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					                                  SAMPLE PROPOSAL 1

                      Out Line of the Proposed Topic of Research

Name of the candidate         :             ******************
ID No.                        :             200*PHXF***
Place of Research Work &
Organisation                  :             Name, City

Proposed Supervisor Details
Name                          :             Dr. ******
Qualification                 :             Ph. D.
Designation                   :             Only Designation
Organisation                  :             Name, City

Proposed Topic of Research
Evaluation of Accounting Concepts in Indian Industry

Objective of the Proposed Research

The research focuses on improving the electroplating process by taking into account the interest
of various stakeholders such as manufacturer, designer, end user and the environment by
understanding it in a more systematic manner. The main objectives of the proposed work to be
carried out are as follows:
           1. Structural and attributes based system modeling.
           2. Coding, evaluation and selection of optimum system for different applications.
           3. Development of reliability and quality models and performance indices of total
           4. Modeling and analysis of process sub systems up to component level.
           5. System analysis for design and performance improvement.
           6. Design for human aspect and environment.
           7. Failure analysis and Total quality management of the system.

           8. Performance-evaluation, analysis and modification through experiments on
                electroplating set-up.
Background of the Proposed Research
The demand of better surface quality of products by designer, manufacturer, supplier, and end
user and the need for optimum surface treatment to aim at better surface quality to face the
unknown situation can never be over emphasized. Development of mathematical model and
analytical method to generate optimum surface treatment process at the conceptual stage for a
given application is a great challenge for the surface treatment engineers as well as for
manufacturer. The method which gives the better understanding of the surface treatment
parameters, its linkage with each other, and critical and sensitive parameters should be accurate
and comprehensive in nature. It is well established that the overall performance does not depend
only on the process, but also on other factors like human aspect, equipment, control, environment
etc. The identification of different parameter in surface treatment process and a comprehensive
model is proposed. The factors like cost, quality, environment, etc play an important role in the
performance of the surface treatment methodology. Thus all these factor are to be considered
together with their interdependence on each other. The total electroplating surface treatment
process can be defined as a system, when product, process and other subsystems along with
different interactions are considered together.
Electroplating is considered to be frequently used surface treatment process [1]. The
electroplating set up is also available in the work shop of BITS Pilani, Goa campus for
experimentation and analysis as shown in figure 1.

Literature Review of Research Topic
There is a wide variety of surface treatment methods available for applications on any products
which meets the requirement of user, supplier or manufacturer and make the product more useful
to them. Large numbers of such methods are available and continually increasing as market has
become more sophisticated and more demanding. However, over the past decades there has been
a changing pattern in electroplating requirements in terms of simplicity, economic operation,
durability, wear resistance, aesthetic, formation of highly sophisticated parts etc. This has
stimulated research and development interest to understand the technology. Quality, price,

      aesthetic, flexibility, reliability etc. as explained by Dowey and Matthews [2], Chan and Wu [3]
      and Flott [4, 5] for industrial organizations, including manufacturers and users of surface
      coatings are increasingly seeking to ensure conformance to target specifications and low
      variance; using measured variables in a product or process that can be used to assess. In our work
      it is proposed to discuss how a non-normal physical phenomenon can be accepted as a quality

                                                                               D.C. Source

Plating Tank

                                                                  -                         (Work Piece)
   Anode (Plating
   Metal)                              Figure 1: Electroplating setup up
      Chan [6] has applied the case based reasoning- a well known Artificial Intelligence (AI)
      technique for electroplating industries, which has already proven its effectiveness in numerous
      industries. This technique ensure sufficient profit margin for electroplating manufacturer by
      grasping the coating weight of electroplating component accurately so that salespersons can
      make sure their quotation prices cover the precious metal cost [7]. Apart from quotation accuracy
      and responsiveness are also a critical competitive edge in electroplating industry. AI is also used
      by Chan and Huang [8] for management of the pollution. In this study the researcher has not
      identified the other factor like human aspect, aesthetic etc. which also affect their objective
      function. Related to this, a Fuzzy Logic (FL) concept is also applied by Lin and Lin[9], which
      ensure to convert knowledge from experienced staff; simulate the ‘mind-set’ of decision maker
      in solving problem through acquisition of specific knowledge and experience; and build up self-
      learning characteristics. It is also evident that in the selected area of surface treatment process

