A Review on Nanovaccines, its Need for Diseases Control, Advantages, Limitation and

Commercialization.
Abstract
Vaccination often refers to the prevention of diseases, (i.e.) it prevents future malfunctioning in
our body. The main causes of most of the malfunctions are the pathogens/microbes which get
entry in our body directly or indirectly. Thus, a war begins between these antigens and our cell
system. The immune system in our body helps in defeating the antigens to an extent, but certain
antigens are capable enough to overcome the immune defense of our body. These are commonly
auto-immune and also genetic diseases inherited from parents to offspring. One big query that
arises is do we have a solution that can monitor the activities of cells, tissues, and organs. So, we
need an artificial mechanism that, like blood cells circulates in our bloodstream. Nanoparticle
based vaccines have proven prodigious prophylaxis of various human and animal associated
infectious and non-infectious diseases. Nanovaccines have the potential to induce both antibody-
mediated and cell-mediated immunity and can render long-term immunogenic memory. With
such properties, nano-vaccines have shown high potential for the prevention of infectious
diseases such as acquired immunodeficiency syndrome (AIDS), malaria, tuberculosis, influenza,
and cancer.
This review will discuss the development of nano-vaccines and their administration into the body
via different routes. As well as the applications, advantages, limitations and the types of
nanoparticles used in the preparation of vaccines used for the prevention of infectious diseases
(having bacterial, viral or other parasitic origin) and non-infectious diseases (like cancer, auto-
immune diseases) using nano-vaccinology.
Introduction
“Prevention is better than cure”. It has been the mankind's goal since earlier ages to get
protection against life-threatening ailment. The idea of vaccination flourished when Edward
Jenner discovered that cowpox protects individuals from smallpox and ever since then the
medical field became interested in improving the immunity against diseases some which can be
fatal [1]
However, it is not always such a simple task to develop a vaccine and so many studies were
started to gain detailed knowledge about the immune system recognition to antigens resulting in
the synthesis of many vaccines with antibody and cell mediated mechanisms of protection.[2]
Most of the difficulties appear when we try to develop vaccines against viruses like HIV, HCV
etc. which are known to be much variable and where the commonly followed approach of
vaccine formulation ‘isolate, inactivate, inject’ is not helpful. For the past many years, high
throughput technology has proven to be necessary in order to get proper knowledge about the
complex biological systems and their immune systems and finally utilize these technologies in
the development of new vaccines. [3]
And so, in order to step forward with improving immunogenicity, the use of nanotechnology in
the field of vaccinology appeared and thus the term nanovaccinology was given importance.
 Nano materials are defined as structures in between the size of 1–100 nm dimension, have started
to be generally used for developing the vaccines is mainly to help in prophylactic approaches and
manyare approved to use, as well as in therapeutic approaches which is used in cancer to a great
extent, in addition to treatment of other diseases such as Alzheimer's, hypertension etc.[4]
Brief insights and way forward for commercialization of nano-vaccines in clinical settings have
been summarized in the entire research innovation.
1. Classification of Vaccines
Nanovaccines can be divided into diverse classes based on different parameters including: shape,
source, sizes, features and structural constriction.
Natural: Bacterial spores, Viruses like particles, Bacteriophages, Exosomes.
Synthetic:Proteosomes, Nano-beads, Liposomes, Virosomes.

2. Need of Nanovaccines for Disease Control

The nanoparticles with efficient surface properties are more suitable candidates for stimulating
the immune system and eliciting better immunological response. On the basis of low-
functionalized plasmid DNA, high lability to degrading enzymes, lacking the smart size and
hydrophobic nature the first and second-generation vaccines differ from nano-vaccines.

Examples and clinical status of nanocarriers-based vaccines.

