Construction and identification of HSV-1 vector vaccine carrying HIV-1 antigen / 生物工程学报

To construct an HSV-1 vector vaccine carrying HIV-1 antigens, HIV-1 gp160, gag, protease and the expression elements were chained together, and then inserted into the internal inverted repeat sequence region of HSV-1 by bacterial artificial chromosome technology. Firstly, HIV-1 gp160 (including type B and C), gag and protease genes were cloned into pcDNA3 in series to generate the pcDNA/gBgp and pcDNA/gCgp, then the recombinant plasmids were transfected into 293FT cells, and HIV-1 antigen was detected from transfected cells by Western blotting. Then the expression cassettes from pcDNA/gBgp and pcDNA/gCgp, comprising HIV-1 antigen genes and expression elements, were cloned into pKO5/BN to generate the shuttle plasmids pKO5/BN/gBgp and pKO5/BN/gCgp. The shuttle plasmids were electroporated into E. coli cells that harbor an HSV-BAC, the recombinant bacteria were screened, and the recombinant DNA was extracted and transfected into Vero cells. The recombinant virus was purified through picking plaques, the virus' DNAs were identified by Southern blotting; HIV-1 antigen was detected from the recombinant HSV-1 infected cells by Western blotting, and the virus' replication competent was analyzed. As the results, gp160 and gag proteins were detected from 293FT cells transfected with pcDNA/gBgp and pcDNA/gCgp by Western blotting. The recombinant bacteria were generated from the E. coli electroporated with pKO5/BN/gBgp or pKO5/BN/gCgp. The recombinant HSV was purified from the Vero cells transfected with the recombinant DNA, the unique DNA fragment was detected from the genome of recombination HSV by Southern blotting; gp120 and gp41 were detected from the infected cells by Western blotting, and the recombinant HSV retained replication competent in mammalian cells. The results indicate that the recombinant HSV carrying HIV-1 gp160, gag and protease genes was generated, the virus retains replication competent in mammalian cells, and could be used as a replicated viral vector vaccine.

Herpes genital / Genital herpes

O herpes genital é uma doença infectocontagiosa sujeita a recidivas, tendo como agente etiológico duas cepas diferentes do vírus herpes simples (HSV), o tipo 1 (HSV-1) e o tipo 2 (HSV-2). A grande maioria dos casos de herpes genital é causada pelo HSV-2, embora a prevalência do HSV-1 esteja em ascensão, principalmente na população jovem e devido à prática de sexo oral. A manifestação clínica pode ser primária ou recorrente, esta última acontecendo por reativação viral. O diagnóstico é feito pelas características clínicas associadas às confirmações laboratoriais da infecção. Diversos estudos clínicos e epidemiológicos demonstram a sinergia entre herpes genital e aids. Na gravidez, a grande preocupação acerca da infecção pelo HSV refere-se à morbidadee à mortalidade associadas à infecção neonatal. Atualmente não existe nenhum tratamento eficaz na cura do herpes genital, mas alguns medicamentos antivirais são capazes de diminuir o tempo da doença e prevenir as erupções. A maioria dos esforços para combater a infecção herpética genital concentrase no desenvolvimento de vacinas.

Genital herpes is an infectious disease subject to recurrent crises, with the etiologic agent of two different strains of herpes simplex virus (HSV), the type 1 (HSV-1) and type 2 (HSV-2). The vast majority of genital herpes cases is caused by HSV-2, although the prevalence of HSV-1 is on the rise, especially in young population and due to oral sex. The clinical manifestations may be primary or recurrent, the latter going on viral reactivation. The diagnosis is made by the clinical characteristics associated with laboratory confirmation of infection. Several epidemiological and clinical studies demonstrate the synergybetween genital herpes and aids. In pregnancy, the major concern about HSV infection refers to the morbidity and mortality associated with neonatal infection. Currently, there is no treatment capable of curing genital herpes, but some antiviral drugs are able to decrease the duration of the disease and prevent flares. Most efforts to combat genital herpes infection are focused on vaccine development.

Status quo of the researches on HSV vaccines / 中华男科学杂志

Herpes simplex virus (HSV) infection can cause severe recurrent disease in humans and establish lifelong latency in the host. Furthermore, it can significantly increase the risk of HIV infection and bring substantial psychosexual as well as physical morbidity to the patients. Several antiviral therapies are available for the control of its symptoms and spreading, but they can neither cure nor alter the nature of HSV infection. The development of an efficacious HSV vaccine is therefore necessitated for controlling the occurrence, transmission and recurrence of the infection. Those that have undergone clinical evaluation include subunit vaccines, attenuated live virus vaccines, replication defective virus vaccine, naked DNA vaccines and viral vector vaccines. Most of the above vaccine strategies have elicited protective immunity in animal models, but none has yet been effective in human beings. For all that, scholars have never stopped their exploration for an effective vaccine against the notorious virus. The authors present an overview of a few most promising candidate vaccines for the prevention and treatment of HSV infection.

