379 Characterizing the single-cell transcriptomes of fetal natural killer cells isolated from the umbilical cord of fetuses exposed to human cytomegalovirus during gestation
OBJECTIVES/GOALS: Congenital cytomegalovirus (cCMV) remains to be the leading infectious cause of fetal anomalies. The role of fetal natural killer (NK) cells during cCMV remains largely unknown. The objective of this study is to define the transcriptomes of fetal NK cells exposed to human cytomegal...
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| Format: | Article |
| Language: | English |
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Cambridge University Press
2024-04-01
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| Series: | Journal of Clinical and Translational Science |
| Online Access: | https://www.cambridge.org/core/product/identifier/S2059866124003339/type/journal_article |
| _version_ | 1797226913666695168 |
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| author | Mohamed Khalil Scott Terhune Subramaniam Malarkannan |
| author_facet | Mohamed Khalil Scott Terhune Subramaniam Malarkannan |
| author_sort | Mohamed Khalil |
| collection | DOAJ |
| description | OBJECTIVES/GOALS: Congenital cytomegalovirus (cCMV) remains to be the leading infectious cause of fetal anomalies. The role of fetal natural killer (NK) cells during cCMV remains largely unknown. The objective of this study is to define the transcriptomes of fetal NK cells exposed to human cytomegalovirus (HCMV) infection during gestation. METHODS/STUDY POPULATION: Four sets of umbilical cord blood and matching umbilical cord tissues were collected from two HCMV seropositive (HCMV+) and two HCMV seronegative (HCMV-) fetuses that did not experience any complications during gestation. These samples were provided by the Medical College of Wisconsin Tissue Bank and were processed within 24 hours following live birth. CD7+ CD3e-CD14-CD19-CD20- fetal NK cells were isolated, using the BD FACSAria sorter. Following cell sorting, single-cell RNA sequencing (scRNA-seq) was performed, and cDNA libraries were constructed and sequenced via NextSeq 550. Cell Ranger was then used to algin the cDNA reads and the Seurat R package was used to analyze the transcriptional data. Cells were filtered and clustered based on the number of uniquely expressed genes. RESULTS/ANTICIPATED RESULTS: Four sets of umbilical cord blood and matching umbilical cord tissues were collected from two HCMV+ and two HCMV- fetuses. We were able to successfully sort and capture fetal NK cells and perform scRNA-seq on these samples. Following unbiased clustering, we observed and characterized five distinct fetal NK cell subsets in the umbilical cord blood and four fetal NK cell subsets in the corresponding umbilical cord tissue. Our findings revealed that HCMV+ fetal NK cells primarily consisted of mature NK cell subsets, while HCMV- fetal NK cells constituted the majority of the immature subsets. Importantly, we identified a unique subset of NKG2CHi fetal NK cells that were significantly elevated in the HCMV+ fetuses. Finally, we defined a group of transcription factors involved in the formation of antiviral fetal NK. DISCUSSION/SIGNIFICANCE: Here, we demonstrate that HCMV infection can induce the formation of distinct NK cell subsets and drive their unique transcriptional profiles. These findings have the potential to guide the development of an innovative NK cell immunotherapy that could help prevent fetuses from developing symptomatic cCMV. |
| first_indexed | 2024-04-24T14:32:28Z |
| format | Article |
| id | doaj.art-d373da10d79f4a47b60e018a112bed23 |
| institution | Directory Open Access Journal |
| issn | 2059-8661 |
| language | English |
| last_indexed | 2024-04-24T14:32:28Z |
| publishDate | 2024-04-01 |
| publisher | Cambridge University Press |
| record_format | Article |
| series | Journal of Clinical and Translational Science |
