Expression of Trichinella spiralis serpin Tsp_01570 in Pichia pastoris: a first insight of its biomodulatory activity.

Serpins represent one of the most diverse families of serine protease inhibitors. Despite their complexity, they are virtually found in all organisms and play an important role in homeostasis processes such as blood coagulation, inflammation, fibrinolysis, immune responses, chromatin condensation, tumor suppression, and apoptosis. There has recently been particular interest in studying serpin functions in infection and inflammation, especially since more serpins from parasites have been identified and characterized. Among helminths, Trichinella spiralis is one of the few parasites with an extremely strong ability to induce host immune suppression. Previous studies show that serpins are present in Trichinella and are expressed differentially at different parasite stages. More interesting, there is evidence of a recombinant serpin from Trichinella pseudospiralis that alters macrophage polarization in vitro. This finding could be relevant to comprehend the modulation process of the immune response. We expressed Tsp_01570, a putative serpin gene from Trichinella spiralis, in the eukaryotic system Pichia pastoris SMD1168H and evaluated its presence at different parasite stages, finding the serine protease inhibitor in the crude extract of adult worms. The effect of recombinant serpin on THP-1 cells was tested by quantification of IL-12p40, TNF-α, IL-4, and IL-10 cytokines released by ELISA. We also evaluated the expression of the M1 markers, CCR7 and CD86, and the M2 markers, CD163 and CD206, by immunofluorescence staining. This study represents the first insight in elucidating the importance of serpin Tsp_01570 as a potential molecular modulator.

RDRioMycobacterium tuberculosis strains associated with isoniazid resistance in Northern Mexico.

BACKGROUND: It has been established that the genomic background of Mycobacterium tuberculosis may influence disease progression, in particular for the Beijing family and the Latin American and Mediterranean (LAM)/RDRio strains. The purpose of this study was to evaluate the prevalence of the LAM/RDRio genotype in cases of tuberculosis from Mexico and their drug susceptibility profile. METHODS: Two hundred eighteen M. tuberculosis isolates were screened by 43-spacer spoligotyping. The LAM/RDRio genotype was confirmed by multiplex PCR, and the drug susceptibility testing was carried out in solid Löwenstein-Jensen media. RESULTS: Among the LAM strains identified, 24 (63.1%) were confirmed as M. tuberculosis RDRio. All RDRio strains shared the RD174 deletion, that was associated with isoniazid resistance (p=0.0264). CONCLUSIONS: We documented for the first time the isolation of the LAM/RDRio genotype in pulmonary cases of tuberculosis in Mexico, and we found resistance to the first-line anti-tuberculosis drug isoniazid in these strains.

RDRioMycobacterium tuberculosis strains associated with isoniazid resistance in Northern Mexico.

BACKGROUND: It has been established that the genomic background of Mycobacterium tuberculosis may influence disease progression, in particular for the Beijing family and the Latin American and Mediterranean (LAM)/RDRio strains. The purpose of this study was to evaluate the prevalence of the LAM/RDRio genotype in cases of tuberculosis from Mexico and their drug susceptibility profile. METHODS: Two hundred eighteen M. tuberculosis isolates were screened by 43-spacer spoligotyping. The LAM/RDRio genotype was confirmed by multiplex PCR, and the drug susceptibility testing was carried out in solid Löwenstein-Jensen media. RESULTS: Among the LAM strains identified, 24 (63.1%) were confirmed as M. tuberculosis RDRio. All RDRio strains shared the RD174 deletion, that was associated with isoniazid resistance (p=0.0264). CONCLUSIONS: We documented for the first time the isolation of the LAM/RDRio genotype in pulmonary cases of tuberculosis in Mexico, and we found resistance to the first-line anti-tuberculosis drug isoniazid in these strains.