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. 2019 Dec;98(49):e18073.
doi: 10.1097/MD.0000000000018073.

Polymorphism of MPT64 and PstS1 in Mycobacterium tuberculosis is not likely to affect relative immune reaction in human

Tongyang Xiao  1   2   3 , Yi Jiang  1   2 , Guilian Li  1   2 , Hui Pang  4 , Lili Zhao  1   2 , Xiuqin Zhao  1   2 , Kanglin Wan  1   2
Affiliations

Polymorphism of MPT64 and PstS1 in Mycobacterium tuberculosis is not likely to affect relative immune reaction in human

Tongyang Xiao et al. Medicine (Baltimore). 2019 Dec.

Abstract

Background: MPT64 and PstS1 are the earliest known immune-dominant antigens of Mycobacterium tuberculosis and have been commonly used as candidates in the diagnosis of tuberculosis.

Methods: We constructed recombinant plasmids pET-32a-Rv0934 and pET-32a-Rv1980c to express both wild and mutant forms of MPT64 and PstS1 and purified them. From November 9 to December 9, 2016, and November 9 to December 10, 2017, 96 patients with tuberculosis, 53 patients without tuberculosis, and 96 healthy volunteers were enrolled in this study. We used the purified proteins as antigens to perform T-spot and enzyme-linked immunosorbent assay (ELISA) for samples obtained from healthy volunteers and tuberculosis patients.

Results: Regarding T-spot, the area under the curve (AUC) values for MPT64-wild protein (MPT64-H37Rv) and MPT64-mutant protein (MPT64-FJ05395) were 0.723 and 0.750, respectively. The AUC values for PstS1-H37Rv, PstS1-FJ05132, and PstS1-JL06035 were 0.817, 0.796, and 0.745, respectively. With regard to ELISA, the AUC values for MPT64-H37Rv and MPT64-FJ05395 were 0.525 and 0.528, respectively, while those for PstS1-H37Rv, PstS1-FJ05132, PstS1-JL06035 were 0.588, 0.509, and 0.560, respectively. There was no difference between wild and mutant proteins when we used them as antigens to perform T-spot and ELISA assays.

Conclusion: MPT64 and PstS1 are likely candidate diagnostic antigens for M tuberculosis T-spot test, at least in combination with other proteins. Polymorphisms of MPT64 and PstS1 had little effect on cell-mediated and humoral immunity in the host.

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Conflict of interest statement

The authors have no conflicts of interest to disclose.

Figures

Figure 1
Figure 1
SDS-PAGE of purified recombinant wild and mutant proteins expression. Lanes: 1, Standard protein marker; 2, induced pET-32a-PstS1-H37Rv; 3, induced pET-32a-PstS1-FJ05132; 4, induced pET-32a-PstS1-JL06035; 5, induced pET-32a-MPT64-H37Rv; 6, induced pET-32a-MPT64-FJ05395. SDS-PAGE = sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
Figure 2
Figure 2
ROC curve comparison of wild and mutant MPT64 proteins for T-spot (wild form: PstS1-H37Rv; mutant form: PstS1-FJ05132 and PstS1-FJ06035). ROC = receiver operating characteristic.
Figure 3
Figure 3
ROC curve comparison of wild and mutant PstS1 proteins for T-spot (wild form: PstS1-H37Rv; mutant form: PstS1-FJ05132 and PstS1-FJ06035). ROC = receiver operating characteristic.
Figure 4
Figure 4
ROC curve comparison of wild and mutant MPT64 proteins for ELISA (wild form: MPT64-H37Rv; mutant form: MPT64-FJ05395). ELISA = enzyme-linked immunosorbent assay; ROC = receiver operating characteristic.
Figure 5
Figure 5
ROC curve comparison of wild and mutant PstS1 proteins for ELISA (wild form: MPT64-H37Rv; mutant form: MPT64-FJ05395). ELISA = enzyme-linked immunosorbent assay; ROC = receiver operating characteristic.
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