| spelling | doaj.art-d373da10d79f4a47b60e018a112bed232024-04-03T02:00:21ZengCambridge University PressJournal of Clinical and Translational Science2059-86612024-04-01811311410.1017/cts.2024.333379 Characterizing the single-cell transcriptomes of fetal natural killer cells isolated from the umbilical cord of fetuses exposed to human cytomegalovirus during gestationMohamed Khalil0Scott Terhune1Subramaniam Malarkannan2Laboratory of Molecular Immunology and Immunotherapy, Blood Research Institute, Versiti, Milwaukee, WI , USA Department of Microbiology and Immunology, Medical College of Wisconsin, Milwaukee, WI , USADepartment of Microbiology and Immunology, Medical College of Wisconsin, Milwaukee, WI , USALaboratory of Molecular Immunology and Immunotherapy, Blood Research Institute, Versiti, Milwaukee, WI , USA Department of Microbiology and Immunology, Medical College of Wisconsin, Milwaukee, WI , USA Department of Pediatrics, Medical College of Wisconsin, Milwaukee, WI , USA Department of Medicine, Medical College of Wisconsin, Milwaukee, WI , USAOBJECTIVES/GOALS: Congenital cytomegalovirus (cCMV) remains to be the leading infectious cause of fetal anomalies. The role of fetal natural killer (NK) cells during cCMV remains largely unknown. The objective of this study is to define the transcriptomes of fetal NK cells exposed to human cytomegalovirus (HCMV) infection during gestation. METHODS/STUDY POPULATION: Four sets of umbilical cord blood and matching umbilical cord tissues were collected from two HCMV seropositive (HCMV+) and two HCMV seronegative (HCMV-) fetuses that did not experience any complications during gestation. These samples were provided by the Medical College of Wisconsin Tissue Bank and were processed within 24 hours following live birth. CD7+ CD3e-CD14-CD19-CD20- fetal NK cells were isolated, using the BD FACSAria sorter. Following cell sorting, single-cell RNA sequencing (scRNA-seq) was performed, and cDNA libraries were constructed and sequenced via NextSeq 550. Cell Ranger was then used to algin the cDNA reads and the Seurat R package was used to analyze the transcriptional data. Cells were filtered and clustered based on the number of uniquely expressed genes. RESULTS/ANTICIPATED RESULTS: Four sets of umbilical cord blood and matching umbilical cord tissues were collected from two HCMV+ and two HCMV- fetuses. We were able to successfully sort and capture fetal NK cells and perform scRNA-seq on these samples. Following unbiased clustering, we observed and characterized five distinct fetal NK cell subsets in the umbilical cord blood and four fetal NK cell subsets in the corresponding umbilical cord tissue. Our findings revealed that HCMV+ fetal NK cells primarily consisted of mature NK cell subsets, while HCMV- fetal NK cells constituted the majority of the immature subsets. Importantly, we identified a unique subset of NKG2CHi fetal NK cells that were significantly elevated in the HCMV+ fetuses. Finally, we defined a group of transcription factors involved in the formation of antiviral fetal NK. DISCUSSION/SIGNIFICANCE: Here, we demonstrate that HCMV infection can induce the formation of distinct NK cell subsets and drive their unique transcriptional profiles. These findings have the potential to guide the development of an innovative NK cell immunotherapy that could help prevent fetuses from developing symptomatic cCMV.https://www.cambridge.org/core/product/identifier/S2059866124003339/type/journal_article |
| spellingShingle | Mohamed Khalil Scott Terhune Subramaniam Malarkannan 379 Characterizing the single-cell transcriptomes of fetal natural killer cells isolated from the umbilical cord of fetuses exposed to human cytomegalovirus during gestation Journal of Clinical and Translational Science |
| title | 379 Characterizing the single-cell transcriptomes of fetal natural killer cells isolated from the umbilical cord of fetuses exposed to human cytomegalovirus during gestation |
| title_full | 379 Characterizing the single-cell transcriptomes of fetal natural killer cells isolated from the umbilical cord of fetuses exposed to human cytomegalovirus during gestation |
| title_fullStr | 379 Characterizing the single-cell transcriptomes of fetal natural killer cells isolated from the umbilical cord of fetuses exposed to human cytomegalovirus during gestation |
| title_full_unstemmed | 379 Characterizing the single-cell transcriptomes of fetal natural killer cells isolated from the umbilical cord of fetuses exposed to human cytomegalovirus during gestation |
| title_short | 379 Characterizing the single-cell transcriptomes of fetal natural killer cells isolated from the umbilical cord of fetuses exposed to human cytomegalovirus during gestation |
| title_sort | 379 characterizing the single cell transcriptomes of fetal natural killer cells isolated from the umbilical cord of fetuses exposed to human cytomegalovirus during gestation |
| url | https://www.cambridge.org/core/product/identifier/S2059866124003339/type/journal_article |
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