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A study of lymphoid organs and serum proinflammatory cytokines in pigs infected with African swine fever virus genotype II.

Abstract (Expand)

African swine fever virus (ASFV), the causative agent of one of the most important viral diseases of domestic pigs for which no vaccine is available, causes immune system disorders in infected animals. … In this study, the serum levels of proinflammatory cytokines, as well as the histological and cellular constitution of lymphoid organs of pigs infected with ASFV genotype II were investigated. The results showed a high degree of lymphocyte depletion in the lymphoid organs, particularly in the spleen and lymph nodes, where ASFV infection led to a twofold decrease in the number of lymphocytes on the final day of infection. Additionally, ASFV-infected pigs had atypical forms of lymphocytes found in all lymphoid organs. In contrast to lymphocytes, the number of immature immune cells, particularly myelocytes, increased dramatically and reached a maximum on day 7 postinfection. The serum levels of TNF-α, IL-1β, IL-6, and IL-8 were evaluated. Proinflammatory cytokines showed increased levels after ASFV infection, with peak values at 7 days postinfection, and this highlights their role in the pathogenesis of ASFV. In conclusion, this study showed that ASFV genotype II, like other highly virulent strains, causes severe pathological changes in the immune system of pigs.

Authors: Hovakim Zakaryan, Victorya Cholakyans, Lusine Simonyan, Alla Misakyan, Elena Karalova, Andranik Chavushyan, Zaven Karalyan

Date Published: 24th Mar 2015

Publication Type: Journal

Evaluation of hemostaseological status of pigs experimentally infected with African swine fever virus.

Abstract (Expand)

African swine fever is a highly contagious hemorrhagic disease of pigs caused by African swine fever virus (ASFV). Hemorrhages are the most frequently reported lesions in acute and subacute forms of … ASF. Hemorrhagic lesions are accompanied by impaired hemostasis, which includes thrombocytopenia and changes in the coagulation system. In the present study, experimental infection was conducted to elucidate whether a highly virulent ASFV genotype II circulating in the Trans-Caucasus and Eastern Europe affects the hemostasis of infected pigs. Platelet count changes and platelet size, as well as coagulation parameters were evaluated upon experimental infection. In contrast to other ASFV strains, ASFV genotype II showed a significant decrease in the number of platelets from 3rd dpi onwards. Furthermore, a decrease in platelet size was observed throughout the entire period of experiment. A significant increase in the number of platelet aggregates was observed from the beginning of infection. Unlike other ASFV strains, ASFV genotype II induced a slight shortening of an activated partial thromboplastin time (aPTT) throughout the experiment. Thrombin time (TT) was prolonged from day 5 onwards, whereas no changes in prothrombin time (PT) were found upon infection. The level of d-dimers was permanently higher than in control with a peak on day 3 post-infection. ASFV induced a significant decrease in the level of fibrinogen from day 5 till the end of experiment. Thus, it can be concluded that ASFV genotype II isolated in Armenia affects the hemostasis of infected pigs and causes changes that differ from that of other ASFV strains described previously.

Authors: Hovakim Zakaryan, Elena Karalova, Henrik Voskanyan, Zarine Ter-Pogossyan, Narek Nersisyan, Astghik Hakobyan, David Saroyan, Zaven Karalyan

Date Published: 7th Nov 2014

Publication Type: Journal

Pathology of porcine peripheral white blood cells during infection with African swine fever virus.

Abstract (Expand)

African swine fever virus (ASFV) is the causative agent of African swine fever (ASF) that is the significant disease of domestic pigs. Several studies showed that ASFV can influence on porcine blood … cells in vitro. Thus, we asked ourselves whether ASFV infection results in changes in porcine blood cells in vivo. A series of experiments were performed in order to investigate the effects of ASFV infection on porcine peripheral white blood cells. Nine pigs were inoculated by intramuscular injection with 10⁴ 50% hemadsorbing doses of virus (genotype II) distributed in Armenia and Georgia. The total number of fifteen cell types was calculated during experimental infection.

Authors: Zaven Karalyan, Hovakim Zakaryan, Hranush Arzumanyan, Khachik Sargsyan, Henrik Voskanyan, Lina Hakobyan, Liana Abroyan, Aida Avetisyan, Elena Karalova

Date Published: 28th Feb 2012

Publication Type: Journal

Interferon status and white blood cells during infection with African swine fever virus in vivo.

