Difluoromethylornithine (DFMO), an Inhibitor of Polyamine Biosynthesis, and Antioxidant N-Acetylcysteine Potentiate Immune Response in Mice to the Recombinant Hepatitis C Virus NS5B Protein.

Hepatitis C virus (HCV) is one of the main triggers of chronic liver disease. Despite tremendous progress in the HCV field, there is still no vaccine against this virus. Potential vaccines can be based on its recombinant proteins. To increase the humoral and, especially, cellular immune response to them, more effective adjuvants are needed. Here, we evaluated a panel of compounds as potential adjuvants using the HCV NS5B protein as an immunogen. These compounds included inhibitors of polyamine biosynthesis and urea cycle, the mTOR pathway, antioxidants, and cellular receptors. A pronounced stimulation of cell proliferation and interferon-γ (IFN-γ) secretion in response to concanavalin A was shown for antioxidant N-acetylcysteine (NAC), polyamine biosynthesis inhibitor 2-difluoromethylornithine (DFMO), and TLR9 agonist CpG ODN 1826 (CpG). Their usage during the immunization of mice with the recombinant NS5B protein significantly increased antibody titers, enhanced lymphocyte proliferation and IFN-γ production. NAC and CpG decreased relative Treg numbers; CpG increased the number of myeloid-derived suppressor cells (MDSCs), whereas neither NAC nor DFMO affected MDSC counts. NAC and DFMO suppressed NO and interleukin 10 (IL-10) production by splenocytes, while DFMO increased the levels of IL-12. This is the first evidence of immunomodulatory activity of NAC and DFMO during prophylactic immunization against infectious diseases.

Immune function of zhenqi fuzheng capsule on volleyball players after high intensity training / Função imunitária da cápsula zhenqi fuzheng em jogadores de voleibol após treinamento de alta intensidade / Efecto de la cápsula zhenqi fuzheng sobre la función inmune de los jugadores de voleibol después del entrenamiento de alta intensidad

ABSTRACT At present, many studies have proved that proper exercise can promote the immune function of human body to a certain extent, but athletes need a lot of high-intensity sports training, and their immune function declines instead of improving. In order to control the decline of immune function of athletes after high-intensity training, this study propose the Zhenqi Fuzheng capsule to achieve this goal. Through experimental comparison, the parameters such as white blood cell content, immunoglobulin number, T lymphocyte, human hemoglobin content and exercise exhaustion time were detected after high-intensity training. The results showed that compared with the control group taking Zhenqi Fuzheng, the weight of those who had taken qifuzhengs capsule did not change, and the content of white blood cells, immunoglobulin, hemoglobin content and exercise time increased to a certain extent. The results showed that Zhenqi Fuzheng could inhibit the decrease of body immune function after high-intensity exercise, then accelerate the recovery of human immune function. This study is expected to enhance the immunity of sports athletes, and reduce athletes' pain after high-intensity training.

RESUMO Atualmente, muitos estudos prova que exercícios adequados podem promover a função imunológica do corpo humano em certa medida, mas os atletas precisam de muito treinamento esportivo de alta intensidade, e sua função imunológica diminui em vez de melhorar. A fim de controlar o declínio da função imunológica dos atletas após treinamento de alta intensidade, este estudo propôs a administração da cápsula Zhenqi Fuzheng para alcançar esse objetivo. Através de comparação experimental, foram detectados parâmetros como o teor de glóbulos brancos, imunoglobulina, linfócitos T, hemoglobina humana e tempo de exaustão do exercício após treinamento de alta intensidade. Os resultados mostraram que, em comparação com o grupo controle que tomou a cápsula Zhenqi Fuzheng, o peso daqueles que tinham tomado a cápsula de qifuzheng não se alterou, e o teor de glóbulos brancos, imunoglobulina, hemoglobina e o tempo de exercício aumentaram em certa medida. Os resultados mostraram que a cápsula Zhenqi Fuzheng poderia inibir a diminuição da função imunológica corporal após exercícios de alta intensidade, e acelerar a recuperação da função imunológica humana. Espera-se que este estudo possa aumentar a imunidade dos atletas e reduzir a dor dos atletas após treinamento alta intensidade para fornecer uma certa referência.

