Study of the Transmission of Cytomegalovirus (CMV) Infection From Mother to Foetus

Study Overview
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Brief Summary
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This study is designed to evaluate maternal virological and immunological parameters to determine their ability to predict congenital cytomegalovirus (CMV) infection. When a pregnant woman is infected with CMV, her immune system (which protects her from infection) is activated and the virus can be found in the woman's bodily fluids (blood, saliva, urine, vaginal secretions). The aim of this study is to find out if there is a link between either the pregnant woman's immune response or the presence of the virus in these bodily fluids and the child/foetus being infected with the virus.

Official Title
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Study of Maternal-foetal Cytomegalovirus (CMV) Transmission

Conditions
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Infections, Cytomegalovirus, Cytomegalovirus Infections

Intervention / Treatment
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* Procedure: Blood sample
* Procedure: Cord blood sample
* Procedure: Saliva swab
* Procedure: Urine sampling
* Procedure: Vaginal swab


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Subjects who the investigator believes that they can and will comply with the requirements of the protocol . A pregnant female, 18 years of age or older at the time of study enrolment. Women with confirmed primary CMV infection. Written informed consent obtained from the subject. Exclusion Criteria: Chronic administration of immunosuppressants or other immune-modifying drugs within six months prior to study entry. Concurrently participating in another clinical study, at any time during the study period, in which the subject has been or will be exposed to an investigational or a non-investigational pharmaceutical product. Previous vaccination against CMV infection. Any confirmed or suspected immunosuppressive or immunodeficient condition, based on medical history or physical examination Major congenital defects, serious chronic illness or organ transplantation. Administration of immunoglobulins and/or any blood products within the three months preceding study enrolment or during the pregnancy. Documented Human immunodeficiency virus (HIV)-positive subject. Gestational age of more than 34 weeks, as determined by foetal ultrasound.

Ages Eligible for Study
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Minimum Age: 18 Years

Sexes Eligible for Study
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Female

