemical changes induced by Coxsackie B4 virus in short-term culture of mouse pancreatic islets

Isolated mouse pancreatic islets were infected in vitro with two strains of Coxsackie B4 virus — a tissue culture - adapted strain and a mouse pancreas-adapted strain. Within 48 h of infection changes had occurred in the biochemical activities of islets infected with the mouse pancreas-adapted strain of virus. Basal insulin release was increased two-fold in these islets, while glucose-induced insulin secretion remained unchanged. Insulin biosynthesis was greatly reduced at a sti, mulatory concentration of glucose (20 raM), thus leading to a reduced insulin content in these islets. These effects are of importance because they demonstrate that certain strains of Coxsackie B4 virus, like encephalo-myocarditis virus, may selectively alter B-cell function in vitro.     

Viral Antibodies in Diabetes Mellitus

Sera from 123 patients with diabetes mellitus of recent onset, 155 patients with diabetes of more than two years'' duration, and 250 normal persons were collected over a period of two and a half years. All sera were tested for neutralizing antibody to Coxsackie virus types B1–6, and a sample was tested for complement-fixing antibody to a number of viral, rickettsial, and mycoplasmal antigens.In diabetics of recent onset no evidence was found of any excess of antibodies to mumps virus or some common respiratory viruses. Insulin-dependent diabetes within three months of onset were found to have higher antibody titres to Coxsackie B virus, particularly of type B4, than either normal subjects or patients with diabetes of longer duration.     

Diabetes in Mice after Coxsackie B4 Virus Infection

The intraperitoneal inoculation of CD₁ mice with Coxsackie virus B₄ resulted in the raising of blood sugar levels to diabetic values 12 days after the administration of the virus. Serum insulin remained inappropriately low. Light microscopy changes in the islets of Langerhans showed mononuclear cell infiltration of the islets and marked degranulation of the β cells. The acinar tissue appeared to be little changed. It is concluded that Coxsackie B₄ virus may cause a diabetic state compatible with islet cell damage.     

Coxsackievirus in an infant chimpanzee.

Coxsackie B viruses may cause a severe, often fatal, illness in newborn and infant human subjects. As recorded in this case, infant chimpanzees respond similarly to Coxsackie B-5 virus.     

Effect of poly(I).poly(C12U) (Ampligen) on enteric virus (rotavirus, poliovirus and Coxsackie B3 virus) infections

The effects of poly(I) poly(C₁₂U) (Ampligen) on infections with enteric viruses (rotavirus, poliovirus and Coxsackie B3 virus) were studied in vitro. Ampligen exhibited antiviral activity against rotavirus, especially when treatment was performed prior to inoculation of the virus. It was partially effective against Coxsackie B3 virus, but not against poliovirus. It is suggested that the observed effects may be due to the production of interferon induced by Ampligen.     

Isolation of Enteric Viruses in Ontario During 1960-1962

During the past three years at the Central Laboratory, Ontario Department of Health, 681 isolations were made in tissue culture from 6822 specimens submitted for virus studies by physicians and hospitals from all over Ontario. Nearly 74% of the isolates were enteroviruses, approximately 5% adenoviruses and about 1% reoviruses. The remaining 20% are still to be identified.Although the bulk of isolations was made during the same three-month period (August, September and October) of each year, the predominant virus types varied from year to year. Poliovirus 1 was most commonly encountered in 1960, Coxsackie B5 in 1961 and Echo 9 in 1962.Among other types isolated in smaller numbers were Coxsackie A1, 9 and 10, Coxsackie B1, 2, 3 and 4, Echo 1, 2, 5, 6, 7, 8, 11, 14, 17, 18 and 19, Reovirus 1, 2 and 3, Adenovirus 1, 2, 3, 4, 5, 7 and 16, as well as Frater-type virus. Most of these types were isolated for the first time in Ontario and represent additions to the existing list of viruses known to occur in this province.     

Immunomodulation of encephalomyocarditis virus-induced disease in A/J mice.

The E variant of encephalomyocarditis (EMC) virus causes an encephalomyelitis and coagulative necrosis of the pancreas and parotid glands in some but not all strains of inbred and outbred mice. In other models of disease caused by picornaviruses, depletion of specific lymphocyte subsets abrogates the development of tissue lesions. In this study, severe encephalomyelitis and acinar pancreatitis and parotitis developed in adult male A/J mice infected with 100 PFU of EMC virus. Depletion of the CD4+ subset of T lymphocytes in vivo with a monoclonal antibody (MAb) prior to EMC virus inoculation protects mice from developing encephalomyelitis, pancreatitis, and parotitis. This effect is also seen when animals are treated with anti-CD4 and anti-CD8 in combination, but the anti-CD8 MAb alone does not ameliorate the disease. Overall, concentrations of virus in tissues from anti-CD4-treated animals are lower than in immunologically intact control mice. Small-plaque variants of virus were also recovered from the tissues in some animals in this group. CD4+ lymphocytes are involved in the expression of EMC virus-induced pancreatitis and parotitis in A/J mice. This specific subset of T cells would appear to influence EMC viral tropism or replication in various organs.     

Selenite inhibition of Coxsackie virus B5 replication: implications on the etiology of Keshan disease.

Keshan disease is a cardiomyopathy of unknown origin reported in some areas of China. Because of epidemiologic features, this disease was ascribed to an infectious agent, likely a Coxsackie virus, but it has also been thought to depend on selenium deficiency, mainly because selenite is effective in its prophylaxis. We examined the hypothesis that pharmacological activity of selenite on Coxsackie virus growth was associated with prevention of Keshan disease. We studied the antiviral effects of three selenium compounds on Coxsackie virus B5 replication: five microM selenite reduced viral replication, whilst 10 microM selenate and selenomethionine did not exhibit any antiviral activity. The inhibitory activity of selenite on viral replication was due to its toxicity following its interaction with thiols, as that activity could be blocked by dithiothreitol, a sulfhydryl-protecting agent known to reverse several toxic effect of selenite. Zinc, another inhibitor of selenite toxicity, also counteracted the antiviral effect of selenite. The selenium compounds showed only limited activity against herpes simplex 1 virus and IHD strain of vaccinia virus. A direct inhibitory effect of selenite on Coxsackie virus replication might explain the efficacy demonstrated by this compound in the prophylaxis of Keshan disease.