Study on the Immunogenicity of Three Recombinant Lactic Acid Bacteria Expressing ASFV p14.5 Protein

Abstract The African Classical Swine Fever Virus (ASFV) has spread severely all over the world. The lack of vaccines has dealt a heavy blow to the pig industry.In this study, the p14.5 protein encoded by the African swine fever virus was used as the antigen, and the p14.5 protein gene was expressed in vitro using the Lactobacillus expression system. Three new functional recombinant Lactobacillus plantarum((L. plantarum) were constructed and the p14.5 was successfully detected using western technology.Protein, fusion gene p14.5-IL-33-mouse（P14.5-IL-33-Mus） protein and CTA1-p14.5-DD protein expression.After oral immunization of SPF mice with recombinant lactic acid bacteria, flow cytometry and ELISA were used to detect that the differentiation and maturity of T, B, and DC cells of the mice were higher than those of the control group, and specific antibodies were produced. In contrast, the immune effect of the adjuvant group was stronger than that of the single antigen group, and the IL-33 adjuvant effect was stronger than that of the CTA1-DD adjuvant. This study provides effective data support for the prevention of African swine fever virus infection with new lactic acid bacteria preparations, and has certain innovative significance.

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GS-441524 inhibits African swine fever virus infection in vitro

Antiviral Research ◽

10.1016/j.antiviral.2021.105081 ◽

2021 ◽

pp. 105081

Author(s):

Zhao Huang ◽

Lang Gong ◽

Zezhong Zheng ◽

Qi Gao ◽

Xiongnan Chen ◽

...

Keyword(s):

Virus Infection ◽

African Swine Fever Virus ◽

African Swine Fever ◽

Fever Virus

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The Role of Interleukine-10 and Interferon-γ as Potential Markers of the Evolution of African Swine Fever Virus Infection in Wild Boar

Pathogens ◽

10.3390/pathogens10060757 ◽

2021 ◽

Vol 10 (6) ◽

pp. 757

Author(s):

Sandra Barroso-Arévalo ◽

Jose A. Barasona ◽

Estefanía Cadenas-Fernández ◽

José M. Sánchez-Vizcaíno

Keyword(s):

Wild Boar ◽

Vaccine Candidate ◽

African Swine Fever Virus ◽

African Swine Fever ◽

Interferon Γ ◽

Fever Virus ◽

Control Group ◽

Challenge Virus ◽

Ifn Γ

African swine fever virus (ASFv) is one of the most challenging pathogens to affect both domestic and wild pigs. The disease has now spread to Europe and Asia, causing great damage to the pig industry. Although no commercial vaccine with which to control the disease is, as yet, available, some potential vaccine candidates have shown good results in terms of protection. However, little is known about the host immune mechanisms underlying that protection, especially in wild boar, which is the main reservoir of the disease in Europe. Here, we study the role played by two cytokines (IL-10 and IFN-γ) in wild boar orally inoculated with the attenuated vaccine candidate Lv17/WB/Rie1 and challenged with a virulent ASFv genotype II isolate. A group of naïve wild boar challenged with the latter isolate was also established as a control group. Our results showed that both cytokines play a key role in protecting the host against the challenge virus. While high levels of IL-10 in serum may trigger an immune system malfunctioning in challenged animals, the provision of stable levels of this cytokine over time may help to control the disease. This, together with high and timely induction of IFN-γ by the vaccine candidate, could help protect animals from fatal outcomes. Further studies should be conducted in order to support these preliminary results and confirm the role of these two cytokines as potential markers of the evolution of ASFV infection.

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Probiotic Potential of a Novel Vitamin B2-Overproducing Lactobacillus plantarum Strain, HY7715, Isolated from Kimchi

Applied Sciences ◽

10.3390/app11135765 ◽

2021 ◽

Vol 11 (13) ◽

pp. 5765

Author(s):

Joo-Yun Kim ◽

Eun-Jung Choi ◽

Jae-Ho Lee ◽

Myeong-Seok Yoo ◽

Keon Heo ◽

...

