Identification and differential expression of serotransferrin and apolipoprotein A-I in the plasma of HIV-1 patients treated with first-line antiretroviral therapy

Abstract

Background As plasma proteins were involved in drug metabolism, the aim of the study was to identify and characterize the plasma proteins of the drug resistant and drug respondent groups of HIV-1 infected first line ART patients with > 6 years in therapy. Methods Plasma proteomics analysis of the four drug-resistant (treatment failure) and four drug respondent (treatment responder) groups were conducted. High abundant plasma proteins like albumin and globulin were depleted from plasma through Aurum serum mini kit (Bio-Rad, USA). Plasma proteins were resolved using two-dimensional gel electrophoresis on IPG strips, pH range of 3–10. Eight protein spots were selected in the gel based on the density of staining which was common in the drug resistant and drug respondent groups separately. The fold change of each spot was calculated using image-J. Each protein spot was identified using the matrix assisted laser desorption/ionization-time of flight/time of flight (MALDI-TOF/TOF) after tryptic digestion. Peptide peaks were identified through flex analysis version 3.3, and a search against a protein database using the internal mascot search engine. Gene ontology study was completed through STRING v.11 and Panther 15.0. Results Out of eight spots from 2D gel of drug respondent and drug resistant samples analyzed by MALDI TOF/TOF, two proteins were found to have significant score (i;e > 56) after Flex analysis. These two proteins were identified to be apolipoproteinA1 and Serotransferrin. The fold change expression of these two proteins were analyzed in drug resistant and drug respondent group. ApolipoproteinA1and serotransferrin were observed to be expressed 1.76 and 1.13-fold more respectively in drug respondent group compared to drug resistant group. The gene ontology analysis revealed the involvement of these two proteins in various important physiological processes. Conclusion Apolipoprotein A-I and serotransferrin were found to be expressed more in drug respondent group compared to drug resistant group.

Comparative evaluation of plasma protein purification and 2-D gel electrophoresis protocols for analysis of HIV-1 infected human plasma proteins

“…Even we used the broad range of isoelectric focusing gradient strip(IPG, pH 4-7) in this several experimental protocol, the suitable protocol is the outcome in the same IPG and also sensitive for mass spectrometry analysis such as MALDI-TOF-TOF and SWATH-MS. The excellent electrophoretic outcome was obtained during the identification and characterization of serotransferrin and apolipoprotein A1 in HIV-1 patients treated with first line ART (Barik et al, 2020 ) . High resolution two-dimensional gel electrophoresis of plasma protein is essential for identification of known and unknown components in the plasma(Anderson et al, 1977).…”

Identification and characterization of novel plasma proteins in drug resistant HIV/AIDS patients by SWATH-MS

“…The differential expression and molecular function of the Serotransferrin and Apolipoprotein A1 was identified in HIV-1 patients treated with first line ART 9 . Now, the fundamental role of these six statistically significant differentially expressed proteins such as complement C4-A, immunoglobulin heavy variable 1-2, carboxylic ester hydrolase, fibulin-1, immunoglobulin lambda constant 7, secreted phosphoprotein 24 and their association with several human diseases were discussed.…”

“…The genotyping was performed with samples with a viral load ≥ 1000 copies/ml at the National Institute for Research in Tuberculosis, Chennai, by using the WHO dried blood spot protocol 2010 8 . The details of the PCR primers and reaction conditions are reported earlier 9 .…”