Encephalitis (JE)
Clinical features Diagnosis
The clinical disease can be divided into The institute has a strong infrastructure
General Since 1972, JE has spread to newer
three stages - a prodromal febrile stage, and the following facilities are
JE is an important mosquito-borne viral areas and epidemics / outbreaks have
been reported from West Bengal, Uttar an acute encephalitic stage marked by established, standardized and regularly
disease and one of the leading causes of used for providing diagnosis.
Pradesh, Assam, Manipur, Bihar, CNS involvement and continuing fever
viral encephalitis and neurological
A n d h r a P r a d e s h , Po n d i c h e r r y, and a late stage marked by either
infections in Asia. Although severely Virus isolations & propagation
Karnataka, Goa and recently from recovery or persistence of symptoms of
under-reported, 50,000 cases are Ÿ Tissue culture
Kerala and Maharashtra. The disease in irreversible neuron injury leading to
annually recorded throughout Asia, with Ÿ Infant mouse inoculation
southern India affects children below 15 transient / permanent sequelae.
15,000 deaths (5-35% case fatality rate) Ÿ Mosquito inoculation
years, while in north India all age groups Subclinical infections are relatively
and a 75% JE-related disability rate
are affected. In most of the epidemics, high; sub clinical to clinical infection
(767,000 DALYs, WHO, 2002). Antigen detection
the incidence has been higher in males ratio being 125-500:1.
Ÿ Antigen capture ELISA
The disease was first recognized in India than in females (M : F=1.2 -1.5:1). Ÿ Immunofluorescence test
in 1955, when cases of encephalitis The virus
from North Arcot district of Tamil Nadu An enveloped virus, belongs to the Serological tests
Distribution family Flaviviridae. The virus is spherical
and neighboring districts of Andhra Based on endemicity, epidemics, Ÿ Haemagglutination Inhibition (HI )
Pradesh, admitted to Christian Medical and has diameter of 40-50 nm. Ÿ ELISA (IgM, IgG)
isolations of virus from patients and
College Hospital, Vellore, were vectors and general serological surveys, Ÿ Neutralization
serologically diagnosed as Japanese the virus has been shown to be widely
Encephalitis. JE virus was isolated from prevalent in most parts of south central, Genome detection
wild caught mosquitoes in the same northern and northeast states of India. Ÿ PCR
year, followed by isolations from patients Apparently, part of Maharashtra and
from the same area in 1958. JE states like Gujarat, Rajasthan and
continues to be endemic in these states. Madhya Pradesh are free from JE.
315bp
108bp
Genome
Single stranded positive sense RNA of Diagnostic PCR using 2 different regions
~11000 nucleotides. The virus has
three structural and 7 non-structural The institute has developed highly
(NS) proteins that are translated as a reliable IgM capture ELISA.
single ORF and co - or post- (MAC-ELISA) for rapid diagnosis.
translationally cleaved. The virus NIV supplies a limited number of
replicates exclusively in cytoplasm. The kits on commercial basis.
outer protein (envelope) has domains
responsible for cell attachment,
haemagglutination and neutralization.
Global prevalence
5’- C PrM E NS1 NS3 NS5 -3’
(Source: WHO)
Prevalence in India
15
Situations under which
mosquitogenic conditions occur are
! Water accumulation and paddy
cultivation during monsoon season
! Paddy cultivation associated with
irrigation systems during non-
monsoon seasons eg. Mandya district,
Karnataka
Mosquito collection at dusk
! Water logging due to flooding of rivers
eg. Brahmaputra river basin, Assam Vector mosquitoes rest outdoors and are
AMPLIFYING
predominantly zoophilic in nature.
HOST ! Changing of agricultural practices Only <2% of the females feed on
MAINTENANCE such as dryland wheat cultivation to human blood. Therefore, very high
HOST
paddy cultivation using ground vector density is a prerequisite for active
and/or canal water eg. Uttar Pradesh. transmission to humans. Strong
TRANSOVARIAL
TRANSMISSION association has been demonstrated
between JE cases and vector densities /
longevity of mosquitoes.
