Journal of Postgraduate Medicine Medknow Publications and Staff Society of Seth GS Medical College and KEM Hospital, Mumbai, India ISSN: 0022-3859 EISSN: 0972-2823 Vol. 46, Num. 4, 2000, pp. 297-302

Journal of Postgraduate Medicine, Vol. 46, No. 4, October-December, 2000, pp. 297-302

Looking Back

National Institute of Virology

20A, Dr Ambedkar Road, Post Box No.1 1, Pune - 411001, India.

Code Number: jp00099

The National Institute of Virology is one of the major Institutes of the Indian Council of Medical Research (ICMR). It was established at Pune, Maharashtra State in 1952 as Virus Research Centre (VRC) under the auspices of the ICMR and the Rockefeller Foundation (RF), USA. It was an outcome of the global programme of the RF for investigating the Arthropod Borne viruses. Since the studies on arboviruses and their arthropod vectors involve most of the basic principles and techniques of general virology, entomology and zoology, these viruses were also considered to be an ideal group, to begin with, for intensive training and research in virology in our country. The RF withdrew its support in 1967 and since then the Institute is entirely funded by the ICMR.

The research activities of the centre were made more meaningful and self reliant by organizing new areas of research, such as Cell repository, Electron microscopy, Rickettsioses, Hepatitis, Influenza and related viruses, Clinical virology, Biochemistry, Virus registry, and Biostatistics. The research activities of the Institute are coordinated by a Scientific Advisory Committee (SAC) consisting of eminent scientists.

By the 70s the Institute had developed deep scientific roots, nurtured through the sustained efforts of many dedicated workers. With the expertise in virological training and research and emphasis on selfreliance, the centre was well prepared to undertake full responsibility as a National Institute. On the recommendation of the SAC, the VRC acquired its status of national importance and was renamed as National Institute of Virology (NIV) in 1978. Subsequently studies on Acquired Immune Deficiency Syndrome (AIDS), Rotavirus gastroenteritis, Acute haemorrhagic conjunctivitis, Rabies, Herpes sirnplex, Buffalo pox, Measles, and Poliomyelitis were also initiated.

A Microbial Containment Complex (MCC) having P3 biosafety levels for handling microorganisms of highly infectious nature is being established at Pashan, 11 km off the main laboratory at Pune. This laboratory will provide National Containment facility for safe handling of highly hazardous pathogens.

The Institute was designated as one of the collaborating laboratories of the World Health Organization (WHO) in 1967 and it started functioning as the regional centre of the WHO for SouthEast Asia for arbovirus studies from 1969. Since 1974, it has been functioning as a WHO collaborating centre for arbovirus reference and research. In 1995 it has been redesignated as the WHO Collaborating Centre for Arbovirus and Haemorrhagic Fever Reference and Research and Rapid Diagnosis of Viral Diseases. NIV is also the National Centre for Hepatitis and Influenza. The field unit of NIV at Bangalore is one of the centres under National Polio Surveillance Program conducting surveillance of acute flaccid paralysis cases from Kamataka as a part of Global Polio Eradication Prograrnme of the WHO SouthEast Asia region since 1997.

Priority Areas

1. Vector borne viruses Epidemiology, Pathogenesis, Transmission cycle, Bio-ecology of vectors and vertebrate hosts if any, Vector Control, Pesticide Resistance, Insect genetics.
2. Development of Immunobiologicals Development and evaluation of both new and established antiviral vaccines, development and evaluation of rapid diagnostic tools.
3. Development of maximum microbial containment facilities for investigating highly pathogenic viruses.
4. Epidemiology, molecular biology, immunology and ultra structural studies of viruses of public health importance viz. Japanese Encephalitis, Dengue, Hepatitis, Measles, Influenza, Rota, etc.

Scientific Activities

The Institute has conducted in depth studies on various aspects of many arboviral diseases; epidemiology, ecology, pathogenesis, transmission cycles, bioecology of arthropod and vertebrate hosts, vector control, pesticide resistance and development of vaccines. These arboviruses are viz. Kyasanur Forest Disease (KFD), Japanese encephalitis (JE), West Nile (WN), Dengue (DEN), Chikungunya (CIEK), Sandlly Fever (SF), African Horse Sickness (AHS) etc. It has also conducted similar studies on other viral diseases of public health importance, viz. Hepatitis, Influenza, AIDS, Rota, Buffalo pox, Measles etc.

Epidemiological, Virological, Entomological, and Zoological Studies

The Institute has investigated so far over 225 epidemics of suspected viral aetiology in the country, including the Andaman and Nicobar islands; and was able to isolate and identify causative agents in many of these epidemics. It has conducted numerous serological surveys in different parts of the country in order to determine the distribution and prevalence of viruses in the country. It has an invaluable collection of nearly 250,000 serum samples, most of which are human sera collected during epidemics and serological surveys from all over India. Serological surveys of a variety of animals and birds have also been carried out. In addition, numerous entomological and zoological surveys have been carried out in different parts of India, including the Himalayan regions. The virus registry of NIV has 502 virus strains belonging to 12 families, which also include some ungrouped virus strains.

