0ba130375558e62bea0b246d4567ed08 pmc_10675013.pdf 6988c48424c36c46623b70cee2b24cf027982223 pmc_10675013.pdf c1ef7cd451b396c7e4aa3424d314ec87d76fd5af5c0ae017add5d8ebde8a7262 pmc_10675013.pdf Title: Vaccination with an HIV T-Cell Immunogen (HTI) Using DNA Primes Followed by a ChAdOx1-MVA Boost Is Immunogenic in Gut Microbiota-Depleted Mice despite Low IL-22 Serum Levels Subject: Despite the important role of gut microbiota in the maturation of the immune system, little is known about its impact on the development of T-cell responses to vaccination. Here, we immunized C57BL/6 mice with a prime-boost regimen using DNA plasmid, the Chimpanzee Adenovirus, and the modified Vaccinia Ankara virus expressing a candidate HIV T-cell immunogen and compared the T-cell responses between individuals with an intact or antibiotic-depleted microbiota. Overall, the depletion of the gut microbiota did not result in significant differences in the magnitude or breadth of the immunogen-specific IFN T-cell response after vaccination. However, we observed marked changes in the serum levels of four cytokines after vaccinating microbiota-depleted animals, particularly a significant reduction in IL-22 levels. Interestingly, the level of IL-22 in serum correlated with the abundance of Roseburia in the large intestine of mice in the mock and vaccinated groups with intact microbiota. This short-chain fatty acid (SCFA)-producing bacterium was significantly reduced in the vaccinated, microbiota-depleted group. Therefore, our results indicate that, although microbiota depletion reduces serum levels of IL-22, the powerful vaccine regime used could have overcome the impact of microbiota depletion on IFN-producing T-cell responses. Keywords: microbiota; HIV; vaccine; T-cell; IFN; IL-22; Roseburia Author: Aleix Elizalde-Torrent, Alessandra Borgognone, Maria Casadellà, Luis Romero-Martin, Tuixent Escribà, Mariona Parera, Yaiza Rosales-Salgado, Jorge Díaz-Pedroza, Francesc Català-Moll, Marc Noguera-Julian, Christian Brander, Roger Paredes and Alex Olvera Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Tue Oct 31 07:31:40 2023 CET ModDate: Tue Oct 31 07:34:23 2023 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 17 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1730130 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- PYPZKK+VnURWPalladioL-Bold Type 1 Custom yes yes no 10 0 AEPYJE+VnURWPalladioL Type 1 Custom yes yes no 15 0 KNPFEX+EURM10 Type 1 Builtin yes yes no 19 0 URCSMW+URWPalladioL-Bold Type 1 Custom yes yes no 23 0 IHCQPK+URWPalladioL-Roma Type 1 Custom yes yes no 28 0 QCZMWR+URWPalladioL-Ital Type 1 Custom yes yes no 32 0 IFEDSM+CMSY10 Type 1 Builtin yes yes no 72 0 JJCBDJ+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 86 0 JJCBDK+PalatinoLinotype CID TrueType Identity-H yes yes yes 89 0 JJCBDL+PalatinoLinotype TrueType WinAnsi yes yes no 95 0 JJCBDN+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 98 0 JJCBPK+PalatinoLinotype TrueType MacRoman yes yes no 101 0 GEWEQC+PazoMath-Italic Type 1 Builtin yes yes no 111 0 JJCEIN+Arial CID TrueType Identity-H yes yes yes 128 0 JJCBDJ+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 134 0 JJCBDK+PalatinoLinotype CID TrueType Identity-H yes yes yes 137 0 JJCBDL+PalatinoLinotype TrueType WinAnsi yes yes no 143 0 JJCBDN+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 146 0 JJCBPK+PalatinoLinotype TrueType MacRoman yes yes no 149 0 JJCEEN+Arial,Bold CID TrueType Identity-H yes yes yes 152 0 JJCBDJ+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 220 0 JJCBDK+PalatinoLinotype CID TrueType Identity-H yes yes yes 223 0 JJCBDL+PalatinoLinotype TrueType WinAnsi yes yes no 229 0 JJCBDM+PalatinoLinotype,Italic CID TrueType Identity-H yes yes yes 232 0 JJCBDN+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 