Abstract (Expand)

African swine fever virus (ASFV) is the causative agent of African swine fever that is the significant disease of domestic pigs, with high rates of mortality. ASFV is double-stranded DNA virus whose … genes encode some proteins that are implicated in the suppression of host immune response. In this study, we have modeled in vivo infection of ASFV for determination of interferon (IFN) status in infected pigs. We measured the level of IFN-α, -β and -γ by enzyme-linked immunosorbent assay and showed that the level of IFN-α sharply decreased during infection. Unlike IFN-α, the level of IFN-β and -γ increased from the 2nd and 4th days post-infection, respectively. Also, we analyzed the population dynamics of peripheral white blood cells of infected pigs due to their important role in host immune system. We showed that the atypical lymphocytes appeared after short time of infection and this result is in accordance with our previous study done in vitro. At the last day of infection about 50% of the total white blood cells were destroyed, and the remaining cells were represented mainly by small-sized lymphocytes, reactive lymphocytes and lymphoblasts.

Authors: Z Karalyan, H Zakaryan, Kh Sargsyan, H Voskanyan, H Arzumanyan, H Avagyan, E Karalova

Date Published: 15th Jan 2012

Publication Type: Journal

Phenotypic and cytologic studies of lymphoid cells and monocytes in primary culture of porcine bone marrow during infection of African swine fever virus.

Abstract (Expand)

We have modeled in vitro infection of African swine fever virus (ASFV) in primary unstimulated cells of the porcine bone marrow and have studied the phenotypical changes in the population of porcine … lymphoid cells by cytophotometry. Monocytes and large-sized lymphocytes completely vanished in 72 h of infection which is result of high sensitivity of those cells to ASFV. We describe DNA synthesis in monocytes at 24 h post infection. Cytophotometry of the uninfected cells revealed the few number of atypical lymphocytes and lymphoblasts after 72 h of cultivation; whereas in viral infected cultures, atypical cells appeared in large quantity (about 14%) with 24 h. Most of atypical lymphocytes and lymphoblasts had altered nucleus, and only a small number of atypical cells had additional nucleus. The cytophotometry of main and additional nuclei showed that DNA content didn't exceed diploid standard which indicates that the additional nuclei were consequence of fragmentation of nuclei in lymphocytes.

Authors: E M Karalova, Kh V Sargsyan, G K Hampikian, H E Voskanyan, L O Abroyan, A S Avetisyan, L A Hakobyan, H H Arzumanyan, H S Zakaryan, Zaven A Karalyan

Date Published: 24th Dec 2010

Publication Type: Journal

A new microtubule-stabilizing agent shows potent antiviral effects against African swine fever virus with no cytotoxicity.

Abstract (Expand)

African swine fever virus (ASFV) is the causal agent of a fatal disease of domestic swine for which no effective antiviral drugs are available. Recently, it has been shown that microtubule-targeting … agents hamper the infection cycle of different viruses. In this study, we conducted in silico screening against the colchicine binding site (CBS) of tubulin and found three new compounds with anti-ASFV activity. The most promising antiviral compound (6b) reduced ASFV replication in a dose-dependent manner (IC50 = 19.5 μM) with no cellular (CC50 > 500 μM) and animal toxicity (up to 100 mg/kg). Results also revealed that compound 6b interfered with ASFV attachment, internalization and egress, with time-of-addition assays, showing that compound 6b has higher antiviral effects when added within 2-8 h post-infection. This compound significantly inhibited viral DNA replication and disrupted viral protein synthesis. Experiments with ASFV-infected porcine macrophages disclosed that antiviral effects of the compound 6b were similar to its effects in Vero cells. Tubulin polymerization assay and confocal microscopy demonstrated that compound 6b promoted tubulin polymerization, acting as a microtubule-stabilizing, rather than a destabilizing agent in cells. In conclusion, this work emphasizes the idea that microtubules can be targets for drug development against ASFV.

Authors: Samvel Sirakanyan, Erik Arabyan, Astghik Hakobyan, Tamara Hakobyan, Garri Chilingaryan, Harutyun Sahakyan, Arsen Sargsyan, Grigor Arakelov, Karen Nazaryan, Roza Izmailyan, Liana Abroyan, Zaven Karalyan, Elina Arakelova, Elmira Hakobyan, Anush Hovakimyan, Andre Serobian, Marco Neves, João Ferreira, Fernando Ferreira, Hovakim Zakaryan

Date Published: No date defined

Publication Type: Journal

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