RESUMEN Actualmente, muchos estudios prueban que ejercicios adecuados pueden promover la función inmunológica del cuerpo humano en cierta medida, pero los atletas precisan mucho entrenamiento deportivo de alta intensidad, y su función inmunológica disminuye en vez de mejorar. A fin de controlar la declinación de la función inmunológica de los atletas después del entrenamiento de alta intensidad, este estudio propuso la administración de la cápsula Zhenqi Fuzheng para alcanzar ese objetivo. Por medio de comparación experimental, fueron detectados parámetros como el tenor de glóbulos blancos, inmunoglobulina, linfocitos T, hemoglobina humana y tiempo de agotamiento del ejercicio después de entrenamiento de alta intensidad. Los resultados mostraron que, en comparación con el grupo control que tomó la cápsula Zhenqi Fuzheng, el peso de aquellos que habían tomado la cápsula de qifuzheng no se alteró, y el tenor de glóbulos blancos, inmunoglobulina, hemoglobina y el tiempo de ejercicio aumentaron en cierta medida. Los resultados mostraron que la cápsula Zhenqi Fuzheng podría inhibir la disminución de la función inmunológica corporal después de ejercicios de alta intensidad, y acelerar la recuperación de la función inmunológica humana. Se espera que este estudio pueda aumentar la inmunidad de los atletas y reducir el dolor después de entrenamiento de alta intensidad para proveer una cierta referencia.

Combining vitamin A and vaccines: convenience or conflict?

The present thesis is based on 11 papers from 1995-2010. The studies have mainly taken place at the Bandim Health Project in Guinea-Bissau, West Africa, but a reanalysis of a randomised trial from Ghana is also included. My research has explored the consequences of combining high-dose vitamin A supplementation and childhood vaccines. Vitamin A deficiency is associated with increased mortality. To protect against the consequences of vitamin A deficiency the World Health Organization recommends that high-dose vitamin A supplements be given together with routine vaccines to children between 6 months and 5 years of age in more than 100 low-income countries. The recommendation is based on logistical considerations. The consequences of combining vitamin A and vaccines were not investigated in randomised trials prior to the implementation of this policy - it was assumed that the interventions were independent. My first project aimed to study the effect on the immune response to measles of providing vitamin A together with measles vaccine. We found that the two interventions were not independent. Vitamin A enhanced the antibody response to measles vaccine given at 9 months of age significantly, especially in boys. The effects were sustained over time; the children who had received vitamin A with their measles vaccine were more protected against measles at 6-8 years of age. Though vitamin A supplementation had a beneficial effect on the immune response to measles vaccine, it intrigued me that the effect of vitamin A supplementation on overall mortality was not always beneficial. While vitamin A was beneficial when given after 6 months of age, and two studies had shown a beneficial effect when given at birth, all studies testing the effect between 1-5 months of age had found no effect. These time windows are dominated by three different childhood vaccines: BCG vaccine given at birth, diphtheria-tetanus-pertussis (DTP) vaccine given between 1-5 months of age, and measles vaccine given at 9 months of age. These vaccines have been shown to have strong effects on mortality from infectious diseases in general, so-called non-specific effects. The live BCG and measles vaccine protects against more mortality than can be ascribed to the prevention of tuberculosis and measles, respectively. The inactivated DTP vaccine worryingly has been associated with increased mortality from other infectious diseases. Both positive and negative effects are strongest for girls. I proposed the hypothesis that vitamin A amplifies not only the specific vaccine effects, as we saw for measles vaccine, but also the non-specific effects of vaccines on mortality from other infectious diseases. According to my hypothesis, vitamin A would enhance the non-specific beneficial effects on mortality of BCG and measles vaccine, but also the negative effects of DTP vaccine. Hence, the hypothesis offered an explanation for the mortality-age pattern after vitamin A supplementation. Since it was formulated, I have aimed to test this hypothesis. Since it is associated with ethical problems to randomise children above 6 months of age to vitamin A supplementation, and to randomise children in general to recommended vaccines, we have had to be pragmatic when designing the trials. Hence, our studies have taken many different forms. We conducted an observational study during a vitamin A campaign in which missing vaccines were also provided, and a randomised trial testing the effect of two different doses of vitamin A during another campaign; we tested the effect of providing vitamin A with BCG at birth in two randomised trials, and we reanalysed data from one of the original randomised trials of vitamin A supplementation from the perspective of vaccination status. In all studies the main outcome was mortality. The results document that vitamin A supplements do more than protect against vitamin A deficiency. They support the hypothesis that vitamin A supplements interact with vaccines with important consequences for mortality. First, a smaller dose of vitamin A was more beneficial than a larger dose for girls. Second, the effect of vitamin A given with DTP vaccine was significantly different from the effect of vitamin A given with measles vaccine, and children, who received vitamin A with DTP vaccine, had higher mortality than children, who had received vitamin A alone, or who did not receive anything. Third, vitamin A given with BCG at birth interacted negatively with subsequent DTP vaccines in girls. Fourth, the effect of vitamin A to older children in Ghana depended on vaccination status, being beneficial in boys, but harmful in girls who received DTP vaccine during follow-up. The results also show that boys and girls respond differently to vitamin A and vaccines. It is a common assumption within public health in low-income countries that interventions can be combined without producing unexpected consequences. The work presented in this thesis confronts this assumption; the results show that vitamin A and vaccines should be seen not only as specific interventions with specific and independent effects, but as immuno-modulators, which can interact with important consequences for overall mortality. Combining interventions can be convenient and lead to synergistic health benefits, but we documented several examples, where it also leads to unexpectedly increased mortality. Thus, to optimise the child health intervention policy in low-income countries a shift in paradigm is needed. Health interventions should no longer be seen as merely specific and independent, and the policy should probably not be the same for boys and girls. Though more complex, it is necessary to evaluate all health interventions in terms of their effect on overall mortality - and their potential interactions with other health interventions and potential sex-differential effects should always be investigated. Only in this way can we assure that the children in the poorest countries get the best possible treatment and avoid using large amounts of money and resources on interventions which may, in worst case, kill them.

