Equine Herpesvirus Type 1 Enhances Viral Replication in CD172a + Monocytic Cells upon Adhesion to Endothelial Cells
Open accessAbstract
UNLABELLED: Equine herpesvirus type 1 (EHV-1) is a main cause of respiratory disease, abortion, and encephalomyelopathy in horses. Monocytic cells (CD172a(+)) are the main carrier cells of EHV-1 during primary infection and are proposed to serve as a "Trojan horse" to facilitate the dissemination of EHV-1 to target organs. However, the mechanism by which EHV-1 is transferred from CD172a(+) cells to endothelial cells (EC) remains unclear. The aim of this study was to investigate EHV-1 transmission between these two cell types. We hypothesized that EHV-1 employs specific strategies to promote the adhesion of infected CD172a(+) cells to EC to facilitate EHV-1 spread. Here, we demonstrated that EHV-1 infection of CD172a(+) cells resulted in a 3- to 5-fold increase in adhesion to EC. Antibody blocking experiments indicated that α4β1, αLβ2, and αVβ3 integrins mediated adhesion of infected CD172a(+) cells to EC. We showed that integrin-mediated phosphatidylinositol 3-kinase (PI3K) and ERK/MAPK signaling pathways were involved in EHV-1-induced CD172a(+) cell adhesion at early times of infection. EHV-1 replication was enhanced in adherent CD172a(+) cells, which correlates with the production of tumor necrosis factor alpha (TNF-α). In the presence of neutralizing antibodies, approximately 20% of infected CD172a(+) cells transferred cytoplasmic material to uninfected EC and 0.01% of infected CD172a(+) cells transmitted infectious virus to neighboring cells. Our results demonstrated that EHV-1 infection induces adhesion of CD172a(+) cells to EC, which enhances viral replication, but that transfer of viral material from CD172a(+) cells to EC is a very specific and rare event. These findings give new insights into the complex pathogenesis of EHV-1. IMPORTANCE: Equine herpesvirus type 1 (EHV-1) is a highly prevalent pathogen worldwide, causing frequent outbreaks of abortion and myeloencephalopathy, even in vaccinated horses. After primary replication in the respiratory tract, EHV-1 disseminates via cell-associated viremia in peripheral blood mononuclear cells (PBMC) and subsequently infects the endothelial cells (EC) of the pregnant uterus or central nervous system, leading in some cases to abortion and/or neurological disorders. Recently, we demonstrated that CD172a(+) monocytic carrier cells serve as a "Trojan horse" to facilitate EHV-1 spread from blood to target organs. Here, we investigated the mechanism underlying the transmission of EHV-1 from CD172a(+) cells to EC. We demonstrated that EHV-1 infection induces cellular changes in CD172a(+) cells, promoting their adhesion to EC. We found that both cell-to-cell contacts and the secretion of soluble factors by EC activate EHV-1 replication in CD172a(+) cells. This facilitates transfer of cytoplasmic viral material to EC, resulting mainly in a nonproductive infection. Our findings give new insights into how EHV-1 may spread to EC of target organs in vaccinated horses.
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- Findingown data§ RESULTS · p. 5
Phosphorylated ERK1/2 and Akt were not detected in CD172a+ cells even with a sensitive assay.
“However, we failed to detect any levels of ERK1/2 and Akt phosphorylation, even by using a highly sensitive detection kit (Fig. 3A).”
Quote in the paperNot a substantial claim - Findingown data§ ABSTRACT · p. 1
EHV-1 infection raises the adhesion of CD172a+ monocytic cells to equine endothelial cells by roughly three- to fivefold.
“Here, we demonstrated that EHV-1 infection of CD172a ÷ cells resulted in a 3- to 5-fold increase in adhesion to EC.”
- Also stated in: § RESULTS · p. 4 · plain
“EHV-1 infection resulted in a significant increase ( P < 0.001) in adhesion of CD172a + cells to primary and immortalized venous EC monolayers that was independent of the EHV-1 strain used (03P37 or 97P70) (Fig. 1A, B, and C).”
- Also stated in: § RESULTS · p. 4 · plain
“A maximum of 200 to 230 adherent cells per mm 2 was reached with EHV-1-inoculated CD172a + cells after 2 h of coculture compared to approximately 50 adherent cells per mm 2 with mock-inoculated CD172a + cells.”
Quote in the paperSubstantial claim - Also stated in: § RESULTS · p. 4 · plain
- Findingown data§ ABSTRACT · p. 1
With neutralizing antibodies present, about 0.01% of infected CD172a+ cells pass infectious virus to neighboring cells.
“and 0.01% of infected CD172a ÷ cells transmitted infectious virus to neighboring cells.”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 5
After integrin blockade, adhesion of EHV-1-inoculated CD172a+ cells falls to about the mock level or slightly below, around 50 cells per mm2.
“After preincubation with blocking antibodies, the number of adherent CD172a + cells per square millimeter in EHV-1-inoculated cells decreased to levels at or slightly below those of mockinoculated cells ( ± 50 adherent cells per mm 2 ).”
Quote in the paperSubstantial claim - Findingown data§ ABSTRACT · p. 1
With neutralizing antibodies present, about 20% of infected CD172a+ cells transfer cytoplasmic material to uninfected endothelial cells.
“In the presence of neutralizing antibodies, approximately 20% of infected CD172a ÷ cells transferred cytoplasmic material to uninfected EC”
Quote in the paperSubstantial claim
- Findingown data§ RESULTS · p. 6
With 10 μM U0126, EHV-1-induced adhesion of CD172a+ cells to endothelial cells drops by 55% for strain 03P37 and 61% for strain 97P70.
“When CD172a + cells were pretreated with 10 𝛍 M U0126, EHV-1 (03P37 and 97P70)-induced CD172a + cell adhesion to EC monolayers was reduced by 55 and 61%, respectively.”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 6
Blocking ERK/MAPK signaling before EHV-1 inoculation reduces CD172a+ cell adhesion to endothelial cells in a dose-dependent way, down to mock levels.
“Inhibition of the ERK/MAPK signaling pathway prior to EHV-1 inoculation resulted in a dose-dependent significant decrease ( P < 0.001) in CD172a + cell adhesion to mocklevels (Fig. 3B and D).”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 6
U0126 or LY294002 treatment does not significantly change adhesion of mock-inoculated CD172a+ cells.
“Pretreating mock-inoculated cells with the inhibitor U0126 or LY294002 had no significant effect on CD172a + cell adhesion.”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 6
Inhibiting ERK/MAPK and PI3K does not significantly reduce EHV-1 infection of CD172a+ cells relative to controls.
“Furthermore, we found that inhibition of the ERK/MAPK and PI3K pathways did not significantly decrease EHV-1 infection of CD172a + cells compared to controls (Fig. 3E).”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 4
The adhesion increase seen with EHV-1-inoculated CD172a+ cells is not specific to the viral strain, occurring with both 03P37 and 97P70.
“EHV-1 infection resulted in a significant increase ( P < 0.001) in adhesion of CD172a + cells to primary and immortalized venous EC monolayers that was independent of the EHV-1 strain used (03P37 or 97P70) (Fig. 1A, B, and C).”
Quote in the paperNot a substantial claim - Findingown data§ RESULTS · p. 4
Primary endothelial cells and immortalized endothelial cell lines support similar adhesion kinetics of CD172a+ cells.
“As the kinetics of CD172a + cell adhesion were similar in primary EC cultures and immortalized EC lines, the subsequent experiments were performed using immortalized EC only.”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 5
Combining blocking antibodies against the three integrins does not reduce CD172a+ cell adhesion more than a single blocking antibody.
“Pretreating cells with a mixture of blocking antibodies had no cumulative effect on decreasing CD172a + cell adhesion to EC compared to treatment of cells with a single antibody.”
Quote in the paperNot a substantial claim - Findingown data§ RESULTS · p. 5
Integrin-blocking antibodies do not significantly reduce adhesion of mock-inoculated CD172a+ cells.
“Interestingly, no significant decrease in cell adhesion was observed with mock-inoculated CD172a + cells.”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 4
UV-inactivated EHV-1 and replication-competent EHV-1 produce comparable adhesion kinetics for CD172a+ cells on endothelial monolayers.
“In addition, no significant difference in the kinetics of adhesion of CD172a + cells to EC monolayers was observed between UV- EHV-1- and EHV-1-treated cells.”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 6
Blocking PI3K before EHV-1 inoculation produces a comparable reduction in CD172a+ cell adhesion to endothelial cells.
“Inhibition of PI3K activity prior to EHV-1 inoculation resulted in a similar decrease of CD172a + cell adhesion (Fig. 3C).”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 5
Total ERK1/2 and total Akt levels in CD172a+ cells are similar between mock and EHV-1 inoculation at 0, 5, and 15 minutes after inoculation.
“We found that mock- and EHV-1-inoculated CD172a + cells showed similar levels of total ERK1/2 and total Akt expression at 0, 5, and 15 min postinoculation.”
Quote in the paperNot a substantial claim - Findingown data§ RESULTS · p. 7
The percentage of IEP-positive cells decreased in the nonadherent CD172a+ cell fraction over time postadhesion for both EHV-1 strains.
“The percentage of IEP-positive cells decreased in the nonadherent CD172a + cell fraction over time postadhesion for both EHV-1 strains (Fig. 4B).”
Quote in the paperNot a substantial claim - Findingown data§ RESULTS · p. 8
IEP/gB-positive plaques could not be observed in EC at later time points postinoculation, indicating that spread of infection from CD172a+ cells to EC did not automatically lead to productive infection in EC.
“However, IEP/gB-positive plaques also could not be observed in EC at later time points postinoculation, indicating that spread of infection from CD172a + cells to EC did not automatically lead to productive infection in EC.”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 8
Addition of neutralizing TNFα antibody to cocultures of CD172a+ cells adherent to EC generated a trend, not statistically significant, toward a decrease in the percentage of gB-positive cells.
“Moreover, we demonstrated that the addition of neutralizing TNF𝛂 antibody to cocultures of CD172a + cells adherent to EC generated a trend, although not statistically significant, toward a decrease in the percentage of gB-positive cells (Fig. 5C).”
Quote in the paperSubstantial claim - Findingown data§ DISCUSSION · p. 10
Approximately 20% of late infected CD172a+ cells were able to transfer cytoplasmic EHV-1 material to uninfected, adjacent EC in the presence of neutralizing antibodies.
“Finally, we demonstrated that approximately 20% of late infected CD172a + cells were able to transfer cytoplasmic EHV-1 material to uninfected, adjacent EC in the presence of neutralizing antibodies (Fig. 7D).”
Quote in the paperSubstantial claim - Findingown data§ DISCUSSION · p. 10
Transfer of EHV-1 from late infected CD172a+ cells to EC predominantly resulted in a nonproductive infection in EC.
“In our experiments, transfer of EHV-1 from late infected CD172a + cells to EC predominantly resulted in a nonproductive infection in EC.”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 7
No significant difference in TNFα concentrations between supernatants of mock-inoculated CD172a+ cells cultured on plastic or on top of EC.
“No significant difference was detected in TNF𝛂 concentrations between supernatants of mock-inoculated CD172a + cells cultured on plastic or on top of EC.”
Quote in the paperNot a substantial claim - Findingown data§ DISCUSSION · p. 9
Blocking α4β1 (VLA-4), αLβ2 (LFA-1), and αVβ3 integrins on EHV-1-inoculated CD172a+ cells significantly reduced adhesion to levels comparable to mock-inoculated cells.