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

Lactobacillus Plantarum ◽

Probiotic Strain ◽

Optimal Growth ◽

Control Group ◽

High Survival ◽

Vitamin B2 ◽

Riboflavin Deficiency

Vitamin B2, also known as riboflavin, is essential for maintaining human health. The purpose of this study was to isolate novel lactic acid bacteria that overproduce vitamin B2 and to validate their potential as probiotics. In this study, Lactobacillus plantarum HY7715 (HY7715) was selected among lactic acid bacteria isolated from Kimchi. HY7715 showed a very high riboflavin-producing ability compared to the control strain due to the high expression of ribA, ribB, ribC, ribH, and ribG genes. HY7715 produced 34.5 ± 2.41 mg/L of riboflavin for 24 h without consuming riboflavin in the medium under optimal growth conditions. It was able to produce riboflavin in an in vitro model of the intestinal environment. In addition, when riboflavin deficiency was induced in mice through nutritional restriction, higher levels of riboflavin were detected in plasma and urine in the HY7715 administration group than in the control group. HY7715 showed high survival rate in simulated gastrointestinal conditions and had antibiotic resistance below the cutoff MIC value suggested by the European Food Safety Authority; moreover, it did not cause hemolysis. In conclusion, HY7715 could be considered a beneficial probiotic strain for human and animal applications, suggesting that it could be a new alternative to address riboflavin deficiency.

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Amylase-Producing Maltooligosaccharide Provides Potential Relief in Rats with Loperamide-Induced Constipation

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2020/5470268 ◽

2020 ◽

Vol 2020 ◽

pp. 1-9 ◽

Cited By ~ 1

Author(s):

Eun Yeong Jang ◽

Yejin Ahn ◽

Hyung Joo Suh ◽

Ki-Bae Hong ◽

Kyungae Jo

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

Positive Control ◽

Lactobacillus Bulgaricus ◽

Control Group ◽

Sprague Dawley ◽

Potential Relief ◽

Bowel Movements ◽

Mucosal Thickness

Constipation is a chronic disease caused by infrequent, inadequate, and difficult bowel movements. The present study aimed to evaluate the potential laxative effect of maltooligosaccharide (MOS) on loperamide-induced constipation in a rat model. In vitro experiments were conducted to evaluate the effect of MOS on the growth of lactic acid bacteria. Moreover, to examine the effect of MOS administration on Sprague-Dawley (SD) rats with loperamide-induced constipation, the drinking water for the rats was supplemented with 10% or 15% of MOS for 14 days, and, thereafter, the improvement in constipation was assessed. For this, the rats were divided into five groups: normal (Nor), loperamide-induced constipated (Con), positive control (15% of dual-oligosaccharide (DuO-15)), 10% MOS treated (MOS-10), and 15% MOS-treated (MOS-15). In an in vitro test, MOS treatment promoted the growth of lactic acid bacteria except Lactobacillus bulgaricus. Treatment with higher MOS dose relieved constipation in rats by improving the fecal pellet and water content. Furthermore, in the high MOS dose group, the cecal short-chain fatty acid levels significantly increased compared to those in the control group (P<0.001). MOS treatment also improved the mucosal thickness as well as mucin secretion and increased the area of intestinal Cajal cells compared to that in the control group (P<0.001). These findings suggest that MOS relieves constipation and has beneficial effect on the gastrointestinal tract, and, therefore, it can be used as an ingredient in functional foods for treating constipation or improving intestinal health.

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Role of African swine fever virus (ASFV) proteins EP153R and EP402R in reducing viral persistence and virulence from attenuated BeninΔDP148R

10.1101/2021.04.16.439957 ◽

2021 ◽

Author(s):

Vlad Petrovan ◽

Anusyah Rathakrishnan ◽

Muneeb Islam ◽

Lynnette Goatley ◽

Katy Moffat ◽

...

