Journal: NPJ Vaccines

Article Title: Overcoming immunogenicity issues of HIV p24 antigen by the use of innovative nanostructured lipid carriers as delivery systems: evidences in mice and non-human primates

doi: 10.1038/s41541-018-0086-0

Figure Lengend Snippet: Immunogenic properties of NLC formulations. a Antigen presentation by primary DC to anti-OVA T lymphocytes. Secretion of IL-13 and IFN-γ by T lymphocytes after 4 days of co-culture with DC previously stimulated with formulations combining OVA and CpG. Bars indicate the standard error of the mean calculated on three measurements. b Humoral response in mice as measured based on serum anti-p24 and anti-OVA antibody levels and c cell-mediated response as determined based on the secretion of IFN-γ by T lymphocytes from the spleen of immunized mice after in vitro antigen stimulation. Mice were immunized with both p24 and OVA formulations (5 µg p24 and 10 µg OVA) co-administered with 10 µg CpG. Each point represents an individual mouse, horizontal bars represent the mean for the group and vertical bars indicate the standard error of the mean. Data were compared between groups using a 1-way ANOVA test followed by Fisher’s protected least significant difference test. *** p < 0.001

Article Snippet: Thiol groups were introduced on p24 (p24–SH) by derivatization of 1 mg p24 (PX’Therapeutics) with 10 µg of 2-iminothiolane (Thermo Scientific).

Techniques: Co-Culture Assay, In Vitro

Journal: NPJ Vaccines

Article Title: Overcoming immunogenicity issues of HIV p24 antigen by the use of innovative nanostructured lipid carriers as delivery systems: evidences in mice and non-human primates

doi: 10.1038/s41541-018-0086-0

Figure Lengend Snippet: a Humoral response in mice following immunization with 5 µg of p24 and 10 µg of OVA in combination with 10 µg of CpG formulations, as measured based on total anti-p24 and anti-OVA antibody levels in muse serum. b IgG1, IgG2a, and IgA isotypes were also analyzed and are expressed as the highest serum dilution giving an absorbance of more than three times the background. Each point represents an individual mouse, horizontal bars represent the mean for the group and vertical bars indicate the standard error of the mean. Data were compared between groups using a 1-way ANOVA test followed by a Fisher’s protected least significant difference test. ***p < 0.001; **p < 0.01; *p < 0.05

Article Snippet: Thiol groups were introduced on p24 (p24–SH) by derivatization of 1 mg p24 (PX’Therapeutics) with 10 µg of 2-iminothiolane (Thermo Scientific).

Techniques:

Journal: NPJ Vaccines

Article Title: Overcoming immunogenicity issues of HIV p24 antigen by the use of innovative nanostructured lipid carriers as delivery systems: evidences in mice and non-human primates

doi: 10.1038/s41541-018-0086-0

Figure Lengend Snippet: T-cell response after immunizing mice with 5 µg of p24 and 10 µg of OVA formulations in combination with 10 µg of CpG formulations, as measured based on IFN-γ, TNF-α, and IL-2 secretion by splenocytes after 72 h of antigen stimulation. Each point represents an individual mouse, horizontal bars represent the mean for the group and vertical bars indicate the standard error of the mean. Data were compared between groups using a 1-way ANOVA test followed by a Fisher’s protected least significant difference test. *** p < 0.001; ** p < 0.01; * p < 0.05

Article Snippet: Thiol groups were introduced on p24 (p24–SH) by derivatization of 1 mg p24 (PX’Therapeutics) with 10 µg of 2-iminothiolane (Thermo Scientific).