“Here, we showed that blocking 𝛂 4 𝛃 1 (VLA-4), 𝛂 L 𝛃 2 (LFA-1), and 𝛂 V 𝛃 3 integrins on EHV-1-inoculated CD172a + cells significantly reduced adhesion to levels comparable to those of mock-inoculated cells.”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 7
No gB protein was detected in cells from 2 to 6 h postadhesion.
“No gB protein was detected in cells from 2 to 6 h p.a.”
Quote in the paperNot a substantial claim - Findingown data§ RESULTS · p. 7
The expression levels of gB in IEP-positive CD172a+ cells upon adhesion to EC were comparable between the two strains (60%).
“The expression levels of gB in IEP-positive CD172a + cells upon adhesion to EC were comparable between the two strains (60%).”
Quote in the paperNot a substantial claim - Findingown data§ DISCUSSION · p. 9
UV treatment of EHV-1 did not affect the EHV-1-triggered adhesion process.
“UV treatment of EHV-1 did not affect the EHV-1-triggered adhesion process, indicating that the binding of EHV-1toCD172a + cells was sufficient to induce their adhesion to EC.”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 8
Some cytoplasmic IEP staining was observed in the adjacent EC.
“Some cytoplasmic IEP staining was also observed in the adjacent EC.”
Quote in the paperNot a substantial claim - Findingown data§ DISCUSSION · p. 9
EHV-1 infection increased adhesion of CD172a+ monocytic cells to EC in vitro.
“In the present study, we demonstrated that EHV-1 infection increased adhesion of CD172a + monocytic cells to EC in vitro .”
Quote in the paperSubstantial claim - Findingown data§ DISCUSSION · p. 10
Substantial amounts of interleukin 1β (IL-1β) and alpha interferon (IFNα) were not detected upon coculture of mock- or EHV-1-inoculated CD172a+ cells with ECs.
“However, we failed to detect substantial amounts of interleukin 1 𝛃 (IL-1 𝛃 ) and alpha interferon (IFN𝛂 ) produced upon coculture of mock- or EHV-1-inoculated CD172a + cells with ECs (data not shown).”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 7
For EHV-1 strain 03P37, the percentage of IEP-positive CD172a+ cells increased significantly from 4% at 2 h to 14% at 12 h postadhesion on EC monolayers.
“From 2 to 12 h postadhesion (p.a.) on EC monolayers, the percentage of IEP-positive CD172a + cells significantly increased from 4% to 14% for EHV-1 strain 03P37 ( P < 0.01).”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 7
At 12 h postadhesion, the percentage of IEP-positive CD172a+ cells was significantly higher in cells adherent to EC than in cells adherent to plastic for strain 03P37.
“At 12 h p.a., the percentage of IEP-positive CD172a + cells was significantly higher in cells adherent to EC than in cells adherent to plastic ( P < 0.01) (Fig. 4A, left).”
Quote in the paperSubstantial claim - Findingown data§ DISCUSSION · p. 9
The percentages of IEP- and gB-positive CD172a+ cells were higher in cells adherent to EC than in cells adherent to plastic.
“The percentages of IEPand gB-positive CD172a + cells were higher in cells adherent to EC than in cells adherent to plastic.”
Quote in the paperSubstantial claim - Findingown data§ DISCUSSION · p. 9
Significantly more EHV-1-inoculated CD172a+ cells adhered to EC than mock-inoculated cells.
“Significantly more EHV-1-inoculated CD172a + cells adhered to EC than to mock-inoculated cells.”
Quote in the paperSubstantial claim - Findingown data§ DISCUSSION · p. 9
EHV-1 infection induces CD172a+ cell adhesion in PI3K- and ERK/MAPK-dependent processes at a very early stage of infection.
“We demonstrated that EHV-1 infection induces CD172a + cell adhesion in PI3K- and ERK/MAPK-dependent processes at a very early stage of infection.”
Quote in the paperSubstantial claim - Findingown data§ DISCUSSION · p. 9
A greater increase in gB protein expression was observed in IEP-positive cells inoculated with EHV-1 strain 97P70 and cocultured for 12 h with EC monolayers than in cells inoculated with EHV-1 03P37.
“Moreover, we observed a greater increase in gBprotein expression in IEP-positive cells inoculated with EHV-1 strain 97P70 and cocultured for 12 h with EC monolayers than in cells inoculated with EHV-1 03P37.”
Quote in the paperSubstantial claim - Findingown data§ DISCUSSION · p. 10
EHV-1 neurovirulent and nonneurovirulent strains both enhanced EHV-1 replication in CD172a+ cells upon adhesion to EC.
“Interestingly, EHV-1 neurovirulent and nonneurovirulent strains both enhanced EHV-1 replication in CD172a + cells upon adhesion to EC.”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 7
No significant difference in the percentage of IEP-positive cells was observed in CD172a+ cells inoculated with EHV-1 strain 03P37 or 97P70 and cultured on plastic.
“No significant difference in the percentage of IEP-positive cells was observed in CD172a + cells inoculated with EHV-1 strain 03P37 or 97P70 and cultured on plastic.”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 8
IEP was expressed in nuclear replicative compartments and in the cytoplasm of infected CD172a+ cells adherent to EC.
“We found that IEP was expressed in nuclear replicative compartments and in the cytoplasm of infected CD172a + cells adherent to EC.”
Quote in the paperNot a substantial claim - Findingown data§ RESULTS · p. 8
18% and 27% of IEP/gB-positive CD172a+ cells inoculated with EHV-1 strains 03P37 and 97P70, respectively, showed cytoplasmic transfer of EHV-1 material to adjacent EC.
“Eighteen and 27% of IEP/gB (03P37 and 97P70)-positive CD172a + cells showed cytoplasmic transfer of EHV-1 material to adjacent EC.”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 8
With both EHV-1 strains, a cytoplasmic IEP-positive bridge was formed from an infected CD172a+ cell to an adjacent EC.
“With both EHV-1 strains, a cytoplasmic IEP-positive 'bridge' was formed from an infected CD172a + cell to an adjacent EC (Fig. 6B).”
Quote in the paperSubstantial claim - Findingown data§ DISCUSSION · p. 10
The percentage of gB-positive CD172a+ cells adherent to EC was reduced following treatment with TNFα-neutralizing antibody.
“We also demonstrated that the percentage of gB-positive CD172a + cells adherent to EC was reduced following treatment with TNF𝛂 -neutralizing antibody.”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 8
After 5 days of coculture of inoculated CD172a+ cells with EC monolayers, 0.1% and 0.05% of CD172a+ cells inoculated with EHV-1 strains 03P37 and 97P70, respectively, transmitted the infection to EC.
“After 5 days of coculture of inoculated CD172a + cells with EC monolayers, we showed that 0.1% and 0.05% of CD172a + cells inoculated with EHV-1 strains 03P37 and 97P70, respectively, transmitted the infection to EC.”
Quote in the paperSubstantial claim - Findingown data§ DISCUSSION · p. 9
The specific MEK/ERK inhibitor U0126 and the PI3K inhibitor LY294002 efficiently decreased CD172a+ cell adhesion to EC in a dose-dependent manner.
“The specific MEK/ERK inhibitor U0126 and the PI3K inhibitor LY294002 were able to efficiently decrease CD172a + cell adhesion to EC in a dose-dependent manner.”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 7
At 12 h postadhesion, TNFα concentration in supernatants of CD172a+ cells inoculated with EHV-1 strain 03P37 and cocultured with EC was significantly higher (P<0.05) than in EHV-1-inoculated cells cultured on plastic.
“At 12 h p.a., the concentration of TNF𝛂 in supernatants of CD172a + cells inoculated with EHV-1 strain 03P37 or 97P70 and cocultured with EC was significantly higher ( P < 0.05 and P < 0.001, respectively) than the TNF𝛂 concentration in supernatants of EHV-1-inoculated cells cultured on plastic (Fig. 5A).”
Quote in the paperNot a substantial claim - Findingown data§ RESULTS · p. 7
At 12 h postadhesion, TNFα concentration in supernatants of CD172a+ cells inoculated with EHV-1 strain 97P70 and cocultured with EC was significantly higher (P<0.001) than in EHV-1-inoculated cells cultured on plastic.
“At 12 h p.a., the concentration of TNF𝛂 in supernatants of CD172a + cells inoculated with EHV-1 strain 03P37 or 97P70 and cocultured with EC was significantly higher ( P < 0.05 and P < 0.001, respectively) than the TNF𝛂 concentration in supernatants of EHV-1-inoculated cells cultured on plastic (Fig. 5A).”
Quote in the paperNot a substantial claim - Findingown data§ RESULTS · p. 8
The late protein gB expressed a weak signal colocalized with IEP in some regions of both the cytoplasmic bridge and the cytoplasm of the EC.
“The late protein gB expressed a weak signal colocalized with IEP in some regions of both the cytoplasmic bridge and the cytoplasm of the EC.”
Quote in the paperSubstantial claim - Findingown data§ RESULTS · p. 6
EHV-1 replication is enhanced in CD172a+ cells upon adhesion to endothelial cells.
“EHV-1replication is enhanced in CD172a ÷ cells upon adhesion to EC.”
- Also stated in: § DISCUSSION · p. 9 · boosted
“we showed that EHV-1 replication was enhanced in CD172a + cells upon adhesion to EC.”
Quote in the paperSubstantial claim- The authors state this as an assumption: Monocytic cells play a key role in EHV-1 spread from the blood to target organs.
- Also stated in: § DISCUSSION · p. 9 · boosted
- Interpretationown data§ RESULTS · p. 5
EHV-1-induced adhesion of CD172a+ cells to endothelial cells requires integrin-mediated PI3K and ERK/MAPK signaling.
“EHV-1-induced adhesion of CD172a ÷ cells to EC is dependent on integrin-mediated PI3K and ERK/MAPK signaling pathways.”
Quote in the paperSubstantial claim - Interpretationown data§ RESULTS · p. 5
Integrins account for part of the interaction between EHV-1-inoculated CD172a+ cells and endothelial cells.
“Taken together, these results show that interactions between EHV-1-inoculated CD172a + cells and EC are in part mediated by integrins.”
Quote in the paperSubstantial claim - Interpretationown data§ IMPORTANCE · p. 1
The findings suggest EHV-1 may spread to endothelial cells of target organs in vaccinated horses.
“Our findings give new insights into how EHV-1 may spread to EC of target organs in vaccinated horses.”
Quote in the paperSubstantial claim- Rests on an assumption: The in vitro coculture model with immortalized endothelial cells reflects the relevant CD172a+ cell-endothelial cell interactions that occur in vivo.
- Interpretationown data§ RESULTS · p. 6
EHV-1 infection drives CD172a+ cell adhesion through PI3K- and ERK/MAPK-dependent mechanisms very early after infection.
“Taken together, these data suggest that EHV-1 infection induces CD172a + cell adhesion in a PI3K- and ERK/ MAPK-dependent manner at a very early time of infection.”
Quote in the paperSubstantial claim - Interpretationown data§ IMPORTANCE · p. 1
Transfer of cytoplasmic viral material to endothelial cells leads mainly to a nonproductive infection.
“This facilitates transfer of cytoplasmic viral material to EC, resulting mainly in a nonproductive infection.”
Quote in the paperSubstantial claim - Interpretationown data§ RESULTS · p. 4
EHV-1-mediated adhesion of CD172a+ cells to endothelial cells depends on cellular adhesion molecules.
“EHV-1-mediated CD172a ÷ cell adhesion to EC depends on cellular adhesion molecules.”
Quote in the paperSubstantial claim - Premisecitation§ IMPORTANCE · p. 1
CD172a+ monocytic cells are the principal carrier cell type for EHV-1 during primary infection.
“Monocytic cells (CD172a + ) have been shown to be the main carrier cell type of EHV-1 during primary infection (8, 9).”
