Identification of the murine H-2Db and human HLA-A*0201 MHC class I-restricted HPV6 E7-specific cytotoxic T lymphocyte epitopes

Peng, Shiwen; Mattox, Austin K.; Best, Simon R.; Barbu, Anca M.; Burns, James A.; Akpeng, Belinda; Jeang, Jessica; Yang, Benjamin; Ishida, Eiichi; Hung, Chien Fu; Wu, T. C.; Pai, Sara I.

doi:10.1007/s00262-016-1793-x

Cited by 6 publications

(7 citation statements)

References 21 publications

(22 reference statements)

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“…HEK 293 expi cells were acquired from ATCC (Manassas, VA). The generation of 293 cells expressing the murine MHC class I molecule D b , 293-D b , or K b , 293-K b , was described previously (78). 293 expi, 293-Db, and 293-K b cells were cultured in Dulbecco's modified Eagle medium (DMEM) containing 2 mM glutamine, 1 mM sodium pyruvate, 100 IU ml 21 penicillin, 100 mg ml 21 streptomycin, and 10% FBS.…”

Section: Methodsmentioning

confidence: 99%

Development of DNA Vaccine Targeting E6 and E7 Proteins of Human Papillomavirus 16 (HPV16) and HPV18 for Immunotherapy in Combination with Recombinant Vaccinia Boost and PD-1 Antibody

Peng

Gaillard

Wang

et al. 2021

mBio

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Immunotherapy for cervical cancer should target high-risk human papillomavirus types 16 and 18, which cause 50% and 20% of cervical cancers, respectively. Here, we describe the construction and characterization of the pBI-11 DNA vaccine via the addition of codon-optimized human papillomavirus 18 (HPV18) E7 and HPV16 and 18 E6 genes to the HPV16 E7-targeted DNA vaccine pNGVL4a-SigE7(detox)HSP70 (DNA vaccine pBI-1). Codon optimization of the HPV16/18 E6/E7 genes in pBI-11 improved fusion protein expression compared to that in DNA vaccine pBI-10.1 that utilized the native viral sequences fused 3′ to a signal sequence and 5′ to the HSP70 gene of Mycobacterium tuberculosis. Intramuscular vaccination of mice with pBI-11 DNA better induced HPV antigen-specific CD8+ T cell immune responses than pBI-10.1 DNA. Furthermore, intramuscular vaccination with pBI-11 DNA generated stronger therapeutic responses for C57BL/6 mice bearing HPV16 E6/E7-expressing TC-1 tumors. The HPV16/18 antigen-specific T cell-mediated immune responses generated by pBI-11 DNA vaccination were further enhanced by boosting with tissue-antigen HPV vaccine (TA-HPV). Combination of the pBI-11 DNA and TA-HPV boost vaccination with PD-1 antibody blockade significantly improved the control of TC-1 tumors and extended the survival of the mice. Finally, repeat vaccination with clinical-grade pBI-11 with or without clinical-grade TA-HPV was well tolerated in vaccinated mice. These preclinical studies suggest that the pBI-11 DNA vaccine may be used with TA-HPV in a heterologous prime-boost strategy to enhance HPV 16/18 E6/E7-specific CD8+ T cell responses, either alone or in combination with immune checkpoint blockade, to control HPV16/18-associated tumors. Our data serve as an important foundation for future clinical translation. IMPORTANCE Persistent expression of high-risk human papillomavirus (HPV) E6 and E7 is an obligate driver for several human malignancies, including cervical cancer, wherein HPV16 and HPV18 are the most common types. PD-1 antibody immunotherapy helps a subset of cervical cancer patients, and its efficacy might be improved by combination with active vaccination against E6 and/or E7. For patients with HPV16+ cervical intraepithelial neoplasia grade 2/3 (CIN2/3), the precursor of cervical cancer, intramuscular vaccination with a DNA vaccine targeting HPV16 E7 and then a recombinant vaccinia virus expressing HPV16/18 E6-E7 fusion proteins (TA-HPV) was safe, and half of the patients cleared their lesions in a small study (NCT00788164). Here, we sought to improve upon this therapeutic approach by developing a new DNA vaccine that targets E6 and E7 of HPV16 and HPV18 for administration prior to a TA-HPV booster vaccination and for application against cervical cancer in combination with a PD-1-blocking antibody.