Techniques:

Journal: NPJ Vaccines

Article Title: Overcoming immunogenicity issues of HIV p24 antigen by the use of innovative nanostructured lipid carriers as delivery systems: evidences in mice and non-human primates

doi: 10.1038/s41541-018-0086-0

Figure Lengend Snippet: Antigen-specific CD4 and CD8 T-cell responses after immunization of mice with 5 µg of p24 and 10 µg of OVA co-administered with 10 µg of CpG formulations, as measured by intracellular staining (ICS). Splenocytes were stimulated with antigen for 24 h and then CD4 and CD8 T lymphocytes were stained intracellularly for IFN-γ and analyzed by flow cytometry. Each point represents an individual mouse, horizontal bars represent the mean for the group and vertical bars indicate the standard error of the mean. Data were compared between groups using a 1-way ANOVA test followed by Fisher’s protected least significant difference test. ***p < 0.001; ** p < 0.01; * p < 0.05

Article Snippet: Thiol groups were introduced on p24 (p24–SH) by derivatization of 1 mg p24 (PX’Therapeutics) with 10 µg of 2-iminothiolane (Thermo Scientific).

Techniques: Staining, Flow Cytometry

Journal: NPJ Vaccines

Article Title: Overcoming immunogenicity issues of HIV p24 antigen by the use of innovative nanostructured lipid carriers as delivery systems: evidences in mice and non-human primates

doi: 10.1038/s41541-018-0086-0

Figure Lengend Snippet: Immune responses in macaques ( N = 4 per group) following four intradermal immunizations with NLC formulations containing the equivalent of 64 µg p24 and 253 µg CpG. a Specific antibodies and b IFN-γ-producing T lymphocytes. Individual macaques are represented by circular, triangular, square or diamond-shaped markers. Dashes represent the mean for each group, and bars indicate the standard error of the mean. Arrows below the X-axis indicate immunizations. Data were compared between groups base on a 2-way ANOVA test followed by Holm–Sidak method. ***p < 0.001; **p < 0.01; * p < 0.05 between NLC–p24+ NLC+ /CpG group and others

Article Snippet: Thiol groups were introduced on p24 (p24–SH) by derivatization of 1 mg p24 (PX’Therapeutics) with 10 µg of 2-iminothiolane (Thermo Scientific).

Techniques:

Journal: NPJ Vaccines

Article Title: Overcoming immunogenicity issues of HIV p24 antigen by the use of innovative nanostructured lipid carriers as delivery systems: evidences in mice and non-human primates

doi: 10.1038/s41541-018-0086-0

Figure Lengend Snippet: Graphical representation of NLC interactions with immune cells in lymph nodes. Within lymph nodes, DC would be activated by the capture of both NLC–p24 and NLC+ /CpG or by inflammatory cytokines produced by neighboring DC having captured NLC+ /CpG. Depending on p24 processing, antigenic peptides will be loaded on MHC-II or MHC-I for presentation to CD4 or CD8 T lymphocytes, respectively. Specific antibodies will be produced by B cells stimulated by Th2 cells, while CD8 T lymphocytes will differentiate into CTL following stimulation by Th1 cells. DC dendritic cell, TL T lymphocyte, BL B lymphocyte. Image credit: for the dendritic cell image, Donald Bliss, National Library of Medicine (source: Sriram Subramaniam, National Cancer Institute, 2010); for the T and B lymphocyte images, Blausen.com staff (2014), WikiJournal of Medicine 1 (2)

Article Snippet: Thiol groups were introduced on p24 (p24–SH) by derivatization of 1 mg p24 (PX’Therapeutics) with 10 µg of 2-iminothiolane (Thermo Scientific).

Techniques: Produced

Journal: NPJ Vaccines

Article Title: Overcoming immunogenicity issues of HIV p24 antigen by the use of innovative nanostructured lipid carriers as delivery systems: evidences in mice and non-human primates

doi: 10.1038/s41541-018-0086-0

Figure Lengend Snippet: Physicochemical properties of antigen-grafted NLC

Article Snippet: Thiol groups were introduced on p24 (p24–SH) by derivatization of 1 mg p24 (PX’Therapeutics) with 10 µg of 2-iminothiolane (Thermo Scientific).

Techniques: