Anti-HBV DNA vaccination does not prevent relapse after discontinuation of analogues in the treatment of chronic hepatitis B: a randomised trial--ANRS HB02 VAC-ADN.

OBJECTIVE: The antiviral efficacy of nucleos(t)ide analogues whose main limitation is relapse after discontinuation requires long-term therapy. To overcome the risk of relapse and virological breakthrough during long-term therapy, we performed a phase I/II, open, prospective, multicentre trial using a HBV envelope-expressing DNA vaccine. DESIGN: 70 patients treated effectively with nucleos(t)ide analogues for a median of 3 years (HBV DNA <12 IU/mL for at least 12 months) were randomised into two groups: one received five intramuscular injections of vaccine (weeks 0, 8, 16, 40 and 44) and one did not receive the vaccine. Analogues were stopped after an additional 48 weeks of treatment in patients who maintained HBV DNA <12 IU/mL with no clinical progression and monthly HBV DNA for 6 months. The primary endpoint was defined as viral reactivation at week 72 (HBV DNA >120 IU/mL) or impossibility of stopping treatment at week 48. RESULTS: Reactivation occurred in 97% of each group after a median 28 days without liver failure but with an HBV DNA <2000 IU/mL in 33%; 99% of adverse reactions were mild to moderate. Immune responses were evaluated by enzyme-linked immunosorbent spot and proliferation assays: there was no difference in the percentage of patients with interferon-γ secreting cells and a specific T-cell proliferation to HBcAg but not to HBsAg after reactivation in each group. CONCLUSIONS: Although it is fairly well tolerated, the HBV DNA vaccine does not decrease the risk of relapse in HBV-treated patients or the rate of virological breakthrough, and does not restore the anti-HBV immune response despite effective viral suppression by analogues. TRIAL REGISTRATION NUMBER: NCT00536627.

Immunological and antiviral responses after therapeutic DNA immunization in chronic hepatitis B patients efficiently treated by analogues.

A substudy of a phase I/II, prospective, multicenter clinical trial was carried out to investigate the potential benefit of therapeutic vaccination on hepatitis B e antigen-negative patients with chronic hepatitis B (CHB), treated efficiently with analogues. Patients were randomized in 2 arms, one receiving a hepatitis B virus (HBV) envelope DNA vaccine, and one without vaccination. At baseline, HBV-specific interferon (IFN)-γ-producing T cells were detected in both groups after in vitro expansion of peripheral blood mononuclear cells. Vaccine-specific responses remained stable in the vaccine group, whereas in the control group the percentage of patients with HBV-specific IFN-γ-producing T cells decreased over time. The vaccine-specific cytokine-producing T cells were mostly polyfunctional CD4(+) T cells, and the proportion of triple cytokine-producer T cells was boosted after DNA injections. However, these T-cell responses did not impact on HBV reactivation after stopping analogue treatment. Importantly, before cessation of treatment serum hepatitis B surface antigen (HBsAg) titers were significantly associated with DNA or HBsAg clearance. Therapeutic vaccination in CHB patients with persistent suppression of HBV replication led to the persistence of T-cell responses, but further improvements should be searched for to control infection after treatment discontinuation.

Impact of the Immunogen Nature on the Immune Response against the Major HBV Antigens in an HBsAg and HLA-humanized Transgenic Mouse Model.

Hepatitis B chronic carriage remains as a major public health problem. Protein and DNA vaccines are now widely used in therapeutic vaccine candidates. Although, the hepatitis B surface antigen (HBsAg) based vaccines have been largely studied, candidates comprising both HBsAg and core (HBcAg) either protein- or DNA-based approaches deserve further immunological characterization. In the present study, a repeated dose administration schedule for protein or DNA immunogens was conducted in order to characterize the resulting immune response in a humanized and HBsAg-tolerized setting. A novel transgenic (Tg) mice that express the HBsAg, human MHC class I (HLA-A*0201) and MHC class II (HLA-DRB1*01) molecules and devoid of endogenous murine class I and II molecules was used as a model of HBV chronic carrier. Mice were immunized by subcutaneous (protein) or intramuscular (DNA) routes and the humoral and cellular responses were evaluated. Protein or DNA immunization induced humoral immune responses against both HBsAg and HBcAg. The systematic analysis of epitopes that activate CD4+ and CD8+ T lymphocytes confirmed the accuracy of the model. Cellular immune responses were detected differing in their nature. CD8 T-cell responses were induced mostly after DNA immunization while CD4 T-cell responses were predominant in protein based immunizations. In addition, the intensity of HLA-A2-restricted CD8+ T cell responses was reduced in Tg mice expressing HBsAg when compared to control Tg mice. In conclusion, our results indicate that cellular immune responses necessary for the development of protective immunity can be achieved by DNA or protein immunization. However, important differences in their nature arise when immunogens are administered several times. How to cite this article: Mancini-Bourgine M, Guillen G, Michel ML, Aguilar JC. Impact of the Immunogen Nature on the Immune Response against the Major HBV Antigens in an HBsAg and HLA-humanized Transgenic Mouse Model. Euroasian J Hepato-Gastroenterol 2014;4(1):36-44.

