Split Virus Research Articles

Electroelution for purification of influenza A matrix protein for use in immunoassay

A new preparative method for isolation of matrix protein from type A influenza virus was developed. Commercially available whole virus or split virus vaccines were lysed, and the soluble proteins separated by electrophoresis on polyacrylamide gel. The matrix protein was located on the gel by precipitation with KCl, and recovered by electroelution. The method was technically simple and required little direct supervision during the two-step recovery process. Yields of A matrix were consistently high, averaging 68.1% in five trials with A/Brazil/X-71. The method was also successful with other A viruses, although not with influenza B virus. Isolated A matrix had less than 0.5% contamination by hemagglutinin or nucleoprotein, as determined by immunoblotting and ELISA, Matrix protein was immunoreactive in Western blots and was detectable in concentrations as low as l ng/ml with ELISA. The isolated matrix provided a suitable standard for detection of matrix protein in nasal washes from patients with influenza A virus infection, and could also be used to detect anti-matrix antibodies, including monoclonal antibodies in tissue culture supernatants. The advantages of electroelution for separation of matrix protein compared to other methods were its technical simplicity, applicability to formalin-fixed influenza virus in commercially available vaccines, its consistently high yield, and its very high level of purification. influenza matrix electroelution polyacrylamide gel electrophoresis

The effect of split virus influenza vaccination on theophylline pharmacokinetics.

The effects of split virus influenza vaccine on the pharmacokinetics of theophylline and the in vivo and in vitro induction of interferon were studied in 23 volunteers. No effect on the total body clearance, half-life, or mean residence time of theophylline was found as a result of influenza virus vaccine. Additionally, no interferon activity was detected in vivo for as long as 24 h after vaccination. Finally, no production of interferon occurred in tonsil or peripheral lymphocyte cultures inoculated with split virus vaccine. We conclude that split virus influenza vaccine does not affect theophylline pharmacokinetics. In addition, we find no evidence that split virus influenza vaccine functions as an interferon inducer in humans.

Split virus influenza vaccination in children: an evaluation of efficacy.

Twenty-seven children aged between 4 and 13 years were given two injections of a split virus influenza vaccine (MFV-Ject, Institut Merieux) at a dose of 0.5 ml and an interval of 1 month. Each vaccination contained A/Philippines/2/82 (H3 N2) 10 micrograms HA, A/Brazil/11/78 (H1 N1) 10 micrograms HA, B/Singapore/222/79 15 micrograms HA. The overall serological response was good although the B group virus produced slightly less response; however, no children were seropositive to B prior to vaccination, reflecting a low previous exposure. Children aged over 10 years showed a generally higher serological response. Side-effects were minimal. A favourable reactogenicity/efficacy ratio was found.

Persistence of Antibody After the Administration of Influenza Vaccine to Children with Cancer

Children with cancer who received an intramuscular injection of inactivated split or whole bivalent (A/New Jersey/76-A/Victoria/75) influenza virus vaccine during the fall of 1976 were tested up to 18 months later for the persistence of antibody. Titers of antibody greater than of equal to 10 to influenza A/New Jersey/76 virus were present in the sera of 34 children two weeks after immunization. The geometric mean titer was 40.0. Eighteen months later a titer greater than or equal to 10 was present in 13 (38%) children; the geometric mean titer was 8.5. Of 36 children with a titer of antibody greater than or equal to 10 to A/Victoria/75 virus two weeks after immunization, 34 still had a detectable titer in the later examination. The geometric mean titer declined from 65.3 to 25.6. Titers of antibody to B/Hong Kong/72 virus (not included in the vaccine) remained relatively stable. In 11 children the usual chemotherapy was discontinued during the study. The shorter persistence of antibody to A/New Jersey/76 virus, in contrast to that of the other two influenza viruses tested, was associated with a lack of prior exposure to the virus and the absence of subsequent natural infection with this virus or an antigenically related subtype. The potential effect of cancer chemotherapy on the persistence of antibody levels is unclear.