and product there is a need for enhancement in the method of data availability, reliability,
methodology validation and system complexity. There is a scope of research work for the
selection of critical and sensitive parameter and finding the interdependence of these for
optimizing the combination of it. This innovative concept differentiates it from previous research
work. It will be verified by conducting appropriate experiments on the electroplating set up with
us. Further, it can also be suggested that it is very difficult and not practical to develop a pure
robust system which is unaffected by the environmental factor like human error, atmospheric
condition etc. Applying some subjective guiding rules significant improvement in the
performance of system in the early learning stage can be found out. Wong et al. [10] emphasized
on decision support system- a management information system for meeting the specific
requirement of industries for planning and job assignment, but scheduling is missing in his work.
The optimization technique developed by G. Taguchi called as Taguchi method is also applied
by number of authors as Flott [11], Mascio and Barton [12], Gaitonde et al. [13], Nian et al. [14],
Singh and kumar [15], which identify setting of the adjustable factors that makes the process less
sensitive to the variation. Here the researcher have not applied the development of unified frame
work for studying and developing robust process optimization and process control techniques.
Thus a robust process optimization technique can be suggested, which utilizes all available
measurements of raw material qualities at the start of each process. By this technique a reduction
of variability due to variation in raw material qualities, compared to ordinary robust process
optimization is achieved. It also deals with the analysis of the process parameters such as
influential factors, on the objective function considered based on Taguchi’s experimental design
methods. Taguchi’s tools such as orthogonal array, signal-to-noise ratio, factor effect analysis,
analysis of Variance (ANOVA) etc. have been used for this purpose and an optimal condition
has been found out. The optimum parameter combination was obtained by using the analysis of
signal-to-noise (S/N) ratio. The results have been compared with the results obtained by the
conventional experiment. The Taguchi method is suitable to solve the stated problem with
minimum number of trials as compared with a full factorial design. There is a scope of extension
of Taguchi method of design of experiment (DOE), for surface treatment system, finding its
significant factor and effects of interaction among sub system and sub sub system with its
different attributes. The gap also exits in the area of application artificial neural network (ANN)
using Taguchi method which can be implemented for minimizing objective functions relevant to

the surface treatment system. The orthogonal array and the results of simulation can be used as
training data of ANN. The results of analysis to validate the proposed design method can be
presented. Also the application of the Taguchi method with fuzzy logic for optimizing the
multiple objective performance characteristics, i.e., Multiple Analysis of Variance (MANOVA)
can be suggested. Experimental results can also be presented to demonstrate the effectiveness of
this approach. There is also a gap in present literature regarding displaying control quality
graphically with stability and performance qualities as the two axes can be used to highlight the
trade-off between these two characteristics. Some technique can be proposed to find out the
degree to which system inputs and outputs depart from their nominal values, which may prove
useful in conveying control quality in a common language understood by designer, engineers,
operators and management. However, the limitation and disadvantage of Taguchi method is
presented by Parks [16]. It states that HPD (Holistic Probabilistic Design), which holistically
treats Stochastic Optimization and thus includes comprehensively and rigorously addressing
robustness and not by Taguchi robust design.
Mohan et al. [17, 18] have presented a mathematical model using the graph theory and a matrix
method is also given for evaluation of performance of coal based Steam power plant [19]. A top
down approach for complete analysis of any system is also given. Also the same theory is
extended to determining the maintenance critically index and then suggesting a suitable
maintenance strategy.     This method can be extended further for modeling the complete
electroplating surface treatment process by considering system and sub system and representing
them by a block diagram and also into a mathematical model for analysis and synthesis of
process and product from different perspective.
Gandhi and Agrawal [20-23] presented a method of failure mode and effect analysis (FMEA),
reliability and wear analysis system based on digraph and matrix approach. This takes into
account the interrelation of all sub system and attributes of system and represent then into matrix
form for detailed computer analysis. Reliability index is presented which provides the user
directions for minimizing the failure caused leading to improvement of the system reliability.
Khan et al. [24] propose the Strength Weakness Opportunity and Threats (SWOT) function
deployment approach based on a matrix for a maintenance system. This approach would
ultimately lead to higher productivity with requisite amount of product quality. This approach
can also be extended for surface treatment industries for its failure, reliability and SWOT