238 0 JJCBPK+PalatinoLinotype TrueType MacRoman yes yes no 241 0 JJCBDJ+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 256 0 JJCBDK+PalatinoLinotype CID TrueType Identity-H yes yes yes 259 0 JJCBDL+PalatinoLinotype TrueType WinAnsi yes yes no 265 0 JJCBDM+PalatinoLinotype,Italic CID TrueType Identity-H yes yes yes 268 0 JJCBDN+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 274 0 JJCBPK+PalatinoLinotype TrueType MacRoman yes yes no 277 0 BBETNX+PazoMath Type 1 Builtin yes yes no 303 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-06-21 02:48:26 CEST RepresentationInformation: pmc_10675013.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-06-20 02:37:25 CEST Size: 1730130 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 578 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Experimental Design Destination: subsection.2.1 Item: Title: Sample Collection and Processing Destination: subsection.2.2 Item: Title: DNA Extraction and 16S rRNA Sequencing Destination: subsection.2.3 Item: Title: 16S rRNA Sequence Analysis Destination: subsection.2.4 Item: Title: Overlapping Peptides Covering the HTI Sequence Destination: subsection.2.5 Item: Title: Mouse IFN ELISPOT Assay Destination: subsection.2.6 Item: Title: Luminex Assay Destination: subsection.2.7 Item: Title: Statistical Analysis of Vaccine Response Data Destination: subsection.2.8 Item: Title: Results Destination: section.3 Children: Item: Title: Vaccination with a Highly Immunogenic Prime-Boost Strategy Triggers Strong HTI-Specific IFN Producing T-Cell Responses despite Depleted Microbiota Destination: subsection.3.1 Item: Title: Gut Microbiota and DDDCM.HTI Vaccination Affect Serum Cytokine Levels Destination: subsection.3.2 Item: Title: IL-22 Serum Levels Correlate with the Abundance of Short Chain Fatty Acid (SCFA) Producing Bacteria in Large Intestine Destination: subsection.3.3 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.A. Info: Title: Vaccination with an HIV T-Cell Immunogen (HTI) Using DNA Primes Followed by a ChAdOx1-MVA Boost Is Immunogenic in Gut Microbiota-Depleted Mice despite Low IL-22 Serum Levels Author: Aleix Elizalde-Torrent, Alessandra Borgognone, Maria Casadellà, Luis Romero-Martin, Tuixent Escribà, Mariona Parera, Yaiza Rosales-Salgado, Jorge Díaz-Pedroza, Francesc Català-Moll, Marc Noguera-Julian, Christian Brander, Roger Paredes and Alex Olvera Subject: Despite the important role of gut microbiota in the maturation of the immune system, little is known about its impact on the development of T-cell responses to vaccination. Here, we immunized C57BL/6 mice with a prime-boost regimen using DNA plasmid, the Chimpanzee Adenovirus, and the modified Vaccinia Ankara virus expressing a candidate HIV T-cell immunogen and compared the T-cell responses between individuals with an intact or antibiotic-depleted microbiota. Overall, the depletion of the gut microbiota did not result in significant differences in the magnitude or breadth of the immunogen-specific IFN T-cell response after vaccination. However, we observed marked changes in the serum levels of four cytokines after vaccinating microbiota-depleted animals, particularly a significant reduction in IL-22 levels. Interestingly, the level of IL-22 in serum correlated with the abundance of Roseburia in the large intestine of mice in the mock and vaccinated groups with intact microbiota. This short-chain fatty acid (SCFA)-producing bacterium was significantly reduced in the vaccinated, microbiota-depleted group. Therefore, our results indicate that, although microbiota depletion reduces serum levels of IL-22, the powerful vaccine regime used could have overcome the impact of microbiota depletion on IFN-producing T-cell responses. 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