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Published on: July 17, 2014
Plasmodium falciparum Heat Shock Protein 70 Lacks Immune Modulatory Activity
Ofentse Jacob Pooe1, Gabriele Köllisch2, Holger Heine3
1Department of Biochemistry, Westville Campus, University of KwaZulu-Natal, Private Bag X54001, Durban 4000. South Africa.
Background:
Heat shock protein 70 (Hsp70) family are conserved molecules that constitute a major part of the cell's protein folding machinery. The role of Hsp70s of parasitic origin in host cell immune modulation has remained contentious. This is largely due to the fact that several studies implicating Hsp70 in immune modulation rely on the use of recombinant protein derived from bacteria which is often fraught with lipopolysaccharide (LPS) contamination. For this reason, there is need to clarify the role of parasite Hsp70 in modulating host immune cells.
Objective:
The current study sought to investigate the role of Plasmodium falciparum Hsp70 (PfHsp70) in immune modulation.
Method:
We expressed recombinant PfHsp70 using three bacterial expression hosts: E. coli XL1 Blue, E. coli ClearColi BL21 and Brevibacillus choshinensis, respectively. We further investigated the immunostimulatory capability of PfHsp70 by monitoring cytokine production by murine immune cells cultured in the presence of the protein.
Results:
Recombinant PfHsp70 produced using E. coli XL1 Blue expression host induced IL6 and IL8 cytokines. On the other hand, PfHsp70 produced in E. coli ClearColi and B. choshinensis expression systems was associated with no detectable traces of LPS and exhibited no immunomodulatory activity.
Conclusion:
Our findings demonstrate that PfHsp70 does not possess immunomodulatory function. Furthermore, our study further confirm E. coli ClearColi and B. choshinensis as appropriate expression systems for the production of LPS-free recombinant protein.
Insights
Parasitic Heat Shock Protein 70 (Hsp70) from Plasmodium falciparum does not modulate host immunity. Bacterial expression systems like E. coli ClearColi and B. choshinensis yield LPS-free Hsp70, crucial for accurate immune response studies.
Area of Science:
- Immunology
- Molecular Biology
- Parasitology
Background:
- Heat shock protein 70 (Hsp70) is vital for cellular protein folding.
- The immunomodulatory role of parasitic Hsp70s is debated, often due to bacterial lipopolysaccharide (LPS) contamination in recombinant proteins.
- Clarifying the function of parasite Hsp70 in host immune cells is essential.
Purpose of the Study:
- To investigate the immunomodulatory potential of Plasmodium falciparum Hsp70 (PfHsp70).
Main Methods:
- Recombinant PfHsp70 was expressed in three bacterial hosts: E. coli XL1 Blue, E. coli ClearColi BL21, and Brevibacillus choshinensis.
- The immunostimulatory activity of PfHsp70 was assessed by measuring cytokine production in murine immune cells exposed to the protein.
Main Results:
- PfHsp70 from E. coli XL1 Blue induced Interleukin-6 (IL6) and Interleukin-8 (IL8) production.
- PfHsp70 produced in E. coli ClearColi and B. choshinensis showed no detectable LPS and lacked immunomodulatory activity.
Conclusions:
- Plasmodium falciparum Hsp70 (PfHsp70) does not possess intrinsic immunomodulatory functions.
- E. coli ClearColi and B. choshinensis are suitable for producing LPS-free recombinant proteins for immunological research.
