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Lectin Griffithsin Through Tobacco Mosaic Virus to Inhibit Human Immunodeficiency Virus
(HIV)/Asquired Immuno Deficiency Syndrom (AIDS). 2017 November 2.
Published in final edited form as:
Lectin Griffithsin Through Tobacco Mosaic Virus to Inhibit Human Immunodeficiency Virus (HIV) / Asquired
Immuno Deficiency Syndrom (AIDS). 2017 November 2nd

Lectin Griffithsin Through Tobacco Mosaic Virus To Inhibit

Human Immunodeficiency Virus (HIV) / Acquired Immuno
Deficiency Syndrom (AIDS)
1
Suci Hidayatur Rohmah
1
Student of Faculty of Dentistry, Universitas Jember.

Abstract
HIV (Human Immunodeficiency Virus) is a virus that attacks the immune system and weakens
the body's ability to fight infections and diseases. Prevention can use a lectin called griffithsin
(GRFT). Recombinant GRFT can be obtained by plasmid method. GRFT at Nicotiana
Benthamiana (N. Benthamiana) uses a vector based on tobacco mosaic virus (TMV). The
TMV-based expression vector system contains the GRFT amino acid sequence used to
generate transmitted RNA transcripts. Transmitted transcripts were inoculated on N.
Benthamiana which was then purified using rTMV. Griffithsin inhibits HIV by reducing viral
toxicity and stability.

Keywords
Griffithsin; TMV; HIV

INTRODUCTION
Human Immunodeficiency Virus (HIV) or Acquired Immuno Deficiency Syndrome (AIDS)
is one of the health problems in Indonesia. The number of HIV / AIDS cases continues to
increase from year to year. This raises concern not only for Indonesia but for all the nations
of the world. HIV / AIDS has had an impact in the socio-economic field, such as the impact
on demographics, the impact on health care systems and the impacts of the national
economy. According to Ministry of Health Surveillance Report, from January to June 2011
there were 2,352 new HIV / AIDS cases with a total of 26,483 people. The majority of
cases of HIV / AIDS are from young adult group, ie from the age group 20-29 years, with
46.4 percent of the total number of patients. This is certainly very worrying because the
younger generation is the next generation of the nation. Various things have been done to
prevent the spread of HIV / AIDS. Prevention is done by various forms of education.
Researchers on the other hand have also discovered a substance that can treat and inhibit
HIV / AIDS, namely griffithsin (Purnomo, 2013).

METHODS
Two methodological procedures were taken to conduct this review. First, a review of the
cancer survivorship literature was conducted in order to identify and summarize.

Lectin Griffithsin Through Tobacco Mosaic Virus to Inhibit Human Immunodeficiency Virus (HIV) / Asquired Immuno
Deficiency Syndrom (AIDS); 2017 November 2.
 RESULTS
Literature Review
The researchers succeeded in extracting bioassays from red Griffithsia algae species originating
from waters in New Zealand with extracts of Griffithsin lectins (GRFT). This lectin has anti-viral
function after testing in the laboratory as a primary HIV isolate in the picomolar range. This
GRFT is considered a potential microbicide candidate to prevent sexual transmission of HIV /
AIDS. This is an important discovery in the medical world but these algae are hard to find and
the numbers are limited in the open seas so researchers need innovation to produce large amounts
of griffithsin quickly and cheaper (Kouokam, 2016).

The recombinant Griffithsin used in this study was produced at Nicotiana benthamiana which is
the type of tobacco most vulnerable to Tobacco Mozaic Virus (TMV). TMV that usually infects
tobacco is used as a tool to produce GRFT. TMV rod-shaped, with a width of 18 nanometers and
a length of 200-300 nanometers. In his body is injected a red algae gene that produces griffithsin.
TMV virus that already contains red algae DNA will produce GRFT (Kouokam, 2016).

The female reproductive tract is covered by mucus, a gel glycoprotein composed of 95% water, 1
to 2% mucin fiber, and constituent elements (including salts, DNA, lactic acid, enzymes, and
other proteins). The virus must travel through the mucosal barrier to reach the epithelial or
subepithelial cells to be able to infect. Cerviksovaskinal mucus is a congenital protection against
infection through mucin-glycoproteins that interact with mucoadhesives physically to reduce
pathogenic flux to the epithelium. However, mucin's natural affinity interactions can actually
provide a basis for viral attachment and entry into cells by decreasing pathogenflux in
cervicalovaginal mucus (Grooms, 2016).