Targeted diseases Nanocarrier Exploited antigen Clinical status
Influenza T7 phage VLP HA, M2e Preclinical
Influenza Human ferritin cage M2e Preclinical
Influenza α-helix self- assembling peptide M2e/CFA+IFA Preclinical
nanoparticles
Influenza α-helix self- assembling peptide Helix C, M2e/flagellin, PADRE Preclinical
nanoparticles
Influenza Liposomes H1N1 Split virus Phase 1
Malaria Self-assembling protein nanoparticle FMP014 Preclinical
(SAPN)
Malaria β-sheet fibers (NANP)3 Preclinical
Malaria α-helix fibers PbCSP Preclinical
Malaria Liposomes RTS,S Preclinical
Malaria Iron oxide nanoparticles Merozoite surface protein Phase ½
HIV α-helix self- assembling peptide 2F5, 4E10/IFA Preclinical
nanoparticles
HIV Gold nanoparticles HIV-1 Env plasmid Preclinical
Tuberculosis β-sheet fibers ESAT6, TB10.4, Ag85B/Pam2Cys Preclinical
Tuberculosis Self-assembling peptide nanofibers Mtb-specific CD8+ or CD4+ T cell Preclinical
epitopes
Pneumonia Qβ phage Tetra saccharide/NKT cell adjuvant Preclinical
Pneumonia Polyanhydride nanoparticle Pneumococcal surface protein A Preclinical
(PspA)
Respiratory syncytial virus Liposome Envelope (E) protein Preclinical
Hepatitis B Chitosan nanoparticles Recombinant Hepatitis B surface Preclinical
antigen (rHBsAg)
Dengue Liposomes Envelope (E) protein Preclinical
Meningitidis Liposomes Outer membrane proteins and Phase 1
deacetylated lipo-oligosaccharide
Tetanus Gold nanoparticles Tetanus toxoid bulk from Clostridium Preclinical
tetani
Foot and mouth disease Gold nanoparticles FMDV peptide Preclinical
Melanoma Hepatitis B VLP Melanoma peptides Preclinical
Cancer Metal organic framework nanoparticle Cytomembrane of fused DCs and Preclinical
Cancer Cells
Breast cancer Liposomes dHER2 protein Phase 1
Lung cancer Liposomes BLP25 Phase 3

i) Nanovaccines for Viruses

HIV is the sixth leading cause of death across the globe. Its infection causes systemic depletion
of CD4+ T cells, hence compromising the activity of the body’s immune system and the
development of AIDS. Immune functions can be preserved in the early stages of HIV by using
antiretroviral therapy (ART) [5]
ii) Nanovaccines for cancer prevention
The cancer vaccines have failed to make a mark clinically comparative to other immunotherapies
like T cell therapy and checkpoint blockade yet. The only therapeutic cancer vaccine, Provenge
(Sipuleucel-T), approved by US FDA, has shown a moderate increase in clinical efficacy in the
defense against prostate cancer.
iii) Nanovaccines for tuberculosis
Being the deadliest infectious disease, Tuberculosis is the greatest cause of death across the
globe. 10% of infected individuals develop the disease within one or two years of infection and
the rest develop the disease in later stages of life when immune functions are common. BCG
(Bacille Calmette-Guerin) is a commonly accepted vaccine against TB (Tuberculosis), but its
protective efficacy is limited by age due to the absence of consensus genomic loci that are
present in several virulent strains of Mtb. It induces short term and variable immune responses
from 0% to 80% [6]
iv) Nanovaccines for malaria
Malaria affects almost 200 million people per year, with half a million deaths across the globe.
Due to its multistage life cycle, fighting malaria is challenging. Many efforts have been made to
design vaccines against the pre-erythrocytic and blood stage of malaria. The defense responses
generated against sexual stage antigens of malaria reduces its transmission and lessens the
disease burden. Pfs25 is one such malaria transmission-blocking vaccine antigen, but its
immunogenicity is limited in humans.[7]

