Research Project 1: Detection of polyreactive antibodies in saliva and breastmilk
- Thematic field of study: Detection of polyreactive antibodies in saliva and breastmilk
- Supervisor(s):Prof Clive Gray, Dr Helen Payne, Dr Novel Chegou, Dr Lily Verhagen
- Contact details: cgray@sun.ac.za Professor Clive Gray
Summary of hosting Lab
| The Gray lab at Stellenbosh University works on Immune Regulation in Maternal and Child Health in the context of Reproductive Immunology. Prof Gray investigates the characteristics and functional capacity of T cells and other cell types in HIV exposed, uninfected newborn infants. This has great relevance for understanding abnormalities in immune regulatory networks and will provide clues to understanding susceptibility to HIV infection and in devising new treatments that can reduce the morbidities found in this vulnerable infant population. His team studies the human placenta and the impact of maternal HIV infection on cells at the feto-maternal interface. He has recently established the Reproductive Immunology Research Consortium in Africa at Stellenbosch University (SU). |
Project Description
| Our earlier research has highlighted the significance of low levels of immunoglobulin A (IgA), an antibody that is abundantly present in saliva, as a risk factor for respiratory tract infections in children. Both IgA and IgG antibodies are detectable in mucosal samples. Recent findings show a pool of respiratory mucosal broadly neutralizing IgA and IgG antibodies that naturally bind to multiple targets and act as the initial defense against new pathogens. Polyreactive mucosal antibodies play an important role in the susceptibility to and transmission of respiratory viruses. However, it remains unknown whether these antibodies are present in early life and if mothers transfer them to their infants through breastmilk. In this project, we will therefore develop an assay capable of detecting polyreactive antibodies in both infant saliva and maternal breastmilk and examine the relationship with maternal and infant characteristics, including HIV exposure. This assay will utilize a Luminex Multiplex Immuno Assay, a robust tool for quantifying antibodies in low-volume samples. The assay employs fluorescently labeled microspheres coated with five structurally and biologically distinct antigens, binding the same subset of broadly reactive antibodies.. |
Publications
| Koenen, M. H., van Montfrans, J. M., Sanders, E. A. M., Bogaert, D. & Verhagen, L. M. Immunoglobulin A deficiency in children, an undervalued clinical issue. Clin. Immunol. 209, 108293 (2019). Koenen, M. et al. A novel method to standardise serum IgA measurements shows an increased prevalence of IgA deficiency in young children with recurrent respiratory tract infections. Clin. Transl. Immunol. 10, e1344 (2021).. Koenen, M. H. et al. Articles Mycoplasma pneumoniae carriage in children with recurrent respiratory tract infections is associated with a less diverse and altered microbiota. eBioMedicine 98, 104868 (2023).. Gallo, O., Locatello, L. G., Mazzoni, A., Novelli, L. & Annunziato, F. The central role of the nasal microenvironment in the transmission, modulation, and clinical progression of SARS-CoV-2 infection. Mucosal Immunol. 14, 305–316 (2021). Verhagen, L. Respiratory tract infections: importance of the mucosal immune system. Eur. Soc. Paediatr. Infect. Dis. Conf. 2022. Plenary Symp. Present. Available from https//www.youtube.com/watch?v=eJXrNpQB1nU.. |
Major Lab techniques
| Sample processing, Luminex Multiplex Immuno Assay setup, Fluorescent microsphere handling, Collaborative research, Documentation and reporting, Data analysis |
Research Project 2: CMV infection and protective immunity at the maternal-fetal interface in human
- Thematic field of study: CMV infection and protective immunity at the maternal-fetal interface in human
- Supervisor(s): Dr. Doty Ojwach, Dr. Helen Payne and Professor Clive Gray
- Contact details: cgray@sun.ac.za Professor Clive Gray
Summary of hosting Lab
| The Gray lab at Stellenbosh University works on Immune Regulation in Maternal and Child Health in the context of Reproductive Immunology. Prof Gray investigates the characteristics and functional capacity of T cells and other cell types in HIV exposed, uninfected newborn infants. This has great relevance for understanding abnormalities in immune regulatory networks and will provide clues to understanding susceptibility to HIV infection and in devising new treatments that can reduce the morbidities found in this vulnerable infant population. His team studies the human placenta and the impact of maternal HIV infection on cells at the feto-maternal interface. He has recently established the Reproductive Immunology Research Consortium in Africa at Stellenbosch University (SU). |
Project Description
