Production of Human Parvovirus 4 VP2 Virus-Like Particles in Yeast and Their Evaluation as an Antigen for Detection of Virus-Specific Antibodies in Human Serum

Abstract

Background: Human parvovirus 4 (PARV4) is a recently discovered member of the Parvoviridae family, which is not closely related to any previously discovered human parvoviruses. PARV4 has been isolated from the plasma of individuals with symptoms of acute viral infection; however, until recently PARV4 had not been associated with any disease, and its prevalence in the human population is yet to be established. Methods: The major capsid protein VP2 of PARV4 was generated in the yeast Saccharomyces cerevisiae and used for serological detection of virus-specific IgG and IgM in the sera of low-risk individuals. Results: One hundred and seventy serum specimens obtained from patients with acute respiratory diseases were tested for PARV4-specific IgG and IgM antibodies. Sixteen individuals (9.4%) were diagnosed as seropositive, including 6 IgM and IgG positive, 6 IgM positive/IgG negative and 4 IgG positive/IgM negative. Seven of the 16 seropositive individuals were between the ages of 3 and 11 with no evidence of parenteral exposure to PARV4 infection. Conclusion: Our data demonstrate that recombinant yeast-derived VP2 protein, self-assembled to virus-like particles, can represent a useful tool when studying the seroprevalence of PARV4 infection. The presence of PARV4-specific antibodies in a low-risk group may indicate the possibility of alternative routes of virus transmission.

Viruses of the Parvoviridae Family: Molecular Genetical Aspects of Reproduction and Medical Importance

“…При обследовании больных с ОРЗ в 9,4% случаев были обнаружены IgG-и IgM-АТ к PARV4 [74]. ДНК PARV4 была выделена в спинномозговой жидкости детей первых трех лет жизни с энцефа-литом [10].…”

“…Для создания тест-систем, с помощью вектора, встроенного в клетки дрожжей, получены ви-рус-подобные частицы на основе VP2 белка [74]. Т-клеточный ответ на PARV4 можно опреде-лить in vitro в Interferon-gamma ELISpot иссле-довании.…”

Generation of recombinant porcine parvovirus virus-like particles in Saccharomyces cerevisiae and development of virus-specific monoclonal antibodies

“…To our knowledge, our study provides the first evidence of stable recombinant PPV VP2 VLPs not produced in baculovirus expression system. S. cerevisiae expression system has been shown to be efficient in producing antigenic VLPs of diagnostic relevance of human parvoviruses [28,29] and porcine circovirus [39]. Therefore, it represents a potential system to meet the need for VLP-forming antigens to detect and differentiate a number of newly discovered porcine.…”

“…VLPs generated in baculovirus system exhibit positive immunoreactivity for PPV and are used in most commercial ELISA tests [26]. Most recently, immunogenic PPV VP2 protein was synthesized in yeast Pichia pastoris [27] The formation of recombinant antigenic human parvovirus capsid protein VLPs in S. cerevisiae has been recently demonstrated [28,29]. Regarding costs, yield, and ease of handling, VLP production in yeast represents an alternative to the recombinant baculovirus expression system, which is so far the dominating source of VP2-derived VLPs of parvoviruses [27,28].…”

Generation of Recombinant Porcine Parvovirus Virus-Like Particles inSaccharomyces cerevisiaeand Development of Virus-Specific Monoclonal Antibodies

“…The formation of recombinant antigenic human parvovirus capsid protein VLPs in S. cerevisiae has been recently demonstrated [28, 29]. Regarding costs, yield, and ease of handling, VLP production in yeast represents an alternative to the recombinant baculovirus expression system, which is so far the dominating source of VP2-derived VLPs of parvoviruses [27, 28].…”