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Drug resistant clinical isolates of Mycobacterium tuberculosis from different genotypes exhibit differential host responses in THP-1 cells.

Chakraborty P, Kulkarni S, Rajan R, Sainis K - PLoS ONE (2013)

Bottom Line: EAI-5 strain from ancient lineage 1, induced higher proinflammatory responses, higher apoptosis and moderate intracellular growth compared to other strains, in contrast, for Beijing strain of modern lineage 2, all three parameters were lowest among the clinical isolates.Thus, these profiles were specific to their respective lineages and/or genotypes and independent of their drug resistance status.Further, a positive correlation, among TNF-α, IL-1β, IL-6 and IL-12 induced in infected THP-1 cells was demonstrated.

View Article: PubMed Central - PubMed

Affiliation: Radiation Medicine Centre, Bio-Medical Group, Bhabha Atomic Research Centre, Mumbai, India.

ABSTRACT
Mycobacterium tuberculosis (MTB) persistently infects and survives within the host macrophages. Substantial genotypic variation exists among MTB strains which correlate with their interactions with the host. The present study was designed to establish a correlation, if any, between infection and induction of innate immune response by genetically diverse drug resistant MTB isolates from India. For this purpose, three clinical isolates from ancient and modern lineages, along with H37Ra and H37Rv were evaluated for intracellular growth, phagocytic index, induction of proinflammatory cytokines and apoptosis following infection in THP-1 cell line. A wide variation in the induction of cytokines was revealed subsequent to infection with different strains. EAI-5 strain from ancient lineage 1, induced higher proinflammatory responses, higher apoptosis and moderate intracellular growth compared to other strains, in contrast, for Beijing strain of modern lineage 2, all three parameters were lowest among the clinical isolates. Further, the responses induced by LAM-6 from modern lineage 4 were at a moderate level, similar to the laboratory strain H37Rv which also belongs to lineage 4. Thus, these profiles were specific to their respective lineages and/or genotypes and independent of their drug resistance status. Further, a positive correlation, among TNF-α, IL-1β, IL-6 and IL-12 induced in infected THP-1 cells was demonstrated. In addition, induction of all pro-inflammatory cytokines correlated well with the host cell apoptosis. A positive correlation was observed between phagocytic index in the category of '>10 bacilli/cell' and induction of apoptosis, only for virulent strains, indicating that initial accumulation of MTB strains inside the host cell may be an important determining factor for different innate responses.

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The apoptosis of infected THP-1 cells.(A) Flowcytometric profile of Annexin V-FITC labeled THP-1 cells infected with various MTB strains on 5th day of infection from a representative experiment of the three independent experiments. RN1 indicates the percentage of Annexin-V positive cells. (B) The bar diagram represents the percentage of infected THP-1 cells stained with AnnexinV after 5 and 6 days of infection. (C) Apoptosis was also determined using the Cell Death Detection ELISA (Roche Applied Science) after five days of infection. The bar diagram represents the extent of nucleosome fragmentation in infected THP-1 cells. (D) Expression of degraded PARP and β-Actin was monitored in THP-1 cells after 48 h of infection with five different MTB strains by Western Blotting. In this representative picture, the lanes were aligned from original picture.
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pone-0062966-g004: The apoptosis of infected THP-1 cells.(A) Flowcytometric profile of Annexin V-FITC labeled THP-1 cells infected with various MTB strains on 5th day of infection from a representative experiment of the three independent experiments. RN1 indicates the percentage of Annexin-V positive cells. (B) The bar diagram represents the percentage of infected THP-1 cells stained with AnnexinV after 5 and 6 days of infection. (C) Apoptosis was also determined using the Cell Death Detection ELISA (Roche Applied Science) after five days of infection. The bar diagram represents the extent of nucleosome fragmentation in infected THP-1 cells. (D) Expression of degraded PARP and β-Actin was monitored in THP-1 cells after 48 h of infection with five different MTB strains by Western Blotting. In this representative picture, the lanes were aligned from original picture.

Mentions: Significant apoptosis was observed after 5 days of infection in THP-1 cells as seen from flowcytometric data in Fig. 4A. The extent of apoptosis induced by different strains in THP-1 cells assessed by Annexin V labeling (Fig. 4B) and apoptosis ELISA (Fig. 4C), was compared after five and six days of infection. Among the five strains, H37Ra and EAI-5 induced significantly higher apoptosis compared to that in uninfected cells and cells infected by Beijing strain. Further, the apoptotic response to LAM-6 was similar to that observed for H37Rv. The degradation profile of pro-apoptotic protein, PARP in uninfected and infected THP-1 cells after 48 hrs of infection is depicted in Figure 4D and also corroborates the extent of apoptosis.