16
17
Immunology
19
Indian strains belong to cluster III and IV
4
93 NHGN- Ns1 (145)
97 NAKAYAMA
NHGN- MEM(9.5)
98
SARAWAK. MALASIA 3
50% of subjects after two doses (0 and
No survivo
SRI LANKA TLD-MEM(102)
82 IV
97
SAJOON VIETHAM
KAMIYAMA JAPAN
Vaccine development 2 NS1-TLD(175)
PBS (7)
7-17 days) and in ~60% of subjects
31 VELLORE Challenge protocol 1 Vaccine(21) after the third dose given after 13-17
JaOAr. JAPAN
Challenge of mice with intraperitoneal months.
III O
39 60
80
BANKURA
GOA
virus followed by intracerebral 1 3 5 7 9 11 13 15 17 19 21
BEIJING-P3 inoculation of 1% starch has been studied Days post challance 2. Trial on 113 school children in
98
Dh20, NEPAL
II as challenge protocol to determine the Protection of mice immunized with chimeric Andhra Pradesh
INDONESIA
88
99 Hk8256, TAIWAN efficacy of vaccine. This takes into peptide from lethal challenge of JE virus Seventy-three percent of the subjects
BEIJING
account the role of CMI response in sero-converted after 2 doses (day 0 and
74 60 Sa14, CHINA
95
Sa14-2-8 VACCINE I
addition to antibody response in 14). A booster dose administered one
93 Sa14-5-3 VACCINE protection. year later resulted in sero-conversion in
92 Sa14-14-2 VACCINE
KPP, THAILAND
Predicted structure of 88% of the subjects. However, it was
Attenuated vaccine Chimeric peptide noted that 79% had shown significant
MAUR, SINGAPUR
WEST NILE Partial attenuation of JE virus was drop in anti-JEV antibodies in pre-
l Sequencing carried out at NIV
obtained by passages in chick embryo
DNA vaccine
booster samples.
culture. However, no stable attenuated The precursor M-envelope (truncated)
Sequencing the envelope gene of and Ns1 genes were cloned into a
strain could be obtained. Both these studies indicated that
neutralization escape mutants mammalian expression vector to
minimum of three doses of vaccine are
revealed the following develop candidate DNA vaccine. The
One attenuated ts mutant obtained from necessary.
! Substitutions of G by W (Vellore) and E persistently infected cells showed loss of recombinant plasmids expressed
by to K (Bankura) are the sites involved virulence at passage level of 20 by the i.p. antigen in transfected cells. Mice
in loss of neutralization and loss of immunized with recombinant plasmid Prevention and control
route. This mutant could protect mice
virulence. against challenge with 2.5 logs of virus showed the presence of neutralizing strategies
! Changing the substrate for growing and thus has the potential to be antibodies. Data collected on population dynamics
virus from mouse brain to tissue developed as an attenuated vaccine. of vectors have helped in evolving
culture and sequencing the envelope mosquito control programs. Insecticide
gene revealed that mutations at Tissue culture vaccine susceptibility tests were also performed
position 84, 184, 211, 219 and 490 A chick embryo culture derived on field collected mosquitoes. Control
are responsible for change in tropism inactivated unpurified vaccine candidate programs are implemented by State
Expression of JE virus Governments under the guidance and
from mouse to tissue culture (PS cells). was protective in mice. protein in COS-7 cells
supervision of National Malaria
! All the mutations are in the domain I
Peptide vaccine Eradication Program.
and II, involved in dimerization of the Vaccine trials
monomers, important for cell Using chimeric peptides incorporating
sequences of T helper and B cell epitopes, NIV has conducted two JE vaccine trials Central Research Institute, Kasauli,
interaction and virus entry. using Nakayama mouse brain derived
N.antibody induction and partial manufactures mouse brain derived
protection of mice from lethal challenge killed vaccine. Nakayama strain killed vaccine.
with JE virus has been demonstrated. Production is limited and endemic states
Studies on increasing the protective 1. Trial on 120 laboratory volunteers occasionally vaccinate children. There is
ability of these chimeric peptides using (adults) strong need to produce better and cost
antigen delivery systems and adjuvant The sero-conversion to neutralizing effective vaccine for mass
are planned. These chimeric peptide antibody response was detected in only immunization.
epitopes are being used for development
Three dimensional structure of JE virus envelop of polytope DNA vaccine. JE is a priority disease for NIV. The main thrust areas
proteitn showing locations of peptide epitopes include rapid diagnosis, molecular epidemiology, immunology,
20 vaccine development and vaccine trials.