Effect of parenteral vitamin E and selenium supplementation on immune status of dogs vaccinated with subunit and somatic antigens against Taenia hydatigena.

Thirty clinically healthy dogs were divided into five equal groups; Gs 1 and 2 were vaccinated with subunit and somatic antigens (Ags) respectively in combination with vitamin E and selenium (Vit E/Se) supplement, Gs 3 and 4 were vaccinated with subunit and somatic Ags respectively and group 5 was kept unvaccinated as control positive. Dogs in the vaccinated-Vit E/Se supplemented groups had a significantly greater (P < 0.05) serum Se and alpha-tocopherol than un-supplemented Gs. Best immune response against T. hydatigena was observed in the Vit E/Se supplemented groups, as evidenced by increase production of antibody titer and IgG concentration in comparison with either vaccinated un-supplemented or control groups. Moreover, the highest protection level against T. hydatigena infection was observed in groups 1 and 2 (83.3%), while was 66.7% and 50% in groups 3 and 4 respectively. Subunit Ag was more efficient than somatic Ag in improving the immune status of dogs. Vit E and Se proved to be immuno-potentiating to dogs vaccinated with subunit and somatic Ags and increased the possibility for the protection against T. hydatigena infection.

Differential effect of phosphodiesterase IV inhibitor RP73401 on various inflammatory and immune responses relevent to rheumatoid arthritis.