Inadvertent Intravenous Administration of Trivalent Influenza Vaccine

INFLUENZA vaccine is administered annually to millions of persons in order to prevent influenza virus infection.¹The Public Health Service recommends administration of trivalent influenza vaccine (AH1N1, AH3N2, and B) to high-risk patients in the late summer and fall months.²The package inserts that accompany influenza vaccine and the manual<i>Facts and Comparisons</i>(St Louis, Facts and Comparisons Inc, 1980) advise that the vaccine should not be administered intravenously (IV). However, in the discussion of adverse reactions, the possible consequences of IV injection of the influenza vaccine are not mentioned. We report here the misidentification of a trivalent influenza vaccine cartridge for a heparin flush cartridge and subsequent IV administration of trivalent influenza vaccine to at least one patient. <h3>Report of a Case</h3> On Oct 22, 1979, influenza vaccine was ordered for three patients in one hospital ward. Three cartridge-needle units (TUBEX) (0.5 mL) of trivalent (split virus)

Zur lokalen und systemischen Immunantwort nach Influenza- Schutzimpfung mit drei verschiedenen Vakzinetypen

Three types of inactivated bivalent influenza vaccines containing the A/Victoria/3/75 (A-Component) and B/Hongkong/8/73 (B-Component) antigens either as aluminium salt adsorbed whole virus (AlorbatTM, "A") and split virus (Begrivac STM, "B") or as fluid subunit (SandovacTM, "S") products were given to immunological homogeneous study groups of adults and children. The immune response was compared between the groups on the basis of systemic and local neuraminidase inhibiting (NI) antibody induction 4 weeks after vaccination. In evaluation of the serologic results the sea and nasal washings were tested in the ESSEN-NIT with use of the A/Eq(eq1)-VIC (N 2) recombinant antigen and the B/Hong kong/8/73 strain. The number of individuals (adults plus children) evaluated ranged from 33 to 35 per vaccine. The percentages of persons greater than or equal to 16 years old showing a significant (1.5 x, or greater) response to the A-component were 100%, 96% and 90%, respectively for the vaccine "A", "S" and "B"; the values for the B-component were 70%, 79% and 74%, respectively. The observed conversion rates in children ranged 70% to 100%. Local antibody formation was less pronounced, the percentages of significant conversions were scattered over a wide range from 13% to 80% yielding slightly better responses in children than adults. Taken altogether, the results of the trial indicate that the 3 vaccines studied induced appreciable concentrations of systemic antibodies to the neuraminidase antigen without significant differences between the vaccine types. This potency of the influenza vaccines to induce high titers of NI antibodies thus provides another marker for their immunogenicity and conceivably for their efficacy.

The specificity of the anti-haemagglutinin antibody response induced in man by inactivated influenza vaccines and by natural infection.

The anti-haemagglutinin antibody response in adult human volunteers to inactivated whole virus or tween ether split influenza A/Victoria/75 (H3N2) and A/Scotland/74 (H3N2) virus vaccines was investigated using antibody absorption and single-radial-haemolysis (SRH) techniques. The concentrations of haemagglutinin (HA), nucleoprotein (NP) and matrix (M) antigens measured by single radial diffusion (SRD) and rocket immunoelectrophoresis were similar for both the whole virus and split vaccines. Whole virus and split vaccines induced crossreactive (CR) antibody in 87% of vaccinees. Strain specific (SS) antibody to A/Hong Kong/1/68 of the homologous virus was induced less frequently than CR antibody. Higher anti-haemagglutinin antibody titres were detected in persons receiving the split virus vaccines than in those receiving the whole virus vaccines. No antibody to the type-specific matrix protein was detectable, but 33% of volunteers developed an antibody rise to type-specific nucleoprotein antigen. The specificity of the anti-haemagglutinin antibody response in human adults to natural infection with A/Port Chalmers/73 (H3N2) virus was similar to that induced by inactivated vaccines in that a high proportion of subjects developed CR anti-haemagglutinin antibody, which reacted with A/Hong Kong/68 virus and the homologous A/Port Chalmers/73 virus, and SS antibody for A/Hong Kong/68 virus but SS antibody for A/Port Chalmers/73 virus was infrequently stimulated by natural infection.

The relative immunogenicity in mice of whole and split influenza virus.

The relative immunogenicity in mice of whole influenza virus, and virus split with different disrupting agents, was compared. Using the single radial immunodiffusion test to estimate the haemagglutinin antigen concentration in different virus preparations, it was found that, in general, split virus preparations induced substantially lower titres of HI antibody in mice than whole virus after one or two injections of the antigen.

Serologic and clinical response of children with sickle cell disease to bivalent influenza A split virus vaccine

INCREASED SUSCEPTIBILITY to severe bacterial infections is well documented in sickle cell disease? Although there is little evidence that non-bacterial infections pose similar problems, viral diseases such as influenza can predispose to infection with bacteria, particularly Streptococcuspneumoniae, which may be a major cause of death in children with SCD. 2' 3 In addition, viral infections may trigger a sickle cell crisis, necessitating hospitalization. 4 Prevention of viral diseases is therefore desirable. The availability of bivalent influenza vaccine (A/New Jersey/ 76-A/Victoria/75) during the latter half of 1976 provided an opportunity to evaluate the clinical response of young SCD patients to immunization.

