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Article ; Online: Quantification of camelid cytokine mRNA expression in PBMCs by microfluidic qPCR technology.

Rodon, Jordi / Te, Nigeer / Ballester, Maria / Segalés, Joaquim / Vergara-Alert, Júlia / Bensaid, Albert

Developmental and comparative immunology

2023 Volume 149, Page(s) 105061

Abstract: Camelids are economically and socially important in several parts of the world and might carry pathogens with epizootic or zoonotic potential. However, biological research in these species is limited due to lack of reagents. Here, we developed RT-qPCR ... ...

Abstract	Camelids are economically and socially important in several parts of the world and might carry pathogens with epizootic or zoonotic potential. However, biological research in these species is limited due to lack of reagents. Here, we developed RT-qPCR assays to quantify a panel of camelid innate and adaptive immune response genes, which can be monitored in a single run. The assays were validated with PHA, PMA-ionomycin, and Poly I:C-stimulated PBMCs from alpaca, dromedary camel and llama, including normalization by multiple reference genes. Further, comparative gene expression analyses for the different camelid species were performed by a unique microfluidic qPCR assay. Compared to unstimulated controls, PHA and PMA-ionomycin stimulation elicited robust Th1 and Th2 responses in PBMCs from camelid species. Additional activation of type I and type III IFN signalling pathways was described exclusively in PHA-stimulated dromedary lymphocytes, in contrast to those from alpaca and llama. We also found that PolyI:C stimulation induced robust antiviral response genes in alpaca PBMCs. The proposed methodology should be useful for the measurement of immune responses to infection or vaccination in camelid species.
MeSH term(s)	Animals ; Cytokines/genetics ; Camelids, New World ; Camelus ; Ionomycin ; Microfluidics ; RNA, Messenger
Chemical Substances	Cytokines ; Ionomycin (56092-81-0) ; RNA, Messenger
Language	English
Publishing date	2023-09-17
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	752411-0
ISSN	1879-0089 ; 0145-305X
ISSN (online)	1879-0089
ISSN	0145-305X
DOI	10.1016/j.dci.2023.105061
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Article ; Online: Enhanced antiviral immunity and dampened inflammation in llama lymph nodes upon MERS-CoV sensing: bridging innate and adaptive cellular immune responses in camelid reservoirs.

Rodon, Jordi / Te, Nigeer / Segalés, Joaquim / Vergara-Alert, Júlia / Bensaid, Albert

Frontiers in immunology

2023 Volume 14, Page(s) 1205080

Abstract: Middle East respiratory syndrome coronavirus (MERS-CoV) infection can cause fatal pulmonary inflammatory disease in humans. Contrarily, camelids and bats are the main reservoir hosts, tolerant for MERS-CoV replication without suffering clinical disease. ... ...

Abstract	Middle East respiratory syndrome coronavirus (MERS-CoV) infection can cause fatal pulmonary inflammatory disease in humans. Contrarily, camelids and bats are the main reservoir hosts, tolerant for MERS-CoV replication without suffering clinical disease. Here, we isolated cervical lymph node (LN) cells from MERS-CoV convalescent llamas and pulsed them with two different viral strains (clades B and C). Viral replication was not supported in LN, but a cellular immune response was mounted. Reminiscent Th1 responses (IFN-γ, IL-2, IL-12) were elicited upon MERS-CoV sensing, accompanied by a marked and transient peak of antiviral responses (type I IFNs, IFN-λ3, ISGs, PRRs and TFs). Importantly, expression of inflammatory cytokines (TNF-α, IL-1β, IL-6, IL-8) or inflammasome components (NLRP3, CASP1, PYCARD) was dampened. The role of IFN-λ3 to counterbalance inflammatory processes and bridge innate and adaptive immune responses in camelid species is discussed. Our findings shed light into key mechanisms on how reservoir species control MERS-CoV in the absence of clinical disease.
MeSH term(s)	Humans ; Animals ; Antiviral Agents ; Camelids, New World ; Middle East Respiratory Syndrome Coronavirus ; Camelidae ; Chiroptera ; Inflammation ; Immunity, Cellular
Chemical Substances	Antiviral Agents
Language	English
Publishing date	2023-06-14
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2023.1205080
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Middle East respiratory coronavirus (MERS-CoV) internalized by llama alveolar macrophages does not result in virus replication or induction of pro-inflammatory cytokines.

