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Pre-immature dendritic cells (PIDC) pulsed with HPV16 E6 or E7 peptide are capable of eliciting specific immune response in patients with advanced cervical cancer.

Rahma OE, Herrin VE, Ibrahim RA, Toubaji A, Bernstein S, Dakheel O, Steinberg SM, Abu Eid R, Mkrtichyan M, Berzofsky JA, Khleif SN - J Transl Med (2014)

Bottom Line: HPV16 E6 was found to be homologous to HPV18 E6 in both vivo and vitro.There were no RECIST responses in any patient.The majority of toxicities were grade I and II.

View Article: PubMed Central - PubMed

Affiliation: Cancer Vaccine Branch, CCR, NCI, 10 Center Drive, Bethesda 20892, MD, USA. skhleif@gru.edu.

ABSTRACT

Background: The protein products of the early genes E6 and E7 in high-risk HPV types 16 and 18 have been implicated in the oncogenic capability of these viruses. Therefore, these peptides represent attractive vaccine therapy targets.

Methods: Thirty-two patients with advanced cervical cancer (HPV16 or 18 positive) were treated with HPV16 E6 (18-26) (Arm A) or HPV16 E7 (12-20) peptide (Arm B) pulsed on PBMCs in order to illicit immune response against the relevant peptide on both arms. These PBMCs were cultured for a short time (48 hours only) and in the presence of GM- CSF, accordingly, they were identified as "Pre-Immature Dentritic Cells".

Results: 51Cr release assay and ELISPOT demonstrated evidence of specific immune response against the relevant peptide in 10/16 (63%) evaluable patients in arm A and 7/12 (58%) in arm B. HPV16 E6 was found to be homologous to HPV18 E6 in both vivo and vitro. The median overall survival (OS) and progression free survival (PFS) for the full cohort was 10.0 and 3.5 months, respectively. There were no RECIST responses in any patient. The majority of toxicities were grade I and II.

Conclusions: We demonstrated the feasibility and ability of Pre-Immature Dentritic Cells pulsed with HPV16 E6 (18-26) or HPV16 E7 (12-20) to induce a specific immune response against the relevant peptide despite the advanced disease of the cervical cancer patients treated on this trial. We believe that this observation deserves further investigations.

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FACS analysis showing the phenotype of PIDCs used in the vaccine preparation. A) After a two day incubation in the presence of GM-CSF, and pulsating with HPV E7 peptide for two hours, CD14 positive cells persistently expressed CD14 at a slightly lower level. B) CD14 positive cells expressed a higher level of CD80 following incubation with GM-CSF and HPV E7 peptide. C) CD14 positive cells expressed a higher level of CD86 following incubation with GM-CSF and HPV E7 peptide. D) CD14 positive cells expressed a higher level of CD1a following incubation with GM-CSF and HPV E7 peptide. E) CD14 positive cells expressed a higher level of the Mannose receptor (CD206) following incubation with GM-CSF and HPV E7 peptide. F) HLA-DR was up-regulated in CD14 positive cells following incubation with GM-CSF and HPV E7 peptide.
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Fig1: FACS analysis showing the phenotype of PIDCs used in the vaccine preparation. A) After a two day incubation in the presence of GM-CSF, and pulsating with HPV E7 peptide for two hours, CD14 positive cells persistently expressed CD14 at a slightly lower level. B) CD14 positive cells expressed a higher level of CD80 following incubation with GM-CSF and HPV E7 peptide. C) CD14 positive cells expressed a higher level of CD86 following incubation with GM-CSF and HPV E7 peptide. D) CD14 positive cells expressed a higher level of CD1a following incubation with GM-CSF and HPV E7 peptide. E) CD14 positive cells expressed a higher level of the Mannose receptor (CD206) following incubation with GM-CSF and HPV E7 peptide. F) HLA-DR was up-regulated in CD14 positive cells following incubation with GM-CSF and HPV E7 peptide.

Mentions: FACS analysis was done on the PIDCs before vaccine administration to document their phenotype. The data was further confirmed using PBMCs from healthy donors. Prior to the GM-CSF incubation; the PIDCs were all monocytes expressing CD14 and they did not express any of the antigen presenting cell surface markers. After two days in culture in the presence of GM-CSF, and pulsing for two hours with the HPV16 E6 or E7 peptide, in spite of the persistent expression of CD14 at slightly lower levels (Figure 1A), CD14 positive cells (both from PIDCs and PBMCs) were found to express antigen presenting cell associated surface markers (CD80, CD86, CD1a and Mannose receptor (CD206)) (Figure 1B-E). Furthermore, there was up regulation in HLA-DR (Figure 1F).Figure 1


Pre-immature dendritic cells (PIDC) pulsed with HPV16 E6 or E7 peptide are capable of eliciting specific immune response in patients with advanced cervical cancer.