Keyword(s):

Virus Replication ◽

Vaccine Development ◽

African Swine Fever Virus ◽

Clinical Signs ◽

African Swine Fever ◽

Viral Persistence ◽

Fever Virus ◽

Post Immunization

The limited knowledge on the role of many of the approximately 170 proteins encoded by African swine fever virus restricts progress towards vaccine development. In this study we investigated the effect of deleting combinations of different genes from a previously attenuated virus, BeninΔDP148R on: virus replication in macrophages, virus persistence and clinical signs post immunization, and induction of protection against challenge. Deletion of either EP402R or EP153R genes individually or in combination from BeninΔDP148R did not reduce virus replication in vitro. However, deletion of EP402R dramatically reduced viral persistence in vivo, whilst maintaining high levels of protection against challenge. The additional deletion of EP153R (BeninΔDP148RΔEP153RΔEP402R) further attenuated the virus and no viremia or clinical signs were observed post immunization. This was associated with decreased protection and detection of moderate levels of challenge virus in blood. Interestingly, the deletion of EP153R alone from BeninΔDP148R did not result in further virus attenuation and a slight increase in virus genome copies in blood was observed at different times post immunization when compared with BeninΔDP148R. These results show that EP402R and EP153R have a synergistic role in promoting viremia, however EP153R alone does not seem to have a major impact on virus levels in blood.

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Deletion of a CD2-Like Gene, 8-DR, from African Swine Fever Virus Affects Viral Infection in Domestic Swine

Journal of Virology ◽

10.1128/jvi.72.4.2881-2889.1998 ◽

1998 ◽

Vol 72 (4) ◽

pp. 2881-2889 ◽

Cited By ~ 74

Author(s):

M. V. Borca ◽

C. Carrillo ◽

L. Zsak ◽

W. W. Laegreid ◽

G. F. Kutish ◽

...

Keyword(s):

Viral Infection ◽

Mononuclear Cells ◽

African Swine Fever Virus ◽

Disease Onset ◽

African Swine Fever ◽

Fever Virus ◽

Parental Virus ◽

Viral Virulence

ABSTRACT An African swine fever virus (ASFV) gene with similarity to the T-lymphocyte surface antigen CD2 has been found in the pathogenic African isolate Malawi Lil-20/1 (open reading frame [ORF] 8-DR) and a cell culture-adapted European virus, BA71V (ORF EP402R) and has been shown to be responsible for the hemadsorption phenomenon observed for ASFV-infected cells. The structural and functional similarities of the ASFV gene product to CD2, a cellular protein involved in cell-cell adhesion and T-cell-mediated immune responses, suggested a possible role for this gene in tissue tropism and/or immune evasion in the swine host. In this study, we constructed an ASFV 8-DR gene deletion mutant (Δ8-DR) and its revertant (8-DR.R) from the Malawi Lil-20/1 isolate to examine gene function in vivo. In vitro, Δ8-DR, 8-DR.R, and the parental virus exhibited indistinguishable growth characteristics on primary porcine macrophage cell cultures. In vivo,8-DR had no obvious effect on viral virulence in domestic pigs; disease onset, disease course, and mortality were similar for the mutant Δ8-DR, its revertant 8-DR.R, and the parental virus. Altered viral infection was, however, observed for pigs infected with Δ8-DR. A delay in spread to and/or replication of Δ8-DR in the draining lymph node, a delay in generalization of infection, and a 100- to 1,000-fold reduction in virus titers in lymphoid tissue and bone marrow were observed. Onset of viremia for Δ8-DR-infected animals was significantly delayed (by 2 to 5 days), and mean viremia titers were reduced approximately 10,000-fold at 5 days postinfection and 30- to 100-fold at later times; moreover, unlike in 8-DR.R-infected animals, the viremia was no longer predominantly erythrocyte associated but rather was equally distributed among erythrocyte, leukocyte, and plasma fractions. Mitogen-dependent lymphocyte proliferation of swine peripheral blood mononuclear cells in vitro was reduced by 90 to 95% following infection with 8-DR.R but remained unaltered following infection with Δ8-DR, suggesting that 8-DR has immunosuppressive activity in vitro. Together, these results suggest an immunosuppressive role for 8-DR in the swine host which facilitates early events in viral infection. This may be of most significance for ASFV infection of its highly adapted natural host, the warthog.