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Section: Methodsmentioning

confidence: 99%

Development of DNA Vaccine Targeting E6 and E7 Proteins of Human Papillomavirus 16 (HPV16) and HPV18 for Immunotherapy in Combination with Recombinant Vaccinia Boost and PD-1 Antibody

Peng

Gaillard

Wang

et al. 2021

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“…293-D b cell line has been previously described [ 14 ]. 293-D b cells were cultured in DMEM medium supplemented with 2 mM glutamine, 1 mM sodium pyruvate, 100 U/mL penicillin, 100 μg/mL streptomycin, and 10% fetal bovine serum.…”

Section: Methodsmentioning

confidence: 99%

Enhancing antitumor immunogenicity of HPV16-E7 DNA vaccine by fusing DNA encoding E7-antigenic peptide to DNA encoding capsid protein L1 of Bovine papillomavirus

et al. 2017

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BackgroundHuman papillomavirus (HPV) has been identified as the primary etiologic factor of cervical cancer, the fourth leading cause of cancer death in females worldwide. We have previously shown that coadministration of DNA encoding L1 capsid protein of Bovine papillomavirus (BPV) can enhance the antigen-specific immune response elicited by a therapeutic HPV16-E7 DNA vaccination. In this study, we sought to generate and evaluate the immunogenicity of a therapeutic HPV16-E7 DNA vaccine that encodes the fusion construct of HPV16-E7 and BPV-L1.ResultsWe generated a therapeutic HPV16-E7 DNA vaccine construct, pcDNA3-BPVL1-E7(49-57), encoding the fusion sequence of full-length BPVL1 protein and a murine E7 antigenic epitope, aa49-57. Transfecting 293-Db cells with pcDNA3-BPVL1-E7(49-57) demonstrated that this DNA construct can effectively lead to the presentation of E7 epitope for the activation of E7-specific CD8+ T cells in vitro. Intramuscular vaccination of pcDNA3-BPVL1-E7(49-57) with electroporation generated a stronger E7-specific CD8+ T cell-mediated immune response than coadministration of pcDNA3-BPVL1 and pcDNA3-E7(49-57) in C57BL/6 mice. Furthermore, we observed that the strong E7-specific CD8+ T cell response elicited by pcDNA3-BPVL1-E7(49-57) vaccination translated into potent protective and therapeutic antitumor effects in C57BL/6 mice against HPV16-E7 expressing TC-1 tumor cells. Finally, using antibody depletion experiment, we showed that the antitumor immune response generated by pcDNA3-BPVL1-E7(49-57) is CD8+ T cell dependent, and CD4+ T cell and NK cell independent.ConclusionTreatment with fusion construct of BPV-L1 and HPV16-E7 epitope can elicit effective E7-specific antitumor immune response in mice. Due to the potential ability of the fusion DNA construct to also trigger immune responses specific to the L1 protein, the current study serves to support future design of HPV DNA vaccines encoding fusion HPVL1-E6/E7 constructs for the generation of both T cell and B cell mediated immune responses against HPV infections and associated diseases.

