Biomedical Chemistry: Research and Methods 2019, 2(4), e00113

Transcriptional Analysis of HeLa Cells - Producers of the Recombinant Peptidoglycan Recognition Protein PGLYRP1 at Different Stages of the Chlamydia Trachomatis Infection Development

P.A. Bobrovsky*, A.K. Larin, N.F. Polina, V.N. Lazarev

Federal Research and Clinical Center of Physical-Chemical Medicine,
1a Malaya Pirogovskaya str., Moscow, 119435 Russia; *e-mail: pbobrovskiy@gmail.com

Keywords:Chlamydia trachomatis; PGLYRP; recombinant proteins; transcriptomics

DOI:10.18097/BMCRM00113

The whole version of this paper is available in Russian.

Human peptidoglycan recognition proteins (PGLYRPs) are the components of innate immunity that exhibit antibacterial activity. In this study a cell line secreting recombinant PGLYRP1 into a culture medium was obtained. Transcriptional profiling of cell lines expressing PGLYRP1 was performed at different stages of C. trachomatis infection. Differential gene expression was studied using the whole transcriptome profiling method on the HumanHT-12 v4 Expression BeadChip microchip using the Illumina Direct Hybridization Whole-Gene Expression Assay protocol. Sample clustering followed by bioinformatics analysis revealed about 100 differentially expressed genes in response to infection with C. trachomatis. PGLYRP1- expressing cells infected with C. trachomatis had a similar transcriptional profile as non-infected cells.
Figure 1. Western blot analysis of culture medium from HeLa-PGLYRP1 and HeLa-PGLYRP1m cell lines. The membrane was incubated with anti-6His monoclonal antibodies. Lanes 1, 2, 3 – 50, 25, 10 ng of purified recombinant PGLYRP1, lanes 4, 5, 6 – three clones of HeLa-PGLYRP1,
7 - HeLa-PGLYRP1m, the molecular mass markers are shown on the right (kilodaltons).
Figure 2. Sample cluster analysis. All data from the two biochips were clustered by the similarity of fluorescence values after sorting the data by the significance level of the detected fluorescence signal (p≤0.05).
Figure 3. The heat map of the correlation analysis of the studied samples. On the left is the sample cluster analysis. The correlation was calculated for all 24 samples from two biochips after sorting the data by the significance level of the detected fluorescence signal (p≤0.05).
Figure 4. The result of differentially expressed genes in GO terms annotation. The data are presented for one group of gene ontology: the cellular component. Part A corresponds to data for 1 hour after infection, part B corresponds to data for 72 hours after infection. FDR – significance level.
Figure 5. The Venn diagram for the search for common differentially expressed HeLa-PGLYRP1m cell genes at 1, 24 and 72 hours after infection with C.trachomatis. The diagram is based on differentially expressed genes that differ from control by 3 or more times.

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Table 1. Scheme of an experiment of transcriptome profiling of cell lines expressing PGLYRP1 after infection with C. trachomatis at different stages of infection.

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Table 2. List of common differentially expressed genes in HeLa-PGLYRP1m cell line 1, 24 and 72 hours after infection with C. trachomatis.

SUPPLEMENTARY

Supplementary materials are available at http://dx.doi.org/10.18097/BMCRM00113

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