Arif Jamal Siddiqui, PhD Current Status: Post Doc Research Associate (TTUHSC Lubbock) E-mail: arif.j.siddiqui@ttuhsc.eduMobile Number: +1-8063178440Work Address: Department- Internal medicine, Texas Tech University Health Science Center, 3601 4th Street, Lubbock, Tx-79430.

CURRENT WORK: Currently I am working a most promising Sm-P80 vaccine against Schistosomiasis diseases in baboon’s model. Our aim and propose to make best vaccines against Schistosomiasis diseases with best research. EDUCATION: Ph.D., Thesis Submitted on June 2015 (Biological Sciences) to Academy of Scientific & Innovative Research, New Delhi, India (Research performed at CSIR-Central Drug Research Institute, Lucknow, India).Thesis title: “Studies on immunological responses elicited during Pre-Erythrocytic stage infection with rodent malaria parasite Plasmodium yoelii” Master of Science, 2005-2007. (Biotechnology), Punjab Technical University, Jhalandhar, India.Bachelor of Science, 2002-2005: (Botany, Chemistry, Zoology), VBS Purvanchal University, Jaunpur, India.PERSONAL DETAILS: Date of Birth: 1st May 1984, Nationality: Indian, Marital Status: Single

MAJOR SCIENTIFIC FIELD OF INTEREST: Immunology, Molecular cell biology and Drug discovery.

RESEARCH EXPERIENCE:

December-2011 –June-2015: Senior Research Fellow at Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow, U.P., India.

Febuaray-2009 –December-2011: Project Assistant on CSIR funded project entitled “Discovery and preclinical studies of new bioactive molecules (natural & semi-synthetic) & traditional preparations” at Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow, U.P., India.

PARTICIPATION IN SYMPOSIA/CONFERENCES/WORKSHOPS:

Xth Joint Annual Conference of Indian Society of Malaria and Other Communicable Diseases (ISMOCD) & Indian Association of Epidemiologists (IAE), 10th to 12th October, 2014 Goa.

XII International Conference on Vector and Vector borne diseases jointly organized by

Department of Zoology, Mohanlal Sukhadia University, Udaipur, Rajasthan India between 16th – 18th September, 2013.

5th International symposium on Drug Development for Orphan/Neglected Diseases at CSIR- Central Drug Research Institute, Lucknow, India between 26th – 28th February, 2013.

23rd National Congress of Parasitology jointly organized by The Indian Society for Parasitology and Anna University, Chennai, Tamil Nadu, India between 18th -20th November, 2011.

22nd National Congress of Parasitology, organized by Kalyani University, Kalyani Kolkata, India 30th October-1st November 2010.

Workshop attended: CDRI-BD Workshop on “Flow Cytometry Based Cell Sorting” Feb 28-Mar 1, 2012 at CDRI, Lucknow, U.P. India.

Workshop attended: CSIR- CDRI-Beckman Coulter Workshop on “Apoptosis and cell analysis” June 3-June 6, 2014 at CDRI, Lucknow, U.P. India.

MEMBERSHIP OF LEARNED SOCIETIES: Life member of The Indian Science Congress Association

SUMMARY OF RESEAECH WORKS (2011-2015 at CSIR-Central Drug Research Institute)During my PhD tenure I have opted the work related to immunological responses during pre-erythrocytic stage infection with rodent malaria parasite Plasmodium yoelii. Besides this, I have also actively engaged in other ongoing laboratory projects. The details of project undertaken by me are as follows:

1. Studies of immunological responses directed against pre-erythrocytic stage (Liver stage) infection of malaria parasite: The Pre-erythrocytic stages of malaria parasite are the least explored forms in the parasite's life cycle despite their recognition as key vaccine and drug targets. Studies with the malaria liver stages have greatly suffered due to lack of effective quantitative analysis methods for the evaluation of parasite liver-stage development and little is known about cellular immune response against pre-erythrocytic stages infection. In this view, I have standardized and validated the accuracy and quantitative analysis of liver stage parasite development by real time PCR under different experimental conditions. This method detected liver stage parasite load with as low as 50 sporozoites. The ease and speed of quantitative analysis of liver-stage development by this method provides a valuable tool for the evaluation of candidate drugs and vaccines targeting pre-erythrocytic parasite stages (Microbial Pathogenesis 2015). Besides this, I have studied the cellular immune response directed against pre-erythrocytic stage of malaria parasite (using rodent malaria model P. yoelii infection in Swiss Albino mice). For this purpose, we have performed the cytokine profiling and phenotypic characterization (density and phenotype) of liver hepatic mononuclear and spleen cells isolated from P. yoelii infected mice. Our studies suggested that parasite impaired pro-inflammatory cytokines, CD8+ T cells in HMNC and splenic cells during liver stage development. Besides this parasite also altered macrophage population in HMNC and splenic cells to successfully establish liver stage development (Manuscript under preparation). The immune response also studied using P. yoelii sporozoite induced infections in the Golden hamster model to check host immune response (hamster show nonlethal response with P. yoelii parasites). This study

has revealed significant up-regulation of pro-inflammatory cytokines in non lethal infection in hamster model, while down-regulation of pro-inflammatory cytokines in lethal infection in mice model. Taken together, our studies suggest that coordinated actions of pro-inflammatory mRNA expression are essential to control malarial infection and their attenuation leads to lethal infection (excessive parasite growth and death of animals) (Manuscript under preparation).

