Navid Dinparast Djadid PhD, Molecular Entomology, LSTM

Professor, Malaria & Vector Research Group (MVRG) Biotechnology Research Center (BRC),

Pasteur Institute of Iran (PII)

navidmvrg@gmail.com, navid@pasteur.ac.ir

On behalf of all MVRG members, especially Prof. S. Zakeri, Head of MVRG

MALARIA & VECTOR RESEARCH GROUP

MVRG

Iran Land Boundaries • total: 5,894 km • border countries: Afghanistan 921 km, Armenia 44 km, Azerbaijan 689 km, Iraq 1,599 km, Pakistan 959 km, Turkey 534 km, Turkmenistan 1,148 km Iran Coast Line: 2,440 km; and the Caspian Sea (740 km)

Elevation extremes

• lowest point: Caspian Sea -28 m highest point: Kuh-e Damavand 5,671 m

Doosti et al

In her PhD project identified two Aedes species in Baluchistan by morphological and molecular tools:

• Aedes albopictus

• Aedes unilineatus

• No Dengue virus was detected

Suggestions: entomologists should attend the field of VBDs

IPI, TUMS and other MSUs could work jointly, based on their expertise to urgently define:

• To map the distribution of different Aedes spp in the country

• To detect any pathogen (priority with arboviruses) infection within the Aedes spp

• To verify the phenotypic and molecular mechanisms underline the insecticide resistance/susceptibility

• To study the vector-pathogen interaction and the possible paratransgenesis control strategy

•

Dinparast Djadid N: Professor, Molecular Entomology Zakeri S: Professor, Immuno-Molecular Parasitology

Abouei A: Assistant Prof., Molecular Genetics Raz A: Assistant Prof, Medical Biotechnology

Tahghighi, A: Assistant Prof, Medicinal Chemistry Maleki-Ravasan, N: Assistant Prof, Medical Entom

Current Students: 8 Ph.D. & 11 M.Sc. Valizadeh V: Assistant Prof, Pharmaceutical Biotechnology

*Head of MVRG:

MVRG has 3 Certificates of Standards: ISO 15189, ISO 18001, ISO 13485

Malaria and Vector Research Group (MVRG)

Candidate WHO Collaborating Center (WHOCC)

&

National Reference lab for Molecular Diagnostics of Malaria

zakerimvrg@gmail.com

MVRG

• MVRG is an R&D Group:

Malaria and Vector Research Group (MVRG), Biotechnology Research Center (BRC), Pasteur

Institute of Iran (PII)

• Sites: 1- MVRG Research unit, PII Research Compos, Tehran

2- National Insectarium (MVRG R&D unit), PII production Complex, 20th km of Tehran-Karaj Highway

Malaria and Vector Research Group

(MVRG)

• Candidate WHO Collaborating Center

(WHOCC)

• National Reference lab for Molecular

Diagnostics of Malaria

• MVRG has 3 Certificates of Standards:

ISO 15189

ISO 18001

ISO 13485

MVRG Applied & Operational

Research & Activities:

1) Vaccine Development (Sexual and asexual stages against P. f. & P. v.)

2) Drug Resistance & Drug Discovery

3) Insecticide Resistance & Discovery

4) Paratransgenics and Transgenics in Anopheles-Plasmodium model

5) Therapeutic & Diagnostic Kit Production:

- Maggot therapy - Collagenase

- Molecular kit for Detection of Insecticide Resistance

- NASBA-ELISA kit for Plasmodium detection

6) Training 75 PhD & M.Sc. (Iran, Sudan, Iraq, Pakistan, Afghanistan)

& Public Health Staff from MSUs

7) Collaboration with 23 National and International Institutes & Universities

8) International Papers: 78 9) National & International Patents: 8

MVRG policies as National Reference Laboratory of Molecular Malaria (under supervision of Prof. S. Zakeri)

Assist the CDC in MOH and WHO/EMRO in malaria control and elimination campaigns:

Basic, applied and operational research:

• Molecular diagnosis of infected malaria parasite

• Immunology and molecular parasitology from lab to field

• Analyzing human-P. falciparum and P. vivax immune interactions

• Surveillance of anti-malarial drug resistance

Evaluation of malaria elimination programms:

• Providing baseline data for monitoring the impact of malaria control and elimination strategies using serology method

Training/workshop at the national and regional level

Set up standard guidelines and protocols for elimination strategies in Iran

Biotechnology-Related Laboratory Work on Malaria

After Field sampling • Microscopic examination of malaria parasites and vectors • DNA, RNA extraction • Designing different primers and probes • Different PCR methods for DNA, RNA amplification • Detection of coding and non-coding regions in genes • Cloning • Expression • Protein purification • McAb and Polyclonal antibody production • In vitro analysis • in vivo analysis in animal model • Bioinformatics analysis

Cloning of target gene in T-vector, Sub-Cloning in pQE30 T T A A

Recombinant T-vector

Transform of recombinant plasmid E. coli M15 and Expression

Demonstration of expressed protein by SDS-PAGE and western blotting

Purification and confirmation of expressed protein

Desalting of purified protein

Mice immunization

IFAT

ELISA Depletion assay Avidity ELISA

In vitro protein

expression

Erythrocyte inhibition assay

Blood and Sera samples

DNA Extraction from blood

PCR or Nested PCR

Electrophoresis

Sequencing, analysis

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Biotechnology & Vector Section

