Effects of repeated annual influenza vaccination on antibody responses against unchanged vaccine antigens in elderly frail institutionalized volunteers.

BACKGROUND: Concern about the possibility that annually repeated influenza immunizationmayinduce a lower antibody response than first vaccination. OBJECTIVE: To ascertain the cumulative effects of yearly vaccination on serological response to unaltered vaccine antigens in the elderly. METHODS: The haemagglutination-inhibiting (HI) antibody response was examined in 158 elderly institutionalized frail volunteers subdivided in 3 groups according to the sequential winters in which each subject received a trivalent inactivated influenza vaccine. The study, conducted over 5 consecutive winters (from 1998/99 to 2002/03), reports the antibody response only for sequential years (2 or 3) in which the vaccine strain examined was not altered. RESULTS: Significant increases in the values of HI antibody titres were observed after vaccination in each year examined against the different influenza vaccine strains used, except against B antigen in the second of the 3 winters studied (1999/00). The antibody responses found were not always adequate, i.e. at levels above the currently requested values for commercial vaccines (post-vaccination seroprotection rate >/=1:40, increases in geometric mean titres >/=2, positive responses >/=30% compared with pre-vaccination), probably because of old age (mean age >/=81 years) and the presence of underlying diseases in a high percentage of volunteers (>/=86%). The most frequent chronic diseases found werecardiovascular diseases (48%), endocrine disorders (19%), functional disability (10%) and pulmonary diseases (4%). The post-vaccination values observed in the sequential years were in general similar for A/H3N2 and A/H1N1 vaccine strains. A decrease, however, for some parameters at statistically significant levels, was observed against B antigen following repeated vaccine administrations. CONCLUSION: Our data seem to support the possibility of a slight impairment of HI antibody response against unaltered influenza vaccine antigens, especially for influenza strains that have circulated for prolonged periods of time. Indeed a tendency to a lower response was found only against B/Beijing antigen, introduced in the vaccine composition in the winter 1995/96, but not against the A/H3N2 and A/H1N1 vaccine strains, which weremore frequently changed.

A viral immunodepressive factor associated with experimental mouse tumors.

An immunodepressive factor (IDF) capable of inhibiting the rejection of allogeneic lymphomas in mice was detected in a number of transplantable mouse tumors. The allograft reactivity of the host was impaired for at least 10 months after IDF administration. No clinical evidence of bacterial or viral infection was detected in IDF-treated animals. Sera collected from mice at various intervals after injection of IDF showed a great increase in the IDF-titer. The IDF persisted in mouse sera for at least 3 months, whereas no IDF activity was found in sera of rats given injections of the factor. IDF was capable of replicating in vitro in mouse embryo cells, but not in rat embryo or HeLa cell cultures. IDF was inactivated in vitro by heat (65 degrees for 30 min), ultraviolet light or ether, but not by ethyl alcohol. These studies indicate that IDF is a virus capable of producing a long-lasting asymptomatic infection specifically interfering with the host's allograft reactivity. In several instances a close association between the lactic dehydrogenase virus and IDF was found. Nevertheless no conclusion was reached on the identity or nonidentity between the two viruses.

Inhibition of hybrid resistance to lymphomas by inactivated tumor cells in lethally irradiated mice.

Two murine lymphomas, L5MF-22 of B10.129(5M) (H-2b) origin and P388 of DBA/2 (H-2d) origin, were inoculated into lethally irradiated hybrid (C57BL/6 x DBA/2)F1 (B6D2F1) mice (H-2b/H-2d). Marked localized graft resistance was found in the spleen and occasionally in the liver of recipient mice as a result of a non-T-dependent hybrid resistance (HR). Significant reduction of HR of B6D2F1 mice could be obtained when viable lymphoma cells were inoculated along with inactivated cells of the same tumor or of genetically related leukemias. These results suggest that in vivo competition for HR effectors can take place in mouse spleen and liver leading to a depression of the localized resistance.

Effect of in vitro interferon treatment on the rapid clearance of murine leukemia cells in vivo.

Previous work showed that interferon (IFN) can protect target cells from NK mediated lysis in vitro. In the present study we investigate the effect of IFN alpha/beta or IFN gamma treatment of three different murine leukemia cell lines. For this purpose FLC-745 (susceptible to the antiproliferative activity of IFN alpha/beta and gamma), FLC-3C18 (IFN alpha/beta -resistant and IFN gamma - susceptible) of DBA/2 origin and EL-4 (IFN alpha/beta - susceptible and IFN gamma - resistant) leukemia of C57B1/6 origin were treated with IFN alpha/beta or gamma in vitro and assayed for their susceptibility to natural resistance measured in vivo as organ rapid clearance 4 hr after iv injection into syngeneic mice. Using young or Poly I:C stimulated hosts, but not mice with low levels of natural resistance (i.e. older animals or mice treated with cyclophosphamide), slower elimination of treated cells was observed with: (a) FLC-745 cells treated with IFN alpha/beta and IFN gamma and (b) FLC 3C18 treated with IFN gamma. Such a delayed clearance was not observed with: (a) FLC-3C18 cells treated with IFN alpha/beta and (b) EL-4 leukemia cells preincubated with IFN alpha/beta or IFN gamma. These results suggest that under selected conditions IFNs can protect leukemic cells from in vivo natural reactivity.