2. Studies of immunological responses during erythrocytic stage (Blood stage) infection of malaria parasite: To investigate the immunological basis of differences in the virulence pattern of malarial infection, we used two different strain of Plasmodium vinckei (PvAS and PvAR parasites) that caused lethal and nonlethal infection in AKR (inbred) mice and analyzed the cellular immune response directed against erythrocytic blood stage of malaria parasite. For this purpose, we have performed the cytokine profiling and phenotypic characterization (density and phenotype) of spleen cells isolated from infected mice. This study revealed that IFN-γ, TNF-αand IL-12 was up-regulated in non-lethal (PvAR) parasite while in lethal parasite (PvAS) IL-10 and IL-4 mRNA was up-regulated. Taken together, this study showed that coordinated actions of pro-inflammatory mRNA expression are essential to control lethal malarial infection and their attenuation leads to increased parasite growth and, ultimately, death of animals (Parasitology Research, 2012). Phenotypic characterization of spleen cells showed that mice infected with non-lethal parasite (PvAR) have increased population of CD4+

and CD8+ T cells, B cells, plasma cells, dendritic cells and macrophages while mice infected with lethal parasite have impaired population of CD4+ and CD8+ T cells, B cells, plasma cells, dendritic cells and macrophages. In short, this study showed that coordinated actions of pro-inflammatory cytokines and T, B, plasma & DC cells are essential to control lethal malarial infection and their attenuation leads to increased parasite growth and, ultimately, death of animals (Parasitology Research 2015).

3. Chemoprophylaxis and sporozoite immunization following repetitive live sporozoite inoculation: Inoculation with live sporozoites under prophylactic antimalarials cover (CPS-Immunization) represent an imperative way to develop sterile, reproducible, and long-term protection against malaria re-infection. However, success of the CPS-Immunization strategy is dependent on the susceptibility of parasite population to the partner drug. In this view, I have employed arteether (ART, a semi synthetic derivative of artemisinine) to explore its potential as a chemoprophylaxis candidate in CPS approach and protective efficacy of ART was systematically compared with other antimalarials (mefloquine, primaquine and azithromycin). For this purpose, sequential Plasmodium yoelii sporozoites exposure under repeated prophylactic treatment with ART, mefloquine, azithromycin or primaquine was conducted in experimental Swiss Albino mice. This study demonstrates that sporozoite administration under ART treatment confers strong protection against subsequent sporozoite infection and this protective immunization depends upon the presence and growth of liver stage parasites. Since, P. falciparum isolates across the globe are susceptible to artemisinin derivatives, CPS-ART strategy can be employed as a safe and efficacious regimen to offer a surrogate whole parasite malaria immunization strategy (Acta Parasitology 2015 Accepted). Beside this I have also studied the immunological parameter after repetitive live sporozoite inoculation under arteether (5mg/kg) chemoprophylaxis. It has been observed that pro-inflammatory cytokines (IFN-γ, TNF-α, IL-12 and iNOS), CD4+, CD8+ T cells, macrophages and DCs population was significantly elevated in protected mice, signifying their importance in protection against PE stage parasites. Besides this, I have also studied on induction of immune response in Golden hamsters after repetitive sporozoite challenge showed that five sequential inoculations

protected all hamsters against subsequent sporozoite. We have revealed that pro-inflammatory cytokines (IFN-γ, TNF-α, IL-12 and iNOS) was significantly higher in protected Golden hamster. Adaptive transfer of isolated HMNCs from protected hamsters to naïve hamsters showed 60% protection against challenge with 10,000 sporozoites and 100% protection were observed in recipient hamsters challenged with 100 or 1000 sporozoites with complete absence of 18SrRNA signal (Manuscript under preparation). 4. Identification and molecular characterization of novel antimalarial drug targets from rodent malaria parasite P. vinckei. Apart from this, I have also actively involved in molecular and biochemical characterization of putative proteins from malaria parasite P. vinckei. During the course of study, we have identified and characterized superoxide dismutase (SOD) and heme detoxification protein (HDP) from P. vinckei. PvSOD1 is cytosolic in localization and its expression is comparatively higher during trophozoite as compared to that of ring and schizont stages. Enzymatic activity of recombinant PvSOD1 was validated using conventional zymogram analyses and xanthine–xanthine oxidase system. Under optimal conditions, PvSOD1 was highly active and catalyzed the dismutation of superoxide radicals. Since, PvSOD1 plays a central role in oxidative defense mechanism (Parasitology International 2014). Heme detoxification protein (HDP) is the sole converter of heme to hemozoin in rodent parasite therefore study has been undertaken to elucidate the role of heme detoxification protein (HDP) in P. vinckei. Molecular and biochemical characterization of HDP from P. vinckei suggests that PvHDP bind with free heme and convert it into hemozoin in a concentration dependent manner and act as a bonafide heme detoxification protein (Gene 2015).