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African Swine Fever Virus Infection of Porcine Aortic Endothelial Cells Leads to Inhibition of Inflammatory Responses, Activation of the Thrombotic State, and Apoptosis

Journal of Virology ◽

10.1128/jvi.75.21.10372-10382.2001 ◽

2001 ◽

Vol 75 (21) ◽

pp. 10372-10382 ◽

Cited By ~ 31

Author(s):

Isabelle Vallée ◽

Stephen W. G. Tait ◽

Penelope P. Powell

Keyword(s):

Endothelial Cells ◽

African Swine Fever Virus ◽

African Swine Fever ◽

Surface Expression ◽

Fever Virus ◽

Parp Cleavage ◽

Aortic Endothelial Cells ◽

Porcine Aortic Endothelial Cells ◽

Aortic Endothelial

ABSTRACT African swine fever (ASF) is an asymptomatic infection of warthogs and bushpigs, which has become an emergent disease of domestic pigs, characterized by hemorrhage, lymphopenia, and disseminated intravascular coagulation. It is caused by a large icosohedral double-stranded DNA virus, African swine fever virus (ASFV), with infection of macrophages well characterized in vitro and in vivo. This study shows that virulent isolates of ASFV also infect primary cultures of porcine aortic endothelial cells and bushpig endothelial cells (BPECs) in vitro. Kinetics of early and late gene expression, viral factory formation, replication, and secretion were similar in endothelial cells and macrophages. However, ASFV-infected endothelial cells died by apoptosis, detected morphologically by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling and nuclear condensation and biochemically by poly(ADP-ribose) polymerase (PARP) cleavage at 4 h postinfection (hpi). Immediate-early proinflammatory responses were inhibited, characterized by a lack of E-selectin surface expression and interleukin 6 (IL-6) and IL-8 mRNA synthesis. Moreover, ASFV actively downregulated interferon-induced major histocompatibility complex class I surface expression, a strategy by which viruses evade the immune system. Significantly, Western blot analysis showed that the 65-kDa subunit of the transcription factor NF-κB, a central regulator of the early response to viral infection, decreased by 8 hpi and disappeared by 18 hpi. Both disappearance of NF-κB p65 and cleavage of PARP were reversed by the caspase inhibitor z-VAD-fmk. Interestingly, surface expression and mRNA transcription of tissue factor, an important initiator of the coagulation cascade, increased 4 h after ASFV infection. These data suggest a central role for vascular endothelial cells in the hemorrhagic pathogenesis of the disease. Since BPECs infected with ASFV also undergo apoptosis, resistance of the natural host must involve complex pathological factors other than viral tropism.

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An African swine fever virus ORF with similarity to C-type lectins is non-essential for growth in swine macrophages in vitro and for virus virulence in domestic swine

Journal of General Virology ◽

10.1099/0022-1317-80-10-2693 ◽

1999 ◽

Vol 80 (10) ◽

pp. 2693-2697 ◽

Cited By ~ 28

Author(s):

J. G. Neilan ◽

M. V. Borca ◽

Z. Lu ◽

G. F. Kutish ◽

S. B. Kleiboeker ◽

...

Keyword(s):

African Swine Fever Virus ◽

African Swine Fever ◽

Fever Virus ◽

Virus Infectivity ◽

Mrna Transcription ◽

Host Cell Interactions ◽

Lectin Family ◽

Virus Replication Cycle ◽

Domestic Swine

An African swine fever virus (ASFV) ORF, 8CR, with similarity to the C-type lectin family of adhesion proteins has been described in the pathogenic isolate Malawi Lil-20/1. The similarity of 8CR to cellular and poxvirus genes associated with cell adhesion, cell recognition and virus infectivity suggested that 8CR may be of significance to ASFV–host cell interactions. Sequence analysis of the 8CR ORF from additional pathogenic ASFV isolates demonstrated conservation among isolates from both pig and tick sources. Northern blot analysis demonstrated 8CR mRNA transcription late in the virus replication cycle. A Malawi Lil-20/1 8CR deletion mutant (Δ8CR) was constructed to analyse 8CR function further. The growth characteristics in vitro of Δ8CR in porcine macrophage cell cultures were identical to those observed for parental virus. In domestic swine, Δ8CR exhibited an unaltered parental Malawi Lil- 20/1 disease and virulence phenotype. Thus, although well conserved among pathogenic ASFV field isolates, 8CR is non-essential for growth in porcine macrophages in vitro and for virus virulence in domestic swine.

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