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“…These data confirm previous findings using a DNA vaccine targeting HPV11 E6E7 that was not linked to CRT. 4 Although it is possible to induce an E7 response to vaccination by designing a vaccine containing only E7, 6 it appears that in the C57BL/6 background, E6 is the dominant immunologic antigen when both E6 and E7 proteins are expressed together. Vaccines designed for human use often include both antigens, to maximize the chances for antigen presentation, and there are many trials currently underway in HPV-associated diseases.…”

Section: Prophylactic Vaccines Against Low-and High-riskmentioning

confidence: 99%

“…In this report, it was demonstrated that E6 is the immunodominant peptide, but no specific immunologic response was seen against E7. One strategy to enhance immunologic responses and antigen presentation is linkage of the antigen of interest to calreticulin (CRT), and we then showed that linking CRT to HPV6‐E7 could generate an immune response against HPV6‐E7 not present in the non–CRT‐linked vaccine . However, the gold standard test for the efficacy of DNA vaccines is the ability to control growing tumors in vivo that express the target antigen.…”

Section: Introductionmentioning

confidence: 99%

“…One strategy to enhance immunologic responses and antigen presentation is linkage of the antigen of interest to calreticulin (CRT), 5 and we then showed that linking CRT to HPV6-E7 could generate an immune response against HPV6-E7 not present in the non-CRTlinked vaccine. 6 However, the gold standard test for the efficacy of DNA vaccines is the ability to control growing tumors in vivo that express the target antigen. So far, this has not been demonstrated with any tumor model This article was published online on July 17, 2017.…”

Section: Introductionmentioning

confidence: 99%

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Immunologic responses to a novel DNA vaccine targeting human papillomavirus‐11 E6E7

Ahn

Peng²,

Hung³

et al. 2017

The Laryngoscope

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Objective Recurrent respiratory papillomatosis (RRP) is a benign disease caused by human papillomavirus (HPV) types 6 and 11. Although a prophylactic vaccine against RRP is available, a therapeutic vaccine is needed to treat those already infected. The objective of our study was to design and test a DNA vaccine targeting HPV-11 proteins. Study Design Pre-clinical scientific investigation Methods A DNA vaccine encoding the HPV-11 E6 and E7 genes linked to calreticulin (CRT) was generated. Immunologic response to the HPV-11 CRT/E6E7 vaccine was measured by vaccinating C57BL/6 mice via electroporation and measuring CD8+ T cell responses from harvested splenocytes. A tumor cell line containing HPV11-E6E7 was created and the ability of novel DNA vaccine to control tumor growth was measured in vivo. Results Our vaccine generated a significant and specific CD8+ T cell response against the HPV11-E6aa41-70 peptide. The CD8+ T cell responses did not recognize E7 epitopes, indicating E6 immunodominance. CD8+ responses were augmented in the CRT-linked vaccine compared to a control non-CRT vaccine. The HPV-11 CRT/E6E7 vaccine was used to treat mice inoculated with a HPV-11 E6E7 expressing tumor cell line after temporary CD3 depletion to facilitate tumor growth. Vaccinated mice had a significantly lower tumor growth rate (p=0.029) and smaller tumor volumes compared to control mice indicating an augmented immunologic response in vaccinated mice. Conclusions A DNA vaccine targeting HPV-11 E6E7 generates a specific HPV-11 CD-8+ T cell response capable of reducing the growth of HPV-11 expressing tumors. DNA vaccines are a promising immunologic strategy for treating RRP. Level of Evidence N/A (Animal Study and basic research)

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Identification of the murine H-2Db and human HLA-A*0201 MHC class I-restricted HPV6 E7-specific cytotoxic T lymphocyte epitopes

Cited by 6 publications

References 21 publications

Development of DNA Vaccine Targeting E6 and E7 Proteins of Human Papillomavirus 16 (HPV16) and HPV18 for Immunotherapy in Combination with Recombinant Vaccinia Boost and PD-1 Antibody

Development of DNA Vaccine Targeting E6 and E7 Proteins of Human Papillomavirus 16 (HPV16) and HPV18 for Immunotherapy in Combination with Recombinant Vaccinia Boost and PD-1 Antibody

Enhancing antitumor immunogenicity of HPV16-E7 DNA vaccine by fusing DNA encoding E7-antigenic peptide to DNA encoding capsid protein L1 of Bovine papillomavirus

Immunologic responses to a novel DNA vaccine targeting human papillomavirus‐11 E6E7

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