analysis. A matrix and indices can also be proposed for this, which helps in faster storing,
retrieval and processing of data for its computer analysis.
Grover et al. [25] identifies factors responsible for the Total Quality Management (TQM) and
develops a mathematical model from interacting factors using a digraph approach, which is
useful for computer processing. Venkataswamy and Agrawal [26-28] developed a structural
model for an automobile and the evaluation of various performance measures in totality based on
the structure of the automobile. It also presents a coding and classification scheme for different
components of automobile vehicle. Further Bhangale et al. [29], Ahluwalia et al. [30] has given a
methodology for attribute based specification, comparison and selection of a robot and for a
Roller bearing respectively. The methodology is based on Multiple Attribute Decision Making
Method (MADM) called TOPSIS (Technique for Order Preference by Similarity to Ideal
Solution) which can be used for selecting an appropriate design of product or process system in
totality. Al-Hakim et al. [31] also uses graph theory to represent a product and define the
relationships between its components. Thus, it is evident from critical literature review that there
is a scope for extension of work to carry out in-depth study of the total electroplating surface
treatment system and aspect of TQM evaluation and its interrelation with other attributes.

Gap in existing Research
It is clear from the above literature review that no body has considered surface treatment system
as a whole, although some of the authors have consided only some sub systems separately like
work piece material, coating material etc.[32, 33], but not all the factors and their interactions
together. The analysis of the Electroplating Surface treatment System based on matrix, indices
and structure is not reported in literature. Graph theory application for system modeling of the
whole surface treatment System as in totality, based on the structure is found missing in the
literature. It is reported in the literature that the systems model have very high capability to
integrate the different components of the system [17, 18]. Thus, there is a need to consider all
components of the surface treatment system together so that overall performance out put can be
found out.

The systems modeling, analysis, and evaluation of different electroplating surface treatment
processes is to be carried out with the use of Taguchi, Graph theory and other systems
approaches. The set up of electroplating is available in the workshop of BITS Pilani Goa
Campus, for conducting experiment.
The various phases of the research activity are as described below:
Phase I: There would be an extensive literature survey in areas like total surface treatment
system and subsystem, research work done, concept and terminology to achieve proposed
objective. The existing methodology for selecting different parameters for electroplating process,
its development and evaluation will be studied. Literature survey will be carried out in the
relevant area such as process, equipment, effect on environment and system tools and techniques
used. Attributes of electroplating surface treatment processes and different subsystems are to be
identified for incorporating in proposed model.
Phase II: Mathematical model will be developed for the total electroplating system connecting
its sub systems and all possible interconnections. The research work would also focus on
formulation of a mathematical model by incorporating its various sub system and their attributes
using system approach.
Phase III: Reliability, modeling and validity analysis of electroplating surface treatment system
would be carried out through appropriate techniques like fuzzy MADM, system approach etc.
Cause and effect analysis of the system will be carried out to improve performance output.
Hierarchical representation of various sub systems and different attributes up to component level
for analysis, synthesis and evaluation of total surface treatment system will be developed.
Phase IV: Development of methodology for coding, evaluation, comparison and selection of
optimum electroplating system for different applications is proposed to carry out. This research
study will be helpful for consumer, supplier, manufacturer and designer at conceptual stage for
designing and selecting optimum electroplating surface treatment system. Design for safety and
process of effluent treatment will be carried out, so that the system must conform to the standard
of ISO 14000. Based on cause and effect and SWOT analysis, failure mode effect criticality
analysis of the system will be carried out to develop improved system model.
Phase V: Sensitivity analysis will be carried out to identify critical parameters for process and
system improvement. The parameters such as pH value of solution, area of anode exposed to