Rodon, Jordi / Sachse, Martin / Te, Nigeer / Segalés, Joaquim / Bensaid, Albert / Risco, Cristina / Vergara-Alert, Júlia

Microbes and infection

2023 Volume 26, Issue 3, Page(s) 105252

Abstract: Severe Middle East respiratory syndrome (MERS) is characterized by massive infiltration of immune cells in lungs. MERS-coronavirus (MERS-CoV) replicates in vitro in human macrophages, inducing high pro-inflammatory responses. In contrast, camelids, the ... ...

Abstract	Severe Middle East respiratory syndrome (MERS) is characterized by massive infiltration of immune cells in lungs. MERS-coronavirus (MERS-CoV) replicates in vitro in human macrophages, inducing high pro-inflammatory responses. In contrast, camelids, the main reservoir for MERS-CoV, are asymptomatic carriers. Although limited infiltration of leukocytes has been observed in the lower respiratory tract of camelids, their role during infection remains unknown. Here we studied whether llama alveolar macrophages (LAMs) are susceptible to MERS-CoV infection and can elicit pro-inflammatory responses. MERS-CoV did not replicate in LAMs; however, they effectively capture and degrade viral particles. Moreover, transcriptomic analyses showed that LAMs do not induce pro-inflammatory cytokines upon MERS-CoV sensing.
MeSH term(s)	Animals ; Humans ; Cytokines/metabolism ; Middle East Respiratory Syndrome Coronavirus ; Macrophages, Alveolar ; Camelids, New World/metabolism ; Virus Replication ; Coronavirus Infections
Chemical Substances	Cytokines
Language	English
Publishing date	2023-11-18
Publishing country	France
Document type	Journal Article
ZDB-ID	1465093-9
ISSN	1769-714X ; 1286-4579
ISSN (online)	1769-714X
ISSN	1286-4579
DOI	10.1016/j.micinf.2023.105252
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Article ; Online: Enhanced replication fitness of MERS-CoV clade B over clade A strains in camelids explains the dominance of clade B strains in the Arabian Peninsula.

Te, Nigeer / Rodon, Jordi / Pérez, Mónica / Segalés, Joaquim / Vergara-Alert, Júlia / Bensaid, Albert

Emerging microbes & infections

2021 Volume 11, Issue 1, Page(s) 260–274

Abstract: ... ...

Abstract	ABSTRACT
MeSH term(s)	Adaptation, Physiological/genetics ; Amino Acid Substitution/genetics ; Animals ; Camelids, New World ; Camelus ; Cell Line ; Chlorocebus aethiops ; Coronavirus Infections/epidemiology ; Coronavirus Infections/veterinary ; Cytokines/blood ; Genome, Viral/genetics ; Immunity, Innate/immunology ; Jordan/epidemiology ; Middle East Respiratory Syndrome Coronavirus/classification ; Middle East Respiratory Syndrome Coronavirus/genetics ; Open Reading Frames/genetics ; Qatar/epidemiology ; RNA, Viral/genetics ; Respiratory Mucosa/virology ; Spike Glycoprotein, Coronavirus/genetics ; Vero Cells ; Viral Load
Chemical Substances	Cytokines ; RNA, Viral ; Spike Glycoprotein, Coronavirus
Language	English
Publishing date	2021-12-17
Publishing country	United States
Document type	Journal Article
ZDB-ID	2681359-2
ISSN	2222-1751 ; 2222-1751
ISSN (online)	2222-1751
ISSN	2222-1751
DOI	10.1080/22221751.2021.2019559
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Evaluation of alpaca tracheal explants as an ex vivo model for the study of Middle East respiratory syndrome coronavirus (MERS-CoV) infection

Te, Nigeer / Rodon, Jordi / Creve, Rhea / Pérez, Mónica / Segalés, Joaquim / Vergara-Alert, Júlia / Bensaid, Albert

BMC veterinary research. 2022 Dec., v. 53, no. 1

2022

Abstract: Middle East respiratory syndrome coronavirus (MERS-CoV) poses a serious threat to public health. Here, we established an ex vivo alpaca tracheal explant (ATE) model using an air-liquid interface culture system to gain insights into MERS-CoV infection in ... ...

