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Membrane-bound HSP70-engineered myeloma cell-derived exosomes stimulate more efficient CD8(+) CTL- and NK-mediated antitumour immunity than exosomes released from heat-shocked tumour cells expressing cytoplasmic HSP70.

Xie Y, Bai O, Zhang H, Yuan J, Zong S, Chibbar R, Slattery K, Qureshi M, Wei Y, Deng Y, Xiang J - J. Cell. Mol. Med. (2010)

Bottom Line: We found that EXO(HSP) were able to more efficiently stimulate maturation of DCs with up-regulation of Ia(b) , CD40, CD80 and inflammatory cytokines than EXO(HS) after overnight incubation of immature bone-marrow-derived DCs (5 × 10⁶ cells) with EXO (100 μg), respectively.We demonstrate that EXO(HSP) are able to stimulate type 1 CD4(+) helper T (Th1) cell responses, and more efficient P1A-specific CD8(+) cytotoxic T lymphocyte (CTL) responses and antitumour immunity than EXO(HS) .In addition, we further elucidate that EXO(HSP) -stimulated antitumour immunity is mediated by both P1A-specific CD8(+) CTL and non-P1A-specific natural killer (NK) responses.

View Article: PubMed Central - PubMed

Affiliation: Research Unit, Division of Health Research, Saskatchewan Cancer Agency, Department of Oncology, University of Saskatchewan, Saskatoon, Canada.

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Animal studies. (A) BALB/c mice were s.c. vaccinated with EXOneo, EXOHS and EXOHSP or PBS. 6 days after immunization, the immunized mice were s.c. inoculated with J558 tumour cells. Animal mortality and tumour growth were monitored daily for up to 60 days. *, P < 0.05 versus cohorts in EXOHSP group (log-rank test). (B) Kinetic tetramer staining assay. 2 to 8 days after immunization of mice with EXOHSP, the tail blood samples were taken from the immunized mice, stained with PE-H-2Ld/P1A peptide tetramer and FITC anti-CD8 Ab and analysed by flow cytometry. The value in each panel represents the percentage of tetramer+ CD8+ T cells versus the total blood CD8+ T-cell population. (C) Kinetic study of NK activity. 2 to 8 days after immunization of mice with EXOHSP, the mouse spleen T cells were tested for NK killing activity to J558 tumour cells. BALB/c mice with treatment of anti-CD8 or anti-NK Ab were s.c. vaccinated with either (D) EXOneo or (E) EXOHS (F) EXOHSP. 6 days after immunization, the immunized mice were s.c. inoculated with J558 tumour cells. Animal mortality and tumour growth were monitored daily for up to 6 weeks. One representative experiment of three is shown.
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fig05: Animal studies. (A) BALB/c mice were s.c. vaccinated with EXOneo, EXOHS and EXOHSP or PBS. 6 days after immunization, the immunized mice were s.c. inoculated with J558 tumour cells. Animal mortality and tumour growth were monitored daily for up to 60 days. *, P < 0.05 versus cohorts in EXOHSP group (log-rank test). (B) Kinetic tetramer staining assay. 2 to 8 days after immunization of mice with EXOHSP, the tail blood samples were taken from the immunized mice, stained with PE-H-2Ld/P1A peptide tetramer and FITC anti-CD8 Ab and analysed by flow cytometry. The value in each panel represents the percentage of tetramer+ CD8+ T cells versus the total blood CD8+ T-cell population. (C) Kinetic study of NK activity. 2 to 8 days after immunization of mice with EXOHSP, the mouse spleen T cells were tested for NK killing activity to J558 tumour cells. BALB/c mice with treatment of anti-CD8 or anti-NK Ab were s.c. vaccinated with either (D) EXOneo or (E) EXOHS (F) EXOHSP. 6 days after immunization, the immunized mice were s.c. inoculated with J558 tumour cells. Animal mortality and tumour growth were monitored daily for up to 6 weeks. One representative experiment of three is shown.

Mentions: To investigate the antitumour immunity derived from EXOHSP vaccination, BALB/c mice were s.c. immunized with EXOHSP, EXOneo, EXOHS and PBS, respectively. 6 days after the immunization, the immunized mice were s.c. challenged with J558 tumour cells. As shown in Fig. 5(A), all the mice injected with PBS died of tumour within 14 days after tumour cell challenge. EXOHS and EXOneo vaccine protected four of eight (50%) and two of eight (25%) mice from tumour growth, respectively, and the rest of four or six tumour-bearing mice had significantly delayed tumour growth compared to the control group of mice treated with PBS (P < 0.05). However, EXOHSP immunization protected all eight of eight (100%) mice from tumour growth, indicating that EXOHSP expressing membrane-bound HSP70 can also induce more efficient antitumour immunity than EXOHS expressing cytoplasmic HSP70 and the control EXOneo without HSP70 expression.


