Homologous and heterologous antibody responses to subunit influenza virus vaccine.

In a group of 32 adult volunteers given subunit influenza virus vaccine containing 250 international units (i.u.) of A/Victoria/3/75, 250 i.u. of A/Scotland/840/74 and 300 i.u. of B/Hong Kong/8/73, there were substantial increases in the geometric mean homologous haemagglutination-inhibiting (HI) antibody titres. There was also substantial boosting of the antibodies to the earlier variants of the Hong Kong (H3N2) series and to a later variant of the Asian (H2N2) series. There was no boosting of antibodies to the A/FM/1/47 strain, a representative member of the H1N1 series, but two individuals showed substantial rises to A/PR/8/34 (HON1). There were increases in the HI titre of antibodies cross reactive with two recent isolations, A/Texas/1/77, and A/Victoria/35/77, but the majority of vaccinees failed to reach antibody titres likely to be protective against such strains.     

Protection by a polyvalent influenza vaccine and persistence of homologous and heterologous HI antibodies during a period of two epidemic seasons.

A split-product influenza A vaccine which contained an influenza B strain (B/Hong Kong/8/73) and two influenza A strains, antigenically identical with A/Fort Dix/741/76 (Hsw1N1) and A/Victoria/3/75 (H3N2), was offered to personnel of the CPHL. Changes in the antibody status were followed with serum samples collected from 153 participants on the day of vaccination and 1, 13 and 18 months thereafter. During the two epidemic seasons in the trial period there were only four serological influenza A infections (2.6%) among the vaccinees. This is one eighth of the corresponding infection rate (22%) in the general population estimated on the basis of other indices. The vaccinees'' antibody response was strongly influenced by the age of the individual subjects. During the trial period the decrease in the antibody titres slowed down. The geometric mean titres of homologous HI antibodies were still substantially higher at the end of the period than at the beginning. This also applied to heterologous antibodies against H1N1 viruses in persons born between 1926 and 1952. In participants born after 1952, the vaccine was not able to evoke these antibodies, and in participants born in or before 1925 the boosting effect was poor.     

Recycling of H1N1 influenza A virus in man--a haemagglutinin antibody study.

Sera from people born between 1883 and 1930 and collected in 1977 were tested for the presence of HI antibodies to A/FM/1/47 (H1N1) virus and three recently (1977 and 1978) isolated influenza A-H1N1 viruses. The highest frequency of high-titred antibody to the four H1N1 viruses was detected in sera from people born in 1903-4, i.e. 42, 54, 38, and 22% had antibody against A/FM/1/47, A/Hong Kong/117/77, A/Brazil/11/78, and A/Fukushima/103/78 respectively. The birthdate groups 1896-1907 showed a higher percentage of HI antibody titres greater than or equal to 18, greater than or equal to 50, greater than or equal to 100 or greater than or equal to 1600 against the four H1N1 viruses than the birthdate groups 1907-30. This indicates the existence of an era, 1908-18, in which, apart from the H3N2 virus (1900-18), the H1N1 virus was epidemic among the human population.     

Boosting immunity to influenza H5N1 with MF59-adjuvanted H5N3 A/Duck/Singapore/97 vaccine in a primed human population

In 1997, influenza A/Hong Kong/97 (H5N1) emerged as a potential human threat. In 1999, a randomised study comparing two doses of MF59-adjuvanted and non-adjuvanted influenza A/Duck/Singapore/97 (H5N3) surface-antigen vaccine found non-adjuvanted vaccine was poorly immunogenic. Addition of MF59 significantly boosted antibody to H5N1 to levels associated with protection. At 16 months, we undertook a follow-up study to assess the effect of H5N3 revaccination. Geometric mean titres (GMTs) of antibody by haemagglutination-inhibition (HI), microneutralisation (MN) and single radial haemolysis (SRH) indicated that protective antibody titres did not exist at 16 months after two-dose priming. Twenty-one days after revaccination, there was significant boosting of antibody compared to GMTs achieved 21 days after two-dose priming in the original study (P<0.001). MF59 significantly increased GMTs of antibody when compared to non-adjuvanted vaccine (P<0.001).     

