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Training Programme

TRAINING PROGRAMME IN MOLECULAR BIOLOGY AND IMMUNOLOGY OF LEPROSY

Schieffelin Institute of Health – Research and Leprosy Centre (SIH-R & LC) located near Vellore  in Tamil Nadu, was established in 1955 to care for the leprosy patients in and around Vellore and to set up a research institution to probe into the multifaceted problems of leprosy. Dr. Robert Cochrane was the moving force behind this venture. Over the past decades SIH-R & LC has grown into a centre of excellence as tertiary care centre for leprosy services, International Leprosy Training Centre and a State of the Art Leprosy Research Centre.

Molecular Biology Laboratory was started in the year 1999 to perform basic science research in leprosy in order to understand the genomic and molecular constituents of Mycobacterium leprae and to utilize the knowledge to identify effective diagnostic, prophylactic and therapeutic measures to combat leprosy. Various studies have been pursued in collaboration with “The American Leprosy Missions (ALM), USA”, “Infectious Disease Research Institute (IDRI – Seattle, Washington, USA)” and “The Leprosy Mission Trust India”.

Apart from the regular research grants from SIH-R & LC, the laboratory was extramurally funded by ALM, TLM, IDRI, National Institute of Health (NIH, Maryland, USA) and Indian Council of Medical Research (ICMR, New Delhi, India). The laboratory has state of art facilities for Polymerase Chain Reaction (PCR) based gene amplification and detection systems for mycobacterial genome analysis, chemiluminescence assays, enzyme linked immunosorbent assays, T cell culture systems, microbial culture facility, fully equipped animal house facility for mice as a model for experiments with facilities for irradiation and thymectomy. A central library facility is available to access various books and scientific journals. The lab is facilitated with computers which are connected with high speed wifi internet.

The laboratory has worked broadly in the areas of:

  1. Identifying molecular markers for early diagnosis of leprosy.

  2. Effectiveness of alternative drug regimens for treating drug resistance in leprosy.

  3. In situcytokine mRNA expression profiling during early stages of  leprae infection.

  4. Molecular Epidemiology studies on nasal carriage and mucosal immunity.

  5. Variable Number Tandem Repeat Analysis of Mycobacterium leprae genome to understand molecular basis for the variance in strain types.

  6. Environmental Mycobacterium leprae.

  7. Immunological Studies on Type I and Type II Reactions in leprosy.

Being a DSIR (Department of Scientific and Industrial Research – Ministry of Science & Technology, Government of India) recognized research laboratory, the unit undertakes training programs and student projects in Molecular Biology and Immunology aspects of leprosy apart from the regular research projects.

The details of the programs are as follows:

  1. Short-term Training Programme & Projects

Duration:  2 – 3 Months (Maximum Period)

 Training Modules:

  1. Basics of Molecular Biology and Immunology in the context of leprosy.

  2. PCR based amplification of Human and  leprae specific gene targets and detection through Agarose gel electrophoresis.

  3. ELISA and chemiluminescence assays for detection of specific immune markers

  4. Western and Northern Blotting.

  5. Research Methodology and Data Analysis.

            Fees:  INR 6,000/- (For the entire training program)

             Eligibility: Students who are currently pursuing

  • B.Sc in Life Sciences.

  • M.Sc. in Life Sciences.

  • B.Tech Biotechnology

  • Integrated M. Sc in Life Sciences.

Food & Accommodation:

In the Guest House: As per the standard Guest house tariffs.

(Kindly Contact Mrs. Daya at the Guest House No: +91 0416 2274227  Extn:  2500  for the tariffs).                                                      

In the Students’ Hostel: (Negotiable)

Rs.50/- per day for accommodation

Rs. 100-120/- per day for food.

  1. Dissertation Projects

Duration:  6 Months – 1 Year (Maximum Period)

Project Description:

A research project will be assigned to the students, which needs to be accomplished in a period of 6 months – 1 year as per the requirements of the college/university from where the student is enrolled on a full time basis. Admission of students in this category is subject to their full time enrollment in a college/university and their curriculum comprises a full time dissertation project*. The projects will be in thematic areas of current Molecular Biology & Immunology aspects of Leprosy.

Fee: Rs. 15,000/- (For the entire project)

Eligibility: Students who are currently pursuing

  • M.Sc. in Life Sciences.

  • B.Tech Biotechnology

  • Integrated M. Sc in Life Sciences.