Quote in the paperSubstantial claim0 of 2 sources checked- cites [8] 8. Van der Meulen KM, Nauwynck HJ, Buddaert W, Pensaert MB. 2000. Replication of equine herpesvirus type 1 in freshly isolated equine peripheral blood mononuclear cells and changes in susceptibility following mitogen stimulation. J Gen Virol 81: 21-25. http://dx.doi.org/10.1099/0022 -1317-81-1-21. source
- cites [9] 9. Gryspeerdt AC, Vandekerckhove AP, Baghi HB, Van de Walle GR, Nauwynck HJ. 2012. Expression of late viral proteins is restricted in nasal mucosal leucocytes but not in epithelial cells during early-stage equine herpes virus-1 infection. Vet J 193: 576-578. http://dx.doi.org/10.1016/j .tvjl.2012.01.022. source
- Interpretationown data§ DISCUSSION · p. 11
EHV-1 uses specific cellular signaling pathways to trigger cellular changes in CD172a+ cells, which result in increased adhesion to EC.
“In conclusion, our data suggest that EHV-1 uses specific cellular signaling pathways to trigger cellular changes in CD172a + cells, which result in increased adhesion to EC.”
Quote in the paperSubstantial claim - Interpretationreasoning§ RESULTS · p. 7
Direct cell-to-cell contacts provide a favorable environment for EHV-1 replication in CD172a+ cells.
“Moreover, the enhancement of EHV-1 replication in CD172a + cells upon adhesion to EC suggests that direct cell-to-cell contacts provide a favorable environment for EHV-1 replication in CD172a + cells.”
Quote in the paperSubstantial claim - Interpretationcitation§ DISCUSSION · p. 10
Adhesion and transfer of EHV-1 from CD172a+ monocytic cells to endothelial cells may contribute to clinical thrombosis in horses with EHV-1 infection.
“Based on this, we believe that the adhesion and transfer of EHV-1 from CD172a + monocytic cells to endothelial cells may contribute to clinical thrombosis in horses with EHV-1 infection.”
Quote in the paperSubstantial claim0 of 3 sources checked- cites [47] 47. Edington N, Bridges CG, Patel JR. 1986. Endothelial cell infection and thrombosis in paralysis caused by equid herpesvirus-1: equine stroke. Arch Virol 90: 111-124. http://dx.doi.org/10.1007/BF01314149. source
- cites [48] 48. Patel JR, Edington N, Mumford JA. 1982. Variation in cellular tropism between isolates of equine herpesvirus-1 in foals. Arch Virol 74: 41-51. http://dx.doi.org/10.1007/BF01320781. source
- cites [49] 49. Smith KC, Whitwell KE, Binns MM, Dolby CA, Hannant D, Mumford JA. 1992. Abortion of virologically negative foetuses following experimental challenge of pregnant pony mares with equid herpesvirus 1. Equine Vet J 24: 256-259. http://dx.doi.org/10.1111/j.2042-3306.1992.tb02830.x. source
- Interpretationown data§ RESULTS · p. 8
Direct cytoplasmic transfer of EHV-1 material from late infected CD172a+ cells to adjacent EC typically leads to a nonproductive infection in these cells.
“Taken together, these results provide evidence for direct cytoplasmic transfer of EHV-1 material from late infected CD172a + cells to adjacent EC, which typically leads to a nonproductive infection in these cells.”
Quote in the paperSubstantial claim - Interpretationown data§ RESULTS · p. 8
TNFα is an important soluble mediator enhancing EHV-1 replication in CD172a+ cells upon adhesion to EC.
“Taken together, these results indicate that TNF𝛂 is an important soluble mediator enhancing EHV-1 replication in CD172a + cells upon adhesion to EC.”
Quote in the paperSubstantial claim - Interpretationown data§ DISCUSSION · p. 9
Binding of EHV-1 to CD172a+ cells is sufficient to induce their adhesion to EC.
“UV treatment of EHV-1 did not affect the EHV-1-triggered adhesion process, indicating that the binding of EHV-1toCD172a + cells was sufficient to induce their adhesion to EC.”
Quote in the paperSubstantial claim - Interpretationown data§ DISCUSSION · p. 9
Integrins mediated the adhesion of CD172a+ cells to EC.
“These results indicated that integrins mediated the adhesion of CD172a + cells to EC.”
Quote in the paperSubstantial claim - Interpretationown data§ DISCUSSION · p. 10
EHV-1 infection of EC does not automatically result in viral spread to the surrounding tissues.
“These data suggest that (i) only a minority of infected CD172a + cells can efficiently transfer EHV-1 to EC and initiate an infection and (ii) EHV-1 infection of EC does not automatically result in viral spread to the surrounding tissues.”
Quote in the paperSubstantial claim - Interpretationcitation§ DISCUSSION · p. 9
The differences in replication kinetics between EHV-1 strains 03P37 and 97P70 in target monocytic cells are due to differences in the replication kinetics of EHV-1 in those cells.
“Taken together, we believe that these results are due to differences in the replication kinetics of EHV-1 in target monocytic cells between the two strains.”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [10] 10. Laval K, Favoreel HW, Nauwynck HJ. 2015. Equine herpesvirus type 1 (EHV-1) replication is delayed in CD172a + monocytic cells and con- source
- Interpretationreasoning§ DISCUSSION · p. 10
Additional factors account for the difference in pathogenesis of EHV-1 in vivo.
“These findings suggest that additional factors account for the difference in pathogenesis of EHV-1 in vivo .”
Quote in the paperNot a substantial claim - Study design and conductnone given§ DISCUSSION · p. 9
The antibodies used in this study do not discriminate between inactive and activated forms of the integrins.
“Indeed, the antibodies used in this study do not discriminate between inactive and activated forms of the integrins, and it was technically impossible to assess any potential upregulation of integrin expression on EHV-1-inoculated cells compared to mock-inoculated cells, due to a low percentage of infected CD172a + cells detected.”
Quote in the paperSubstantial claim - Interpretationown data§ DISCUSSION · p. 10
Only a minority of infected CD172a+ cells can efficiently transfer EHV-1 to EC and initiate an infection.
“These data suggest that (i) only a minority of infected CD172a + cells can efficiently transfer EHV-1 to EC and initiate an infection and (ii) EHV-1 infection of EC does not automatically result in viral spread to the surrounding tissues.”
Quote in the paperSubstantial claim - Interpretationown data§ DISCUSSION · p. 9
Integrin engagement and activation of integrin-mediated signaling pathways are essential in the induction of EHV-1-infected CD172a+ adhesion to EC.
“Together, these data show that integrin engagement and the activation of integrin-mediated signaling pathways are essential in the induction of EHV-1-infected CD172a + adhesion to EC.”
Quote in the paperSubstantial claim
- Classified as probably not a substantial claim. 25
- May combine two claims. 26
- The quote only partly states this claim. 3
Probably not a substantial claim: may not pass S3 (load-bearing).
Premisecitation§ RESULTS · p. 4α4β1, αLβ2, and αVβ3 integrins are known to mediate firm monocyte adhesion to endothelium.
“These integrins are known to play a role in mediating firm adhesion of monocytes to the endothelium (24-26).”
Quote in the paperNot a substantial claim1 of 3 sources checked- cites [24] 24. Elangbam CS, Qualls CW, Dahlgren RR. 1997. Cell adhesion moleculesupdate. Vet Pathol 34: 61-73. http://dx.doi.org/10.1177/0300985897034 00113. source
- cites [25] 25. Mazzone A, Ricevuti G. 1995. Leukocyte CD11/CD18 integrins: biological and clinical relevance. Haematologica 80: 161-175.
Couldn’t be matched to a paper in Crossref, so it can’t be checked.
- cites [26] 26. Weerasinghe D, McHugh KP, Ross FP, Brown EJ, Gisler RH, Imhof BA. 1998. A role for the alphavbeta3 integrin in the transmigration of monocytes. J Cell Biol 142: 595-607. http://dx.doi.org/10.1083/jcb.142.2.595. source
The Discussion (page not available in our copy) says α4β1 promotes initial adhesion and αLβ2 mediates tight adhesion to endothelium. It instead describes αvβ3 as reducing αLβ2-dependent adhesion to ICAM-1 to enable monocyte migration, not as mediating firm adhesion. (LLM, conf=0.89)
Experimental — accuracy not yet measured
Classified as probably not a substantial claim.
Premisecitation§ RESULTS · p. 5Herpesvirus binding to integrins activates PI3K/Akt and downstream ERK/MAPK.
“Binding of herpesviruses to integrins is known to activate cellular pathways, including PI3K/Akt (PI3K stands for phosphatidylinositol 3-kinase and Akt for protein kinase B), one of the early events of integrinligand interaction, and the downstream ERK/MAPK (MAPK stands for mitogen-activated protein kinase and ERK for extracellular signal-regulated kinases).”
Quote in the paperSubstantial claim1 of 3 sources checked- cites [15] 15. Smith MS, Bivins-Smith ER, Tilley AM, Bentz GL, Chan G, Minard J, Yurochko AD. 2007. Roles of phosphatidylinositol 3-kinase and NFkappaB in human cytomegalovirus-mediated monocyte diapedesis and adhesion: strategy for viral persistence. J Virol 81: 7683-7694. http://dx.doi .org/10.1128/JVI.02839-06. source
- cites [27] 27. Chang MY, Huang DY, Ho FM, Huang KC, Lin WW. 2012. PKCdependenthumanmonocyteadhesionrequiresAMPKandSykactivation. PLoS One 7: e40999. http://dx.doi.org/10.1371/journal.pone.0040999. source
The Discussion and the section on PKC-dependent ERK activation concern PMA-induced monocyte adhesion, integrins, and ERK signaling. Searches for herpesvirus binding and PI3K/Akt found no passage addressing whether viral binding to integrins activates PI3K/Akt or downstream ERK/MAPK. (LLM, conf=0.78)
Experimental — accuracy not yet measured - cites [28] 28. Naranatt PP, Akula SM, Zien CA, Krishnan HH, Chandran B. 2003. Kaposi's sarcoma-associated herpesvirus induces the phosphatidylinositol 3-kinase-PKC-zeta-MEK-ERK signaling pathway in target cells early during infection: implications for infectivity. J Virol 77: 1524-1539. http: //dx.doi.org/10.1128/JVI.77.2.1524-1539.2003. source
May combine two claims.
Premisecitation§ DISCUSSION · p. 10EHV-1 infection of endothelial cells also induced extravasation of mononuclear cells, resulting in perivascular cuffing and local hemorrhage.
“EHV-1 infection of endothelial cells also induced extravasation of mononuclear cells, resulting in perivascular cuffing and local hemorrhage (50).”
Quote in the paperSubstantial claim1 of 1 sources checked- cites [50] 50. Stierstorfer B, Eichhorn W, Schmahl W, Brandmüller C, Kaaden OR, Neubauer A. 2002. Equine herpesvirus type 1 (EHV-1) myeloencephalopathy: a case report. J Vet Med B Infect Dis Vet Public Health 49: 37-41. http://dx.doi.org/10.1046/j.1439-0450.2002.00537.x. source
The Pathohistology section reports perivascular mononuclear cuffing and hemorrhagic areas, and suggests that vascular endothelium is an initial site of EHV-1 replication in the CNS. It does not establish that endothelial infection induced mononuclear-cell extravasation or caused the cuffing and hemorrhage; the claim states that causal sequence more definitively than the paper does. (LLM, conf=0.86)
Experimental — accuracy not yet measured
Probably not a substantial claim: may not pass S3 (load-bearing).
Premisecitation§ DISCUSSION · p. 9Several herpesviruses, such as PRV, HSV-1, HHV-8, and HCMV, activate integrin-mediated ERK-MAPK and PI3K signaling pathways through interactions of viral glycoproteins with putative receptors on host cell plasma membranes.