Griffithsin is a broad spectrum as an antiviral lectin that inhibits the entry and maturation of the
virus through the glycerol oligomannosa binder groups in viral envelope glycoproteins. GRFT
recombinant proteins can be obtained from Escherichia coli and Nicotiana benthamiana. GRFT
recombinant results in E. coli (ECGRFT) were 819 mg per liter culture medium but 33% protein
accumulated as inclusion bodies while GRFT in N. Benthamiana using vaccine based on tobacco
mosaic virus (TMV) yielded production up to 1 gram of GRFT per kg of fresh leaf weight.
(Vamvaka, 2016).

Griffirthsin is expressed in N. benthamiana by a plasmid method. Briefly, the TMV-based vector

expression system contains the GRFT amino acid sequence used to produce transcript RNA
transmitted. The infectious transcript was inoculated on N. benthamiana (~ 24 days post sowing)
with soft abrasion. GRFT extract is purified by using an AKTA cleaning tool with an XK16
column packed with 10mL Capto ™ MMC resin (GE Life Sciences). After overnight incubation
Semi-pure extract is filtered through a 0.2 μm filter and loaded into columns at 5 mL / min
(Fuqua, 2015).

The researchers then attempted to optimize the original purification process for mass production
of GRFT from the laminar network N. benthamiana using the rTMV expression vector. The new
process can drastically improve GRFT recovery that generates 60-90% of the total GRFT
expressed earlier. GRFT expression in N. benthamiana using rTMV can be performed using
transmitted nontransmitted RNAs from plasmids or the use of first viral particles isolated from
RNA-infected plants. Original DNA plasmids containing a subsequent dual subgenomic
promoter that transmitted TMVU1 sequentially in the upper portion of the rear-coat protein
promoter shown in the sequence shown by N. benthamiana with GRFT cloning codon of rTMV
vector, designed using heterologous mild mild mosaic virus mosaic (U5) 30 sequences, cistron
protein coat and 30 UTR (Figure 1). Infected plants are examined for contact with TMV.

Lectin Griffithsin Through Tobacco Mosaic Virus to Inhibit Human Immunodeficiency Virus (HIV) / Asquired Immuno
Deficiency Syndrom (AIDS); 2017 November 2.
 Symptoms at domains of envelope proteins from neutralizing Abs and to facilitate infections of the langerhans
harvest time and dendritic cells (O'Keefe, 2009).
are
determined From biological-based entry inhibitors, the red algae griffithsin protein (GRFT) has the greatest
by qualifying activity as a potent inhibitor and anti-HIV, with an average EC50 of 40 μM. GRFT targets
metrics terminal mannose residues on high mannose oligosaccharides and has a total of 6 carbohydrate
requiring binding sites per homodimer that can be accounted for as unmatched potential. Inactivation of
90% of the HIV-1 by GRFT occurs immediately in contact with the virus. The GRFT antiviral potential is
plants augmented by cellular toxicity deficiencies, exceptional environmental stability (resistance to
inoculated various pH and temperature near the boiling point), and stability of cervical leprosy
for symptoms cervixovaskinal lymph nodes GRFT is the ideal active ingredient for topical HIV microbicides
(Figures 1a (O'Keefe, 2009).
and b). For
CONCLUSION
qualification
of the Griffithsin is a lectin that can inhibit HIV by reducing the toxicity and stability of the virus.
inoculum, the Recombinant Griffithsin can be obtained through Nicotiana Benthamiana using a vector based on
infected plant tobacco mosaic virus (TMV). GRFT is expressed in N. benthamiana by using plasmid method.
should
produce Supplementary Material
excess GRFT
Refer to Web version on Google Schoolar for supplementary material.
of 200 mg /
kg of
biomass Acknowledgments
(Fuqua, I am deeply indebted to my lecturer, Dr. drg. Banun Kusumawardani, M. Kes for warm support,
2015). inspiration and thoughtful guidance

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HIV uses
glycans to
Figures
protect the
important

Lectin Griffithsin Through Tobacco Mosaic Virus to Inhibit Human Immunodeficiency Virus (HIV) / Asquired Immuno
Deficiency Syndrom (AIDS); 2017 November 2.
 particles of rTMV. Plants not expressing GRFT or infected with rTMV showed no visible symptoms TMV
infection.

Fig
ure
1. Figure 1. (b) Plants expressing GRFT using rTMV showed leaf powder, growth retardation and leaf
(a). deformation. The inset image is a close-up of visible growth deformities, with curling of the leaves and
Grif significantly reducing the size of the infected leaf.
fith
sin
exp
ress
es
infe
ctio
us
plan
ts
and

Figure 1. (c) purified rTMV virion Used for GRFT expression imaged with a transmission electron
microscope under 46K magnification after uranyl acetate staining. Whole full-length virions can be
confirmed with a minimal number of partial particles.

Lectin Griffithsin Through Tobacco Mosaic Virus to Inhibit Human Immunodeficiency Virus (HIV) / Asquired Immuno
Deficiency Syndrom (AIDS); 2017 November 2.