Studies on the involvement of blood sucking arthropods and nonhuman vertebrates in the natural cycle of arthropodborne viruses, resulted in extensive studies on medical entomology and zoology. The NIV has carried out extensive faunistic, taxonomic, bioecologic and disease relationship studies on blood sucking insects and acarines, birds and mammals in different parts of the country. A large number of new species of various blood sucking arthropods, viz. one species of mosquito, two species of sandflies, nine species of sucking lice, two species of fleas, three species of bugs, 18 species of ticks, 63 species of trombiculid mites and one species of chalcid have been described. One new species, each of rodent, bat and bird, has also been described by NIV scientists.

Vaccines

Studies conducted by NIV on the unravelling of the natural history of KFD and the development of vaccine against it, is considered to be one of the classic works in Arbovirology.

The Institute has developed a formalised inactivated chick embryo tissue culture vaccine against KFD. This was the first ever viral vaccine developed and produced entirely with Indian expertise. The technology was transferred to the Karnataka Public Health Department, which has manufactured thousands of doses of vaccine in its laboratory at Shimoga and administered the vaccine in the population at risk.

Attempts to develop vaccine against Japanese encephalitis are continued at NIV. Peptide fragments of proteins can induce immune response. This principle can be used to produce synthetic peptide vaccines. Synthetic peptides from JE virus envelope protein inducing T helper activity and antibody response, have been identified by computer programmes based on physicochemical parameters of amine acids.

The Institute has conducted vaccine trials with inactivated mouse brain derived JE vaccine prepared at Central Research Institute, Kasauli. Evaluation of rabies (HDCV), HBV (Engerix B) and measles vaccines has been carried out by the NIV. The field unit of the NIV at Bangalore has undertaken potency testing of field samples of oral polio vaccine in Karnataka state since 1992.

Vector Control

The Institute maintains laboratory colonies of a number of species of mosquito, tick, flea and trombiculid mites for experimental use such as transmission, insecticide resistance and repellent studies. As a part of the control of viral diseases, studies have been carried out in the laboratory as well as in the field, on the control of arthropods of medical importance by using insecticides, repellents and biocides. The important field studies include those in the KFD area in Karnataka state using acaricides (Lindane) and repellents (Deet and Mylol) against ticks; control of Aedes aegypti in Pune city and Dehu Road, by using Abate and other source reduction methods. In addition, many vector species including mosquitoes, ticks, sandflies, mites and fleas from many parts of the country, have been screened to determine the insecticide resistance/susceptibility status. The information thus obtained has been passed on to the public health workers.

Tissue Culture and Cell Biology

Cell cultures derived from organs and tissues of various animals have provided an elegant system for virological studies. They have replaced the use of expensive laboratory animals to a great extent.

The world's first mosquito cell line, popularly known as Singh's cell fine, was established at the NIV in the year 1967. It is being used for studies on arboviruses. A number of cell fines of invertebrate and vertebrate origin such as VRC 4 of human amnion origin and MRK 90 of Macaca radiata kidney origin have been developed. Developmental, karyological, histochemical, biochemical characterisation and virus susceptibility studies have been carried out on established and newly developed cell lines.

Recently eight cell lines from four different lepidopteran insects (moths) have been developed. Such cell fines provide a system for the growth of baculoviruses. Baculoviruses are being increasingly used to synthesize recombinant proteins. The cell lines developed at NIV promote significantly higher growth of baculoviruses. Naturally occurring baculoviruses can be grown and used as biopesticides. Development of fish cell culture is undertaken by the Institute on a request from the department of fisheries, Government of India, for use in the investigation of a haemorrhagic disease affecting economically important fishes of India. Two cell lines from fish organs have been developed.

The NIV maintains over 150 cell fines in its cell repository. It has been supplying these cell fines to different institutes in the country and has thus rendered yeoman's service for other scientists working in different fields of biomedical science.

Electron Microscopy

The major objective of the Electron Microscope division is to provide support to the activities in diagnostic virology. In addition, the division is conducting studies on the cellular morphogenesis patterns of various viruses.

Commissioning the country's first Digital Cryo Electron Microscope

The Institute successfully installed and commissioned a high resolution stateof-art digital Cryo Transmission Electron Microscope complete with online digital imaging systems and equipped with electron tomography setup. The Microscope has GATAN cryo transfer work station for rapid ultra low temperature freezing of biological specimens and is equipped with a computer controlled cryo stage that maintains ultra low temperature during specimen observation. The major applications of this microscope will be in studying virus pathogenesis, Cryo Electron Microscopy of virus structure including 31) reconstructions based on electron tomography and rapid diagnosis. Application. In other areas of infectious diseases is also possible and this Facility at NIV is created with an aim of becoming a National Centre for Excellence in Biological research with an emphasis on virology and infectious diseases.

