DHF, HIV, POLIOMYELITIS, HERPES

SIMPLEKS 1 & 2, VARICELLA

ZOSTER, MORBILLI, MUMPS, RSV,
MOLLUSCUM CONTAGIOSUM, HPV

By Rizalinda Sjahril
Microbiology Department
FACULTY OF MEDICINE
HASANUDDIN UNIVERSITY
2018
11 VIRUSES THAT ARE DISCUSSED IN THIS
LECTURE:
1. Dengue Hemorrhagic Fever virus
2. Human Immunodeficiency virus
3. Poliomyelitis virus
4. Human Herpes Virus 1 (H. Simpleks oralis)
5. Human Herpes Virus 2 (H. Simpleks genitalis)
6. Human Herpes Virus 3 (Varicella-Zoster Virus)
7. Morbilli virus
8. Mumps
9. Respiratory Syncitial virus
10. Molluscum contagiosum virus
11. Human Papilloma Virus
 VIRUS AND DISEASE IT CAUSED
Virus Family Type virus Disease
Flavivirus Dengue Virus DF/DHF/DSS
Retroviridae Human Immunodeficiency AIDS
Virus
Herpesviridae Herpes simpleks 1 virus Herpes labialis
Herpes simpleks 2 virus Herpes genitalis
Varicella Zoster virus Chicken pox /shingles
Enteroviridae Poliovirus Polio
Paramyxovirus Morbillivirus Measles
Mumps virus Parotitis
Pneumovirus Bronchiolitis
Poxviridae Molluscumvirus Molluscum contagiosum
Papillomaviridae Human Papilloma Virus Warts (verruca vulgaris, etc)
Incidence/Epidemiology
Clinical symptoms
Virology/Taxonomy/Genomic
Virulence/Host Immune response
Laboratory Diagnosis
Prevention/Control

DENGUE HEMORRHAGIC FEVER VIRUS

 DISTRIBUTION OF DENV 2008

The contour lines indicate the potential geographical limits of the northern and southern hemispheres for
year-round survival of Ae. aegypti, the principal mosquito vector of DENVs
DENGUE VIRUS (DENV)
• Arbovirus : arthropod borne virus
• 4 serologically different Dengue viruses
• Clinical cases:

• Dengue fever (DF)  acute self-limiting fever

• Dengue hemorrhagic fever (DHF)
• Dengue shock syndrome (DSS).
• No effective DENV vaccine and antivirus. Trombositopenia and increased
vascular permeability
GLOBAL BURDEN OF DENV, 2010
a. Presence (red) and
absence of dengue
(green)

b. Possibility of dengue in
an area of 5 × 5 km.
High Probability (red);
Low Probability (green)

c. Annual infection in all

age compared to national
or sub-national proportion
in China.
Nature. 2013 Apr 25;496(7446):504-7
DENGUE INCUBATION PERIOD
• Extrinsic Incubation period
 = Virus replicates within female mosquito after biting an infected
human until the mosquito becomes infectious (8-12 days)
• Intrinsic Incubation period
 = After a mosquito bites a susceptibel human (one who has never
been infected before) until symptoms appear (4-7 days; may range
3-14 days)
• Period of communicability (human transmit virus to mosquito)
• Shortly before until the end of symptomatic viremia (4-5 days)
• Human never transmit to human; but transfusion related case is
possible
 CLINICAL SYMPTOMS OF DENGUE VIRUS
INFECTION
• Varies from asymptomatic to severe with bleeding and
shock (DHF/DSS).
• DHF is marked by plasma leakage and hemorrhagic
diathesis observed before defervescence, usually day
5 of fever.
• Death is due to hypotension and shock, often with
coagulation disorders and bleeding.
TAXONOMY/CLASSIFICATION
• Family: Flaviviridae
• Genus: Flavivirus
• Type virus: Dengue Virus
 Ref: Antiviral Res. 2009 January; 81(1): 6–15.

LIFE CYCLE OF DENV In the low pH of the Trans-Golgi Network (TGN),

prM is cleaved by furin

attachment

endocytosis

After replication, assembly

occurs in ER membrane.
Encapsidation (produces capsid and RNA)
result in immature virion
 Virology Journal 2005: 2; 26

ELECTRON MICROSCOPE
MORPHOLOGICAL
OBSERVATIONS OF DEN2
VIRUS PARTICLES
Antiviral Res. 2009 January; 81(1): 6–15.
DENGUE TIPE 1 DISTRIBUTION PLoS One. 2013; 8(5): e62649.
3 FACTORS DETERMINING SEVERENESS OF
DENGUE INFECTION
HOST FACTOR
• For example: During the epidemic of DHF/DSS in Cuba
infected Europeans were more severely ill than African
• Different alel class I of HLA shows different
susceptibility to DHF.
OTHER DISEASE AND THE ROLE OF AGE
• Diseases that are correlated with severeness
DHF/DSS: asthma bronchial, diabetes mellitus,
peptic ulcer, and sickle cell anemia
• Age:
• South East Asia > children than adults, probably due
to more permeable vascular endothelial among
children.
• Other countries found > more in children, some
countries found more in adults.
 DHF/DSS: AUTOIMMUNE RESPONSE
• Antibodi terhadap serotype-specific bersifat protektif seumur
hidup terhadap serotype yang sama (homolog), dan protektif
silang terhadap serotipe yang beda (heterolog) selama 34
bulan.
• Antibodi terhadap protein E pada permukaan virus bereaksi
silang dengan plasminogen dan menyebabkan perdarahan,
dan Ab anti-DENV NS1 bereaksi silang sehingga merusak
platelet (trombosit) dan sel endotel inang
• Respon imun tjd terhadap berbagai komponen DENV pada
DHF/DSS dengan bukti adanya immune activation markers (IL-
6, IL-8, TNFa, IFNg, dan komponen 3A dan 5A complement)
dengan perubahan fungsi platelet, DC, monosit, dan sel T.