Phosphodiesterase (PDE) IV inhibitors have been reported to possess potent anti-inflammatory activities through enhancement of cAMP. In this study, the immunopharmacological effect of PDE IV inhibitor (RP73401) was further carefully evaluated. RP73401 strongly blocked the production of tumor necrosis factor (TNF)-alpha from lipopolysaccharide (LPS)-stimulated murine macrophages (RAW264.7) and human peripheral blood mononuclear cells (PBMC) and LPS-primed mice. RP73401 did not relieve joint inflammation in adjuvant-arthritis (RA) model, whereas the compound attenuated arachidonic acid-induced inflammation. RP73401 displayed weak or no modulatory effects on the activation of macrophage and lymphocytes (assessed by proliferation, nitric oxide (NO) release and cell-cell adhesion, TNF-alpha production upon phorbol 12-myristate 13-acetate (PMA) treatment), and fluorescein-isothiocynate (FITC)-induced ear oedema. Collectively, these data suggest that PDE IV inhibitor RP73401 may differentially modulate various immune responses and these may explain its inability to inhibit adjuvant-induced joint inflammation or FITC-induced ear oedema.

Androstenediol-induced restoration of responsiveness to influenza vaccination in mice.

Androstenediol (AED), a metabolite of dehydroepiandrosterone (DHEA) regulates innate and adaptive immune responses. To examine whether AED could effectively reverse the age-associated decline of antiviral immunity, 3-, 10-, and 22-month-old mice were treated with AED-sulfate (AED-S) for 45 days beginning 10 days prior to vaccination. Subsequently, mice were primed and boosted with suboptimal doses of a commercially-available trivalent influenza vaccine. Treatment of 10-month-old animals with AED-S during vaccination increased the titer of circulating antiviral immunoglobulin G to levels comparable with those in 3-month-old mice. Furthermore, AED-S treatment protected 10-month-old animals from intranasal challenge with a lethal dose of influenza virus 21 days after secondary vaccination. Although AED-S treatment of 22-month-old mice did not enhance vaccine responses and failed to protect against lethal challenge, the data from the 10-month-old animals suggest that treatment with AED-S will prevent the early manifestations of immunosenescence.

Enhancement of respiratory immunity to Pasteurella multocida by cholera toxin in rabbits.

Cholera toxin (CT) is a potent adjuvant for the mucosal immune system. The purpose of this study was to determine if coadministration of CT with a potassium thiocyanate extract of Pasteurella multocida (PTE) leads to enhanced anti-PTE antibody activity and increased protection of rabbits against infection with P. multocida and associated disease. Groups of rabbits were immunized intranasally on days 0, 7, and 14, with phosphate buffered saline (PBS), 200 micrograms of CT, 1.0 mg of PTE, or 1.0 mg PTE with 200 micrograms CT. Nasal lavage and serum samples were collected over 28 days after initial immunization and evaluated by ELISA for specific antibody directed against PTE. Marked increases in serum (IgG) and nasal lavage (IgA) anti-PTE antibody activity were found beginning after day 14 in rabbits immunized with PTE. Rabbits immunized with PTE and CT demonstrated further increases in this activity. Tracheobronchial lavage samples collected at the time of necropsy demonstrated a significant level of anti-PTE IgA activity in animals immunized with PTE, and coadministration with CT stimulated a further significant increase in this activity. Groups of similarly immunized rabbits were challenged 16 days after initial immunization with 5 x 10(7) CFUs of P. multocida. Nasal lavage samples were cultured for P. multocida over the next 10 days. Rabbits were euthanized within 10 days after challenge, tissues cultured for P. multocida, and histopathologic lesion severity graded using a numeric scale. Rabbits immunized with PTE survived longer, had less severe lesions of the lungs, pleura, and liver, and fewer P. multocida CFUs cultured from samples than PBS or CT controls. Coadministration of CT led to further reductions in lesion severity of those tissues and numbers of P. multocida CFUs cultured from samples. Increased nasal turbinate atrophy of rabbits immunized with PTE with or without CT was associated with increased mean survival time. In summary, coadministration of CT with PTE enhanced protective immunity to P. multocida disease and infection in rabbits.

[The development of a new generation of pertussis vaccine]. / Razrabotka kokliushnoi vaktsiny novogo pokoleniia.

