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PubMed2006

Variations of Neospora caninum antibody levels in milk during lactation in dairy cows.

A longitudinal study was performed to investigate the variations of Neospora caninum antibody levels in individual milk during lactation as well as the association between antibody levels in serum and milk. Serum and milk samples of 15 milking cows were collected between February 2003 and September 2004 in three smallholder dairy farms in Khon Kaen province in northeast Thailand. All samples were analyzed for presence of antibodies by an N. caninum iscom ELISA test kit and the results were given as percent positivity (PP). The effects of time between calving and sampling, lactation number, and season on milk and serum PP were studied using Generalized Estimation Equations methods. All cows were antibody positive in either milk or serum at the first two consecutive samplings. Although serum and milk PP varied considerably, milk PP was consistently positive throughout the study. Cows of all lactation groups had a higher adjusted mean of milk PP at calving compared to later months after calving although the only significant difference was in first lactation. Serum and milk PP were always lower in first lactation than in second and later lactations. An adjusted mean of milk PP for cows classified as having serum PP> or =55 was significantly (P<0.05) higher than that of cows classified as having lower serum PP. Our results indicate that individual milk can be an alternative material to demonstrate presence of N. caninum antibodies in lactating cows.

Animals↗
PubMed2006

Prevalence of antibodies against Toxoplasma gondii and Neospora caninum in Hungarian red foxes (Vulpes vulpes).

In the present study we have investigated the seroprevalence to the protozoan parasites Toxoplasma gondii and Neospora caninum in 337 red foxes (Vulpes vulpes) from 16 out of 19 counties in Hungary. The foxes were originally collected within a National vaccination program against rabies. Antibodies to T. gondii were detected in as many as 228 (68%) of the foxes using a commercial direct agglutination test (DAT). In an indirect iscom ELISA, five foxes (1.5%) were positive for antibodies against N. caninum. The high prevalence of foxes positive for T. gondii might be explained by the widespread occurrence of the parasite in the diet of foxes. As a contrast, latent infections of N. caninum among red foxes in Hungary are much less common.

Agglutination Tests↗
PubMed2003

Recent advances in veterinary vaccine adjuvants.

Next generation veterinary vaccines are going to mainly comprise of either subunit or inactivated bacteria/viruses. These vaccines would require optimal adjuvants and delivery systems to accord long-term protection from infectious diseases in animals. There is an urgent need for the development of new and improved veterinary and human vaccine adjuvants. Adjuvants can be broadly divided into two classes, based on their principal mechanisms of action: vaccine delivery systems and 'immunostimulatory adjuvants'. Vaccine delivery systems are generally particulate e.g. emulsions, microparticles, ISCOMS and liposomes, and mainly function to target associated antigens into antigen presenting cells (APC). In contrast, immunostimulatory adjuvants are predominantly derived from pathogens and often represent pathogen associated molecular patterns, e.g. LPS, MPL and CpG DNA, which activate cells of the innate immune system. Recent progress in innate immunity is beginning to yield insight into the initiation of immune responses and the ways in which immunostimulatory adjuvants might enhance this process in animals and humans alike.

Adjuvants, Immunologic↗
PubMed1999

Duration of immunity, efficacy and safety in sheep of a recombinant Taenia ovis vaccine formulated with saponin or selected adjuvants.

The efficacy and safety of a recombinant Taenia ovis protein was tested in sheep using 13 different adjuvant formulations, including oil adjuvants, aluminium salts, saponin, Iscoms and DEAE-dextran. The oil adjuvants, saponin and DEAE-dextran gave the highest antibody responses and greatest degree of protection against challenge infection with T. ovis eggs. Duration of immunity studies with a saponin based vaccine showed that highly significant protection (>90% reduction of cyst numbers) was achieved when sheep were challenge infected one month after immunisation. Significant protection (79%) was still present when sheep were challenged 6 months after immunisation. The optimum dose for this batch of saponin was 10 mg, which stimulated a peak antibody titre of 38,400, 4 weeks after immunisation and did not cause injection site reactions. Dialysed saponin was shown to retain its adjuvant properties and allowed an increase in dose to 30 mg without site reaction, resulting in a peak antibody titre of 51,200.

Adjuvants, Immunologic↗
PubMed1999

HIV-1 vaccine-induced immune responses which correlate with protection from SHIV infection: compiled preclinical efficacy data from trials with ten different HIV-1 vaccine candidates.

The specific immune mechanisms necessary and/or sufficient to elicit HIV-vaccine protection remain undefined. Utilising the SHIV rhesus macaque model the immunogenicity as well as the efficacy of ten different HIV-1 vaccine candidates was evaluated. Comparison of the immune responses induced, with the ability of the vaccine to protect from SHIV infection provided a means to determine which type of immune responses were necessary for protection. Vaccine candidates included VLPs, DNA, subunit protein with novel adjuvant formulations, ISCOMs and pox-virus vectors. Protection from SHIV infection was achieved in approximately half of the animals which received a primary intravenous cell-free challenge. The presence of CTL in the absence of other effector responses did not correlate with protection from this route and type of challenge. Virus neutralising antibodies (Nab) appeared to be necessary but alone were insufficient for protection. If Ag-specific IFN-gamma and/or IL-4 as well as lymphoproliferative (LP) responses were found with the lack of a detectable IL-2 response, then protection was not observed. Immunity correlated with the magnitude of Nab responses, beta-chemokines and as well as balanced, qualitative T-helper responses.

AIDS Vaccines↗
PubMed1999

Vaccine delivery to animals.

For many years vaccination of animals has been practiced to prevent infectious diseases using inactivated organisms or modified live organisms. The live vaccines were effective but lacked safety. The vaccines made with inactivated organisms required an adjuvant to induce an immune response that was not as effective as either the clinical disease or live vaccines. An 'ideal' vaccine would induce effective immunity specific for the type of infection, have long duration, require minimal or no boosters, have impeccable safety, would not induce adverse reactions, and be easy to administer. The desire to meet these criteria, and especially safety, has resulted in the development of vaccines that do not depend on the use of the viable disease agent. The emphasis on subunit or inactivated vaccines that meet the desired criteria of a perfect vaccine has resulted in a critical need for better adjuvants and delivery systems. This has resulted in a technological innovation revolution with development of a wide array of different technologies to generate effective vaccines. This review will describe the historical relevance of adjuvants used for parenterally administered inactivated/subunit vaccines as well as describe some of the exciting technological advances including adjuvants (ISCOMS), delivery systems (recombinant vectors, microparticles), and novel approaches (transgenic plants, naked DNA) that are currently being, or will be used in the future, in the search for better, more effective vaccines that meet the current and future needs of veterinary medicine.

Journal Article↗
PubMed1992

Vaccination against acute respiratory virus infections and measles in man.

Several viruses may cause more or less severe acute respiratory infections in man, some of which are followed by systemic infection. Only for influenza and measles are licensed vaccines available at present. The protection induced by influenza vaccines, which are based on inactivated whole virus or viral subunits, depends largely on the matching of vaccine strain and circulating virus. Measles vaccines, which are based on attenuated live virus, have been quite effective in controlling the disease in vaccinated populations in the industrialized world. In developing countries, severe measles infections occur in infants from six to nine months of age, which necessitates the vaccination of children of less than six months. At that time maternal antibodies, that may interfere with the induction of protection, may still be present. Therefore, instead of using the parenteral route, the possibility to use the mucosal route of primary immunization is also investigated for vaccination with attenuated live measles vaccines. The use of inactivated measles vaccines has resulted in a state of immunity which upon exposure to the virus may induce an atypical measles syndrome including a severe pneumonia. Measles virus proteins presented in an iscom matrix have recently been shown to induce functional B and T cell responses to both the surface glycoproteins of the virus. These responses could also be induced in the presence of virus neutralizing antibodies and they proved to be protective in several animal model systems. Many of the problems that have been encountered in the development of measles vaccines, proved to be similar in the development of vaccines against other paramyxoviruses causing acute respiratory infections in man, including respiratory syncytial virus. Parenteral application of inactivated and attenuated live vaccines against these paramyxoviruses has generally had little success. Topical application of attenuated live vaccines has been more successful, and also the use of vaccinia recombinant viruses expressing foreign paramyxoviral glycoproteins has shown promising results in laboratory animals. Live vaccines based on adenovirus types 4 and 7 in oral enteric-coated vaccines, which lead to virus replication in the intestines but not in the respiratory tract have been included in military vaccination programs. The possibility to replace e.g. the E3 region with foreign DNA makes adenoviruses also suitable as cloning vectors for proteins of other respiratory viruses. Although live attenuated vaccines against some of the serotypes of rhinoviruses have shown promising results, the generation of a multivalent vaccine against this epidemiologically most significant cause of acute respiratory infections will be almost impossible, due to the multiplicity of serotypes involved.(ABSTRACT TRUNCATED AT 400 WORDS)

Humans↗
PubMed2003

Influence of maternal immunity on antibody and T-cell response in mice.

