Virulence factor RNA transcript expression in the Leishmania Viannia subgenus: influence of species, isolate source, and Leishmania RNA virus-1
Bibliographic record
Abstract
Leishmania RNA virus-1 (LRV1) is a double-stranded RNA virus identified in 20–25% of Viannia —species endemic to Latin America, and is believed to accelerate cutaneous to mucosal leishmaniasis over time. Our objective was to quantify known virulence factor (VF) RNA transcript expression according to LRV1 status, causative species, and isolate source. Eight cultured isolates of Leishmania were used, four of which were LRV1-positive ( Leishmania Viannia braziliensis [ n = 1], L . ( V .) guyanensis [ n = 1], L . ( V .) panamensis [ n = 2]), and four were LRV1-negative ( L . ( V .) panamensis [ n = 3], L . ( V .) braziliensis [ n = 1]). Promastigotes were inoculated into macrophage cultures, and harvested at 24 and 48 h. RNA transcript expression of hsp23 , hsp70 , hsp90 , hsp100 , mpi , cpb , and gp63 were quantified by qPCR. RNA transcript expression of hsp100 ( p = 0.012), cpb ( p = 0.016), and mpi ( p = 0.022) showed significant increases from baseline pure culture expression to 24- and 48-h post-macrophage infection, whereas hsp70 ( p = 0.004) was significantly decreased. A trend toward increased transcript expression of hsp100 at baseline in isolates of L . ( V .) panamensis was noted. Pooled VF RNA transcript expression by L . ( V .) panamensis isolates was lower than that of L . ( V .) braziliensis and L . ( V .) guyananesis at 24 h ( p = 0.03). VF RNA transcript expression did not differ by LRV1 status, or source of cultured isolate at baseline, 24, or 48 h; however, a trend toward increased VF RNA transcript expression of 2.71- and 1.93-fold change of mpi ( p = 0.11) and hsp90 (p = 0.11), respectively, in LRV1 negative isolates was noted. Similarly, a trend toward lower levels of overall VF RNA transcript expression in clinical isolates (1.15-fold change) compared to ATCC® strains at 24 h was noted ( p = 0.07). Our findings suggest that known VF RNA transcript expression may be affected by the process of macrophage infection. We were unable to demonstrate definitively that LRV-1 presence affected VF RNA transcript expression in the species and isolates studied. L . ( V .) guyanensis and L . ( V .) braziliensis demonstrated higher pooled VF RNA transcript expression than L . ( V .) panamensis ; however, further analyses of protein expression to corroborate this finding are warranted.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.001 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.001 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.001 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".