Temperature-sensitive mutants of influenza A virus. XII. Safety, antigenicity, transmissibility, and efficacy of influenza A/Udorn/72-ts-1[E] recombinant viruses in human adults.

The influenza A/Hong Kong/68-ts-1[E] virus (shutoff temperature, 38 C), which possesses many characteristics desirable in a vaccine virus, was used as a donor of its two temperature-sensitive (ts) lesions to the antigenically divergent influenza A/Udorn/72 wild-type virus. Two subsets of Udorn/72-ts-1[E] recombinant viruses were evaluated in seronegative volunteers (serum titer of hemagglutination-inhibiting antibody, less than or equal 1:8). The first subset, represented by clone 13, possessed a shutoff temperature of 39 C and only one of the two ts lesions; this virus was insufficiently attenuated for use in humans.

Temperature-sensitive mutants of influenza A virus. XI. Transfer of ts lesions in the Hong Kong/68-ts-1 [a] virus to the influenza A/Udorn/72 wild type.

The presence of the temperature-sensitive (ts) lesions of complementation-recombination groups 1 and 5 in the Hong Kong/68-ts-1[A] virus was confirmed by genetic analysis of ts recombinants of the Hong Kong/68-ts-1[A] virus and a Udorn/72 wild-type virus. Three classes of Udorn/72-ts recombinants were found. One class possessed both ts genes of the Hong Kong/68-ts-1[A] parent, a second class possessed the ts lesion characteristic of group 1, and a third class possessed the ts lesion of group 5.

Antigenic relationships among five reovirus-like (RVL) agents by complement fixation (CF) and development of new substitute CF antigens for the human RVL agent of infantile gastroenteritis.

The human reovirus-like (HRVL) agent, Nebraska calf diarrhea virus (NCDV), epizootic diarrhea of infant mice (EDIM) virus, simian agent (SA)-11, and the "O" (offal) agent were found to be similar, if not identical, in reciprocal complement fixation (CF) tests employing hyperimmune animal sera. In addition, in CF tests with paired sera from 35 diarrhea patients who shed the HRVL agent, 74% developed serologic evidence of infection with the HRVL antigen, 43% with NCDV, 51% with EDIM virus, 57% with SA-11, and 71% with the "O" agent. Thus, in addition to the NCDV, which had previously been described as a suitable substitute CF antigen for the HRVL agent, the SA-11, "O", and EDIM viruses may also be utilized as substitute antigens for the HRVL agent.

Use of temperature-sensitive mutants of influenza A virus as live virus vaccine strains. Evaluation in laboratory animals, adults and children.

Temperature-sensitive (ts) recombinants of influenza A virus were evaluated for use in a live virus vaccine. Evidence from several sources suggested that the ts lesions were responsible for attenuation of these mutants. Specification of attenuation by defined genetic lesions which can be assayed for in the laboratory offers an advantage to the use of ts viruses for vaccination.

Human reovirus-like agent as the major pathogen associated with "winter" gastroenteritis in hospitalized infants and young children.

We found a human reovirus-like agent in the stools of 42 per cent of 143 infants and young children hospitalized with acute gastroenteritis between January, 1974, and June, 1975. Half the patients studied by electron microscopy and serologic technics had evidence of infection with the agent. The infection had a seasonal pattern: 59 per cent of those admitted during the cooler months (November to April) shed the agent, with a peak of 78 per cent in December, 1974, and January, 1975, combined.

Temperature-sensitive mutants of influenza A virus: response of children to the influenza A/Hong Kong/68-ts-1(E) (H3N2) and influenza A/Udorn/72-ts-1(E) (H3N2) candidate vaccine viruses and significance of immunity to neuraminidase antigen.

One of two slightly different influenza A/ts-1[E] recombinant candidate live vaccines was given intranasally to each of 23 young children. Twelve of 15 children who had no serum HI antibody but who did have serum ANAB at the time of administration became infected and 1 had mild rhinitis. All eight who lacked both types of antibody became infected and they shed virus in higher titer and for longer than the former group; five had rhinorrhea and five had mild fever.

Experimental production of respiratory tract infection with Mycoplasma pneumoniae in rhesus monkeys.

Knowledge of the pathogenesis of pneumonia due to Mycoplasma pneumoniae has been derived primarily from experimental infection of rodents. As part of an effort to establish a model with a closer resemblance to man, three seronegative, young, adult rhesus monkeys (Macaca mulatta) were inoculated with M. pneumoniae (10(7.

Growth and genetic stability of the ts-1 mutant of respiratory syncytial virus at restrictive temperatures.

An in vitro study was performed to define in greater detail those factors which favored the growth of the ts-1 mutant of respiratory syncytial virus under restrictive conditions and the emergence of genetically altered virus with decreased temperature sensitivity. Replication of ts-1 occurred at each of the restrictive temperatures of 37, 38, and 39 C, even through plaque formation was not observed. The level of virus growth under restrictive conditions was inversely related to the incubation temperature and directly related to the multiplicity of infection.

