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Article ; Online: Comprehensive Analysis of Soluble Mediator Profiles in Congenital CMV Infection Using an MCMV Model.

Karner, Dubravka / Kvestak, Daria / Lisnic, Berislav / Cokaric Brdovcak, Maja / Juranic Lisnic, Vanda / Kucan Brlic, Paola / Hasan, Milena / Lenac Rovis, Tihana

Viruses

2024 Volume 16, Issue 2

Abstract: Congenital human cytomegalovirus (HCMV) infection may cause life-threatening disease and permanent damage to the central nervous system. The mouse model of CMV infection is most commonly used to study mechanisms of infection and pathogenesis. While ... ...

Abstract	Congenital human cytomegalovirus (HCMV) infection may cause life-threatening disease and permanent damage to the central nervous system. The mouse model of CMV infection is most commonly used to study mechanisms of infection and pathogenesis. While essential to limit mouse CMV (MCMV) replication, the inflammatory responses, particularly IFNγ and TNFα, cause neurodevelopmental abnormalities. Other soluble mediators of the immune response in most tissues remain largely unexplored. To address this gap, we quantified 48 soluble mediators of the immune response, including 32 cytokines, 10 chemokines, 3 growth factors/regulators, and 3 soluble receptors in the spleen, liver, lungs, and brain at 9 and 14 days postinfection (dpi). Our analysis found 25 induced molecules in the brain at 9 dpi, with an additional 8 showing statistically elevated responses at 14 dpi. Specifically, all analyzed CCL group cytokines (CCL2, CCL3, CCL4, CCL5, CCL7, and CCL11) were upregulated at 14 dpi in the brain. Furthermore, data revealed differentially regulated analytes across tissues, such as CCL11, CXCL5, and IL-10 in the brain, IL-33/IL-33R in the liver, and VEGF-a and IL-5 in the lungs. Overall, this study provides an overview of the immune dynamics of soluble mediators in congenital CMV.
MeSH term(s)	Animals ; Humans ; Mice ; Cytomegalovirus Infections ; Cytokines ; Brain ; Tumor Necrosis Factor-alpha ; Muromegalovirus
Chemical Substances	Cytokines ; Tumor Necrosis Factor-alpha
Language	English
Publishing date	2024-01-30
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v16020208
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Mouse Models for Cytomegalovirus Infections in Newborns and Adults.

Brizić, Ilija / Lisnić, Berislav / Krstanović, Fran / Brune, Wolfram / Hengel, Hartmut / Jonjić, Stipan

Current protocols

2022 Volume 2, Issue 9, Page(s) e537

Abstract: This article describes procedures for infecting adult mice with murine cytomegalovirus (MCMV) and for infecting newborn mice to model congenital CMV infection. Methods are included for propagating MCMV in cell cultures and preparing a more virulent form ... ...

Abstract	This article describes procedures for infecting adult mice with murine cytomegalovirus (MCMV) and for infecting newborn mice to model congenital CMV infection. Methods are included for propagating MCMV in cell cultures and preparing a more virulent form of MCMV from the salivary glands of infected mice. A plaque assay is provided for determining MCMV titers of infected tissues or virus stocks. Also, methods are described for preparing the murine embryonic fibroblasts used for propagating MCMV, and for the plaque assay. © 2022 Wiley Periodicals LLC.
MeSH term(s)	Animals ; Cytomegalovirus Infections ; Disease Models, Animal ; Mice ; Muromegalovirus ; Salivary Glands
Language	English
Publishing date	2022-09-09
Publishing country	United States
Document type	Journal Article
ISSN	2691-1299
ISSN (online)	2691-1299
DOI	10.1002/cpz1.537
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Article ; Online: Rodent Models of Congenital Cytomegalovirus Infection.

Lisnić, Berislav / Tomac, Jelena / Cekinović, Djurdjica / Jonjić, Stipan / Juranić Lisnić, Vanda

Methods in molecular biology (Clifton, N.J.)