“Several herpesviruses, such as pseudorabies virus (PRV), herpes simplex virus 1 (HSV-1), human herpesvirus 8 (HHV-8), and HCMV, have been reported to activate integrin-mediated ERK-MAPK and PI3K signaling pathways through the interactions of viral glycoproteins with putative receptors present at the plasma membranes of host cells (23, 28, 33-35).”
Quote in the paperNot a substantial claim3 of 5 sources checked- cites [23] 23. Setas Pontes M, Devriendt B, Favoreel HW. 2015. Pseudorabies virus triggers glycoprotein gE-mediated ERK1/2 activation and ERK1/2dependent migratory behavior in T cells. J Virol 89: 2149-2156. http://dx .doi.org/10.1128/JVI.02549-14. source
The Discussion (p. 6) reports that PRV glycoprotein gE triggers ERK1/2 activation in T cells. It describes binding to an unidentified cell-surface receptor as speculative, and does not establish integrin-mediated signaling or PI3K activation. The same discussion does not establish the claimed mechanism for HSV-1, HHV-8, or HCMV. (LLM, conf=0.87)
Experimental — accuracy not yet measured - cites [28] 28. Naranatt PP, Akula SM, Zien CA, Krishnan HH, Chandran B. 2003. Kaposi's sarcoma-associated herpesvirus induces the phosphatidylinositol 3-kinase-PKC-zeta-MEK-ERK signaling pathway in target cells early during infection: implications for infectivity. J Virol 77: 1524-1539. http: //dx.doi.org/10.1128/JVI.77.2.1524-1539.2003. source
- cites [33] 33. Zheng K, Xiang Y, Wang X, Wang Q, Zhong M, Wang S, Wang X, Fan J, Kitazato K, Wanga Y. 2014. Epidermal growth factor receptor-PI3K signaling controls cofilin activity to facilitate herpes simplex virus 1 entry into neuronal cells. mBio 5: e00958-13. http://dx.doi.org/10.1128/mBio .00958-13. source
- cites [34] 34. Reeves MB, Breidenstein A, Compton T. 2012. Human cytomegalovirus activation of ERK and myeloid cell leukemia-1 protein correlates with survival of latently infected cells. Proc Natl Acad Sci U S A 109: 588-593. http://dx.doi.org/10.1073/pnas.1114966108. source
The Discussion (page not provided) describes HCMV gB interactions with cellular integrins during entry and reports HCMV activation of PI3K; the Results link HCMV entry to ERK–MAPK activation. The paper does not establish that integrins mediate both signaling pathways, and its passages do not address PRV, HSV-1, or HHV-8. (LLM, conf=0.84)
Experimental — accuracy not yet measured - cites [35] 35. Nogalski MT, Chan G, Stevenson EV, Gray S, Yurochko AD. 2011. Human cytomegalovirus-regulated paxillin in monocytes links cellular pathogenic motility to the process of viral entry. J Virol 85: 1360-1369. http://dx.doi.org/10.1128/JVI.02090-10. source
Probably not a substantial claim: may not pass S3 (load-bearing).
Premisecitation§ DISCUSSION · p. 10Production of TNFα induces cellular changes in monocytic cells, resulting in increased viral gene expression.
“This is in agreement with previous studies on HCMV and HIV infections, which demonstrated that the production of TNF𝛂 induces cellular changes in monocytic cells, resulting in increased viral gene expression (41, 42).”
Quote in the paperNot a substantial claim1 of 2 sources checked- cites [41] 41. Söderberg-Nauclér C, Kenneth NF, Nelson JA. 1997. Interferon-gamma and tumor necrosis factor-alpha specifically induce formation of cytomegalovirus-permissive monocyte-derived macrophages that are refractory to the antiviral activity of these cytokines. J Clin Invest 100: 31543163. http://dx.doi.org/10.1172/JCI119871. source
- cites [42] 42. Poli G, Kinter A, Justement JS, Kehrl JH, Bresseler P, Stanley S, Fauci A. 1990. Tumor necrosis factor-alpha functions in an autocrine manner in the induction of human immunodeficiency virus expression. Proc Natl Acad Sci U S A 87: 782-785. http://dx.doi.org/10.1073/pnas.87.2.782. source
Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
Premisecitation§ DISCUSSION · p. 9L-selectins are expressed on myeloid cells and play an important role in the initial interaction or tethering of monocytes to the endothelium.
“In addition to integrins, L-selectins are expressed on myeloid cells and have been shown to play an important role in the initial interaction with or tethering of monocytes to the endothelium (31, 32).”
Quote in the paperNot a substantial claim1 of 2 sources checked- cites [31] 31. Vestweber D, Blanks JE. 1999. Mechanisms that regulate the function of the selectins and their ligands. Physiol Rev 79: 181-213. source
- cites [32] 32. Giuffrè L, Cordey A, Monai N, Tardy T, Schapira M, Spertini O. 1997. Monocyte adhesion to activated aortic endothelium: role of L-selectin and heparan sulfate proteoglycans. J Cell Biol 136: 945-956. http://dx.doi.org /10.1083/jcb.136.4.945. source
Classified as probably not a substantial claim. May combine two claims.
Premisecitation§ RESULTS · p. 5PI3K/Akt and ERK/MAPK pathways have been linked to adhesion control and to herpesvirus infection and replication in target cells.
“Both these pathways have been implicated in the manipulation of cellular functions, such as adhesion, and in the control of herpesvirus infection and replication in target cells (15, 27, 28).”
Quote in the paperSubstantial claim1 of 3 sources checked- cites [15] 15. Smith MS, Bivins-Smith ER, Tilley AM, Bentz GL, Chan G, Minard J, Yurochko AD. 2007. Roles of phosphatidylinositol 3-kinase and NFkappaB in human cytomegalovirus-mediated monocyte diapedesis and adhesion: strategy for viral persistence. J Virol 81: 7683-7694. http://dx.doi .org/10.1128/JVI.02839-06. source
- cites [27] 27. Chang MY, Huang DY, Ho FM, Huang KC, Lin WW. 2012. PKCdependenthumanmonocyteadhesionrequiresAMPKandSykactivation. PLoS One 7: e40999. http://dx.doi.org/10.1371/journal.pone.0040999. source
The Results section, “PKC-dependent ERK activation contributes to monocyte adhesion” (page not provided), links ERK signaling to monocyte adhesion. The copy does not link PI3K/Akt to adhesion or address herpesvirus infection or replication; its use of viral infection concerns an adenovirus experimental vector. (LLM, conf=0.88)
Experimental — accuracy not yet measured - cites [28] 28. Naranatt PP, Akula SM, Zien CA, Krishnan HH, Chandran B. 2003. Kaposi's sarcoma-associated herpesvirus induces the phosphatidylinositol 3-kinase-PKC-zeta-MEK-ERK signaling pathway in target cells early during infection: implications for infectivity. J Virol 77: 1524-1539. http: //dx.doi.org/10.1128/JVI.77.2.1524-1539.2003. source
Classified as probably not a substantial claim. May combine two claims.
Premisecitation§ DISCUSSION · p. 9EHV-1 envelope glycoprotein D (gD) interacts with αV integrins present on PBMC and is critical for viral entry via endocytosis.
“To date, the EHV-1 envelope glycoprotein D (gD) has been shown to interact with 𝛂 V integrins present on PBMC and to be critical for viral entry via endocytosis (36).”
Quote in the paperSubstantial claim1 of 1 sources checked- cites [36] 36. Van de Walle G, Peters ST, VanderVen BC, O'Callaghan DJ, Osterrieder N. 2008. Equine herpesvirus 1 entry via endocytosis is facilitated by alphaV integrins and an RSD motif in glycoprotein D. J Virol 82: 1185911868. http://dx.doi.org/10.1128/JVI.00868-08. source
Probably not a substantial claim: may not pass S3 (load-bearing).
Premisecitation§ DISCUSSION · p. 9Contacts with EC enhanced HIV-1 replication in infected monocytes and macrophages.
“These findings corroborate in vitro studies on HIV, demonstrating that contacts with EC enhanced HIV-1 replication in infected monocytes and macrophages (39, 40).”
Quote in the paperNot a substantial claim1 of 2 sources checked- cites [39] 39. Fan ST, Hsia K, Edington TS. 1994. Upregulation of human immunodeficiency virus-1 in chronically infected monocytic cell line by both contact with endothelial cells and cytokines. Blood 84: 1565-1572. source
The ‘Induction of HIV-I expression by endothelial cell’ section reports increased HIV-1 p24 production when infected U1 cells were cocultured with endothelial monolayers. The paper identifies U1 as a promonocytic cell line; it does not establish the same result in infected macrophages. (LLM, conf=0.86)
Experimental — accuracy not yet measured - cites [40] 40. Gilles PN, Lathey JL, Spector SA. 1995. Replication of macrophagetropic and T-cell-tropic strains of human immunodeficiency virus type 1 - is augmented by macrophage-endothelial cell contact. J Virol 69: 21332139. source
May combine two claims.
Findingown data§ IMPORTANCE · p. 1Both direct cell-to-cell contact and soluble factors secreted by endothelial cells activate EHV-1 replication in CD172a+ cells.
“We found that both cell-to-cell contacts and the secretion of soluble factors by EC activate EHV-1 replication in CD172a ÷ cells.”
Quote in the paperSubstantial claimThe quote only partly states this claim.
Findingown data§ RESULTS · p. 5Antibody blockade of α4β1, αLβ2, and αVβ3 integrins lowers adhesion of EHV-1-inoculated CD172a+ cells to endothelial cells about threefold after two hours relative to isotype controls.
“Pretreatment of EHV-1-inoculated CD172a + cells with blocking antibodies resulted in a 3-fold decrease in adhesion to EC after 2 h of coculture compared to isotype controls ( P < 0.001) (Fig. 2B andC).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown data§ ABSTRACT · p. 1EHV-1 replication is higher in adherent CD172a+ cells and tracks with TNFα production.
“EHV-1 replication was enhanced in adherent CD172a ÷ cells, which correlates with the production of tumor necrosis factor alpha (TNF𝛂 ).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown data§ RESULTS · p. 7For EHV-1 strain 97P70, the percentage of IEP-positive CD172a+ cells remained stable at 4% from 2 to 6 h postadhesion and increased significantly to 10% at 12 h postadhesion.
“In contrast, the percentage of IEP-positive CD172a + cells remained stable from 2 to 6 h p.a. (4%) and significantly increased to 10% at 12 h p.a. for EHV-1 strain 97P70 ( P < 0.01) (Fig. 4A, right).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown data§ RESULTS · p. 7At 12 h postadhesion, gB expression was significantly increased in IEP-positive cells inoculated with EHV-1 strain 97P70 and cocultured with EC monolayers compared to cells cultured on plastic (P<0.05), while an upward trend was observed for strain 03P37.
“An upward trend and a significant increase in gB expression ( P < 0.05) were observed in IEP-positive cells inoculated with EHV-1 strains 03P37 and 97P70, respectively, and cocultured for 12 h with EC monolayers compared to cells cultured on plastic (Fig. 4C).”
Quote in the paperSubstantial claimMay combine two claims.
Findingown data§ RESULTS · p. 8gB protein was expressed only in the cytoplasm of infected CD172a+ cells adherent to EC, and no gB expression was detected at the plasma membrane after 12 h coculture with EC.
“The gB protein was expressed only in the cytoplasm of these cells, and no gB expression was detected at the plasma membrane in CD172a + cells after 12 h coculture with EC.”
Quote in the paperSubstantial claimMay combine two claims.
