TLR1 Polymorphism Frequency in Different Leprosy Types.

1<p>P value for comparison of allele frequencies.</p>2<p>P value for comparison of genotype frequencies.</p>3<p><b><i>A</i></b> denotes common allele, <b><i>a</i></b> denotes minor allele.</p>4<p>Tuberculoid includes TT and BT. Lepromatous includes LL, BL, and BB.</p

Baseline characteristics of leprosy subjects.

1<p>Percentages of individuals in some categories do not sum to 100 due to missing data. Not shown among all leprosy are 8 subjects with peripheral neuropathy (PN), and one with leprosy of indeterminate type (IN).</p>2<p>The four most frequent ethnic groups (from > 8 ethnic groups) are tabulated.</p>3<p>P represents exact <i>P</i> values for overall distribution of gender and age groups within the different ethnic groups and for the overall distribution of leprosy type and immune reactions among the 4 different ethnic groups.</p

IL-β and TNF-α production in human mononuclear cells following stimulation with <i>M. leprae</i>.

<p>Peripheral blood mononuclear cells were stimulated for 18 hours and supernatants were assayed for cytokine production by luminex multiplex bead assay. PBMCs were obtained from 11 individuals with the genotype 1805TT or 1805TG (TT/TG: dark circles) and 10 individuals with the 1805GG genotypes (GG: open circles). PBMCs stimulated with media, PAM2 at 75 ng/mL (P2), PAM3 at 75 ng/mL (P3), or LPS at 10 ng/ml (LPS), whole irradiated ML at 20 µg/ml (ML) or MLcw at 10 µg/ml (MLcw). IL-1β production following stimulation is shown in (A), and TNF-α production in (B). The mean level and standard error of the mean are depicted and were derived from averaging the responses of individual within each genotype group. The median level is depicted by a bar; *p≤0.05, **p≤0.001 by Mann–Whitney U-test.</p

IL-6 production by human primary cells following stimulation with <i>M. leprae.</i>

<p>Peripheral blood mononuclear cells were stimulated for 18 hours and supernatants were assayed for cytokine production by ELISA. PBMCs were derived from whole blood taken from 15 individuals with the genotype 1805TTor 1805TG (TT/TG: dark circles) and 13 individuals with the 1805GG genotypes (GG: open circles). (A): PBMCs stimulated in triplicate with media, PAM2 at 75 ng/ml (P2 75), PAM3 at 75 ng/ml (P3 75), or LPS at 10 ng/ml (LPS 10). (B): PBMCs stimulated in triplicate with whole irradiated ML at 20 or 100 µg/ml (ML20 or ML100) or MLcw at 2 or 10 µg/ml (MLcw 2 or MLcw10). The mean level and standard error of the mean are depicted and were derived from averaging the responses of individuals stimulated in triplicate. The median level is depicted by a bar. *P≤0.01, **P≤0.001 by Mann–Whitney U-test.</p

NF-κB activity in response to <i>M. leprae</i> is diminished in the 1805T variant.

<p>HEK293 cells were transfected with an NF-κB luciferase reporter, a <i>Renilla</i> luciferase construct to control for transfection efficiency (pRL-TK), and CD14. Additional transfectants varied by condition and included: empty plasmid vector (EV, clear), TLR2 alone (T2, gray), or TLR2 with one of two TLR1 constructs, 1805T (black) or 1805G (stippled). Luciferase activity represents basal (media stimulation) or stimulated activity of transfected cells. Mean values (+/−standard deviation) are depicted for two representative experiments, each performed in triplicate. RLU, relative luciferase units; P3 300, PAM3 at a dose of 300 ng/ml; M2 100, Malp-2, at 100 ng/ml; ML, whole, irradiated ML at 5, 50, or 250 µg/ml; MLcw, <i>M. leprae</i> cell wall at a dose of 1 or 10 µg/ml. * = P≤0.01, by Student's t-test when comparing the 1805T and 1805G variants (both with T2); # = P≤0.01, by Student's t-test when comparing T2+1805T and T2; ˆ = P≤0.01, by Student's t-test when comparing T2+1805G and T2.</p

Frequency of TLR1 haplotypes in reversal reaction.

1<p>Order for 6 SNP haplotypes, left to right: Trs5743563C, Ars5743565G, Trs5743592C, Trs5743595C, G743A, T1805G.</p>2<p>Order for 2 SNP haplotypes: Trs5743592C, T1805G.</p>3<p>P value represents comparison of a given haplotype with the reference haplotype TATTAT.</p

TLR1 Polymorphism Frequency in Reversal Reaction.

1<p>P value for comparison of allele frequencies by Chi-square.</p>2<p>P value for comparison of genotype frequencies calculated by Chi-square unless otherwise indicated.</p>3<p><b><i>A</i></b> denotes common allele, <b><i>a</i></b> denotes minor allele.</p>4<p>P value calculated by Fisher's Exact test due to small cell number.</p

Number of positive responders and mean induration compared to ECs.

a<p>To allow calculations, the EC percent positive has been changed to 1.0.</p

Phase II, Stage A/B: Frequency of induration size (EC relative to PPD).

a<p>Subjects responding to MLSA-LAM low dose also responded to MLSA-LAM high dose.</p>b<p>The number of responders against <i>M. leprae</i> skin test antigens is derived from the high dose of both antigens.</p

Dot plot of induration measurements.

<p>Induration results are provided across five subject groups, including BL/LL leprosy patients (<i>n</i> = 19 in low and <i>n</i> = 20 in high dose group), BT/TT leprosy patients (<i>n</i> = 20), HC (<i>n</i> = 20), TB (<i>n</i> = 20), and ECs (<i>n</i> = 50). Low and high dose groups were combined to show PPD responses: BL/LL leprosy patients (<i>n</i> = 39) and all other groups (<i>n</i> = 40). Mean and standard deviation are shown.</p