solution, distance between work piece and plating material etc. will be taken for experimentation.
The effect of parameters such as pH value of solution, area of anode exposed to solution,
distance between work piece and plating material, etc. will be studied during experimentation.
These parameters will be varied at different levels and experiment based on Taguchi orthogonal
array approach will be conducted in electroplating section of work shop of BITS Pilani Goa
campus. This in turn optimizes the influencing parameters, such that, the objectives like coating
thickness, deposition rate etc., will be less sensitive to uncontrolled factor (like environment
temperature, human error etc.). It is proposed to get the comments on the developed
methodology for its practicability, effectiveness and efficiency from the near by electroplating
Phase VI: In this last phase, suitable conclusions and/or recommendations based on above
analysis will be suggested with thesis writing and the final thesis would be submitted. It is
expected this research study will be useful to designer of electroplating surface treatment
product, processes and equipment.

Work Plan:
The activities scheduled and the time by which these are to be achieved are indicated in fig. 2

                                                                          Phase 6
     C                                                     Phase 5
     I                                                Phase 4
     I                                    Phase 3
     Y                       Phase 2

                                       Phase 1

          0           6           12             18        24           30            36

                                  DURATION IN MONTHS
                            Figure 2

  1. Dowey S. J., Matthews A. Taguchi and TQM: Quality issues for surface engineered
     applications. Surface and Coatings Technology. 1998, 110:86-93.

  2. Chan Lai-Kow, Wu Ming-Lu. Quality function deployment: A literature review.
     European Journal of Operational Research. 2002, 143:463-497.

  3. Flott L.W. Quality control: Professionalism in a changing world. Metal Finishing. 1998,

  4. Flott L.W. Quality Control: Six-Sigma Controversy. Metal Finishing. 2000, 98:43-48.

  5. Chan Felix T.S. Application of a hybrid case-based reasoning approach in electroplating
     industry. Expert Systems with Applications. 2005, 29:121-130.

  6. J R House. What's in or on a Surface? Surface engineering consultant Ltd. [serial on
     internet]. 2002-2003; [about 2 p.].

     Available from:

  7. Chan C.W., Huang Guo H. Artificial intelligence for management and control of
     pollution minimization and mitigation processes. Engineering Applications of Artificial
     Intelligence. 2003, 16:75-90.

  8. Lin J. L., Lin C. L. The use of grey-fuzzy logic for the optimization of the manufacturing
     process. Journal of Materials Processing Technology. 2005, 160:9-14.

  9. Flott L.W. Taguchi for fun and profit. Metal Finishing. 2002, 100:58-63.

  10. Jiang Ming, Komanduri R. Application of Taguchi method for optimization of finishing
     conditions in magnetic float polishing (MFP). Wear. 1997, 213:59-71.

  11. Mascio R. Di, Barton G. W. The economic assessment of process control quality using a
     Taguchi-based method. Journal of Process Control. 2001, 11:81-88.

12. Gaitonde V. N., Achyutha B. T., Siddeswarappa B. Burr size minimization in drilling
   using Taguchi technique. Indian Journal of Engineering & Material Sciences. 2005,

13. Nian C. Y., Yang W. H., Tarng Y. S. Optimization of turning operations with multiple
   performance characteristics. Technology. 1999, 15:90-96.

14. Singh H., Kumar P. Optimizing cutting force for turned parts by Taguchi’s parameter
   design approach. Indian Journal of Engineering & Material Sciences. 2005, 12:97-103.

15. Mohan M., Gandhi O.P., Agrawal V.P. Systems modeling of a coal-based steam power
   plant. Proceeding of Institution of Mechanical Engineers. UK. Part A: Power and Energy.
   2003, 217:259-277.

16. Mohan M., Gandhi O.P., Agrawal V.P. Maintenance strategy for a coal-based steam
   power plant equipment: a graph theoretic approach. Proceeding of Institution of
   Mechanical Engineers. UK. Part A: Power and Energy. 2004, 218:619-636.

17. Deo N. Graph Theory with applications to Engineering and Computer Science. Prentice-
   Hall of India Private Limited: New Delhi; 2004.