Note: Students in this category need to submit a letter from their college/university that the project work is a part of their course curriculum.

Food & Accommodation: same as above.

Key Activities of the Department:

  1. Development of Molecular Biology and Immunology based diagnostic tools for early diagnosis of leprosy.

  2. Immunopathology of Reactional States in Leprosy.

  3. Development of molecular epidemiological tools to understand focal transmission of leprosy in the endemic areas.

  4. To identify effective diagnostic and therapeutic methods for nerve function impairment in leprosy.

Research Publications:

Published:

  1. Das M, Chaitanya VS, Kanmani K, Rajan L, Ebenezer M. Genomic diversity in Mycobacterium leprae isolates from leprosy cases in South India. Infection, Genetics and Evolution. 2016 Nov 30;45:285-9.

  2. Chaitanya VS, Das M, Bhat P & Ebenezer M. Computational Modeling of Dapsone interaction with Dihydropteroate synthase in Mycobacterium leprae; Insights into molecular basis of dapsone resistance in leprosy. J Cell Biochem. 2015 Apr 2. doi: 10.1002/jcb.25180. [Epub ahead of print].

  3. Chaitanya VS, Das M, Eisenbach TL, Amoako A, Rajan L, Horo I & Ebenezer M. Mycobacterium leprae specific genomic target in the promoter region of probable 4-[alpha]-glucanotransferase (ML1545) gene with potential sensitivity for PCR based diagnosis of leprosy. International Journal of Mycobacteriology, Available online 16 February 2016, ISSN 2212-5531, http://dx.doi.org/10.1016/j.ijmyco.2016.01.002.

In Press:

  1. Evaluation of anti-bacterial activity of Rifapentine, Clarithromycin, Minocycline, Moxifloxacin, Ofloxacin and their combinations in murine model of Rifampicin Resistant Leprosy.Indian Journal of Leprosy – 2016

  2. Evaluation of new Antibacterial Drug Combinations in Murine Model of leprosy to identify short duration alternative to Standard WHO regimen of Multibacillary – Multidrug Therapy.Indian Journal of Leprosy – 2016

  3. Analysis of a novel multiplex PCR assay as a sensitive tool for diagnosis of indeterminate and tuberculoid forms of leprosy – International Journal of Mycobacteriology– 2016

Approved Projects:

  1. To identify Mycobacterium leprae associated transcriptomic and host immune signatures that aid as early signals for determination of reactional states in leprosy.

Funding Body: The Leprosy Research Initiative – Netherlands

Project Duration:   Four Years (2016-2020)

Expected Outcome:       

  1. Determination of leprae associated transcriptomic signatures that demonstrate significant difference in expression in the skin lesions of reactional (type I and type II) leprosy cases when compared to those without reactions (Identified through whole  leprae transcriptome analysis).

  2. Identification of blood- as well as lesional skin -derived host mRNA expression levels of genes that encode active immune molecules that significantly vary between the leprosy cases in type I and type II reactions and those not in reactions.

  3. Quantitation of specific immune molecules in cell culture supernatants of Whole Blood assays (WBA) using Bio-Plex-suspension array technology to differentiate type I and type II reactions from non reactional cases.

  4. Further validation of the above identified differentially expressing mRNAs and cyto-/chemokines in a longitudinal design (before, during and after reaction in the same patient) to select a final set of signatures that can be used as potential analytes for developing an early diagnostic test for type I and type II reactions in leprosy. (Using Multiplex-Real-time PCR assays and Up-converting Phosphor- Lateral Flow assays (UCP-LFA)).

Diagnostic Activities:

  1. Screening for Drug Resistance in Leprosy:Molecular Biology Laboratory, SIH-R&LC, Karigiri was recognized as one of the apex laboratories to screen for drug resistance in leprosy in India by ICMR. In this context, drug resistance screening of leprosy patients was initiated with appropriate processing fee (Rs. 2000/- per sample). The investigations included PCR based amplification of genes gyrA, folp and rpoB. This is followed by purification of PCR amplicons and then DNA sequencing to detect mutations that confer resistance.

The following biopsies were received from relapsed/non-respondent leprosy patients, processed and sent for DNA Sequencing.

Sl. No.

