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Summer Research Fellowship Programme of India's Science Academies

INTO THE ABODE OF IMMUNOLOGICAL TECHNIQUES

ANJITHA R.

Central University of Kerala, Kasaragod

PROF. AMITA AGGARWAL

Professor, Department of Clinical Immunology, Sanjay Gandhi Postgraguate Institute of Medical Sciences, Lucknow 226 014

Abstract

In this work, the various kinds of techniques applied in the field of immunology are being outlined. This work reveals from the simple and basic methods that we perform in our labs to the most recent and highly sophisticated techniques. Isolation of PBMC, DNA and RNA from the blood samples can be used as valuable samples to diagnose diseases and know the state of immunity in an individual. Here, flow cytometrical analysis of cells, DHR assay, Ki67 assay, RT-PCR, reverse transcription, ELISA, nephelometry etc. are performed for clinical diagnosis of certain conditions. In rheumatology, these are essential immunological techniques that serve for clinical purposes. In this, the project work that elucidates the effect of Rheumtoid Arthritis synovial fluid on the Th17/Treg ratio is also included. It tries to investigate the change that happens to the Th17/Treg ratio in a healthy blood sample on the addition of cell-free synovial fluid of Rheumatoid Arthritis patients.

Keywords: Immunology, Techniques, Rheumatoid Arthritis, Th17, Treg, Synovial fluid.

ABSTRACT

The synovial fluid of Rheumatoid Arthritis patients contains many proinflammatory cytokines like IL-1β, IL-23, IL-15, IFN-ϒ, TNF-α etc. The peripheral blood sample of RA patients was found to have disturbed Th17/Treg balance, due to the expansion of Th17 cells and Th17-related proinflammatory cytokines. Decreased frequency of Treg cells have also been observed in RA. Th17 causes an inflammatory response by release of proinflammatory cytokines, whereas Tregs have the role of immunotolerance, suppressing inflammatory activities. Th17 /Treg balance is very important for a healthy immunoregulation. A disruption in this pattern can lead to the development of autoimmunity. This work aims to investigate the effect of RA synovial fluid on the Th17/Treg ratio in healthy blood sample. This study infers to know whether the proinflammatory cytokines present in RA synovial fluid cause the polarization of T cells and leads to the expansion of Th17 cells in a healthy individual by assessing its markers in Flow Cytometer.

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