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Differential expression of immunogenic proteins on virulent Mycobacterium tuberculosis clinical isolates.

Schierloh P, Klepp L, Vazquez C, Rocha RV, Blanco FC, Balboa L, López B, Ritacco V, Bigi F, Sasiain Mdel C - Biomed Res Int (2014)

Bottom Line: Recent studies have demonstrated that Mtb families found worldwide today differ in pathology, transmissibility, virulence, and development of immune response.We found that these proteins induced humoral immune responses in patients with drug-sensitive and drug-resistant tuberculosis with substantial cross-reactivity among the four proteins.Therefore, the present results describe new Mtb antigens with potential application as biomarkers of TB.

View Article: PubMed Central - PubMed

Affiliation: IMEX-CONICET, Academia Nacional de Medicina, Pacheco de Melo 3081, 1425 CABA, Argentina.

ABSTRACT
Molecular epidemiology has revealed that Mycobacterium tuberculosis (Mtb), formerly regarded as highly conserved species, displays a considerable degree of genetic variability that can influence the outcome of the disease as well as the innate and adaptive immune response. Recent studies have demonstrated that Mtb families found worldwide today differ in pathology, transmissibility, virulence, and development of immune response. By proteomic approaches seven proteins that were differentially expressed between a local clinical isolate from Latin-American-Mediterranean (LAM) and from Haarlem (H) lineages were identified. In order to analyze the immunogenic ability, recombinant Rv2241, Rv0009, Rv0407, and Rv2624c proteins were produced for testing specific antibody responses. We found that these proteins induced humoral immune responses in patients with drug-sensitive and drug-resistant tuberculosis with substantial cross-reactivity among the four proteins. Moreover, such reactivity was also correlated with anti-Mtb-cell surface IgM, but not with anti-ManLAM, anti-PPD, or anti-Mtb-surface IgG antibodies. Therefore, the present results describe new Mtb antigens with potential application as biomarkers of TB.

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Cross-reactivity analysis of humoral immune responses in drug-sensitive and MDR-TB patients' sera. (a) 2D scatter plots of combinatorial correlations among results against protein-specific and complex Ags in TB patients' serum samples (n = 37). Pearson correlation test was applied after verification of normal distribution of all data sets. “ns” indicates P > 0.05. Thresholds are delimited by dotted lines. (b) FACS analysis of cell surface binding-IgM and -IgG to LAM10406 in sera from TB patients (n = 37). Histogram plots of two TB and one No-TB patients are showed for comparison. Correlation between median fluoresce intensity (MFI) and protein-specific Abs (Pearson correlation test). “ns” indicates P > 0.05. Thresholds are delimited by dotted lines. (c) Antigens were classified as function of their pattern of seroreactivity in the 37 TB patients (hierarchical clustering based on the correlation). Matrix of 0 to 1 rescaled values is represented as heat-map.
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fig4: Cross-reactivity analysis of humoral immune responses in drug-sensitive and MDR-TB patients' sera. (a) 2D scatter plots of combinatorial correlations among results against protein-specific and complex Ags in TB patients' serum samples (n = 37). Pearson correlation test was applied after verification of normal distribution of all data sets. “ns” indicates P > 0.05. Thresholds are delimited by dotted lines. (b) FACS analysis of cell surface binding-IgM and -IgG to LAM10406 in sera from TB patients (n = 37). Histogram plots of two TB and one No-TB patients are showed for comparison. Correlation between median fluoresce intensity (MFI) and protein-specific Abs (Pearson correlation test). “ns” indicates P > 0.05. Thresholds are delimited by dotted lines. (c) Antigens were classified as function of their pattern of seroreactivity in the 37 TB patients (hierarchical clustering based on the correlation). Matrix of 0 to 1 rescaled values is represented as heat-map.

Mentions: One interesting observation from Table 2 is the abundance of MDR-TB patients' sera that react with more than one recombinant protein. In order to test if cross-reactivity is confined to recombinant proteins and if it is a specific feature of MDR-TB patients, we extended our analysis to other Mtb-Ags and to drug-sensitive TB patients. As can be observed in (Figure 4(a)), a strong correlation in Ab titers among each of the recombinant proteins was observed, but not with PPD or ManLAM Ags. Next, we tested if IgM and IgG Abs that bind Mtb-cell surface exposed structures were correlated with those directed to recombinant proteins. As it is shown in Figure 4(b), Ab titers against proteins correlated with anti-Mtb IgM but not with anti-Mtb-IgG. By multivariate based hierarchical classification, we concluded that Rv0407, Rv0009, Rv2624c, Rv2241, and Mtb-IgM Ags may be clustered together as function of their seroreactivity (Figure 4(c)). Analyses of antigen recognition by individual sera revealed that the pattern of antigens reacting with serum Abs varied greatly from patient to patient; thereby, no clear division among patients with MDR versus drug-sensitive as well as with bacillary loads was observed (Table 2).


