Human CMV‐IGIV (CytoGam®) neutralizes human cytomegalovirus (HCMV) infectivity and prevents intracellular signal transduction after HCMV exposure

Andreoni, Kenneth A.; Wang, X.; Huong, Shu-Mei; Huang, Eng‐Shang

doi:10.1034/j.1399-3062.2001.00005.x

Cited by 8 publications

(8 citation statements)

References 11 publications

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“…Although CMV-IC generation was not verified experimentally, high concentrations of CMV antigens and CMVhyperimmune globulin should inevitably lead to IC formation. 17 Wild-type CMV infection may impair DC phenotype/function 18 ; however, CMV-IC-loaded Mo/DCs expressed potent T cellstimulatory phenotype (Figure 1). …”

Section: Cmv-ic-loaded Cb Dcs Exhibit a Potent T Cell-stimulatory Phementioning

confidence: 99%

In vitro priming and expansion of cytomegalovirus-specific Th1 and Tc1 T cells from naive cord blood lymphocytes

Park

Marti

Kurtzberg

et al. 2006

Blood

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Adoptive transfer of CMV-specific cytotoxic T cells (CTLs) expanded in vitro from memory donor T cells can reduce the incidence of CMV disease in allogeneic transplant recipients. However, this approach has been unavailable in the cord blood (CB) transplantation setting because CB T cells are antigen naive and biased toward Th2/Tc2 function. We developed a protocol to in vitro prime and expand CMV-specific CTLs from CB. T cells were primed with cytokines to trigger skewing toward Th1/Tc1 lineage before encountering monocyte and CD34 ؉ progenitor-derived dendritic cells loaded with CMV antigen and its immune complex. CMV-pulsed cultures expanded significantly more over 4 to 6 weeks than CMV cultures despite identical cytokine milieu. T cells isolated from CMV ؉ cultures showed a preferential expansion of CD45RA ؊ /RO ؉ /CD27 ؉ T cells compared to CMV ؊ cultures. CMV-specific IFN-␥-and TNF-␣-producing CD4 ؉ (Th1) and CD8 ؉ (Tc1) T cells were enriched after 3 to 4 weeks and CMV-specific cytotoxicity developed 1 to 2 weeks later. Although CMV-specific CTL generation is feasible in seropositive donors, it rarely succeeds in CMV-seronegative individuals. 7 Antiviral CTLs have been unavailable for umbilical cord blood transplant (UCBT) recipients because cord blood (CB), like seronegative donors, contains antigen-inexperienced T cells. Additionally, compared to CD45RA ϩ /CD28 ϩ 'naive' adult T cells, CB T cells are defective in the inducible expression of Th1/Tc1 cytokines, primarily IFN-␥, as a result of Th2/Tc2 skewing imprinted by placental factors (for a review, see Marchant and Goldman 8 ). Independently, diminished IL-12 expression by neonatal DCs impairs Th1 immunity. 9,10 We now report that CMV-specific CTLs can be generated from CB along the Th1/Tc1 pathway. Study design DC generation and antigen loadingFresh mononuclear cells (MNCs) from discarded CB units obtained with Institutional Review Board approval were cultured in RPMI 1640 plus 10% pooled human serum (PHS; NABI, Miami, FL). CD34 ϩ progenitors were selected from nonadherent cells 11 using CD34-PE (Becton Dickinson, San Jose, CA) and anti-PE microbeads (Miltenyi Biotech, Auburn, CA) over a MiniMACS column per the manufacturer's instruction. CD34 Ϫ cells were frozen. Immature Mo/DCs developed from adherent monocytes in RPMI 1640 plus 1% PHS, GM-CSF, and IL-4 over 5 to 6 days. 11 Unused MNCs were frozen and subsequently thawed weekly until exhausted to generate immature Mo/DCs as described except for the adherence media, LGM-3 plus 10% PHS (Clonetics, San Diego, CA). On "day -2" DCs were pulsed at 37°C with inactivated AD169 CMV lysate (5 g/mL, Biodesign, Carmel, NY). One hour later 1 mg/mL CMV hyperimmune globulin (Medimmune, Gaithersburg, MD) was added for 6 to 8 hours, followed by maturation in TNF-␣ plus PGE 2 11,12 for 24 to 36 hours. Once Mo/DCs were exhausted CD34 ϩ -derived DCs were generated in AIM V (Gibco, Grand Island, NY) plus 3% PHS, 20 ng/mL SCF, 100 ng/mL Flt-3L (both from Amgen, Thousand Oaks, CA), GM-CSF, and IL-4. After 12 to 14 days, ...