Viral Immunol. 2006 Summer;19(2):127-32.

 NEW INSIGHTS INTO MECHANISME OF
THROMBOCYTOPENIA
• Bone marrrow suppression and destruction of thrombocytes
• Dengue virus is present in mature thrombocytes  clearance by
phagocytic cells
• Dengue-exposed thrombocytes exhibit markers of apoptosis
• Anti-NS1 antibody cross react with platelets  inhibits platelet’s function
• Dengue virus infects the endothelial cells  increased permeability
• Cells treated with DENV-EIII (protein envelope domain III) showed
suppressed megakaryopoiesis, alters autophagy and increases markers
associated with apoptosis
Ref: Goldthorpe SC and Conway MJ, 2017
ANTIBODY DEPENDENT ENHANCEMENT
• ADE terjadi pada masa protektif silang (infeksi kali ke2):
• Antibodi yang sudah ada thd satu tipe Dengue karena infeksi
sebelumnya namun dlm konsentrasi non/subnetralisasi terhadap
virus heterolog (virus serotipe beda yang menginfeksi sekarang)
akan melekat pada DENV (membentuk imunkompleks Ab-DENV)
dan akan meningkatkan terikatnya DENV dengan sel yang
mempunyai FcR , kemudian virus bereplikasi di dalam sel tersebut,
mengaktivasi komplemen sehingga dilepaskan banyak sitokin
dengan target sel endotel pembuluh darah.
• ADE terjadi pada beberapa jenis sel yang mempunyai FcR
yi human plasmacytoid DC, mature DC dan monosit.
• Selain ADE ada faktor lain (respons imun innate) yang ikut
berperan menentukan beratnya gejala klinis dan banyaknya
VL .
Virology. 2011 Dec 20;421(2):245-52.
DENGUE IN INFANTS

FEMS Immunology & Medical Microbiology Volume 59, Issue 2, pages 119–130, July 2010
 THE ROLE OF CYTOKINE, CHEMOKINE AND
LEUKOCYTE IN DENGUE PATHOGENESIS
Virus hati dan limpa  bereplikasi dan
menyebabkan inflamasi  banyak kemokin
CC di dalam darah, hati dan limpa 
mengaktivasi sel (melalui reseptor kemokin)
kerusakan jaringan hati/limpa.

Lekosit yang teraktivasi (terutama

limfositTh17/Th22dan trombosit) ini bersama
kemokin CC masuk ke hati.

Sel iNKT yang direkrut karena memiliki

reseptor CCR2 dan CCR4 menyebabkan
kerusakan hati.

Sel NK menghasilkan IL22 dalam hati. Sel T

gammadelta menghasilkan IL-17 bersama
kemokin CC menyebabkan proses inflamasi.

Immunology. 2013 Oct 12.

 LABORATORY TESTS FOR DEFINITIVE
DIAGNOSIS
Either one:
• Nucleic Acid Testing (PCR)
• NS1 (nonstructural protein 1) in serum
• VIRAL ISOLATION (tissue culture)
• IgG seroconversion or significant increase of antibody (by a
qualitative assay)
• 4 fold increase of antibody (quantitative assay) in paired sera
which is proven by neutralization test or another specific test
• Dengue specific IgM in Cerebrospinal Fluid (in the absence of IgM
to Murray Valley Encephalitis/Kunjin/Japanese encephalitis virus)
• Dengue specific IgM which is confirmed by a reference lab.
TIME SEQUENCE OF DENGUE INFECTION
 TIME OF LABORATORY TESTING
• PCR and NS1 : day -1 to 5
• IgM Ab (ELISA) : from d 3
• IgG Ab (ELISA) : from d 6

www.health.qld.gov.au
 AVAILABLE TESTS
Five serological tests:
1. hemagglutination-inhibition (HI)
2. complement fixation (CF)
3. neutralization test (NT)
4. immunoglobulin M (IgM) capture enzyme linked immunosorbent assay (MAC-ELISA)
5. Indirect immunoglobulin G ELISA.
The limitations of these techniques are the high cross-reactivity observed with these tests.

Four methods of viral isolation:

1. intracerebral inoculation of newborn mice
2. inoculation on mammalian cell cultures
3. intrathoracic inoculation of adult mosquitoes
4. inoculation on mosquito cell cultures.

Molecular diagnostic techniques:

1. nucleic acid hybridization
2. RT-PCR.