Findingown data§ DISCUSSION · p. 9The percentage of IEP-positive CD172a+ cells significantly increased from 2 to 12 h p.a. for EHV-1 strain 03P37 while it increased only at 12 h p.a. for EHV-1 strain 97P70.
“Interestingly, we demonstrated that the percentage of IEP-positive CD172a + cells significantly increased from 2 to 12 h p.a. for EHV-1 strain 03P37 while it increased only at 12 h p.a. for EHV-1 (97P70).”
Quote in the paperSubstantial claimThe quote only partly states this claim.
Findingown data§ RESULTS · p. 8The number of gB-positive cells among IEP-positive CD172a+ cells was significantly higher (P<0.001) when stimulated with 10 ng/ml equine recombinant TNFα at 12 h p.i.
“We found that the number of cells positive for gB among IEP-positive cells was significantly higher ( P < 0.001) when stimulated with equine recombinant TNF𝛂 (Fig. 5B).”
Quote in the paperSubstantial claimMay combine two claims.
Interpretationown data§ ABSTRACT · p. 1EHV-1 infection causes CD172a+ cells to adhere to endothelial cells, and this adhesion boosts viral replication.
“Our results demonstrated that EHV-1 infection induces adhesion of CD172a ÷ cells to EC, which enhances viral replication, but that transfer of viral material from CD172a ÷ cells to EC is a very specific and rare event.”
Quote in the paperSubstantial claimMay combine two claims.
Premisecitation§ IMPORTANCE · p. 2Infection of endothelial cells in the late-gravid uterus or CNS vasculature occurs through cell-to-cell contact between infected PBMC and endothelial cells and can happen despite virus-neutralizing antibodies.
“Studies have shown that the infection of EC located in the vasculature of the late-gravid uterus or CNS was mediated by cellto-cell contacts between infected PBMC and EC and occurred even in the presence of virus-neutralizing antibodies (18, 19).”
Quote in the paperSubstantial claim0 of 2 sources checked- cites [18] 18. Smith DJ, Hamblin AS, Edington N. 2001. Infection of endothelial cells with equine herpesvirus-1 (EHV-1) occurs where there is activation of putative adhesion molecules: a mechanism for transfer of virus. Equine Vet J 33: 138-142. source
- cites [19] 19. Goehring LS, Hussey GS, Ashton LV, Schenkel AR, Lunn DP. 2011. Infection of central nervous system endothelial cells by cell-associated EHV-1. Vet Microbiol 148: 389-395. http://dx.doi.org/10.1016/j.vetmic .2010.08.030. source
Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
Premisecitation§ IMPORTANCE · p. 1EHV-1 first replicates in upper respiratory tract epithelium and then spreads through the body via cell-associated viremia in PBMC to organs such as the pregnant uterus or CNS.
“The virus first replicates in the epithelial cells of the upper respiratory tract and disseminates through the body via a cell-associated viremia in peripheral blood mononuclear cells (PBMC) to target organs, such as the pregnant uterus or central nervous system (CNS).”
Quote in the paperNot a substantial claim0 of 3 sources checked- cites [5] 5. Edington N, Smith B, Griffiths L. 1991. The role of endothelial cell infection in the endometrium, placenta and fetus of equid herpesvirus 1 (EHV-1) abortions. J Comp Pathol 104: 379-387. http://dx.doi.org/10 .1016/S0021-9975(08)80148-X. source
- cites [6] 6. Smith KC, Mumford JA, Lakhani K. 1996. A comparison of equid herpesvirus-1 (EHV-1) vascular lesions in the early versus late pregnant equine uterus. J Comp Pathol 114: 231-247. http://dx.doi.org/10.1016 /S0021-9975(96)80045-4. source
- cites [7] 7. Wilson WD. 1997. Equine herpesvirus 1 myeloencephalopathy. Vet Clin North Am Equine Pract 13: 53-72. source
Classified as probably not a substantial claim. May combine two claims.
Premisecitation§ IMPORTANCE · p. 2Activation of endothelial adhesion molecules may participate in virus transfer from infected PBMC to endothelial cells and may shape EHV-1 tissue restriction.
“In addition, Smith et al. (18) provided evidence that activation of EC adhesion molecules may be involved in the transfer of virus from infected PBMC to EC and may determine the restricted tissue specificity of EHV-1.”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [18] 18. Smith DJ, Hamblin AS, Edington N. 2001. Infection of endothelial cells with equine herpesvirus-1 (EHV-1) occurs where there is activation of putative adhesion molecules: a mechanism for transfer of virus. Equine Vet J 33: 138-142. source
May combine two claims.
Premisecitation§ IMPORTANCE · p. 1CD172a+ cells act as a Trojan horse that helps EHV-1 reach target organs and avoid immune surveillance.
“We previously reported that CD172a + cells serve as a 'Trojan horse' to facilitate the spread of EHV-1 to target organs and to evade immunosurveillance (10).”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [10] 10. Laval K, Favoreel HW, Nauwynck HJ. 2015. Equine herpesvirus type 1 (EHV-1) replication is delayed in CD172a + monocytic cells and con- source
May combine two claims.
Interpretationown data§ ABSTRACT · p. 1Transfer of viral material from CD172a+ cells to endothelial cells is a rare and highly specific event.
“Our results demonstrated that EHV-1 infection induces adhesion of CD172a ÷ cells to EC, which enhances viral replication, but that transfer of viral material from CD172a ÷ cells to EC is a very specific and rare event.”
Quote in the paperSubstantial claimClassified as probably not a substantial claim.
Premisecitation§ IMPORTANCE · p. 1Endothelial cell-monocyte interactions are central to herpesvirus pathogenesis.
“EC are actively involved in a wide variety of pathological processes, such as thrombosis and vasculitis, and endothelial cellmonocyte interactions are known to play a central role in the pathogenesis of herpesviruses infections.”
Quote in the paperSubstantial claimMay combine two claims.
Interpretationown data§ RESULTS · p. 4EHV-1 gene expression is not needed for the virus to trigger CD172a+ cell adhesion to endothelial cells; entry alone is enough.
“These data indicate that active EHV-1 gene expression was not required to induce adhesion of CD172a + cells to EC monolayers, and thus, viral entry was sufficient to activate CD172a + cell adhesion to EC.”
Quote in the paperSubstantial claimClassified as probably not a substantial claim.
Premisenone given§ IMPORTANCE · p. 2The exact mechanism by which EHV-1 moves from monocytic cells to endothelial cells remains unknown.
“However, the precise mechanism underlying the transmission of EHV-1 from monocytic cells to EC is still unclear.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Premisecitation§ IMPORTANCE · p. 1HCMV infection of endothelial cells promotes adhesion and migration of naïve monocytes through the endothelium.
“For instance, human cytomegalovirus (HCMV) infection of EC has been shown to promote naïve-monocyte adhesion to and migration through the endothelium”
Quote in the paperNot a substantial claim0 of 2 sources checked- cites [14] 14. Bentz GL, Jarquin-Pardo M, Chan G, Smith MS, Sinzger C, Yurochko AD. 2006. Human cytomegalovirus (HCMV) infection of endothelial cells promotes naive monocyte extravasation and transfer of productive virus to enhance hematogenous dissemination of HCMV. J Virol 80: 11539-11555. http://dx.doi.org/10.1128/JVI.01016-06. source
- cites [15] 15. Smith MS, Bivins-Smith ER, Tilley AM, Bentz GL, Chan G, Minard J, Yurochko AD. 2007. Roles of phosphatidylinositol 3-kinase and NFkappaB in human cytomegalovirus-mediated monocyte diapedesis and adhesion: strategy for viral persistence. J Virol 81: 7683-7694. http://dx.doi .org/10.1128/JVI.02839-06. source
Probably not a substantial claim: may not pass S3 (load-bearing). The quote only partly states this claim.
Study design and conductown data§ RESULTS · p. 5Technical limitations prevented testing whether EHV-1 infection increases ERK/MAPK and PI3K activity in CD172a+ cells.
“Thus, we could not test whether EHV-1 infection upregulates ERK/MAPK and PI3K activities in CD172a + cells.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S4 (based). May combine two claims.
Study design and conductnone given§ MATERIALS AND METHODS · p. 2The inhibitor concentrations and DMSO diluent used did not reduce cell viability.
“The concentration of inhibitors used in this study and the DMSO-based diluent did not decrease cell viability.”
Quote in the paperNot a substantial claimMay combine two claims.
Interpretationown data§ DISCUSSION · p. 9Adhesion was not completely inhibited by the addition of blocking antibodies, suggesting that other adhesion molecules may be involved in this process.
“However, adhesion was not completely inhibited by the addition of blocking antibodies, suggesting that other adhesion molecules may be involved in this process.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductnone given§ DISCUSSION · p. 9There is no antibody available against L-selectin that cross-reacts specifically with equine monocytes to allow further experiments.
“Unfortunately, there is no antibody available against L-selectin that cross-reacts specifically with equine monocytes to allow further experiments.”
Quote in the paperNot a substantial claimMay combine two claims.
Interpretationown data§ DISCUSSION · p. 10TNFα was produced via direct interaction between CD172a+ cells and EC, and TNFα triggered the enhanced EHV-1 replication observed in CD172a+ cells.
“In addition, we showed that TNF𝛂 was produced via direct interaction between CD172a + cells and EC and that TNF𝛂 triggered the enhanced EHV-1 replication observed in CD172a + cells.”
Quote in the paperSubstantial claimClassified as probably not a substantial claim.
Premisecitation§ DISCUSSION · p. 10EHV-1 infection of endothelial cells of the pregnant uterus or CNS results in damage to the microvasculature of target organs due to initiation of an inflammatory cascade, vasculitis, and microthrombosis.
“Previous in vivo studies showed that EHV-1 infection of endothelial cells of the pregnant uterus or CNS resulted in damage to the microvasculature of target organs due to the initiation of an inflammatory cascade, vasculitis, and microthrombosis (47-49).”
Quote in the paperSubstantial claim0 of 3 sources checked- cites [47] 47. Edington N, Bridges CG, Patel JR. 1986. Endothelial cell infection and thrombosis in paralysis caused by equid herpesvirus-1: equine stroke. Arch Virol 90: 111-124. http://dx.doi.org/10.1007/BF01314149. source
- cites [48] 48. Patel JR, Edington N, Mumford JA. 1982. Variation in cellular tropism between isolates of equine herpesvirus-1 in foals. Arch Virol 74: 41-51. http://dx.doi.org/10.1007/BF01320781. source
- cites [49] 49. Smith KC, Whitwell KE, Binns MM, Dolby CA, Hannant D, Mumford JA. 1992. Abortion of virologically negative foetuses following experimental challenge of pregnant pony mares with equid herpesvirus 1. Equine Vet J 24: 256-259. http://dx.doi.org/10.1111/j.2042-3306.1992.tb02830.x. source
Probably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductown data§ DISCUSSION · p. 9It was technically impossible to assess any potential upregulation of integrin expression on EHV-1-inoculated cells compared to mock-inoculated cells due to a low percentage of infected CD172a+ cells detected.
“Indeed, the antibodies used in this study do not discriminate between inactive and activated forms of the integrins, and it was technically impossible to assess any potential upregulation of integrin expression on EHV-1-inoculated cells compared to mock-inoculated cells, due to a low percentage of infected CD172a + cells detected.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
Study design and conductown data§ MATERIALS AND METHODS · p. 2Virus inactivation used short-wave UV light for 10 minutes, and absence of infectivity was confirmed by titration on RK-13 cells.
“For virus inactivation, a thin layer of viral suspension was exposed to short-wave UV light for 10 min. Absence of viral infectivity was checked by virus titration on RK-13 cells.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductown data§ MATERIALS AND METHODS · p. 4Experiments were run three times.
