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Article ; Online: Relationship of human cytomegalovirus-infected endothelial cells and circulating leukocytes in the pathogenesis of disseminated human cytomegalovirus infection: A narrative review.

Gerna, Giuseppe / Lilleri, Daniele / Fornara, Chiara / d'Angelo, Piera / Baldanti, Fausto

2024 Volume 34, Issue 1, Page(s) e2496

Abstract: Among the leucocyte subpopulations circulating in peripheral blood of immune-compromised patients with disseminated Human cytomegalovirus (HCMV) infection, polymorphonuclear leuckocytes (PMNL) and M/M may carry infectious virus. While only in PMNL early ... ...

Abstract	Among the leucocyte subpopulations circulating in peripheral blood of immune-compromised patients with disseminated Human cytomegalovirus (HCMV) infection, polymorphonuclear leuckocytes (PMNL) and M/M may carry infectious virus. While only in PMNL early HCMV replicative events do occur, monocytes are susceptible to complete virus replication when they enter human organs, where as macrophages become a site of active complete virus replication. In vivo leucocytes and endothelial cells interact continuously, as suggested by several in vitro experimental findings showing the bidirectional HCMV transmission from leucocytes to and from endothelial cells with the critical aid of adhesion molecules. Recently, the neutralising antibody response in sera from subjects with primary HCMV infection was reported to be much higher and earlier than in human embryonic lung fibroblasts (HELF) cells when measured in endothelial cells and epithelial cells, where virus entry is mediated mostly by the pentamer complex gH/gL/pUL128/pUL130/pUL131, whereas it was much lower and delayed when determined in HELF, where virus entry is mediated mostly by the trimer complex gH/gL/gO. Thus, these results suggested that products of UL128L were the molecules primary responsible for the differential neutralising antibody response. This conclusion was confirmed by a series of polyclonal and monoclonal antibodies directed to the components of pUL128L. Very recently, based on two sets of experiments including inhibition and immunoblotting assays, the pentamer complex/trimer complex ratio has been finally identified as the main factor of the neutralising antibody response. This ratio may change with the virus suspension producer and target cell system as well as number of cell culture passages.
MeSH term(s)	Humans ; Cytomegalovirus ; Endothelial Cells ; Viral Envelope Proteins ; Cytomegalovirus Infections ; Antibodies, Neutralizing ; Virus Internalization ; Leukocytes
Chemical Substances	Viral Envelope Proteins ; Antibodies, Neutralizing
Language	English
Publishing date	2024-01-29
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	1086043-5
ISSN	1099-1654 ; 1052-9276
ISSN (online)	1099-1654
ISSN	1052-9276
DOI	10.1002/rmv.2496
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Article: Human Cytomegalovirus Congenital (cCMV) Infection Following Primary and Nonprimary Maternal Infection: Perspectives of Prevention through Vaccine Development.

Gerna, Giuseppe / Lilleri, Daniele

Vaccines

2020 Volume 8, Issue 2

Abstract: Congenital cytomegalovirus (cCMV) might occur as a result of the human cytomegalovirus (HCMV) primary (PI) or nonprimary infection (NPI) in pregnant women. Immune correlates of protection against cCMV have been partly identified only for PI. Following ... ...

Abstract	Congenital cytomegalovirus (cCMV) might occur as a result of the human cytomegalovirus (HCMV) primary (PI) or nonprimary infection (NPI) in pregnant women. Immune correlates of protection against cCMV have been partly identified only for PI. Following either PI or NPI, HCMV strains undergo latency. From a diagnostic standpoint, while the serological criteria for the diagnosis of PI are well-established, those for the diagnosis of NPI are still incomplete. Thus far, a recombinant gB subunit vaccine has provided the best results in terms of partial protection. This partial efficacy was hypothetically attributed to the post-fusion instead of the pre-fusion conformation of the gB present in the vaccine. Future efforts should be addressed to verify whether a new recombinant gB pre-fusion vaccine would provide better results in terms of prevention of both PI and NPI. It is still a matter of debate whether human hyperimmune globulin are able to protect from HCMV vertical transmission. In conclusion, the development of an HCMV vaccine that would prevent a significant portion of PI would be a major step forward in the development of a vaccine for both PI and NPI.
Language	English
Publishing date	2020-04-23
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2703319-3
ISSN	2076-393X
ISSN	2076-393X
DOI	10.3390/vaccines8020194
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Article ; Online: Brief molecular diagnostic criteria for human cytomegalovirus infection/disease.