Development of newer technologies for rapid viral diagnosis

1. Enzyme Linked Immunosorbent Assay (ELISA) kits: Rapid diagnosis of viral infections assumes importance for appropriate symptomatic treatment. Many diagnostic kits need to be imported which are expensive. NIV has developed several diagnostic kits indigenously. ELISA have been developed for detection of hepatitis B infection (hepatitis B surface antigen, HBsAg) from blood specimen and detection of rotaviruses from stool specimen. However, detection of viruses from clinical specimen is not always possible and diagnosis can be based on other markers. IgM antibody appears early during infection and detection of virus specific IgM antibody correlates with recent infection. NIV has developed IgM detection ELISA tests for rapid diagnosis of Japanese encephalitis, Dengue, West Nile, Hepatitis A, and Measles virus infections.
2. Monoclonal Antibodies (MAbs): The development of MAbs has a major impact on diagnostic virology and has led to the development of numerous commercial viral diagnostic kits. The use of MAbs provides highly specific results. Studies on mouse MAbs have been undertaken since 1985 at NIV. A large panel of MAbs have been developed against JE, WN, DEN, Chikungunya viruses. The MAbs have also been used for strain analysis and epitope mapping.
3. Polymerase chain reaction (PCR): The discovery of this new technique in recent years, has revolutionized the field of viral diagnosis. The PCR has important applications in instances where traditional viral isolation techniques are difficult to perform. The NIV has developed PCR based tests for Hepatitis B, C and E Viruses to determine the aetiology and to generate epidemiological data specially, on strain variation. PCR amplification and subsequent nucleotide sequencing has opened a new field of molecular epidemiology. Detection of precore mutants of HBV with clinical and treatment implications, geographic distribution of HCV, BEV, HGV and a novel DNA virus TTV, were possible because of modem technologies. By fully utilizing these technologies, NIV has taken a leadership role in the field of virology in the country.
4. Facilities for other advanced studies: In addition to above mentioned techniques NIV has excellent infrastructure for many advance studies. Animal house and tissue culture facilities for virus isolation, ultracentrifuges for virus purification, electron microscopes for ultrastructural studies, several ELISA work stations for serological tests, BPLC and electrophoresis facilities for protein purification and protein analysis; an immunology division for cell mediated immune responses against viral infection, facilities for peptide synthesis and flow cytometry, etc.
5. Mosquito inoculation: Mosquito inoculation technique has been utilized routinely for the detection of certain Flavi viruses (JE, DEN) from field caught adult mosquitoes. Besides, intracerebral inoculation of mosquito larvae has been developed for rapid detection of JE virus. These techniques have been found to be sensitive and simpler than the conventional methods of mouse and tissue culture inoculation.

Infrastructural Facilities

The NIV maintains a well designed facility for the upkeep and production of high quality laboratory animals including nonhuman primates. The Institute has an good reference collections of Indian ticks, sandflies, fleas, sucking lice, trombiculid mites, birds and mammals in its museum. The Institute has several sophisticated viral equipments and cryopreservation facilities; cold rooms, freezer rooms, deep freezers, liquid nitrogen, etc. The Institute's library is considered to be one of the very good scientific libraries in the country. Scientists and technical personnel from various Institutes in India and abroad are trained at NIV for varying periods of time.

Field Units

The NIV has been maintaining some field units from time to time in different parts of the country, to study regional viral diseases. A field station functioned at the Christian Medical College, Vellore, Tamil Nadu from 1955 to 1970, and this unit was intended to study the epidemiology and ecology of J13 and DEN viruses in the region. This unit was subsequently shifted to Bangalore in 1971 to study DEN and JE in the region. Another field unit functioned at Sagar, Shimoga District, Karnataka from 1956 to 1976 to study exclusively KFD. With the outbreaks of JE in Kolar area, Karnataka State, a field unit was set up there in 1980 which continued there up to 1992, and was later shifted to Mysore to study arboviral problem in Mysore and Mandya Districts. A field unit was also set up in Gorakhpur, Uttar Pradesh (1988-2000) to study ecology of JE vectors in the region.

Academic Activities

Postgraduate degrees / Publications

The Institute is recognized for postgraduate studies by many universities. So far a total of 84 scientists have received postgraduate degrees; M.Sc. (28), Ph.D. (47), M.D. (3), M.Phil (5), M.Lib. (1). About 1020 scientific articles have been published in national and international journals by NIV Scientists. The NIV has published booklets on JE, KFD DEN, Hepatitis, Influenza, gastroenteritis, Rabies, Handbook on NIV, technical brochure on Development of lepidopteran cell lines for baculovirus studies and monographs on Indian ticks and lice. It has also published popular books for educational purpose.

Honours / Awards

National honours and awards have been received by 29 scientists of NIV.

Teaching and Training

Several scientists of NIV have been abroad for advanced studies and training to improve and update their scientific knowledge. They have also been going abroad to participate in international conferences and meetings.

Scientists and technical personnel from various institutes in India and abroad are trained in the NIV for varying periods of time. The Institute conducts one year course leading to Diploma in Medical Virology of the University of Pune. The scientists actively participate in teaching and training programmes of the University of Pune. The expertise generated at NIV has formed the basis for the development of more centres for virological and related studies such as the National Centre for Cell Science, National AIDS Research Institute at Pune and Vaccine Development Laboratory at Shimoga.

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