Retinol dietary intake and oral leukoplakia development.

To investigate the clinical outcome of unbalanced diet in patients with oral precancerosis and to assess a possible relationship between dietary factors and the development of oral leukoplakia, a case-control study was carried out within a cohort of 53 subjects treated at our Centre in October-November 1997. Enrolled subjects and suitable controls underwent a careful interview on their own alimentary habits with a particular interest in retinol and carotenoids major sources. An individual qualitative and quantitative assessment of retinol-equivalents dietary intake, yielding average values for each group, allowed to compare the cohorts and to relate data also to tobacco use and to the severity of histopathological findings. Case levels were always significantly lower than controls (P<0.001), disregarding smoking, whilst no difference resulted between smokers and non smokers within the same groups. No statistical influence seemed to link alimentary vitamin A to the development of oral dysplasia but this work strengthen the epidemiological opinion that specific dietary factors are of great importance in oral oncology.

Flow cytometric measurement of intracellular IFN-gamma induction in aged subjects before and after parenteral influenza vaccination.

Flow cytometric analysis, used to study intracellular expression of IFN-gamma in peripheral blood mononuclear cells (PBMC) from aged volunteers before and after parenteral influenza vaccination, was found capable of rapidly detecting influenza antigen induced variation of IFN-gamma expression. Although the vaccine was capable of generating a satisfactory antibody response, it did not stimulate an increase in the percentage of IFN-gamma positive cells.

Immunobiological aspects of the nude mouse model relative to human cancer chemosensitivity tests.

The nude mouse model (NMM) has been proposed for testing chemosensitivity of human cancer cells with encouraging results. However, nude mice cannot be considered as unreactive "test tube" recipients of allogeneic or xenogeneic tissues, since a variety of immunological functions are fully represented in these hosts. In general, graft resistance against tumour cells can be classified as: (a) elicitable responses (ER), thymus-dependent, evoked by tumour-associated antigens; (b) natural resistance (NR), T-independent, not requiring previous exposure to transplantation antigens. Graft resistance may therefore play a substantial role even in the absence of a functional T-cell system. This has been demonstrated in irradiated euthymic mice capable of rejecting Hh-incompatible lymphomas, and in nude mice, where NR-type responses are particularly efficient, either in vitro or in vivo. It is reasonable to assume that these responses could interfere with tumour responsiveness to the chemotherapeutic effects of drugs. In fact, marked synergistic effects have been observed in conventional hosts combining chemotherapy with limited ER, or with Hh-type NR present in lethally irradiated mice. Similar combined effects could occur in nude mice bearing NR-susceptible human tumours in the course of chemosensitivity assays. These considerations have led to the search for privileged sites associated with low levels of NR; previous results show that these responses appear to be substantially absent in the brains of conventional mice and extremely low in the same organ of congenitally athymic hosts. It follows that the brain of the nude mouse appears to be a suitable site for human tumour cell growth, unaffected by or minimally subjected to NR.

An influenza B outbreak during the 2007/2008 winter among appropriately immunized elderly people living in a nursing home.

The study evaluated the immunogenicity and efficacy of a trivalent subunit MF59-adjuvanted influenza vaccine (A/Wisconsin/67/05 (H3N2), A/Solomon Islands/3/06 (H1N1) and B/Malaysia/2506/04) in preventing serologically diagnosed infections in a group of 67 institutionalized elderly volunteers during 2007/2008 winter, characterized by co-circulation of drifted A/H3N2, A/H1N1 and B influenza viruses. Influenza vaccination induced a significant increase in the amounts of hemagglutination inhibiting antibodies, both against the vaccine and the epidemic drifted strains. However, vaccination did not prevent the circulation of the new drifted influenza B virus (B/Florida/4/06-like), belonging to the B/Yamagata/16/88-lineage, antigenically and genetically distinct from B/Victoria/2/87-lineage viruses from which the vaccine B strain was derived.

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.

Influenza vaccination in patients on long-term anticoagulant therapy.

The study evaluates the risk/benefit of influenza vaccination in patients on stable long-term oral anticoagulant therapy (OAT). One hundred and four consecutive patients with indication for influenza vaccination were randomized to receive one dose of 2004/2005 influenza vaccine followed by placebo after 6 weeks, or vice versa, in a cross-over blinded trial. All patients were tested for anticoagulation levels and for hemagglutination inhibiting antibody titres against the influenza vaccine antigens. The highly protective antibody titres induced by influenza vaccination and the absence of statistically relevant interactions between vaccination and OAT suggest that influenza vaccination can be used safely and successfully in elderly patients on OAT.

An influenza A/H3 outbreak during the 2004/2005 winter in elderly vaccinated people living in a nursing home.

This study examined the antibody response against the three vaccine antigens and the epidemic A/H3N2 drift variant (A/California) and the prevention of laboratory diagnosed influenza infections in a group of elderly institutionalized people vaccinated with the 2004/2005 influenza vaccine. Antibody titres were measured by hemagglutination inhibition (HI) in sera collected before and 1 month after vaccination. Laboratory diagnosis was done examining throat swabs (RT-PCR or MDCK cell culture) or by serology (seroconversion comparing HI titres in sera collected 1 and 5 months after vaccination). Results obtained showed that influenza vaccination induced an adequate immune response against the three vaccine antigens and the epidemic A/H3N2 variant, however it was not capable of preventing an influenza outbreak due to the new A/H3N2 (A/California) variant.

[Influenza in Umbria from September 1977 to May 1978]. / Indagini sulla influenza in Umbria nel periods Settembre 1977 - Maggio 1978.

From September 1977 through May 1978 a research project was conducted to determine the presence of Haemagglutination Inhibiting Antibodies (H.I.A.) on three influenza viruses: A/Victoria/3/75, A/URSS/90/77 and B/Hong Kong/8/73. Participating in the project were 1066 persons residing in the region of Umbria, Italy, ages ranging from 0- - 60. The results obtained showed the possibility of diffusion in Umbria of all three of the strains studied due to the low protective levels found in the participating subjects. In fact, the protective levels (> 1:40) are present only in a small percentage of subjects (31% protected against A/Victoria/3/75; 1% protected against A/URSS/90/77 and 10% protected against B/Honk Kong/8/73). An influenza strain of type A, related to A/URSS/90/77 was isolated in Perugia (Umbria) in March of 1978.

Natural resistance of mice pretreated with 5-(3,3'-dimethyl-1-triazeno)-imidazole-4-carboxamide (DTIC) + cyclophosphamide (Cy) against virus-induced lymphoma cells.

Virus-induced leukemia was inoculated into histocompatible or allogeneic hosts pretreated with 5-(3,3'-dimethyl-1-triazeno)-imidazole-4-carboxamide (DTIC) + Cyclophosphamide (Cy), which abrogate endogenous cell proliferation and T-dependent graft responses, but not selected "natural resistance" (NR) activities. Marked impairment of lymphoma cell growth occurred mainly in the spleen of allogeneic mice with respect to that of histocompatible controls. Tumor inhibition was still present when lymphoma challenge was performed on day + 3 after Cy administration. Parallel studies on "natural killer" (NK) activity in vitro or in vivo showed that complete abrogation of the NK function was detectable on day + 3 or + 6 after Cy treatment. It was concluded that in vivo inhibition of lymphoma growth in mice pretreated with DTIC + Cy could be a drug-resistant NR at least in part distinguishable from the NK function.

[Antigenic characteristics of hamster cells transformed by polyoma virus]. / Caratteristiche antigeniche di cellule di criceto trasformate da virus polioma

A hamster cell line derived from a polyoma virus induced tumour (G-Py) was found positive for TSTA and S antigen detected by the surface immunofluorescence technique. Lung metastases were observed in some hamster able to reject a G-Py cell challenge dose injected by intradermal route.

Possible correlation between low antigenic drift of A(H1N1) influenza viruses and induction of HI antibodies.

This study examined whether, during a seven-year period of low A(H1N1) influenza virus antigenic drift (1988-1989 and 1994-1995, winters), humoral antibody response of elderly volunteers to influenza vaccines could suggest a lack of antibody pressure for drift. In all the years studied A/Taiwan/1/86, the A(H1N1) vaccine component, had a low ability to induce protective hemagglutination-inhibiting (HI) antibody titres (> or = 1:40). However a similar low immunogenicity was found for some of the different A(H3N2) strain variants of influenza virus, co-circulating in the same period and showing a regular extent of antigenic variations. Although our data could be at least in part explained by the type of study population (elderly and repeatedly vaccinated), postepidemic serological studies did not evidence a consistently lower ability in mounting protective immune response in elderly people as compared with younger against the influenza strains studied. Therefore, our present results did not exclude a true low immunogenicity of A/Taiwan and of some A(H3N2) influenza strains, circulating in the winters examined. This suggests that, besides the necessity to evade prior immunity, additional factors could influence the frequency of influenza viruses antigenic drifts.

