IgA and IgG against Mycobacterium tuberculosis Rv2031 discriminate between pulmonary tuberculosis patients, Mycobacterium tuberculosis-infected and non-infected individuals

Abstract

As part of a major project to investigate protective and diagnostic immune markers against tuberculosis (TB), we measured antibody isotype responses to Mycobacterium tuberculosis (Mtb) antigens (LAM, Rv2031, and HBHA) in cohorts of 149 pulmonary tuberculosis patients (PTBP), 148 household contacts (HHCs), and 68 community controls (CCs) in an endemic setting. ELISA was used to measure levels of IgA, IgG, and IgM from sera of cohorts at baseline, and at 6 and 12 months from entry. The results show that there were significant differences in IgA, IgG, and IgM responses to the different antigens and in the three cohorts. At baseline, the level of IgM against RV2031 and LAM did not vary between cohorts, but the levels of IgA and IgG against Rv2031 were significantly higher in PTB patients than HHCs and CCs, followed by HHCs, and the lowest in CCs. In patients, there was a significant variation in antibody responses before and after chemotherapy. The levels of IgA and IgG against HBHA, and IgA against Rv2031 decreased significantly and remained low, while IgA and IgG against LAM increased significantly and remained high following chemotherapy. However, the levels of IgM against Rv2031 and LAM increased at 6 months but decreased again at 12 months. IgM against HBHA did not show any significant variation before and after chemotherapy. Similarly, there were also significant variations in antibody responses in HHCs over time. Our results show that there are significant variations in IgA, IgG and IgM responses to the different antigens and in the three cohorts, implying that not all antibody isotype responses are markers of clinical TB. In addition, the current and previous studies consistently show that IgA and IgG against Rv2031 discriminate between clinical disease, Mtb-infected and non-infected individuals.

Fig 1

(a-c). IgG responses to LAM at baseline in the three cohorts (Fig a), 6 and 12 months in patients (Fig b) and HHCs (Fig c). Results are individual responses expressed as OD values. The median for each group is shown as horizontal bar. Kruskal-Wallis and Friedman test with Dunn`s multiple comparisons were used to compare antibody responses among groups and over time, respectively. *p<0.05; ****p<0.0001.

Fig 2

(a-c). IgA responses to LAM at baseline in the three cohorts (Fig a), 6 and 12 months in patients (Fig b) and HHCs (Fig c). Results are individual responses expressed as OD values. The median for each group is shown as horizontal bar. Kruskal-Wallis and Friedman test with Dunn`s multiple comparisons were used to compare antibody responses among groups and over time, respectively. **p<01*p; ***p<0.001; ****p<0.0001.

Fig 3

(a-c). IgM responses to LAM at baseline in the three cohorts (Fig a), 6 and 12 months in patients (Fig b) and HHCs (Fig c). Results are individual responses expressed as OD values. The median for each group is shown as horizontal bar. Kruskal-Wallis and Friedman test with Dunn`s multiple comparisons were used to compare antibody responses among groups and over time, respectively. ***p<0.001; ****p<0.0001.

Fig 4

(a-c). IgG responses Rv2031 at baseline (Fig a), 6 and 12 months in patients (Fig b) and HHCs (Fig c). Results are individual responses expressed as OD values. The median for each group is shown as horizontal bar. Kruskal-Wallis and Friedman test with Dunn`s multiple comparisons were used to compare antibody responses among groups and over time, respectively.*p0.05; **p<0.01; ***p<0.001; ****p<0.0001.

Fig 5

(a-c). IgA responses to Rv2031 at baseline (Fig a), 6 and 12 months in patients (Fig b) and HHCs (Fig c). Results are individual responses expressed as OD values. The median for each group is shown as horizontal bar. Kruskal-Wallis and Friedman test with Dunn`s multiple comparisons were used to compare antibody responses among groups and over time, respectively. *p<0.05; ****p<0.0001.

Fig 6

(a-c). IgM responses to Rv2031 at baseline (Fig a), 6 and 12 months in patients (Fig b) and HHCs (Fig c). Results are individual responses expressed as OD values. The median for each group is shown as horizontal bar. Kruskal-Wallis and Friedman test with Dunn`s multiple comparisons were used to compare antibody responses among groups and over time, respectively. ****p<0.0001.

Fig 7

(a-c). IgG responses to HBHA at baseline (Fig a), 6 and 12 months in patients (Fig b) and HHCs (Fig c). Results are individual responses expressed as OD values. The median for each group is shown as horizontal bar. Kruskal-Wallis and Friedman test with Dunn`s multiple comparisons were used to compare antibody responses among groups and over time, respectively. ****p<0.0001.

Fig 8

(a-c). IgM responses to HBHA at baseline (Fig a), 6 and 12 months in patients (Fig b) and HHCs (Fig c). Results are individual responses expressed as OD values. The median for each group is shown as horizontal bar. Kruskal-Wallis and Friedman test with Dunn`s multiple comparisons were used to compare antibody responses among groups and over time, respectively. ****p<0.0001.