Abstract	Middle East respiratory syndrome coronavirus (MERS-CoV) poses a serious threat to public health. Here, we established an ex vivo alpaca tracheal explant (ATE) model using an air-liquid interface culture system to gain insights into MERS-CoV infection in the camelid lower respiratory tract. ATE can be infected by MERS-CoV, being 10³ TCID₅₀/mL the minimum viral dosage required to establish a productive infection. IFNs and antiviral ISGs were not induced in ATE cultures in response to MERS-CoV infection, strongly suggesting that ISGs expression observed in vivo is rather a consequence of the IFN induction occurring in the nasal mucosa of camelids.
Keywords	Camelidae ; Coronavirus infections ; Middle East respiratory syndrome coronavirus ; alpacas ; ex vivo studies ; liquid-air interface ; models ; nasal mucosa ; public health ; veterinary medicine
Language	English
Dates of publication	2022-12
Size	p. 67.
Publishing place	BioMed Central
Document type	Article
ZDB-ID	1146298-x
ISSN	1297-9716 ; 0928-4249
ISSN (online)	1297-9716
ISSN	0928-4249
DOI	10.1186/s13567-022-01084-3
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Evaluation of alpaca tracheal explants as an ex vivo model for the study of Middle East respiratory syndrome coronavirus (MERS-CoV) infection.

Te, Nigeer / Rodon, Jordi / Creve, Rhea / Pérez, Mónica / Segalés, Joaquim / Vergara-Alert, Júlia / Bensaid, Albert

Veterinary research

2022 Volume 53, Issue 1, Page(s) 67

Abstract	Middle East respiratory syndrome coronavirus (MERS-CoV) poses a serious threat to public health. Here, we established an ex vivo alpaca tracheal explant (ATE) model using an air-liquid interface culture system to gain insights into MERS-CoV infection in the camelid lower respiratory tract. ATE can be infected by MERS-CoV, being 10
MeSH term(s)	Animals ; Antiviral Agents ; Bronchi ; Camelids, New World ; Coronavirus Infections/veterinary ; Middle East Respiratory Syndrome Coronavirus/physiology
Chemical Substances	Antiviral Agents
Language	English
Publishing date	2022-09-02
Publishing country	England
Document type	Journal Article
ZDB-ID	1146298-x
ISSN	1297-9716 ; 0928-4249
ISSN (online)	1297-9716
ISSN	0928-4249
DOI	10.1186/s13567-022-01084-3
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Article ; Online: Evaluation of alpaca tracheal explants as an ex vivo model for the study of Middle East respiratory syndrome coronavirus (MERS-CoV) infection

Nigeer Te / Jordi Rodon / Rhea Creve / Mónica Pérez / Joaquim Segalés / Júlia Vergara-Alert / Albert Bensaid

Veterinary Research, Vol 53, Iss 1, Pp 1-

2022 Volume 7

Abstract: Abstract Middle East respiratory syndrome coronavirus (MERS-CoV) poses a serious threat to public health. Here, we established an ex vivo alpaca tracheal explant (ATE) model using an air-liquid interface culture system to gain insights into MERS-CoV ... ...

Abstract	Abstract Middle East respiratory syndrome coronavirus (MERS-CoV) poses a serious threat to public health. Here, we established an ex vivo alpaca tracheal explant (ATE) model using an air-liquid interface culture system to gain insights into MERS-CoV infection in the camelid lower respiratory tract. ATE can be infected by MERS-CoV, being 103 TCID50/mL the minimum viral dosage required to establish a productive infection. IFNs and antiviral ISGs were not induced in ATE cultures in response to MERS-CoV infection, strongly suggesting that ISGs expression observed in vivo is rather a consequence of the IFN induction occurring in the nasal mucosa of camelids.
Keywords	Air-liquid interface ; alpaca ; camelid ; ex vivo model ; MERS-CoV ; tracheal explants ; Veterinary medicine ; SF600-1100
Language	English
Publishing date	2022-09-01T00:00:00Z
Publisher	BMC
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Extended Viral Shedding of MERS-CoV Clade B Virus in Llamas Compared with African Clade C Strain.