The Brazilian Journal of Infectious Diseases 2004;8(6):390-398

Incidence/Epidemiology
Virology/Taxonomy/Genomic
Virulence/Host Immune response
Laboratory Diagnosis
Prevention/Control

HUMAN IMMUNODEFICIENCY VIRUS

HIV
• Simian Immunodeficiency Virus (SIV) terdapat pada 26
spesies primata non-human di Afrika
• Dua dari SIV diketahui dapat menyebabkan HIV pada
manusia:
• HIV-1 – SIVcpz dari chimpanzee
• HIV-2 – SIVsm dari sooty mangabey
• Lentivirus ditransmisikan ke manusia 8 kali

Emerging Infectious Diseases 2002; 8: 451-7

EPIDEMIOLOGY

http://aids.gov/hiv-aids-basics/hiv-aids-101/global-statistics/
 HIV CRISIS: WORLD’S LARGEST CHALLENGE
• 33.4 millions of people died with HIV/AIDS
• >25 millions died due to AIDS since 1981
• 2008: 2 million died but 2.7 million new cases identified.
• 97% patients reside in low-moderate especially sub-Sahara
Africa: a country which cannot prevent the spread.
• No cure (ARV only supresses viral replication)
 HIV CURE
• Functional cure: When after
the patient stop ART their HIV
remains in remission and does
not damage their immune
system enough to cause any
adverse health consequences.
• Sterilizing cure: the complete
eradication of all HIV from a
Examples of HIV functl’ cure:
person’s body.
The Berlin Patient (2008)
The Mississippi baby (2013)
The VISCONTI Cohort (2014)
Visconti=virological and Immunological Study on Controllers after Treatment Interrruption
Silencing dgn
hipermetilasi DNA pada
gen promoter dan
enhancer virus

Liu, 2017
ELITE CONTROLLER OF HIV
• = Are a unique subset of HIV-infected individuals who
spontaneously control HIV  'functional cure' (long
term control of viral replication and remission from
symptoms of HIV infection in the absence of
antiretroviral therapy)
MAN FUNCTIONALLY CURED FROM HIV

• The Berlin Patient – Timothy Brown was an American who

was diagnosed as HIVpositive in Berlin in 1995, received ART
for 10 years and then was diagnosed with AML (acute myeloid
leukemia). He then was transplanted stem cells from a
Caucasian donor who genetically lack the CCR5 gene encoding
the host cell’s HIV co receptor) and had claimed HIV cured in
2008.
The ‘Berlin patient’ has shown that the re-engraftment of
bone marrow stem cells with certain genetic
traits can cure AIDS and at least inhibit HIV infection in the
long term to below detectable levels.
THE MISSISSIPPI BABY

• The baby girl born in 2010 was given ART at 31 days

after birth from a HIV pos mother, given till 18 months,
ART stopped due to lost from care, after 28 month
without ART she remained with no detectable HIV RNA
and was reported as functionally cured (March 2013).
• Unfortunately Rebound was detected in this baby at
the age of 4 years old (July 2014!!!)
THE HIV CURED BABY REBOUND
VISCONTI COHORT
• 14 persons in France (diagnosed HIV pos from
between 1996-2002) who were soon started ART
(during acute phase), given for at least 1 year and then
stopped, they showed undetectable virus RNA for 4
years now.
 NATION AND GLOBAL CONCERN

• HIV berdampak pada penderita; rumah tangga/keluarga;

masyarakat; dan perkembangan ekonomi bangsa
• Keberhasilan sampai saat ini:
• Adanya upaya global mengatasi HIV
• Prevalensi agak menurun di banyak negara  infeksi
baru berkurang
• Penderita yang mendapatkan ARV di negara miskin
meningkat 10 kali lipat (sampai 4 juta pada thn 2008)
 TAXONOMY/CLASSIFICATION OF HIV

• HIV-1 is most common world wide.

• “HIV” refers to HIV-1.

• HIV-2 reported only in Africa; clinical symptoms are the

same but HIV 2 progresses slower.
 SUBTYPES OF HIV
Group P
Alfa, beta,
gamma
HIV-1 Group O
ORTHO
RETRO Lentivirinae
Group N
VIRIDAE HIV-2
RETRO Group M Subtype A,B,C,D,F,G,H,J,K,CRFs
VIRIDAE Delta, epsilon SIV
SPUMA
RETRO SPUMAVIRUS
VIRIDAE

subtypes in Indonesia:
CRF_AE and B
 RNA Capsid Lipid layer
HIV STRUCTURE Enzyme Capsomer Glycoprotein
 HIV-1-INFECTED MT-4 CELLS
infected cells with budding structures and immature virus.

mature extracellular HIV-1 particles a higher-magnification view of an immature particle still

connected to the cellular membrane
IMPORTANT ENZYMES OF HIV
Enzymes for viral replication:
• Reverse transcriptase – tRNA serves as primer
• Integrase –
• Protease –
• Ribonuclease --
 GENE AND GENE PRODUCTS OF HIV
Protein Gene Product Function
Structural Gag P17 (matrix) Core Protein
P24 (capsid)
P7(nucleocapsid)
Env Gp120 envelope
Gp41
Catalytic Pol Protease Enzyme
Rev transcriptase
Integrase
Regulator Tat Tat increase mRNA production
Rev Rev Transport mRNA
additional vpu Vpu Degrade CD4, viral release
vif Vif Inhibit cell action that inhibits virus
vpr Vpr Import to nucleus
nef Nef HIV replication within Ly T and monocyte
 CELL RECEPTOR FOR HIV
HIV must attach to CD4 receptor and co-receptor:
Co receptor CCR5 is found on macrofage and T lymfocyte; CXCR4 is found on T lymphocyte.

Virus entry No Virus entry

enabled due is due to the
to availability chemokine co-
of CD4 receptor (CCR5
receptor and or CXCR4) is
a chemokine engaged in the
co-receptor binding to its
ligand

Ref: Sultan Qaboos Univ Med J. 2007 August; 7(2): 82–96.