2021 Volume 2244, Page(s) 365–401

Abstract: Human cytomegalovirus (HCMV) is a leading viral cause of congenital infections in the central nervous system (CNS) and may result in severe long-term sequelae. High rates of sequelae following congenital HCMV infection and insufficient antiviral therapy ... ...

Abstract	Human cytomegalovirus (HCMV) is a leading viral cause of congenital infections in the central nervous system (CNS) and may result in severe long-term sequelae. High rates of sequelae following congenital HCMV infection and insufficient antiviral therapy in the perinatal period makes the development of an HCMV-specific vaccine a high priority of modern medicine. Due to the species specificity of HCMV, animal models are frequently used to study CMV pathogenesis. Studies of murine cytomegalovirus (MCMV) infections of adult mice have played a significant role as a model of CMV biology and pathogenesis, while MCMV infection of newborn mice has been successfully used as a model of perinatal CMV infection. Newborn mice infected with MCMV have high levels of viremia during which the virus establishes a productive infection in most organs, coupled with a robust inflammatory response. Productive infection in the brain parenchyma during early postnatal period leads to an extensive nonnecrotizing multifocal widespread encephalitis characterized by infiltration of components of both innate and adaptive immunity. As a result, impairment in postnatal development of mouse cerebellum leads to long-term motor and sensor disabilities. This chapter summarizes current findings of rodent models of perinatal CMV infection and describes methods for analysis of perinatal MCMV infection in newborn mice.
MeSH term(s)	Animals ; Animals, Newborn ; Brain/immunology ; Central Nervous System/virology ; Cytomegalovirus/immunology ; Cytomegalovirus/metabolism ; Cytomegalovirus/pathogenicity ; Cytomegalovirus Infections/immunology ; Disease Models, Animal ; Encephalitis ; Fetal Diseases ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Muromegalovirus/immunology ; Primary Cell Culture
Language	English
Publishing date	2021-02-08
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-1111-1_18
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Article ; Online: Membraneless Compartmentalization of Nuclear Assembly Sites during Murine Cytomegalovirus Infection.

Mahmutefendić Lučin, Hana / Lukanović Jurić, Silvija / Marcelić, Marina / Štimac, Igor / Viduka, Ivona / Blagojević Zagorac, Gordana / Lisnić, Berislav / Ruzsics, Zsolt / Lučin, Pero

Viruses

2023 Volume 15, Issue 3

Abstract: Extensive reorganization of infected cells and the formation of large structures known as the nuclear replication compartment (RC) and cytoplasmic assembly compartment (AC) is a hallmark of beta-herpesvirus infection. These restructurings rely on ... ...

Abstract	Extensive reorganization of infected cells and the formation of large structures known as the nuclear replication compartment (RC) and cytoplasmic assembly compartment (AC) is a hallmark of beta-herpesvirus infection. These restructurings rely on extensive compartmentalization of the processes that make up the virus manufacturing chain. Compartmentalization of the nuclear processes during murine cytomegalovirus (MCMV) infection is not well described. In this study, we visualized five viral proteins (pIE1, pE1, pM25, pm48.2, and pM57) and replicated viral DNA to reveal the nuclear events during MCMV infection. As expected, these events can be matched with those described for other beta and alpha herpesviruses and contribute to the overall picture of herpesvirus assembly. Imaging showed that four viral proteins (pE1, pM25, pm48.2, and pM57) and replicated viral DNA condense in the nucleus into membraneless assemblies (MLAs) that undergo a maturation sequence to form the RC. One of these proteins (pM25), which is also expressed in a cytoplasmic form (pM25l), showed similar MLAs in the AC. Bioinformatics tools for predicting biomolecular condensates showed that four of the five proteins had a high propensity for liquid-liquid phase separation (LLPS), suggesting that LLPS may be a mechanism for compartmentalization within RC and AC. Examination of the physical properties of MLAs formed during the early phase of infection by 1,6-hexanediol treatment in vivo revealed liquid-like properties of pE1 MLAs and more solid-like properties of pM25 MLAs, indicating heterogeneity of mechanisms in the formation of virus-induced MLAs. Analysis of the five viral proteins and replicated viral DNA shows that the maturation sequence of RC and AC is not completed in many cells, suggesting that virus production and release is carried out by a rather limited number of cells. This study thus lays the groundwork for further investigation of the replication cycle of beta-herpesviruses, and the results should be incorporated into plans for high-throughput and single-cell analytic approaches.
MeSH term(s)	Animals ; Mice ; Muromegalovirus/metabolism ; DNA, Viral/genetics ; Cell Nucleus/metabolism ; Viral Proteins/genetics ; Viral Proteins/metabolism ; Viruses/metabolism ; Cytomegalovirus Infections
Chemical Substances	DNA, Viral ; Viral Proteins
Language	English
Publishing date	2023-03-16
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v15030766
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Article ; Online: Cytomegalovirus Infection: Mouse Model.

