Leishmania major heat shock protein 70 (HSP70) is not protective in murine models of cutaneous leishmaniasis and stimulates strong humoral responses in cutaneous and visceral leishmaniasis patients

Abstract

Heat shock proteins (HSP) are highly conserved molecules that play important roles in protein folding, assembly of protein complexes and translocation of proteins across cellular compartments, as well as in several immunological processes. In this study, we first immunized susceptible BALB/c and resistant C57BL/6 mice with the complete open-reading frame of Leishmania HSP-70 (pcDNA-HSP70) and boosted mice with rHSP-70 (amino acid 221-604 cloned in pQE-HSP70 and referred to as rHSP70) mixed with Montanide 720. When we evaluated the effects of HSP70 in both mouse strains, we found that the entire fragment (amino acids 221-604) and rCT-HSP70 (amino acids 491-604 cloned in pQE-CT), but not rNT-HSP70 (amino acids 221-291 cloned pQE-NT), contained the highest immunogenicity. However, after infectious challenge with Leishmania major, no efficient protective responses were observed in either mouse strain. The humoral immune responses against the different truncated forms of HSP70 suggested a mixed TH1/TH2 response in vivo. We then assessed infected susceptible and resistant mice for lymphoproliferative and cytokine responses against the truncated forms of HSP70. At 9-week post-infection, we observed no differences in those responses between vaccinated and control mice. Next, we initiated comparative studies in human patient samples, finding no significant proliferation against all three truncated forms of HSP70 in the cellular immune responses of 16 cured cutaneous leishmaniasis patients and 5 normal individuals. Sera from active cutaneous and visceral leishmaniasis patients, however, were reactive to all three forms of HSP70. This study demonstrates the potential of HSP70 in stimulating humoral responses in humans and mice and indicates there is a need to further explore and examine the value of this important molecule in the control of leishmaniasis.