Membrane-bound HSP70-engineered myeloma cell-derived exosomes stimulate more efficient CD8(+) CTL- and NK-mediated antitumour immunity than exosomes released from heat-shocked tumour cells expressing cytoplasmic HSP70.

Xie Y, Bai O, Zhang H, Yuan J, Zong S, Chibbar R, Slattery K, Qureshi M, Wei Y, Deng Y, Xiang J - J. Cell. Mol. Med. (2010)

Animal studies. (A) BALB/c mice were s.c. vaccinated with EXOneo, EXOHS and EXOHSP or PBS. 6 days after immunization, the immunized mice were s.c. inoculated with J558 tumour cells. Animal mortality and tumour growth were monitored daily for up to 60 days. *, P < 0.05 versus cohorts in EXOHSP group (log-rank test). (B) Kinetic tetramer staining assay. 2 to 8 days after immunization of mice with EXOHSP, the tail blood samples were taken from the immunized mice, stained with PE-H-2Ld/P1A peptide tetramer and FITC anti-CD8 Ab and analysed by flow cytometry. The value in each panel represents the percentage of tetramer+ CD8+ T cells versus the total blood CD8+ T-cell population. (C) Kinetic study of NK activity. 2 to 8 days after immunization of mice with EXOHSP, the mouse spleen T cells were tested for NK killing activity to J558 tumour cells. BALB/c mice with treatment of anti-CD8 or anti-NK Ab were s.c. vaccinated with either (D) EXOneo or (E) EXOHS (F) EXOHSP. 6 days after immunization, the immunized mice were s.c. inoculated with J558 tumour cells. Animal mortality and tumour growth were monitored daily for up to 6 weeks. One representative experiment of three is shown.
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fig05: Animal studies. (A) BALB/c mice were s.c. vaccinated with EXOneo, EXOHS and EXOHSP or PBS. 6 days after immunization, the immunized mice were s.c. inoculated with J558 tumour cells. Animal mortality and tumour growth were monitored daily for up to 60 days. *, P < 0.05 versus cohorts in EXOHSP group (log-rank test). (B) Kinetic tetramer staining assay. 2 to 8 days after immunization of mice with EXOHSP, the tail blood samples were taken from the immunized mice, stained with PE-H-2Ld/P1A peptide tetramer and FITC anti-CD8 Ab and analysed by flow cytometry. The value in each panel represents the percentage of tetramer+ CD8+ T cells versus the total blood CD8+ T-cell population. (C) Kinetic study of NK activity. 2 to 8 days after immunization of mice with EXOHSP, the mouse spleen T cells were tested for NK killing activity to J558 tumour cells. BALB/c mice with treatment of anti-CD8 or anti-NK Ab were s.c. vaccinated with either (D) EXOneo or (E) EXOHS (F) EXOHSP. 6 days after immunization, the immunized mice were s.c. inoculated with J558 tumour cells. Animal mortality and tumour growth were monitored daily for up to 6 weeks. One representative experiment of three is shown.
Mentions: To investigate the antitumour immunity derived from EXOHSP vaccination, BALB/c mice were s.c. immunized with EXOHSP, EXOneo, EXOHS and PBS, respectively. 6 days after the immunization, the immunized mice were s.c. challenged with J558 tumour cells. As shown in Fig. 5(A), all the mice injected with PBS died of tumour within 14 days after tumour cell challenge. EXOHS and EXOneo vaccine protected four of eight (50%) and two of eight (25%) mice from tumour growth, respectively, and the rest of four or six tumour-bearing mice had significantly delayed tumour growth compared to the control group of mice treated with PBS (P < 0.05). However, EXOHSP immunization protected all eight of eight (100%) mice from tumour growth, indicating that EXOHSP expressing membrane-bound HSP70 can also induce more efficient antitumour immunity than EXOHS expressing cytoplasmic HSP70 and the control EXOneo without HSP70 expression.

Bottom Line: We found that EXO(HSP) were able to more efficiently stimulate maturation of DCs with up-regulation of Ia(b) , CD40, CD80 and inflammatory cytokines than EXO(HS) after overnight incubation of immature bone-marrow-derived DCs (5 × 10⁶ cells) with EXO (100 μg), respectively.We demonstrate that EXO(HSP) are able to stimulate type 1 CD4(+) helper T (Th1) cell responses, and more efficient P1A-specific CD8(+) cytotoxic T lymphocyte (CTL) responses and antitumour immunity than EXO(HS) .In addition, we further elucidate that EXO(HSP) -stimulated antitumour immunity is mediated by both P1A-specific CD8(+) CTL and non-P1A-specific natural killer (NK) responses.

View Article: PubMed Central - PubMed

Affiliation: Research Unit, Division of Health Research, Saskatchewan Cancer Agency, Department of Oncology, University of Saskatchewan, Saskatoon, Canada.

Show MeSH
Related in: MedlinePlus