The prevalence of maternal antibodies (Abs) and an immature neonate immune system, which is inclined to a T-helper 2 (Th2) response, are factors that counteract active immunization in early life. In a mouse model, the maternal influence on an active immunization of 2-day-old offspring with Sendai virus (SV) envelope proteins was explored. Maternal immunizations were conducted with the same SV antigen preparation as used for offspring immunization, presented in three different formulations, namely micelles of SV (SV-MIC), Al(OH)(3)-adjuvanted SV-MIC (SV-aluMIC) for Th2 and immune stimulation complex (iscom)-adjuvanted SV (SV-ISC) for Th1. An inversely correlation was found between the immunoglobulin G2a (IgG2a) Abs of the mothers and the interleukin 5 (IL-5) levels of the offspring. Although a maternally derived immunity induced by SV-aluMIC suppressed both B- and T-cell responses of the newborn to SV-ISC immunization, significant priming effects of the neonatal immunization on IgG2a Abs and IFN-gamma levels were recorded after reimmunization at adult age.

Animals↗
PubMed2003

Immunogenicity and efficacy of recombinant subunit vaccines against phocid herpesvirus type 1.

Phocid herpesvirus type 1 (PhHV-1) is an alpha-herpesvirus that causes significant morbidity and mortality among young and immunocompromised harbour seals (Phoca vitulina) and therefore represents a major problem for seal rehabilitation centres. Consequently, there is a need for a safe and effective PhHV-1 vaccine. We tested an ISCOM-based recombinant PhHV-1 gB vaccine alone (gB) or with the addition of recombinant PhHV-1 gD (gBD) for (i). immunogenicity and protective efficacy against feline herpesvirus (FHV) infection in cats and (ii). their immunogenicity in seals. The FHV-cat model was chosen based on the close antigenic relationship between PhHV-1 and FHV. Upon challenge, all vaccinated (gB and gBD) cats excreted significantly less FHV (P<0.01) and gBD vaccinated cats showed less weight loss (P=0.05) than the mock-vaccinated cats. However, adding gD to the gB vaccine did not result in significantly better protection. Based on these data, immunogenicity studies in seals under rehabilitation were performed with the gB vaccine only. To this end, gB vaccine was tested at two different doses (20 or 40 microg). PhHV-1 specific antibody titres and in vitro proliferative T cell responses were measured in all seals upon vaccination. No differences were observed in antibody titres between seals vaccinated with either 20 or 40 microgB, but the higher gB concentration did result in higher specific proliferative T cell responses (P<0.01). Based on the close antigenic relationship between PhHV-1 and FHV, the safety and efficacy data in the FHV-cat model, and the immunogenicity data in the vaccinated seals, the gB based vaccine is expected to be safe and effective in protecting against PhHV-1 related disease in harbour seals.

Animals↗
PubMed2003

Protection and antibody responses to oral priming by attenuated human rotavirus followed by oral boosting with 2/6-rotavirus-like particles with immunostimulating complexes in gnotobiotic pigs.

We evaluated antibody responses and protection induced by attenuated Wa human rotavirus (AttHRV) and VP2/6-rotavirus-like particles (VLP), 100 or 250 microg/dose, with immunostimulating complexes (ISCOM) (VLP/ISCOM) each given orally, alone or sequentially to gnotobiotic pigs. The AttHRV-VLP 250 microg/ISCOM and three-dose-AttHRV (AttHRV3x) groups had significantly higher serum IgA, IgG and intestinal IgA antibody titers to HRV pre-challenge than the three-dose-VLP 100 microg/ISCOM group (VLP/ISCOM3x) and controls (diluent/ISCOMmatrix). Protection rates against viral shedding and diarrhea were highest in the AttHRV-VLP250 microg/ISCOM and AttHRV3x groups, lower in the AttHRV-VLP 100 microg/ISCOM group, with no protection in the VLP/ISCOM3x group and controls. Thus, VLP/ISCOM boosted antibody titers and protection after priming with AttHRV.

Adjuvants, Immunologic↗
PubMed1999

Highly immunogenic and fully synthetic peptide-carrier constructs targetting GnRH.

