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(A) Schematic representation of inoculation protocol. C57Bl/6 mice were injected intravenously or intramuscularly with PBS, peptide, 500 nm SSHEL, or 500 nm SSHEL EncPep and boosted via the same inoculation route 2 weeks later. After 1 week, blood sample was collected, CD3e/CD8a double positive cytotoxic T cells were isolated by flow cytometry and analyzed for their ability to bind to <t>MHC-I/SIINFEKL.</t> (B) Representative flow cytometry plot of isolated CD3e/CD8a positive from mice inoculated intramuscularly with 500 nm SSHEL EncPep that recognize the <t>SIINFEKL</t> <t>tetramer.</t> (C) Percentage of CD3e/CD8a positive cytotoxic T cells that recognize SIINFEKL when presented in MHC-I harvested from mice injected (left) intravenously or (right) intramuscularly with (black) PBS, (pink) free peptide, (blue) SSHEL, or (green) SSHEL EncPep .
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(A) Schematic representation of inoculation protocol. C57Bl/6 mice were injected intravenously or intramuscularly with PBS, peptide, 500 nm SSHEL, or 500 nm SSHEL EncPep and boosted via the same inoculation route 2 weeks later. After 1 week, blood sample was collected, CD3e/CD8a double positive cytotoxic T cells were isolated by flow cytometry and analyzed for their ability to bind to <t>MHC-I/SIINFEKL.</t> (B) Representative flow cytometry plot of isolated CD3e/CD8a positive from mice inoculated intramuscularly with 500 nm SSHEL EncPep that recognize the <t>SIINFEKL</t> <t>tetramer.</t> (C) Percentage of CD3e/CD8a positive cytotoxic T cells that recognize SIINFEKL when presented in MHC-I harvested from mice injected (left) intravenously or (right) intramuscularly with (black) PBS, (pink) free peptide, (blue) SSHEL, or (green) SSHEL EncPep .
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(A) Schematic representation of inoculation protocol. C57Bl/6 mice were injected intravenously or intramuscularly with PBS, peptide, 500 nm SSHEL, or 500 nm SSHEL EncPep and boosted via the same inoculation route 2 weeks later. After 1 week, blood sample was collected, CD3e/CD8a double positive cytotoxic T cells were isolated by flow cytometry and analyzed for their ability to bind to <t>MHC-I/SIINFEKL.</t> (B) Representative flow cytometry plot of isolated CD3e/CD8a positive from mice inoculated intramuscularly with 500 nm SSHEL EncPep that recognize the <t>SIINFEKL</t> <t>tetramer.</t> (C) Percentage of CD3e/CD8a positive cytotoxic T cells that recognize SIINFEKL when presented in MHC-I harvested from mice injected (left) intravenously or (right) intramuscularly with (black) PBS, (pink) free peptide, (blue) SSHEL, or (green) SSHEL EncPep .
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(A) Schematic representation of inoculation protocol. C57Bl/6 mice were injected intravenously or intramuscularly with PBS, peptide, 500 nm SSHEL, or 500 nm SSHEL EncPep and boosted via the same inoculation route 2 weeks later. After 1 week, blood sample was collected, CD3e/CD8a double positive cytotoxic T cells were isolated by flow cytometry and analyzed for their ability to bind to <t>MHC-I/SIINFEKL.</t> (B) Representative flow cytometry plot of isolated CD3e/CD8a positive from mice inoculated intramuscularly with 500 nm SSHEL EncPep that recognize the <t>SIINFEKL</t> <t>tetramer.</t> (C) Percentage of CD3e/CD8a positive cytotoxic T cells that recognize SIINFEKL when presented in MHC-I harvested from mice injected (left) intravenously or (right) intramuscularly with (black) PBS, (pink) free peptide, (blue) SSHEL, or (green) SSHEL EncPep .
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(A) Schematic representation of inoculation protocol. C57Bl/6 mice were injected intravenously or intramuscularly with PBS, peptide, 500 nm SSHEL, or 500 nm SSHEL EncPep and boosted via the same inoculation route 2 weeks later. After 1 week, blood sample was collected, CD3e/CD8a double positive cytotoxic T cells were isolated by flow cytometry and analyzed for their ability to bind to <t>MHC-I/SIINFEKL.</t> (B) Representative flow cytometry plot of isolated CD3e/CD8a positive from mice inoculated intramuscularly with 500 nm SSHEL EncPep that recognize the <t>SIINFEKL</t> <t>tetramer.</t> (C) Percentage of CD3e/CD8a positive cytotoxic T cells that recognize SIINFEKL when presented in MHC-I harvested from mice injected (left) intravenously or (right) intramuscularly with (black) PBS, (pink) free peptide, (blue) SSHEL, or (green) SSHEL EncPep .
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(A) Schematic representation of inoculation protocol. C57Bl/6 mice were injected intravenously or intramuscularly with PBS, peptide, 500 nm SSHEL, or 500 nm SSHEL EncPep and boosted via the same inoculation route 2 weeks later. After 1 week, blood sample was collected, CD3e/CD8a double positive cytotoxic T cells were isolated by flow cytometry and analyzed for their ability to bind to MHC-I/SIINFEKL. (B) Representative flow cytometry plot of isolated CD3e/CD8a positive from mice inoculated intramuscularly with 500 nm SSHEL EncPep that recognize the SIINFEKL tetramer. (C) Percentage of CD3e/CD8a positive cytotoxic T cells that recognize SIINFEKL when presented in MHC-I harvested from mice injected (left) intravenously or (right) intramuscularly with (black) PBS, (pink) free peptide, (blue) SSHEL, or (green) SSHEL EncPep .

Journal: bioRxiv

Article Title: Immunization with peptide encapsulated within synthetic spores activates T cell responses and reduces tumor growth

doi: 10.1101/2025.02.27.640614

Figure Lengend Snippet: (A) Schematic representation of inoculation protocol. C57Bl/6 mice were injected intravenously or intramuscularly with PBS, peptide, 500 nm SSHEL, or 500 nm SSHEL EncPep and boosted via the same inoculation route 2 weeks later. After 1 week, blood sample was collected, CD3e/CD8a double positive cytotoxic T cells were isolated by flow cytometry and analyzed for their ability to bind to MHC-I/SIINFEKL. (B) Representative flow cytometry plot of isolated CD3e/CD8a positive from mice inoculated intramuscularly with 500 nm SSHEL EncPep that recognize the SIINFEKL tetramer. (C) Percentage of CD3e/CD8a positive cytotoxic T cells that recognize SIINFEKL when presented in MHC-I harvested from mice injected (left) intravenously or (right) intramuscularly with (black) PBS, (pink) free peptide, (blue) SSHEL, or (green) SSHEL EncPep .

Article Snippet: Next, samples were stained using 10 μL of MHC Class I Murine iTAg Tetramer/APC – H-2 Kb OVA (SIINFEKL) (MBL International Corporation, Cat No. TB-5001-2) for 30 min at room temperature, protected from light, then washed according to manufacturer protocol.

Techniques: Injection, Isolation, Flow Cytometry