“Experiments were performed three times.”
Quote in the paperNot a substantial claimMay combine two claims.
Interpretationown data§ DISCUSSION · p. 10The enhanced EHV-1 replication observed in CD172a+ cells is a complex process involving both cell contacts and the production of soluble factors, with major implications for EHV-1 spread to EC.
“Taken together, our results indicate that the enhanced EHV-1 replication observed in CD172a + cells is a complex process involving both cell contacts and the production of soluble factors, with major implications for EHV-1 spread to EC (Fig. 7C).”
Quote in the paperSubstantial claimMay combine two claims.
Interpretationown data§ RESULTS · p. 7Adhesion of EHV-1-inoculated CD172a+ cells to EC speeds up viral replication in cells inoculated with strain 97P70, but to a lesser extent in cells inoculated with strain 03P37, compared to cells adherent to plastic.
“Taken together, these results show that the adhesion of EHV1-inoculated CD172a + cells to EC speeds up the viral replication process in CD172a + cells inoculated with EHV-1 strain 97P70, but to a lesser extent in CD172a + cells inoculated with EHV-1 strain 03P37, compared to EHV-1-inoculated CD172a + cells adherent to plastic.”
Quote in the paperSubstantial claimMay combine two claims.
Interpretationreasoning§ DISCUSSION · p. 11The interaction between EHV-1-infected CD172a+ cells and EC enhances EHV-1 replication in CD172a+ cells through cell-to-cell contacts and secretion of soluble factors, which subsequently facilitates viral transfer to EC.
“The interaction between EHV-1-infected CD172a + cells and EC enhances EHV-1 replication in CD172a + cells through cell-to-cell contacts and secretion of soluble factors, which subsequently facilitates viral transfer to EC.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
Study design and conductown data§ RESULTS · p. 7Three independent experiments were performed for the expression kinetics data, and data are represented as means plus SD.
“Three independent experiments were performed, and the data are represented as means plus SD.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductown data§ RESULTS · p. 7A two-way ANOVA test was performed to evaluate significant differences from the control.
“A two-way ANOVA test was performed to evaluate significant differences from the control (*, P < 0.05; **, P < 0.01).”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
Study design and conductnone given§ MATERIALS AND METHODS · p. 3Mock-inoculated negative controls and isotype-matched controls were included in staining.
“As a negative control, mock-inoculated cells were stained following the above-mentioned protocols. In addition, appropriate isotype-matched controls were included.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
Premisecitation§ DISCUSSION · p. 10TNFα secreted by EC is associated with increased HIV expression in monocytic cells.
“TNF𝛂 secreted by EC was also found to be associated with increased HIV expression in monocytic cells (43).”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [43] 43. Borghi MO, Panzeri P, Shattock R, Sozzani S, Dobrina A, Meroni PL. 2000. Interaction between chronically HIV-infected promonocytic cells and human umbilical vein endothelial cells: role of proinflammatory cytokines and chemokines in viral expression modulation. Clin Exp Immunol 120: 93-100. http://dx.doi.org/10.1046/j.1365-2249.2000.01186.x. source
Probably not a substantial claim: may not pass S3 (load-bearing).
Premisecitation§ DISCUSSION · p. 10Direct transfer of HCMV cytoplasmic material from infected to neighboring uninfected cells occurs.
“Our data are in line with the findings of Digel et al. (44), showing direct transfer of HCMV cytoplasmic material from infected to neighboring uninfected cells.”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [44] 44. Digel M, Sampaio KL, Jahn G, Sinzger C. 2006. Evidence for direct transfer of cytoplasmic material from infected to uninfected cells during cell-associated spread of human cytomegalovirus. J Clin Virol 37: 10-20. http://dx.doi.org/10.1016/j.jcv.2006.05.007. source
The paper relies on this without support.
The in vitro coculture model with immortalized endothelial cells reflects the relevant CD172a+ cell-endothelial cell interactions that occur in vivo.
“As the kinetics of CD172a + cell adhesion were similar in primary EC cultures and immortalized EC lines, the subsequent experiments were performed using immortalized EC only.”
“These findings give new insights into the complex pathogenesis of EHV-1.”
The authors state this as an assumption.
Monocytic cells play a key role in EHV-1 spread from the blood to target organs.
“Monocytic cells are assumed to play a key role in EHV-1 spread from the blood to target organs, but the precise interactions between monocytic cells and EC have not been unraveled.”
The paper relies on this without support.
The precise interactions between monocytic cells and EC have not been unraveled.
“Monocytic cells are assumed to play a key role in EHV-1 spread from the blood to target organs, but the precise interactions between monocytic cells and EC have not been unraveled.”
The paper relies on this without support.
TNFα is a potent inducer of viral gene expression in monocytes/macrophages.
“Since TNF𝛂 has been described as a potent inducer of viral gene expression in monocytes/macrophages, we investigated whether TNF𝛂 is produced during interaction of EHV-1-inoculated CD172a + cells and EC and whether TNF𝛂 enhances EHV-1 replication in CD172a + cells.”
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
No EHV-1-productive infection in PBMC was observed, although this does not exclude restricted productive virus replication at low levels.
“Most remarkably, no EHV-1-productive infection in PBMC was observed, which, however, does not exclude unambiguously restricted productive virus replication albeit at low levels (Drebert et al., 2015; Laval et al., 2015; Spiesschaert et al., 2015a).”
From Equid Herpesvirus-1 Exploits the Extracellular Matrix of Mononuclear Cells to Ensure Transport to Target Cells (2020) · cites this paper as Laval et al., 2015
The Results (p. 8) report that a small fraction of EHV-1-inoculated CD172a+ monocytic cells produced and transmitted infectious virus to endothelial cells: 0.1% and 0.05% for the two strains. These cells were isolated from PBMC (Materials and Methods, p. 2), so the paper reports rare productive infection in a PBMC-derived population rather than no observed productive infection. It does not establish the rate for PBMC as a whole. (LLM, conf=0.84)
Checked by a reasoning model — may be wrong.
Experimental — accuracy not yet measuredWhen infected PBMC reach the endothelium, the virus finds its way outside the PBMC to infect new cells, probably through cell-cell fusion.
“However, when infected PBMC reach the endothelium, the virus finds its way outside the PBMC to infect new cells, probably through cell-cell fusion (Laval et al., 2015; Spiesschaert et al., 2015a).”
From Equid Herpesvirus-1 Exploits the Extracellular Matrix of Mononuclear Cells to Ensure Transport to Target Cells (2020) · cites this paper as Laval et al., 2015
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
Each contradiction between two passages listed here was confirmed by an independent check.
Abstract transmission rate conflicts with reported assay rates
MinorThe abstract gives a transmission rate of 0.01% of infected cells, whereas the Results report 0.1% and 0.05% of all inoculated CD172a+ cells. An infected-cell subset cannot have a lower transmission percentage than the entire inoculated population if these statements describe the same outcome.
“0.01% of infected CD172a ÷ cells transmitted infectious virus to neighboring cells.”
“After 5 days of coculture of inoculated CD172a + cells with EC monolayers, we showed that 0.1% and 0.05% of CD172a + cells inoculated with EHV-1 strains 03P37 and 97P70, respectively, transmitted the infection to EC.”
Passage count for strain 03P37 does not total six
MinorThe stated four passages in EEL cells plus one in RK-13 cells total five, not the six passages reported for the virus stocks.
Reported 6 · recomputed 5
“The virus stocks used for inoculation were at the sixth passage, with five passages in equine embryonic lung cells (EEL) and one passage in RK-13 cells for strain 97P70 and four passages in EEL and one passage in RK-13 cells for strain 03P37.”
Cites
Infectious diseases of livestock
Coetzer, J. A. W., & Tustin, R. C. (2004) · Oxford University Press eBooks · Cited by 1,226 · see citing works
Mechanisms That Regulate the Function of the Selectins and Their Ligands
VESTWEBER, D., & BLANKS, J. E. (1999) · Physiological Reviews · doi:10.1152/physrev.1999.79.1.181 · Cited by 932 · see citing works
(1986) · PubMed · Cited by 467 · see citing works
Poli, G., Kinter, A., Justement, J. S., Kehrl, J. H., Bressler, P., Stanley, S., & Fauci, A. S. (1990) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.87.2.782 · Cited by 457 · see citing works
Luscinskas, F. W., Kansas, G. S., Ding, H. T., Pizcueta, P. M., Schleiffenbaum, B. E., Tedder, T. F., & Gimbrone, M. A. (1994) · The Journal of Cell Biology · doi:10.1083/jcb.125.6.1417 · Cited by 376 · see citing works
Carlos, T. M., Kovach, N. L., Schwartz, B. R., Rosa, M., Newman, B., Wayner, E. A., Benjamin, C. D., Osborn, L., Lobb, R. R., & Harlan, J. (1991) · Blood · doi:10.1182/blood.v77.10.2266.2266 · Cited by 318 · see citing works
Huang, S., Endo, R. I., & Nemerow, G. R. (1995) · Journal of Virology · doi:10.1128/jvi.69.4.2257-2263.1995 · Cited by 292 · see citing works
Patel, J., & Heldens, J. G. M. (2004) · The Veterinary Journal · doi:10.1016/j.tvjl.2004.04.018 · Cited by 287 · see citing works
Cell Adhesion Molecules—Update
Elangbam, C. S., Qualls, C. W., & Dahlgren, R. R. (1997) · Veterinary Pathology · doi:10.1177/030098589703400113 · Cited by 215 · see citing works
(1995) · PubMed · Cited by 215 · see citing works
Endothelial cell infection and thrombosis in paralysis caused by equid herpesvirus-1: equine stroke.
Edington, N., Bridges, C. G., & Patel, J. R. (1986) · Archives of Virology · doi:10.1007/bf01314149 · Cited by 213 · see citing works
Naranatt, P. P., Akula, S. M., Zien, C. A., Krishnan, H. H., & Chandran, B. (2003) · Journal of Virology · doi:10.1128/jvi.77.2.1524-1539.2003 · Cited by 175 · see citing works
Gerna, G., Percivalle, E., Baldanti, F., Sozzani, S., Lanzarini, P., Genini, E., Lilleri, D., & Revello, M. G. (2000) · Journal of Virology · doi:10.1128/jvi.74.12.5629-5638.2000 · Cited by 171 · see citing works
Söderberg-Nauclér, C., Fish, K. N., & Nelson, J. A. (1997) · Journal of Clinical Investigation · doi:10.1172/jci119871 · Cited by 159 · see citing works
The equine immune response to equine herpesvirus-1: The virus and its vaccines
Kydd, J. H., Townsend, H. G. G., & Hannant, D. (2006) · Veterinary Immunology and Immunopathology · doi:10.1016/j.vetimm.2006.01.005 · Cited by 134 · see citing works
Variation in cellular tropism between isolates of equine herpesvirus-1 in foals.
Patel, J. R., Edington, N., & Mumford, J. A. (1982) · Archives of Virology · doi:10.1007/bf01320781 · Cited by 132 · see citing works
A Role for the αvβ3 Integrin in the Transmigration of Monocytes
Weerasinghe, D., McHugh, K. P., Ross, F. P., Brown, E. J., Gisler, R. H., & Imhof, B. A. (1998) · The Journal of Cell Biology · doi:10.1083/jcb.142.2.595 · Cited by 130 · see citing works
Bentz, G. L., Jarquin-Pardo, M., Chan, G., Smith, M. S., Sinzger, C., & Yurochko, A. D. (2006) · Journal of Virology · doi:10.1128/jvi.01016-06 · Cited by 130 · see citing works
Edington, N., Smyth, B., & Griffiths, L. (1991) · Journal of Comparative Pathology · doi:10.1016/s0021-9975(08)80148-x · Cited by 123 · see citing works
Giuffrè, L., Cordey, A.-S., Monai, N., Tardy, Y., Schapira, M., & Spertini, O. (1997) · The Journal of Cell Biology · doi:10.1083/jcb.136.4.945 · Cited by 122 · see citing works
Zheng, K., Xiang, Y., Wang, X., Wang, Q., Zhong, M., Wang, S., Wang, X., Fan, J., Kitazato, K., & Wang, Y. (2014) · mBio · doi:10.1128/mbio.00958-13 · Cited by 118 · see citing works
Equine herpesvirus 1 myeloencephalopathy.
