Immunological and proteomic analysis of preparative isoelectric focusing separated culture filtrate antigens of Mycobacterium tuberculosis

Deenadayalan, Anbarasu and Sundaramurthi, J C and Raja, Alamelu (2010) Immunological and proteomic analysis of preparative isoelectric focusing separated culture filtrate antigens of Mycobacterium tuberculosis. Experimental and Molecular Pathology, 88 (1). pp. 156-162. ISSN Print: 0014-4800

Archive (PMC) 201010.pdf - Published Version Restricted to Registered users only Download (411kB) | Request a copy

Abstract

Isolation of the secreted proteins and studying the immune response they induce is an essential prerequisite for understanding the pathogenesis of M. tuberculosis. In this study, preparative liquid-phase isoelectric focusing was used for the separation of culture filtrate protein (CFP) of M. tuberculosis. This procedure resolved culture filtrate proteins into 20 fractions with a pI range of 2.59 to 12.9. These 20 fractions were subjected to immunological analysis in healthy laboratory volunteers from our endemic area. Eleven fractions (Fractions 5, 6, 7, 8, 9, 10, 11, 13, 15, 16, and 19) showed increased interferon gamma (IFN-γ) secretion and 5 fractions induced increased proliferative response, when compared to unfractionated CFP. In the 11 fractions which showed increased IFN-γ secretion, mass spectrometric analysis identified 19 different proteins. Apart from the already reported immunodominant antigens like FbpB, CFP-10 and ESAT-6, two new T cell antigens (AcpM and PpiA) were also identified in the immunologically active fractions. Immunoinformatic analysis showed that PpiA was predicted to bind more number of class I and class II HLA alleles compared with the immunodominant ESAT-6 and CFP-10. Population coverage calculations also showed that PpiA protein (85%) had a higher population coverage compared with ESAT-6 (79%) and CFP-10 (77%). This result shows that the PpiA protein has a potential to be a novel T cell antigen.

Affiliation:	National Institute for Research in Tuberculosis
Item Type:	Article
Uncontrolled Keywords:	Culture filtrate proteins Isoelectric focusing Mass spectrometry Rotofor Tuberculosis
Subjects:	Tuberculosis > Laboratory Research > Immunological
Divisions:	Basic Science Research > Immunology
Depositing User:	Dr. Rathinasabapati R
Date Deposited:	17 Mar 2022 10:52
Last Modified:	17 Mar 2022 10:52
URI:	http://eprints.nirt.res.in/id/eprint/1026

Actions (login required)

View Item