LISTS OF PUBLICATIONS:

1. Arif Jamal Siddiqui, Jyoti Bhardwaj, Manish Goyal, Kirtika Prakash, Awakash Soni, Vishwanath Tiwari and Sunil K. Puri. Assessment of Real-time method to detect liver parasite burden under different experimental conditions in mice infected with P. yoelii sporozoites. Microbial Pathogenesis (2015) 89: 35-42.

2. Arif Jamal Siddiqui, Jyoti Bhardwaj, Sunil K. Puri. mRNA expression of cytokines and its impact on outcomes after infection with lethal and nonlethal Plasmodium vinckei parasites. Parasitology Research (2012) 110(4):1517–1524.

3. Jyoti Bhardwaj, Arif Jamal Siddiqui, Manish Goyal, Kirtika Prakash, Awakash Soni, and Sunil K. Puri. Repetitive live sporozoite inoculations under Arteether chemoprophylaxis confer protection against subsequent sporozoite challenge in rodent malaria model. Acta Tropica (Accepted).

4. Jyoti Bhardwaj, Arif Jamal Siddiqui, Manish Goyal, Kirtika Prakash, Awakash Soni, Sunil K. Puri and Mrigank Srivastava. Host immune response is severely compromised during lethal Plasmodium vinckei infection. Parasitology Research (2015).

5. Kirtika Prakash, Manish Goyal, Awakash Soni, Arif Jamal Siddiqui, Jyoti Bhardwaj and Sunil K. Puri. Molecular cloning and biochemical characterization of iron superoxide

dismutase from the rodent malaria parasite Plasmodium vinckei. Parasitology International 63 (2014) 817–825.

6. Awakash Soni, Manish Goyal, Kirtika Prakash, Jyoti Bhardwaj, Arif Jamal Siddiqui and Sunil K. Puri. Cloning, expression and functional characterization of Heme Detoxification Protein (HDP) from the rodent malaria parasite Plasmodium vinckei. Gene (2015) 15;566 (1):109-19.

SKILLS AND EXPERTISE

Immunological techniques: Flow cytometery, cytotoxic T cell assay, ELISA, lymphocyte proliferation assay, estimation of various cytokines, western immunoblotting, dot blot, raising antibodies, purification of antibodies and adaptive transfer in mice.

Molecular Biology: Cloning, Real time PCR, Western Blotting, Isolation of genomic DNA, plasmid isolation, isolation of RNA, cDNA prepation, Primer designing, Polymerase Chain Reaction (PCR), Electrophoresis (Agarose , SDS-PAGE) etc

Mosquitoes breeding and rearing: Insectaries maintenance and mosquitoes breeding & rearing

Parasite Culture: In vivo maintenance of rodent malaria (P. yoelii, P .vinckei) parasites. Antimalarial Screening: In vivo Antimalarial screening of chemotherapeutic agents

(synthetic compounds and natural products) in rodent and monkey models. Microscopy: fluorescence and light microscopy. Animal handling: Mice, Hamster, Gerbil, Rabbit and Monkey (expertise intravenous,

intraperitoneal, intradermal, subcutaneous and intraorbital routes). Apoptosis: - FITC and Annexin–V binding assay, Estimation of reactive oxygen species

and phagocytosis assay Computer skills: Flow-cytometry data analysis using Flow-Jo software, MS office, MS

excel, Endnote referencing, MS DOS, Paint, Adobe Photoshop etc.

REFEREES:

1. Dr. S.K. Puri, PhDChief ScientistParasitology DivisionCSIR-Central Drug Research Institute, Lucknow, India.Mobile: +91- 9919566792Tel. No. +91-522-2772453E-mail: sk_puri@cdri.res.in skpuricdri@yahoo.co.in

2. Dr. Shailja Bhattacharya Chief Scientist, Division of Parasitology, CSIR-Central Drug

Research Institute, Lucknow, Pin-226 031, U.P., INDIA Mob. No. +91-9415010721 Tel. No. +91-522-2772455 E-mail: shailja_bhattacharya@cdri.res.in