Changes in the antibody status of a population following epidemic infection by influenza virus A2/Hong Kong/1/68

The haemagglutinin of influenza virus A2/Hong Kong/1/68 was shown to be markedly different from that of previously isolated A2 virus strains. No haemagglutination-inhibiting (HI) antibody to A2/Hong Kong/1/68 virus was detected in serum specimens collected in 1966 from persons aged 60 years or less. In contrast, HI antibody tests with 270 sera collected in 1968 indicated that 9·6% had demonstrable HI antibody at low titres, and 35·2% of 454 postepidemic (1969) sera had demonstrable HI antibody at relatively high titres. Most sera from persons aged 80 years and more collected in 1968 and 1969 had demonstrable HI antibody to influenza virus A2/Hong Kong/1/68. No HI antibody to the Hong Kong virus was detected in pre-epidemic sera from children aged 6 months to 3 years, whereas 32% of postepidemic sera had HI antibody. The acquisition of HI antibody to A2/Hong Kong/1/68 was not accompanied by an increase in the incidence or titres of HI antibody to heterotypic A2 influenza viruses. For sera from children aged 4-11 years, an increase of HI titre to heterotypic A2 influenza was found.     

Strain specificity of serum antibody to the haemagglutinin of influenza A (H3N2) viruses in children following immunization or natural infection.

The specificity of serum anti-HA antibody from children immunized or infected with A/Victoria/75 (H3N2 or A/Texas/77 (H3N2) virus was examined using the single radial haemolysis test together with adsorption of antibody with three antigenic variants A/Hong Kong/68 (H3N2), A/Port Chalmers/73 (H3N2) and A/Victoria/75 (H3N2). The majority of young children reacted to vaccination or infection by producing strain-specific (SS) antibody to the homologous virus. A small proportion of children''s sera contained cross-reacting (CR) antibodies capable of reacting with the haemagglutinins of all antigenic variants of the sub-type including A/HK/1/68. In contrast, most adults reacted immunologically to either vaccination or infection by producing CR antibody, reacting with all variants of the antigenic subtype including the prototype virus A/HK/1/68 (H3N2).     

A comparison of live and inactivated influenza A (H1N1) virus vaccines. 2. Long-term immunity.

Groups of volunteers were immunized with one of three influenza virus vaccines, and the resistance to challenge infection with attenuated influenza A (H1N1) virus was measured 8 months later. The vaccines were aqueous subunit influenza A/USSR/77 (H1N1) vaccine, aqueous subunit influenza B/Hong Kong/73 vaccine, or attenuated influenza virus A (H1N1) vaccine. The B virus vaccine was included as a control to assess the incidence of natural A virus infection during the study period. A proportion of the B virus vaccinees had pre-existing A (H1N1) virus antibody and were used to study the immunity conferred by natural infection to the live virus challenge. The serum antibody responses were measured at 1 and 8 months after immunization. The results showed that all the vaccines induced serum HI antibody in a proportion of the volunteers; however, after 1 month, higher titres of serum antibody were found in volunteers given inactivated A vaccine than in those given live attenuated A virus vaccine. Eight months post-immunization the titres of serum antibody in volunteers given inactivated vaccine had declined significantly, but there were no changes in the antibody titres of those given live virus vaccine. The incidence of infection by the challenge virus at 8 months post-immunization was directly related to the serum antibody titres 1 month post-immunization; no evidence was obtained to suggest that those given live virus vaccine had a more solid immunity than those given inactivated vaccine.     