Brizić, Ilija / Lisnić, Berislav / Brune, Wolfram / Hengel, Hartmut / Jonjić, Stipan

Current protocols in immunology

2018 Volume 122, Issue 1, Page(s) e51

Abstract: This unit describes procedures for infecting newborn and adult mice with murine cytomegalovirus (MCMV). Methods are included for propagating MCMV in cell cultures and for preparing a more virulent form of MCMV from salivary glands of infected mice. A ... ...

Abstract	This unit describes procedures for infecting newborn and adult mice with murine cytomegalovirus (MCMV). Methods are included for propagating MCMV in cell cultures and for preparing a more virulent form of MCMV from salivary glands of infected mice. A plaque assay is provided for determining MCMV titers of infected tissues or virus stocks. Also, a method is described for preparing the murine embryonic fibroblasts used for propagating MCMV and for the plaque assay. © 2018 by John Wiley & Sons, Inc.
Language	English
Publishing date	2018-07-25
Publishing country	United States
Document type	Journal Article
ZDB-ID	2179059-0
ISSN	1934-368X ; 1934-3671
ISSN (online)	1934-368X
ISSN	1934-3671
DOI	10.1002/cpim.51
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Article: Distinguishing Natural Infections of the Bovine Mammary Gland by

Rešetar Maslov, Dina / Thomas, Funmilola Clara / Beletić, Anđelo / Kuleš, Josipa / Rubić, Ivana / Benić, Miroslav / Bačić, Goran / Maćešić, Nino / Eraghi, Vida / Farkaš, Vladimir / Lenac Roviš, Tihana / Lisnić, Berislav / Žubčić, Damir / Potočnjak, Dalibor / Mrljak, Vladimir

Animals : an open access journal from MDPI

2023 Volume 13, Issue 11

Abstract: Bovine mastitis is the most frequent disease on dairy farms, which leads to a decrease in the health welfare of the animals and great economic losses. This study was aimed at determining the quantitative variations in the milk proteome caused by natural ... ...

Abstract	Bovine mastitis is the most frequent disease on dairy farms, which leads to a decrease in the health welfare of the animals and great economic losses. This study was aimed at determining the quantitative variations in the milk proteome caused by natural infection by
Language	English
Publishing date	2023-05-31
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2606558-7
ISSN	2076-2615
ISSN	2076-2615
DOI	10.3390/ani13111829
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Article ; Online: Perinatal murine cytomegalovirus infection reshapes the transcriptional profile and functionality of NK cells.