Differential expression of immunogenic proteins on virulent Mycobacterium tuberculosis clinical isolates.

Schierloh P, Klepp L, Vazquez C, Rocha RV, Blanco FC, Balboa L, López B, Ritacco V, Bigi F, Sasiain Mdel C - Biomed Res Int (2014)

Cross-reactivity analysis of humoral immune responses in drug-sensitive and MDR-TB patients' sera. (a) 2D scatter plots of combinatorial correlations among results against protein-specific and complex Ags in TB patients' serum samples (n = 37). Pearson correlation test was applied after verification of normal distribution of all data sets. “ns” indicates P > 0.05. Thresholds are delimited by dotted lines. (b) FACS analysis of cell surface binding-IgM and -IgG to LAM10406 in sera from TB patients (n = 37). Histogram plots of two TB and one No-TB patients are showed for comparison. Correlation between median fluoresce intensity (MFI) and protein-specific Abs (Pearson correlation test). “ns” indicates P > 0.05. Thresholds are delimited by dotted lines. (c) Antigens were classified as function of their pattern of seroreactivity in the 37 TB patients (hierarchical clustering based on the correlation). Matrix of 0 to 1 rescaled values is represented as heat-map.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4109345&req=5

fig4: Cross-reactivity analysis of humoral immune responses in drug-sensitive and MDR-TB patients' sera. (a) 2D scatter plots of combinatorial correlations among results against protein-specific and complex Ags in TB patients' serum samples (n = 37). Pearson correlation test was applied after verification of normal distribution of all data sets. “ns” indicates P > 0.05. Thresholds are delimited by dotted lines. (b) FACS analysis of cell surface binding-IgM and -IgG to LAM10406 in sera from TB patients (n = 37). Histogram plots of two TB and one No-TB patients are showed for comparison. Correlation between median fluoresce intensity (MFI) and protein-specific Abs (Pearson correlation test). “ns” indicates P > 0.05. Thresholds are delimited by dotted lines. (c) Antigens were classified as function of their pattern of seroreactivity in the 37 TB patients (hierarchical clustering based on the correlation). Matrix of 0 to 1 rescaled values is represented as heat-map.
Mentions: One interesting observation from Table 2 is the abundance of MDR-TB patients' sera that react with more than one recombinant protein. In order to test if cross-reactivity is confined to recombinant proteins and if it is a specific feature of MDR-TB patients, we extended our analysis to other Mtb-Ags and to drug-sensitive TB patients. As can be observed in (Figure 4(a)), a strong correlation in Ab titers among each of the recombinant proteins was observed, but not with PPD or ManLAM Ags. Next, we tested if IgM and IgG Abs that bind Mtb-cell surface exposed structures were correlated with those directed to recombinant proteins. As it is shown in Figure 4(b), Ab titers against proteins correlated with anti-Mtb IgM but not with anti-Mtb-IgG. By multivariate based hierarchical classification, we concluded that Rv0407, Rv0009, Rv2624c, Rv2241, and Mtb-IgM Ags may be clustered together as function of their seroreactivity (Figure 4(c)). Analyses of antigen recognition by individual sera revealed that the pattern of antigens reacting with serum Abs varied greatly from patient to patient; thereby, no clear division among patients with MDR versus drug-sensitive as well as with bacillary loads was observed (Table 2).

Bottom Line: Recent studies have demonstrated that Mtb families found worldwide today differ in pathology, transmissibility, virulence, and development of immune response.We found that these proteins induced humoral immune responses in patients with drug-sensitive and drug-resistant tuberculosis with substantial cross-reactivity among the four proteins.Therefore, the present results describe new Mtb antigens with potential application as biomarkers of TB.

View Article: PubMed Central - PubMed

Affiliation: IMEX-CONICET, Academia Nacional de Medicina, Pacheco de Melo 3081, 1425 CABA, Argentina.

ABSTRACT
Molecular epidemiology has revealed that Mycobacterium tuberculosis (Mtb), formerly regarded as highly conserved species, displays a considerable degree of genetic variability that can influence the outcome of the disease as well as the innate and adaptive immune response. Recent studies have demonstrated that Mtb families found worldwide today differ in pathology, transmissibility, virulence, and development of immune response. By proteomic approaches seven proteins that were differentially expressed between a local clinical isolate from Latin-American-Mediterranean (LAM) and from Haarlem (H) lineages were identified. In order to analyze the immunogenic ability, recombinant Rv2241, Rv0009, Rv0407, and Rv2624c proteins were produced for testing specific antibody responses. We found that these proteins induced humoral immune responses in patients with drug-sensitive and drug-resistant tuberculosis with substantial cross-reactivity among the four proteins. Moreover, such reactivity was also correlated with anti-Mtb-cell surface IgM, but not with anti-ManLAM, anti-PPD, or anti-Mtb-surface IgG antibodies. Therefore, the present results describe new Mtb antigens with potential application as biomarkers of TB.

Show MeSH
Related in: MedlinePlus