HBsAg/HLA-A2 transgenic mice: a model for T cell tolerance to hepatitis B surface antigen in chronic hepatitis B virus infection.

A humanized murine model was developed to study T cell tolerance to the hepatitis B surface antigen (HBsAg) that is present in sera of hepatitis B virus chronic carriers. The HBsAg/HLA-A2 double-transgenic mice express a chimeric HLA-A2 MHC class I molecule and a high amount of the HBsAg in the liver that is secreted and present in sera during the animal's lifetime. In these mice, injection of plasmid DNA encoding HBsAg induced a high frequency of CD8(+) T cells secreting IFN-gamma in the periphery, with in vitro cytolytic activity and specificity for two dominant HBs-specific HLA-A2-restricted epitopes. Nevertheless, the DNA-based immunization elicited neither T(h)1 nor T(h)2 CD4(+) T cell responses. Despite a high concentration of HBsAg in sera, these mice developed an immunocompetent CD8(+) T cell repertoire towards the viral self-antigen, whereas the CD4(+) T cell repertoire was tolerized. In the absence of a CD4(+) T cell response, the IFN-gamma-secreting CD8(+) T cells primed by DNA-based immunization were unable to exert their antiviral functions in vivo on liver cells expressing the transgene product. However, when pro-inflammatory stimuli were given before or after DNA-based immunization, the HBsAg was cleared from the serum. This effect was antibody dependent and associated with the detection of an HBs-specific T(h)1 CD4(+) T cell response in the periphery. This model provides evidence that HBsAg displayed a strong tolerogenic effect on the CD4(+) T cell compartment that is associated with a defect in CD8(+) T cell effector functions in vivo.

Sex as a variable can be a surrogate for some working conditions: factors associated with sickness absence.

More than twice as many workdays are lost to illness than for personal or family reasons. We examine possible workplace determinants of sickness absence among French workers in the food processing industry. These workers are exposed to a variety of environmental and organizational constraints: cold, uncomfortable postures, assembly-line work, and irregular schedules. In 1987-1988, a medical examination and questionnaire were administered to 558 men and 790 women as part of a study of 17 poultry slaughterhouses and 6 canning factories. Women's and men's working conditions were very different, and their sickness absences for musculoskeletal and respiratory illnesses were related to some of their specific working conditions: cold exposure, ill-adapted work stations, and problems with their supervisors and co-workers. If male and female workers were combined into a single analysis that adjusted for sex, many of the associations operant for a single sex could no longer be seen.

Risk factors for Raynaud's phenomenon among workers in poultry slaughterhouses and canning factories.

BACKGROUND: Apart from the use of vibrating tools, little is known about risk factors for Raynaud's phenomenon. However, it has been hypothesized that this disorder may have a multifactorial aetiology, involving potential causal or triggering factors which can be found in the workplace. The objective of the study is to identify individual and occupational risk factors of Raynaud's phenomenon in a population of workers not exposed to vibration, but exposed to cold. METHODS: The survey was carried out in 1987-1988 in 17 poultry slaughterhouses and six canning factories and included 1474 employees. Data were collected at the annual visit to the occupational health physician. Finger sensitivity to cold and Raynaud's phenomenon were identified from a list of symptoms occurring from exposure to cold. The role of potential risk factors was assessed using multiple logistic regression. RESULTS: A high prevalence of symptoms of finger sensitivity to cold was observed. Raynaud's phenomenon was more common in women than in men, was related to family history of the disease but not to smoking or alcohol consumption. After controlling for non-occupational factors, the following working conditions appeared as risk factors for Raynaud's phenomenon: use of plastic gloves, less than four rest breaks, breaks in an unheated place, continual repetition of the same series of operations, exertion of the arm or hand and being able to think of something else while working. CONCLUSION: The study showed that a number of working conditions were associated with an increased risk of Raynaud's phenomenon and finger sensitivity to cold. Changes in working conditions might reduce the risk of this disorder in the food processing industry.