To use peptides as synthetic vaccines, they have to be coupled to a carrier protein to make them more immunogenic. Coupling efficiency between a carrier protein and a peptide, however, is difficult to control with respect to loading density of the peptide. This makes these carrier proteins poorly suitable for practicle use. Attempts have been reported to find carrier molecules or delivery systems which allow easy coupling or incorporation of peptides, reproducible loading densities and well defined products. We compared several promising constructs or delivery systems by immunization of male pigs using a tandem GnRH peptide as a branched polylysine construct, a lipo-thioester, a lipo-amide or a KLH conjugate in CFA, and the lipoamide peptide in an immuno-stimulating complex (ISCOM). We found the lipo-thioester and the branched polylysine constructs to be the most effective carrier molecules for the induction of antibodies against GnRH and immunocastration of pigs.

Adjuvants, Immunologic↗
PubMed2002

Use of bulk milk for detection of Neospora caninum infection in dairy herds in Thailand.

The relationship between the level of Neospora caninum antibodies in bulk milk and the seroprevalence in lactating cows was investigated. Bulk milk was also used to estimate the prevalence of N. caninum infection in dairy herds in the northeast and north Thailand. Bulk milk and individual serum from all lactating cows in 11 herds as well as 220 bulk milk samples from nine milk collection centres were analysed for presence of N. caninum antibodies using an iscom ELISA. In the 11 herds the bulk milk absorbances ranged between 0.04 and 0.89 and the seroprevalences varied between 0 and 46%. Five herds had milk absorbances below 0.20, among those were the two herds housing only seronegative lactating cows. In the remaining three herds with such low bulk milk absorbances one or two cows (5-14%) were seropositive. Six of the investigated herds had bulk milk absorbances above 0.20. In the two herds with the highest bulk milk absorbances more than 30% of the cows were seropositive. Using an absorbance of 0.20 to discriminate between negative and positive herds, 102 (46%) of 220 bulk milk samples were judged positive. There was no significant difference in mean bulk milk absorbance between the milk collection centres within each region. However, the proportion of herds with bulk milk absorbances > or =0.50 in the north was statistically (P < 0.01) higher than that in the northeast. It was concluded that bulk milk antibody testing can be used to identify N. caninum-infected herds and that N. caninum is a common infection in dairy herds in Thailand.

Animals↗
PubMed1999

Serum antibody profile and reproductive performance during two consecutive pregnancies of cows naturally infected with Neospora caninum.

The objective of this study was to record how the antibody levels change over time during pregnancy in dairy cows naturally infected with the protozoan parasite Neospora caninum, and relate this to the reproductive performance. Eighteen cows with antibodies to N. caninum were serum sampled monthly during their first pregnancy and 13 of them were also followed for a second pregnancy. In all, five pregnancies ended in abortion and two in stillbirth. Antibodies to N. caninum in serum were demonstrated by immune stimulating complex enzyme-linked immunosorbent assay (iscom ELISA). The N. caninum antibody titres remained well above the 1:100 cut-off limit for the test used during 2 years in all cows. In the non-aborting cows, mean values of antibody titres to N. caninum rose 1.5-2.5 dilution steps to reach a plateau 4-5 months before parturition, and thereafter decreased from 2 months before parturition. These changes were statistically significant (p < 0.001). The same pattern was seen in the aborting cows. The consistent pattern of rise in antibody titres observed during both pregnancies in all cows indicated a reactivation rather than a reinfection of the parasite at mid-gestation.

Animals↗
PubMed2002

Recent advances in vaccine adjuvants.

New generation vaccines, particularly those based on recombinant proteins and DNA, are likely to be less reactogenic than traditional vaccines but are also less immunogenic. Therefore, there is an urgent need for the development of new and improved vaccine adjuvants. Adjuvants can be broadly separated into two classes based on their principal mechanisms of action: vaccine delivery systems and immunostimulatory adjuvants. Vaccine-delivery systems generally are particulate (e.g., emulsions, microparticles, iscoms, and liposomes) and function mainly to target associated antigens into antigen-resenting cells. In contrast, immunostimulatory adjuvants are derived predominantly from pathogens and often represent pathogen-ssociated molecular patterns (e.g., lipopolysaccaride, monophosphoryl lipid A, CpG DNA). which activate cells of the innate immune system. Recent progress in innate immunity is beginning to yield insight into the initiation of immune responses and the ways in which immunostimulatory adjuvants may enhance this process. The discovery of more potent adjuvants may allow the development of prophylactic and therapeutic vaccines against cancers and chronic infectious diseases. In addition, new adjuvants may also allow vaccines to be delivered mucosally.