Rahma OE, Herrin VE, Ibrahim RA, Toubaji A, Bernstein S, Dakheel O, Steinberg SM, Abu Eid R, Mkrtichyan M, Berzofsky JA, Khleif SN - J Transl Med (2014)

FACS analysis showing the phenotype of PIDCs used in the vaccine preparation. A) After a two day incubation in the presence of GM-CSF, and pulsating with HPV E7 peptide for two hours, CD14 positive cells persistently expressed CD14 at a slightly lower level. B) CD14 positive cells expressed a higher level of CD80 following incubation with GM-CSF and HPV E7 peptide. C) CD14 positive cells expressed a higher level of CD86 following incubation with GM-CSF and HPV E7 peptide. D) CD14 positive cells expressed a higher level of CD1a following incubation with GM-CSF and HPV E7 peptide. E) CD14 positive cells expressed a higher level of the Mannose receptor (CD206) following incubation with GM-CSF and HPV E7 peptide. F) HLA-DR was up-regulated in CD14 positive cells following incubation with GM-CSF and HPV E7 peptide.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4269078&req=5

Fig1: FACS analysis showing the phenotype of PIDCs used in the vaccine preparation. A) After a two day incubation in the presence of GM-CSF, and pulsating with HPV E7 peptide for two hours, CD14 positive cells persistently expressed CD14 at a slightly lower level. B) CD14 positive cells expressed a higher level of CD80 following incubation with GM-CSF and HPV E7 peptide. C) CD14 positive cells expressed a higher level of CD86 following incubation with GM-CSF and HPV E7 peptide. D) CD14 positive cells expressed a higher level of CD1a following incubation with GM-CSF and HPV E7 peptide. E) CD14 positive cells expressed a higher level of the Mannose receptor (CD206) following incubation with GM-CSF and HPV E7 peptide. F) HLA-DR was up-regulated in CD14 positive cells following incubation with GM-CSF and HPV E7 peptide.
Mentions: FACS analysis was done on the PIDCs before vaccine administration to document their phenotype. The data was further confirmed using PBMCs from healthy donors. Prior to the GM-CSF incubation; the PIDCs were all monocytes expressing CD14 and they did not express any of the antigen presenting cell surface markers. After two days in culture in the presence of GM-CSF, and pulsing for two hours with the HPV16 E6 or E7 peptide, in spite of the persistent expression of CD14 at slightly lower levels (Figure 1A), CD14 positive cells (both from PIDCs and PBMCs) were found to express antigen presenting cell associated surface markers (CD80, CD86, CD1a and Mannose receptor (CD206)) (Figure 1B-E). Furthermore, there was up regulation in HLA-DR (Figure 1F).Figure 1

Bottom Line: HPV16 E6 was found to be homologous to HPV18 E6 in both vivo and vitro.There were no RECIST responses in any patient.The majority of toxicities were grade I and II.

View Article: PubMed Central - PubMed

Affiliation: Cancer Vaccine Branch, CCR, NCI, 10 Center Drive, Bethesda 20892, MD, USA. skhleif@gru.edu.

ABSTRACT

Background: The protein products of the early genes E6 and E7 in high-risk HPV types 16 and 18 have been implicated in the oncogenic capability of these viruses. Therefore, these peptides represent attractive vaccine therapy targets.

Methods: Thirty-two patients with advanced cervical cancer (HPV16 or 18 positive) were treated with HPV16 E6 (18-26) (Arm A) or HPV16 E7 (12-20) peptide (Arm B) pulsed on PBMCs in order to illicit immune response against the relevant peptide on both arms. These PBMCs were cultured for a short time (48 hours only) and in the presence of GM- CSF, accordingly, they were identified as "Pre-Immature Dentritic Cells".

Results: 51Cr release assay and ELISPOT demonstrated evidence of specific immune response against the relevant peptide in 10/16 (63%) evaluable patients in arm A and 7/12 (58%) in arm B. HPV16 E6 was found to be homologous to HPV18 E6 in both vivo and vitro. The median overall survival (OS) and progression free survival (PFS) for the full cohort was 10.0 and 3.5 months, respectively. There were no RECIST responses in any patient. The majority of toxicities were grade I and II.

Conclusions: We demonstrated the feasibility and ability of Pre-Immature Dentritic Cells pulsed with HPV16 E6 (18-26) or HPV16 E7 (12-20) to induce a specific immune response against the relevant peptide despite the advanced disease of the cervical cancer patients treated on this trial. We believe that this observation deserves further investigations.

Show MeSH
Related in: MedlinePlus