Drug resistant clinical isolates of Mycobacterium tuberculosis from different genotypes exhibit differential host responses in THP-1 cells.

Chakraborty P, Kulkarni S, Rajan R, Sainis K - PLoS ONE (2013)

The apoptosis of infected THP-1 cells.(A) Flowcytometric profile of Annexin V-FITC labeled THP-1 cells infected with various MTB strains on 5th day of infection from a representative experiment of the three independent experiments. RN1 indicates the percentage of Annexin-V positive cells. (B) The bar diagram represents the percentage of infected THP-1 cells stained with AnnexinV after 5 and 6 days of infection. (C) Apoptosis was also determined using the Cell Death Detection ELISA (Roche Applied Science) after five days of infection. The bar diagram represents the extent of nucleosome fragmentation in infected THP-1 cells. (D) Expression of degraded PARP and β-Actin was monitored in THP-1 cells after 48 h of infection with five different MTB strains by Western Blotting. In this representative picture, the lanes were aligned from original picture.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3646887&req=5

pone-0062966-g004: The apoptosis of infected THP-1 cells.(A) Flowcytometric profile of Annexin V-FITC labeled THP-1 cells infected with various MTB strains on 5th day of infection from a representative experiment of the three independent experiments. RN1 indicates the percentage of Annexin-V positive cells. (B) The bar diagram represents the percentage of infected THP-1 cells stained with AnnexinV after 5 and 6 days of infection. (C) Apoptosis was also determined using the Cell Death Detection ELISA (Roche Applied Science) after five days of infection. The bar diagram represents the extent of nucleosome fragmentation in infected THP-1 cells. (D) Expression of degraded PARP and β-Actin was monitored in THP-1 cells after 48 h of infection with five different MTB strains by Western Blotting. In this representative picture, the lanes were aligned from original picture.
Mentions: Significant apoptosis was observed after 5 days of infection in THP-1 cells as seen from flowcytometric data in Fig. 4A. The extent of apoptosis induced by different strains in THP-1 cells assessed by Annexin V labeling (Fig. 4B) and apoptosis ELISA (Fig. 4C), was compared after five and six days of infection. Among the five strains, H37Ra and EAI-5 induced significantly higher apoptosis compared to that in uninfected cells and cells infected by Beijing strain. Further, the apoptotic response to LAM-6 was similar to that observed for H37Rv. The degradation profile of pro-apoptotic protein, PARP in uninfected and infected THP-1 cells after 48 hrs of infection is depicted in Figure 4D and also corroborates the extent of apoptosis.

Bottom Line: EAI-5 strain from ancient lineage 1, induced higher proinflammatory responses, higher apoptosis and moderate intracellular growth compared to other strains, in contrast, for Beijing strain of modern lineage 2, all three parameters were lowest among the clinical isolates.Thus, these profiles were specific to their respective lineages and/or genotypes and independent of their drug resistance status.Further, a positive correlation, among TNF-α, IL-1β, IL-6 and IL-12 induced in infected THP-1 cells was demonstrated.

View Article: PubMed Central - PubMed

Affiliation: Radiation Medicine Centre, Bio-Medical Group, Bhabha Atomic Research Centre, Mumbai, India.

ABSTRACT
Mycobacterium tuberculosis (MTB) persistently infects and survives within the host macrophages. Substantial genotypic variation exists among MTB strains which correlate with their interactions with the host. The present study was designed to establish a correlation, if any, between infection and induction of innate immune response by genetically diverse drug resistant MTB isolates from India. For this purpose, three clinical isolates from ancient and modern lineages, along with H37Ra and H37Rv were evaluated for intracellular growth, phagocytic index, induction of proinflammatory cytokines and apoptosis following infection in THP-1 cell line. A wide variation in the induction of cytokines was revealed subsequent to infection with different strains. EAI-5 strain from ancient lineage 1, induced higher proinflammatory responses, higher apoptosis and moderate intracellular growth compared to other strains, in contrast, for Beijing strain of modern lineage 2, all three parameters were lowest among the clinical isolates. Further, the responses induced by LAM-6 from modern lineage 4 were at a moderate level, similar to the laboratory strain H37Rv which also belongs to lineage 4. Thus, these profiles were specific to their respective lineages and/or genotypes and independent of their drug resistance status. Further, a positive correlation, among TNF-α, IL-1β, IL-6 and IL-12 induced in infected THP-1 cells was demonstrated. In addition, induction of all pro-inflammatory cytokines correlated well with the host cell apoptosis. A positive correlation was observed between phagocytic index in the category of '>10 bacilli/cell' and induction of apoptosis, only for virulent strains, indicating that initial accumulation of MTB strains inside the host cell may be an important determining factor for different innate responses.

Show MeSH
Related in: MedlinePlus