• Molecular systematics and population genetics of malaria vectors • Molecular mechanism/s of insecticide resistance in diseases vectors • Functional genome analysis in insect vectors of human diseases • Detection of Wolbachia in Culicidae mosquitoes: detection and possible use as

Gene driving system • Paratransgenics for mosquito control: detection and application of bacterial and

viral fauna in Culicidae mosquito and their genetic manipulation • Preparation on developing transgenic mosquitoes • Vector-Parasite interaction in malaria model • Sexual stage transmission blocking vaccines (TBV) • Developing molecular kit for detection of insecticide resistance in vector species • Developing molecular key to mosquito species (i.e. Iranian Anopheles species) • Larvae therapy

• Genetic composition of vector species: working on this subject provides the essential data on geographical distribution of main vectors, their subpopulations and vectorial status. Our studies so far led to development of a molecular key to Iranian Anopheles species, and detection of new members within main malaria vectors in Iran and Afghanistan.

• The composition of different pathogens (i.e. Plasmodium spp, …) within different vector species: this data will verify the role of each species in disease transmission. Our results through collaboration with neighboring countries revealed the presence of P. falciparum and P. vivax in Anopheles species from Iran, Afghanistan, and Iraq. Also, during the last 15 years we detected other agents such as viruses, bacteria and Wolbachia in Iranian Culicidae species, providing us the possibility to use them in planning the paratransgenic strategy for malaria control.

• Molecular mechanisms of insecticide resistance: this area of our research is a breakthrough in vector control strategy, as we managed to develop a molecular kit for detection of resistance/susceptibility to different generations of insecticides within vector species. Now, we are improving the specificity and sensitivity of developed kit by using new and field-friendly techniques.

• Developing vaccines that interrupt malaria transmission TBV/VIMT): Having access to malaria endemic areas and the construction of National Insectarium in MVRG, we were able to challenge the role of anti-WARP, Lectin, and CPB in blocking the P. falciparum and P. vivax sexual stage within An. stephensi in field and insectary, while the out coming results were promising and published in Malaria Journal and Infection & Immunity journals.

• Paratrangenics in malaria control: This strategy is one of those "edge of knowledge" research areas in MVRG, as when we started it in about 2007. we had no idea on the composition of different agents within the mosquito that might be used as paratransgenic agent. Now, by publishing partial results of our extensive field and laboratory studies in Plos One (2011), we are planning to deploy the target bacteria species in Anopheles-Plasmodium model. Detection of two types of Wolbachia and Denso viruse in Iranian Culicidae species also pave the way to use these agents in paratransgenics strategy and/or designing new gene driving systems.

National Insectarium (R&D and Training Centre for VBDs)

• Following WHO/EMRO request for MVRG assistance in

2005, this 450 square meters innovative infrastructure has been established

• It is fully functional since 2010 in PII, aiming to provide the edge of knowledge core facilities and expertise for working on different arthropod and insects transmitting pathogens that causing vector-borne diseases (VBDs) in order to:

a) Providing equal opportunity for all scientists in Iran, WHO/EMRO and PIs Network

b) Facilitate the cooperation and collaboration on genomics and applied studies for control of VBDs.

National Insectarium Pasteur Institute of Iran

2006

2010

The sections and laboratories in National Insectarium

• Insect Rearing Laboratories: Routine rearing of Culicidae mosquitoes (Anopheles, Culex, Aedes spp), and temporary project-based rearing of sand flies, fleas, ticks, myiasis flies, scorpions

• Morphological Studies Laboratory: identification, dissection, preparation of morphological keys, and comparative systematic

• Insecticides Laboratory: WHO routine susceptibility tests for different medically important arthropods

• Molecular Studies Laboratory: DNA, RNA extraction, PCR, RT, RFLP, cloning, expression, protein purification, bioinformatics, etc

• Cell Culture Laboratory: in vitro continues culture of P. falciparum, insect cell culture, …

• Microbiology Laboratory: Detection and identification of microbial flora within medically important arthropods as paratransgenic tools

• Challenge Laboratory for vaccine development • A “BioBank” for medically important arthropods and the pathogens they transmit • Lecture Room

Biobank

Anopheles and Culex rearing rooms

Morphology lab

Morphological identification of the main vectors

Culture lab

Drug Design and Proteomics lab

C T

C

Y

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Vector and parasite challenge labs

(transgenic & paratransgenic)

It is our responsibility: working together for their better future

AFRICA CHABAHAR, IRAN

Collaborators

Departments in PII CDC, Iran

MOH, IR Iran, Afghanistan, Pakistan, Sudan Medical Sciences Universities: Zahedan, Hormozgan, Kerman,

Guilan, Shiraz, Kashan, …

WHO/TDR WHO/EMRO

Karolinska Institute, Sweden Osaka Univ., Japan

Camerino University, Italy France: Institute Pasteurs’ International Network (RIIP),

Malaria Network, Pasteur Institute Paris, National Museum

Thanks for your

attention

Pasteur Institute of Iran