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Other
Allocation: Non-Randomized
Intervention Model: Single Group Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: CMV Mothers' Group<br>Pregnant subjects with confirmed primary CMV infection. | Procedure: Blood sample<br>* Blood sample at study entry, every two months during pregnancy, at pregnancy conclusion and one month after pregnancy conclusion.<br>Procedure: Cord blood sample<br>* Cord blood sample taken at the time of delivery.<br>Procedure: Saliva swab<br>* Saliva swab taken at study entry, every month during pregnancy, at pregnancy conclusion and one month after pregnancy conclusion.<br>Procedure: Urine sampling<br>* Approximately 10 mL of urine will be sampled at study entry, every month during pregnancy, at pregnancy conclusion and one month after pregnancy conclusion.<br>Procedure: Vaginal swab<br>* Vaginal swab taken at study entry, every month during pregnancy and one month after pregnancy conclusion.<br>|
| Experimental: CMV Newborns' Group<br>Offsprings of the CMV Mothers' Group, also tested for CMV infection, comprising infants that were live born. | Procedure: Blood sample<br>* Blood sample at study entry, every two months during pregnancy, at pregnancy conclusion and one month after pregnancy conclusion.<br>Procedure: Cord blood sample<br>* Cord blood sample taken at the time of delivery.<br>Procedure: Saliva swab<br>* Saliva swab taken at study entry, every month during pregnancy, at pregnancy conclusion and one month after pregnancy conclusion.<br>Procedure: Urine sampling<br>* Approximately 10 mL of urine will be sampled at study entry, every month during pregnancy, at pregnancy conclusion and one month after pregnancy conclusion.<br>Procedure: Vaginal swab<br>* Vaginal swab taken at study entry, every month during pregnancy and one month after pregnancy conclusion.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Number of Subjects With Any Cytomegalovirus (CMV) Congenital Infection | The CMV congenital infections were assessed in newborns and foetuses of subjects who had a confirmed primary CMV infection during pregnancy. | At Month 0 | 
| Number of Subjects With CMV Presence in the Urine | Evidence of infection in urine was assessed by culture or by Polymerase Chain Reaction (PCR). | Within 10 days post-delivery (Days 0-9) | 
| Number of Subjects With CMV Presence in the Amniotic Fluid | Evidence of infection in the amniotic fluid was assessed by culture or by Polymerase Chain Reaction (PCR). | Within 10 days post-delivery (Days 0-9) | 
| Evidence of CMV DNA or CMV Inclusions in Tissues of an Aborted or Stillborn Foetus |  | Within 10 days post-delivery (Days 0-9) | 
| Number of CMV DNA Copies in Saliva, Urine, Blood or Vaginal Secretions | The assessment focused on the presence of CMV DNA copies (by Quantitative Polymerase Chain Reaction [qPCR]) in saliva, urine, blood and vaginal secretions every month from study entry to, and including, pregnancy conclusion. | At Month 0 | 
| Number of CMV DNA Copies in Saliva, in Urine and in Blood or Vaginal Secretions | The assessment focused on the presence of CMV DNA copies (by Quantitative Polymerase Chain Reaction [qPCR]) in saliva, urine and blood every month from study entry to, and including, pregnancy conclusion. | At pregnancy conclusion (Day 0 to 5, Day 0 = day of delivery, stillbirth or termination) | 
| Descriptive Statistics of the Anti-CMV Immunoglobulin Type M (IgM) Status | Anti-CMV IgM status was assessed by Enzyme-Linked Immunosorbent Assay [ELISA], with respect to positive and negative subjects. Grey Zone = optical density zone within 20% of cut-off value. When an optical density is within this grey zone, sample testing is repeated to confirm the result. | At Month 0 | 
| Descriptive Statistics of the Anti-CMV IgM Status | Anti-CMV IgM status was assessed by Enzyme-Linked Immunosorbent Assay [ELISA], with respect to positive and negative subjects. Grey Zone = optical density zone within 20% of cut-off value. When an optical density is within this grey zone, sample testing is repeated to confirm the result. | At Month 2 | 
| Descriptive Statistics of the Anti-Cytomegalovirus (Anti-CMV) Immunoglobulin Type M (IgM) Status | Anti-CMV IgM status was assessed by Enzyme-Linked Immunosorbent Assay [ELISA], with respect to positive and negative subjects. Grey Zone = optical density zone within 20% of cut-off value. When an optical density is within this grey zone, sample testing is repeated to confirm the result. | At Month 4 | 
| Anti-CMV Immunoglobulin Type M (IgM) Status, Descriptive Statistics | Anti-CMV IgM status was assessed by Enzyme-Linked Immunosorbent Assay [ELISA], with respect to positive and negative subjects. | At Month 6 | 
| Descriptive Statistics for the Anti-CMV IgM Status | Anti-CMV IgM status was assessed by Enzyme-Linked Immunosorbent Assay [ELISA], with respect to positive and negative subjects. Grey Zone = optical density zone within 20% of cut-off value. When an optical density is within this grey zone, sample testing is repeated to confirm the result. | At pregnancy conclusion (Day 0 to 5, Day 0 = day of delivery, stillbirth or termination) | 