Use of the single radial diffusion technique for influenza antibody surveys

A survey of antibody to the influenza A/Hong Kong/1/68 (H3N2) and the variant A/England/42/72 (H3N2) was carried out by two methods, haemagglutination-inhibition (HI) and single radial diffusion (SRD). A comparison of the sensitivity of the two tests indicated that the proportion of sera that produced a zone of 2.5 mm or larger in the SRD tests was equivalent to the proportion that gave HI titres of 1: 40 or more. The test was carried out on serum samples collected from various regions of England. The results showed that approximately 70% had antibody to the A/HK/1/68 virus and only 35% had antibody to A/England/42/72. The SRD test was shown to have several practical advantages for seroepidemiological studies.     

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.     

Haemagglutination-inhibition antibodies against influenza A and influenza B in maternal and neonatal sera.

Haemagglutination inhibition (HI) antibodies against the influenza viruses A/Hong Kong/8/68 (H3N2) and B/Nederland/77/66 were determined in 420 paired sera from mothers and newborns (umbilical cord sera), sampled in 1970-1. A higher concentration of antibodies against influenza A virus was found more frequently in neonatal than in maternal sera. By contrast, low titres against influenza B virus were more frequently observed in neonatal than in maternal sera. Maternal age, duration of pregnancy, and birth-weight did not affect the results of the tests. It is suggested that the titre of the newborn against an epidemic influenza virus can be predicted from that of the mother. Furthermore, the maternal titre may be an indication of the susceptibility of the newborn infant to influenza infections.     

Haemagglutination-inhibition antibodies against influenza A and influenza B in maternal and neonatal sera

Haemagglutination inhibition (HI) antibodies against the influenza viruses A/Hong Kong/8/68 (H3N2) and B/Nederland/77/66 were determined in 420 paired sera from mothers and newborns (umbilical cord sera), sampled in 1970-1. A higher concentration of antibodies against influenza A virus was found more frequently in neonatal than in maternal sera. By contrast, low titres against influenza B virus were more frequently observed in neonatal than in maternal sera. Maternal age, duration of pregnancy, and birth-weight did not affect the results of the tests. It is suggested that the titre of the newborn against an epidemic influenza virus can be predicted from that of the mother. Furthermore, the maternal titre may be an indication of the susceptibility of the newborn infant to influenza infections.     

Immunity to influenza in ferrets: VII. Effect of previous infection with heterotypic and heterologous influenza viruses on the response of ferrets to inactivated influenza virus vaccines

Normal ferrets did not produce serum antibody following immunization with 200 i.u. of inactivated A/Hong Kong/68 influenza virus vaccine and were found to be susceptible to subsequent challenge infection with A/Hong Kong/68 virus. High titres of virus were recovered from nasal washings collected 3 days after infection, serum antibody was produced, increased nasal protein was detected and HI antibody was detected in nasal washings. Ferrets infected with influenza virus A/PR/8/34 7 weeks before immunization with inactivated A/HK/68 virus did, however, produce serum HI antibody to A/HK/68 virus. This antibody conferred partial immunity to challenge infection with A/HK/68 virus, as shown by decreased titres of virus in nasal washings and reduced levels of nasal protein. Previous infection of ferrets with influenza virus B/Ann Arbor/66 did not result in the production of serum antibody to A/HK/68 virus following immunization with A/HK/68 vaccine and the animals were completely susceptible to subsequent challenge infection with A/HK/68 virus. Differences in the amount of nasal protein and nasal antibody produced after A/HK/68 infection were also found in ferrets previously infected with either A/PR/8/34 or B/AA/66 virus, compared with normal ferrets.     