| Cytomegalovirus (CMV) reactivation frequently occurs during gestation especially in HIV-1 infected women including those on suppressive ART. In some African clinics, CMV infection is universal with >90% of pregnant women with serostatus compatible with non-primary CMV infection. CMV has been reported to enhance placental cell susceptibility to replication of HIV-1, and may facilitate in utero HIV transmission. We hypothesize that maternal HIV and CMV co-infection disrupts immune homeostatic balance at the foetal-maternal interface (FMI) leading to dysregulation in infant immunity. In vitro, CMV traditionally infect a limited range of cells such as skin fibroblasts, whereas in vivo, these viruses exhibit a broader cell tropism and CMV dissemination is believed to occur primarily through cell-associated virus, rather than free virus particles. We wish in this project to examine possible protective immunity at the maternal-fetal interface (MFI) by examining for T cell immunity in the decidua membranes of placentas at delivery. We predict that high levels of T cell responsiveness to CMV peptides/epitopes at the MFI will negatively associate with the presence of CMV in the placenta, and vice versa. This project aims has 2 objectives: 1. To identify CMV-specific T cells in the decidua 2. To measure CMV infection in the placenta by identifying anti-pp65 and anti-IE proteins in specific placental cells, employing techniques such as immunohistochemistry, multiplex immunofluorescence and qPCR. By employing established protocols, this project will lay the foundation for gaining mechanistic insights into the relationship between immune events at the FMI, ultimately contributing to our understanding of the relationship between maternal-foetal interactions and compromised neonatal immunity. |
Publications
| Alfi O, Cohen M, Bar-On S, Hashimshony T, Levitt L, Raz Y, Blecher Y, Chaudhry MZ, Cicin-Sain L, Ben-El R, Oiknine-Djian E, Lahav T, Vorontsov O, Cohen A, Zakay-RonesZ, Daniel L, Berger M, Mandel-Gutfreund Y, Panet A, Wolf DG. Decidual-tissue-resident memory T cells protect against nonprimary human cytomegalovirus infection at the maternal-fetal interface. Cell Rep. 2024 Jan 23;43(2):113698. doi: 10.1016/j.celrep.2024.113698. Epub ahead of print. PMID: 38265934. Kim Y, Kim YM, Kim DR, Kim HG, Sung JH, Choi SJ, Oh SY, Kim YJ, Chang YS, Kim D, Kim JS, Moon IJ, Roh CR. The Multifaceted Clinical Characteristics of Congenital Cytomegalovirus Infection: From Pregnancy to Long-Term Outcomes. J Korean Med Sci. 2023 Aug 14;38(32):e249. doi: 10.3346/jkms.2023.38.e249. PMID: 37582499; PMCID: PMC10427218. Degrelle SA, Buchrieser J, Dupressoir A, Porrot F, Loeuillet L, Schwartz O, Fournier T. IFITM1 inhibits trophoblast invasion and is induced in placentas associated with IFN- mediated pregnancy diseases. iScience. 2023 Jun 15;26(7):107147. doi: 10.1016/j.isci.2023.107147. PMID: 37434700; PMCID: PMC10331461. Girsch JH, Mejia Plazas MC, Olivier A, Farah M, Littlefield D, Behl S, Punia S, Sakemura R, Hemsath JR, Norgan A, Enninga EAL, Johnson EL, Chakraborty R. Host- Viral Interactions at the Maternal-Fetal Interface. What We Know and What We Need to Know. Frontiers (Boulder). 2022 Mar;2:833106. doi: 10.3389/fviro.2022.833106. Epub 2022 Mar 24. PMID: 36742289; PMCID: PMC9894500. Forner G, Saldan A, Mengoli C, Pizzi S, Fedrigo M, Gussetti N, Visentin S, Angelini A, Cosmi E, Barzon L, Abate DA. Four-Year Follow-Up of the Maternal Immunological, Virological and Clinical Settings of a 36-Year-Old Woman Experiencing Primary Cytomegalovirus Infection Leading to Intrauterine Infection. Viruses. 2022 Dec 30;15(1):112. doi: 10.3390/v15010112. PMID: 36680152; PMCID: PMC9865737. |
Major Lab techniques
| Cell culture, Histology (cell and tissue staining), Immune cell quantification in tissue, dissection and sectioning of placenta, ELISA and qPCR. |
Research Project 3:The role of Glucagon-like peptide-1 (GLP-1) in placental immune responses
- Thematic field of study: The role of Glucagon-like peptide-1 (GLP-1) in placental immune responses
- Supervisor(s):Dr Tariq Webber, Prof Clive Gray
- Contact details: twebber@sun.ac.za
Summary of hosting Lab
The Gray lab at Stellenbosh University works on Immune Regulation in Maternal and Child Health in the context of Reproductive Immunology. Prof Gray investigates the characteristics and functional capacity of T cells and other cell types in HIV exposed, uninfected newborn infants. This has great relevance for understanding abnormalities in immune regulatory networks and will provide clues to understanding susceptibility to HIV infection and in devising new treatments that can reduce the morbidities found in this vulnerable infant population. His team studies the human placenta and the impact of maternal HIV infection on cells at the feto-maternal interface. This project will be based at the Reproductive Immunology Research Consortium in Africa at Stellenbosch University (SU).