Conformity of Workers' and Occupational Health Physicians' Descriptions of Working Conditions.

To determine whether workers, describing their own work, and occupational physicians, describing typical workstations, would report the same working conditions, and whether the relationships between health status and working conditions described by these two sources would be similar. An epidemiologic survey was carried out in 1987-88 in 17 poultry slaughterhouses and six canneries in France. The data were collected in two ways: workers described their own working conditions, and occupational physicians described the working conditions at workstations in the same factories. The study included 507 workers who worked at fixed workstations that had been described by the 24 occupational physicians. Health data were obtained from the workers by the physicians during their annual visits. The agreement between workers and physicians in the descriptions of seven working conditions was analyzed. On the whole, the prevalences of exposures to the seven selected working conditions estimated by the two sources were similar. Nevertheless, the observed agreement was not necessarily high. When agreement existed about specific working conditions, the relationships observed between those conditions and workers' health were generally similar for the two sources. This result was stronger for physical health problems than for mental health problems. When both sources agreed that the worker was being exposed to a risk factor, the risk estimation was higher compared with the cases of discordant judgment. This study confirms the usefulness of information provided by experts but also the necessity to interview workers themselves about their working conditions.

Risk factors for high blood pressure among workers in French poultry slaughterhouses and canneries.

The aim of the present study is to examine the relationship between the working conditions of employees in the food industry and blood pressure. An epidemiological survey was conducted between 1987 and 1988 in 17 poultry slaughterhouses and 6 canneries in the French regions of 'Bretagne' and 'Pays de Loire'. One thousand, four hundred and seventy-four workers were included in the study. Data was collected in the course of the medical visit organized annually for employees. Mean diastolic (DBP) and systolic blood pressure (SBP) were analyzed separately for male and female workers. The results indicate a significant relationship between blood pressure, and age and obesity. Amongst the various features of working condition studies, loud noise and the number of work breaks were found to be associated with heightened mean values of DBP or SBP in men only. Type and size of the factory was found to be associated with blood pressure readings for both sexes. A number of working conditions giving rise to heightened mental strain were found to be related to a lowering in mean blood pressure: for example, irregular work finishing times for men and production-line work for women. A discussion of these results reveals the complexity of the relationships which exist between physical and environmental factors in this type of setting and blood pressure of employees.

Factors associated with dysmenorrhea among workers in French poultry slaughterhouses and canneries.

The food and agriculture industry employs 15% of the female industrial work force in France. Workers in this industry are exposed to a variety of environmental and organizational constraints: cold, uncomfortable postures, assembly-line work, irregular schedules. In 1987 to 1988, a medical examination and questionnaire were administered to 726 menstruating women who had not been pregnant during the 2 previous years, as part of a study of French workers in 17 poultry slaughterhouses and 6 canning factories. Dysmenorrhea during the previous year was more prevalent among younger women and smokers, and less prevalent among users of oral contraceptives. After adjustment for nonoccupational variables, dysmenorrhea was significantly related to several parameters expressing cold exposure and physical work load. Other parameters such as job satisfaction and hours of domestic work were not associated with dysmenorrhea.

Menstrual-cycle characteristics and work conditions of workers in poultry slaughterhouses and canneries.

The food and agriculture industry employs 14.6% of the female industrial work force in France. Workers are exposed to a variety of environmental and organizational constraints (eg, irregular schedules, cold, uncomfortable postures, repetitive movements). In 1987-1988 a medical examination and questionnaire were administered to 726 workers with menstrual periods in 17 poultry slaughterhouses and six canning factories. Anomalies (irregular cycles, amenorrhea, long cycles) during the previous year were associated with work conditions. After adjustment for relevant nonoccupational variables, irregular cycles were significantly related to schedule variability and cold exposure, amenorrhea was associated with cold exposure, and long cycles with schedule variability. Other parameters such as repetitive work, standing posture, lifting weights, job satisfaction, and hours of domestic work were not associated with cycle anomalies. Cycle anomalies may be a useful indicator of occupational effects on female reproduction, analogous to the use of sperm parameters to warn of effects on male workers.