Rodon, Jordi / Mykytyn, Anna Z / Te, Nigeer / Okba, Nisreen M A / Lamers, Mart M / Pailler-García, Lola / Cantero, Guillermo / Albulescu, Irina / Bosch, Berend-Jan / Peiris, Malik / Bensaid, Albert / Vergara-Alert, Júlia / Haagmans, Bart L / Segalés, Joaquim

Emerging infectious diseases

2023 Volume 29, Issue 3, Page(s) 585–589

Abstract: Middle East respiratory syndrome coronavirus (MERS-CoV) clade B viruses are found in camelids and humans in the Middle East, but clade C viruses are not. We provide experimental evidence for extended shedding of MERS-CoV clade B viruses in llamas, which ... ...

Abstract	Middle East respiratory syndrome coronavirus (MERS-CoV) clade B viruses are found in camelids and humans in the Middle East, but clade C viruses are not. We provide experimental evidence for extended shedding of MERS-CoV clade B viruses in llamas, which might explain why they outcompete clade C strains in the Arabian Peninsula.
MeSH term(s)	Animals ; Humans ; Middle East Respiratory Syndrome Coronavirus ; Camelids, New World ; Herpesvirus 1, Cercopithecine ; Coronavirus Infections ; Virus Shedding ; Camelus
Language	English
Publishing date	2023-02-16
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1380686-5
ISSN	1080-6059 ; 1080-6040
ISSN (online)	1080-6059
ISSN	1080-6040
DOI	10.3201/eid2903.220986
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Article ; Online: Type I and III IFNs produced by the nasal epithelia and dimmed inflammation are features of alpacas resolving MERS-CoV infection.

Te, Nigeer / Rodon, Jordi / Ballester, Maria / Pérez, Mónica / Pailler-García, Lola / Segalés, Joaquim / Vergara-Alert, Júlia / Bensaid, Albert

PLoS pathogens

2021 Volume 17, Issue 5, Page(s) e1009229

Abstract: While MERS-CoV (Middle East respiratory syndrome Coronavirus) provokes a lethal disease in humans, camelids, the main virus reservoir, are asymptomatic carriers, suggesting a crucial role for innate immune responses in controlling the infection. ... ...

Abstract	While MERS-CoV (Middle East respiratory syndrome Coronavirus) provokes a lethal disease in humans, camelids, the main virus reservoir, are asymptomatic carriers, suggesting a crucial role for innate immune responses in controlling the infection. Experimentally infected camelids clear infectious virus within one week and mount an effective adaptive immune response. Here, transcription of immune response genes was monitored in the respiratory tract of MERS-CoV infected alpacas. Concomitant to the peak of infection, occurring at 2 days post inoculation (dpi), type I and III interferons (IFNs) were maximally transcribed only in the nasal mucosa of alpacas, while interferon stimulated genes (ISGs) were induced along the whole respiratory tract. Simultaneous to mild focal infiltration of leukocytes in nasal mucosa and submucosa, upregulation of the anti-inflammatory cytokine IL10 and dampened transcription of pro-inflammatory genes under NF-κB control were observed. In the lung, early (1 dpi) transcription of chemokines (CCL2 and CCL3) correlated with a transient accumulation of mainly mononuclear leukocytes. A tight regulation of IFNs in lungs with expression of ISGs and controlled inflammatory responses, might contribute to virus clearance without causing tissue damage. Thus, the nasal mucosa, the main target of MERS-CoV in camelids, seems central in driving an efficient innate immune response based on triggering ISGs as well as the dual anti-inflammatory effects of type III IFNs and IL10.
MeSH term(s)	Animals ; Antiviral Agents/metabolism ; Antiviral Agents/pharmacology ; Camelids, New World/immunology ; Camelids, New World/metabolism ; Camelids, New World/virology ; Chlorocebus aethiops ; Coronavirus Infections/immunology ; Coronavirus Infections/metabolism ; Coronavirus Infections/prevention & control ; Coronavirus Infections/veterinary ; Disease Reservoirs/veterinary ; Disease Resistance/drug effects ; Disease Resistance/genetics ; Disease Resistance/immunology ; Gene Expression Regulation ; Immunity, Innate/physiology ; Inflammation/immunology ; Inflammation/metabolism ; Inflammation/veterinary ; Inflammation/virology ; Interferon Type I/genetics ; Interferon Type I/metabolism ; Interferon Type I/pharmacology ; Interferons/genetics ; Interferons/metabolism ; Interferons/pharmacology ; Middle East Respiratory Syndrome Coronavirus/drug effects ; Middle East Respiratory Syndrome Coronavirus/immunology ; Middle East Respiratory Syndrome Coronavirus/physiology ; Nasal Mucosa/drug effects ; Nasal Mucosa/immunology ; Nasal Mucosa/metabolism ; Nasal Mucosa/virology ; Respiratory System/drug effects ; Respiratory System/immunology ; Respiratory System/metabolism ; Respiratory System/virology ; Vero Cells ; Viral Load/drug effects ; Virus Replication/drug effects
Chemical Substances	Antiviral Agents ; Interferon Type I ; interferon type III ; Interferons (9008-11-1)
Language	English
Publishing date	2021-05-24
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2205412-1
ISSN	1553-7374 ; 1553-7366
ISSN (online)	1553-7374
ISSN	1553-7366
DOI	10.1371/journal.ppat.1009229
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Article ; Online: Middle East respiratory syndrome coronavirus infection in camelids.