HIV ENTRY INTO CELL – DRUGS TO INTERFERE HIV

HIV entry starts by Gp120 binding to CD4 receptor changes the

conformation and causes Gp120 binds to co-receptor CCR5 or
CXCR4  triggers membrane fusion and forms six-helix bundle.

From:Ref: Chukwuka et al, in Viruses. 2012 February; 4(2): 309–324.

 HIV REPLICATION
2. RNA, enzim RT, integrase,
1. Fusi virus pada protein lainnnya
permukaan sel Masuk ke dalam sel

3.Pembentukan DNA virus

4. Masuk ke nukleus dan

alami integrasi

7. Maturasi terjadi; virion 5. RNA baru dari virus

siap keluar sel berperan dalam
pembentukan protein

6. RNA virus baru beserta protein yang diperlukan

dibawa ke permukaan sel (bentuk imatur)
 DEVELOPING ARV DRUGS
Eg. fusion inhibitors =Enfuvirtide

Eg. co receptor antagonist =Maraviroc

ARV= antiretrovirus
HIV RESISTANCE TO ARV DRUGS
For further Reading: HIV Drug Resistance: New Insight and Updated Practices
Author: Daniel R. Kuritzkes, MD
 http://www.ncbi.nlm.nih.gov/books/NBK2245/

HIV GENETIC VARIATIONS

• Genetic variations among subtypes of HIV:
• Allow virus evade the immune system
• Persistence
• Antiviral resistance

In subtype B, only one nucleotide change is required to switch from the wild-type (WT) valine to the mutant
adenine, whereas in subtype G two nucleotide changes are required (a).

Adenine will therefore be the preferential drug-resistant substitution in subtype B.

In subtype G, resistance is preferentially acquired by changing the WT isoleucine for the mutants threonine
or methionine, requiring a single nucleotide change (b).
QUASISPESIES
• Quasispesies: variasi pada virus (mutan) yang berbeda-beda
pada satu atau lebih gen
• Akibatnya populasi virus yang menginfeksi tubuh inang terdiri
atas virus yang tidak identik
• Memungkinkan virus tertentu lebih mampu beradaptasi
• Sekuens Konsensus: sekuens yang umum
• Sekuens Master: sekuens paling fit (kuat) dalam suatu
lingkungan/environment
Haase AT, Nature 464, 217-223(11 March 2010)
HIV TRANSMISSION VIA MUCOUS MEMBRANE
Viruses enter
through minor
lesions on the
mucous layer;
bind to CCR5 (on macrophage
and Lymphocytes)
and CXCR4 (on Lymphocytes)
 Or binds or enter DC
 Enters Lymphnode (many
Tcells but less viral entry
inhibitors)

 SEXUAL TRANSMISSION
MUST BE BLOCKED
FROM CONTACT WITH
submucosal lymphocytes’
CD4/ co receptors and DC

Ref: Davis and Doms, J Exp Med. 2004 April

19; 199(8): 1037–1040
PREVENTION OF HIV TRANSMISSION
•

Ref: Lancet. 2013 Nov 2;382(9903):1515-24.

HIV RESERVOIR IN THE BODY  THE MAIN
OBSTACLES TO VIRAL ERADICATION

• HIV-1 remains latent in resting CD4+ T Lymphocytes (In Vivo)

• Blood-tissue barrier inhibits ARV drugs entering the CNS, the
retina, and the testes.
• Cryptic viral replication in infected CD4 lymphocytes
• Virions bound to dendritic cells in the lymphnode persist for a
long time
Incidence/Epidemiology
Clinical Appearance
Virology/Taxonomy/Genomic
Virulence/Host Immune response
Laboratory Diagnosis
Prevention/Control

POLIOMYELITIS VIRUS
 POLIOMYELITIS AKUT
• WHO 1988: eradikasi global poliomielitis paralitik sblm 2000
• Strategi:
• 1) vaksinasi massal dgn vaksin live attenuated oral pd anak <5 th
• 2) surveillans
• 3) imunisasi pada daerah/populasi yang kemungkinan masih terdapat penyebaran
polio
• Kenyataan: Virus wild-type polio masih terus ada pada daerah endemik, dan kadang2
pada daerah yg dianggap sudah bebas polio.
• Sindrom deteriorasi fungsional terlambat dapat berlangsung selama bbrp tahun setelah
poliomyelitis paralitik akut atau 'post-polio syndrome’ gejala motor neuron terjadi
setelah bertahun2 infeksi dimasa kecil – salah satu contoh: Franklin D. Roosevelt alami
infeksi pada usia 39 tahun dan munculnya gejala lambat lainnya setelah itu
 DETERIORASI
FUNGSIONAL
Akibat kelemahan otot
dan
gangguan bentuk rangka tubuh
PERJALANAN PENYAKIT
• Reservoir : GIT
• Rute infeksi: oral - oral dan faecal - oral.
• Viruses multiplikasi dalam farings dan usus masa inkubasi
1-3 minggu – diseminasi darah – seluruh tubuh
• Eksresi saliva 2-3 hari, pada faeces 2-3 minggu kemudian.
• Sifat infeksius ada 7-10 sebelum dan setelah gejala muncul.
• 95% infeksi virus ini asimptomatik atau seperti flu-like'
illnesses yangsembuh sendiri
POLIOMIELITIS NON-PARALITIK ATAU
PREPARALITIK
• Setelah gejala prodromal, pasien mengalami demam
tinggi, faringitis, myalgia, anorexia, nausea dan muntah-
muntah, dan sakit kepala, kaku leher (meningitis).
• Gejala ini menghilang dalam 1-2 minggu.
POLIOMIELITIS PARALITIK
• Setelah fase meningitis  poliomyelitis tipe spinal (nyeri otot hebat,
kadang dengan kejang otot, melemah asimetris, sensorik baik hanya
paraestesi, sering pada tungkai saja dan fasikulasi).
• Bentuk bifasik: ketika kelemahan otot baru terjadi setelah melewati
periode singkat dimana kondisi pasien membaik.
• Infeksi tanpa kelemahan otot bisa terjadi terutama pada anak-anak yang
sudah diangkat tonsil/adenoidnya
• Orang dewasa bisa bergejala spinal. Infeksi ke medulla bisa
menyebabkan disfagi, disfonia dan gagal nafas
• Gangguan Vasomotor (hipertensi, hipotensi dan kolaps sirkulasi
menyebabkan kematian).
• Gangguan kencing dan gangguan BAB.
• Tipe encefalitik: agitasi, confusion, stupor, coma.
RANCHO LOS AMIGOS REHAB CENTRE 1953