Project Description
HIV and ARTs are known to cause poor birth outcomes, which could be placenta driven. Inflammation in the placenta is a known effect of ARTs, but the mechanism is unclear. Metabolic dysfunction is also a known effect of ARTs, either via disruption of hormones or the immune response. We propose that metabolic hormones play a role in translating metabolic disruptions from ARTs through to cells in the placenta, resulting in inflammation. One of these hormones, Glucagon-like peptide-1 (GLP-1), is produced in the small intestine and plays crucial roles in metabolic regulation systemically and in immune cells, and has anti-inflammatory effects. GLP-1 concentrations decrease in maternal serum as pregnancy progresses, but are decreased during HIV infection and ART. While contra-indication for GLP-1 agonist use during pregnancy is based on a lack of sufficient human experimental evidence for its safety, its use in mouse studies displayed reduced foetal weight or growth and delayed bone development. The role of GLP-1 in placental health and function is unclear and its effect on the inflammatory responses in the placenta remain unclear. We hypothesise that GLP-1 will stimulate anti-inflammatory responses in placental lymphocytes.
Aim:
To demonstrate immunomodulatory properties of GLP-1 in placental cells.
Objectives:
- Isolate lymphocytes from the placenta and peripheral blood.
- Stimulate the cells with GLP-1 and the GLP-1R blocker Exendin 9-39 as control.
- Assess the response to GLP-1 stimulation using flow cytometry to measure:
- intracellular cytokine production (IL-10, IL-6, and TNFα)
- proliferation (Ki-67)
- apoptosis (Annexin V, PI).
Publications
Jurkowska K, Szymańska B, Knysz B, Piwowar A. Effect of Combined Antiretroviral Therapy on the Levels of Selected Parameters Reflecting Metabolic and Inflammatory Disturbances in HIV-Infected Patients. J Clin Med. 2022 Mar 19;11(6):1713. doi: 10.3390/jcm11061713. PMID: 35330038; PMCID: PMC8954290.
Inès Auclair Mangliar, Anne-Sophie Plante, Myriam Chabot, Claudia Savard, Simone Lemieux, Andréanne Michaud, S. John Weisnagel, Félix Camirand Lemyre, Alain Veilleux, and Anne-Sophie Morisset. 2024. GLP-1 response during pregnancy: variations between trimesters andassociations with appetite sensations and usual energy intake. Applied Physiology, Nutrition, and Metabolism. 49(4): 428-436. https://doi.org/10.1139/apnm-2023-0301
Muller DRP, Stenvers DJ, Malekzadeh A, Holleman F, Painter RC, Siegelaar SE. Effects of GLP-1 agonists and SGLT2 inhibitors during pregnancy and lactation on offspring outcomes: a systematic review of the evidence. Front Endocrinol (Lausanne). 2023 Oct 10;14:1215356. doi: 10.3389/fendo.2023.1215356. PMID: 37881498; PMCID: PMC10597691.
Al-Dwairi A, Alqudah TE, Al-Shboul O, Alqudah M, Mustafa AG, Alfaqih MA. Glucagon-like peptide-1 exerts anti-inflammatory effects on mouse colon smooth muscle cells through the cyclic adenosine monophosphate/nuclear factor-κB pathway in vitro. J Inflamm Res. 2018 Mar 20;11:95-109. doi: 10.2147/JIR.S152835. Erratum in: J Inflamm Res. 2019 Sep 26;12:267. doi: 10.2147/JIR.S230818. PMID: 29593427; PMCID: PMC5865574.
Marx N, Burgmaier M, Heinz P, Ostertag M, Hausauer A, Bach H, Durst R, Hombach V, Walcher D. Glucagon-like peptide-1(1-37) inhibits chemokine-induced migration of human CD4-positive lymphocytes. Cell Mol Life Sci. 2010 Oct;67(20):3549-55. doi: 10.1007/s00018-010-0396-5. Epub 2010 May 21. PMID: 20495843; PMCID: PMC11115661.
Major Lab techniques
Sample processing, Placenta dissection, Cell Culture, Lymphocyte Isolation, Stimulation assays, intracellular flow cytometry, proliferation assays, apoptosis analysis, Flow cytometry data analysis, and R programming.