18. Gandhi O.P., Agrawal V.P. FMEA- A digraph and matrix approach. Journal of
   Reliability Engineering and System Safety. 1992, 35:147-158.

19. Gandhi O.P., Agrawal V.P., Shishodia K.S. Reliability Analysis and Evaluation of
   Systems. Journal of Reliability Engineering and System Safety. 1991, 32:283-305.

20. Gandhi O.P., Agrawal V.P. Failure Cause Analysis – A Structural Approach. Journal of
   Pressure Vessel Technology. 1996, 118:434-440.

21. Gandhi O.P., Agrawal V.P. A Digraph Approach to System Wear Evaluation and
   Analysis. Journal of Tribology, Transactions of the ASME. 1993, 93-Trib-54:1-5.

22. Khan I.A., Mubeen A., Agrawal V.P. SWOT Function Deployment to Maintenance
   Strategies. Journal of the Institute of Engineers (India). 2003, 83:41-45.

   23. Grover S., Agrawal V.P., Khan I.A. A digraph approach to TQM evaluation of an
       industry. International Journal of Production Research. 2004, 42:4031-4053.

   24. Venkatasamy R., Agrawal V.P. System and structural analysis of an automobile vehicle-
       a graph theoretic approach. International of Vehicle Design. 1995, 16:477-505.

   25. Venkatasamy R., Agrawal V.P. Computer Aided Evaluation and Selection of Automobile
       Vehicle by Fuzzy MADM Method. Mobility and vehicle Mechanics- KRAGUJEVAC.
       1995, 21:13-30.

   26. Venkatasamy R., Agrawal V.P. Computer Aided Evaluation and Selection of Plastic and
       Composite Material for Automobile Vehicle Component. Proceedings of 9 th World
       Congress on Theory of Machine and Mechanism, Milan, Italy: 1995, Aug. 30– Sept.

Brief Biodata of the Supervisor

Name of the candidate                              *************

Name of the supervisor                             Dr.*****************

ID of candidate                                    200*PHXF***

Designation and Address

Experience (years)

Number of Publications

No. of Ph.D. student supervised

                                   SAMPLE PROPOSAL 2
                       Outline of the Proposed Topic of Research

Name of Candidate                          : ***********
ID No.                                     : ***********
Place of Research Work and Organisation    : Institute of ********, New Delhi
Proposed Supervisor’s Details
Name                                       : ***************
Qualification                              : M.Sc., Ph.D
Designation                                : ***************
Organization                               : Institute of *********, New Delhi

Proposed Topic of Research
Role of Antigen Presenting Cells (APCs) and Toll like Receptors in Providing a Protective
Immune Response during Chlamydia trachomatis Infection

Objective of the Proposed Research
   1. In vitro study of processing and presentation of chlamydial antigens by Dendritic cells
         (DC’s) and monocytes/macrophages to CD8 + and CD4+ T lymphocytes.
   2. Differential regulation of cytokine production by CD8 + and CD4 + T lymphocytes.
   3. Role of Toll like receptors 2 and 4 in recognition of Chlamydial antigens and regulation
         of cytokine production.
   4. Regulation of nitric oxide production by chlamydial antigens.

Background of the Proposed Research
Worldwide, an estimated 90 million sexually transmitted Chlamydia trachomatis infections
occur each year. Sexually transmitted C. trachomatis infection is an important public health
concern because of its adverse effects on reproduction [1]. In India alone a high chlamydial
prevalence rate (28%) was found in symptomatic patients [2]. In women, infection with C.