3. Administration of Nanovaccines
Nasal Route
Nasal sprays formulated exhibited advantages like its small size, the ability of being easily dis
posed and delivering limited number of sprays.
Oral Route
In gene therapy, the possible idea of administration of DNA vaccine with the aid of polymer-
based nanoparticles via oral route was first proposed by by Bhavsar and Aniji. In light of this, the
attempts of using oral DNA vaccination against diseases are initiated.
Subcutaneous Route
Several studies have been performed to evaluate the transport of particles like liposomes by
injecting subcutaneously to the lymph nodes. A nanovaccine developed against leishmaniasis,
which contain recombinant Leishmania superoxide dis mutase loaded onto chitosan
nanoparticles has been examined and administered subcutaneously and resulted in an increase in
the immunogenicity using this approach. [8]
Intramuscular Route
In certain cases, the antigen may not be able to reach the Langerhans cells of the epidermis and
thus intramuscular can be an alternate route used. The use of this approach has been reported in
formulating vaccine against influenza. Intramuscular administration of virus-like particles for
vaccination of H5N1 has reported to be effective[9]
Intradermal Route.
Drugs and genes can be administered for the purpose of immunization by intradermal route
which is one of the most common routes used[10]
Topical Route
Vaccination can be delivered via skin and skin patches usually used to deliver genetic materials
as plasmid DNA. These have been utilized in increasing the levels of hepatitis B surface antigen
as well as cytokines which improves immunization.
Ocular Route
The eye being at the surface an exposed to the external environment, is susceptible to get
infected by antigens and infectious materials and so a vaccination through the ocular mucosa is
acceptable. Vaccination for influenza H1N1 by eye drops induced antibody response against
respiratory infection.
Vaginal Route
Administration through vaginal route serves benefits like high permeability, large surface area,
bypassing first pass metabolism and a route which is rich with blood supply making it a
reasonable approach especially with it being a route with the potential of being infected with
many pathogens.

4. Commercialization (Marketed Products)

 As discussed earlier, the nanoparticles used in the production of vaccines have been put into
actual use generating wide range of Nanovaccines having many applications. Some are still in
the primary stages of clinical trials while others have reached the market and are being utilized
by the patients.
No. Name Type of NPs Company Use Clinical Reference
Stages
1 DepoDur™ liposome Sky PharmaInc. Pain management FDA [11]
approved
2 Inflexal® Virosomes Crucell, Berna Biotech Subunit influenza vaccine FDA [11]
approved
3 FluMist® N-trimethylchitosan-mono-N- Medimmune Quadrivalent live attenuated FDA [12]
carboxymethyl influenza vaccines approved
chitosannanocomplexes
4 Oncept® Naked plasmid Merial Protection against dog Licensed [13]
melanoma by USDA
5 Provenge™ Dendritic cells Valeant Prostate cancer Phase I/II [14]
Pharmaceuticals clinical
trials
6 LifeTide® Therapeutic plasmids VGX Animal Health Reducing perinatal morbidity Approved
and mortality by
APVMA
7 Apex-IHN® DNA vaccine Novartis Animal Health Prevents infectious Licensed
haematopoietic necrosis by CFIA
8 DP6-001 Multiple-antigenpolyvalent CytRx Corporation HIV vaccine Phase I
DNA vaccine clinical
trial
9 ZYC101 Microencapsulated DNA Eisai Pharmaceuticals Treat dysplasia by HPV Phase I [14]
vaccine clinical
trial
10 Vaxfectin® Liposomes Vical DNA immunization for Phase I [15]
H5N1 clinical
trial