Rožmanić, Carmen / Lisnić, Berislav / Pribanić Matešić, Marina / Mihalić, Andrea / Hiršl, Lea / Park, Eugene / Lesac Brizić, Ana / Indenbirken, Daniela / Viduka, Ina / Šantić, Marina / Adler, Barbara / Yokoyama, Wayne M / Krmpotić, Astrid / Juranić Lisnić, Vanda / Jonjić, Stipan / Brizić, Ilija

Nature communications

2023 Volume 14, Issue 1, Page(s) 6412

Abstract: Infections in early life can elicit substantially different immune responses and pathogenesis than infections in adulthood. Here, we investigate the consequences of murine cytomegalovirus infection in newborn mice on NK cells. We show that infection ... ...

Abstract	Infections in early life can elicit substantially different immune responses and pathogenesis than infections in adulthood. Here, we investigate the consequences of murine cytomegalovirus infection in newborn mice on NK cells. We show that infection severely compromised NK cell maturation and functionality in newborns. This effect was not due to compromised virus control. Inflammatory responses to infection dysregulated the expression of major transcription factors governing NK cell fate, such as Eomes, resulting in impaired NK cell function. Most prominently, NK cells from perinatally infected mice have a diminished ability to produce IFN-γ due to the downregulation of long non-coding RNA Ifng-as1 expression. Moreover, the bone marrow's capacity to efficiently generate new NK cells is reduced, explaining the prolonged negative effects of perinatal infection on NK cells. This study demonstrates that viral infections in early life can profoundly impact NK cell biology, including long-lasting impairment in NK cell functionality.
MeSH term(s)	Mice ; Animals ; Killer Cells, Natural ; Cytomegalovirus Infections/genetics ; Muromegalovirus
Language	English
Publishing date	2023-10-12
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-023-42182-w
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Article ; Online: NK cell interplay with cytomegaloviruses.

Lisnić, Berislav / Lisnić, Vanda Juranić / Jonjić, Stipan

Current opinion in virology

2015 Volume 15, Page(s) 9–18

Abstract: NK cells play an important role in the control of viral infections. Cytomegaloviruses have played a big part in the accumulation of current knowledge describing how NK cells recognize and eliminate infected cells. The interference with these functions at ...

Abstract	NK cells play an important role in the control of viral infections. Cytomegaloviruses have played a big part in the accumulation of current knowledge describing how NK cells recognize and eliminate infected cells. The interference with these functions at multiple levels may not only play a role in the control of primary infections or reactivations but can also impact other arms of the immune system and leave a long-lasting stable imprint on the NK cell population. These imprints may, in turn, modify how we respond to other infections. Understanding these processes will allow us to design better diagnostic approaches and new treatment options through manipulation of our immune responses and the viruses themselves.
MeSH term(s)	Animals ; Cytomegalovirus/immunology ; Cytomegalovirus Infections/immunology ; Cytomegalovirus Infections/virology ; Humans ; Immunity, Innate ; Killer Cells, Natural/immunology ; Killer Cells, Natural/virology ; Mice
Language	English
Publishing date	2015-12
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2611378-8
ISSN	1879-6265 ; 1879-6257
ISSN (online)	1879-6265
ISSN	1879-6257
DOI	10.1016/j.coviro.2015.07.001
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Article: The Virus-Induced Upregulation of the miR-183/96/182 Cluster and the FoxO Family Protein Members Are Not Required for Efficient Replication of HSV-1

Zubković, Andreja / Žarak, Ines / Ratkaj, Ivana / Rokić, Filip / Jekić, Maja / Pribanić Matešić, Marina / Lebrón, Ricardo / Gómez-Martín, Cristina / Lisnić, Berislav / Lisnić, Vanda Juranić / Jonjić, Stipan / Pan, Dongli / Vugrek, Oliver / Hackenberg, Michael / Jurak, Igor

Viruses. 2022 July 28, v. 14, no. 8

2022

Abstract: Herpes simplex virus 1 (HSV-1) expresses a large number of miRNAs, and their function is still not completely understood. In addition, HSV-1 has been found to deregulate host miRNAs, which adds to the complexity of the regulation of efficient virus ... ...