Immunogenicity of trivalent subunit and split influenza vaccines (1989-90 winter season) in volunteers of different groups of age.

Trivalent split or subunit influenza vaccines [A/Shangai/11/87 (H3N2), A/Singapore/6/86 (H1N1) and B/Yamagata/16/88] recommended for the 1989-90 winter season and licensed in Italy, were administered to 149 volunteers of three different age groups (elderly, middle-aged and young). Antibody production was determined in pre- and postvaccination sera by haemagglutinin inhibition test and the results were evaluated as protection and response rates. The split vaccine was more immunogenic than the subunit preparation, especially against the B virus strain. Age had no obvious impact on the degree of responsiveness to vaccination.

Immune response to trivalent inactivated influenza vaccine in young and elderly subjects.

The antibody response (determined using the single radial haemolysis in gel technique) to inactivated whole-virion trivalent influenza vaccine [A/Leningrad/360/86(H3N2), A/Taiwan/5/87 and B/Ann Arbor/1/86], recommended for the 1987-88 winter season in Italy, in 49 elderly (age greater than or equal to 60 years) subjects was compared with the response in 23 young adult (age less than 60 years) volunteers. The subjects were prevalently healthy and a high percentage of young and old people had been repeatedly immunized against influenza in previous years. No significant differences were detected among age groups; moreover, the immune response measured by seroconversion or by a significant rise in antibody titre was constantly low.

In vivo natural antitumor resistance against murine EL-4 lymphoma cells in lethally irradiated syngeneic C57Bl/6 mice.

Natural resistance has been detected in lethally irradiated C57Bl/6 (B6) mice inoculated intravenously with the ascites form of a syngeneic B6 leukemia. EL-4 cells were injected into lethally irradiated (800 R) B6 mice and tumor cell proliferation was evaluated by 125IUdR uptake in different organs 4 days after the challenge. Differential growth of lymphoma cells was observed when young mice were injected as compared with older mice and when mice were treated with agents known to interfere with natural resistance (e.g., poly(I:C), FLV-P, carrageenan, cyclophosphamide, high doses of irradiated cells). Similar results were obtained by measuring rapid clearance of 125IUdR-labeled EL-4 cells from lungs of intact B6 mice. In vivo cold competition studies, employing EL-4 and several other tumor lines of the same or different haplotype, showed that only EL-4 and RBL-5 cells were capable of inhibiting syngeneic resistance against EL-4 tumor. On the contrary, YAC-1 lymphoma cells, the most susceptible target to natural killer-mediated cytotoxicity in vitro, did not compete. These results suggest that EL-4 cells express membrane determinants not detectable on normal H-2b parental bone marrow cells and are susceptible to natural resistance against hemopoietic tumor cells in lethally irradiated syngeneic B6 mice.

Natural resistance against hematopoietic cells in lethally-irradiated mice infected with Friend leukemia virus.

The influence of in vivo infection with the polycythemic substrain of Friend leukemia virus on noninducible ('natural') resistance against allogeneic normal or malignant grafts was studied in lethally irradiated mice. Parallel studies were performed on the NK system in the same experimental conditions. The results indicate that FLV-P infection of mice with full (DBA/2) vs partial (BALB/c and CD2F1) susceptibility did not suppress their in vivo natural resistance against bone marrow or El-4 leukemia cells. On the other hand, a decline in NK activity paralleled the progression of leukemic disease in the more susceptible DBA/2 hosts.

Immunization of elderly volunteers with the 1988-89 inactivated whole influenza vaccine: assessment of antibody responses by haemagglutination inhibition and single radial haemolysis tests.

The immunogenicity of inactivated whole trivalent influenza vaccines (A/Taiwan/1/86 (H1N1), A/Sichuan/2/87 (H3N2), and B ijing/1/87) recommended for the 1988-89 winter season was evaluated in 236 elderly (mean age 71 years) high risk volunteers. An overall significant increase in the number of subjects with protective haemagglutination inhibiting (HI) antibodies (titer > 1:40) against vaccine components was observed after vaccination. Nevertheless, a percentage of individuals (ranging from 56% to 62%) remained without protective antibodies and the number of people showing a positive response was limited (from 32% to 41%). By the comparative analysis of the results obtained examining the presence of protective levels of antibody in the sera from 91 volunteers using HI versus the single radial haemolysis (SRH) test, we obtained evidence for a higher sensitivity of SRH technique especially against B antigen.