trachomatis causes pelvic inflammatory disease (PID) and has long term consequences – such as
infertility, ectopic pregnancy and chronic pelvic pain- that are secondary to scarring of the
fallopian tubes (caused by salpingitis) and ovaries. In addition, infection with C. trachomatis
felicitates the transmission of HIV [3] and might be a co factor in human papilloma virus (HPV)-
induced cervical neoplasia [4-5]. The pathological mechanism by which C. trachomatis induces
scarring is not well understood. In all cases the pathology seems to be related to a chronic
inflammation caused by a persistent chlamydial infection or by repeated infections with the
After initial infection of the host with C. trachomatis, Dendritic cells (DC) are the first
professional antigen presenting cells (APC’s) encountering the bacteria. DC’s are present in the
epithelium of cervix and vagina [6] and they prime the T cells to modulate the type of T -cell
responses (Th1/Th2), contributing to the inflammatory response which largely depends on the
up-regulation of co-stimulatory and adhesion molecules and on secretion of inflammatory
cytokines [7-9]. Protection against chlamydial infection has been shown to be primarily mediated
by IFN- producing T-cells [10,11] and it has been shown that DC can process and present
chlamydial antigens to T cells [12-14]. Little is known, however, about the interaction between
human DC and Chlamydia species.
Chlamydia trachomatis is an obligate intracellular gram –ve bacterium which causes a wide
spectrum of human diseases. C. trachomatis infection of genital tract is now recognized as one of
the most common major cause of sexually transmitted diseases in developed countries [1].
Without treatment the chlamydial genital infections can cause pelvic inflammatory disease (PID)
and its sequelae of ectopic pregnancy and infertility.
The chlamydial developmental cycle involves a metabolically inactive non replicating infectious
form called elementary body (EB), that, after entry into the host cells differentiate into
metabolically active Reticulate Body (RB). The organism infects the epithelial cells often
inducing an acute inflammatory response, caused by persistent chlamydial infections, or by
repeated infections with the bacterium, however, protective immunity is limited.
C. trachomatis infection remains sub clinical in a high proportion of infected individuals (70-
90% of women and 30-50% of men). Clinical symptoms if present include dysuria, abnormal
vaginal discharge and lower abdominal pain. Infection ascends the endometrial epithelium to the

fallopian tubes, where C. trachomatis can establish persistent infection and can cause PID.
Overall, 11% of women with PID develop tubal factor infertility and 9% develop ectopic
Both humoral and cellular responses can be readily detected in patients suffering from C.
trachomatis infection. Since infection is intracellular, neutralizing antibodies have little
relevance in resolving infection [10]. An early epidemiological observation suggested an inverse
correlation between the amount of IgA in cervical secretions and the amount of C. trachomatis
recovered from the cervix of infected women [15]. In vitro, antibodies specific for C.
trachomatis can neutralize infection in tissue culture [16], however, in humans high titers of C.
trachomatis specific antibodies do not correlate with resolution of infection and , in fact, more
strongly correlated with increased severity of sequelae of infection, such as tubal infertility [17].
In contrast, there is good evidence that T-cell mediated immune (CMI) responses play a major
role in clearance and resolution of chlamydial infections and T-cell responses are critical in host
resistance to C. trachomatis. Transfer of T lymphocytes into naïve mice have been shown to
protect the mice against C. trachomatis infection [18].
After infection of host with C. trachomatis, dendritic cells (DC) and macrophages/monocytes are
the main antigen presenting cells (APC’s) encountering the bacteria. DC’s are key players in
immunity that dictate the type of immune response generated to a particular antigen. DC’s are
professional antigen presenting cells that have extraordinary capacity to stimulate naive T-cell
and initiate primary immune response. Immature DC present in the epithelium of cervix and
vagina capture microbial antigens, process them and present them to T lymphocytes. Mature
DC’s are highly effective at presenting antigen and priming protective adaptive immune
TLR’s comprise a family of cell surface receptors that recognize pathogen associated molecular
patterns (PAMP’s), including lipopolysaccharide (LPS) and hypomethylated CpG- rich DNA as
well as double stranded and single stranded RNA. Toll like receptors detect microbial infection
and have an essential role in the induction innate and adaptive immune responses [19]. Of the 10
cloned mammalian TLRs, TLR2 and TLR4 are the best characterized with respect to innate
responses to bacteria. TLR4, in association with accessory molecules MD-2 and CD 14, is the
signal transduction receptor for gram- negative bacterial lipopolysaccharide and heat shock
proteins. A broader range of microbial products activate immune responses through engagement