J Neurosci. 2013 January 16; 33(3): 855–862.

TAXONOMY/CLASSIFICATION
• Family: PICORNAVIRIDAE
• Genus: Enterovirus
• Type name: Poliovirus
FOTO TEM POLIO VIRUS
. J Neurosci. 2013 January 16; 33(3): 855–862

• 30 nm
 STRUCTURE OF
POLIOVIRUS
• Pos strand ss RNA

J Neurosci. 2013 January 16; 33(3): 855–862.

 Proc. Jpn. Acad., Ser. B 83 (2007)

PATHOGENESIS INFEKSI POLIOVIRUS

 Proc. Jpn. Acad., Ser. B 83 (2007)

TRANSPORT RETROGRAD PADA AXON

STRATEGI VIRUS BERTAHAN DI DALAM SEL
NEURON
• Sel neuron dapat membatasi replikasi dan penyebaran virus ini  virus
hilang tanpa banyak neuron rusak atau terjadi infeksi persisten non
sitolitik.
• Dugaan mekanisme terjadinya post polio syndrome
• Reaksi IgM thd virus yang baru muncul lagi belakangan hari (terbukti
dengan pemeriksaan PCR positif)
CYTOPATHIC EFFECT (CPE) PADA SEL NEURON

Proc. Jpn. Acad., Ser. B 83 (2007)

DETEKSI
• Sel kultur dengan rhabdomyosarcoma cell line (RD cells) dan mouse L cell line yang
mempunyai PV receptor (L20B cells)
• Keuntungan:
• 1) tidak perlu alat molekuler
• 2) sangat sensitif (detection limit 1 infectious dose yg mengandung 50 - 1,000
virion
• Kelemahan:
• Perlu keahlian dan kontrol kualitas sistem kultur dalam mengevaluasi CPE.
• time-consuming: 10 hari
DETEKSI BMC Infect Dis. 2009 Dec 16;9:208

• Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) system.

• Keuntungan:
• 1) Peralatan minimal -- isothermal heat bath
• 2) sangat sensitif (detection limit 0.01 PFU utk respiratory syndrome
coronavirus, 0.1 PFU utk mumps virus, 0.4 focus forming units utk hepatitis A
virus, 50 copies utk swine vesicular disease virus)
• 3) cepat (1 jam)
• 4) kurang kemungkinan kontaminasi silang antar sampel.
DETEKSI J Clin Microbiol. 2010 August; 48(8): 2698–2702.

• Particle agglutination

(A) Schematic view of a sensitized gelatin particle with a

soluble PVR (PVR-IgG2a).
(B) The order of sample addition to the reaction plate is as
follows: 1, anti-PV antibodies; 2, PV solution; and 3,
sensitized gelatin particle solution. r.t., room temperature.
 Attenuated Polio Vaccine

Vaksin Polio : Polio 1-2-3

Sept 2015 Poilio 2 declared eradicated. Polio 3 never detected since 2012
 PENCEGAHAN
• Salk trivalent inactivated polio vaccine: tahun 1956: PER
INJEKSI  dapat mengstimulasi IgM, IgG and IgA tidak
mengstimulasi secretory IgA.
• Sabin trivalent oral live attenuated polio vaccine:
1962mengandung poliovirus I, II and III yang tumbuh dalam
sel kultur. PER ORAL  infeksi aktif orofarings dan endotel
usus shg IgA dihasilkan juga.
• Timbulkan herd immunity karena disekresikan lewat feses.
• Dewasa yg lahir sblm 1958 dan belum imunisasi harus
diimunisasi -- Tiga dosis: 2, 3, 4 bulan
• Eradikasi global tercapai bila semua orang berhasil divaksinasi
GLOBAL ERADICATION OF POLIOVIRUS
• Withdraw trivalent OPV (as of August 2016)
• Replacement with b OPV (contains polio 1 and 3 only)
• Include 1 trivalent inactivated in 1 of the schedule
• 2016- 89% of world countries have implemented
 ALFAVIRUS
Tipe Nama lain Sel target Tempat laten patofi Transmisi

Human HSV-1 Oral dan atau

Herpesvirus 1 genital (utama
oro-fasial) Kontak
Human HSV-2 Oral atau langsung (oral
Herpesvirus 2 genital (utama atau seksual)
Mukoepitel Sel saraf genital)
Human VZV Cacar air(Ch Respirasi atau
herpesvirus 3 pox) atau kontak
Cacar api langsung
(Shingles)
Incidence/Epidemiology
Clinical Appearance
Virology/Taxonomy/Genomic
Virulence/Host Immune response
Laboratory Diagnosis
Prevention/Control