| Anti-glycoprotein B (gB) Immunoglobulin Type G (IgG) Antibody Concentrations | Anti-gB IgG concentrations were assessed by ELISA, presented as geometric mean concentrations (GMCs) and expressed in ELISA units per milliliter (EU/mL). The cut-off value was greater than or equal to (≥) 54 EU/mL. | At Month 0 | 
| Anti-gB IgG Antibody Concentrations | Anti-gB IgG concentrations were assessed by ELISA, presented as geometric mean concentrations (GMCs) and expressed in ELISA units per milliliter (EU/mL). The cut-off value was greater than or equal to (≥) 54 EU/mL. | At pregnancy conclusion (Day 0 to 5, Day 0 = day of delivery, stillbirth or termination) | 
| Descriptive Statistics of the Anti-glycoprotein B (gB) Immunoglobulin Type G (IgG) Avidity Index | The avidity index was calculated as the mean absorbance of reactions in which the immune complexes were exposed to urea divided by the mean absorbance of reactions in which the immune complexes were not exposed to urea, expressed as a percentage. | At Month 0 | 
| Descriptive Statistics of the Anti-gB IgG Avidity Index | The avidity index was calculated as the mean absorbance of reactions in which the immune complexes were exposed to urea divided by the mean absorbance of reactions in which the immune complexes were not exposed to urea, expressed as a percentage. | At pregnancy conclusion (Day 0 to 5, Day 0 = day of delivery, stillbirth or termination) | 
| CMV-specific Cluster of Differentiation 4 (CD4) T-cell Frequencies | Descriptive statistics of the frequency of CMV-specific CD4 T cells expressing at least two markers among: cluster of differentiation 40 ligand (CD40L), interleukin-2 (IL-2), interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), as assessed by Intracellular Cytokine Staining [ICS], by stimulating agent (among Human Cytomegalovirus [HCMV] immediate-early gene [IE1] antigen, HCMV glicoprotein B [gB] antigen, HCMV lysate antigen and HCMV pp65 antigen). | At Month 0 | 
| CMV-specific CD4 T-cell Frequencies | Descriptive statistics of the frequency of CMV-specific CD4 T cells expressing at least two markers among CD40L, IL-2, IFNg, TNFa, as assessed by Intracellular Cytokine Staining [ICS], by stimulating agent (among immediate-early gene [IE1] antigen, glicoprotein B [gB] antigen, CMV lysate antigen and CMV pp65 antigen). | At pregnancy conclusion (Day 0 to 5, Day 0 = day of delivery, stillbirth or termination) | 
| CMV-specific Cluster of Differentiation 8 (CD8) T-cell Frequencies | Descriptive statistics of the frequency of CMV-specific CD8 T cells expressing at least two markers among CD40L, IL-2, IFNg, TNFa, as assessed by Intracellular Cytokine Staining [ICS], by stimulating agent (among immediate-early gene [IE1] antigen, glicoprotein B [gB] antigen, CMV lysate antigen and CMV pp65 antigen). | At Month 0 | 
| CMV-specific CD8 T-cell Frequencies | Descriptive statistics of the frequency of CMV-specific CD8 T cells expressing at least two markers among CD40L, IL-2, IFNg, TNFa, as assessed by Intracellular Cytokine Staining [ICS], by stimulating agent (among immediate-early gene [IE1] antigen, glicoprotein B [gB] antigen, CMV lysate antigen and CMV pp65 antigen). | At pregnancy conclusion (Day 0 to 5, Day 0 = day of delivery, stillbirth or termination) | 
| CMV-specific Proliferating Cluster of Differentiation (CD4) T Cells Frequencies | Labelled cells were quantified by flow cytometry, by stimulating agent (among immediate-early gene [IE1] antigen, glicoprotein B [gB] antigen, CMV lysate antigen and CMV pp65 antigen). | At Month 0 | 
| CMV-specific Proliferating CD4 T Cells Frequencies | Labelled cells were quantified by flow cytometry, by stimulating agent (among immediate-early gene [IE1] antigen, glicoprotein B [gB] antigen, CMV lysate antigen and CMV pp65 antigen). Note: Results were retrieved by subtracting the background without imputing the negative and zero values, this generated negative values. | At pregnancy conclusion (Day 0 to 5, Day 0 = day of delivery, stillbirth or termination) | 
| Concentrations of Anti-CMV Tegument Protein Immunoglobulin G (IgG) Antibodies | Antibody concentrations were expressed as Geometric Mean Concentrations (GMCs), measured in units per milliliter (U/mL). Concentrations of antibodies were assessed by the Enzyme-Linked Immunosorbent Assay (ELISA) for a cut-off greater than or equal to (≥) 0.668 U/mL. | At Day 0 = study entry | 
| Anti-CMV Tegument Protein Immunoglobulin G (IgG) Antibody Concentrations | Antibody concentrations were expressed as Geometric Mean Concentrations (GMCs), measured in units per milliliter (U/mL). Concentrations of antibodies were assessed by the Enzyme-Linked Immunosorbent Assay (ELISA) for a cut-off greater than or equal to (≥) 0.668 U/mL. | At Month 2 | 
| Concentrations of Anti-CMV Tegument Protein IgG Antibodies | Antibody concentrations were expressed as Geometric Mean Concentrations (GMCs), measured in units per milliliter (U/mL). Concentrations of antibodies were assessed by the Enzyme-Linked Immunosorbent Assay (ELISA) for a cut-off greater than or equal to (≥) 0.668 U/mL. | At Month 4 | 