of TLR2, including peptidoglycan from gram-positive bacteria, bacterial lipopeptides, and
zymosan. A recent hypothesis states that differential expression and engagement of TLR family
members at the surface of dendritic cells and macrophages influences the type of immune
response that is induced by a microbial pathogen. Infection with Chlamydia muridarum has been
shown to stimulate DC’s to produce IL-12 (TH1 type response) [20]. It is not confirmed which
particular TLR’s expressed by dendritic cells are engaged by Chlamydia spp., TLR2 might have
an important role in the activation of DC’s by C. pneumoniae [21]. Furthermore, signaling
through TLR2 , but not TLR4, is associated with increase fallopian tube pathology in C.
muridarum infected mice [22], indicating that engagement of TLR2 is a potential common
pathway in both the immunity and immunopathology induced by Chlamydia spp. Given the high
level of expression of TLR’s by DC’s and there ability to polarize immune responses, the
identification of the role of DC’s in Chlamydia specific immune responses is crucial for
understanding the type of immune response that is elicited and therefore also for designing a
vaccine against infection with Chlamydia trachomatis [23].
Studies in the animal models have clearly established that T cells have a crucial role in the
resolution of infection with Chlamydia spp [24]. Nude mice cannot control infection and
adoptive transfer of Chlamydia specific CD4+ or CD8+ cells allow these mice to successfully
control infections. Specifically protection seems to be mediated by CD4 + T cells that produce
IFN- [25]. The role and effector mechanisms of Chlamydia positive CD8 + T cells are less clear.
MHC class I peptide presentation to CD8 + T cells is not essential for clearance of infection with
Chlamydia spp. In some situation CD8 + T cells might be important in elimination of cells
infected with Chlamydia spp. [26]. Also adoptive transfer of CD8 + T cell lines specific for
serovar L2 of C. trachomatis protected mice against infection with C. trachomatis through a
mechanism involving the production of IFN- [27]. Thus to establish the real role played by
CD8+ T cells in C. trachomatis infection further studies are required.
Stimulated T cells produce a variety of cytokines for clearance of Chlamydial infection including
IFN- and IL-12. Most of the cytokines secreted by T cells and macrophages are T helper1 (T H1)
cytokines, which have a role in polarizing the immune response to Chlamydia spp towards a
protective T H1 type response. By contrast, cytokines such as TNF, IL-1 and IL-10 might be
involved in the pathology associated with infection with Chlamydia spp. IFN-, the main T H1
type cytokine is essential for the clearance of Chlamydial infections from genital tract. It controls

the in vitro growth of C. trachomatis through inducing production of enzyme indoleamine-2,3-
dioxygenase (IDO) [28]. Activation of IDO by IFN- leads to degradation of tryptophan and lack
of this essential amino acid causes the death of C. trachomatis through tryptophan starvation
[28]. Additional immune effector mechanisms induced by IFN- include induction of nitric oxide
production, which inhibits growth of C. muridarum [29] and the promotion of T H1 type
protective immune response, which downregulate non protective T H2 type responses, thereby,
promoting persistent infection [30]. Persistent infection might induce the secretion of
proinflammatory cytokines, leading to chronic inflammatory cellular response and tissue damage
[31, 32]. Overall, these data show that Chlamydia specific CD4+ TH1 cells and to a more limited
extent CD8+ T cells are required to control C. muridarum infection in genital tract of mice [33].
Observations from humans infected with C. trachomatis indicate that similar immune effector
mechanisms occur in humans [33].
Gap in Existing research
More than two-third of the Chlamydial infections cases occur in the developing countries, where
diagnostic and treatment services are almost absent. An estimated 15 million new cases are
occurring in Africa and 45 million new cases in Southern Asia every year [34]. Because of its
effect on reproduction, programmes to control C. trachomatis have been implemented in many
developed countries but many regions are now showing an increase in the number of infected
individuals [35]. Since current programmes for the control of C. trachomatis are not affordable
for much of the developing countries, vaccine development have been identified as an essential
to controlling infection with C. trachomatis. Mouse models of chlamydial infections with C.
muridarum have provided information on the immune mechanisms of clearance of infection and
resistance to reinfection, but there are several important differences between C. muridarum and
C. trachomatis that might effect the immunobiology of infection. Firstly, C. trachomatis
infection in humans is much more prolonged than C. muridarum infection in mice [36].
Secondly, immune-evasion mechanisms also differ. These differences limit the direct
extrapolation of findings from C. muridarum infection to C. trachomatis infection. Thus, a better
definition of human immune response correlates with C. trachomatis protective immunity and
disease pathogenesis needs to remain an important research priority if we are to develop a
vaccine against C. trachomatis infection that has protective and not deleterious effects.