Advantages
Nanovaccines have many advantages nowadays, from their wide size range by their enhanced
bioavailability. They can easily enter through cellular mechanism called endocytosis
[3]Nanoemulsions vaccines acquire high affinity for the lymphatic system, therefore they can be
easily gathered in the lymph nodes regions. Such novel nanovaccination systems can be used for
influenza prophylaxis, cancer immunotherapy and other applications which still under studies
and researches[16] nanotechnology in vaccine provide the opportunity of enhancing the safety
and its stability via enhancing both cellular and humoral immune systems. Interestingly,
vaccination using nanopatch technology is pain free, needle free and cost-effective as well as
provide enhanced antigens bio availability and antigens site-specific delivery. [17] Refrigeration
was not required for this since nanoemulsion was effective for 30 days at 26°C, and for 45 days
at 42°C.
Disadvantages
Behavior of thenanoparticle invaccine invivo makesa big challenge
forthescientisttodevelopanewnano-vaccine. The liposomal delivery systems [3]show a great
drawback in the formation of nano-vaccine which tends to aggregate during storage.[3] Nano-
vaccine technology considers a complicated procedure for production new vaccines which
requires a serious of machines and tools to use, so this makes the nano-vaccine method
production very costly. Due to over a prolonged cleared period, with nano-vaccines, this may
induce toxicity.
Conclusion
The discovery and evolution of vaccines formulations from the classical vaccines in the medical
field have been considered as a masterstroke in human history. Nanotechnology played an
important role in the evolution of nano vaccines; which offers properties that may render high
effectiveness and safety in vaccines.
‘Nano-vaccinology’ is the science dealing with nanoscale particles with huge potential. The
laboratory as well as the clinical scale processing of nano-vaccines can provide an eco-friendly,
more immunogenic, sustained and stabilized releasing novel approach against infectious and
non-infectious diseases. Integrity, in terms of desirable surface properties during production and
storage of nano-vaccines in field conditions should be addressed while manufacturing nano-
vaccines commercially. The nano-vaccines have provided boundless hopes in efficiently
preventing pathogenic, cancerous and non-infectious diseases in immune-tolerant people. More
research studies in collaboration with commercial industries can result in quick
commercialization of nano-vaccines.
Many of the recently conducted clinical trials demonstrated the potential of nanovaccines
intreating as well as preventing many infectious diseases. However, most of the formulations are
still in their testing stages and the number of the commercialized products is relatively low but
with the encouraging results of the conducted clinical trials, are revolutionary improvement in
the field of immunization is highly predicted.

References
1. Li, S., et al., Systems biological approaches to measure and understand vaccine immunity
in humans. Seminars in Immunology, 2013. 25(3): p. 209-218.
2. Oberg, A.L., et al., Systems biology approaches to new vaccine development. Current
Opinion in Immunology, 2011. 23(3): p. 436-443.
3. Zhao, L., et al., Nanoparticle vaccines. Vaccine, 2014. 32(3): p. 327-337.
4. Pulendran, B., S. Li, and H.I. Nakaya, Systems Vaccinology. Immunity, 2010. 33(4): p.
516-529.
5. Aikins, M.E., J. Bazzill, and J.J. Moon, Vaccine nanoparticles for protection against HIV
infection. Nanomedicine, 2017. 12(6): p. 673-682.
6. Ansari, M.A., et al., RD Antigen Based Nanovaccine Imparts Long Term Protection by
Inducing Memory Response against Experimental Murine Tuberculosis. PLOS ONE,
2011. 6(8): p. e22889.
7. Kumar, R., et al., Nanovaccines for malaria using Plasmodium falciparum antigen Pfs25
attached gold nanoparticles. Vaccine, 2015. 33(39): p. 5064-5071.
8. Sekhon, B.S. and V. Saluja, Nanovaccines-an overview. Int J Pharm Front Res, 2011.
1(1): p. 101-109.
9. Nandedkar, T., Nanotechnology a path to nanovaccine. Int. J. Pharma Bio Sci, 2012.
3(2): p. 290-292.
10. Nandedkar, T.D., Nanovaccines: recent developments in vaccination. Journal of
Biosciences, 2009. 34(6): p. 995-1003.
11. Bulbake, U., et al., Liposomal Formulations in Clinical Use: An Updated Review.
Pharmaceutics, 2017. 9(2): p. 12.
12. Bernocchi, B., R. Carpentier, and D. Betbeder, Nasal nanovaccines. International Journal
of Pharmaceutics, 2017. 530(1): p. 128-138.
13. Glikin, G.C. and L.M.E. Finocchiaro, Clinical trials of immunogene therapy for
spontaneous tumors in companion animals. The Scientific World Journal, 2014. 2014.
14. Racz, R., et al., DNAVaxDB: the first web-based DNA vaccine database and its data
analysis. BMC Bioinformatics, 2014. 15(4): p. S2.
15. Ferraro, B., et al., Clinical Applications of DNA Vaccines: Current Progress. Clinical
Infectious Diseases, 2011. 53(3): p. 296-302.
16. Li, X., et al., Nanomaterials in the application of tumor vaccines: advantages and
disadvantages. OncoTargets and therapy, 2013: p. 629-634.
17. Zaman, M., M.F. Good, and I. Toth, Nanovaccines and their mode of action. Methods,
2013. 60(3): p. 226-231.