Cell-mediated immunity to respiratory syncytial virus induced by inactivated vaccine or by infection.

Transformation and increased mitotic activity in donor lymphocytes exposed to specific antigens is considered by many to be a manifestation of cell-mediated immunity. In attempts to understand the apparent "sensitization" of individuals to respiratory syncytial virus (RSV) as a result of receiving inactivated RSV vaccine, in vitro lymphocyte transformation studies were carried out on infants who had received inactivated RSV vaccine and on infants who had received a similarly prepared inactivated African green monkey kidney (AGMK) cell-grown parainfluenza type 1 virus vaccine or a trivalent parainfluenza vaccine prepared in hen's eggs. Each group included some infants who had, and others who had not, undergone natural RSV infection under our observation before the lymphocyte studies.

Nonbacterial gastroenteritis.

Significant progress has been made toward determining the agents of acute, infectious, nonbacterial gastroenteritis. Two distinct types of viruses have been implicated. One of these, a particle about 27 nm in diameter, is involved in the acute epidemic form of the disease of short duration.

Induction of diarrhea in colostrum-deprived newborn rhesus monkeys with the human reovirus-like agent of infantile gastroenteritis.

Diarrhea developed in five newborn rhesus monkeys (Macaca mulatta) inoculated orally on the first day of life with the human reovirus-like agent of infantile gastroenteritis. Incubation period ranged from 2-5 days; virus particles were detected in stools in association with illness, and virus shedding lasted between 1 and 3 days. Virus derived from monkeys that developed illness following inoculation was infectious for other monkeys but did not induce diarrhea which could be associated temporally with virus shedding.

Recent advances in the aetiology of viral gastroenteritis.

Studies with the human reovirus-like (HRVL) agnet, also designated rotavirus and duovirus, have revealed that it is a major aetiological agent of diarrhoea of infants and young children in many parts of the world. In a study of patients admitted with a diarrhoeal illness to the Children's Hospital of the District of Columbia in the United States from January 1974 to June 1975, it was found that half of the patients studied by both virus shedding (by electron microscopy) and serological (complement-fixation) techniques demonstrated evidence of infection with the HRVL agent. The temporal distribution of infections with the HRVL agent followed a seasonal pattern with this agent being shed exclusively by patients admitted during the cooler months of the year.

Pattern of shedding of the Norwalk particle in stools during experimentally induced gastroenteritis in volunteers as determined by immune electron microscopy.

In 11 of 23 volunteers the Norwalk virus-like particle was visualized by immune electron microscopy in at least one stool specimen obtained during the acute phase of experimentally induced nonbacterial gastroenteritis. Examination of multiple stool specimens obtained during the course of illness in these 11 volunteers revealed maximal concentration of Norwalk virus-like particle at the onset of illness and shortly thereafter; in no case was the Norwalk particle visualized in stools obtained before the onset of illness. This finding further suggests that the Norwalk particle was the etiological agent of the Norwalk gastroenteritis outbreak.

Failure of attenuated temperature-sensitive influenza A (H3N2) virus to induce heterologous interference in humans to parainfluenza type 1 virus.

The present investigation was undertaken to determine if a candidate live vaccine virus, influenza A/Hong Kong/68-ts-1 [E] (H3N2), induced heterologous interference against an interferon-sensitive, wild-type, parainfluenza type 1 challenge virus. The parainfluenza virus was administered 7 days after Hong Kong/68-ts-1 [E] virus infection. The clinical response, daily quantitative virus shedding, interferon production, and serum and nasal wash antibody responses were determined in an experimental group (influenza A virus followed by parainfluenza virus) and 10 volunteers in a control group (parainfluenza virus only).

Transmission of Creutzfeldt-Jakob disease to the stumptail macaque (Macaca arctoides).

The successful transmission of Creutzfeldt-Jakob disease from both affected human and chimpanzee brain to stumptail macaques has been accomplished. The incubation period of 5 yr was the same for both animals; however, the course of the disease was longer in the animal receiving the human brain. In both cases, initial mild symptoms slowly remitted only to reappear some 4 mo later.

New complement-fixation test for the human reovirus-like agent of infantile gastroenteritis. Nebraska calf diarrhea virus used as antigen.

A complement-fixation (C.F.) test for the human reovirus-like agent of infantile gastroenteritis has been developed using the serologically related Nebraska calf diarrhoea virus (N.

Potential of attenuated respiratory syncytial virus vaccine for infants and children.

Respiratory syncytial virus (RSV) disease is a major cause of death and hospitalization in infancy and a frequent cause of morbidity throughout childhood. Serum antibody does not protect as is evident from the study of natural disease and use of killed vaccines. Local antibody responses occur in natural illness.