For carrying out the proposed research work facilities such as tissue culture room equipped with
bio safety hood and incubator, thermocyclers for Polymerase Chain reaction, cell sorter, flow
cytometer, confocal microscope and computers for analysis and storage of data are required
which are available at the Institute of Pathology.
Phase 1:
This phase will comprise of Literature survey.
Phase 2:
(A) Enrollment of patients:
Symptomatic female patients attending the Gynecology Out Patient Department of Safdarjung
Hospital, New Delhi, and having complaints of cervical/ vaginal discharge, abdominal pain,
dysuria or infertility will be enrolled. Cervical lavage samples and peripheral blood will be
collected from these women. Diagnosis for Chlamydia trachomatis and other STD pathogen will
be done using standard methodology.
(B) Flow Cytometry:
Quantification of different T-cell subsets, monocytes, dendritic cells and expression of TLR’s on
the surface of dendritic cells and monocytes will be performed by standard flow cytometry
Phase 3: Culture of Monocyte derived Dendritic Cells (MDDC)
Peripheral blood mononuclear cells (PBMC’s) will be purified on Ficoll gradients and will be
washed three times with RPMI 1640. They will be then stained with CD14 microbeads and will
be separated using a cell sorter. These CD14 positive cells will be inoculated (1 -2 x 106/ml) into
each well of a six well plate and will be cultured in RPMI 1640 supplemented with 10% heat
inactivated Fetal Calf Serum, 2mM L-glutamine, 25mM HEPES, 0.02M 2-mercaptoethanol,
10g/ml Gentamycin and 2g/ml Amphotericin B at 37 0 c in a 5% CO2 incubator, in the
presence of 50 ng/ml Granulocyte Macrophage Colony Stimulating factor and 20 ng/ml IL-4 for
6-7 days. After 6-7 days immature MDDC’s will be washed and analyzed for CD14 and CD1a
expression (marker for immature DC’s)
Phase 4: Coculture of Dendritic cells with CD8 and CD4 T lymphocytes
Immature Dendritic cells and Monocytes will be infected with live C. trachomatis EB’s and
cultured in RPMI 1640 for 4 days at 37 0C. Autologous CD4 + and CD8+ T cells will be separated

using a cell sorter and will be cocultured with the EB pulsed dendritic cells and monocytes for
further 4 days in the presence of recombinant IL-2 for generation of T-cell clones. The culture
supernatants will be then analyzed for production of various cytokines.
Phase 5:
(A) Blocking of Toll like receptors on dendritic cells and monocytes
Toll like receptors 2 and 4 present on the surface of Dendritic cells and monocytes will be
blocked with the help of blocking peptides before pulsing them with chlamydial EB’s and will
then be cocultured with CD8 + and CD4+ T cells. The culture supernatants will be again analyzed
for production of various cytokines. Changes in the gene expression patterns of Toll like
receptors, upon infection with Chlamydial EB’s will be done with RT-PCR (reverse transcriptase
Polymerase chain reaction).
(B) Regulation of nitric oxide production by chlamydial antigens
Nitric oxide production by antigen presenting cells and T cells will be analyzed in the culture
supernatants and its regulation by IFN- and Toll like receptors will be studied.
Phase 6: Conclusion and Thesis writing
Results will be concluded with the help of the data, which we obtained throughout our research
work and will be compiled in thesis.
Work Plan :

                                                                              PHASE 6

                                                                    PHASE 5
                                                               PHASE 4
                                                   PHASE 3
                Y                         PHASE 2

                                         PHASE 1

                     0        6     12       18           24      30     36       42    48
                                           Duration in Months

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Description: Introduction lu vaccine