The results of the weight gain test on mice have shown that acellular pertussis vaccine is less toxic than the pertussis component of adsorbed diphtheria-pertussis-tetanus (DPT) vaccine due to a lower content of endotoxin in the acellular vaccine; but the leukocytosis-promoting and histamine-sensitizing activities of JNIH-6 and adsorbed DPT vaccines are indicative of incomplete inactivation of Bordetella pertussis toxin. The content of incompletely inactivated B. pertussis toxin is practically the same in both preparations, constituting 1/100-1/200 of the calculated initial activity. For this reason, the use of the new pertussis vaccine also involves a risk of development of serious postvaccinal reactions and/or complications caused by this toxin. Search for the optimum method of inactivation of B. pertussis main toxin should be continued. As shown by the enzyme immunoassay, acellular pertussis vaccine used in the same immunizing dose as adsorbed DPT vaccine induces a more intensive immune response to hemagglutinin and B. pertussis toxin. This is due to higher residual toxicity of the corpuscular component of adsorbed DPT vaccine. Induction of antibodies to B. pertussis toxin has been shown to decrease in response to injection of acellular pertussis vaccine containing a certain residual amount of incompletely inactivated B. pertussis toxin.

Effects of parenteral selenium and vitamin E on performance, health and humoral immune response of steers new to the feedlot environment.

Five trials with steers new to the feedlot environment were conducted to determine the effects of one or two i.m. injections of selenium (Se) and(or) vitamin E (Vit E) on performance, health status and serum antibody response to Pasteurella haemolytica vaccination. In all trials, performance and average number of days sick per steer were not affected (P greater than .05) by single injection of Se and(or) Vit E. In Trial 1, 26 steers (avg initial wt 267 kg) were treated with 1) no Se or Vit E or 2) 25 mg Se (as Na2SeO3) plus 340 IU Vit E (as [d]-alpha-tocopheryl acetate). P. haemolytica serum immunoglobulin G (IgG) titers on d 7 and 14 were greater (P less than .05) for steers receiving 25 mg Se plus 340 IU Vit E. In Trial 2, 141 steers (avg initial wt 204 kg) were treated with 1) no Se or Vit E, 2) 25 mg Se, 3) 340 IU Vit E or 4) 25 mg Se plus 340 IU Vit E. Serum IgG titers were greater (P less than .05) only for Treatment 4 on d 6. Trial 3 was conducted using 107 steers and the same treatments as in Trial 2. By d 14, titers for treatment 4 were greater (P less than .05) than those for Treatments 1 or 3, but not greater than those for Treatment 2. In Trial 4, serum IgG titers were unaffected (P greater than .05) when 48 steers (avg initial wt 248 kg) were treated with 1) no Se or Vit E, 2) 25 mg Se plus 340 IU Vit E 14 d prior to shipping or 3) 25 mg Se plus 340 IU Vit E 14 d prior to shipping, plus repeat injection on day of arrival at the feedlot. In Trial 5, 107 steers were treated with 1) no Se or Vit E, 2) 25 mg Se plus 340 IU Vit E or 3) 50 mg Se plus 680 IU Vit E. Serum IgG titers increased linearly (P less than .01) due to treatment on d 7, 13 and 20 and a quadratic response (P less than .05) was observed on d 27. In these trials, serum antibody response to P. haemolytica vaccination was enhanced with the combination of Se and Vit E; however, performance and health status were not affected.

Protective mechanism of the immune response to a ribosomal vaccine from Pseudomonas aeruginosa. I. In vivo protection studies in compromised animal models.

Studies of the protective activity of a ribosomal vaccine from Pseudomonas aeruginosa in various immunocompromised animal models were performed. The results obtained demonstrated that the vaccine was highly effective in complement (C5)-deficient mice, C3-deficient (cobra venom factor-treated) mice, and leukopenic mice in providing protection against lethal infection with P. aeruginosa. Passive immunization with specific antiserum to the ribosomal vaccine was also effective in leukopenic mice. In mice that were both C5-deficient and leukopenic, the vaccine did not protect mice against lethal infection, but did prolong their survival, whereas in C3-deficient, leukopenic mice significantly enhanced protection was observed. These results were interpreted to suggest the involvement of multiple factors in the protective immune response to the vaccine, including the bactericidal and opsonic activities of specific antibody plus complement and the phagocytic activity of polymorphonuclear leukocytes with opsonized bacteria. Compensation for deficiencies in some of these factors can be obtained by enhancement of other factors through active or passive immunization.