Gerna, Giuseppe / Baldanti, Fausto

Expert review of molecular diagnostics

2019 Volume 19, Issue 9, Page(s) 773–775

MeSH term(s)	Cytomegalovirus/genetics ; Cytomegalovirus Infections/diagnosis ; Humans ; Molecular Diagnostic Techniques/methods ; Pathology, Molecular/methods ; Transplants/virology
Language	English
Publishing date	2019-08-22
Publishing country	England
Document type	Editorial
ZDB-ID	2112530-2
ISSN	1744-8352 ; 1473-7159
ISSN (online)	1744-8352
ISSN	1473-7159
DOI	10.1080/14737159.2019.1657010
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Article ; Online: Fibroblast, Epithelial and Endothelial Cell-Derived Human Cytomegalovirus Strains Display Distinct Neutralizing Antibody Responses and Varying Levels of gH/gL Complexes.

Fornara, Chiara / Schultz, Eric / Lilleri, Daniele / Baldanti, Fausto / Ryckman, Brent / Gerna, Giuseppe

International journal of molecular sciences

2023 Volume 24, Issue 5

Abstract: In sequential sera from pregnant women with HCMV primary infection (PI), the serum neutralizing activity is higher against virions produced in epithelial and endothelial cells than in fibroblasts. Immunoblotting shows that the pentamer complex/trimer ... ...

Abstract	In sequential sera from pregnant women with HCMV primary infection (PI), the serum neutralizing activity is higher against virions produced in epithelial and endothelial cells than in fibroblasts. Immunoblotting shows that the pentamer complex/trimer complex (PC/TC) ratio varies according to the producer cell culture type used for the virus preparation to be employed in the neutralizing antibody (NAb) assay, and is lower in fibroblasts and higher in epithelial, and especially endothelial cells. The blocking activity of TC- and PC-specific inhibitors varies according to the PC/TC ratio of virus preparations. The rapid reversion of the virus phenotype following its back passage to the original cell culture (fibroblasts) potentially argues in favor of a producer cell effect on virus phenotype. However, the role of genetic factors cannot be overlooked. In addition to the producer cell type, the PC/TC ratio may differ in single HCMV strains. In conclusion, the NAb activity not only varies with different HCMV strains, but is a dynamic parameter changing according to virus strain, type of target and producer cells, and number of cell culture passages. These findings may have some important implications for the development of both therapeutic antibodies and subunit vaccines.
MeSH term(s)	Humans ; Female ; Pregnancy ; Cytomegalovirus ; Endothelial Cells/metabolism ; Viral Envelope Proteins/genetics ; Membrane Glycoproteins/metabolism ; Cytomegalovirus Infections ; Antibodies, Neutralizing ; Fibroblasts/metabolism
Chemical Substances	Viral Envelope Proteins ; Membrane Glycoproteins ; Antibodies, Neutralizing
Language	English
Publishing date	2023-02-23
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms24054417
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Congenital Human Cytomegalovirus Infection: A Narrative Review of Maternal Immune Response and Diagnosis in View of the Development of a Vaccine and Prevention of Primary and Non-Primary Infections in Pregnancy.

Gerna, Giuseppe / Fornara, Chiara / Furione, Milena / Lilleri, Daniele

Microorganisms

2021 Volume 9, Issue 8

Abstract: Congenital cytomegalovirus infection (cCMV) may affect about 1% of all newborns all over the world as a result of either a primary or recurrent human cytomegalovirus (HCMV) infection. While about 90% of infants affected by cCMV are asymptomatic at birth, ...

Abstract	Congenital cytomegalovirus infection (cCMV) may affect about 1% of all newborns all over the world as a result of either a primary or recurrent human cytomegalovirus (HCMV) infection. While about 90% of infants affected by cCMV are asymptomatic at birth, the remaining 10% are symptomatic often with neurodevelopmental impairment and sensorineural hearing loss. In view of identifying the best approach to vaccine prevention of cCMV, this review will examine the most important steps made in the study of the immune response to, and diagnosis of, HCMV infection. The maternal immune response and immune correlates of protection are being partially identified with a partial contribution given by our laboratory. The diagnosis of primary infection is often difficult to achieve in the first three months of pregnancy, which is the time primarily involved in virus transmission to the fetus in association with the most severe symptoms and
Language	English
Publishing date	2021-08-16
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2720891-6
ISSN	2076-2607
ISSN	2076-2607
DOI	10.3390/microorganisms9081749
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Article: Human cytomegalovirus (HCMV) infection/re-infection: development of a protective HCMV vaccine.

Gerna, Giuseppe / Lilleri, Daniele

The new microbiologica

2019 Volume 42, Issue 1, Page(s) 1–20

Abstract: In recent years, one of the main objectives in the field of medical virology has been the development of a human cytomegalovirus (HCMV) vaccine that can prevent congenital HCMV infection in the offspring of pregnant women as well as systemic and end- ... ...