| Anti-CMV Tegument Protein IgG Antibody Concentrations | Antibody concentrations were expressed as Geometric Mean Concentrations (GMCs), measured in units per milliliter (U/mL). Concentrations of antibodies were assessed by the Enzyme-Linked Immunosorbent Assay (ELISA) for a cut-off greater than or equal to (≥) 0.668 U/mL. | At Month 6 | 
| Concentrations of Anti-CMV IgG Antibodies | Antibody concentrations were expressed as Geometric Mean Concentrations (GMCs), measured in units per milliliter (U/mL). Concentrations of antibodies were assessed by the Enzyme-Linked Immunosorbent Assay (ELISA) for a cut-off greater than or equal to (≥) 0.668 U/mL. | At pregnancy conclusion (Day 0 to 5, Day 0 = day of delivery, stillbirth or termination) | 
| Descriptive Statistics of the Anti-CMV Tegument Protein Globulin Type B (gB) Immunoglobulin G (IgG) Avidity, by Congenital Infection Status | Avidity of anti-CMV tegument protein gB IgG was assessed by ELISA. The avidity index was calculated as the mean absorbance of reactions in which the immune complexes were exposed to urea divided by the mean absorbance of reactions in which the immune complexes were not exposed to urea, expressed as a percentage. | At Day 0 = study entry | 
| Descriptive Statistics of the Anti-CMV Tegument Protein gB Immunoglobulin G (IgG) Avidity, by Congenital Infection Status | Avidity of anti-CMV tegument protein gB IgG was assessed by ELISA. The avidity index was calculated as the mean absorbance of reactions in which the immune complexes were exposed to urea divided by the mean absorbance of reactions in which the immune complexes were not exposed to urea, expressed as a percentage. | At Month 2 | 
| Descriptive Statistics of the Anti-CMV Tegument Protein gB IgG Avidity, by Congenital Infection Status | Avidity of anti-CMV tegument protein gB IgG was assessed by ELISA. The avidity index was calculated as the mean absorbance of reactions in which the immune complexes were exposed to urea divided by the mean absorbance of reactions in which the immune complexes were not exposed to urea, expressed as a percentage. | At Month 4 | 
| Descriptive Statistics of the Anti-CMV Tegument Protein Globulin Type B (gB) IgG Avidity, by Congenital Infection Status | Avidity of anti-CMV tegument protein gB IgG was assessed by ELISA. The avidity index was calculated as the mean absorbance of reactions in which the immune complexes were exposed to urea divided by the mean absorbance of reactions in which the immune complexes were not exposed to urea, expressed as a percentage. | At Month 6 | 
| Anti-CMV Tegument Protein Globulin Type B (gB) Immunoglobulin G (IgG) Avidity Descriptive Statistics, by Congenital Infection Status | Avidity of anti-CMV tegument protein gB IgG was assessed by ELISA. The avidity index was calculated as the mean absorbance of reactions in which the immune complexes were exposed to urea divided by the mean absorbance of reactions in which the immune complexes were not exposed to urea, expressed as a percentage. | At pregnancy conclusion (Day 0 to 5, Day 0 = day of delivery, stillbirth or termination) | 
| Anti-CMV Antibody Titers, by Neutralisation Assay (Fibroblast) | Antibody titers were expressed as Geometric Mean Titers (GMTs), for the seropositivity cut-off of ≥ 10 ED50. | At Month 0 | 
| Anti-CMV Antibody Titers, by Neutralisation Assay (Fibroblast) | Antibody titers were expressed as Geometric Mean Titers (GMTs), for the seropositivity cut-off of ≥ 10 ED50. | At Month 2 | 
| Anti-CMV Antibody Titers, by Neutralisation Assay (Fibroblast) | Antibody titers were expressed as Geometric Mean Titers (GMTs), for the seropositivity cut-off of ≥ 10 ED50. | At Month 4 | 
| Anti-CMV Antibody Titers, by Neutralisation Assay (Fibroblast) | Antibody titers were expressed as Geometric Mean Titers (GMTs), for the seropositivity cut-off of ≥ 10 ED50. | At Month 6 | 
| Anti-CMV Antibody Titers, by Neutralisation Assay (Fibroblast) | Antibody titers were expressed as Geometric Mean Titers (GMTs), for the seropositivity cut-off of ≥ 10 ED50. | At pregnancy conclusion (Day 0 to 5, Day 0 = day of delivery, stillbirth or termination) | 
| Anti-CMV Antibody Titers, by Neutralisation Assay (Epithelial Cells) | Antibody titers were expressed as Geometric Mean Titers (GMTs), for the seropositivity cut-off of ≥ 15 ED50. | At Month 0 | 
| Anti-CMV Antibody Titers, by Neutralisation Assay (Epithelial Cells) | Antibody titers were expressed as Geometric Mean Titers (GMTs), for the seropositivity cut-off of ≥ 15 ED50. | At Month 2 | 
| Anti-CMV Antibody Titers, by Neutralisation Assay (Epithelial Cells) | Antibody titers were expressed as Geometric Mean Titers (GMTs), for the seropositivity cut-off of ≥ 15 ED50. | At Month 4 | 
| Anti-CMV Antibody Titers, by Neutralisation Assay (Epithelial Cells) | Antibody titers were expressed as Geometric Mean Titers (GMTs), for the seropositivity cut-off of ≥ 15 ED50. | At Month 6 | 
| Anti-CMV Antibody Titers, by Neutralisation Assay (Epithelial Cells) | Antibody titers were expressed as Geometric Mean Titers (GMTs), for the seropositivity cut-off of ≥ 10 ED50. | At pregnancy conclusion (Day 0 to 5, Day 0 = day of delivery, stillbirth or termination) | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Pregnancy, Congenital, Cytomegalovirus infection