[Experimental immunological effectiveness and safety of pyoimmunogen vaccine against Pseudomonas aeruginosa infection]. / Immunologicheskaia éffektivnost' i bezvrednost' v éksperimente pioimmunogena-vaktsiny protiv infektsii Pseudomonas aeruginosa.

Pyoimmunogen, a polycomponent vaccine against P. aeruginosa infection, has been obtained in laboratory and semi-industrial conditions. The microbial biomass obtained from the strains belonging to O-serotypes (immunotypes) most frequently occurring in clinical practice has been used for producing protective antigens. The preparations have been found to contain proteins (peptides) and carbohydrates in the ratio 6 : 1 to 8 : 1, as well as traces of 2-keto-3-desoxyoctanate, which is indicative of the low content of endotoxin. The immunogenicity of the preparations has been studied experimentally by the active immunization of mice. In these experiments the animals vaccinated in a single injection were found to be protected from challenge with both homologous and heterologous P. aeruginosa strains. The high level of protection from infection caused by toxigenic strain PA-103 was registered. The preparations have low toxicity: LD50 for mice exceeds 2 mg (in protein content): after the multiple administration (7-10 times) of the preparation to mice and rats the weight of the experimental animals was not significantly different from the weight of the control animals.

[Evaluation of the immunogenicity of meningococcal vaccines in experiments on mice]. / Otsenka immunogennosti meningokokkovykh vaktsin v opytakh na myshakh.

The immunogenicity of 2 meningococcal vaccines, multicomponent vaccine produced at the Mechnikov Research Institute for Vaccines and Sera in Moscow and polysaccharide vaccine obtained from Merck Sharp & Dohme (USA), was evaluated on experimental meningococcal sepsis in mice, produced by the injection of meningococcal culture in mucin suspension. The protective effect of these 2 vaccines, expressed in terms of ED50, was 0.28 +/- 0.12 for the multicomponent vaccine and 0.25 +/- 0.24 for the polysaccharide vaccine; the challenge dose used in the test was 10 LD50 of the culture. The multicomponent vaccine gave the maximum immunological effect in a dose of 8 micrograms, while higher or lower doses induced a lesser increase in antibody titer and thus gave lower protection to mice against infection.

[Principle of constructing polyvalent Pseudomonas aeruginosa vaccines]. / Printsip konstruirovaniia polivalentnykh vaktsin Pseudomonas aeruginosa.

Dependence of the range of protective action of P. aeruginosa vaccine on the number of its composites was studied. A principle of the selection of strains who vaccines differed in vivo by immunological specificity was applied to construction of the experimental preparations and modelling a polyvalent vaccine. Increase of the number of components in the vaccine was accompanied by increase of its protective action range. However, with the increase of the number of polyvaccine components in the polyvaccine the accretion of the protective effect expressed in the mean protective index per component displayed a gradual reduction. It was calculated theoretically that a 6--7-component vaccine should provide protection from 94--96% of the P. aeruginosa strains; as to further increase of the number of components--it would induce overloading of the vaccine with a possible absence of any effect.

[Effect of a polyvitamin preparation on the immune response to antibodies. Demonstration of the immunostimulating activity of propylene glycol]. / Activité d'une préparation polyvitaminique sur la réponse immunitaire à anticorps. Mise en évidence de l'activité immuno-stimulante du propylène-glycol

Humoral antibody to sheep erythrocytes was studied by plaque forming cell and hemagglutinin assays in mice receiving a polyvitaminic preparation containing vitamins A, D, E, and C. A statistically significant increase in PFC on days 6, 12 and 18 after immunization, and an increase in hemagglutinin titers was observed in animals treated with this vitamin containing solution. These results are interpreted as an immunostimulating activity of the preparation. Nevertheless, as shown by the results obtained with each component injected separately, a major proportion of the immunostimulating activity seems to be related to propylene glycol, a simple compound contained as an hydrodispersant in the preparation. Further studies of the immunostimulating properties of propylene glycol are now in progress in our laboratory.