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Section: Cmv-ic-loaded Cb Dcs Exhibit a Potent T Cell-stimulatory Phementioning

confidence: 99%

In vitro priming and expansion of cytomegalovirus-specific Th1 and Tc1 T cells from naive cord blood lymphocytes

Park

Marti

Kurtzberg

et al. 2006

Blood

View full text Add to dashboard Cite

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“…Moreover, administration of Ig preparations has been proposed repeatedly for the prevention and therapy of HCMV infection and disease in both immunocompetent and immunocompromised patients (Bass et al, 1993;Messori et al, 1994;Andreoni et al, 2001;Nigro et al, 2005); neutralizing human mAbs have been developed for the same purpose (Boeckh et al, 2001). Again, our study suggests that a complete evaluation of the protective activity of Ig preparations/human mAbs must include an assay in HUVECs or ARPE-19 cells, and that anti-pUL131A, -pUL130 and -pUL128 human mAbs may be a valuable component in the formulation of such immunodrugs.…”

mentioning

confidence: 99%

Human cytomegalovirus serum neutralizing antibodies block virus infection of endothelial/epithelial cells, but not fibroblasts, early during primary infection

Gerna

Sarasini

Patrone

et al. 2008

Journal of General Virology

169

172

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A panel of human sera exhibited a ¢128-fold higher neutralizing potency against a human cytomegalovirus (HCMV) clinical isolate propagated and tested in endothelial (or epithelial) cells than against the same virus infecting human fibroblasts. In a group of 18 primary infections, the reverse geometric mean titre was in the range of 10-15 in human fibroblasts within the first 3 months after the onset of infection, whereas the endothelial cell infection-neutralizing activity was already present within the first 10 days, reaching median levels of 122, 320 and 545 at respectively 30, 60 and 90 days after onset, then declining slowly. This difference was also confirmed in the majority of reactivated and remote HCMV infections, as well as in a hyperimmune globulin preparation. The antibody response to HCMV pUL131A, pUL130 and pUL128 locus products, which are required for endothelial/epithelial cell infection, provided a potential molecular basis for such a differential neutralizing activity. In addition, monoclonal/monospecific antibodies raised against the pUL131A, pUL130 and pUL128 proteins were found to display an inhibitory activity on HCMV plaque formation and HCMV leukocyte transfer from HCMV-infected cells. Hence, conventional determination of the neutralizing activity of human sera in fibroblasts is misleading. Antibodies to pUL131A, pUL130 and pUL128 appear to display a major HCMV-neutralizing and dissemination-inhibiting activity.

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“…Cytogam is used as a prophylaxis to treat CMV disease. The addition of Cytogam to HCMV has been previously demonstrated to neutralize virus and prevent viral entry [39]. Following treatment with Cytogam, we did not observe GFP-positive fibroblasts at 4 dpi (Fig.…”

Section: Resultsmentioning

confidence: 60%

“…In these studies, the addition of purified HCMV or UV-inactivated purified virus to the tissues resulted in a significant drop in the contractile force (Fig. 3A) [39]. Tissue mechanical disruption was observed for both the fibroblast-tropic HCMV strain, AD169, as well as the clinical strain, TB40/E (Fig.…”

Section: Discussionmentioning

confidence: 86%

A Method for Quantifying Mechanical Properties of Tissue following Viral Infection

et al. 2012

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Viral infection and replication involves the reorganization of the actin network within the host cell. Actin plays a central role in the mechanical properties of cells. We have demonstrated a method to quantify changes in mechanical properties of fabricated model three-dimensional (3D) connective tissue following viral infection. Using this method, we have characterized the impact of infection by the human herpesvirus, cytomegalovirus (HCMV). HCMV is a member of the herpesvirus family and infects a variety of cell types including fibroblasts. In the body, fibroblasts are necessary for maintaining connective tissue and function by creating mechanical force. Using this 3D connective tissue model, we observed that infection disrupted the cell’s ability to generate force and reduced the cumulative contractile force of the tissue. The addition of HCMV viral particles in the absence of both viral gene expression and DNA replication was sufficient to disrupt tissue function. We observed that alterations of the mechanical properties are, in part, due to a disruption of the underlying complex actin microfilament network established by the embedded fibroblasts. Finally, we were able to prevent HCMV-mediated disruption of tissue function by the addition of human immune globulin against HCMV. This study demonstrates a method to quantify the impact of viral infection on mechanical properties which are not evident using conventional cell culture systems.

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Human CMV‐IGIV (CytoGam®) neutralizes human cytomegalovirus (HCMV) infectivity and prevents intracellular signal transduction after HCMV exposure

Cited by 8 publications

References 11 publications

In vitro priming and expansion of cytomegalovirus-specific Th1 and Tc1 T cells from naive cord blood lymphocytes

In vitro priming and expansion of cytomegalovirus-specific Th1 and Tc1 T cells from naive cord blood lymphocytes

Human cytomegalovirus serum neutralizing antibodies block virus infection of endothelial/epithelial cells, but not fibroblasts, early during primary infection

A Method for Quantifying Mechanical Properties of Tissue following Viral Infection

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