Cross-reactive antibodies in middle-aged and elderly volunteers after MF59-adjuvanted subunit trivalent influenza vaccine against B viruses of the B/Victoria or B/Yamagata lineages

This study evaluated whether MF59-adjuvanted subunit trivalent influenza vaccine for the 2003/04 winter season (A/Moscow/10/99, H3N2; A/New Caledonia/20/99, H1N1; B/Hong Kong/330/01) would confer protection against mismatched and frequently co-circulating variants of influenza B/Victoria- and B/Yamagata-like virus strains. Haemagglutination inhibiting (HI) antibodies were measured in middle-aged and elderly volunteers against the homologous B/Victoria-like vaccine strain (B/Hong Kong/330/01) and against mismatched B/Victoria-like (B/Malaysia/2506/04) and B/Yamagata-like (B/Singapore/379/99 and B/Shanghai/361/02) strains. Immunization induced significant increases in the amounts of HI antibodies against all influenza B strains under investigation. However, the responses against the heterologous B/Shanghai/361/02 virus did not reach the desirable values of seroprotection. An age-dependent decline of the responses was found for B/Victoria-like antigens, but not for B/Yamagata-like strains. Although further studies are needed, our data support the recommendation of including influenza B viruses of the B/Victoria and B/Yamagata lineages in the future influenza vaccine preparations.     

Hong Kong influenza in the Royal Air Force 1968-70

A prospective serological and clinical study of the epidemics due to the A2/Hong Kong/68 influenza virus was made during the winters 1968-9 and 1969-70 in volunteer subjects in the Royal Air Force. In October 1968 nearly all subjects had haemagglutination inhibiting (HI) antibody to the A2/Singapore/57 virus and more than half had antibody to strains more recently prevalent in Britain. The proportion with HI antibody to A2/Hong Kong/68 increased from 31% in October 1968 (most at low titres) to 44% after the first epidemic and 72% after the second (most at high titres). Serological infection rates were much lower in those who had detectable antibody at the beginning of each winter than in those who did not. Respiratory illnesses coupled with serological evidence of influenza infection during the winter were rare in persons with an initial titre of HI antibody of 1/40 or more. Infection in the first winter conferred complete protection against infection, with or without illness, in the second. In both epidemics about half those with serological evidence of infection had no reported illness.     

Seroprevalence of antibody to pandemic influenza A (H1N1) 2009 among healthcare workers after the first wave in Hong Kong

During the first wave of an influenza pandemic prior to the availability of an effective vaccine, healthcare workers (HCWs) may be at particular risk of infection with the novel influenza strain. We conducted a cross-sectional study of the prevalence of antibody to pandemic influenza A (H1N1) 2009 (pH1N1) among HCWs in Hong Kong in February-March 2010 following the first pandemic wave. Sera collected from HCWs were tested for antibody to pH1N1 influenza virus by viral neutralisation (VN). We assessed factors associated with higher antibody titres, and we compared antibody titres in HCWs with those in a separate community study. In total we enrolled 703 HCWs. Among 599 HCWs who did not report receipt of pH1N1 vaccine, 12% had antibody titre ≥1:40 by VN. There were no significant differences in the age-specific proportions of unvaccinated HCWs with antibody titre ≥1:40 compared with the general community following the first wave of pH1N1. Under good adherence to infection control guidelines, potential occupational exposures in the hospital setting did not appear to be associated with any substantial excess risk of pH1N1 infection in HCWs. Most HCWs had low antibody titres following the first pandemic wave.     

Comparison of haemagglutination-inhibition and single radial haemolysis techniques for detecting antibodies to influenza A and B viruses

The sensitivities of haemagglutination-inhibition (HI) and single radial haemolysis (SRH) techniques in detecting antibodies against influenza A/Bangkok/1/79, A/Brazil/11/78, B/Singapore/222/79, B/Hong Kong/5/72 strains, in human sera were compared. For antibodies to influenza B viruses the HI tests employing ether-treated antigens were also evaluated. The SRH technique appears to be more sensitive for detecting protective titres of antibodies against influenza B strains and influenza A/Brazil/11/78.     

Comparison of haemagglutination-inhibition and single radial haemolysis techniques for detecting antibodies to influenza A and B viruses.