The effect of ABO blood groups on the incidence of epidemic influenza and on the response to live attenuated and detergent split influenza virus vaccines.

The effect of blood group status on the incidence of epidemic influenza A (H3N2) infections and on serological response to influenza vaccination with killed subunit and live attenuated vaccines have been investigated during comparative vaccine trials in Western Australia. A significantly higher incidence of epidemic influenza was observed in subjects of blood group B compared with those of other blood groups, regardless of whether they had serological evidence of previous exposure to H3N2 antigens or not. Volunteers of different blood groups exhibited similar seroconversion frequencies to both vaccines after the administration of two doses, but a significantly higher proportion of blood group A subjects seroconverted after receiving their first dose of live attenuated vaccine compared with those of other blood groups. Although this finding was inconsistent with the increased incidence of epidemic influenza in subjects of blood group B, it is discussed in terms of the methods employed to obtain attenuation. Higher geometric mean HI antibody titres were observed in blood group O subjects after the administration of killed subunit vaccine. The results described in this report supported the contention that genetic factors linked to ABO blood groups may play a role in susceptibility to infection with influenza A virus, but that any association must be indirect.

Serologic Responses to Split and Whole Swine Influenza Virus Vaccines in Light of the Next Influenza Pandemic

dent Gerald Ford's emergency action to thwart wlhat appeared to constitute a threat of pandemic influenza. The events of the succeeding months have increasingly diminished the fear of pandemic spread and now justify thle belief tliat swine influenza at Fort Dix was a zoonosis that just didn't make tile grade in man. The epidemic at Fort Dix, while unusual for swine influernza virus, appeared to result from the conditions of the human population (crowding and stress) and not from thle virus itself, which bore all the discernible attributes of a virus of swine.

Evaluation of Commercially Prepared Vaccines for Experimentally Induced Type A/New Jersey/8/76 Influenza Virus Infections in Mice and Squirrel Monkeys

Mice and squirrel monkeys were vaccinated and subsequently challenged at selected times to evaluate the immunoprophylactic value of vaccines against influenza virus type A/New Jersey/76. Mice were challenged with virulent, homologous virus either 17 or 60 days after vaccination with 80 chick cell-agglutinating (CCA) units of whole-virus vaccine. Vaccinated mice showed minimal lesions and virus in lung tissue and had lower lung weights than unvaccinated controls. These mice had titers of hemagglutination-inhibiting (HAI) antibody in serum of greater than 1:400, but only traces of antibody were found in lung washes. Vaccinated squirrel monkeys had significantly less illness than unvaccinated controls when challenged with virulent virus 30 days after intramuscular immunization with 200 CCA units of whole virus or 400 CCA units of split virus given either once or twice (at 30-day intervals). Equal protection was observed in all monkeys despite the absence of serum HAI antibody in some monkeys after vaccination. Anamnestic reactions were observed only in monkeys vaccinated with whole virus. The possible roles of various immune factors and antibody to neuraminidase are discussed.

The serological response of experimental animals to inactivated whole and split influenza virus vaccines

The immune responses of mice, hamsters and ferrets were measured following immunization with varying doses of either inactivated, whole or split-adsorbed subunit influenza A/Port Chalmers/1/73 virus vaccine. In all animal model systems, the adsorbed subunit virus vaccine was more immunogenic in inducing serum HI and NI antibodies than whole virus vaccine. However, complete protection, as measured by the ability of the vaccines to inhibit viral replication, was apparent in the mouse and hamster systems but not in the ferret model. The merits of each animal as a model system for determining the immunogenicity of inactivated influenza vaccines in humans was assessed. Thus, in ferrets, 1200 i.u. of adsorbed subunit vaccine induced a mean serum HI antibody titre of 1:1163, whereas an equivalent dose of whole virus vaccine induced a mean titre of 1:166. In mice and hamsters, the highest doses of both vaccines induced high serum HI antibody titres; however, in mice 100 i.u. of split adsorbed vaccine promoted a mean serum HI antibody titre of 1:264 compared to a mean titre of 1:13 induced by a similar dose of whole virus vaccine. In hamsters, 25 and 5 i.u. of split adsorbed vaccine produced 6·6- and 10-fold higher serum HI antibody titres, respectively, than whole virus vaccine.