Abstract	Herpes simplex virus 1 (HSV-1) expresses a large number of miRNAs, and their function is still not completely understood. In addition, HSV-1 has been found to deregulate host miRNAs, which adds to the complexity of the regulation of efficient virus replication. In this study, we comprehensively addressed the deregulation of host miRNAs by massive-parallel sequencing. We found that only miRNAs expressed from a single cluster, miR-183/96/182, are reproducibly deregulated during productive infection. These miRNAs are predicted to regulate a great number of potential targets involved in different cellular processes and have only 33 shared targets. Among these, members of the FoxO family of proteins were identified as potential targets for all three miRNAs. However, our study shows that the upregulated miRNAs do not affect the expression of FoxO proteins, moreover, these proteins were upregulated in HSV-1 infection. Furthermore, we show that the individual FoxO proteins are not required for efficient HSV-1 replication. Taken together, our results indicate a complex and redundant response of infected cells to the virus infection that is efficiently inhibited by the virus.
Keywords	Human alphaherpesvirus 1 ; microRNA ; virus replication ; viruses
Language	English
Dates of publication	2022-0728
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2516098-9
ISSN	1999-4915
ISSN	1999-4915
DOI	10.3390/v14081661
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Size-dependent antirecombinogenic effect of short spacers on palindrome recombinogenicity.

Svetec Miklenić, Marina / Gatalica, Nikolina / Matanović, Angela / Žunar, Bojan / Štafa, Anamarija / Lisnić, Berislav / Svetec, Ivan Krešimir

DNA repair

2020 Volume 90, Page(s) 102848

Abstract: Palindromic sequences in DNA can instigate genetic recombination and genome instability, which can result in devastating conditions such as the Emmanuel syndrome. Palindrome recombinogenicity increases with its size and sequence similarity between ... ...

Abstract	Palindromic sequences in DNA can instigate genetic recombination and genome instability, which can result in devastating conditions such as the Emmanuel syndrome. Palindrome recombinogenicity increases with its size and sequence similarity between palindrome arms, while quasipalindromes with long spacers are less recombinogenic. However, the minimal spacer length, which could reduce or abolish palindrome recombinogenicity in the eukaryotic genome, was never determined. Therefore, we constructed a series of palindromes containing spacers of lengths ranging from 0 (perfect palindrome) to 10 bp and tested their recombinogenicity in yeast Saccharomyces cerevisiae. We found that a 7 bp spacer significantly reduces 126 bp palindrome recombinogenicity, while a 10 bp spacer completely stabilizes palindromes up to 150 bp long. Additionally, we showed that palindrome stimulated recombination rate is not dependent on Mus81 and Yen1 endonucleases. We also compared the recombinogenicity of a perfect 126 bp palindrome and a corresponding quasipalindrome consisting of the same palindrome arms with a stabilising 10 bp spacer in sgs1Δ and rad27Δ backgrounds, since both Sgs1 helicase and Rad27 endonuclease are implicated in preventing hairpin formation at palindromic sequences during replication.
MeSH term(s)	Base Sequence ; DNA, Fungal/metabolism ; Flap Endonucleases/metabolism ; Inverted Repeat Sequences ; RecQ Helicases/metabolism ; Recombination, Genetic ; Saccharomyces cerevisiae/enzymology ; Saccharomyces cerevisiae/genetics ; Saccharomyces cerevisiae Proteins/metabolism
Chemical Substances	DNA, Fungal ; Saccharomyces cerevisiae Proteins ; Flap Endonucleases (EC 3.1.-) ; RAD27 protein, S cerevisiae (EC 3.1.11.5) ; SGS1 protein, S cerevisiae (EC 3.6.1.-) ; RecQ Helicases (EC 3.6.4.12)
Language	English
Publishing date	2020-05-03
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2071608-4
ISSN	1568-7856 ; 1568-7864
ISSN (online)	1568-7856
ISSN	1568-7864
DOI	10.1016/j.dnarep.2020.102848
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