Abstract	In recent years, one of the main objectives in the field of medical virology has been the development of a human cytomegalovirus (HCMV) vaccine that can prevent congenital HCMV infection in the offspring of pregnant women as well as systemic and end-organ disease in immunocompromised (AIDS and transplanted) patients. Major obstacles to the development of an efficacious HCMV vaccine are lack of protection provided by immune memory cells against HCMV re-activation (replication relapse of a latent strain following primary infection) and HCMV re-infection (infection of a seropositive individual by a new virus strain). Thus, while initial efforts were directed at the development of a vaccine for the prevention of primary infection, in the last decade the primary vaccine development endpoint was the prevention of primary HCMV infection, as well as HCMV re-activation and re-infection. Along this line of research, both HCMV live (including Towne, AD169 and its derivatives, Towne/Toledo chimeras, Viral-Vectored vaccines, and Virus Replicon Particles) and non-living vaccines (including the recombinant gB subunit, DNA- and RNA-based vaccines, Virus-like particles, Dense bodies, Peptide vaccines, and the Pentamer Complex) have been developed. To date, Phase I, II, and III clinical trials have been variably conducted in humans, and experimental inoculation in different animal models has been performed with different vaccine formulations. Notwithstanding the variable research conditions, clinical and experimental results achieved thus far predict that the ideal HCMV vaccine should be able to elicit both robust humoral (both neutralizing and binding) and HCMV-specific CD4+ and CD8+ T-cell responses. This vaccine should hypothetically contain: 1) gB, inducing both humoral and T-cell responses; 2) the pentameric complex (PC), inducing the most potent neutralizing antibody response; 3) pp65, inducing the most potent HCMV-specific T-cell response. Although the protective role of cell-mediated immunity has been repeatedly documented, while the protective effect of (mostly neutralizing) antibodies remains partly to be documented, a vaccine stimulating both arms of the immune response would likely confer the highest level of protection against HCMV infection/disease in both HCMV-seronegative and -seropositive individuals.
MeSH term(s)	Animals ; Cytomegalovirus/immunology ; Cytomegalovirus Infections/prevention & control ; Cytomegalovirus Vaccines ; Female ; Humans ; Pregnancy
Chemical Substances	Cytomegalovirus Vaccines
Language	English
Publishing date	2019-01-21
Publishing country	Italy
Document type	Journal Article ; Review
ZDB-ID	756168-4
ISSN	1121-7138 ; 0391-5352
ISSN	1121-7138 ; 0391-5352
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Zs.B 2267: Show issues

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Article: Congenital Human Cytomegalovirus Infection: A Narrative Review of Maternal Immune Response and Diagnosis in View of the Development of a Vaccine and Prevention of Primary and Non-Primary Infections in Pregnancy

Gerna, Giuseppe / Fornara, Chiara / Furione, Milena / Lilleri, Daniele

Microorganisms. 2021 Aug. 16, v. 9, no. 8

2021

Abstract	Congenital cytomegalovirus infection (cCMV) may affect about 1% of all newborns all over the world as a result of either a primary or recurrent human cytomegalovirus (HCMV) infection. While about 90% of infants affected by cCMV are asymptomatic at birth, the remaining 10% are symptomatic often with neurodevelopmental impairment and sensorineural hearing loss. In view of identifying the best approach to vaccine prevention of cCMV, this review will examine the most important steps made in the study of the immune response to, and diagnosis of, HCMV infection. The maternal immune response and immune correlates of protection are being partially identified with a partial contribution given by our laboratory. The diagnosis of primary infection is often difficult to achieve in the first three months of pregnancy, which is the time primarily involved in virus transmission to the fetus in association with the most severe symptoms and sequelae. Prevention of cCMV is anticipated by prevention of primary infection in early pregnancy by means of different measures, such as (i) behavioral-educational measures, (ii) immunoglobulin administration, (iii) antiviral treatment with valaciclovir. However, the most promising approach to cCMV prevention appears to be the development of a non-living vaccine, including at least three viral antigens: gB, pentamer complex gHgLpUL128L, and pp65, which have been shown to be able to stimulate both the humoral and the cellular arms of the maternal immune response. Primary HCMV infection may be managed in pregnancy by counseling of the couples involved by a team of specialists that includes virologists, obstetricians, infectivologists and neonatologists.
Keywords	Cytomegalovirus infections ; Human betaherpesvirus 5 ; complications (disease) ; fetus ; immune response ; immunoglobulins ; pregnancy ; vaccines ; virus transmission
Language	English
Dates of publication	2021-0816
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2720891-6
ISSN	2076-2607
ISSN	2076-2607
DOI	10.3390/microorganisms9081749
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Human Cytomegalovirus Congenital (cCMV) Infection Following Primary and Nonprimary Maternal Infection