Research Project 4: Investigating F13A1 Expression and Its Role in Macrophage Polarization and Function
- Thematic field of study: Investigating F13A1 Expression and Its Role in Macrophage Polarization and Function
- Supervisor(s): Dr. Doty Ojwach and Professor Clive Gray
- Contact details: cgray@sun.ac.za , Professor Clive Gray
Summary of Hosting Lab
The Gray lab at Stellenbosh University works on Immune Regulation in Maternal and Child Health in the context of Reproductive Immunology. Prof Gray investigates the characteristics and functional capacity of T cells and other cell types in HIV exposed, uninfected newborn infants. This has great relevance for understanding abnormalities in immune regulatory networks and will provide clues to understanding susceptibility to HIV infection and in devising new treatments that can reduce the morbidities found in this vulnerable infant population. His team studies the human placenta and the impact of maternal HIV infection on cells at the feto-maternal interface. This project will be based in the Reproductive Immunology Research Consortium in Africa at Stellenbosch University (SU).
Project Description
Coagulation factor XIII A1 (F13A1) is a key player in immune regulation, particularly in macrophage function. Despite its importance, the differential expression of F13A1 in M1 and M2 macrophages and its potential implications for placental health, particularly in the context of HIV, antiretroviral therapy (ART), and pre-eclampsia, remain underexplored. This project aims to investigate the role of F13A1 in macrophage polarization and its potential link to placental pathologies.
Objectives:
- Characterize F13A1 expression in M1 and M2 macrophages.
- Explore how F13A1 modulates macrophage polarization and function in vitro.
- Investigate the association between F13A1 expression and placental pathologies, specifically pre-eclampsia, in HIV+ and ART-exposed pregnancies.
Methodology:
- Cell Culture: Monocytes will be polarized into M1 and M2 macrophages.
- Western Blot: F13A1 protein expression will be quantified in each macrophage phenotype.
- RT-PCR: Quantify F13A1 mRNA levels in plasma and Hofbauer cells.
- Sequencing: Sequence the F13A1 gene from plasma to assess genetic variation.
- Functional Assays: Macrophage phagocytic activity, cytokine production, and iron metabolism will be assessed in cells with altered F13A1 expression.
- Placental Tissue Analysis: F13A1 expression will be examined in placental macrophages from HIV+ and HIV- pre-eclamptic samples.
Expected Outcomes:
This study will elucidate the role of F13A1 in macrophage polarization and function, and its potential contribution to placental health, particularly in the context of HIV and pre-eclampsia. The findings may offer novel insights for therapeutic strategies targeting macrophage regulation in these conditions.
Publications
| TTukei VJ, Hoffman HJ, Greenberg L, Thabelo R, Nchephe M, Mots’oane T, Masitha M, Chabela M, Mokone M, Mofenson L, Guay L, Tiam A. 2021. Adverse Pregnancy Outcomes Among HIV-positive Women in the Era of Universal Antiretroviral Therapy Remain Elevated Compared With HIV-negative Women. Pediatr Infect Dis J 40:821-826.Sun F, Wang S, Du M. 2021. Functional regulation of decidual macrophages during pregnancy. Journal of Reproductive Immunology 143:103264.
Alshehri FSM, Whyte CS, Mutch NJ. 2021. Factor XIII-A: An Indispensable “Factor” in Haemostasis and Wound Healing. Int J Mol Sci 22. Cordell PA, Kile BT, Standeven KF, Josefsson EC, Pease RJ, Grant PJ. 2010. Association of coagulation factor XIII-A with Golgi proteins within monocyte-macrophages: implications for subcellular trafficking and secretion. Blood, The Journal of the American Society of Hematology 115:2674-2681. Töröcsik D, Széles L, Paragh Jr G, Rákosy Z, Bárdos H, Nagy L, Balázs M, Inbal A, Ádány R. 2010. Factor XIII-A is involved in the regulation of gene expression in alternatively activated human macrophages. Thrombosis and haemostasis 104:709-717. Martinez FO, Gordon S, Locati M, Mantovani A. 2006. Transcriptional Profiling of the Human Monocyte-to-Macrophage Differentiation and Polarization: New Molecules and Patterns of Gene Expression1. The Journal of Immunology 177:7303-7311. |
Major Lab techniques
Cell culture, Histology (cell and tissue staining), western blot, dissection and sectioning of placenta, ELISA, qPCR and data analysis.
Links to other University Program
https://www.sun.ac.za/english/faculty/healthsciences/Molecular_Biology_Human_Genetics/Pages/MSc.aspx
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