Adjuvants, Immunologic↗
PubMed1999

Advances in vaccine adjuvants.

Currently, aluminum salts and MF59 are the only vaccine adjuvants approved for human use. With the development of new-generation vaccines (including recombinant subunit and mucosal vaccines) that are less immunogenic, the search for more potent vaccine adjuvants has intensified. Of the novel compounds recently evaluated in human trials, immunostimulatory molecules such as the lipopolysaccharide derived MPL and the saponin derivative QS21 appear most promising, although doubts have been raised as to their safety in humans. Preclinical work with particulate adjuvants, such as the MF59 microemulsion and lipid-particle immune-stimulating complexes (Iscoms), suggest that these molecules are also potent elicitors of humoral and cellular immune responses. In addition, preclinical data on CpG oligonucleotides appear to be encouraging, particularly with respect to their ability to selectively manipulate immune responses. While all these adjuvants show promise, further work is needed to better define the mechanisms of adjuvant action. Ultimately, the development of more potent adjuvants may allow vaccines to be used as therapeutic, rather than prophylactic, agents.

Adjuvants, Immunologic↗
PubMed1996

Modulation of immune responses by bovine beta-casein.

The present reports the influence of bovine beta-casein on in vitro and in vivo immune responses. Bovine beta-casein showed an inhibitory effect on ovine neutrophil chemotaxis but had an enhancing effect on superoxide production by neutrophils. In response to mitogenic stimulation, the proliferative response of both T and B lymphocytes was significantly enhanced by beta-casein. While beta-casein had no significant effects on IFN gamma production by ovine blood lymphocytes, and TNF alpha production and MCH Class II antigen expression by ovine bronchoalveolar macrophages, it enhanced IL-1 beta production by the macrophages, beta-casein also had no influence on bovine NK cell activity against a virally-infected cell line. Interestingly, beta-casein was found to reduce the adjuvant effect of matrix immune stimulating complexes (ISCOM) on anti-ovalbumin antibody response in mice when given intramuscularly. Taken together, the results suggest that bovine beta-casein had selective modulating effects in vitro on both innate and adaptive immune responses in ruminants, whereas systemic administration of beta-casein, that might have a depressive effect on adjuvant activity, requires further study.

Animals↗
PubMed1996

The flagellar fraction of Trypanosoma cruzi depleted of an immunosuppressive antigen enhances protection to infection and elicits spontaneous T cell responses.

The flagellar fraction (FF) of Trypanosoma cruzi can be separated by immunoaffinity chromatography in two fractions with balanced but opposite immunological effects. The immunoaffinity purified fraction has immunosuppressive activity mediated at least partially by TGF-beta (Hansen et al., submitted). Here we report that the fraction depleted of immunosuppresive antigens (FT) administered with iscom-matrix as adjuvant provides enhanced protection to an infection challenge in immunized mice. In vitro, the FT but not the FF stimulated resident peritoneal cells to produce IL-1 and IL-6. In immunized mice, the FT elicited higher levels of antigen-specific IgG2a than the FF as well as broader recognition of T. cruzi antigens. Splenocytes from mice immunized with FT proliferated spontaneously in vitro and secreted TH1 and TH2 cytokines. The protection provided by FT correlates with its capacity to enhance the secretion of IFN-gamma. We postulate that immunosuppressive antigens present in the FF prevent the development of memory cells secreting IFN-gamma through a TGF-beta dependent mechanism.

Adjuvants, Immunologic↗
PubMed1999

Evaluation of recombinant sporozoite antigen SPAG-1 as a vaccine candidate against Theileria annulata by the use of different delivery systems.

The major sporozoite surface antigen of Theileria annulata (SPAG-1) is a candidate for inclusion in a subunit vaccine. In this paper we summarize the results of 4 vaccination experiments using recombinant SPAG-1 expressed in different systems and presented in different adjuvants. The antigen has been presented as either a C terminal 108 amino acid peptide (called SR1) expressed as both beta-galactosidase and hepatitis B core antigen fusions or as a full-length form expressed as a GST fusion with an N terminal His6 tag. We used different adjuvants, namely Freund's, saponin, ISCOMs and a proprietary adjuvant supplied by SmithKline Beecham, which we call SKBA. The data point to the conclusion that SPAG-1 can elicit partial protection and is therefore suitable for inclusion in an eventual multicomponent subunit vaccine.

Animals↗