Modulation of protective immunity against herpes simplex virus via mucosal genetic co-transfer of DNA vaccine with beta2-adrenergic agonist

Cholera toxin, which has been frequently used as mucosal adjuvant, leads to an irreversible activation of adenylyl cyclase, thereby accumulating cAMP in target cells. Here, it was assumed that beta2-adrenergic agonist salbutamol may have modulatory functions of immunity induced by DNA vaccine, since beta2-adrenergic agonists induce a temporary cAMP accumulation. To test this assumption, the present study evaluated the modulatory functions of salbutamol co-administered with DNA vaccine expressing gB of herpes simplex virus (HSV) via intranasal (i.n.) route. We found that the i.n. co-administration of salbutamol enhanced gB-specific IgG and IgA responses in both systemic and mucosal tissues, but optimal dosages of co-administered salbutamol were required to induce maximal immune responses. Moreover, the mucosal co-delivery of salbutamol with HSV DNA vaccine induced Th2-biased immunity against HSV antigen, as evidenced by IgG isotypes and Th1/Th2-type cytokine production. The enhanced immune responses caused by co-administration of salbutamol provided effective and rapid responses to HSV mucosal challenge, thereby conferring prolonged survival and reduced inflammation against viral infection. Therefore, these results suggest that salbutamol may be an attractive adjuvant for mucosal genetic transfer of DNA vaccine.

Co-expression of HSV-1 gD gene and interleukin-2 gene in dendritic cell / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To construct eukaryotic expression plasmid IRES-gD-IL-2 which contains both HSV-1 glycoprotein D (gD) gene and IL-2 gene and to induce it to express in antigen presenting cell (APC) -- dendritic cells (DCs).</p><p><b>METHODS</b>The whole sequence of gD and IL-2 were amplified by PCR assay. After confirmation by PCR, double-enzyme digestion and sequencing, these genes were directly cloned into eukaryotic expression vector IRES, then were transfected into DCs. Western blotting was employed to identify the transcription and expression of gD gene.</p><p><b>RESULTS</b>The results of PCR and enzyme digestion showed that the recombinant expression plasmid contained correct fragments, and the transcription and expression of gD were confirmed by Western blotting.</p><p><b>CONCLUSION</b>The recombinant expression vector IRES-gD-IL-2 was constructed, the results of the Western blotting showed that the recombinant protein could be identified by gD- specific antibody, therefore the protein has immunologic competence.</p>

Herpes simples - Aspectos clínicos, métodos de diagnósticos e tratamento / Herpes simplex - Clinical features, diagnostic methods and treatment

As lesões causadas pelo vírus herpes simples (HSV) têm prevalência alta entre indivíduos normorreativos e uma prevalência ainda maior nos imunocomprometidos. Os aspectos clínicos da infecção herpética são muito variáveis, principalmente nos pacientes imunodeprimidos, e o herpes freqüentemente é confundido com outras patologias. Por isso, é importante estar atento ao diagnóstico diferencial. Devido ao herpes simples geralmente apresentar uma infecção primária assintomática e, portanto, a maioria das pessoas desconhecerem que estão infectadas, é importante salientar a possibilidade de disseminação assintomática do vírus, que acaba sendo a forma mais importante de transmissão. Com a disponibilidade de testes diagnósticos rápidos e de fácil realização e o constante desenvolvimento de novos agentes antivirais, o controle das infecções herpéticas vem melhorando cada vez mais. Levando em consideração todos esses fatores, os autores descrevem, nessa revisão de literatura, os principais aspectos relacionados à infecção herpética, quanto às suas características clínicas, diagnóstico diferencial, métodos de diagnóstico e principais tratamentos

Advances in epidemiology, diagnosis and treatment of genital herpes / 中华男科学杂志

Changes in sexual behavior may constitute an important contribution to relative increase in genital herpes. Many research results suggest that HSV-2 plays a big part in the transmission of HIV infection. There is an urgent need to identify effective HSV-2 control measures in order to reduce HIV incidence. The detection of HSV-2-specific antibodies in serum is especially useful and reliable. During the past 2 decades, selective and specific inhibitors of replication have been developed. Such agents as acyclovir, valaciclovir and famciclovir, have reduced the time of healing and the possibility of virus excretion. Efforts have been made in different directions including the exploration of new targets for antiviral chemotherapy, the use of immunomodulators and the development of specific vaccines.