HUMAN HERPES VIRUS 1 & 2

 HSV INFECTIONS
Foto: FatahzadehM, 2007

Primary herpetic ginggivostomatitis

in a young child

Herpes labialis Recurrent herpeslabialis in

Immunocmprs patient

Recurrent intraoral herpes

Primary herpetic whitlow in

Genital herpes in
2 yrs old with primary oral
Immunocompetent person
erpes
Recurrent intraoral herpes in immncmprs patient
TAXONOMY/CLASSIFICATION
• Ordo: Herpesvirales
• Famili: Herpesviridae
• Subfamili:
• Genus:
Alfaherpesvirus
Virus Simpleks
 HSV STRUCTURE
• Icosahedral
• Out to inside:
• Envelope + glycoprotein knobs
• Tegumen
• CAPSID
• DNA
GENOM HSV: 152000 BP

Daerah unik panjang dan pendek, diapit dengan sekuens berulang

VIRUS REPLICATION OF HSV-1
GARPU REPLIKASI VIRUS HSV-1
A. Molekul DNA untai ganda terlepas oleh H/P.
Polymerase HSV (oval hitam didalamnya ada UL30
oval dan UL42 bentuk bulan sabit) menyebabkan
sintesis DNA leading strand. Dibawah H/P ada RNA
primer .
ssDNA keluar dari balik helicase terbungkus ssDNA-
binding protein ICP8.

Terbentuk replication loop pada lagging strand agar

dapat bersesuaian dengan leading strand.
DNA polimrease pada Strand lagging memulai
okazaki fragment dengan menggunakan primer RNA.
 ACTIVATION OF ALFAVIRUSES

Neurotropic and neuroinvasive virus

PRODUCTIVE VS
LATENT
• Jika ada IE ICPO protein, enzim repair DNA
terhambat, genom tetap linier = virus produktif.
• Jika ada enzim repair DNA maka genom
menjadi sirkuler= laten.
 JALUR INFEKSI ALFAHERPESVIRUS PADA
SISTEM SARAF MAMALIA

Alfaherpesvirus:
•HSV-1
•HSV-2
•VZV

• Infeksi biasa berawal pada perifer yakni epitel mukosa  virus masuk pada bagian terminal neuron
sensorik pada sistem saraf perifer (PNS) ; secara retrograd sepanjang akson mencapai badan sel, dan
disimpan (seumur hidup) dalam inti sel.
• Ketika ada reaktivasi, partikel virus baru dibentuk dan terletak dekat lokasi akan dikeluarkan dari sel.
Infeksi menyebar ke arah anterograd ke arah perifer.
• Infeksi juga bisa berlangsung secara trans-neuronal, dari PNS ke CNS  fatal encephalitis .
SEQUENTIAL OF HERPES SIMPLEX VIRUS
INFECTION

Legoff et al, 2014, Virology Journal, 11:83

GENOM HERPES SIMPLEKS 2 (HSV-2)

Gambar ds-DNA HSV-2

Daerah Unik: UL dan US
Bagian Repeat Mayor: TRL, IRL, IRS

Ref: Dolan etal. 1998

 Ref: Anderson, 2012

HSV 1 AND HSV 2

• Acute – chronic disease
• Mild – severe disease
• No curative therapy
• Lifelong carriage
• Recurrent outbreaks
DIAGNOSIS OF HSV
• Cell culture – microscopy
• ELISA – only for past exposure (not acute)
• PCR
 DIRECT LABORATORY METHODS FOR HSV
DIAGNOSIS
Method Principle Sample
VIRAL ANTIGEN Immunoperoxidase staining Swab, smears from lesion, smear or
DETECTION vesicular fluid or exudate fr. base of vesicle
Swab, or vesicular fluid or exudate fr. base
Capture ELISA of vesicle
Swab, or vesicular fluid or exudate fr. base
Rapid Test Device of vesicle

VIRUS CULTURE HSV Isolation susceptibel test Swab, Skin Lesions, vesicular fluid or
exudate fr. base of vesicle, Mucosal sample
w.o. Lesions biopsies, conj./corneal Smears,
neonates
MOLECULAR HSV DNA Detection and or quatitation Swab, Skin Lesions, vesicular fluid or
BIOLOGY by NAAT/inhouse classical exudate fr. base of vesicle, Mucosal sample
PCR/Realtime PCR/ commercial PCRs w.o. Lesions biopsies,
Aqueous/Vitreous humor, CSF, blood
CYTOLOGICAL Tzank smears Skin/mucosal lesions.
EXAMINATION Papanicolau or Romanovski stain Biopsies, Conjunctival/mucosal smears.
Direct Immunofluorescence Smears, tissue section smear fr base of
vesicle.
INDIRECT LABORATORY DETECTION OF HSV
(SEROLOGICAL TESTS)
Method Principle Sample
Western Blot Western Blot HSV 1 Serum
Western Blot HSV2
EIA (enzyme immunoassay) Monoclonal Antibody Serum
Blocking EIA
ELISA
POC (Point of Care tests) Immunofiltration Serum, capillary blood
HERPETIC WHITLOW
Incidence/Epidemiology
Clinical Appearance
Virology/Taxonomy/Genomic
Virulence/Host Immune response
Laboratory Diagnosis
Prevention/Control