Giuseppe Gerna / Daniele Lilleri

Vaccines, Vol 8, Iss 194, p

Perspectives of Prevention through Vaccine Development

2020 Volume 194

Abstract	Congenital cytomegalovirus (cCMV) might occur as a result of the human cytomegalovirus (HCMV) primary (PI) or nonprimary infection (NPI) in pregnant women. Immune correlates of protection against cCMV have been partly identified only for PI. Following either PI or NPI, HCMV strains undergo latency. From a diagnostic standpoint, while the serological criteria for the diagnosis of PI are well-established, those for the diagnosis of NPI are still incomplete. Thus far, a recombinant gB subunit vaccine has provided the best results in terms of partial protection. This partial efficacy was hypothetically attributed to the post-fusion instead of the pre-fusion conformation of the gB present in the vaccine. Future efforts should be addressed to verify whether a new recombinant gB pre-fusion vaccine would provide better results in terms of prevention of both PI and NPI. It is still a matter of debate whether human hyperimmune globulin are able to protect from HCMV vertical transmission. In conclusion, the development of an HCMV vaccine that would prevent a significant portion of PI would be a major step forward in the development of a vaccine for both PI and NPI.
Keywords	congenital cytomegalovirus ; primary infection ; non-primary infection ; gB ; neutralizing antibodies ; pentameric complex ; Medicine ; R
Subject code	570
Language	English
Publishing date	2020-04-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Endothelial cells and CMV dissemination.

Gerna, Giuseppe

Future microbiology

2012 Volume 7, Issue 4, Page(s) 441–444

Abstract: Using the murine CMV animal model and the well-established model of Cre-lox-P-mediated green-fluorescence tagging of endothelial cell (EC)-derived mouse CMV to quantify the role of infected ECs in transplantation-associated CMV dissemination (in mice ... ...

Abstract	Using the murine CMV animal model and the well-established model of Cre-lox-P-mediated green-fluorescence tagging of endothelial cell (EC)-derived mouse CMV to quantify the role of infected ECs in transplantation-associated CMV dissemination (in mice expressing Cre recombinase under the control of either the Tie2 or the Tek promoter selectively expressed in vascular EC-Tie-Cre and Tek-Cre mice), it was shown that EC-derived virus contributed to 50% of the total viral load during primary infection, and there was no preference for dissemination of EC-derived viruses over viruses produced by other cell types. In addition, during secondary viremia, there was only a negligible contribution of EC-derived virus to dissemination to other organs. These results are novel in the methodology employed and are somewhat interesting. However, the data are limited to the mouse model with a short-term follow-up, and the immunodeficient host has not yet been studied. In humans, these conclusions must be taken with caution. First, in primary infection occurring through natural routes, epithelial cells are infected first, then ECs, unless primary infection occurs through blood transfusion, in which case endothelial vascular cells may become infected first. In both cases, the virus transport occurs through the intervention of leukocytes, namely monocytes and polymorphonuclear leukocytes. As monocytes differentiate to macrophages, they become highly susceptible to human CMV replication inside organ tissues, while polymorphonuclear leukocytes are active in virus capturing from infected endothelial vascular cells and transporting to distant sites.
Language	English
Publishing date	2012-04
Publishing country	England
Document type	Comment ; Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1746-0921
ISSN (online)	1746-0921
DOI	10.2217/fmb.12.12
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Article ; Online: Fibroblast, Epithelial and Endothelial Cell-Derived Human Cytomegalovirus Strains Display Distinct Neutralizing Antibody Responses and Varying Levels of gH/gL Complexes

Chiara Fornara / Eric Schultz / Daniele Lilleri / Fausto Baldanti / Brent Ryckman / Giuseppe Gerna

International Journal of Molecular Sciences, Vol 24, Iss 4417, p

2023 Volume 4417

Abstract	In sequential sera from pregnant women with HCMV primary infection (PI), the serum neutralizing activity is higher against virions produced in epithelial and endothelial cells than in fibroblasts. Immunoblotting shows that the pentamer complex/trimer complex (PC/TC) ratio varies according to the producer cell culture type used for the virus preparation to be employed in the neutralizing antibody (NAb) assay, and is lower in fibroblasts and higher in epithelial, and especially endothelial cells. The blocking activity of TC- and PC-specific inhibitors varies according to the PC/TC ratio of virus preparations. The rapid reversion of the virus phenotype following its back passage to the original cell culture (fibroblasts) potentially argues in favor of a producer cell effect on virus phenotype. However, the role of genetic factors cannot be overlooked. In addition to the producer cell type, the PC/TC ratio may differ in single HCMV strains. In conclusion, the NAb activity not only varies with different HCMV strains, but is a dynamic parameter changing according to virus strain, type of target and producer cells, and number of cell culture passages. These findings may have some important implications for the development of both therapeutic antibodies and subunit vaccines.
Keywords	human cytomegalovirus ; primary infection ; neutralizing antibodies ; trimer ; pentamer ; receptors ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
Subject code	570
Language	English
Publishing date	2023-02-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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