Wilson, W. D. (1997) · Veterinary Clinics of North America: Equine Practice · doi:10.1016/s0749-0739(17)30255-9 · Cited by 113 · see citing works
SMITH, K. C., WHITWELL, K. E., BINNS, M. M., DOLBY, C. A., HANNANT, D., & MUMFORD, J. A. (1992) · Equine Veterinary Journal · doi:10.1111/j.2042-3306.1992.tb02830.x · Cited by 102 · see citing works
Smith, M. S., Bentz, G. L., Smith, P. M., Bivins-Smith, E. R., & Yurochko, A. D. (2004) · Journal of Leukocyte Biology · doi:10.1189/jlb.1203621 · Cited by 86 · see citing works
Reeves, M. B., Breidenstein, A., & Compton, T. (2012) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1114966108 · Cited by 82 · see citing works
(1987) · PubMed · Cited by 82 · see citing works
Evaluation of orally administered valacyclovir in experimentally EHV1-infected ponies
Garré, B., Gryspeerdt, A. C., Croubels, S., Backer, J. F. A. D., & Nauwynck, H. (2008) · Veterinary Microbiology · doi:10.1016/j.vetmic.2008.09.062 · Cited by 73 · see citing works
Self-citationFOOTE, C. E., Love, D. N., Gilkerson, J. R., & Whalley, J. M. (2004) · Equine Veterinary Journal · doi:10.2746/0425164044890634 · Cited by 70 · see citing works
O'Neill, T., Kydd, J. H., Allen, G. P., Wattrang, E., Mumford, J. A., & Hannant, D. (1999) · Veterinary Immunology and Immunopathology · doi:10.1016/s0165-2427(99)00037-9 · Cited by 69 · see citing works
Smith, K. C., Mumford, J. A., & Lakhani, K. (1996) · Journal of Comparative Pathology · doi:10.1016/s0021-9975(96)80045-4 · Cited by 68 · see citing works
Smith, M. S., Bivins-Smith, E. R., Tilley, A. M., Bentz, G. L., Chan, G., Minard, J., & Yurochko, A. D. (2007) · Journal of Virology · doi:10.1128/jvi.02839-06 · Cited by 64 · see citing works
van der Meulen, K. M., Nauwynck, H. J., Buddaert, W., & Pensaert, M. B. (2000) · Microbiology · doi:10.1099/0022-1317-81-1-21 · Cited by 57 · see citing works
Self-citationNogalski, M. T., Chan, G., Stevenson, E. V., Gray, S., & Yurochko, A. D. (2011) · Journal of Virology · doi:10.1128/jvi.02090-10 · Cited by 56 · see citing works
PKC-Dependent Human Monocyte Adhesion Requires AMPK and Syk Activation
Chang, M.-Y., Huang, D.-Y., Ho, F.-M., Huang, K.-C., & Lin, W.-W. (2012) · PLoS ONE · doi:10.1371/journal.pone.0040999 · Cited by 54 · see citing works
Van de Walle, G. R., Peters, S. T., VanderVen, B. C., O'Callaghan, D. J., & Osterrieder, N. (2008) · Journal of Virology · doi:10.1128/jvi.00868-08 · Cited by 54 · see citing works
van der Meulen, K. M., Nauwynck, H., & Pensaert, M. B. (2003) · Journal of General Virology · doi:10.1099/vir.0.18864-0 · Cited by 43 · see citing works
Self-citationIn vitro characterisation of high and low virulence isolates of equine herpesvirus-1 and -4
Tearle, J. P., Smith, K. C., Platt, A. J., Hannant, D., Davis-Poynter, N. J., & Mumford, J. A. (2003) · Research in Veterinary Science · doi:10.1016/s0034-5288(03)00031-6 · Cited by 41 · see citing works
Equine Herpesvirus Type 1 (EHV‐1) Myeloencephalopathy: a Case Report
STIERSTORFER, B., EICHHORN, W., SCHMAHL, W., BRANDMÜLLER, C., KAADEN, O. ‐., & NEUBAUER, A. (2002) · Journal of Veterinary Medicine, Series B · doi:10.1046/j.1439-0450.2002.00537.x · Cited by 41 · see citing works
SMITH, D. J., HAMBLIN, A. S., & EDINGTON, N. (2001) · Equine Veterinary Journal · doi:10.1111/j.2042-3306.2001.tb00591.x · Cited by 39 · see citing works
Fan, S., Hsia, K., & Edgington, T. (1994) · Blood · doi:10.1182/blood.v84.5.1567.1567 · Cited by 38 · see citing works
Gilles, P. N., Lathey, J. L., & Spector, S. A. (1995) · Journal of Virology · doi:10.1128/jvi.69.4.2133-2139.1995 · Cited by 38 · see citing works
A local epidemic of equine herpesvirus 1-induced neurological disorders in belgium
van der Meulen, K., Vercauteren, G., Nauwynck, H., & Pensaert, M. B. (2003) · Vlaams Diergeneeskundig Tijdschrift · doi:10.21825/vdt.89249 · Cited by 38 · see citing works
Self-citationDigel, M., Sampaio, K. L., Jahn, G., & Sinzger, C. (2006) · Journal of Clinical Virology · doi:10.1016/j.jcv.2006.05.007 · Cited by 36 · see citing works
Infection of central nervous system endothelial cells by cell-associated EHV-1
Goehring, L. S., Hussey, G. S., Ashton, L. V., Schenkel, A. R., & Lunn, D. P. (2011) · Veterinary Microbiology · doi:10.1016/j.vetmic.2010.08.030 · Cited by 34 · see citing works
Laval, K., Favoreel, H. W., & Nauwynck, H. J. (2015) · Journal of General Virology · doi:10.1099/vir.0.067363-0 · Cited by 29 · see citing works
Self-citationCarlos, T. M., Kovach, N. L., Schwartz, B., et al. (1991) · Blood · doi:10.1182/blood.v77.10.2266.bloodjournal77102266 · Cited by 22 · see citing works
Borghi, M. O., Panzeri, P., Shattock, R., Sozzani, S., Dobrina, A., & Meroni, P. L. (2001) · Clinical and Experimental Immunology · doi:10.1046/j.1365-2249.2000.01186.x · Cited by 21 · see citing works
Gryspeerdt, A. C., Vandekerckhove, A. P., Baghi, H. B., Van de Walle, G. R., & Nauwynck, H. J. (2012) · The Veterinary Journal · doi:10.1016/j.tvjl.2012.01.022 · Cited by 18 · see citing works
Self-citationSetas Pontes, M., Devriendt, B., & Favoreel, H. W. (2015) · Journal of Virology · doi:10.1128/jvi.02549-14 · Cited by 15 · see citing works
Self-citationTransmission of pseudorabies virus from immune-masked blood monocytes to endothelial cells
Van de Walle, G. R., Favoreel, H. W., Nauwynck, H. J., Mettenleiter, T. C., & Pensaert, M. B. (2003) · Journal of General Virology · doi:10.1099/vir.0.18796-0 · Cited by 15 · see citing works
Self-citationChiam, R., Smid, L., Kydd, J. H., Smith, K. C., Platt, A., & Davis-Poynter, N. J. (2005) · Veterinary Microbiology · doi:10.1016/j.vetmic.2005.11.005 · Cited by 9 · see citing works
Barbero (2014) · doi:10.1016/j · Cited by 1 · see citing works
Fan, S., Hsia, K., & Edgington, T. (1994) · Blood · doi:10.1182/blood.v84.5.1567.bloodjournal8451567
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Cited by
Sorted by citations
Deficient humoral responses and disrupted B-cell immunity are associated with fatal SFTSV infection
Song, P., Zheng, N., Liu, Y., Tian, C., Wu, X., Ma, X., Chen, D., Zou, X., Wang, G., Wang, H., et al. (2018) · Nature Communications · doi:10.1038/s41467-018-05746-9 · Cited by 170 · see citing works
Ayala‐Nunez, V., Follain, G., Delalande, F., Hirschler, A., Partiot, E., Hale, G., Bollweg, B. C., Roels, J. R. E., Chazal, M., Bakoa, F., et al. (2019) · Nature Communications · doi:10.1038/s41467-019-12408-x · Cited by 110 · see citing works
Chase, C. C. L., Fulton, R. W., O’Toole, D., et al. (2017) · Veterinary Microbiology · doi:10.1016/j.vetmic.2017.03.016 · Cited by 94 · see citing works
Gale, A. L., Linardi, R. L., McClung, G., et al. (2019) · Frontiers in Veterinary Science · doi:10.3389/fvets.2019.00178 · Cited by 57 · see citing works
The Pathogenesis and Immune Evasive Mechanisms of Equine Herpesvirus Type 1
Laval, K., Poelaert, K. C. K., Van Cleemput, J., Zhao, J., Vandekerckhove, A. P., Gryspeerdt, A. C., Garré, B., van der Meulen, K., Baghi, H. B., Dubale, H. N., et al. (2021) · Frontiers in Microbiology · doi:10.3389/fmicb.2021.662686 · Cited by 54 · see citing works
Self-citationViral-Immune Cell Interactions at the Maternal-Fetal Interface in Human Pregnancy
Parker, E. L., Silverstein, R. B., Verma, S., et al. (2020) · Frontiers in Immunology · doi:10.3389/fimmu.2020.522047 · Cited by 51 · see citing works
Kochagul, V., Srivorakul, S., Boonsri, K., et al. (2018) · Scientific Reports · doi:10.1038/s41598-018-22968-5 · Cited by 41 · see citing works
How do oxyphytosterols affect human health?
Wang, M., & Lu, B. (2018) · Trends in Food Science & Technology · doi:10.1016/j.tifs.2018.07.002 · Cited by 36 · see citing works
Equine Herpesvirus 1 Bridles T Lymphocytes To Reach Its Target Organs
Poelaert, K. C. K., Cleemput, J. V., Laval, K., Favoreel, H. W., Couck, L., Broeck, W. V. D., Azab, W., & Nauwynck, H. (2019) · Journal of Virology · doi:10.1128/jvi.02098-18 · Cited by 33 · see citing works
“(88) previously reported the isolation and immortalization of equine venous endothelial cells (EC).”
BackgroundSelf-citationMarinho, R. D. S. S., Ramos, C. J. B., Leite, J. P. G., Teixeira, V. L., Paixão, I. C. N. D. P., Belo, C. A. D., Pereira, A. B., & Pinto, A. M. V. (2016) · Journal of Applied Phycology · doi:10.1007/s10811-016-0946-9 · Cited by 32 · see citing works
“A member of the subfamily Alphaherpesvirinae, equine herpesvirus 1 (EHV-1) is a major pathogen of horses worldwide and can cause of respiratory disease, abortion in pregnant mares, perinatal foal mortality, and myeloencephalopathy in horses, even vaccinated ones (Laval et al., 2015).”