Te, Nigeer / Ciurkiewicz, Malgorzata / van den Brand, Judith M A / Rodon, Jordi / Haverkamp, Ann-Kathrin / Vergara-Alert, Júlia / Bensaid, Albert / Haagmans, Bart L / Baumgartner, Wolfgang / Segalés, Joaquim

Veterinary pathology

2022 Volume 59, Issue 4, Page(s) 546–555

Abstract: Middle East respiratory syndrome coronavirus (MERS-CoV) is the cause of a severe respiratory disease with a high case fatality rate in humans. Since its emergence in mid-2012, 2578 laboratory-confirmed cases in 27 countries have been reported by the ... ...

Abstract	Middle East respiratory syndrome coronavirus (MERS-CoV) is the cause of a severe respiratory disease with a high case fatality rate in humans. Since its emergence in mid-2012, 2578 laboratory-confirmed cases in 27 countries have been reported by the World Health Organization, leading to 888 known deaths due to the disease and related complications. Dromedary camels are considered the major reservoir host for this virus leading to zoonotic infection in humans. Dromedary camels, llamas, and alpacas are susceptible to MERS-CoV, developing a mild-to-moderate upper respiratory tract infection characterized by epithelial hyperplasia as well as infiltration of neutrophils, lymphocytes, and some macrophages within epithelium, lamina propria, in association with abundant viral antigen. The very mild lesions in the lower respiratory tract of these camelids correlate with absence of overt illness following MERS-CoV infection. Unfortunately, there is no approved antiviral treatment or vaccine for MERS-CoV infection in humans. Thus, there is an urgent need to develop intervention strategies in camelids, such as vaccination, to minimize virus spillover to humans. Therefore, the development of camelid models of MERS-CoV infection is key not only to assess vaccine prototypes but also to understand the biologic mechanisms by which the infection can be naturally controlled in these reservoir species. This review summarizes information on virus-induced pathological changes, pathogenesis, viral epidemiology, and control strategies in camelids, as the intermediate hosts and primary source of MERS-CoV infection in humans.
MeSH term(s)	Animals ; Camelids, New World ; Camelus ; Coronavirus Infections/epidemiology ; Coronavirus Infections/veterinary ; Middle East Respiratory Syndrome Coronavirus ; Zoonoses
Language	English
Publishing date	2022-01-08
Publishing country	United States
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't
ZDB-ID	188012-3
ISSN	1544-2217 ; 0300-9858
ISSN (online)	1544-2217
ISSN	0300-9858
DOI	10.1177/03009858211069120
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