Name of the Institute

Number of Skin Biopsies Received/Results Reported

1

Father Muller Foundation, Mangalore

14

2

St. Johns Hospital, Bangalore

1

3

SIH-R&LC Karigiri

30

4

JIPMER– Pondicherry

2

5

Govt. Medical College -Trivandrum

18

6

Foundation for Medical Research – Mumbai

1

7

Christian Medical College

1

8

JSS Medical College Mysore

1

  1. PCR based diagnosis of leprosy: PCR based diagnosis of leprosy was also being continued with two biopsies from suspected leprosy patients from the department of Dermatology, CMC, Vellore. This was also being carried out with a fee of Rs. 500/- per sample. The investigations include mycobacterial DNA extraction from biopsy samples and PCR for ML1545 and rlep gene targets.

Sl. No.

Name of the Institute

Number of Skin Biopsies Received

PCR for ML1545 and rlep gene targets

Results Dispatched

1

Department Dermatology – CMC, Vellore

14

Completed

Yes

  Training Activities: Being a Department of Science and Industrial Research (DSIR) recognized laboratory, training in short term projects and curricular dissertation projects has also been initiated in the lab. Two student projects were accomplished which are as below:

Ms. Madhusmita Das- Research assistant in the lab has been enrolled with Martin Luther Christian University in Shillong for the PhD program on the topic: Immuno-epidemiological, clinical and molecular biological investigations on transmission of leprosy.2015-2017

Sl. No.

Name of the Candidate & Institution

Title of the Project

Duration

1

Ms. Tiffany Lynn Eisenbach – St. Olaf College, Minnesota, USA

Detection of rlep gene target of Mycobacterium leprae from clinical Isolates of leprosy patients, their familial contacts & controls in the leprosy endemic area in India, using PCR.

11th Nov 2014 – 16th Dec 2014

2

Ms. Angela Amoako– St. Olaf College, Minnesota, USA

A pilot study to identify the potential of two novel Mycobacterium leprae specific overexpressing gene targets ML1545 and ML0307 as tools of sensitivity for PCR-based diagnosis of Leprosy with rlep gene target as the control.

11th Nov 2014 – 16th Dec 2014

3

Ms. Luz Cuello – St. Olaf College, Minnesota, USA

A study to determine the utility of Multiplex PCR using Mycobacterium leprae specific gene targets, as a tool for confirmatory diagnosis of indeterminate and tuberculoid leprosy.

11th Nov 2015 – 16th Dec 2015

4

Mr. Hunter Lin – St. Olaf College, Minnesota, USA

Development of a prototype for quantification of color signals generated by lateral flow assay strips designed to assess cytokine profiles in Leprosy Type 1 Reactions

11th Nov 2015 – 16th Dec 2015

5

Ms. Malathi Manuel – University of Madras, Chennai, India

Internship in Molecular Biology and Immunology of Leprosy

25th May 2015 – 25th June 2015

6

Ms.  Shamili Neelagandan – University of Madras, Chennai, India

Internship in Molecular Biology and Immunology of Leprosy

25th May 2015 – 25th June 2015

7

Mr. Sudharson – Anna University – Chennai

A study to investigate gene expression profiles of IFN-Gamma, E-Selectin, TLR-2 and TLR-10 and serum levels of IFN-γ during reactional states of leprosy so as to determine their utility as prognostic markers.

January –May 2016

8

Ms. Kanmani –  Anna University – Chennai

A study to investigate the distribution of molecular types of Mycobacterium leprae based on single nucleotide polymorphisms from leprosy endemic areas in Tamil Nadu.

January –May 2016

9

Ms. Pavitrra-  Anna University – Chennai

A study to determine the loss in viability at different time points of drug dosage in mice treated with alternative combinations of antibacterial drugs against Mycobacterium leprae.

January –May 2016

10

Mr. Shansank Swain

Evaluation of Dapsone Congeners for their bactericidal activity on dapsone resistant M. leprae in a Murine model

August –September 2016

11

Mr. Mukul Sharma

Expression profiling of genes involved in DNA repair mechanisms in Mycobacterium leprae.

August –October 2016

12

Ms. Megan Bristow – St. Olaf College, Minnesota, USA

To elucidate the possible association of efflux pump gene expression with drug resistance in Dapsone and Rifampicin Resistant Mycobacterium leprae.

November-December 2016

13

Ms. Rebecca de Guzman  Ferrer   – St. Olaf College, Minnesota, USA

A study to investigate the distribution of molecular types of Mycobacterium leprae based on real time PCR and High Resolution melt Analysis from leprosy endemic areas in Tamil Nadu.

November-December 2016