BackgroundEquine Herpesvirus-1 Myeloencephalopathy
Pusterla, N., Hussey, G. S., & Goehring, L. S. (2022) · Veterinary Clinics of North America Equine Practice · doi:10.1016/j.cveq.2022.05.006 · Cited by 30 · see citing works
Zhao, J., Poelaert, K. C. K., Van Cleemput, J., & Nauwynck, H. J. (2017) · Veterinary Research · doi:10.1186/s13567-017-0419-4 · Cited by 23 · see citing works
Self-citationZarski, L. M., Giessler, K. S., Jacob, S. I., Weber, P. S. D., McCauley, A. G., Lee, Y., & Hussey, G. S. (2021) · Viruses · doi:10.3390/v13030356 · Cited by 21 · see citing works
“thus enhances cell adhesion and viral trans-fer with the vascular endothelial cells [98,99].”
BackgroundEndothelial cells and angiogenesis in the horse in health and disease—A review
Rieger, J., Kaessmeyer, S., Masri, S. A., Hünigen, H., & Plendl, J. (2020) · Anatomia Histologia Embryologia · doi:10.1111/ahe.12588 · Cited by 18 · see citing works
“Mechanism by which equine herpesvirus type 1 is transferred from CD172a + cells to endothelial cells Vena cava Laval et al. (2015) Equine Herpesvirus Type 1 Enhances Viral Replication in CD172a(+) Monocytic Cells upon Adhesion to Endothelial Cells.”
BackgroundPoelaert, K. C. K., Cleemput, J. V., Laval, K., Descamps, S., Favoreel, H. W., & Nauwynck, H. (2019) · Frontiers in Microbiology · doi:10.3389/fmicb.2019.00723 · Cited by 18 · see citing works
Self-citationHussey, G. S., & Giessler, K. S. (2022) · The Veterinary Journal · doi:10.1016/j.tvjl.2022.105827 · Cited by 16 · see citing works
Replication of neurovirulent equine herpesvirus type 1 (EHV-1) in CD172a(+) monocytic cells.
Laval, K., Van Cleemput, J., Poelaert, K. C., Brown, I. K., & Nauwynck, H. J. (2017) · Comparative Immunology Microbiology and Infectious Diseases · doi:10.1016/j.cimid.2016.11.006 · Cited by 14 · see citing works
“EHV-1 gB protein was exclusively expressed in the cytoplasm of CD172a+ cells, as previously described (13) (data not shown).”
Pavulraj, S., Kamel, M., Stephanowitz, H., Liu, F., Plendl, J., Osterrieder, N., & Azab, W. (2020) · Viruses · doi:10.3390/v12090999 · Cited by 13 · see citing works
“Herpesvirus infection pathway is mediated by numerous host and viral proteins which includes several pathways such as toll-like receptor signaling, pro-inflammatory cytokines, inhibition of apoptosis, antigen processing and presentation, JAK-STAT, calcium signaling, nuclear factor-kappa B (NF-κB), and B cell receptor…”
BackgroundLiu, S. A., Stanfield, B. A., Chouljenko, V. N., Naidu, S. K., Langohr, I. M., Piero, F. D., Ferracone, J., Roy, A. A., & Kousoulas, K. G. (2017) · Journal of Virology · doi:10.1128/jvi.02445-16 · Cited by 13 · see citing works
Entry of equid herpesvirus 1 into CD172a+ monocytic cells.
Laval, K., Favoreel, H. W., Van Cleemput, J., Poelaert, K. C. K., Brown, I. K., Verhasselt, B., & Nauwynck, H. J. (2016) · Journal of General Virology · doi:10.1099/jgv.0.000375 · Cited by 12 · see citing works
Zarski, L. M., Weber, P. S. D., Lee, Y., & Hussey, G. S. (2021) · Pathogens · doi:10.3390/pathogens10010043 · Cited by 12 · see citing works
Anti‐inflammatory drugs decrease infection of brain endothelial cells with EHV ‐1 in vitro
Goehring, L. S., Brandes, K., Ashton, L. V., Wittenburg, L. A., Olea‐Popelka, F. J., Lunn, D. P., & Hussey, G. S. (2016) · Equine Veterinary Journal · doi:10.1111/evj.12656 · Cited by 12 · see citing works
“Two independent, recent publications also suggest that it is indeed the infection of PBMC with EHV-1 that increases the likelihood of interaction with EC in in vitro models using EC of non-CNS origin [24,25].”
BackgroundEven, K. M., Gaesser, A. M., Ciamillo, S. A., Linardi, R. L., & Ortved, K. F. (2022) · Frontiers in Veterinary Science · doi:10.3389/fvets.2022.958724 · Cited by 11 · see citing works
Characterization of feline herpesvirus-1 deletion mutants in tissue explant cultures
Lee, Y., Maes, R. K., Kiupel, M., Nauwynck, H., & Hussey, G. S. (2020) · Virus Research · doi:10.1016/j.virusres.2020.197981 · Cited by 9 · see citing works
Self-citationEHV-1 Pathogenesis: Current in vitro Models and Future Perspectives
Kamel, M. S., Pavulraj, S., Osterrieder, N., & Azab, W. (2019) · Frontiers in Veterinary Science · doi:10.3389/fvets.2019.00251 · Cited by 9 · see citing works
“The system proved to be flexible to study other aspects during virus spread, particularly the role of adhesion molecules in virus transmission (27).”
BackgroundZheng, L., Byadgi, O. V., Rakhshaninejad, M., & Nauwynck, H. (2022) · Fish & Shellfish Immunology · doi:10.1016/j.fsi.2022.08.021 · Cited by 7 · see citing works
Self-citationKamel, M., Pavulraj, S., Fauler, B., Mielke, T., & Azab, W. (2020) · iScience · doi:10.1016/j.isci.2020.101615 · Cited by 7 · see citing works
Crombrugge, E. V., Vanbeylen, E., Cleemput, J. V., Broeck, W. V. D., Laval, K., & Nauwynck, H. (2022) · Viruses · doi:10.3390/v14010149 · Cited by 7 · see citing works
“After washing the sections three times in PBS, an immunofluorescent staining to detect EHV-1 late proteins was performed, as previously described [29].”
MethodSelf-citationMohamed, E., Žarak, I., Vereecke, N., Theuns, S., Laval, K., & Nauwynck, H. (2025) · Veterinary Research · doi:10.1186/s13567-024-01434-3 · Cited by 4 · see citing works
Self-citationPeptide Mediated Adhesion to Beta-Lactam Ring of Equine Mesenchymal Stem Cells: A Pilot Study
Merlo, B., Baldassarro, V. A., Flagelli, A., Marcoccia, R., Giraldi, V., Focarete, M. L., Giacomini, D., & Iacono, E. (2022) · Animals · doi:10.3390/ani12060734 · Cited by 4 · see citing works
Equine Herpesvirus Infections: Treatment Progress and Challenges in Horses and Donkeys
Khan, M. Z., Ji, Y., Fan, X., Liu, Y., Liu, W., & Wang, C. (2025) · Veterinary Sciences · doi:10.3390/vetsci12111082 · Cited by 5 · see citing works
Equine histones are mobilized within equid alphaherpesvirus 1 (EHV1) replication compartments
Conn, K. L. (2025) · Journal of Virology · doi:10.1128/jvi.01589-25 · Cited by 3 · see citing works
“It estab lishes primary lytic replication within epithelial cells of the upper respiratory tract, disseminates throughout the body in a monocyte-associated viremia, and establishes secondary lytic replication in endothelial cells lining blood vessels (3–5).”
Meeremans, M., Devriendt, B., Bairiot, S., Poucke, M. V., Peelman, L. J., Demeyere, K., Meyer, E., Vlierberghe, S. V., & Schauwer, C. D. (2025) · Veterinary Quarterly · doi:10.1080/01652176.2025.2593367 · Cited by 3 · see citing works
“In the current study, equine monocytes were isolated from peripheral blood (PB) based on surface expression of the marker CD172a (Baghi et al. 2014; Laval et al. 2015; Claessen et al. 2016) and differentiated into macrophages.”
Minato, E., Aoshima, K., Kobayashi, A., Ohnishi, N., Sasaki, N., & Kimura, T. (2019) · Veterinary Pathology · doi:10.1177/0300985819834616 · Cited by 1 · see citing works
Partiot, E., Brychka, D., & Gaudin, R. (2024) · European Journal of Cell Biology · doi:10.1016/j.ejcb.2024.151453 · Cited by 1 · see citing works
Lascola, K. M., Wilkins, P. A., Wong, D. M., et al. (2024) · doi:10.1002/9781119617228.ch8 · Cited by 1 · see citing works
Lessiak, U., Melchert, M., Walter, I., Kummer, S., Nell, B., Tschulenk, W., & Pratscher, B. (2024) · Frontiers in Veterinary Science · doi:10.3389/fvets.2024.1421946
Stellungnahme zur Wirksamkeit und immunologischen Sicherheit von EHV-Impfstoffen bei Pferden
Vet), A. P. D. S. I. V. (. (2025) · Tierärztliche Praxis Ausgabe G Großtiere / Nutztiere · doi:10.1055/a-2518-1976
“EHV moduliert die Expression von Adhäsionsmolekülen auf infizierten, mononukleären Zellen [25, 26].”
Conn, K. L. (2025) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2025.08.01.668067
Pokholenko, I., Meeremans, M., Vlierberghe, S. V., Pien, N., & Schauwer, C. D. (2026) · Frontiers in Bioengineering and Biotechnology · doi:10.3389/fbioe.2026.1677364
Quentin-Froignant, C. (2021) · doi:10.70675/eda0a1edzb032z43b0za6d4z83d3f85db887
Tishchenko, A., Boeck, B. W., Waesberghe, C. V., et al. (2026) · Journal of Virology · doi:10.1128/jvi.00773-26
“…been documented for a wide range of DNA (33) and RNA viruses (34), including several herpesviruses such as HSV-1 (35, 36), HSV-2 (37), VZV (38), EHV-1 (39), BoHV-1 (40), KSHV (41), HCMV (42, 43), and EBV (44); and is commonly associated with enhanced viral spread and replication (42, 43, 45–51).”
InfluentialSelf-citation
- Authors
- Laval, K., Favoreel, H. W., Poelaert, K. C. K., Van Cleemput, J., & Nauwynck, H. J.
- Affiliations
- Ghent University (BE)
- Department of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium
- Venue
- Journal of Virology
- Publisher
- American Society for Microbiology
- Published
- 2015-11
- Volume
- 89
- Issue
- 21
- Pages
- 10912-10923
- ISSN
- 0022-538X (ISSN-L)
- 1098-5514
- PMC
- PMC4621108
- OpenAlex
- W1645406539
- Cited by
- 43
- References
- 50
- Citations by year
- 2026: 2 · 2025: 6 · 2024: 3 · 2023: 1 · 2022: 6 · 2021: 4 · 2020: 5 · 2019: 6 · 2018: 3 · 2017: 3 · 2016: 3 · 2015: 1
- FWCI
- 2.05
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Virology (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (submittedVersion)
- Ghent University Academic Bibliography (Ghent University) (submittedVersion)
- Topics
- Health Sciences › Medicine › Epidemiology › Herpesvirus Infections and Treatments
- Life Sciences › Agricultural and Biological Sciences › Animal Science and Zoology › Animal Virus Infections Studies
- Health Sciences › Medicine › Surgery › Xenotransplantation and immune response
- Keywords
- endothelial cells, cell adhesion, integrins, integrin αvβ3, PI3K signaling, ERK signaling, viral replication, Trojan horse
- MeSH terms
- Analysis of Variance, Animals, Belgium, Cell Adhesion, Enzyme-Linked Immunosorbent Assay, Herpesvirus 1, Equid, Flow Cytometry, Herpesviridae Infections
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Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
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