HUMAN HERPES VIRUS 3 (VARICELLA ZOSTER

VIRUS)
GENOM HHV3: 124884 BP
 STRUCTURE OF VIRUS VARICELLA ZOSTER
• Virion size 80-120 nm
(Rash without pain)
VZV (VIRUS VARICELLA ZOSTER)
 HERPES ZOOSTER = SHINGLES

Zoster after given Mycophenolate Mofetil (antimetabolit) in Sarcoidosis patient

Ref: Hegde,2012
HERPES ZOSTER = SHINGLES

Ref: Sampath Kumar, 2009

 HERPES ZOSTER DUPLEX BILATERALIS :
RARELY OCCURRING
• Case report (in) A 15-year-old Chinese boy presented with a bilateral
and symmetrical painful eruption on the upper abdomen of 7 days'
duration. The eruption was preceded by a 2-day history of malaise and
low grade fever. He did not have the varicella vaccine but had
chickenpox at 3 years of age. His past health was otherwise
unremarkable. In particular, he did not have recurrent or chronic
infections. The patient did not have recent weight loss and was not on
any medications. There was no history of recent travel. He did not
have exposure to venereal or other infectious diseases. The family
history was noncontributory. (Ref: Leung et al, 2015)

Previously the same case was reported in 1947 (Ref: Thomas), in 2003
(Ref: Arfan ul-bari ), and in 2006 (Ref: Brandon)
Incidence/Epidemiology
Clinical Appearance
Virology/Taxonomy/Genomic
Virulence/Host Immune response
Laboratory Diagnosis
Prevention/Control

MUMPS VIRUS
FAMILY PARAMYXOVIRIDAE
• Genus Morbillivirus  morbillivirus
• Genus Rubulavirus  mumps virus
• Genus Pneumovirus  RSV
INCIDENCE / EPIDEMIOLOGY
• 1st described by Hippocrates (5th Century)  then in 1930, Johnson and
Goodpasture inoculated parotid tissue from Macaca mulatta into some
children.
• World wide distribution
• Causes encephalitis, meningitis, orchitis, myocarditis, pancreatitis and
nephritis
• Global resurgence of cases were reported in highly vaccinated populations
• Neurotropic  50% cases are CNS associated
• Cases of aseptic meningitis are associated with some vaccine strains
MUMPS ETIOLOGY
• Family: Paramyxoviridae
• Genus: Rubulavirus
• Type name: mumps virus
 MUMPS VIRUS STRUCTURE
• Negative sense, single stranded (non segmented) RNA virus
• 15.384 nucleotides long
• Helical/pleomorphic particle 120-450 nm (average 200 nm)
• Enveloped
• Serotypes: A-N (A, B, C, D, F, G, H, I, J, K, L, N) excluding (E and
M).
• Vaccine strains: A, B, or N
 MUMPS CLINICAL SYMPTOMS
 Fever
• Headache
• Muscle aches
• Tiredness
• Loss of appetite
• Swollen and tender salivary
glands under the ears or jaw
on one or both sides of the
face (parotitis)
• Long-term sequelae: paralysis,
seizures, cranial nerve palsies,
hydrocephalus and deafness

Long-term sequelae: paralysis, seizures, cranial nerve palsies, hydrocephalus and deafness
Rubin S, et al, J Pathol 2015; 235: 242–252
www.CDC.gov
E.M. OF MUMPS VIRUS

• EM of mumps virus by dr. F.A.Murphy (1976, CDC)

Hemagglutinin and Neuraminidase protrude from the envelope

MUMPS VIRUS PATHOGENESIS
AND THE UNRESOLVED QUESTIONS

Rubin S, et al, J Pathol 2015; 235: 242–252

 LABORATORY METHODOLOGY FOR MUMPS
DIAGNOSIS

1. Virus Isolation (Gold standard)

2. Nucleic acid detection (New gold standard)
3. Serological confirmation (IgM detection)
METHODOLOGY
• Inoculation of specimen into cell cultures  Incubation of
cultures  observation for characteristic cytopathic effect, and
• Followed by identification by DFA or other methods

Photography by Linda Stannard, in www.virologyonline

Incidence/Epidemiology
Clinical Appearance
Virology/Taxonomy/Genomic
Virulence/Host Immune response
Laboratory Diagnosis
Prevention/Control

MORBILLI VIRUS
VIRUS CLASSIFICATION
• Family: Paramyxoviridae
• Genus: Paramyxovirus
• Type name: Morbillivirus
DISTRIBUTION OF MEASLES GENOTYPES
MEASLES VIRUS STRUCTURE
 Griffin et al 2012

MORBILLIVIRUS LIFE CYCLE

MORBILLI RASH
• Rash is due to leukocytes, CD4 and CD8 T Lymphocytes infiltration

Griffin, et al, 2012

LABORATORY METHODOLOGY FOR MEASLES
VIRUS DIAGNOSIS
1. Virus culture : observe CPE
2. Nucleic acid detection
3. Serological confirmation (IgM detection)

Photo by Linda Stannard, University of Cape Town, S.A in www.virology-online.com

Incidence/Epidemiology
Clinical Appearance
Virology/Taxonomy/Genomic
Virulence/Host Immune response
Laboratory Diagnosis
Prevention/Control

RESPIRATORY SYNCYTIAL VIRUS

VIRUS CLASSIFICATION
• Family: Paramyxoviridae
• Sub family: Pneumovirinae
• Genus: Pneumovirus
• Type name: Respiratory Syncytial Virus
VIRAL CHARACTERISTIC OF RSV
• Enveloped
• Spherical or long filaments
• Negative sense single stranded RNA
• 15.222 nucleotides
• 10 genes encoding 11 proteins
STRUCTURE AND GENOME ORGANIZATION OF
RSV – SPHERICAL AND FILAMENT SHAPED

10 genes of RSV encodes 11 proteins

Lambert L, et al, 2014 Front Immunol. 2014; 5: 466.