Vaccination of Pigs Against Hog Cholera (Classical Swine Fever) With a Detergent Split Vaccine

Four pigs were inoculated subcutaneously with a detergent (triton X 100) split hog cholera virus in Freund’s incomplete adjuvant. Four other pigs were in the same way inoculated with a detergent split bovine viral diarrhoea virus, also in Freund’s incomplete adjuvant. In the experiment were used 3 control pigs. The vaccinations were repeated after 3 weeks. All pigs were challenged with highly virulent hog cholera virus (Tubingen) 12 weeks after primary inoculations. Signs of hog cholera were only noted in the control pigs. This introductory experiment was succeeded by a larger experiment with subcutaneous inoculations of: 10 pigs with detergent split hog cholera virus in Freund’s incomplete adjuvant, 10 pigs with detergent split hog cholera virus in a saponin (Quil A) solution, 10 pigs with detergent split bovine viral diarrhoea virus in Freund’s incomplete adjuvant, 10 pigs with detergent split bovine viral diarrhoea virus in the Quil A solution plus 5 control pigs. The vaccinations were repeated after 3 weeks, and finally all pigs were challenged 9 weeks later with the highly virulent hog cholera virus strain. With the exception of 1 animal which died accidentally, all animals survived in the groups inoculated with the Quil A vaccines and in the group inoculated with the detergent split hog cholera virus/oil adjuvant vaccine. In the group inoculated with the detergent split bovine viral diarrhoea virus/oil adjuvant vaccine, some of the pigs died of hog cholera.

Comparative trials of live attenuated and detergent split influenza virus vaccines

Comparative clinical trials of live attenuated and detergent-split subunit influenza virus vaccines were undertaken with 1048 volunteers in Western Australia. Volunteers were divided into three main groups, each of which received either live virus vaccine or a saline control administered intranasally, or subunit vaccine injected subcutaneously. No differences were recorded between the three groups in their post-vaccination symptoms. Serum samples were collected at various times up to 50 weeks after vaccination, and antibody titres were measured by haemagglutination-inhibition (HI) tests and, for 231 volunteers, by virus neutralization tests. The two vaccines were almost equivalent in inducing seroconversion in vaccinees with pre-trial HI titres of 96 or less, but the subunit vaccine stimulated a higher geometric mean HI antibody titre. The longevity of the HI antibody response was greater for the live virus vaccine. The height of the response and the longevity of neutralizing antibody were the same for both vaccines. Both vaccines provided a high degree of protection against epidemic A/England/42/72 influenza, and some protection against A/Port Chalmers/1/73 influenza.

Humoral and secretory antibody responses to immunization with low and high dosage split influenza virus vaccine

Tri (n-butyl) phosphate (TNBP)-split vaccine containing 6400 CCA units of influenza virus A/Aichi/68 (H3N2) was given intramuscularly to a group of volunteers. The changes in serum haemagglutination-inhibiting (HI) and nasal wash neutralizing antibody were measured, and the results compared with that of volunteers given a TNBP-split vaccine containing 400 CCA of the same virus. More volunteers given the high-dose vaccine developed a fourfold rise in serum HI antibody, and there was a greater increase in geometric mean titre in this group. In addition, more volunteers given the high-dose vaccine developed detectable nasal wash neutralizing and anti-neuraminidase antibodies. Following low-dose vaccine, the production of nasal wash antibody was not related to the serum antibody response. For both groups given vaccine, antibody was detected most frequently in nasal washings with relatively high levels of protein and IgA; the concentration of IgA was also directly related to the protein concentration.

Towards a split influenza virus vaccine

A simple method is proposed whereby extracts of influenza virus can be injected intraperitoneally in mice and protect them against subsequent challenge by live virus by the intranasal route.

Protection With Split and Whole Virus Vaccines Against Influenza

In a trial of influenza virus vaccines, 3,103 students (13.6% of a campus population) were given a standard dose of vaccine (400 chick-cell agglutination [CCA] units of influenza A and 300 CCA units of influenza B) in October or December 1971. Fifty-eight percent of the vaccinees received a tri-(n-butyl) phosphate (TNBP) split virus vaccine; 42% received inactivated whole virus vaccine. In January, a discrete influenza outbreak occurred. Influenza A/HK virus (H3N2) was recovered as the predominant infecting agent. In a three-week period, 168 students sought medical assistance for febrile influenza. Only eight (4.8%) of these were vaccinees, representing a 69% reduction from the attack rate in nonvaccinees. The protection derived from split virus and whole virus vaccine was similar.

Protection with split and whole virus vaccines against influenza

Protection with split and whole virus vaccines against influenza

Humoral and cellular response in humans after immunization with influenza vaccine.

The peripheral blood lymphocyte response and hemagglutination inhibition antibody titers were measured in nine adults before and after immunization with a killed split influenza virus vaccine. Cord blood lymphocytes were tested with the influenza antigen to exclude a nonspecific mitogenic effect. All of the subjects demonstrated preexisting antibody titers and antigen recognition by lymphocytes prior to immunization. The in vitro lymphocyte response after vaccination parallels the humoral antibody response to influenza antigen.