T cell response to oncoprotein E7 of Human Papillomavirus genotype 16 (HPV 16) delivered by Salmonella typhi strain Ty21a (S. typhi Ty21a) in mice

Order of Publishing in Issue: 
2
Volume :6
Issue :4
October, 2012
Page No: 
398-406
Authors: 
Ponnanna NM[1], Rajan Sriraman[1], Ravi Ranjan Verma [1], Swapna E[1], K. Balaji [1], K.Sathish[1], V. Maroudam [1], R.K. Sisodiya[1], M Lakshmi Narasu [2] and VA Srinivasan[1]
Address: 
[1] Research & Development Centre Indian Immunologicals Limited Rakshapuram, Gachibowli, Hyderabad 500032, Andhra Pradesh, India
Address: 
[2] Center for Biotechnology, Jawaharlal Nehru Technological University, Hyderabad

Abstract

Immunotherapy is intensely being considered for either a stand-alone or supplemental treatment of HPV associated precancers and cancers. The aim of the study was to enquire whether S. typhi Ty21a engineered to express the HPV 16 E7 (Ty21a-E7) elicits T cell response to the oncoprotein. The codon optimized synthetic HPV16 E7 gene was cloned in an expression vector under a constitutive promoter (Ptac). Female mice of strain C57BL/6 were immunized intra nasally with Ty21a-E7. The cell mediated immune response was evaluated employing the IFN?-Elispot assay. Splenocytes were stimulated with the affinity purified recombinant proteins, either GST-E7 or GST expressed in E. coli BL21. The HPV 16 E7 specific IFN secreting- spot forming cells (SFCs) obtained on stimulation with purified GST-E7 in the Elispot assay was 240 (±6.92) cells/million splenocytes, significantly higher (p< 0.01) than the number for splenocytes stimulated with GST (67.3 ±26.40 cells/million). The antigen specific T cells were found to be predominantly of the CD4+ T cell type. The results show E7 specific CMI response in the mouse model of study. To our knowledge this is the first study reporting CMI response in mice to an HPV oncoprotein delivered by S. typhi Ty21a- the live-attenuated, oral, typhoid vaccine strain.

Keywords: 
HPV 16 E7, Elispot, T cell response, CD4+ T cells, Ty21a-E7.
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