INCIDENCE AND EPIDEMIOLOGY
• A novel virus was isolated from chimpanzee with respiratory
symptoms in 1955, was named chimpanzee coryza agent.
• Soon was understood as human virus, renamed as RSV
(because it forms the giant syncytia in tissue culture)
• RSV is the most common cause of severe respiratory infection
in infants and young children.
• RSV is also important pathogens among:
• Elderly
• Those with underlying lung disease
• Those with impaired immunity
INFECTION AND REINFECTION OF RSV
• Highest incidence at 2-3 months old babies.
• World wide 33.000.000 RSV cases; 3.400.000 death; Mortality
rate 66.000-199.000/year (Lanari et al, 2005).
• Able to reinfect repeatedly although no change of viral
antigenicity, but previous infection will cause reduced severity.
• Such reinfection is due to the ability of RSV to inhibit or
subvert host immune system
RSV LIFE CYCLE
RSV spread from
cell to cell by
causing cell fusion
and
syncytia formation

Bawage et al. 2013, Advances in Virology vol 2013

RSV INFECTION TRIGGERS CELL RESPONSE

Openshaw and Tregoning, CLINICAL MICROBIOLOGY REVIEWS, July 2005, Vol 18, No 3, p. 541–555
LAB METHODS FOR DIAGNOSIS
1. Tissue Culture / virus isolation
2. Direct Immunofluorescence or IFA or EIA
3. Chromatographic Rapid Antigen Detection by Rev Transf PCR
4. Antigenic Capture ELISA, serology ELISA (IgM and IgG), HA,
HAI, Neutralization test.
 INFECTED CELLS FUSE AND FORM SYNCYTIA
day 5 fused cells are dead and
Day 1 no change in cells detaches from the base of plate

day 3 some cells fused and form syncytia

Hep2 cells = laryngeal carcinoma cell line

Domachowske et al, Clini Microbiol Reviews Vol 12 no 2, 1999

CELL CULTURE FOR DIAGNOSIS OF RSV

Uninfected Hep-2 cells RSV Infected Hep-2 cells

Leland and Ginocchio, Clinical Microb Rev, Vol 20 no 1, 2007, p 49-78

Incidence/Epidemiology
Clinical Appearance
Virology/Taxonomy/Genomic
Virulence/Host Immune response
Laboratory Diagnosis
Prevention/Control

MOLLUSCUM CONTAGIUM VIRUS

MOLUSCUM CONTAGIOSUM
• Family Poxviridae genus Molluscipoxvirus.
• double-stranded DNA genome of 190 289 bp
• Preadolescent children  sanitation, crowds, swimming pool
• Adolescent and adults  sexual transmission
• Self limiting disease – unless HIV/immunocompromise
 Hyperplastic, acanthotic squamous epithelium forming a central crater
filled with keratin fragments and molluscum bodies

Molluscum contagiosum
virion with envelope
measuring 340×265 nm

Inclusion bodies in
cytoplasma of cells
 PAPULES OF MOLLUSCUM CONTAGIUM
• Single -- multiple, 2-5mm sometimes , rounded, dome-shaped, pink, waxy, umbilicated with
caseous plug

Axilla of HIV infected man

Chest
Leg of HIV infected child

Cheek
MOLLUSCUM CONTAGIOSUM VIRUS

Ref: Chen 2013

Incidence/Epidemiology
Clinical Appearance
Virology/Taxonomy/Genomic
Virulence/Host Immune response
Laboratory Diagnosis
Prevention/Control

HUMAN PAPILLOMA VIRUS

VERRUCA VULGARIS (COMMON WARTS)
• Benign lesion of skin and mucous membranes caused by
human papillomavirus (HPV).
• The lesions are typically self-limited
• Vary in size, number, anatomical site, viral type.
• The benign VV: squamous papilloma with verruca vulgaris,
focal epithelial hyperplasia and condyloma .
• Most etiol.: HPV-2, HPV-4 or HPV-40.
VIROLOGY
• Small
• Non-enveloped
• Icosahedral viruses
• 50–60 nm in diameter
• Circular, doublestranded DNA genome (~7000–8000 bp).
Bosn J Basic Med Sci. 2014 Aug; 14(3): 136–138
 Oral cancer HPV +, shows highly
variable clinical features.

The most frequent appearance is

a white lesion or red or ulcerative
area.

Oral Implantol (Rome). 2015 Apr-Sep; 8(2-3): 45–51.

ASSOCIATION OF HPV AND CHILDREN
HPV VIRUS DISTRIBUTION ON SKIN LAYERS
HPV IN PAP SMEAR Clin Microbiol Rev. 1998 Apr; 11(2): 341–365.

HPV is identified predominantly in Pap smears; the hallmark of HPV infection is the
detection of koilocytes. The large, irregular, hollow cavity in conjunction with nuclear
enlargement and hyperchromasia and frequent binucleation are cytologic features of HPV
effect. PAP stain; magnification, ×1,000.
VACCINE
• HPV vaccine is the first explicitly designed to prevent virus
induced cervix cancer
• Women age 9-26 yo