The sensitivities of haemagglutination-inhibition (HI) and single radial haemolysis (SRH) techniques in detecting antibodies against influenza A/Bangkok/1/79, A/Brazil/11/78, B/Singapore/222/79, B/Hong Kong/5/72 strains, in human sera were compared. For antibodies to influenza B viruses the HI tests employing ether-treated antigens were also evaluated. The SRH technique appears to be more sensitive for detecting protective titres of antibodies against influenza B strains and influenza A/Brazil/11/78.     

Surveillance of influenza in Houston, Texas, USA: gradual transition from A/Victoria/75 (H3N2) to A/Texas/77 (H3N2) predominance and antigenic characterization of "intermediate" strains

Influenza epidemics in Houston, Texas, USA, during the winters of 1975-76, 1976-77, and 1977-78 were attributed to A/Victoria/3/75 (H3N2), B/Hong Kong/5/72, and A/Texas/1/77 (H3N2)-like viruses, respectively. Both A/Victoria and A/Texas viruses were detected towards the end of the 1976-77 epidemic and throughout the 1977-78 epidemic. To determine if there had been a gradual transition in the predominant strain, 267 viral isolates from the 1975-76 epidemic were tested for A/Texas virus. Eight specimens (3%) that appeared to contain A/Texas antigens were cloned and retested with specific antisera prepared in guinea-pigs and ferrets. One virus was identical to A/Texas/1/77 virus, two reacted like A/Victoria/3/75 and five reacted equally well with antisera prepared against A/Victoria/3/75 and A/Texas/1/77 viruses (bridging strains). The six viral isolates containing A/Texas antigens were obtained at different times during the epidemic, from all parts of the city, from males and females aged between 1 and 20 years.     

Comparison of the protective efficacies of the live vaccine RIT 4025 and an inactivated vaccine against a natural heterologous A/Victoria/3/75 infection.

A clinical trial was initiated in South Africa before the winter season of 1976. The study involved 253 volunteers divided into three groups of vaccinees and one control group. Two groups of vaccinees were inoculated with either one or two doses at 2 weeks' interval (10(7.2) EID 50/dose) of the RIT 4025 live recombinant strain [A/Scotland/840/74 (H3N2) serotype] and one group received one injection of an inactivated vaccine [A/Port Chalmers/1/73 (H3N2), 360 i.u., A/Scotland/840/74 (H3N2), 300 i.u. and B/Hong Kong/8/73, 300 i.u./dose]. The serum antihaemagglutinin antibody responses against the heterologous A/Victoria/3/75 strain as measured by the single radial haemolysis test were satisfactory and not statistically different in all groups of vaccinees. On the other hand, the antineuraminidase antibody response was better in the group receiving the killed vaccine. At the end of the influenza season, A/Victoria/3/75 infections were confirmed serologically. Only 12% of the infections were symptomatic. The infection rate was significantly reduced in the live vaccine groups, whereas in the killed vaccine group the percentage of infection was lower but not significantly different from that in the placebo group.     

Interpretation of responses and protective levels of antibody against attenuated influenza A viruses using single radial haemolysis.

Antibody determinations against H3N2 and H1N1 type A influenza viruses were carried out on paired sera obtained from volunteers taking part in influenza virus vaccine studies, using both the haemagglutination-inhibition (HI) and single radial haemolysis (SRH) test. Good correlation between the HI and SRH test was found for both H3N2 and H1N1 antibody and the zone area increases corresponding to significant SRH antibody rises determined for both virus strains. In both H3N2 and H1N1 vaccine studies, intranasal infection of the volunteers with live attenuated viruses was involved and by the measurement of HI and SRH antibodies prior to and following infection, levels of antibody equating with protection against the infecting viruses could be estimated. For the HI test the antibody titres associated with 50% protection were 42 for H1N1, and 44 for H3N2 viruses; for the SRH test, 50% protection was associated with zone areas of 20.0-25.0 mm2 for both H1N1 and H3N2 viruses.