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Combination of conserved recombinant proteins (NP & 3M2e) formulated with Alum protected BALB/c mice against influenza A/PR8/H1N1 virus challenge

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Abstract

Purpose

Influenza is one of the most important agents of pandemic outbreak causing substantial morbidity and mortality. Vaccination strategies of influenza must be adapted annually due to constant antigenic changes in various strains. Therefore, the present study was conducted to evaluate protective immunity of the conserved influenza proteins.

Methods

For this purpose, three tandem repeats of M2e (3M2e) and NP were separately expressed in E. coli and were purified using column chromatography. Female Balb/c mice were injected intradermally with a combination of the purified 3M2e and NP alone or formulated with Alum (AlOH3) adjuvant in three doses. The mice were challenged by intranasal administration of H1N1 (A/PR/8/34) 2 weeks after the last vaccination.

Results

The results demonstrated that recombinant NP and M2e proteins are immunogenic and could efficiently elicit immune responses in mice compared to non-immunized mice. The combination of 3M2e and NP supplemented with Alum stimulated both NP and M2e-specific antibodies, which were higher than those stimulated by each single antigen plus Alum. In addition, the secretion of IFN-γ and IL-4 as well as the induction of lymphocyte proliferation in mice received the mixture of these proteins with Alum was considerably higher than other groups. Moreover, the highest survival rate (86%) with the least body weight change was observed in the mice immunized with 3M2e and NP supplemented with Alum followed by the mice received NP supplemented with Alum (71%).

Conclusion

Accordingly, this regimen can be considered as an attractive candidate for global vaccination against influenza.

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Data availability

All data are available in case of need.

Abbreviations

3M2e:

Three tandem repeats of M2e

M2e:

M2 extracellular domain

HA:

Hemagglutinin

NA:

Neuraminidase

UIV:

Universal influenza vaccines

NP:

Nucleoprotein

IAV:

Influenza A virus

rNP:

Recombinant nucleoprotein

Th2:

T Helper 2

IgG1:

Immunoglobulin G1

IgE:

Immunoglobulin E

SDS:

Acrylamide and sodium dodecyl sulphate

IPTG:

Isopropyl β-d-1-thiogalactopyranoside

TMB:

Tetramethylbenzidine

3-(4,5-dimethyl tetrazolyl-2) 2,5 diphenyl:

MTT salt

HRP:

Horseradish peroxidase

ELISA:

Enzyme-linked immunosorbent assay

LEW:

Lysis-equilibration-wash

PBS:

Phosphate buffer saline

DMSO:

Dimethyl sulfoxide

SI:

Stimulate index

IFN-γ:

Interferon-γ

IL-4:

Interleukin 4

LD90 :

Lethal Dose 90

APCs:

Antigen-presenting cells

HSP70:

Heat shock proteins

CTL:

Cytotoxic T lymphocyte

LB Broth:

Luria-Bertani Broth

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Acknowledgements

This study was performed in the Department of Influenza and Other Respiratory Viruses, Pasteur Institute of Iran and was also financially supported by the Pasteur Institute of Iran (Grant Number: 759). Hereby, the authors would like to show their gratitude to all the staff working in Pasteur Institute of Iran for their valuable assistance and cooperation.

Funding

This work was supported by the Pasteur Institute of Iran (Grant Number 759).

Author information

Authors and Affiliations

  1. Department of Microbiology and Microbial Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, District 1, Daneshjou Boulevard, 1983969411, Tehran, Iran

    Mehrnaz Forqani & Seyed Masoud Hosseini

  2. Department of Influenza and Other Respiratory Viruses, Pasteur Institute of Iran, No.69, 12th Farvardin Street, Pasteur Avenue, 1316943551, Tehran, Iran

    Mehrnaz Forqani, Behrokh Farahmand, Maryam Saleh, Hadiseh Shokouhi, Ali Torabi & Fatemeh Fotouhi

Authors
  1. Mehrnaz Forqani
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  2. Seyed Masoud Hosseini
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  3. Behrokh Farahmand
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  4. Maryam Saleh
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  5. Hadiseh Shokouhi
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  6. Ali Torabi
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  7. Fatemeh Fotouhi
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Contributions

(MS): Study concept and design; Development of the protocol; Acquisition, analysis and interpretation of data; Draft of the manuscript, wrote and prepared the manuscript; and Critical revision of the manuscript for important intellectual content. (JN): Study concept and design; Supervision; Administrative; Technical and material support. (BF): Study concept and design; Supervision; Administrative; Technical, and material support. (FF): Study concept and design; Development of the original idea and the protocol; Administrative; Technical, and material support; Supervision; and Interpretation of data.

Corresponding author

Correspondence to Fatemeh Fotouhi.

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Authors declare that they have no conflict of interests.

Ethical approval

All the animal experiments were approved and performed based on the guidelines of the Ethical Committee of the Pasteur Institute of Iran, Tehran, Iran (IR.PII.REC.1395.82).

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Forqani, M., Hosseini, S.M., Farahmand, B. et al. Combination of conserved recombinant proteins (NP & 3M2e) formulated with Alum protected BALB/c mice against influenza A/PR8/H1N1 virus challenge. Biotechnol Lett 43, 2137–2147 (2021). https://doi.org/10.1007/s10529-021-03174-2

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